[Observation of visual quality after implantation of a rotational asymmetric refractive intraocular lens with a low addition region using the micromonovision design].

Objective: To investigate the effects of the application of a low addition refractive multifocal intraocular lens (IOL) using the micromonovision design in the non-dominant eye with different degrees of preset myopia on the visual acuity, visual function and visual quality after bilateral cataract surgery. Methods: In this randomized controlled trial, patients who were proposed to undergo bilateral phacoemulsification combined with rotational asymmetric refractive IOL (MF15 IOL) implantation at the First Affiliated Hospital of Zhengzhou University between September 2020 and August 2022 were included. All patients were divided into three groups using the random number method. The target refraction of the IOL in the dominant eye was 0.00 D. Non-dominant eyes were given different preoperative IOL reserve refractions, with the reserved near additional degree>0.20 D and≤0.40 D as the low addition,>0.40 D and≤0.60 D as the medium addition, and>0.60 D and≤0.80 D as the high addition. We compared uncorrected distance visual acuity (UDVA), uncorrected intermediate visual acuity (UIVA) and uncorrected near visual acuity (UNVA) of monocular and binocular eyes at 1 day, 1 month and 3 months postoperatively in the 3 groups of patients. Furthermore, the contrast sensitivity, stereopsis, defocus curves and visual quality questionnaire results of binocular eyes were compared at 3 months postoperatively. The statistical methods mainly used were chi-square test, two-factor repeated measures ANOVA, one-way ANOVA, LSD test, Kruskal-Wallis test, and paired t-test. Results: A total of 110 patients (220 eyes) were enrolled in the study, including 48 males and 62 females, with an average age of (59.74±9.38) years. There were 40 patients (80 eyes) in the low additional degree group, 37 patients (74 eyes) in the medium additional degree group, and 33 patients (66 eyes) in the high additional degree group. The differences in distance, intermediate and near visual acuity of the dominant eyes among the three groups were not statistically significant at different measurement timepoints postoperatively (P>0.05). The differences in intermediate and near visual acuity of the non-dominant eyes were also not statistically significant (P>0.05) among the three groups. In contrast, at 3 months, the UDVA of the non-dominant eyes in the low additional degree group (0.04±0.06) and medium additional degree group (0.04±0.07) was significantly higher than that in the high additional degree group (0.08±0.09) (F=4.776, P=0.011, bias η2=0.086). There was no statistically significant difference in binocular uncorrected distance, intermediate and near visual acuity among the three groups at different postoperative timepoints (P>0.05). The binocular UDVA, UIVA and UNVA (logMAR visual acuity) at 3 months postoperatively were -0.04±0.04, 0.03±0.08, 0.10±0.13 in the low addition group, -0.01±0.05, -0.02±0.06, 0.09±0.10 in the medium addition group, and 0.02±0.07, 0.01±0.09, 0.16±0.11 in the high addition group. At 3 months postoperatively, the binocular contrast sensitivity of the low additional degree group was significantly higher than that of the high additional degree group (P<0.05), except that there was no significant difference at the spatial frequency of 6 cycles per degree in the absence of glare (P>0.05). The binocular contrast sensitivity of the medium additional degree group was significantly higher than that of the high additional degree group at the spatial frequencies of 6 and 18 cycles per degree in the glare condition (P<0.05). The difference in the binocular contrast sensitivity between the low and medium additional degree groups did not reach statistical significance (P>0.05). The peak of the binocular defocus curve in the three groups was significantly wider than that in the monocular eyes, and the decline trend was more gentle, with no trough in the middle, and the visual acuity could be maintained above 0.2 (logMAR visual acuity) in the 0.00 D to -3.00 D defocus range. There was no significant difference in the postoperative near stereopsis results among the three groups (P>0.05), with the percentage of near stereopsis sharpness≤60″ reaching 90.00% (36/40), 89.19% (33/37) and 78.79% (26/33), respectively. The proportions of VF-14 scores≥90 in the postoperative questionnaire were 90% (36/40), 91.89% (34/37) and 81.82% (27/33) for the low, medium and high additional degree groups, respectively. The differences in the probability of photic phenomena and spectacles-independent rate were not statistically significant (P>0.05). Conclusion: The use of micromonovision design for bilateral implantation of a rotational asymmetric refractive MF15 IOL, with the non-dominant eye reserved for different near additional degrees, can enable cataract patients to have significantly improved binocular full-range vision, visual function and visual quality. When the degree of reserved near additions in the non-dominant eye preoperatively is>0.20 D and≤0.60 D, it can ensure sufficient binocular UDVA, UIVA and UNVA after surgery, and meanwhile help to obtain superior contrast sensitivity and stereopsis, as well as a satisfactory spectacles-independent rate and low incidence of photic phenomena.

[Application effects of 3D visualization reconstruction technique in pheochromocytoma/ paraganglioma surgery].

To investigate the value of 3D visualization reconstruction technology in pheochromocytoma/paraganglioma surgery.The clinical data of 87 patients with pheochromocytoma/paraganglioma admitted to the Department of Urology of Peking Union Medical College Hospital between January 2019 and December 2022 were retrospectively analyzed, and 3D visualization model reconstruction was performed preoperatively in 47 patients [Group A:males was 24 cases,the age M(Q1, Q3)42.00(30.00, 54.00)]. while the remaining 40 patients [Group B: males was 23 cases,the age M(Q1, Q3) 44.00(30.25, 53.75)] was not. The maximum tumor diameter, operation time, intraoperative bleeding, drain retention time and postoperative hospital stay were compared between the two groups. Surgery was successfully completed in both groups. 37 (78.7%) patients in group A underwent laparoscopic surgery, 7 (14.9%) patients underwent open surgery, and 3 (6.4%) patients underwent laparoscopic-to-open surgery. Thirty-one (77.5%) patients in group B underwent laparoscopic surgery, 5 (12.5%) patients underwent open surgery, and 4 (10.0%) patients underwent laparoscopic to open surgery. There was a difference in the maximum diameter of the tumor between the two groups [(6.09±3.02) cm vs (5.32±1.76) cm, P<0.05], the retention time of the drainage tube was significantly shorter in group A compared with group B [(3.20±1.38) d vs (4.02±1.98) d, P<0.05], and the length of the hospital stay after surgery was significantly shorter [(5.75±2.12) d vs (6.49±3.37) d, P<0.05]. Comparison of operation time and intraoperative bleeding between the two groups showed no statistically significant difference (P>0.05).Two cases of postoperative anemia and one case of pulmonary atelectasis in group B patients improved before discharge. Conclusion when the tumor diameter is>6 cm or has a close relationship with the surrounding organs and blood vessels, the use of 3D visual reconstruction technology can formulate and implement a more accurate and safe surgical plan, shorten the retention time of the drainage tube and postoperative hospitalization time, which is conducive to the patient's postoperative recovery and reduce postoperative complications.

[Clinical observation of spontaneous brain activity in children with congenital cortical cataract amblyopia].

Objective: To investigate the characteristics of spontaneous brain activity in children with congenital cortical cataract amblyopia. Methods: A cross-sectional study was conducted. Twenty cases of unilateral congenital cortical cataract amblyopia (unilateral amblyopia group) and 14 cases of bilateral congenital cortical cataract amblyopia (bilateral amblyopia group) were enrolled from January 2022 to December 2022 at the First Affiliated Hospital of Zhengzhou University. Seventeen age and gender matched children with normal visual acuity were recruited as the healthy control group. Resting-state functional MRI (fMRI) was performed on all participants, and the amplitude of low-frequency fluctuations (ALFF) technique was used to analyze their spontaneous brain activities. The original ALFF value of each voxel was divided by the average ALFF value of the whole brain to obtain the standardized ALFF value (referred to as ALFF value), which reflected the intensity of spontaneous brain activity in different brain regions. General demographic data were compared using one-way analysis of variance, Kruskal-Wallis test, and chi-square test. Comparison of ALFF values was conducted using one-way analysis of variance. Results: There were no significant differences in age, gender, distribution of amblyopic eye or non-dominant eye, and degree of refractive error among the three groups (all P>0.05). Compared to the healthy control group, the unilateral amblyopia group showed higher ALFF values in the right posterior lobe of the cerebellum (67 voxels, t=3.48) and left posterior lobe of the cerebellum (71 voxels, t=4.09), and lower ALFF values in the right postcentral gyrus (91 voxels, t=-3.91), right inferior parietal lobule (73 voxels, t=-4.88), right inferior frontal gyrus (78 voxels, t=-4.09), left inferior parietal lobule (556 voxels, t=-4.82), and left inferior frontal gyrus (122 voxels, t=-4.27) (all P<0.01). The bilateral amblyopia group showed higher ALFF values in the right insula (60 voxels, t=3.54), right Rolandic operculum (69 voxels, t=3.73), right posterior lobe of the cerebellum (54 voxels, t=3.43), and left posterior lobe of the cerebellum (143 voxels, t=3.69), and lower ALFF values in the left inferior frontal gyrus (99 voxels, t=-4.39), left postcentral gyrus (231 voxels, t=-4.28), and right inferior parietal lobule (54 voxels, t=-3.77) (all P<0.01). Compared to the unilateral amblyopia group, the bilateral amblyopia group showed higher ALFF values in the left middle frontal gyrus (52 voxels, t=3.15, P=0.029), left posterior lobe of the cerebellum (77 voxels, t=3.39, P=0.001), and right Rolandic operculum (53 voxels, t=3.59, P=0.007). Conclusion: Children with congenital cortical cataract amblyopia exhibit altered spontaneous brain activity in multiple brain regions, and there are differences in spontaneous brain activity changes between unilateral and bilateral amblyopia.

Phenotype and genetic characteristics in 20 Chinese patients with 46,XY disorders of sex development.

PURPOSE: 46,XY disorders of sex development (DSD) is the most complicated and common type of DSD. To date, more than 30 genes have been identified associated with 46,XY DSD. However, the mutation spectrum of 46,XY DSD is incomplete owing to the high genetic and clinical heterogeneity. This study aims to provide clinical and mutational characteristics of 18 Chinese patients with 46,XY DSD. METHODS: A total of 20 unrelated individuals with 46,XY DSD were recruited. Whole-exome sequencing (WES) or custom-panel sequencing combined Sanger sequencing were performed to detect the pathogenic mutations. The pathogenicity of the variant was assessed according to the American College of Medical Genetics and Genomics (ACMG) guidance and technical standards recommended by the ACMG and the Clinical Genome Resource (ClinGen). RESULTS: Six patients harbored NR5A1 mutations; two patients harbored NR0B1 mutations; six patients harbored SRD5A2 mutations; six patients harbored AR mutations. Six novel genetic variants were identified involved in three genes (NR5A1, NR0B1, and AR). CONCLUSION: We determined the genetic etiology for all enrolled patients. Our study expanded the mutation spectrum of 46,XY DSD and provided diagnostic evidence for patients with the same mutation in the future.

[Visual quality after implantation of a rotational asymmetric refractive intraocular lens in patients with high myopia and cataract].

Objective: To evaluate the visual quality after phacoemulsification and implantation of a rotational asymmetric refractive intraocular lens in patients with high myopia and cataract. Methods: A retrospective cohort study. Sixty-two patients (92 eyes) of the First Affiliated Hospital of Zhengzhou University from June 2017 to December 2019 were evaluated after phacoemulsification and implantation of a rotational asymmetric refractive intraocular lens (LS-313 MF30). According to the axial length, the participants were allocated to either a control group or a high myopia group. Among the 33 patients (46 eyes) in the control group, the axial length was shorter than 26 mm; among the 29 patients (46 eyes) in the high myopia group, the axial length was more than 26 mm. The high myopia group was further divided into two subgroups. The super high myopia subgroup included 12 patients (18 eyes), and the axial length was ≥30 mm; the high myopia subgroup consisted of 17 patients (28 eyes), and the axial length was<30 mm and ≥26 mm. Uncorrected distance visual acuity, uncorrected intermediate visual acuity and uncorrected near visual acuity were recorded after surgery. The follow-up time was more than 6 months. At the end of the follow-up, patients' contrast sensitivity (CS), reading acuity, reading speed and visual quality questionnaire results were assessed. The main statistical methods were two-way repeated measures analysis of variance, independent sample t-test, one-way analysis of variance and Kruskal-Wallis test. Results: There were no significant differences in gender distribution, age, or follow-up time between the control and high myopia groups, the control and high myopia subgroup, or the super high myopia subgroup (all P>0.05). At the end of the follow-up, the uncorrected distance, intermediate and near visual acuity of the super high myopia subgroup was 0.07±0.11, 0.34±0.08 and 0.20±0.09, respectively. The difference in postoperative visual acuity was not statistically significant (all P>0.05). The CS and CS with glare of the high myopia group (3 cpd: 1.48±0.18, 1.42±0.16; 6 cpd: 1.75±0.18, 1.76±0.15; 12 cpd: 1.44±0.24, 1.43±0.19; 18 cpd: 0.90±0.23, 0.85±0.20) were significantly different from the control group (3 cpd: 1.66±0.18, 1.62±0.16; 6 cpd: 1.88±0.14, 1.85±0.11; 12 cpd: 1.54±0.18, 1.53±0.14; 18 cpd: 1.06±0.18, 1.01±0.15) (P<0.05), except at 12 cpd (P=0.05). The CS and CS with glare of the super high myopia subgroup (3 cpd: 1.39±0.21, 1.31±0.13; 6 cpd: 1.66±0.16, 1.64±0.12; 12 cpd: 1.32±0.23, 1.31±0.18; 18 cpd: 0.75±0.16, 0.73±0.15) were worst (all P<0.05). A significant difference between the high myopia subgroup and the control group was only found at 3 cpd (1.53±0.13 vs. 1.66±0.18, 1.50±0.13 vs. 1.62±0.16; both P<0.05). The reading acuity and reading speed of the super high myopia subgroup were lower than the high myopia subgroup and the control group, while the differences were not statistically significant (all P>0.05). The questionnaire survey showed that there was no significant difference in the proportion of glare and halo between the two subgroups and the control group (both P>0.05). No patients reported dissatisfaction. The rate of glasses independents was 11/12 in the super high myopia subgroup, 15/17 in the high myopia subgroup and 31/33 in the control group, with no significant difference (P>0.05). Conclusions: The rotational asymmetric refractive intraocular lens is suitable for patients with high myopia and cataract, and has good far and near vision after operation. However, it could result in relatively low CS for super high myopia patients, so rigorous preoperative examination must be done. (Chin J Ophthalmol, 2021, 57: 358-365).

[Effect of alprostadil in early wound healing of rats with deep partial-thickness scald].

Objective: To explore the effect of alprostadil in early wound healing of rats with deep partial-thickness scald. Methods: Ninety specific pathogen free healthy Sprague-Dawley rats (half males and half females) were divided into sham scald group, simple scald group, and alprostadil group using the random number table with 30 rats in each group. Rats in sham scald group were sham injured, and rats in the other 2 groups were inflicted with deep partial-thickness scald of 30% total body surface area on the back. Immediately after scald, rats in the 3 groups received anti-shock treatment. Two hours post scald, rats in sham scald group and simple scald group were injected intraperitoneally with 1 mL normal saline, and rats in alprostadil group were injected intraperitoneally with 1 mL alprostadil injection, once a day and continued for 14 days. On post scald day (PSD) 3, 7, and 14, 10 rats in each group were collected for observing the general wound conditions and the wound healing rates of rats in 2 scald groups were calculated, abdominal aortic blood of 2 mL from each rat were collected to detect serum thromboxane B2 by enzyme-linked immunosorbent assay, and wound tissue on the back was collected to examine pathomorphological change by hematoxylin-eosin staining and to detect wound microvessel density (MVD) by immunohistochemical staining. Data were statistically analyzed with analysis of variance of factorial design, t test, Kruskal-Wallis H test, Mann-Whitney U test, and Bonferroni correction. Results: (1) There was no scald wound in rats in sham scald group. On PSD 3, wounds of rats in simple scald group and alprostadil group formed dry eschar. On PSD 7 and 14, wound areas of rats in alprostadil group were significantly smaller than those of rats in simple scald group, and with less exudation. (2) On PSD 3, the wound healing rates of rats in simple scald group and alprostadil group were similar (t=1.167, P>0.05). On PSD 7 and 14, the wound healing rates of rats in alprostadil group were significantly higher than those in simple scald group (t=8.657, 33.050, P<0.01). (3) On PSD 3, 7, and 14, the levels of serum thromboxane B2 of rats in simple scald group and alprostadil group were (541±22), (607±47), (688±21), (326±25), (271±21), (135±27) pg/mL, significantly higher than (17±6), (16±4), (16±4) pg/mL of rats in sham scald group (t=72.977, 39.685, 102.076, 37.033, 37.253, 13.845, P<0.01). On PSD 3, 7, and 14, the levels of serum thromboxane B2 of rats in alprostadil group were significantly lower than those in simple scald group (t=20.637, 20.651, 51.680, P<0.01). (4) Normal epidermis and dermis were seen in rats in sham scald group. On PSD 3, a large number of necrotic tissue and inflammatory cells infiltration were seen in wounds of rats in simple scald group, while a little new epithelium formation and some inflammatory cells infiltration were seen in wounds of rats in alprostadil group. On PSD 7 and 14, the new epithelium of rats in alprostadil group was significantly thicker than that in simple scald group, and epidermis formed gradually in alprostadil group. (5) On PSD 3, 7, and 14, the wound MVD of rats in simple scald group and alprostadil group were significantly higher than those in sham scald group (Z=-3.780, -3.781, -3.780, -3.780, -3.781, -3.780, P<0.01). On PSD 3, the wound MVD of rats in simple scald group and alprostadil group were similar (Z=-1.965, P>0.05). On PSD 7 and 14, the wound MVD of rats in alprostadil group were significantly higher than those in simple scald group (Z=-3.780, -3.780, P<0.01). Conclusions: The early intervention with alprostadil can significantly improve microcirculation of deep partial-thickness scald wound, reduce inflammatory cell infiltration, promote the formation of new blood vessels, thus promoting wound healing.

MiR-212-5p inhibits the malignant behavior of clear cell renal cell carcinoma cells by targeting TBX15.

OBJECTIVE: To investigate the role of microRNA-212-5p (miR-212-5p) in clear cell renal cell carcinoma (ccRCC) and to explore the potential underlying mechanisms. MATERIALS AND METHODS: 32 pairs of ccRCC clinical samples were collected. Renal ccRCC cells (786-O) and embryonic kidney cells (293T) were cultured in vitro. The ability of cell proliferation was detected by 3-(4,5)-dimethylthiazol(-z-y1)-3,5-diphenyl tetrazolium bromide (MTT) assay. Transwell migration assay was used to detect the abilities of cell invasion and migration. The relative protein and mRNA expressions of miR-212-5p were detected by Western blot and quantitative Real-time polymerase chain reaction (qRT-PCR) analysis, respectively. Furthermore, bioinformatics online sites and luciferase reporter gene assay were performed to predict and verify the potential targets of miR-212-5p, respectively. RESULTS: The expression level of miR-212-5p in ccRCC tissues and cell lines was significantly inhibited. Bioinformatics online sites and luciferase reporter gene assay confirmed that T-box transcription factor TBX15 (TBX15) was the potential target gene of miR-212-5p. In vitro experiments demonstrated that the proliferation, cell cycle, cell invasion and migration of ccRCC cells were obviously restricted after up-regulation of miR-212-5p. However, the above functional effects were significantly abolished in ccRCC cells after co-transfection with miR-212-5p mimics and LV-TBX15. CONCLUSIONS: MiR-212-5p acted as a tumor suppressor gene in ccRCC. Through targeting TBX15, miR-212-5p significantly inhibited the malignant behavior of ccRCC cells. Our findings revealed that miR-212-5p/TBX15 axis might be a potential therapeutic target for the treatment of ccRCC.

Effect of endostatin on proliferation, invasion and epithelial-mesenchymal transition of basal cell carcinoma cell A431.

OBJECTIVE: To investigate the effect of endostatin on the proliferation, invasion and epithelial-mesenchymal transition (EMT) of human basal cell carcinoma (BCC) cells (A431). PATIENTS AND METHODS: CCK-8 assay and transwell chamber assay were performed to detect cell proliferation and invasion abilities, respectively. Western blot was performed for the detection of the expressions of EMT-related proteins levels. The therapeutic effect of endostatin on tumor formation was tested using a mouse xenograft model. RESULTS: After endostatin treatment, transwell assay showed that the number of invasive cells in the observation group and control group were (38.25±8.13) and (98.25±9.14), respectively; the relative expression level of E-cadherin protein in the observation group was (0.34±0.03), which was significantly higher than that in the control group (0.14±0.01); the relative expression levels of N-cadherin protein in the observation group was (0.18±0.05), which was significantly lower than that in the control group (0.43±0.03), (all p<0.05). CONCLUSIONS: The expression levels of Vimentin and Fibronectin proteins were significantly lower, while the expression levels of α-smooth muscle Actin (α-SMA) were significantly higher in the observation group than those in the control group. Treatment with endostatin significantly inhibited tumor growth in the mouse xenograft model. Therefore, endostatin can inhibit the proliferation, invasion and EMT in BCC.

[Effects of different time of cryoablation on the proliferation of airway traumatic granuloma and its mechanism in rabbits].

Objective: To investigate the effect of different cryoablation time on tracheal traumatic granulation formation and its mechanism. Methods: A total of 32 rabbits were randomly assigned into four groups (A-D). Group A underwent tracheotomy alone. Group B, C and D received intra-tracheal brush rubbing to establish airway granulation model. Group C and D underwent 30 s and 2-minute cryoablation respectively. Tracheal specimens of all groups were collected to examine pathological changes using HE staining. Levels of transforming growth factor beta 1 (TGF-ß(1)) and CD34 in tracheal granulation were evaluated using immunohistochemistry (IHC) and real-time quantitative reverse transcription polymerase chain reaction (RT-qCR). Results: Tracheal lumens of group A were smooth without granulation. While the growth of granulation and luminal stenosis were most severe in Group B, followed by Group D and C. Submucosa thickness of Group B was largest as compared with other groups (0.20±0.07, 0.77±0.28, 0.44±0.13 and 0.55±0.18 mm for Group A to D, respectively. P<0.05). And the submucosa layer of Group C was thinner than Group D (P<0.05). The expression and transcription levels of TGF-ß(1) of trachea were highest in Group B as detected by IHC and RT-qPCR (P<0.05), followed by Group D and C (IHC: 0.48±0.01 vs 0.43±0.01, P<0.05; RT-qPCR: 12.61±2.14 vs 2.38±0.10, P<0.05). Both protein and mRNA levels of CD34 were highest in Group B as detected by IHC and RT-qPCR (P<0.05). Tracheal mRNA levels of CD34 were more abundant in Group D than Group C (4.92±0.90 vs 2.09±0.10, P<0.05), while no significant difference was found between groups regarding protein levels measured by IHC. Conclusions: Cryoablation could alleviate the hyperplasia of tracheal traumatic granulation, possibly due to the inhibition of TGF-ß(1) and CD34 expression. The effect of 30 s cryoablation for tracheal traumatic granulation is better.

[Effect of alprostadil on wound healing of scalded rats and the mechanism].

Objective: To explore effect of alprostadil on wound healing of scalded rats and the mechanism. Methods: According to random number table method, forty-eight Sprague Dawley rats were divided into sham scald group, simple scald group, lithium chloride group, and alprostadil group, with 12 rats in each group. Rats in sham injury group were sham injured on the back, and rats in the other three groups were inflicted with 30% total body surface area deep partial thickness scald on the back.Immediately after scald, rats in sham scald group and simple scald group were injected with 1 mL saline through caudal vein, and rats in lithium chloride group and alprostadil group were injected respectively with 1 mL lithium chloride and alprostadil through caudal vein. Saline, lithium chloride, and alprostadil were injected once in a day and lasted for 14 days. General wound appearance and wound healing rate on post scald day (PSD) 7, 10, 14 were observed and calculated. Expressions of protein and mRNA of Wnt1 and ß-catenin on PSD 14 were detected. Data were processed with analysis of variance of factorial design, one-way analysis of variance, Student Newman Keuls q test, t test, and Bonferroni correction. Results: (1) On PSD 7, wounds of scalded rats in each group formed dry eschar and had little exudation. On PSD 10, wounds of rats in simple scald group were covered with eschar, with little exudation, and wounds of rats in lithium chloride group were covered with eschar, and partial wounds healed under the eschar. On PSD 10, partial eschar of rats in alprostadil group desquamated; partial wounds healed; newly burned skin was ruddy. On PSD 14, partial wounds of rats in simple scald group were healed under eschar with little exudation. On PSD 14, most of the eschar of rats in lithium chloride group were desquamated with patial wounds healed and little exudation. On PSD 14, wounds of rats in alprostadil group were basically healed with vigorously growing hair on the back. (2) On PSD 7, the wound healing rates of rats in simple scald group, lithium chloride group, and alprostadil group were close (F=0.41, P>0.05). On PSD 10 and 14, wound healing rate of rats in lithium chloride group and alprostadil group were significantly higher than that in simple scald group (q=5.73, 17.45, 26.30, 11.28, P<0.05), and wound healing rate of rats in alprostadil group was significantly higher than that in lithium chloride group (q=32.03, 28.73, P<0.05). (3) On PSD 14, the mRNA expressions of Wnt1 and ß-catenin of rats in lithium chloride group and alprostadil group were significantly higher than those in simple scald group (q=65.40, 19.16, 66.79, 18.41, P<0.05), and the mRNA expressions of Wnt1 and ß-catenin of rats in simple scald group was significantly higher than those in sham scald group (t=14.86, 4.46, P<0.05). (4) On PSD 14, the protein expressions of Wnt1 and ß-catenin of rats in lithium chloride group and alprostadil group were 0.98±0.05, 0.98±0.06, 0.97±0.06, and 1.00±0.06, which were significantly higher than 0.49±0.04 and 0.66±0.04 of rats in simple scald group (q=34.62, 22.38, 33.61, 23.47, P<0.05). On PSD 14, the protein expressions of Wnt1 and ß-catenin of rats in simple scald group was significantly higher than 0.29±0.03 and 0.31±0.03 of rats in sham scald group (q=14.73, 23.88, P<0.05). Conclusions: Alprostadil can accelerate wound healing through activating Wnt/ß-catenin signal pathway and upregulating the expressions of Wnt1 and ß-catenin.

[The effect of L-carnitine on the apoptosis of human lens epithelial cells through endoplasmic reticulum stress].

Objective: To investigate the effect of L-carnitine on the apoptosis of human lens epithelial cells through endoplasmic reticulum (ER) stress pathway. Methods: HLE-B3 cell lines were used to set up an oxidative stress model with H(2)O(2) treatment for 12 h, and lead to ER stress. Cells were divided into four groups: H(2)O(2) group, L-carnitine (100 µmol/L) with H(2)O(2) group, phosphate buffered saline (PBS) group and L-carnitine (100 µmol/L) group. Cell counting kit-8 was used to detect the cell viability under the treatment of different concentrations (200, 400, 600 and 800 µmol/L) of H(2)O(2). The apoptosis ratio of HLE-B3 treated by 400 µmol/L H(2)O(2) was tested by flow cytometry. When HLE-B3 was treated by 400 µmol/L H(2)O(2), the expression levels of cysteinyl aspartate specific proteinase 3 (caspase-3) gene and glucose-regulated protein 78 (GRP78) gene were measured by RT-PCR, and the expression levels of caspase-3 protein and GRP78 protein were assayed by Western blotting. Data from groups was analyzed by the one-way analysis of variance, and the LSD-t test was used for the comparison of groups. Results: Cell counting kit-8 assay showed that when H(2)O(2) concentration was 200, 400, 600 and 800 µmol/L, there was significant difference in the H(2)O(2) group(77.6%±0.8%,58.1%±3.1%,39.2%±1.5%,28.1%±2.2%), L-carnitine with H(2)O(2) group(83.3%±4.2%,74.5%±3.1%,46.4%±1.7%,32.4%±1.2%), PBS group(97.6%±2.1%,98.3%±0.2%,96.3%±2.2%,98.5%±1.1%) and L-carnitine group(98.5%±1.3%, 96.1%±2.1%, 98.1%±0.2%, 97.3%±1.4%) (all P<0.05). There was no significant difference between groups (PBS group compared to L-carnitine group, all P>0.05). When the concentration of H(2)O(2) was 400 µmol/L, the survival rate of the L-carnitine with H(2)O(2) group was higher than the H(2)O(2) group. The difference was statistically significant (t=18.14, P=0.020). With increasing of the H(2)O(2) concentration, cell necrosis was increased. The cell survival rate had no significant difference between the L-carnitine with H(2)O(2) group and H(2)O(2) group (both P>0.05). Flow cytometry results of the H(2)O(2) group, L-carnitine with H(2)O(2) group, PBS group and L-carnitine group were 31.4%±4.5%, 16.5%±2.8%, 2.1%±0.2% and 1.9%±1.8%, respectively (F=126.784, P=0.024) . The rate of apoptosis in the L-carnitine with H(2)O(2) group was lower than that in the H(2)O(2) group (t=24.67, P=0.013). There was no significant difference between the PBS group and L-carnitine group (P>0.05). The results of RT-PCR showed that the expression of caspase-3 mRNA in the L-carnitine with H(2)O(2) group was lower than the H(2)O(2) group (0.424±0.041 vs. 0.752±0.203), and the expression of GRP78 mRNA in the L-carnitine with H(2)O(2) group was lower than the H(2)O(2) group (0.521±0.223 vs. 0.821±0.103). The difference was statistically significant (caspase-3: t=27.92, P=0.018;GRP78: t=16.31, P=0.019). Western blotting showed that the protein expression of caspase-3 in the L-carnitine with H(2)O(2) group was lower than the H(2)O(2) group (0.712±0.212 vs. 1.126±0.251), and the GRP78 protein expression in the L-carnitine with H(2)O(2) group was lower than the H(2)O(2) group (0.512±0.012 vs. 0.735±0.051). The difference was statistically significant (caspase-3: t=15.43, P=0.010;GRP78: t=20.62, P=0.018). Conclusion: L-carnitine can reduce the apoptosis rate of HLE-B3 during oxidative stress through ER stress pathway. (Chin J Ophthalmol, 2018, 54: 363-368).

[The influence of EphA2 overexpression on proliferation and apoptosis of human lens epithelial cells exposed to high-concentration dexamethasone in vitro].

Objective: To construct lentiviral-mediated EphA2 overexpression vectors, transfect them into human lens epithelial cells (HLE-B3) in vitro, and investigate the effect of EphA2 gene overexpression on the proliferation and apoptosis of HLE-B3 exposed to high-concentration dexamethasone. Methods: Experimental Study. The pCDH-CMV- MCS-EF1-RFP plasmid was set up by the digestion of NOTⅠand XbaⅠ double restriction enzyme and ligation of CE ligase, and then the plasmid was transformed into DH10B cells. Seven clons were picked for enzymatic digestion and the clons with correct results were chosen for sequencing. The 293 T/17 cells were co-transfected with the pCDH-CMV-MCS-EF1-RFP-EphA2 and the packaging mixture by Lipofectamine 2000. At different multiplicities of infection (MOI=20, 50, 100, and 200) after 72-hour infection, we observed the expression of RFP and morphological changes of HLE-B3 by an inverted fluorescence microscope, and calculated the transfection efficiency through the flow cytometry. EphA2 protein expression was detected by Western blot. The following experiments were divided into four groups: normal control group (group A), EphA2 overexpression vector transfection group (group B), HLE-B3 cells exposed to dexamethasone group (group C) and EphA2 overexpression vector transfection HLE-B3 cells exposed to dexamethasone group (group D). Statistical analysis method was single factor or two factors variance analysis. Cell survival rate was detected by the Cell Counting Kit-8 assay. Cell apoptosis index was detected by Tunel. Results: Restriction enzyme digestion and sequencing indicated that EphA2 cDNA fragment was successfully inserted in the vector. The infection efficiency was up to 38.6%±3.9%, 49.2%±4.2%, 79.5%±5.5% and 80.2%±6.0% when the MOI was 20, 50, 100 and 200, respectively. There was statistically significant difference (F=2 600.8, P=0.001) among the four groups and between any two groups except between the MOI=100 group and MOI=200 group (P=2.507) . The relative quantity of EphA2 protein of the normal control group, empty vector transfection group and EphA2 gene overexpression vector transfection group was (0.561 2±0.031 7) , (0.559 7±0.012 8) and (3.032 0±0.041 9) , respectively. There was statistically significant difference (F=2 646.0, P=0.001) among the three groups and between any two groups except between the normal control group and empty vector transfection group (P=0.868) . The survival rate of groups A, B, C and D was 98.18%±1.85%, 122.01%±3.89%, 52.32%±1.99% and 76.18%±3.74%, respectively. There was statistically significant difference among the four groups (F=497.6, P=0.001) . The survival rate of group B was greater than group A (P=0.001) . The survival rate of group D was greater than group C (P=0.001) . Tunel results showed that the apoptosis index of groups A, B, C and D was 5.4%±1.5%, 5.0%±1.3%, 23.0%±3.9% and 14.4%±2.7%, respectively. There was statistically significant difference among the four groups (F=397.6, P=0.001) . The apoptosis index of group B was lower than group A, but there was no statistically significant difference between them (P=0.415) ; the apoptosis index of group D was lower than group C (P=0.018). Conclusions: The lentiviral vector carrying human EphA2 gene has been successfully constructed and efficiently expressed in HLE-B3 cells. EphA2 gene overexpression could increase the HLE-B3 cell survival rate and protect HLE-B3 cells from high-concentration dexamethasone-induced reduction of the cell survival rate. EphA2 gene overexpression could protect HLE-B3 cells from high-concentration dexamethasone-induced apoptosis, but it has no remarkable effect on apoptosis of HLE-B3 cells under physiological conditions. (Chin J Ophthalmol, 2018, 54: 125-132).

[Clinical observation of visual quality after the individual implantation of intraocular lens guided by corneal Q value].

Objective: To describe the feasibility of individual IOL implantation guided by corneal Q value and observe patients postoperative visual quality under different residual spherical aberration. Methods: Prospective study. One hundred and twenty cases (171 eyes)cataract patients in our hospital were selected for the individual implantation of intraocular lens guided by corneal Q value which obtained by Oberscan before operation. Based on spherical aberration calculated by corneal Q value, choose appropriate IOL personalitily to make postoperative whole eye surface aberration +0.1 µm (group of positive spherical aberration) or 0 µm (group of zero spherical aberration). To observe spherical aberration, the uncorrected visual acuity, best corrected visual acuity and contrast sensitivity (including no glare and glare) 1 month and 3 months after surgery. Dates were analyzed with one-way ANOVA and LSD method for multiple comparisons between groups. Results: Spherical aberration after operation: group of positive spherical aberration: (0.111±0.023)µm, group of zero spherical aberration: (0.020±0.019)µm, control group: (0.299±0.073)µm. At 1 months and 3 months, uncorrected visual acuity, and corrected visual acuity were not statistically different between groups (t=0.474, 1.607, P>0.05). Contrast sensitivity (including no glare and glare) 3 months after surgery display: at whole space frequency, the group of positive spherical aberration(reserved +0.1 µm spherical aberration) contrast sensitivity is better than that of the group of zero spherical aberration(reserved 0.0 µm spherical aberration) and the control group(F=32.885, 35.493, 19.969, 20.572,P<0.05). The group of zero spherical aberration is better than control group at space frequency of 3 and 6 c/d(F=6.506, 7.521, P<0.05). Conclusions: The individual implantation of introcular lens guided by corneal Q value is feasible. + 0.1 µm spherical aberration after surgery can achieve the best contrast sensitivity and stereo vision, and 0 spherical aberration after surgery can improve the postoperative contrast sensitivity and stereo vision than a traditional method, but its advantage mainly embodies in the middle and lower spatial frequency. (Chin J Ophthalmol, 2017, 53: 814-820).

[Long-term clinical observation of aspheric multifocal intraocular lens implantation].

Objective: To observe the clinical effect of long term visual quality after the implantation of the aspheric diffractive multifocal intraocular lens. Methods: This was a retrospective cohort study.One hundred and thirty cases of age-related cataract (170 eyes) after phacoemulsification cataract extraction combined with IOL implantation were collected from September of 2009 to January of 2011 in the First Affiliated Hospital of Zhengzhou University.There were 42 patients (57 eyes) with aspheric multifocal group, 43 patients (57 eyes) in the aspheric group and 45 patients (56 eyes) in the spherical group according to the different types of IOL implanted.At 1 year, 3 years and 5 years after operation, the following parameters were assessed: uncorrected and best corrected distance, medium and near vision acuity, contrast sensitivity, wavefront aberrations, modulation transfer functions (MTF), stereopsis, visual function and quality of life (VF/QOL) questionnaire survey. Results: At 1 year, 3 years and 5 years after operation, the uncorrected medium visual acuity in aspheric diffractive multifocal IOL group(0.30(0.10, 0.50), 0.30(0.10, 1.00), 0.30(0.10, 0.50)) was better than that of eyes in aspheric IOL group(0.40 (0.10, 0.0), 0.40 (0.20, 1.00), 0.40 (0.20, 0.50)) (Z(1)=-3.32,-1.73,-3.01, P(1)=0.00, 0.01, 0.00) and spherical IOL group (0.40(0.30, 1.00), 0.40(0.20, 1.00), 0.40(0.20, 1.00)) (Z(2)=-5.77,-3.19,-4.49, P(2)=0.00, 0.00, 0.00).And the near vision in aspheric diffractive multifocal IOL group(0.25(0.00, 1.00), 0.30(0.00, 1.00), 0.30(0.00, 1.00)) was also obviously better than that of eyes in aspheric IOL group (0.50(0.18, 1.00), 0.50(0.18, 1.00), 0.50(0.18, 1.00)) (Z(1)=-5.57,-5.37,-4.93, P(1)=0.00, 0.00, 0.00) and spherical IOL group(0.60(0.18, 1.00), 0.60(0.18, 1.00), 0.60(0.18, 1.00)) (Z(2)=-7.00,-6.91,-6.53, P(2)=0.00, 0.00, 0.00). At 5 years after operation, the mean higher-order aberration for 3.0mm and 5.0mm optical zone in aspheric diffractive multifocal IOL group (0.21(0.03, 0.46), 0.37(0.12, 2.01)) were significantly lower than that in spherical IOL group (0.43(0.10, 1.91), 0.46 (0.10, 1.91) ) (Z(2)=-4.81,-1.97, P(2)=0.00, 0.01).But there was no statistical difference between the aspheric diffractive multifocal and aspheric IOL group (0.21(0.03, 1.17), 0.34(0.06, 1.74)) (Z(1)=-0.10,-1.81, P(1)=0.92, 0.07).The mean spherical aberration for 3.0mm and 5.0mm optical zone in aspheric diffractive multifocal IOL group (0.01(-0.01, 0.20), 0.03(-0.10, 0.20)) were significantly lower than that in spherical IOL group (0.29(0.10, 0.99), 0.32(0.10, 0.99)) (Z(2)=-8.48,-8.54, P(2)=0.00, 0.01).But there was no statistical differences between the aspheric diffractive multifocal and aspheric IOL group (0.02(-0.09, 0.37), 0.04(-0.09, 0.37)) (Z(1)=-0.60,-0.73, P(1)=0.55, 0.46).About 86% of patients in aspheric diffractive multifocal IOL group do not need to wear glasses, it was better than the other two groups (χ(2)=17.83, 24.45, P=0.00, 0.00).The incidence of night glare and halo in aspheric diffractive multifocal IOL group 16/50(32%) was higher than that of aspherical IOL group 5/50(10%) and spherical IOL group 3/50(6%), and the difference was statistically significant (χ(2)=7.29, 10.98, P=0.00, 0.00).The overall satisfaction in aspheric diffractive multifocal IOL group was 45/50 (90%), better than that of aspherical IOL group 29/50(58%) and spherical IOL group 20/50(40%), and the difference was statistically significant (χ(2)=13.31, 27.47, P=0.00, 0.00). Conclusions: The aspheric diffractive multifocal IOL can provide patients with good and stable far, medium and near vision, to meet the needs of patients without glasses.At the same time, it can effectively reduce the high order aberrations and spherical aberration, improve visual quality.But due to night glare and glow, it does not apply to professional drivers and nighttime drivers. (Chin J Ophthalmol, 2017, 53: 599-609).

[Salubrinal protects human lens epithelial cells against endoplasmic reticulum stress-associated apoptosis].

OBJECTIVE: To study the protective effect of Salubrinal on human lens epithelial cells and its mechanism in endoplasmic reticulum stress (ERS). METHODS: Hydrogen peroxide (H2O2 200 µmol/L) was used to intervene in the cultured human lens epithelial cells B3 (HLE-B3) so as to create an oxidative stress model and induce ERS in the model. Different concentration of Salubrinal (10, 15, 20, 25, 30 and 35 µmol/L) were added to the cultured HLE-B3 with or without H2O2 intervention. Then the cells were cultured for 24 hours. The cell counting kit (CCK-8) assay was used to test the viability of HLE-B3. The HLE-B3 cells were divided into three groups: Group A (normal control group), Group B (H2O2 200 µmol/L group), and Group C (H2O2 200 µmol/L+ Salubrinal 25 µmol/L group). After 48 h, TUNEL and flow cytometry assay (FCM) were used to examine the effect of Salubrinal on HLE-B3 apoptosis. The expression of glucose-regulated protein 78(GRP78), C/EBP homologous protein (CHOP), cysteinyl aspartate specific proteinase 12 (Caspase-12) and phosphorylation eukaryotic translation initiation factor 2α (p-elF2α) were tested by western blot at different points in time. Data from different groups was analyzed by one-way analysis of variance (ANOVA) while Dunnett t test was used under an equal condition, Dunnett's T3 for the unequal variances. RESULTS: CCK-8 results showed that without the intervention of H2O2, different concentrations of Salubrinal had no inhibitive effect on HLE-B3 viability, and that survival rates were (98.6±3.3) %, (98.7±2.6) %, (99.4±3.2) %, (98.6±1.9) %, (98.8±2.5) %, (99.3±3.2) % and (99.5±2.4) %. There was no statistically significant difference between them (F=0.09, P=0.10). With the increasing of Salubrinal concentration, the survival rates of HLE-B3 in the presence of H2O2 intervention were (52.9±4.7) %, (65.0±3.6) %, (72.9±3.8) %, (84.5±3.6) %, (91.6±2.1) %, (93.1±2.9) %, (92.0±3.3) %. There was statistically significant difference from the control group (all P<0.01). However, the survival rates no longer increased (P=0.56, 0.88) if the Salubrinal concentration was greater than 25 µmol/L. FCM results indicated that apoptosis rates of Group A, B and C were (1.9±0.7) %, (8.8±0.5) %, (4.3±0.3) %, respectively and the differences were statistically significant (F=396.26, P<0.01, comparing with Group A, all P<0.01). TUNEL results showed that apoptosis indexes of Group A, B, and C were (7.7±1.0) %, (36.9±1.0) %, (16.7±2.2) %, respectively and the differences were statistically significant. (F=618.39, P<0.01, comparing with Group A, all P<0.01). RESULTS of western blotting in group B at different points in time (0, 12, 24, 36, 48 h) showed that p-elF2α had increased by (2.16±0.38) times at 6 h; GRP78 had increased by (2.56±0.15) times at 12 h; CHOP started to rise after 12 h until it dropped after 24 h, and its amount had increased by (2.49±0.23) times at 48 h; Caspase-12 had increased significantly by (3.53±0.30) times at 48 h. The expression of GRP78, CHOP and p-elF2α in group C was greater than that in Group B, but the expression of Caspase-12 in Group C was lower than that in Group B (GRP78: F=37.85, P<0.01; CHOP: F=61.09, P<0.01; Caspse-12: F=22.27, P<0.01; p-eIF2α: F=15.11, P<0.01). CONCLUSION: Salubrinal might protect HLE-B3 against H2O2-induced apoptosis by inhibiting ERS related apoptosis pathways.(Chin J Ophthalmol, 2016, 52: 437-443).

HOW EXPRESSIONS OF CLAUDIN-1 AND MMP-2 IN RETINOBLASTOMA CORRELATE WITH HISTOLOGICAL DIFFERENTIATION AND OPTIC NERVE INVASION.

Retinoblastoma is a commonly seen and dangerous intraocular malignant tumor in infants. Studies have found that Claudin-1 and MMP-2, whose expressions may be connected, play roles in tissues of retinoblastoma. In this study we analyze and discuss changes of Claudin-1 and MMP-2 expressions, and the correlation between the expressions and retinoblastoma histological differentiation and optic nerve invasion. MaxVisionTM was applied to detect expressions of Claudin-1 and MMP-2 in 45 samples of retinoblastoma and 15 paraffin-embedded samples of normal retina. The correlation between Claudin-1 expression and MMP-2 expression was analyzed based on chi-squared test and Spearman’s correlation test. Positive expressions of Claudin-1 in retinoblastoma were fewer than those in retina; higher positive expressions were found in differentiated tissues than in undifferentiated tissues; while compared to expressions in invasive optic nerves, Claudin-1 expressed more positively in optic nerves without invasion. As for MMP-2, its expressions were higher in retinoblastoma than in normal retina; undifferentiated tissues had higher positive expressions than differentiated tissues, which were not statistically significant; higher positive expressions were detected in invasive optic nerves. Thus, it could be concluded that the correlation between Claudin-1 expression and MMP-2 expression in retinoblastoma was negative. Expressions of Claudin-1 were positively related to histological differentiation and optic nerve invasion of retinoblastoma; while MMp-2 expression had negative correlation with histological differentiation and optic nerve invasion of retinoblastoma. Claudin-1 and MMP-2 played a negative role in the optic nerve invasion and tumor development of retinoblastoma.

Role of glutathione and other antioxidants in the inhibition of apoptosis and mesenchymal transition in rabbit lens epithelial cells.

This study aimed to discuss the effect and mechanism of glutathione (GSH) and catalase (CAT) on transforming growth factor beta (TGF-ß)-induced lens epithelial cell (LEC) apoptosis and epithelial mesenchymal transition (EMT) to prevent and control cataracts. Healthy rabbits (4 weeks old) were randomly selected, and LECs from their lenses were cultured in vitro. The 2nd- and 3rd-generation cells were divided into 6 groups (group A: 75 pg/mL TGF-ß2; B: 75 pg/mL TGF-ß2+10 mM GSH; C: 75 pg/mL TGF-ß2+300 U/mL CAT; D: 10 mM GSH; E: 300 U/mL CAT; and F: control group). Cell morphology was observed under an inverted microscope. The gap between cells increased, and the cells became reticulate after adding 75 pg/mL TGF-ß2; also, the cells swelled and appeared spindle-shaped. However, antioxidants reduced these changes. Growth inhibition was analyzed at 12, 24, and 48 h, and the differences between groups were not statistically significant. Cell apoptosis was analyzed, and the differences between group A and groups B and C were statistically significant (P<0.05). Reverse transcriptase-polymerase chain reaction was used to detect mRNA expression of α-SMA. The α-SMA mRNA level was greater in group A than in groups B and C (P<0.05). TGF-ß2 inhibited LEC proliferation and induced apoptosis and EMT. GSH and CAT inhibited apoptosis and EMT in LECs, and they had little effect on cell proliferation. Reactive oxygen species may be involved in cell apoptosis and EMT induced by TGF-ß2 as a cell-signaling molecule.

Mutation p.S335X in GATA4 reduces its DNA binding affinity and enhances cell apoptosis associated with ventricular septal defect.

Genetic mutations in GATA4, a transcriptional factor, have been found to cause congenital heart diseases. The underlying mechanism, however, remains largely unknown. We previously reported 7 heterozygous variants in patients with ventricular septal defects (VSD). Here we functionally characterized a de novo mutation p.S335X and demonstrated that this mutation led to the pre-termination of its translation, producing a truncated GATA4 lacking a conservative region at C-terminus. Truncated GATA4 did not disturb its subcellular localization; however, it delayed the cardiomyocyte differentiation in P19cl6 model and prohibited Bcl2 expression that led to apoptosis proved by fragmented genomic DNA and positive TUNEL staining in H9C2 cells. By ChIP assay, we showed that GATA4 without C-terminus reduced its DNA binding affinity and suppressed the expressions of its target genes. These findings suggest that C-terminus of GATA4 is critical to maintain DNA binding, and genetic mutations in this region may affect genes important for myocyte apoptosis and differentiation associated with congenital heart defects.

Identification of a locus for porokeratosis palmaris et plantaris disseminata to a 6.9-cM region at chromosome 12q24.1-24.2.

BACKGROUND: Porokeratosis palmaris et plantaris disseminata (PPPD) is a rare autosomal dominant dyskeratotic disorder characterized by a cornoid lamella with parakeratosis, hyperkeratosis and loss of granular layers. The genetic basis of this disease is still unknown. Two loci for disseminated superficial actinic porokeratosis (DSAP) were found to be located on 12q23.2-24.1 and 15q25.1-26.1. Both PPPD and DSAP are disseminated types of porokeratosis. OBJECTIVES: To locate the locus for PPPD, thereby facilitating the identification of this disease gene and leading to an understanding of the pathogenesis of porokeratosis. METHODS: Genotyping was performed in a Chinese family with PPPD using polymorphic microsatellite markers on 12q and 15q. RESULTS: The locus for PPPD is located within a 6.9-cM region between markers D12S1613 and D12S1341, with a maximum two-point LOD score of 8.14 (theta = 0.00) at D12S1335. CONCLUSIONS: This study provides a map location for isolation of a gene causing PPPD.

Phenotypic recombination between bean common mosaic and bean common mosaic necrosis potyviruses in vivo.

The US-5 strain of bean common mosaic virus (BCMV) and the NL-8 strain of bean common mosaic necrosis virus (BCMNV) are both seedborne potyviruses in common bean (Phaseolus vulgaris L). They have contrasting and highly stable biological characteristics which are genetically controlled. BCMV strain US-5 belongs to pathogenicity group IV. BCMNV strain NL-8 belongs to pathogenicity group III. The two strains have contrasting serological characteristics: NL-8 is serotype A; US-5 is serotype B. When these two strains were maintained separately or as a mixture for more than three years (39 serial transfers) or in more than 100 plants of either of two susceptible hosts, we were unable to isolate a single virus strain that exhibited mutant-like or recombinant-like characteristics. However, within 28 days (during the 1st passage) after these 2 strains were inoculated to opposite primary leaves of bean plants that were susceptible to one virus and resistant to the other, we were able to recover 17 strains that clearly possessed recombinations of various phenotypic characteristics from each of the two "parental" viruses. Three types of phenotypic characteristics were recombined singly or in combination during a single passage in vivo: 1) Biological characteristics known to be controlled by genes for pathogenicity; 2) Serotype; and 3) Temperature-induced hypersensitive vascular necrosis. Each of the phenotypic recombinant strains contained only pathogenicity genes or serological characteristics found in one or both parents. In no case did we isolate a strain that could be described as a random mutation or one that contained pathogenicity or serological characteristics which were not found in at least one parent strain. This is the first known demonstration of phenotypic recombinations between distinct potyviruses in vivo. Implications for the evolution of new virus strains through the use of resistant cultivars and its impact on breeding programs and bean seed production are discussed.