Analysis of the relationship between serum lncRNA ANRIL and miR-423-5p and airway inflammation and remodeling in children with bronchial asthma and their predictive value / 国际检验医学杂志

Objective To analyze the relationship between serum long chain non coding ribonucleic acid(ln-cRNA)ANRIL,microRNA(miR)-423-5p and airway inflammation and remodeling in children with bronchial asthma and its predictive value.Methods A total of 98 children with bronchial asthma treated in Haikou Ma-ternal and Child Health Hospital from June 2020 to December 2022 were selected.46 children with acute at-tack were selected as the attack group and 52 children with clinical remission were selected as the remission group.Another 50 children who were healthy during physical examination in the same period were selected as the health group.The relative expression levels of serum lncRNA ANRIL and miR-423-5p were detected by real-time fluorescence quantitative polymerase chain reaction.The serum inflammatory factor indicators[in-terleukin-13(IL-13),transforming growth factor-β1(TGF-β1),vascular endothelial growth factor(VEGF)]were detected by enzyme-linked immunosorbent assay.Airway remodeling indicators[bronchial thickness(T/D),pipe wall area/total cross-sectional area of gas pipeline(WA)]and lung function indicators[first second forced expiratory volume(FEV1),peak expiratory flow(PEF),maximum mid expiratory flow(MMEF)]were measured.The correlation between expression of serum lncRNA ANRIL,miR-423-5p and airway inflam-mation and remodeling indicators were analyzed by the Pearson method.The predictive value of serum ln-cRNA ANRIL and miR-423-5p in the diagnosis of bronchial asthma was analyzed by receiver operating charac-teristic(ROC)curve.Results The relative expression level of serum lncRNA ANRIL in remission and attack groups was higher than that in healthy group,and the relative expression level of serum miR-423-5p was lower than that in healthy group,with statistical significance(P＜0.05).The relative expression level of serum ln-cRNA ANRIL in the attack group was higher than that in the remission group,and the relative expression lev-el of serum miR-423-5p was lower than that in the remission group,with statistical significance(P＜0.05).The levels of serum VEGF,IL-13 and TGF-β1 in attack and remission groups were higher than those in healthy group,and the difference was statistically significant(P＜0.05).The levels of serum VEGF,IL-13 and TGF-β1 in attack group were higher than those in remission group,and the difference was statistically sig-nificant(P＜0.05).The levels of T/D and WA in the remission and attack groups were higher than those in the healthy group,and the levels of FEV,,PEF and MMEF were lower than those in the healthy group,with statistical significance(P＜0.05).The levels of T/D and WA in the attack group were higher than those in the remission group,and the levels of FEV1,PEF and MMEF were lower than those in the remission group,with statistical significance(P＜0.05).The results of Pearson correlation analysis showed that serum ln-cRNA ANRIL expression was positively correlated with airway inflammation and remodeling indicators,and negatively correlated with lung function indicators(P＜0.05).The expression of miR-423-5p was negatively correlated with airway inflammation and remodeling indexes,and positively correlated with lung function inde-xes(P＜0.05).ROC curve analysis showed that the area under the curve of lncRNA ANRIL and miR-423-5p alone and combined detection were 0.772,0.707 and 0.865 respectively,the predictive value of combined de-tection in diagnosing bronchial asthma was higher.Conclusion The relative expression level of serum lncRNA ANRIL increase in children with bronchial asthma,and miR-423-5p decrease,which promote airway inflamma-tion,remodeling,lung function decrease,and which has high diagnostic efficacy for children with bronchial asthma.

Investigation of the tissue-damaging effects and mechanisms of neutrophil-derived MMP-8 in Fusarium keratitis / 中华实验眼科杂志

Objective:To investigate the mechanism of tissue damage caused by neutrophil matrix metalloproteinase-8 (MMP-8) in Fusarium keratitis. Methods:A total of 108 male C57BL/6J SPF grade mice, 6-8 weeks old, were selected to establish a model of Fusarium keratitis (FK) in the right eyes.Corneal inflammation in mice was observed and scored under a slit lamp microscope.Based on the corneal inflammation scores, the modeling eyes were divided into 0, 12, 24, 48, and 72-hour groups post-modeling.At the corresponding time points, mice were euthanized, and corneal tissues were collected.The expressions of MMP-8, adenylate-activated protein kinase (AMPKα) and its serine 172-site phosphorylated form (p-AMPKα) proteins in corneal tissues were detected by Western blot.The neutrophil count in mice corneal tissues at each time point was determined using hematoxylin and eosin staining.The co-localization of neutrophils and MMP-8 protein in the cornea was observed by immunofluorescence staining.In the in vitro corneal collagen degradation experiment, corneal tissues were divided into MMP-8 group, buffer group, and normal saline group, which were treated with 100 μl of activated recombinant MMP-8, detection buffer, and normal saline, respectively.Hydroxyproline content in corneal tissues was determined using a hydroxyproline assay kit, and the mass fractions of hydroxyproline were compared among the groups.Peripheral blood neutrophils were isolated from human blood samples, and Fusarium spores were collected for experiments.Human neutrophils were divided into four groups, negative control group (cultured neutrophils), co-culture group (neutrophils co-cultured with spores), AICAR-treated group (neutrophils co-cultured with spores and treated with p-AMPK protein kinase activator AICAR), and compound C-treated group (neutrophils co-cultured with spores and treated with the inhibitor compound C).The MMP-8 protein expression levels in each group of human neutrophils were assessed via immunofluorescence staining.The use and care of animals complied with the ARVO statement and Regulations for the Administration of Affairs Concerning Experimental Animals.The animal experiment protocol was approved by the Animal Ethics Committee of Henan Eye Hospital (No.HNEECA-2017-04-02).One healthy adult volunteer was selected and 10 ml of peripheral venous blood was collected.The clinical study protocol was approved by the Clinical Ethics Committee of Henan Eye Hospital (No.HNEECKY-2019[16]). Results:At 24 hours post-modeling, corneal opacification was observed in the modeling eyes, and corneal perforation occurred in 72-hour post-modeling group.The corneal inflammation scores in 24, 48, and 72-hour post-modeling groups were all higher than those in 12-hour post-modeling group, and the differences were statistically significant (all at P<0.001).The relative expression levels of MMP-8 protein in the cornea were higher in 12, 24, and 48-hour post-modeling groups compared to 0-hour group, with statistically significant differences (all at P<0.001).There was a moderate positive correlation between the relative expression level of MMP-8 protein in the cornea and the inflammation scores of the modeling eye ( rs=0.50, P<0.05).In the cornea, the p-AMPKα (Thr 172)/AMPKα ratio was higher in 24, 48, and 72-hour post-modeling groups than in 0-hour group, and the differences were statistically significant (all at P<0.05).The p-AMPKα(Thr 172)/AMPKα ratio in the cornea was moderately positively correlated with the relative expression level of MMP-8 protein ( r=0.54, P<0.01).The number of neutrophils in the cornea was significantly higher in 24, 48, and 72-hour post-modeling groups than in 0-hour group, with statistically significant differences (all at P<0.001).The number of neutrophils in the cornea was strongly positively correlated with the inflammation score ( rs=0.77, P<0.001), and was moderately positively correlated with the relative expression level of MMP-8 protein ( r=0.56, P<0.05).MMP-8 protein expression in the cornea of the modeling eyes showed a high degree of co-localization with neutrophils.The hydroxyproline content in the cornea was (0.52±0.02)μg/mg, (0.51±0.03)μg/mg, and (0.27±0.02)μg/mg in buffer group, normal saline group and MMP-8 group, respectively, with a significant overall difference among them ( F=156.63, P<0.01).The corneal hydroxyproline content was lower in MMP-8 group compared to buffer and normal saline groups, and the differences were statistically significant (all at P<0.05).In the experiment involving the infection of cultured Fusarium spores with human neutrophils, the fluorescence intensity of MMP-8 expression was significantly higher in AICAR-treated group than in negative control group and compound C-treated group, with statistically significant differences (all at P<0.05). Conclusions:The MMP-8 secreted by neutrophils in mice with fungal keratitis can degrade corneal stromal collagen fibers, leading to corneal opacification or perforation.The variations in MMP-8 protein expression levels in human neutrophils may be associated with AMPK activation.

Monitoring results of national brucellosis monitoring sites in Jinzhong City, Shanxi Province from 2013 to 2021 / 中华地方病学杂志

Objective:To analyze the monitoring results of national brucellosis monitoring sites in Jinzhong City, Shanxi Province, and to provide a basis for scientific evaluation of prevention and control effects.Methods:From the "Disease Surveillance Information Report Management System", the surveillance results of national brucellosis surveillance sites (Pingyao County) in Jinzhong City from 2013 to 2021 were summarized and descriptive analysis was carried out.Results:From 2013 to 2021, a total of 996 cases of brucellosis were reported at the national brucellosis monitoring sites in Jinzhong City. A total of 3 538 serum samples were collected from key occupational groups, of which 161 were positive for brucellosis, with a positive rate of 4.55%. Among them, new cases accounted for 55.28% (89/161) and latent infections accounted for 21.74% (35/161). The annual serological test positive rate fluctuated between 1.19% and 20.55%. Except for 2014, the positive rate in all other years was less than 5.00%, and there was a statistically significant difference in positive rates between different years (χ 2 = 222.89, P < 0.001). The monitoring sites covered all townships within the jurisdiction (5 towns and 8 townships), and there was a statistically significant difference in positive rates among different townships (χ 2 = 26.45, P = 0.009). There was a statistically significant difference in positive rates among different occupations (χ 2 = 27.35, P < 0.001), with higher positive rates for veterinarians (5.41%, 2/37) and herders (4.76%, 29/609), respectively. There was a statistically significant difference in positive rates among different age groups (χ 2 = 42.97, P < 0.001), with a higher positive rate of 7.04% (104/1 477) in the 50 to 59 year old group. It can be seen from the scatter plot that the reported incidence rate did not increase linearly with the increase of serological positive rate. Conclusions:Except for 2014, the positive rate of serological tests for brucellosis at the national brucellosis monitoring sites in Jinzhong City, Shanxi Province has remained basically stable in other years. All townships in Pingyao County have an epidemic of brucellosis, and the 50 - 59 year old group and high-risk occupational groups are susceptible to it. The representatives of the monitoring results is not good, and it is recommended that the monitoring sites should strictly control the inclusion of the monitoring objects.

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.

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.

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.

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 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.

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.

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.

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.

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.

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.

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.

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.