Effects of Fusarium solani on AMPK phosphorylation and IL-6 expression in corneal epithelial cells / 中华实验眼科杂志

Objective:To investigate the expression of adenosine 5'-monophosphate-activated protein kinase (AMPK) phosphorylation in corneal epithelial cells and the effects of fungus on AMPK phosphorylation and interleukin-6 (IL-6) production in corneal epithelial cells.Methods:The human immortalized corneal epithelial cell line was selected.The safe concentration range of AMPK agonist 5-aminoimidazole-4-carboxamide 1-β-D-ribofuranoside (AICAR) (100, 300, 500, 1 000 μmol/L) and inhibitor Compound C (10.0, 12.5, 15.0, 17.5, 20.0 μmol/L) on corneal epithelial cells was screened by multi-function real-time unlabeled cell analyzer.Corneal epithelial cells without any treatment were used as the normal control group, and those co-cultured with spores were used as the spore control group.Corneal epithelial cells co-cultured with spores were treated with AICAR and Compound C for 4 hours in the AICAR group and Compound C group, respectively.The expression of phosphorylated AMPK (p-AMPK) and AMPK in corneal epithelial cells was detected by Western blot, and the concentration of IL-6 in the culture supernatant was determined by enzyme-linked immunosorbent assay (ELISA).Results:After treatment with different concentrations of AICAR for different periods, there was no statistical significance in the cell index of corneal epithelial cells (all at P>0.05). The cell index of corneal epithelial cells was increased with 10.0 μmol/L and 12.5 μmol/L Compound C treatment compared with that of the normal control group.The expression levels of p-AMPK were 0.67±0.15, 2.57±0.12, 3.67±0.58 and 1.50±0.50, respectively, in the normal control group, spore control group, AICAR group and Compound C group, showing a statistically significant difference among them ( F=32.820, P<0.001). The expression level of p-AMPK was significantly higher in the spore control group compared with the normal control group ( P<0.001). The expression level of p-AMPK in the AICAR group was higher than that in the spore control group, and the expression level of p-AMPK in the Compound C group was lower than that in the spore control group, and the differences were statistically significant (both at P=0.010). There was no significant difference in the relative expression level of AMPK among the four groups ( F=0.120, P=0.950). The expression levels of IL-6 concentration in the normal control group, spore control group, AICAR group and Compound C group were (107.81±17.15), (156.32±9.94), (167.96±14.16) and (127.42±19.75)pg/ml, respectively, showing a statistically significant difference among them ( F=15.210, P<0.001). The IL-6 concentration of the spore control group was higher than that of the normal control group, and the difference was statistically significant ( P<0.001). The IL-6 concentration of the AICAR group was higher than that of the spore control group, but the difference was not statistically significant ( P=0.260). The IL-6 concentration of the Compound C group was lower than that of the spore control group, and the difference was statistically significant ( P=0.010). Conclusions:In corneal epithelial cells, AMPK phosphorylation is found, which is enhanced after fungal spores stimulation, and the secretion of IL-6 increases.

Determination of tin and its compounds in ambient PM2.5 by atomic fluorescence spectrometry after ultrasonic extraction with concentrated hydrochloric acid / 环境与职业医学

Background Tin and its compounds can cause serious harm to human respiratory system and nervous system, but there is no corresponding national standard method for the determination of tin in PM2.5. Objective To establish a method for the determination of tin and its compounds in PM2.5 by atomic fluorescence spectrometry (AFS) after ultrasonic extraction with concentrated hydrochloric acid. Methods We extracted a fixed volume of air at a constant speed through a sampler with preset cutting characteristics to trap PM2.5 in the ambient air on quartz filter membranes. By selecting extraction solvent, comparing extraction temperature and time, and adjusting the acidity of solution to be measured, the sample pretreatment process was optimized, and a method for the determination of tin and its compounds in PM2.5 by AFS was proposed, and its performance indexes such as linearity, detection limit, and lower limit of quantification were obtained. The accuracy and precision of the method were evaluated by the standard addition recovery test with blank quartz filter membranes, and the interference test was carried out by adding standard urban particulate samples. The proposed method and the method recommended by the “Handbook on Monitoring and Protection of Air Pollution (Haze) Effects on Population Health (2020)” (the Handbook) were applied to actual samples, and the results were compared. Results This experiment used concentrated hydrochloric acid as the extraction solvent. The higher the reaction temperature and the longer the reaction time, the higher the recovery rate. Therefore, 70 ℃ water bath ultrasonic extraction for 3 h was selected. In terms of the proposed method, the linear range of detection was from 5.00 μg·L−1 to 50.00 μg·L−1, with a correlation coefficient ≥0.999 and a detection limit of 0.27 μg·L−1. When the quantitative detection of the lower limit was 0.90 μg·L−1，and the sampling volume was 144 m3, the limit of quantification was 1.25 ng·m−3. The recovery rate of standard addition of blank quartz filter membranes was 94.1%-97.5%, with a relative standard deviation ≤3.2%; the recovery rate of standard addition of standard urban particulate matter samples was 93.5%-103.0%, and the relative standard deviation was ≤2.1%, indicating that coexisting components in PM2.5 samples would not affect the determination of tin. For the 10 quartz filter membrane samples of PM2.5 monitoring, the results of tin by the established method (extraction with concentrated hydrochloric acid) were higher than those of the Handbook recommended method (extraction with nitric acid), and the difference is (3.61±0.54) ng·m−3(t=21.303, P<0.05). Conclusion The established method for the determination of tin and its compounds in PM2.5 by AFS after ultrasonic extraction with concentrated hydrochloric acid is simple, accurate, and suitable for laboratory determination of tin and its compounds in large quantities of PM2.5 samples.

Genetic diagnosis of a fetus with Dandy-Walker syndrome / 中华医学遗传学杂志

OBJECTIVE@#To explore the genetic basis for a fetus with Dandy-Walker malformation.@*METHODS@#G-banding chromosomal karotyping, single nucleotide polymorphism microarray (SNP array) and fluorescence in situ hybridization (FISH) were carried out for the fetus. Chromosomal karyotyping and FISH assay were also carried out for both parents.@*RESULTS@#SNP array has detected a 4266 kb microdeletion at 6p25.3p25.1 in the fetus, which was confirmed by FISH. FISH analysis of the parents demonstrated that the father has carried a cryptic t(6;14) (p25.1;p13) translocation, while the fetus has a der(6)t(6;14)(p25.1;p13) derived the paternal translocation.@*CONCLUSION@#The der(6)t(6;14)(p25.1;p13) probably underlies the Dandy-Walker malformation in the fetus. The 6p25.3p25.1 microdeletion is due to unbalanced gametes produced by the father's cryptic balanced translocation.

Genetic analysis of a child with Sotos syndrome / 中华医学遗传学杂志

OBJECTIVE@#To explore the genetic basis for a child with mentally retardation.@*METHODS@#G-banding karyotyping, single nucleotide polymorphism array (SNP-array) and fluorescence in situ hybridization (FISH) were performed for the child. Karyotyping and FISH were also carried out for her parents.@*RESULTS@#SNP-array has detected a 5077 kb microdeletion at 5q35.2q35.3 and a 4964 kb microduplication at 7q36.2q36.3 in the child. The results were confirmed by FISH. Based on above results, the father was subsequently found to carry a cryptic t(5;7) (q35.2; q36.2) translocation. The child was verified to have inherited a der(5) t(5;7)(q35.2; q36.2) from her father.@*CONCLUSION@#The 5077 kb microdeletion at 5q35.2q35.3 may have predisposed to the Sotos syndrome in the child. SNP-array combined with G-banding karyotyping and FISH can help to detect cryptic chromosomal translocations among patients.

Genetic diagnosis of a fetus with Dandy-Walker syndrome / 中华医学遗传学杂志

Objective@#To explore the genetic basis for a fetus with Dandy-Walker malformation.@*Methods@#G-banding chromosomal karotyping, single nucleotide polymorphism microarray (SNP array) and fluorescence in situ hybridization (FISH) were carried out for the fetus. Chromosomal karyotyping and FISH assay were also carried out for both parents.@*Results@#SNP array has detected a 4266 kb microdeletion at 6p25.3p25.1 in the fetus, which was confirmed by FISH. FISH analysis of the parents demonstrated that the father has carried a cryptic t(6; 14)(p25.1; p13) translocation, while the fetus has a der(6)t(6; 14)(p25.1; p13) derived the paternal translocation.@*Conclusion@#The der(6)t(6; 14)(p25.1; p13) probably underlies the Dandy-Walker malformation in the fetus. The 6p25.3p25.1 microdeletion is due to unbalanced gametes produced by the father’s cryptic balanced translocation.

Study on risk factors of postpartum hemorrhage in elderly parturients after cesarean section / 中国基层医药

Objective To investigate the risk factors of postpartum hemorrhage in elderly parturients after cesarean section based on logistic regression analysis and to explore nursing strategies .Methods From January 2018 to March 2019, a total of 210 elderly women with cesarean section and postpartum hemorrhage in the Integrated Traditional Chinese and Western Medicine Hospital of Wenzhou were selected as the study group .And 420 elderly women who underwent cesarean section and did not have bleeding were selected as the control group ,which may cause postoperative bleeding.Factors were analyzed and multivariate logistic regression analysis was performed on the factors that caused significant effects.Results The incidence of pregnancy hypertension ,abortion history,uterine atony, macrosomia and placenta previa in the observation group was significantly higher than that in the control group (t＝4.118-83.345,P＝0.000-0.042).There were no statistically significant differences in gestational diabetes mellitus , postpartum hemorrhage,coagulation function and depression between the two groups (t＝0.006-0.788,P＝0.375-0.938).Multivariate logistic regression analysis showed that gestational hypertension , history of abortion, uterine atony,macrosomia and placenta previa were risk factors for postpartum hemorrhage in elderly parturients after cesarean section (OR＝2.114 -19.521,P ＝0.035 -0.001 ).Conclusion Pregnancy -induced hypertension,history of abortion,uterine atony,macrosomia and placenta previa are the main factors affecting postpartum hemorrhage in elderly women with cesarean section.This provides an effective basis for the nursing of cesarean section postpartum hemorrhage in elderly women.

Efficacy and safety of pazufloxacin mesylate eye drops for bacterial conjunctivitis:a multicenter randomized, double-masked, parallel controlled clinical trial / 中华实验眼科杂志

Objective To compare the clinical efficacy and safety following the topical application of pazufloxacin mesylate eye drops with levofloxacin eye drops (LOFX) for bacterial conjunctivitis.Methods A multicenter,randomized,investigator-masked,parallel-controlled clinical trial was performed.Five hundred and twenty eyes of 520 patients with bacterial conjunctivitis were enrolled from March to October 2008 in seven ophthalmic centers in China.The patients were randomized into trial group and control group in 3 ∶ 1 ratio with the 390 eyes in the trial group and 130 eyes in the control group.Pazufloxacin mesylate eye drops was topically used 4 times per day for consecutively 7-14 days in the trial group,and levofloxacin eye drops was used in the same way in the control group.Microbiological cultures for conjunctiva sac secretions and drug sensitive test were carried out before and at the end of the administration of eye drops.The signs and symptoms were observed and scored before treatment and 0,3,7 and 14days after treatment.The adverse events following the administration of the eye drops were evaluated and compared.Results The intention to treat analysis (ITT) showed that the cure rate was 59.38％ in the trial group and 60.47％ in the control group,with the effective rate 88.80％ and 86.05％,respectively,with an insignificant difference between the two groups (x2 =0.12,P =0.72).The clinically evaluable analysis (CE) exhibited that the cure rates were 63.48％ in the trial group and 63.87％ in the control group,with the effective rate 92.46％ and 88.24％,whichwas not significantly different between them (x2 =0.54,P=0.46).The modified-ITT analysis (mITT) showed that the cure rates were 60.57％ in the trial group and 62.07％ in the control group,with the effective rate 90.32％ and 88.51％,without significant difference between the two groups (P＞0.05).Based on microbiologically evaluable analysis (ME),the clinical cure rates were 63.71％ and 63.41％ in the trial group and control group,and the effective rates were 93.44％ and 90.24％,respectively.There was no significantly difference between the both groups (P＞0.05).In the trial group and control group,the bacterial eradication rate was 89.42％ and 90.80％ based on ITT,90.11％ and 92.77％ based on CE,respectively.There was no significant difference in incidences of adverse events following the administration of the drug between the trial group and control group,including ocular tolerance,burning sensation,pricking and itching (P =0.34).Conclusions The effectiveness and adverse response were resemble between Pazufloxacin mesylate eye drops and LOFV following the topical application for bacterial conjunctivitis,which indicate that Pazufloxacin mesylate eye drops is effective and safe.

Prenatal diagnosis of a fetus with Phelan-McDermid syndrome / 中华医学遗传学杂志

OBJECTIVE@#To diagnose a fetus with Phelan-McDermid syndrome (PMS) using various techniques.@*METHODS@#Single nucleotide polymorphism array (SNP Array), multiplex ligation-dependent probe amplification (MLPA), fluorescence in situ hybridization (FISH) were applied in conjunction for the prenatal diagnosis of the fetus.@*RESULTS@#SNP Array detected a 4.03 Mb microdeletion at 22q13.31q13.33 in the fetus, which was confirmed by FISH and MLPA. FISH analysis of the parents suggested that the 22q13.31q13.33 deletion has a de novo origin.@*CONCLUSION@#Combined use of various techniques can enable accurate prenatal diagnosis and genetic counseling.

Study on risk factors of postpartum hemorrhage in elderly parturients after cesarean section / 中国基层医药

Objective@#To investigate the risk factors of postpartum hemorrhage in elderly parturients after cesarean section based on logistic regression analysis and to explore nursing strategies.@*Methods@#From January 2018 to March 2019, a total of 210 elderly women with cesarean section and postpartum hemorrhage in the Integrated Traditional Chinese and Western Medicine Hospital of Wenzhou were selected as the study group.And 420 elderly women who underwent cesarean section and did not have bleeding were selected as the control group, which may cause postoperative bleeding.Factors were analyzed and multivariate logistic regression analysis was performed on the factors that caused significant effects.@*Results@#The incidence of pregnancy hypertension, abortion history, uterine atony, macrosomia and placenta previa in the observation group was significantly higher than that in the control group (t=4.118-83.345, P=0.000-0.042). There were no statistically significant differences in gestational diabetes mellitus, postpartum hemorrhage, coagulation function and depression between the two groups(t=0.006-0.788, P=0.375-0.938). Multivariate logistic regression analysis showed that gestational hypertension, history of abortion, uterine atony, macrosomia and placenta previa were risk factors for postpartum hemorrhage in elderly parturients after cesarean section (OR=2.114-19.521, P=0.035-0.001).@*Conclusion@#Pregnancy-induced hypertension, history of abortion, uterine atony, macrosomia and placenta previa are the main factors affecting postpartum hemorrhage in elderly women with cesarean section.This provides an effective basis for the nursing of cesarean section postpartum hemorrhage in elderly women.

Genetic analysis and prenatal diagnosis of a sporadic family with neurofibromatosis type 1 / 浙江大学学报·医学版

OBJECTIVE@#To identify pathogenic mutation for a family with neurofibromatosis type 1(NF1) and provide prenatal diagnosis for them.@*METHODS@#Mutation analysis of the sporadic family with NF1 was performed with target captured next generation sequencing and Sanger sequencing. RNA samples were extracted from the lymphocytes of NF1 patient and her parents. RT-PCR and Sanger sequencing were performed to analyze the relative mRNA expression in the samples. Prenatal diagnosis of the pathogenic mutation was offered to the fetus.@*RESULTS@#A novel splicing mutation c.1260+4A>T in the gene was found in the proband of the family, but was not found in her parents.cDNA sequencing showed that 13 bases inserted into the 3' end of exon 11 in the gene lead to a frameshift mutation. Prenatal diagnosis suggested that the fetus did not carried the mutant.@*CONCLUSIONS@#The : c.1260+4A>T mutation found in the NF1 patient is considered to be pathogenic, which provides information for family genetic counseling and prenatal diagnosis.

Genetic analysis of a fetus with multiple malformations caused by complex translocations of four chromosomes / 浙江大学学报·医学版

OBJECTIVE@#To conduct genetic analysis in a fetus with complex translocation of four chromosomes.@*METHODS@#G-banded chromosome karyotype analysis, single nucleotide polymorphism array (SNP array) and fluorescence hybridization (FISH) were performed in a fetus with multiple malformations. Peripheral blood chromosome karyotype and FISH were also carried out for the parents.@*RESULTS@#The fetal amniotic fluid karyotype was 46, XY, t(12; 13)(q22; q32). SNP array analysis showed that there were 20 192 kb duplication at 1q42.13q44 and 13 293 kb deletion at 15q26.1q26.3 in the fetus. The results of karyotype and SNP array were inconsistent. FISH analyses on the parental peripheral blood samples demonstrated that the mother was a cryptic 46, XX, t(1; 15)(q42.1; q26.1) translocation. The fetus had inherited 46, XY, t(12; 13)(q22; q32) from his father and der(15)t(1; 15)(q42.1; q26.1) from his mother.@*CONCLUSIONS@#The 1q42.13q44 duplication and 15q26.1q26.3 deletion may have contributed to the abnormal sonographic features of the fetus. The combination of cytogenetic, SNP array and FISH techniques was beneficial for providing an accurate genetic counseling.

Antifungal efficacy of corneal collagen cross-linking on treating experimentally induced fungal keratitis of mice / 中华实验眼科杂志

Objective This study was to assess the efficacy of corneal collagen cross-linking treatment on fungal keratitis.Methods Eighty SPF male C57BL/6 mice aged 6-8 weeks were selected for the experiment.Fusarium solani infected model was established on the left eyes of all 80 mice.Forty mice were distributed randomly into sham operation group,model control group,scraped epithelium group and corneal collagen cross-linking (CXL)group (treated with epithelium scraped and CXL).Three days after modeling,the levels of the corncal disease sevcrity were scored by slit lamp microscopy.The fungal activity was confirmed by plate counts.The left 40 mice were divided randomly into sham operation group,model control group,scraped epithelium group and CXL group (treated with epithelium scraped and CXL).In 1 day and 2,3,4,5,6,7,14 days after modeling,the corneas were examined under the slit lamp microscope.The corneal pathological examination of each group were conducted with hematoxylin and eosin staining at postoperative 14 days.The animal feeding and use was in accordance with the standards set by the ARVO,and the experiment was approved by the Ethic Committee for Experimental Animal of Henan Eye Institute.Results The colony-forming units (CFUs) of fungal solutions in culture significantly decreased with CXL treatment (F =11.97,P =0.00).The Pearson correlation analysis of CFU and clinical scores in CXL group showed that inflammatory cells infiltration was positively correlated with corneal disease severity (r =0.723,P =0.043).Corneal inflammatory score was significantly lower in the CXL group in various time points,with a significant differences among the groups and time points (Fgroup =34.44,P=0.00;Ftime =17.49,P=0.00).Corneal lesion and the depth of ulceration in scraped epithelium group and CXL group were remarkably lower than that in the model control group (all at P ＜ 0.05).Histopathology revealed that the degree of corneal collagen fibers destruction and the ratio of inflammatory cells infiltration in scraped epithelium group (59.33％) and CXL group (11.29％) were much lower than that in the model control group (73.65％).Conclusions CXL can inhibit the fungal activity effectively in the cornea of mice,and reduce the fungal induced keratitis reaction.

A retrospective cohort study on orthokeratology for high myopia progression / 中华实验眼科杂志

Objective To study the therapeutic effect and safety of orthokeratology in the teenagers with high myopia.Methods A retrospective cohort study was carried out.The clinical data of 120 eyes from 60 high myopia patients wearing orthokeratology contact lens for 1 year in Henan Eye Hospital from June 2013 to June 2014 was analyzed,and the data of contemporaneous 118 eyes from 59 matched high myopia patients wearing conventional glasses were used as controls.The initial spherical equivalent (SE) of all the patients was-6.00 to-9.00 D.The maximum correction amplitude of the orthokeratology contact lens was set as 6.00 D based on the Regulation of China Food & Drug Administration.The contact lenses were worn for 1 month firstly and then the conventional glasses were used at daytime and contact lenses were consecutively worn at night for 1 year in the contact lenses group,and only glasses were worn in the conventional glasses group.The uncorrective visual acuity (UCVA),SE,corneal fluorescein staining were examined 1 week,1 month,3 months,6 months and 12 months after wearing lenses.The changes of corneal endothelial cells were examined 6 and 12 months after wearing lenses.Results The UCVA was improved and SE was reduced 1 month and 3,6 and 12 months after wearing lenses in comparison with 1 week after wearing lenses in the contact lenses group (all at P＜0.01);while no significant change was seen in UCVA in the conventional glasses group (all at P＞0.05).Compared with wearing contact lenses,the SE was increased in various time points after wearing glasses in the conventional glasses group (all at P＜0.01).The axial length and SE extended by (0.08±0.12)mm and (0.19±0.21)D in the contact lenses group and those in the conventional glasses group were (0.29±0.14)mm and (0.69±0.27)D,showing significant differences between the two groups (t =10.024,5.691,both at P＜0.01).No significant differences were found in corneal endothelial cell density,percentage of hexagonal cells and coefficient of variation of corneal endothelial cells in various time points after wearing contact lenses (corneal endothelial cell density:Fgroup =0.090,P =0.769;Ftime =0.133,P =0.563.percentage of hexagonal cells:Fgroup =0.071,P=0.836;Ftime =1.091,P =0.203.coefficient of variation:Fgroup =0.107,P =0.734;Ftime =1.948,P =0.156).The incidence of corneal fluorescein staining was 31.67％ in the contact lens group,which was much higher than 3.51％ in the conventional glasses group (x2 =97.910,P =0.001).Conclusions Compared with wearing only conventional glasses,orthokeratology can improve UCVA and control axial length extension in juvenile with high myopia,but orthokeratology is more likely to damage the corneal superficial layer.

Feasibility of confocal laser scanning microscopy with slit-lamp microscope in the diagnosis of filamentous fungal corneal infection with culture negative patients / 中华实验眼科杂志

Objective To investigate the imaging manifestations of different filamentous fungal strains under the confocal laser scanning microscope and slit-lamp microscope,and evaluate the feasibility of rapid diagnosis and therapeutic efficacy judgment for fungal culture negative patients.Methods A diagnosis trail was performed.Nine hundred and ninety-three patients with fungal keratitis (FK) which were varified by fungal culture were included in Henan Eye Hospital from September 2013 to January 2014.Distribution of fungi strains and positive rate of fungal strains by fungal culture and corneal confocal laser scanning microscopy were compared.The imaging characteristics of different filamentous fungi and different stages of one filamentous fungi under the slit-lamp microscope and confocal laser scanning microscopy were summarized.Results In the 993 FK patients,the diagnostic positive rate of fungal culture and confocal laser scanning microscopy was 43.20％ and 82.07％,respectively,showing a significant difference between them (x2 =45.323,P =0.000).In 429 culture-positive patients,the diagnostic positive rate of confocal laser scanning microscopy was 92.31％;while in 564 culture-negative patients,the diagnostic positive rate of confocal laser scanning microscopy was 74.29％.In 429 culture-positive patients,Aspergillus was the most common genus,accounting for 50.12％,and followed by Fusarium sp.and Altemaria sp.(18.18％ and 10.49％).There were no significant differences in fungal species distributions between fungal culture and confocal laser scanning microscopy examination in 429 cases (all at P＞0.05).The imaging characteristics under the slit-lamp microscope and confocal laser scanning microscope were different in different fungi stains.Aspergillus infection showed a plume-like corneal ulcer,and the Aspergillus sp.hyphae were thin and line-shaped with high reflective light and less branched under the confocal laser scanning microscope.Toothpaste-like corneal infiltration was seen in Fusarium sp.-infectious lesions under the slit lamp microscope,and mycelium showed a high-reflective long rod-like image with less branch in the image of confocal laser scanning microscope.Alternaria alternate sp.corneal infection showed nevus lesions,and hyphae characterized by high-reflective long rod or string beads in shape with less branches in the image of confocal laser scanning microscope.The mycelium was ruptured,shorter,thinner with weak reflective light following drug therapy.The differential diagnosis could be easily obtained between hyphae and corneal nerve fibers by confocal laser scanning microscope.Hyphae intertwined,or had branches with diffuse distribution,which surrounded by highreflective inflammatory cells and destructed matrix fiber and were located in stroma.The corneal nerve fibers located between epithelium layer and stroma layer,surrounded by normal epithelium or stroma structure.The diameter of the thicker nerve fibers in the stroma layer was obviously thicker than that of the hyphae.Conclusions The diagnosis rate of confocal laser scanning microscope combined with slit-lamp microscope for filamentous fungi-infectious FK is higher than that of fungal culture.The combination procedure of confocal laser scanning microscope and slit lamp microscope examination provides a rapid evaluation for fungi strains and therapeutic efficacy in the FK patients with negative results by fungal culture.

Clinical analysis of 56 cases with vaginal intraepithelial neoplasia after hysterectomy / 上海交通大学学报（医学版）

Objective · To investigate the relationship between the results of thin prep cytologic test (TCT), high-risk human papillomavirus (Hr-HPV) detection and vaginalintraepithelial neoplasia (VAIN) after hysterectomy. Methods · A retrospective study was conducted of 56 patients with VAIN after hysterectomy. The analysis included TCT and Hr-HPV examination, clinical data and the relationship between Hr-HPV examinenation and histopathological examination of colposcopy. Results · Postoperative follow-up of TCT and Hr-HPV parallel pathological examination showed that 56 patients had vaginal stump lesions, including cervical factor hysterectomy accounted for 5.81% (45/775) and non cervical factor hysterectomy accounted for 0.19% (11/5933). The difference was statistically significant (P=0.000). In 56 cases of patients with vaginal stump lesions, Hr-HPV infection were 40 cases (accounting for 71.43%), uninfected patients were 16 cases (accounting for 28.57%). The Hr-HPV infection rates of cervical hysterectomy and non cervical factor hysterectomy patients were 80.00% (36/45) and 36.36% (4/11) respectively, and the difference was statistically significant (χ2= 6.248, P=0.012). TCT results showed that the incidence of squamous intraepithelial lesions were 42.22% (19/45) and 9.09% (1/11) respectively. Conclusion · The Hr-HPV infection rate and the morbidity of VAIN of the patients undergone hysterectomy due to the cervical lesionfactors is higher than those who had none cervical lesion factors. In order to identify VAIN early, patients who have the history of hysterectomy should undergo careful evaluation of cervical and vaginal circumstance before surgery and the routine examinations of TCT and Hr-HPV in the follow-up.

Research progress on the safety of offsprings conceived by assisted reproductive technology / 浙江大学学报·医学版

Assisted reproductive technology (ART) employs superovulation,culture and other micromanipulation to complete oocyte maturation, fertilization and early embryo development. Although these techniques have been successfully applied to solve infertility problems, the process may interfere in cell proliferation, differentiation and growth. The clinical and laboratory studies on the safety issue of ART are reviewed in this article. Studies found that the incidence of birth defects increased in ART offspring. Superovulation,culture and intracytoplasmic sperm injection may induce epigenetic aberrations during embryo development, which would influence the development of ART conceived children. The epigenetic susceptibility related to ART might be transmitted to subsequent generations, and the potential impact on ART offspring still need further investigation. In addition, ART treatments may also increase the risk of genetic diseases.

Effect of MMP-8 on cornea collagen / 重庆医学

Objective To investigate the effect of MMP-8 on cornea. Methods Fifteen C57BL/6J healthy mice were selected. The right eyes corneal stroma was injected by 10μL MMP-8 as the experimental group and the left eyes were injected by same amount of normal saline as the control group. At 0,4,8 h, the two-photon microscope second harmonic generation imaging technology was used to scan mice corneal stroma layer by layer in vivo. The obtained images were performed the 3D reconstruction by Imaris software and the signal intensity of the images were calculated. At 4,8 h, the corneal opacity degree was evaluated under slit lamp. At 8 h,mice were killed and corneas were collected to determine the hydroxyproline concentration. Results The cornea stromal fiber signal strengthes at 0 h in the experimental group and control group were (89.7±11.2) and (85.3±7.0),which at 4 h were (14.5±3.4) and (46.6±14. 0) respectively,which at 8 h were (11.0±4.6) and (34.6±12.5) respectively. The cornea stromal signal strength at 4,8 h in the experiemental group was significantly decreased compared with that in the control group (P<0.05) ;the cornea at 4 ,8 h in the experimental group was significantly turbid than that in the control group (P<0.05);the cornea hydroxyproline concentrations detected at 8h in the experiemental group and control group were (0.433±0. 090) μg/mg and (0. 590±0. 133) μg/mg respectively,the experimental group was significantly lower than the control group (F=7. 193,P=0. 014). Conclusion MMP-8 has obvious degradation and destroy effect on mice corneal stroma collagen,which leads to the decrease of corneal opacity.

Prognosis of clinical treatment for fungal keratitis and its influence factors / 中华实验眼科杂志

Background Fungal keratitis has poor prognosis and high blinding rate,so it is vital to identify the risk factors that affect the treating outcome and prognosis of fungal keratitis.Objective This study was to evaluate the clinical outcome of fungal keratitis and identify the affecting factors.Methods This was a series cases-observational study.A total of 267 eyes of 267 patients with fungal keratitis were included in Henan Eye Institute during January 2013 to January 2014.Eye examinations were performed including slit-lamp microscope examination,corneal smears,confocal microscopy,fungal culture and identification.Combined anti-fungal drugs were topically used in all eyes for the initial treatment then the treating regimen were selected based on the susceptibility testing.corneal lesion resection combined with conjunctival flap transplantation,lamellar corneal transplantation or penetrating corneal transplantation was performed respectively on the patients with poor curative effect after drug treatment.The clinical prognosis of the patients was graded and the affecting factors for clinical outcome were evaluated.This study protocol complied with Helsinki Declaration and was approved by Ethic Committee of Henan Eye Hospital.Written informed consent was obtained from patients or guardians.Results One hundred and eighty-five eyes were cured by anti-fungal eye drops with the effective rate of 69.29％,and the inflammation was controlled in 60 eyes who received combination procedure of anti-fungal drugs with different surgeries,while 22 eyes lost visual function because of refractory glaucoma and final evisceration.The total clinical effective rate was 91.76％.Aspergillus and Fusarium spp.appeared to be the most common pathogenic fungi by drug sensitivity test.The keratitis caused by Aspergillus had lowest cure rate among different fungi (x2 =11.350,P =0.002) and the most poor clinical prognosis (H=31.285,P =0.013).The medication curative rate was 71.8％ in the patients with positive culture outcome,which was higher than 62.5％ in the patients with negative culture outcome (x2=8.75,P＜0.01).A significant difference was found in the medication curative rate and prognosis between the patients with ≥ 2 kinds of sensitive anti-fungal drugs and the patients with ＜2 kinds of sensitive anti-fungal drugs (77.5％ versus 52.3％,x2 =9.63,P＜0.01;H=24.281,P =0.021).Size of infiltration area,with or without hypopyon,number of sensitive drugs,and pathogenic fungi were significantly correlated with clinical outcome (all at P ＜ 0.05).Conclusions In vitro susceptibility testing can guide clinical drug treatment of fungal keratitis.Infiltration area ＞ 16 mm2,hypopyon ＞2 mm,resistance to those drugs in vitro and infection of Fusarium.spp or Aspergillus.spp are predictors of a poor outcome.

Clinical effect of bioengineered cornea and in vivo confocal microscopy observation after human lamellar keraplasty / 中华实验眼科杂志

Background Xenotransplantation is arousing more attention of researchers because of the lack of corneal donors.Biological corneal scaffolds constructed by porcine corneal acellular stroma appears to have an acceptable biocompatibility.However,its clinical effects and the histomorphological features in the corneal tissue of receipts' are still unclear.Objective This study was to evaluate the viability of bioengineered corneas as a new material of human lamellar keratoplasty and observe the in viva biological features after human keratoplasty under the laser confocal microscope.Methods A prospective serial cases observational study was carried out.Fifteen eyes of 15 patients with infectious keratitis were enrolled in Henan Eye Hospital from February to August 2014 under the approval of Ethic Committee of Henan Eye Hospital and informed consent of each patient, including 4 eyes of fungal keratitis,1 eye of bacterial keratitis, 9 eyes of mixed infectious keratitis and 1 eye of leucoma.Corneal lamellar transplantation was performed on the eyes with the bioengineered corneas as grafts and the follow-up time was one year.The survival of grafts was assessed and scored, and the indices including corneal transparency,neovascularization, graft lysis or inflammatory recurrence and visual acuity (logMAR) were evaluated.The morphology and density of corneal epithelial cells and endothelial cells, corneal stroma and subepithelial neural fibers were examined by laser confocal microscope 3,6,9 and 12 months after surgery.Results Postoperative inflammatory response was seen 3 days and disappeared 7 days after surgery.The grafts were clear 1 month after surgery, and no corneal dissolution was found during the follow-up duration.Glaucoma occurred in 1 eye at 6 months and graft rejection occurred in another eye 12 months after surgery.The logMAR, corneal transparency scores and corneal neovascularization scores improved after surgery in comparison with before surgery,with a significant difference among various time points (x2 =92.63,59.37,10.50 ,all at P＜0.05).Complete epithelization of grafts was seen.Compared with the contralateral eyes,the morphology of epithelial cells was similar and endothelial cells were enlarged in the operated eyes.In addition,no stromal cell structure was seen in the corneal stroma in the operative eyes.Subepithelial nerve fibers appeared in 8 eyes at 6 months, but the fiber density was lower in the operated eyes than that in the contralateral eyes.Significant difference was found in epithelial cell density among different time points (F=1.48, P =0.22).The endothelial cell densities were (2 542 ± 119), (1 895 ± 129), (1 869 ± 135), (1 854 ± 101) and (1 844 ± 103)/mm2 before surgery and 3, 6, 9 and 12 months after surgery, showing significant differences between preoperation and postoperative time points (all at P＜0.05).The subepithelial nerve fibers densities were (1.26± 0.13),(3.62±0.81) and (5.98±0.44) mm/mm2 at 6,9 and 12 months after surgery,with significant differences between adjacent two time points (t'=-8.16 ,-7.24;both at P=0.00).Conclusions Bioengineered corneal grafts survive well in human eye after lamellar transplantation,which can reconstruct ocular surface and improve the visual acuity.Biological cornea can replace human corneal materials.

Safety and efficacy of 0.05％ tacrolimus eye drops for refractory ulcerative keratitis / 中华实验眼科杂志

Background Local medical treatment of refractory immunologic keratitis is unsuccessful,and systemic steroids and immunosuppressive agents could cause severe side-effects.Tacrolimus is a potent immunosuppressive drug,it has been proved that topical application of tacrolimus could reduce immunologic inflammation.The safety and efficacy of 0.05％ tacrolimus eye drops for refractory immunologic keratitis has not been described.Objective This study was to evaluate efficacy and safety of 0.05％ tacrolimus eye drops for refractory ulcerative keratitis.Methods A retrospective study was performed.Twenty-one eyes of 17 patients with refractory immunologic keratitis,which had uncontrolled inflammation despite initially treatment including topical steroids and 1％ cyclosporine A,were enrolled,including 11 males and 6 females,with the mean ages of 52 years.Infectious ulcer was excluded by laboratory tests.No systemic disease was found in 11 patients,and Wegener's granulomatosis,rheumatoid arthritis and ulcerative colitis were seen in 1 patient,4 patients and 1 patient respectively before presentation and they were all in remission under conventional systemic therapy.Four patients got binocularly involved and thirteen patients were monocularly involved.Of the 21 eyes,2 eyes with ulcer were ≥ 3 quarters of the limbus,and 19 eyes with ulcer were ≤ 2 quarters.All patients were treated with 0.05％ tacrolimus eyedrops after discontinuing cyclosporine A.The dosage was adjusted according to the severity of inflammation and was gradually tapered when improvement occurred.The corneal lesions were examined under the slit lamp microscope and Heidelberg HRT3 Rostock Cornea Module regularly,and inflammatory cell infiltrations were analyzed with Cell Count(R) software (Heidelberg Engineering GmbH).The safety variables were monitored regularly,including adverse response of eye,tacrolimus blood concentrations measured by chemiluminescent microparticle immunoassay (CMIA) and laboratory examinations of blood routine,blood glucose level,liver and kidney function.Results The patients were treated and followed-up for a mean duration of 18.1 months (range,8-24 months).Corneal ulcer area was obviously reduced 1 month after treatment in 19 eyes,and 2 eyes of 2 cases received anterior lamellar keratoplasty due to progressive corneal destruction despite of tacrolimus therapy.Corneal ulcer was cured 3 months after treatment,and stromal edema and infiltration disappeared 6 months after treatment under the slit lamp microscope.The inflammatory cell densities at lesion zone were (958±329),(858±339),(459±261),(192±124),(98±52),(44±24) and (3±2)/mm2 before treatment and 1 week,1 month as well as 3,6,12,24 months following treatment,respectively,showing a gradually decline as time lapse (F =125.439,P =0.000),and the inflammatory cells were significantly decreased in 1,3,6,12 and 24 months following the administration of 0.05％ tacrolimus eye drops in comparison with that before treatment (all at P =0.000).The therapy duration was 12 months in 9 eyes and 24 months in 12 eyes.Transient irritation sensation occurred in 4 eyes during the treating period.Blood concentrations of tacrolimus were below 1.0 ng/ml in all of the patients.No abnormality was found in laboratory tests.Conclusions The use of 0.05％ tacrolimus eye drops is a safe and effective approach to refractory immunologic keratitis.