Objective To optimize the method of primary culture for human amniotic epithelial cells (AECs) and detect the expression of hepatocelluar specific proteins in AECs. Methods AECs were obtained by collagenase and trypsin digestion method. 10,20,40 ng/mL epidermal growth factor (EGF) and 10 ng/mL basic fibroblast growth factor were added separately to the culture media,inverted microscope was used to observe the cell growth and proliferation,and the optimal condition for primary culture of AECs was obtained. The primary cells cultured without growth factors were served as controls. The expression of hepatocelluar specific proteins such as albumin,cytokeratin-18 (CK-18),alpha-1 antitrypsin (AAT) and alpha-1 foetoprotein (AFP) in cultured cells and histological sections were detected by immunohistochemical staining. Results The AECs available were relatively pure,with only a few mesenchymal cells. EGF of 10 ng/mL was chosen as an ingredient of culture medium as the growth and proliferation of AECs significantly accelerated with 10 ng/mL EGF. The expression of albumin,CK-18,AAT and AFP was detected in amnion tissues and AECs cultured in vitro. Conclusion Collagenase and trypsin digestion method and culture medium with 10 ng/mL EGF are favourable conditions for primary culture of AECs. Hepatocelluar specific proteins are expressed in human amnion tissues and AECs cultured in vitro,indicating that AECs have some characteristics of hepatocytes.

Adrenal Hyperplasia, Congenital ; diagnosis ; Female ; Humans ; Infant, Newborn ; Male ; Neonatology ; Pregnancy ; Prenatal Diagnosis

Objective To explore the effect of reducing sub-clinical infections of Toxoplasma gondii in newborn babies with preventive treatment. Methods Forty-four infected pregnant women were treated in different period and their umbilical blood and/or placenta of their newborn babies were tested for IgG, IgM, cAg and DNA of Toxoplasma gondii. Results The positive rate of Toxoplasma DNA in umbilical blood was 37.8%(14/37). The lowest rate was found in the preventive treatment group with a significant difference compared with the group of general treatment. Conclusion Preventive treatment can reduce sub-clinical infection rate of newborn babies effectively.

Objective:To explore the seizure recurrence and prognosis of epilepsy in relapse after anti-epileptic drugs (AEDs) withdrawal, and the influencing factors for these conditions.Methods:From December 2009 to August 2018, patients from the Affiliated Brain Hospital of Nanjing Medical University who relapsed after AEDs withdrawal were collected and followed up for at least 18 months. The seizure recurrence and prognosis of these patients were prospectively observed. The Kaplan-Meier method was used for survival analysis. The associated risk factors of the second relapse in the enrolled patients were analyzed by multivariate Cox analysis. The included patients were divided into good prognosis group and poor prognosis group according to whether they had achieved seizure freedom for at least one year after the first relapse. A multivariate Cox regression model was used to analyze the independent risk factors affecting their prognosis.Results:A total of 56 patients with epilepsy in relapse after AEDS withdrawal were collected. The average follow-up period was 46.23 months (18-120 months) from the initial time of seizure recurrence, and 21 patients (37.5%) had the second seizure recurrence. The relapsing risk in patients who continued to be observed without adding AEDs was higher than those who were treated immediately with drugs [9/16 vs 30.0% (12/40)], but without statistically significant difference (χ2=2.220, P=0.071). The results of univariate analysis showed that focal seizures, seizure frequency more than once per month before remission and poly-drug therapy before AEDs withdrawal were associated with high risk of the second relapse. Poly-drug therapy was an independent risk factor for the second relapse by multivariate Cox analysis ( HR=3.383, 95% CI 1.257-9.105). Of the 56 patients with epilepsy in relapse after AEDs withdrawal, 47 patients (83.9%) had a good prognosis without seizure for at least one year, and of 33 patients who were followed up for three years or more, 26 (78.8%) had no seizure for at least two years. Between the group retreated immediately after the first recurrence and the group without immediate treatment [87.5% (35/40) vs 12/16],there were no statistically significant differences on the proportions of good prognosis (χ2=2.333, P=0.258). Univariate analysis showed that the course of epilepsy>6 months before initial treatment, the frequency of seizures>1/month before remission, symptomatic epilepsy and poly-drug therapy were associated with the poor prognosis. However, none of independent risk factors was found for the poor prognosis through the multivariate analysis. Conclusions:The prognosis of patients with epilepsy in relapse after AEDs withdrawal is well, and about 2/3 patients with epilepsy in relapse after AEDs withdrawal have no more seizure recurrences. The poly-drug therapy before AEDs withdrawal may be an independent risk factor for the second seizure relapse.

Objective To discuss the accuracy and objectivity of student standardized patients (SSP) in objective structured clinical examination(OSCE). Methods On March 30 and 31, 2013, 168 seven-year program medical students of class 2006 and 2007 took part in OSCE. Differences in as-sessment results between SPP and those given by professional doctors at 3 SP sites(angina pectoris SSP station, acute cholecystitis SSP station, depression SSP station) were analyzed. Each site had 4 items for assessment, with a total score of 100. Scores were given in strict accordance with a set of unified scoring rules. Counting data were presented as x±s. Data were verified using t test. P<0.05 was considered statis-tically considerable. Results At angina pectoris station, respective scores of SSP and professional physi-cians were 85.2±7.1 and 85.5±6.6, P=0.688. At acute cholecystitis station, respective score of SSP and professional physicians were 89.1±5.2 and 88.2±6.2, P=0.150. At depression station, respec-tive score of SSP and professional physicians were 79.8±7.5 and 78.2±7.0, P=0.078. Conclusion There is no statistical difference between scores given by SSP and those given by the physicians in OSCE. This proves that SSP who received standardized training delivers fair and accurate results in OSCE , and therefore is recommended for future application.

Objective To explore the value of shear wave elastrography (SWE) in diagnosis of prostate cancer.Methods SWE quantitative elastography was preformed in 55 patients with suspected prostate cancer,to obtain the elastic modulus (mean,maximum).Each patient underwent sonography-guided prostatic biopsy on the same day.With the pathologic results as reference,ROC curves were used to assess diagnostic performance.Results ①Pathological tests showed that 39 lesions were benign(hyperplasia) and 16 lesions were malignant.The mean elasticity value of benign lesions was (39.04 ± 8.22) kPa,and the maximum value was (54.10 ± 9.18)kPa,whereas of malignant ones were (53.31 ± 3.92)kPa and (68.71 ±2.57)kPa,respectively (P ＜0.05).② The area under the ROC curve (AZ) of the maximum and mean elasticity value was 0.951 and 0.944.Taking 48.07 kPa as the threshold of the mean elasticity value,the sensitivity was 91.8％ and the specificity was 89.7％.Then taking 65.50 kPa as the threshold of the maximum elasticity value,the sensitivity was 92.1％ and the specificity was 87.5％.Conclusions SWE is helpful to diagnose and differentiate prostate diseases by measuring elastic modulus.

Objective To investigate drug resistance mechanism of aminoglycoside-resistant Acinetobacter baumannii by detecting 16S rRNA methylase gene and three common genes of aminoglycoside-modifying enzymes in Acinetobacter baumannii infected patients at EICU. Methods The 48 Acinetobacter baumannii strains were collected,and antimicrobial susceptibility tests were performed by VITEK automicroscan. The MIC was detected by 2-fold agar dilution method,and genes were analyzed by polymerase chain reaction( PCR) . Results Among 48 strains,28 were highly resistant to aminoglycosides and 20 showed lower resistances. The 16S rRNA armA,APH(3')-I,ANT(3'')-Ia,AAC(6')-Ib genes were detected in 71. 43%,60. 71%,82. 14%, and 53. 57%of the 28 highly resistant strains,but only present in 0. 00%,0. 05%,0. 05%,and 0. 05%of the low-resistant isolates(P<0. 01). Conclusion The aminoglycoside-modifying enzymes and 16S rRNA methylase were frequently found in Acinetobacter baumannii clinical isolates,which is closely related to the high-level resistance to aminoglycoside antibiotics.

Objective To investigate the effect of recombinant adenovirus mediatied human endostatin (rAD-GFP-ES)on rats with collagen typeⅡinduced arthritis(CIA),and explore the mechanism of inflamma- tion and cytokines inhibition on rats CIA.Methods The rAD-GFP-ES was amplified and purified.The model of rat CIA was induced by intradermal injection of typeⅡcollagen combined with complete Freund's adjuvant(CFA). On the second day after the injection,the therapeutic administration of rAD-GFP-ES(1?10~(11)pfu?kg~(-1)?week~(-1)?4 weeks)were performed to the rats.The mean arthritis index(AI)was scored every week since then.The relative concentrations of ES,IL-I?,TNF-?in sera collected at the fourth week were evaluated by western blotting. Results①The titer of the purified rAD-GFP-ES and rAD-GFP was 6.6?10~(12)pfu/ml and 4.8?10~(12)pfu/ml,re- spectively(A_(260nm)/A_(280nm)＞1.3).②The concentration of ES in sera of the group treated with rAD-GFP-ES was 2.4-lold higher compared to the normal group.③The mean arthritis index of the group treated with rAD-GFP- ES was much lower than that of the model group.The administration of rAD-GFP-ES could significantly de- creas the production of IL-1?and TNF-?in sera.Conclusions①The rAD-GFP-ES is efficiently expressed in vivo.②The rAD-GFP-ES has an inhibitory effect on the arthritis index of rat CIA.③IL-1?and TNF-?are involved in the pathogenesis of RA.The rAD-GFP-ES has an inhibitory effect on the expression of IL-1?and TNF-?in rat CIA.

Objective: To examine the associations of fibroblast growth factor 19 (FGF19), its co-receptor KLB gene and its receptor FGFR4 with susceptibility to sarcopenia, so as to provide insights into elucidation of sarcopenia pathogenesis and formulation of precision interventions for sarcopenia. Methods: A case-control study was conducted. Patients with sarcopenia at ages of 60 years and older included in the Zhejiang Provincial Elderly Health Surveillance Cohorts were selected as the sarcopenia group, and normal residents at ages of 60 years and older were served as controls. Subjects' demographics were collected using questionnaire surveys, and the height, body weight, appendicular skeletal muscle mass and grip strength were measured. Genomic DNA was extracted from blood samples for multiplex PCR targeted capture. The associations between the KLB gene single-nucleotide polymorphisms (SNPs) and susceptibility to sarcopenia were evaluated using multivariable logistic regression models. Results: There were 200 cases in the sarcopenia group, including 91 men and 109 women, and 180 cases in the control group, including 70 men and 110 women. All SNPs satisfied the Hardy-Weinberg equilibrium, and the minor allele frequencies were all > 0.05. There were no significant differences in the distribution of SNPs between the sarcopenia and control groups (all P>0.05). Multivariable logistic regression analysis showed that the SNP rs2687968 locus in the KLB gene was significantly associated with the susceptibility to sarcopenia among the elderly men (superdominant model), and individuals carrying the AC allele had a 2.332-fold higher risk of sarcopenia than those carrying the AA/CC allele (95%CI: 1.882-3.313). Conclusions KLB gene may correlate with the susceptibility to sarcopenia among the elderly men.

Objective:To compare the survival and prognostic factors of intraoperative radiotherapy (IORT) and postoperative radiotherapy (PORT) in female patients, aged≥50 years, diagnosed with node-negative breast cancer (≤ 3 cm in size).Methods:Clinical data of eligible early breast cancer patients between 2010 and 2015 were obtained from the SEER database. Patients were divided into the IORT and PORT groups according to the radiotherapy record and propensity score matching (PSM) was subsequently conducted. Kaplan-Meier curve was used to evaluate the overall survival (OS) and breast cancer-specific survival (BCSS) between two groups and Cox proportional hazard regression analysis was used to explore the risk factors of clinical prognosis.Results:7 068 patients were included after PSM. The median follow-up time was 32.0 months. The 5-year OS rates in the IORT and PORT groups were 96.8% and 93.8%, respectively. Univariate Cox analysis showed that radiotherapy, age, histological grade, T stage, estrogen receptor (ER) status and progesterone receptor (PR) status were the independent risk factors for OS, and histological grade, T stage, ER status, PR status and chemotherapy were the independent risk factors for BCSS. Multivariate Cox regression analysis demonstrated that patients who received IORT had better OS than PORT counterparts ( P=0.020). Besides, patients aged≥60 years obtained worse OS than those aged<60 years ( P=0.003). Patients with T 2 stage or ER-negative tumors had worse OS than those with T 1 stage tumors ( P<0.001) or ER-positive tumors ( P=0.001). Patients with grade Ⅲ-Ⅳ tumors achieved worse BCSS ( P=0.004). Subgroup analysis showed that IORT yielded better OS for elderly patients (≥60 years), grade Ⅲ-Ⅳ tumors, infiltrating duct carcinoma, T 2 stage tumors, ER-positive tumors, PR-positive tumors and patients without chemotherapy. Conclusions:IORT may bring benefit for highly selected patients with low risk of recurrence, which is not inferior to PORT in terms of short-term survival. Prospective studies with longer follow-up time are needed to confirm the findings.