Aim To investigate the proliferative effect and the apoptosis of rat epididymal epithelial cells induced by podophyllotoxin and its underlying mechanisms.Methods Primary epididymal epithelial cells were cultured in vitro.CCK-8 assay was used to analyze proliferation of epididymal epithelial cells induced by podophyllotoxin on 24, 48 and 72 h.The ultra structural changes of the epididymal epithelial cells were observed by transmission electron microscope.AnnexinV-FITC/PI staining was used to quantify the percentages of apoptosis in the total cell population.The TdTmediated dUTP nick end labeling(TUNEL) technique was applied to observe the morphological changes of apoptotic cells.The expression of tumor necrosis factor alpha(TNF-α) mRNA was investigated by real-time RT-PCR.The level of TNF-α in cell culture supernatant was measured by enzyme-linked immunosorbent assay(ELISA) technology.Western blot was per-formed to determine the protein expression of cytochrome C, caspase-3, caspase-8, caspase-9.Results Podophyllotoxin significantly inhibited the activity of proliferation and induced apoptosis of epididymal epithelial cells in a dose-and time-dependent manner(P<0.05), with a 50％ inhibitory concentration(IC50) value and corresponding 95％ confidence intervals(CI) of 59.36(15.50～227.41), 0.37(0.080～1.70), 0.077(0.017～0.35) μmol·L-1 at 24, 48 and 72 h, respectively.Podophyllotoxin induced cell volume turned round and cell nuclear fragmentation and mitochondrial vacuolation.RT-PCR and ELISA results showed that podophyllotoxin improved the expression of TNF-α mRNA and protein.Western blot results demonstrated that podophyllotoxin activated the protein expression of cytochrome C, caspase-3, caspase-8 and caspase-9.Conclusion Podophyllotoxin can induce rat epididymal epithelial cell apoptosis through both the mitochondria-regulated intrinsic pathway and the TNF receptor-mediated extrinsic pathway.

Objective To continue to study if there is any other pathogenic gene expression related to Th1/Th2 abnormal differentiation,based on the author′s previous results,which have shown that Th1/Th2 unbalance is due to the cytokines and cytokine receptors of differentiation.Methods TaqMan Real time PCR was used to detect the gene expression of Th1/Th2 control in recent onset systemic lupus erythematosus (SLE) patients ( n =38).The genes include I?B,IRF 1,STAT4,GATA3,IL 4R and the others such as CCR1,CCR2,CCR4,CCR5,caepase 1 and CD38,which participate in inflammation,cell apoptosis and so on.Rheumatoid arthritis (RA) patients ( n =50) and normal people ( n =28) were control groups.Results ① Resent onset SLE patients comparing to normal people:STAT4 expression in IL 2/IL 12R ? 2/STAT4 access which induced Th1 differentiation increased significantly ( P 0 05) ;GATA3 expression which induced Th2 differentiation in IL 4/IL 4R/GATA3 access decreased significantly ( P

A capillary electrophoretic method was developed for the separation of enantiomers of naphthylgly-cidic ether(NGE). Several cyclodextrins(CDs) were applied as the chiral selectors and it was found that the ionic modified highly sulfated cyclodextrin(HS-β-CD) could give satisfactory enantioselectivity. In addition,the effects of the pH value of the buffer system,the concentration of the HS-β-CD,capillary temperature and the running voltage on the chiral separation were investigated systematically. The result showed that under the optimized conditions of pH 2. 5,20 mmol/L H_3PO_4-triethanolamine buffer containing 2% highly sulfated cyclodextrin ( HS-β-CD) at capillary temperature 20℃ in the reversed polarity voltage of - 18 kV,the enantiomers of naphthylglycidic ether were baseline separated. This method is simple,precise and can be applied to the chiral separation of naphthylglycidic ether enantiomers and determination of enantiomers excess(ee,% ).

Objectives:To study efficiency characteristics and changes of county hospitals based on data envel-opment analysis model from 2008 to 2012 . Methods:A three-stage DEA model which can exclude the external envi-ronment variable on its efficiency is also added along with the C2 R and BC2-DEA traditional model. Results: From 2008 to 2012, the technical efficiency value was ranging between 0. 723 and 0. 681 in county hospitals, the pure technical efficiency was 0 . 785~0 . 771 and the scale efficiency value scaled between 0 . 908 and 0 . 897 . These values showed a low efficiency, but had annually improved. This reflects that the new health care reform policies have played an important role in improving county hospitals efficiency. Development of medical service ability is behind the development of hospital scale. How to improve the service and management capabilities has gradually become a major direction to improve the county hospital efficiency.

Objective To explore the effect on traditional experiment and case teaching method in regional anatomy study. Methods 80 students from 2014 medical students were randomly selected as the teaching subjects and divided into traditional group and case teaching group. The traditional group con-tained 40 students, using the traditional teaching method, while case teaching group had also 40 students with case teaching method. In the process of teaching, three clinical cases were introduced, including thesubtotal thyroidectomy thoracic outlet syndrome andpancreatic cancer. After the end of the course, the students conducted a unified questionnaire and examination. SPSS 18.0 was used for data line t test or chi square test between the two groups. Results The scores of the students in the case group in the selection questions, blanks and essay questions in the final exam were higher than those of the traditional group; The average total score of the case group was (85.69 ±11.61), while the traditional group was (73.19 ±18.66), and the difference was statistically significant (t=3.597, P=0.002). The results of the questionnaire showed that the students in the case group were higher than the traditional group, and the difference was statistically significant ( χ2=14.753, P=0.001). Conclusion The effect on regional anatomy study with case teaching method is better than the traditional teaching method, and it is a promising teaching reform for the med-ical students.

Objective To explore the pathogenic genes relevant to Behcet's disease (BD) by building the differentail gene expression profiles of peripheral blood leukocytes in BD. Methods Oligonucleotide gene array from Affymetrix Company was applied to study the differed expression levels of whole genome between three age and sex matched BD patients and normal controls. Four genes, BCL6, LRAP, ICOSLG and MME, were selected to be tested for gene expression levels by real-time PCR in the groups of BD, normol controls (NC), Lupus and rheumatoid arthritis (RA) peticnts. Results ① Differential gene expression profile of BD compared to that of normal controls was built up. It contained 89 up-regulated and 57 down-regulated genes. ② Four genes mentioned above had significantly higher expression levels in active BD patients than those in NC but had lower exoression levels in stable BD patients. The expression levels of BCL6 and MME were also proved to be increased significantly in BD than in RA and SLE patients. Conclusion ① Our work shed some light on further research of the etiopathogenesis of BD. ② The expression levels of the four genes are proved to be relevant to BD the first time by us. Further analysis showes that TNF-α and IFN-γ can up-regulate the expression levels of BCL6, LRAP and ICOSLG which may be novel to BD. The MME gene is expressed on the surface of cells, which is convenient for test and may potentially be a marker for the diagnosis of BD.

ObjectiveTo optimize human Hexastatin gene,to express,purify protein and conduct activity experimental research,and to provide a theoretical basis for further study of Hexastatin.MethodsHuman Hexastatin gene was optimized and synthesized.It was connected to the pET28a expression vector,induced to express by isopropyl β-D-1-thiogalactopyranoside(IPTG),and optimized induction conditions.After the ultrasonication of bacterial cells and inclusion bodies,the recombinant fusion protein was purified with Ni-NTA chromatographic column,analyzed and identified by SDS-PAGE and Western Blot,and conduct activity experimental research in vitro by MTT.ResultsConstructed production was pET28a-Hexastatin expression plasmid.The human Hexastatin protein was expressed in E.coli BL21 the high level and accounted for 45.1％ of the total bacterial protein.The purification of recombinant protein purified with Ni-NTA chromatographic column reached 90％,and the concentration was 80 μg/ml.Human Hexastatin protein can restrain the growth of C6,MCF-7 and human vascular endothelial cell (HMEC) cells,and inhibition ratio reach to 72.9％±3.6％,48.8％±2.9％,52.7％±2.5％,respectively through MTT test.ConclusionThe optimized human Hexastatin protein was expressed successfully,which confirmed the inhibition to tumour cells.It is a new way for anti-angiogenesis therapy of tumour.

Objective:Based on the perspective of supply and demand balance, to analyze the implementation effects of primary-feature medical services measures in medical alliances.Methods:From July to September 2019, two regions with different levels of economic development were chosen, and from each region one leading hospital and two community health service centers/township hospitals were selected from its medical alliance. A total of 213 medical staff and 703 residents were surveyed, to learn their awareness and performance appraisal of these feature services.Frequency and proportion were calculated in a descriptive statistical analysis. The scores of the performance appraisal of both supply and demand sides were tested by independent sample t. Results:In the performance appraisal of such measures, the average scoring by medical staff was 4.39±0.07, and that by residents was 3.85±0.06; the residents were more concerned with service capabilities and contents, while medical staff were more concerned with service content and delivery mode; supply and demand sides were found with consensus in terms of improving the medical service quality of chronic diseases in primary level and service accessibility.Conclusions:The county-level medical alliances should focus on enhancing their capacity of primary medical services, and improving their primary service mode with the help of medical insurance institutions. Meanwhile, it is imperative to optimize the primary medical service policy in view of supply and demand balance.

Objective To scientifically measure the workload of basic public health services project on village level .Methods 257 village doctors and 24 public health administrators that work in rural health clinics in Qianjiang city ,Hubei province and Qianjiang distrcit ,Chongqing city were sampled ,with questionnaire survey and key informant in-depth interview methods used to screen basic public health service projects affordable on village level ,for workload measurement .Results 35 basic public health service projects were chosen as the measurement projects regarding their relative workload. Hypertension follow-up workload 100 was set as the benchmark ,finding the rest relative workload of other projects range from11～936 .Conclusions It is reasonable to use the relative value measurement based on RBRVS theory on village level basic public health service projects. Results of this study can offer support for the pricing and further dynamic modification of the basic public health service projects .

Objective:To investigate the clinical and laboratory characteristics of systemic sclerosis (SSc) patients at different age of onset.Methods:Data of SSc patients with onset age ≥18 years old who were registered in the Peking Union Medical College Hospital and Chinese Rheumatism Data Center from August 2008 to June 2020 were included. Patients were divided into 3 groups by the age of onset according to the age segmentation of the World Health Organization. Counting variables were presented as frequency (percentage). Quantitative results were presented as mean±standard deviation, or median, inter quartile range. Differences between groups were analyzed by analysis of variance, the Mann-Whitney test or the chi-square test, depen-ding on the distribution of the variables.Results:Six hundred and eighty-two SSc patients were included. Accor-ding to the age of onset, they were divided into three groups: youth group (18-44 years old), middle-aged group (45-59 years old) and elderly group (over 60 years old). There were 361 patients in the youth group,245 patients in the middle age group and 76 patients in the elderly group. The mean age of onset was (43.8±12.1) years. The variables with significant different among the groups were as the following: left ventricular diastolic dysfunction [14.0%(14/100), 38.8%(39/98), 65.4%(17/26); χ2=30.756, P<0.001]; cardiac arrhythmias [1.9% (7/361), 3.7% (9/361), 7.9% (6/76), χ2=7.38, P=0.024), Raynaud's phenomenon [94.7% (342/361), 89.4%(219/245), 89.5%(68/76), χ2=6.73, P=0.035], loss of finger pad substance [36.9%(133/360), 25.4% (62/244), 18.4% (14/76), χ2=15.184, P=0.001]; digital ulcer [31.0% (112/361), 23.0% (56/244), 15.8% (12/76), χ2=9.86, P=0.007]; arthritis [16.3%(59/361), 13.5%(33/245), 5.3%(4/76), χ2=6.49, P=0.039], digital contracture [11.6%(42/361), 5.7%(14/245), 9.2%(7/76), χ2=6.10, P=0.047]; positive anti-RNP antibody [32.3% (116/359), 20.7% (50/241), 17.3% (13/75), χ2=14.06, P=0.001]; and positive anti-centromere antibody [8.9% (32/351), 18.4%(45/239), 23.7%(18/76), χ2=17.78, P<0.001] were significantly different between the young age group and elder group. Conclusion:The predominant age of disease onset of SSc is middle and young age. Elder onset SSc patients are more likely to have left ventricular diastolic dysfunction, and young onset patients are more likely to have microvascular lesions, which needs more attentions by clinicians.