Ophthalmic microsurgery has its own characteristics: high precision , complicated operation , and long learning curve.It is important for young ophthalmologists to master the basic operation skills through standardized and systematic training during graduate studies .The teaching method , integrating theoretical teaching , animal operation and simulative ophthalmic surgery training , have important meanings in promoting the post-graduate students to master the practical and correct ophthalmic microsurgery technique . The aim of this study was to introduce the curriculum design and practice , and to evaluate the model through the examination scores .

Objective To investigate the effects of hypoxia on the expression of Stat3 and p-Stat3 ,and assessed the apoptosis ability of JAR cells in vitro .Methods JAR cells were cultured under hypoxic conditions .Western blot were used to determine the protein expression of Stat3 and p-Stat3 .Cellular apoptosis was monitored by flow cytometry analysis .Results Abnormal morpholo-gy changes in trophoblast cells under low oxygen conditions .After 48 h hypoxic treatment ,the protein of Stat3 and p-Stat3 were significantly decreased(P< 0 .05) ;however ,the level of apoptosis was significantly increased (P < 0 .05) .Conclusion Stat3 and p-Stat3 protein levels were decreased under hypoxia circumstance ,while the cell apoptosis ability was increased in JAR cells .

Objective To investigate the hepatitis B virus (HBV) infection status quo among childbearing age women to provide the theoretical basis for adopting the effective intervention ,treatment and blocking measures in pre-pregnant women .Methods The enzyme-linked immunosorbent assay (ELISA) and the time-resolved fluorometric immunoassay were adopted to detect HbsAg in 68 682 children bearing age women ,including outpatient and hospitalized patients ,in this hospital from February 2012 to September 2013 .The detect results were analyzed according to the age ,place of residence and medical departments .Results Among 68 682 children bearing age women ,3 260 cases were found positive for HbsAg with the HBsAg positive rate of 4 .70% .The statistically significant differences in the HBV infection positive rate existed among different age groups ,the positive rates in the age groups of 16- <20 years ,20 - < 25 years ,25 - < 30 years ,30 - < 35 years ,35 - < 40 years ,40 - < 45 years and 45 - < 50 years were 3 .83% ,4 .89% ,4 .96% ,4 .91% ,4 .18% ,4 .40% and 4 .18% respectively ,the differences among them had statistical significance (χ2 =15 .76 ,P=0 .015) ,in which the age 25- <30 years group had the highest HBV infection rate .At the same time the signifi-cant differences in the geographical distribution and medical departments existed .The HBV infection positive rate in rural area was significantly higher than that in city with statistical difference (χ2 =27 .47 ,P<0 .05) ,and the HBV infection positive rate in the outpatient departments was higher than that in the inpatient departments with statistical difference (χ2 =46 .88 ,P<0 .05) .Conclu-sion Children bearing women at 20-34 years old show the highest HBV infection rate .The HBV screening and the works of pre-vention and treatment during this age period should be strengthened ,which conduces to discover the HBV infection positive patients in time and prevent the mother-to-fetus transmission .

The paper introduces the common methods for automatic de-identification of clinical texts,including the method based on rules,machine learning method,comprehensive method,etc.,states the challenges for clinical texts practicability,system universality and scalability of clinical texts de-identification research,analyzes the further research direction,and provides reference for researchers of this field.

Objective To observe the effect of small-interfering RNA(siRNA) targeting c-Met,the receptor of hepatocyte growth factor(HGF),on the proliferation of human lens epithelial cells(LECs).Methods siRNA was transferred into LECs cultured in vitro by HiperFect Transfection Reagent.Real-Time PCR was applied to observe the expression of c-Met mRNA in LECs after gene transfer,and MTT assay was used to detect the proliferation of LECs induced by HGF.Results The expression of c-Met mRNA in LECs was significantly decreased in the experimental group,compared to that in the controls(P<0.01).Proliferation of LECs induced by HGF was inhibited,compared with the single HGF stimulated group(P<0.01).Conclusion The RNA interference targeting c-Met can effectively inhibit the expression of c-Met mRNA,and the proliferation of LECs induced by HGF.

Objective To evaluate the feasibility of apply Ion Proton semiconductor sequencing platform in non-invasive prenatal genetic diagnosis .Methods Totally 1 000 pregnant women with a singleton pregnancy of 12-32 weeks gestation were selected from the Third affiliated Hospital of Zhengzhou University from Jan to Dec 2013.Using noninvasive prenatal genetic diagnosis based on Ion Proton semiconductor sequencing platform to study their cffDNA .In parallel, 72 pregnant women received invasive prenatal diagnosis by traditional chromosomal analysis with amniocentesis chorionic villus sampling .Results It′s shown that 18 out of 1 000 (1.8%) pregnant women underwent the noninvasive prenatal genetic testing had a high risk for aneuploid chromosomes , including 7 cases of 21-trisomy, 4 cases of 18-trisomy, 2 cases of 13-trisomy, 4 cases of sex chromosomal abnormality , and 1 case of 15-trisomy.It demonstrated that the rate and accuracy of fetal 21-trisomy, 13-trisomy and 18-trisomy by non-invasive prenatal genetic testing were both 100%without misdiagnosis , the rate of detection for sex chromosomal abnormality was 2/2 with a false positive rate of 1/3.However, the 15-trisomy predicted by the non-invasive prenatal diagnosis in a woman was finally proved to be a false positive .Based on the results by karyotyping (55/55) as well as follow-ups (493/493), the specificity of the non-invasive prenatal diagnosis for detection of 21-trisomy, 18-trisomy and 13-trisomy was 100%.One Ion PITM chip could detect 12 to 15 samples in 1.5 h and the whole process of noninvasive detection could be completed in 1 to 1.5 days.Conclusions The non-invasive prenatal diagnosis by Ion Proton semiconductor sequencing platform could provide fast and accurate detection of fetal aneuploidy .The benchtop high-throughput sequencing platform has laid the foundation for the independent application in clinical settings for fetal aneuploidy detection .

Objective:To investigate the changes in adaptive phenotypes of Yersinia pestis ( Yp) during successive passages in macrophages. Methods:A Yp strain of 201-MI was induced by 50 successive passages of Yp 201 strain in Raw264.7 cells. Phenotypic characteristics of 201 and 201-MI strains were compared by analyzing their survival rates in macrophages, growth curves, biofilm formation abilities, acid and hydrogen peroxide-stress tolerance, and virulence to mammal cells (Raw264.7 and HeLa cells) and mice. Results:Comparing with 201 strain, 201-MI strain showed various phenotypic changes, including higher survival rate in Raw264.7 cells, faster growth in iron-deficient medium, higher tolerance to acid and hydrogen peroxide, decreased biofilm formation ability, and less damages to Raw264.7 and HeLa cells. More-over, 201-MI strain showed decreased virulence to mice in both subcutaneous and intraperitoneal challenges. Preliminary comparative genomics analysis revealed some indel and nonsense mutations in 201-MI strain, which might account for its phenotype changes.Conclusions:After successive passages in macrophages, Yp showed some phenotypic changes, which might reflect its adaptive evolution under the pressure of macrophages. Detailed multi-omics analysis would be of great help to understand the underlying genetic mechanisms of these changes, and the related Yp-macrophage interaction processes as well.

The antigen gene expression level of a DNA vaccine is the key factor influencing the efficacy of the DNA vaccine. Accordingly, one of the ways to improve the antigen gene expression level of a DNA vaccine is to utilize a plasmid vector that is replicable in eukaryotic cells. A replicative DNA vaccine vector pCMVori was constructed based on the non-replicative pcDNA3.1 and the replicon of porcine circovirus 2 (PCV2) in this study. An EGFP gene was cloned into pCMVori and the control plasmid pcDNA3.1. The two recombinant vectors were transfected into PK-15 cell, and the plasmid DNA and RNA were extracted from the transfected cells. Real-time PCR was used to determine the plasmid replication efficiency of the two plasmids using plasmid before and after Bcl Ⅰ digestion as templates, and the transcription level of the Rep gene in PCV2 replicon was detected by RT-PCR. The average fluorescence intensity of cells transfected with the two plasmids was analyzed with software Image J, and the transcription level of EGFP was determined by means of real-time RT-PCR. The results showed that the replication efficiency of pCMVori in PK-15 cells incubated for 48 h was 136%, and the transcriptions of Rep and Rep' were verified by RT-PCR. The average fluorescence intensity of the cells transfected with pCMVori-EGFP was 39.14% higher than that of pcDNA3.1-EGFP, and the transcription level of EGFP in the former was also 40% higher than that in the latter. In conclusion, the DNA vaccine vector pCMVori constructed in this study can independently replicate in eukaryotic cells. As a result, the expression level of cloned target gene was elevated, providing a basis for developing the pCMVori-based DNA vaccine.
Animals ; Swine ; Circovirus/genetics* ; Vaccines, DNA/genetics* ; Replicon/genetics* ; Genetic Vectors/genetics* ; Plasmids/genetics*

Objective:To explore the survival prognosis of hepatocellular carcinoma patients over 60 years old with comorbidities treated with microwave ablation.Methods:A retrospective analysis of 267 patients with hepatocellular carcinoma aged 60 years or older admitted to the PLA General Hospital from April 2012 to September 2022 were analyzed, including 179 patients with preoperative comorbidities and 88 patients without comorbidities. The overall survival (OS) and progression-free survival (PFS) of the two groups were compared by the Log-rank test, and the Cox proportional hazards regression model was used to evaluate ablation-related risk factors.Results:A total of 267 patients were included (comorbidity group, n=179; no comorbidity group, n=88). There were no statistical differences in OS and PFS between the two groups (all P>0.05). Multivariate Cox regression analysis showed that comorbidities were not risk factors that affected the survival prognosis (OS and PFS) of patients with hepatocellular carcinoma after microwave ablation ( P>0.05). Total bilirubin (hazard ratio 0.356, 95% CI=0.174-0.731, P=0.005) was a risk factor affecting OS; tumor number (hazard ratio 0.538, 95% CI=0.365-0.793, P=0.002) and international coagulation normalized ratio (hazard ratio 1.022, 95% CI=1.001-1.043, P=0.040) were risk factors affecting PFS. Subgroup analysis showed that patients with a maximum diameter of >3 cm and female patients, the OS of the comorbidity group was significantly lower than that of the non-comorbidity group( P<0.05). Conclusions:Microwave ablation therapy remains an effective treatment modality in hepatocellular carcinoma patients over 60 years of age with comorbidities, and its survival prognosis is not inferior to patients with hepatocellular carcinoma without comorbidities.