Objective:To introduce a "Four-step method" for extraction and separation of Candida albicans total proteins.Methods: Proteins of C.albicans were extracted step-by-step with 4 kinds of solutions with different solubilities.After quantification,the protein samples were separated by isoelectric focusing electrophoresis and then by SDS-PAGE.Results: Proteins with different solubilities were successfully extracted step-by-step from C.albicans and were separated by two-dimensional gel electrophoresis. Conclusion: The "Four-step method" for extraction of C.albicans proteins is an effective approach to study C.albicans membrane proteome and lays a foundation for further investigation of the mechanisms of antifungal agents and drug resistance in C.albicans.

The Y118A、Y118F、Y118T、S378A、S378T、H310A、H310R mutants of Candida albicans sterol 14?-demethylase (CACYP51) were constructed and heterologously expressed in D12667, the reconstructed strain with the deletion of CYP51 gene of the Y12667. With the strains obtained and microsome enzymes separated, the western blot and the ultraviolet absorption spectrophotometry were used to qualitative and quantitative detect the expressed protein, the GC-MS was used to detect the metabolism activity of the protein. The results showed that, the target protein expressed successfully in the reconstructed strains, with the expression level up to 25% of the total microsome proteins. The results also showed that, the wild type protein had the catalytic activity to its nature substrate. While after alteration the wild gene with Y118A、Y118F、Y118T、S378A、S378T、H310A、H310R by a single base substitution, the catalytic activity of protein markedly decreased respectively. So the wild type and mutation CYP51 were expressed successfully in Saccharomyces cerevisiae and the expression products preserved the activity to metabolism their nature substrate.

The promoter of mitochondria-localized small heat shock protein gene in Lycopersicon esculentum (Lehsp23.8) is characterized as strongly heat-inducible. In this study, to determine how the expression of Lehsp23.8 is regulated, we conducted five expression vectors carrying the gus gene driven by the 5' deletion products of the Lehsp23.8 promoter. The corresponding transgenic tobacco plants were generated via Agrobacterium tumefaciens-mediated transformation. Transgenic plants were identified by PCR and Southern blotting analysis. GUS activities under heat-shock conditions were characterized in transgenic tobacco plants. After heat shock, obvious GUS staining was detected in the leaves, shoots, roots, flowers and fruits of the transgenic tobacco plants. The result of fluorometric GUS assays in leaves showed that the heat-induced GUS activity of the 565 bp promoter was the strongest, while that of the 255 bp promoter was the lowest. Deletion analysis shows that the smallest promoter fragment (-255 bp to -23 bp) is sufficient for heat induction. It also indicates that the sequences between -255 bp and -565 bp serve as enhancers, while the sequences between -565 bp and -871 bp can repress the heat-induced activity of the Lehsp23.8 promoter.
Gene Expression Regulation, Plant ; genetics ; Genetic Vectors ; Heat-Shock Proteins ; genetics ; Heat-Shock Proteins, Small ; genetics ; metabolism ; Hot Temperature ; Lycopersicon esculentum ; genetics ; Mitochondria ; genetics ; metabolism ; Plants, Genetically Modified ; genetics ; Promoter Regions, Genetic ; genetics ; Tobacco ; genetics

Hepatotoxicity is one of the most common adverse reactions during anti -hyperlipidemia treatment .Mechanisms of anti-hyperlipidemia drug-induced hepatotoxicity are not clear yet , but most of the toxic reactions are dose-related hypersensitivity and could be released after drug withdrawal .It is accepted that clinical risk factors for the development of hepatotoxicity during anti -hyper-lipidemia treatment are high age and chronic illnesses .Treatment of anti-hyperlipidemia drug-induced hepatotoxicity has not been uni-fied and most of the treatments are non-specific and symptomatic .Researching hypotoxicity and hepatoprotection antihyperlipidemia drug, especially TCM and pharmaceutics will become a promising direction .

uate and should be standardized.

Four cases of Pentalogy of Cantrell were diagnosed by prenatal ultrasonography and confirmed by autopsy after labor induction.There were 1 case of complete Pentalogy of Cantrell and 3 cases of incomplete Pentalogy.In 3 cases of incomplete Pentalogy of Cantrell,1 case presented with high umbilical cord prolapse,extraspectoral heart,partial defect of the lower part of the sternum,partial defect of the pericardium,single umbilical artery and cervical lymphatic cyst;1 case presented with high umbilical cord prolapse,extraspectoral heart,thoracic fissure and ventricular septal defect;and 1 case presented with umbilical cord,extraspectoral heart,partial inferior sternal segment defect,ventricular septal defect and bilateral clubfoot varus.In l case of complete Pentalogy of Cantrell,there were high umbilical cord bulge,extraspectoral heart,chest fissure and inferior segment defect,anterior diaphragm and pericardium defect,total forebrain deformity,right 12th ribs loss and ventricular septal defect.Ectopic heart and omphalocele are typical echographic features of Pentalogy of Cantrel and prenatal ultrasonography is of important value in early diagnosis of this congenital malformation.

Objective:To understand the 10-year risk of ischemic cardiovascular disease (ICVD) in hypertensive patients in communities in Shenzhen, and to provide baseline data for the comprehensive implementation of community-based hypertension health management measures based on ICVD risk assessment.Methods:From July 2017 to June 2018, 200 community health service centers in 10 districts of Shenzhen were selected by multi-stage random sampling to investigate hypertensive patients in communities aged≥35. After the patients who met the inclusion criteria signed the informed consent form, the demographic characteristics, physical examination, and laboratory tests were conducted. The absolute risk of ICVD in 10 years was evaluated by using the ICVD risk assessment module from the Shenzhen community health service information system. According to the China 10-year risk for ICVD score tables, recommended by the Chinese cardiovascular disease prevention guidelines, the risk grade of ICVD was estimated and analyzed. In addition, the relative risk was used to eliminate the influence of gender and age, and the single sample t-test was used for comparisons with the larger population. Results:30 015 hypertensive patients from communities were investigated, including 16 165 (53.86%) males and 13 850 (46.14%) females, aged 35-102 years, with a median age of 59.0. 6 405 (21.34%), 5 771 (19.23%) and 17 839 (59.43%) were at high, medium and low risk of ICVD, respectively. The absolute risk was 3.77%, 95% CI (3.72%-3.82%). The risk grades of ICVD in 10 years was different as per gender, age, marital status, population type， smoking, family history of hypertension, history of diabetes mellitus, systolic blood pressure, body mass index, and total cholesterol level, P values were all<0.001. The risk grades of ICVD in males, married patients, floating populations, having a total cholesterol <5.20 mmol/L, non-smokers, non-diabetics, and having a family history of hypertension were lower than that of other characteristics, and increased with age, systolic blood pressure, and body mass index. Further analysis of 15 652 patients aged 35-59 years old showed that the 10-year risk of ICVD in patients with hypertension in communities was slightly higher than that of the larger population, and the relative risk was 1.27, 95% CI (1.25-1.28), t=37.648, P<0.001. In males, relative risk was 0.94, 95% CI (0.93-0.95), t= 9.906, P<0.001, and there were no significant trend changes among age groups. In females, relative risk was 2.05, 95% CI (2.01-2.09), t=72.207, P<0.001, and increased with age. Conclusion:The 10-year risk of ICVD in hypertensive patients is often at a high level in Shenzhen, so it is necessary to carry out the risk assessment of ICVD in the community. The effect of community hypertension health management on reducing the risk of ICVD may vary between genders. The risk of ICVD in male patients is lower than that of males in the larger population, but in female patients it is still much higher.

Objective : To study the protective effect of polydatin on lipopolysaccharide-induced injury of human umbilical vein vascular endothelial cells (HUVECs) through the protein Janus kinase 2 (JAK2) / signal transducers and activators of transcription 3 ( STAT3) signaling pathway． Methods : HUVECs were cultured in vitro，and 500 ng / ml LPS induced their injury and set as a model group ; based on the model group，endothelial cells were intervened with different concentrations ( 10，20，and 40 μmol / L ) of polydatin for 24 h，and set as polydatin low concentration group，polydatin medium concentration group，and polydatin high concentration group，respectively ; a control group was set as another group．CCK-8，monocyte-endothelial cell adhesion，scratch and Transwell assays were used to detect cell viability，adhesion，migration and invasive ability ; ELISA was used to detect interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-α) levels in the cell supernatant ; Western blot was used to detect the expression of proteins related to the JAK2 / STAT3 signaling pathway levels of JAK2 / STAT3 signaling pathway relat- ed proteins. Results : Compared with the control group，the model group showed decreased cell survival ( P < 0. 01) ，increased cell adhesion，migration and invasion (P＜0. 001，P ＜0. 05，P ＜0. 01) ，increased levels of IL-6 and TNF-α in the cell supernatant (P＜0. 001) ，and increased levels of phosphorylation of JAK2 and STAT3 pro- teins in the cells (P＜0. 05) ．Compared with the model group，LPS damage to cells was attenuated after polydatin intervention，cell survival was increased in polydatin low －，medium － and high － concentration groups ( P < 0. 05) ，cell adhesion，migration，and invasion decreased (P＜0. 05，P＜0. 05，P＜0. 001) ，IL-6 and TNF-α levels in cell supernatants decreased (P＜0. 05) ，and the levels of cellular JAK2 and STAT3 protein phosphorylation lev- els decreased (P＜0. 05) ． Conclusion Polydatin seems to reduce the inflammatory injury of human umbilical vein endothelial cells induced by LPS，reducing the secretion of inflammatory factors，and inhibiting the ability of cell ad- hesion，migration and invasion，which may be related to the down-regulation of JAK2 / STAT3 signal pathway by polydatin.

Objective: To understand the possible transmission route of a family cluster of COVID-19 in Zhengzhou and the potential infectivity of COVID-19 in incubation period, and provide scientific evidence for the timely control of infectious source and curb the spread of the epidemic. Methods: Epidemiological investigation was conducted for a family cluster of COVID-19 (8 cases) with descriptive epidemiological method, and respiratory tract samples of the cases were collected for the nucleic acid detection of 2019-nCoV by RT-PCR. Results: Two primary cases, which occurred on 31 January and 1 February, 2020, respectively, had a common exposure history in Wuhan. The other six family members had onsets on 30 January, 31 January, 1 February (three cases) and 3 February, 2020. Conclusions In this family cluster of COVID-19, six family members were infected through common family exposure to the 2 primary cases. Five secondary cases had onsets earlier than or on the same day as the primary cases, indicating that COVID-19 is contagious in incubation period, and the home isolation in the early phase of the epidemic might lead to the risk of family cluster of COVID-19.

Objective:To investigate the epidemiological characteristics, diagnosis, treat-ment and prognosis of gallbladder cancer in China from 2010 to 2017.Methods:The single disease retrospective registration cohort study was conducted. Based on the concept of the real world study, the clinicopathological data, from multicenter retrospective clinical data database of gallbladder cancer of Chinese Research Group of Gallbladder Cancer (CRGGC), of 6 159 patients with gallbladder cancer who were admitted to 42 hospitals from January 2010 to December 2017 were collected. Observation indicators: (1) case resources; (2) age and sex distribution; (3) diagnosis; (4) surgical treatment and prognosis; (5) multimodality therapy and prognosis. The follow-up data of the 42 hospitals were collected and analyzed by the CRGGC. The main outcome indicator was the overall survival time from date of operation for surgical patients or date of diagnosis for non-surgical patients to the end of outcome event or the last follow-up. Measurement data with normal distribu-tion were represented as Mean±SD, and comparison between groups was conducted using the t test. Measurement data with skewed distribution were represented as M( Q1, Q3) or M(range), and com-parison between groups was conducted using the U test. Count data were described as absolute numbers or percentages, and comparison between groups was conducted using the chi-square test. Univariate analysis was performed using the Logistic forced regression model, and variables with P<0.1 in the univariate analysis were included for multivariate analysis. Multivariate analysis was performed using the Logistic stepwise regression model. The life table method was used to calculate survival rates and the Kaplan-Meier method was used to draw survival curves. Log-rank test was used for survival analysis. Results:(1) Case resources: of the 42 hospitals, there were 35 class A of tertiary hospitals and 7 class B of tertiary hospitals, 16 hospitals with high admission of gallbladder cancer and 26 hospitals with low admission of gallbladder cancer, respectively. Geographical distribution of the 42 hospitals: there were 9 hospitals in central China, 5 hospitals in northeast China, 22 hospitals in eastern China and 6 hospitals in western China. Geographical distribution of the 6 159 patients: there were 2 154 cases(34.973%) from central China, 705 cases(11.447%) from northeast China, 1 969 cases(31.969%) from eastern China and 1 331 cases(21.611%) from western China. The total average number of cases undergoing diagnosis and treatment in hospitals of the 6 159 patients was 18.3±4.5 per year, in which the average number of cases undergoing diagnosis and treatment in hospitals of 4 974 patients(80.760%) from hospitals with high admission of gallbladder cancer was 38.8±8.9 per year and the average number of cases undergoing diagnosis and treatment in hospitals of 1 185 patients(19.240%) from hospitals with low admission of gallbladder cancer was 5.7±1.9 per year. (2) Age and sex distribution: the age of 6 159 patients diagnosed as gallbladder cancer was 64(56,71) years, in which the age of 2 247 male patients(36.483%) diagnosed as gallbladder cancer was 64(58,71)years and the age of 3 912 female patients(63.517%) diagnosed as gallbladder cancer was 63(55,71)years. The sex ratio of female to male was 1.74:1. Of 6 159 patients, 3 886 cases(63.095%) were diagnosed as gallbladder cancer at 56 to 75 years old. There was a significant difference on age at diagnosis between male and female patients ( Z=-3.99, P<0.001). (3) Diagnosis: of 6 159 patients, 2 503 cases(40.640%) were initially diagnosed as gallbladder cancer and 3 656 cases(59.360%) were initially diagnosed as non-gallbladder cancer. There were 2 110 patients(34.259%) not undergoing surgical treatment, of which 200 cases(9.479%) were initially diagnosed as gallbladder cancer and 1 910 cases(90.521%) were initially diagnosed as non-gallbladder cancer. There were 4 049 patients(65.741%) undergoing surgical treatment, of which 2 303 cases(56.878%) were initially diagnosed as gallbladder cancer and 1 746 cases(43.122%) were initial diagnosed as non-gallbladder cancer. Of the 1 746 patients who were initially diagnosed as non-gallbladder cancer, there were 774 cases(19.116%) diagnosed as gallbladder cancer during operation and 972 cases(24.006%) diagnosed as gallbladder cancer after operation. Of 6 159 patients, there were 2 521 cases(40.932%), 2 335 cases(37.912%) and 1 114 cases(18.087%) undergoing ultrasound, computed tomography (CT) or magnetic resonance imaging (MRI) examination before initial diagnosis, respec-tively, and there were 3 259 cases(52.914%), 3 172 cases(51.502%) and 4 016 cases(65.205%) undergoing serum carcinoembryonic antigen, CA19-9 or CA125 examination before initially diagnosis, respectively. One patient may underwent multiple examinations. Results of univariate analysis showed that geographical distribution of hospitals (eastern China or western China), age ≥72 years, gallbladder cancer annual admission of hospitals, whether undergoing ultrasound, CT, MRI, serum carcinoembryonic antigen, CA19-9 or CA125 examination before initially diagnosis were related factors influencing initial diagnosis of gallbladder cancer patients ( odds ratio=1.45, 1.98, 0.69, 0.68, 2.43, 0.41, 1.63, 0.41, 0.39, 0.42, 95% confidence interval as 1.21-1.74, 1.64-2.40, 0.59-0.80, 0.60-0.78, 2.19-2.70, 0.37-0.45, 1.43-1.86, 0.37-0.45, 0.35-0.43, 0.38-0.47, P<0.05). Results of multivariate analysis showed that geographical distribution of hospitals (eastern China or western China), sex, age ≥72 years, gallbladder cancer annual admission of hospitals and cases undergoing ultrasound, CT, serum CA19-9 examination before initially diagnosis were indepen-dent influencing factors influencing initial diagnosis of gallbladder cancer patients ( odds ratio=1.36, 1.42, 0.89, 0.67, 1.85, 1.56, 1.57, 0.39, 95% confidence interval as 1.13-1.64, 1.16-1.73, 0.79-0.99, 0.57-0.78, 1.60-2.14, 1.38-1.77, 1.38-1.79, 0.35-0.43, P<0.05). (4) Surgical treatment and prognosis. Of the 4 049 patients undergoing surgical treatment, there were 2 447 cases(60.435%) with complete pathological staging data and follow-up data. Cases with pathological staging as stage 0, stage Ⅰ, stage Ⅱ, stage Ⅲa, stage Ⅲb, stage Ⅳa and stage Ⅳb were 85(3.474%), 201(8.214%), 71(2.902%), 890(36.371%), 382(15.611%), 33(1.348%) and 785(32.080%), respectively. The median follow-up time and median postoperative overall survival time of the 2 447 cases were 55.75 months (95% confidence interval as 52.78-58.35) and 23.46 months (95% confidence interval as 21.23-25.71), respectively. There was a significant difference in the overall survival between cases with pathological staging as stage 0, stage Ⅰ, stage Ⅱ, stage Ⅲa, stage Ⅲb, stage Ⅳa and stage Ⅳb ( χ2=512.47, P<0.001). Of the 4 049 patients undergoing surgical treatment, there were 2 988 cases(73.796%) with resectable tumor, 177 cases(4.371%) with unresectable tumor and 884 cases(21.833%) with tumor unassessable for resectabi-lity. Of the 2 988 cases with resectable tumor, there were 2 036 cases(68.139%) undergoing radical resection, 504 cases(16.867%) undergoing non-radical resection and 448 cases(14.994%) with operation unassessable for curative effect. Of the 2 447 cases with complete pathological staging data and follow-up data who underwent surgical treatment, there were 53 cases(2.166%) with unresectable tumor, 300 cases(12.260%) with resectable tumor and receiving non-radical resection, 1 441 cases(58.888%) with resectable tumor and receiving radical resection, 653 cases(26.686%) with resectable tumor and receiving operation unassessable for curative effect. There were 733 cases not undergoing surgical treatment with complete pathological staging data and follow-up data. There was a significant difference in the overall survival between cases not undergoing surgical treatment, cases undergoing surgical treatment for unresectable tumor, cases undergoing non-radical resection for resectable tumor and cases undergoing radical resection for resectable tumor ( χ2=121.04, P<0.001). (5) Multimodality therapy and prognosis: of 6 159 patients, there were 541 cases(8.784%) under-going postoperative adjuvant chemotherapy and advanced chemotherapy, 76 cases(1.234%) under-going radiotherapy. There were 1 170 advanced gallbladder cancer (pathological staging ≥stage Ⅲa) patients undergoing radical resection, including 126 cases(10.769%) with post-operative adjuvant chemotherapy and 1 044 cases(89.231%) without postoperative adjuvant chemo-therapy. There was no significant difference in the overall survival between cases with post-operative adjuvant chemotherapy and cases without postoperative adjuvant chemotherapy ( χ2=0.23, P=0.629). There were 658 patients with pathological staging as stage Ⅲa who underwent radical resection, including 66 cases(10.030%) with postoperative adjuvant chemotherapy and 592 cases(89.970%) without postoperative adjuvant chemotherapy. There was no significant difference in the overall survival between cases with postoperative adjuvant chemotherapy and cases without postoperative adjuvant chemotherapy ( χ2=0.05, P=0.817). There were 512 patients with pathological staging ≥stage Ⅲb who underwent radical resection, including 60 cases(11.719%) with postoperative adjuvant chemotherapy and 452 cases(88.281%) without postoperative adjuvant chemotherapy. There was no significant difference in the overall survival between cases with postoperative adjuvant chemo-therapy and cases without post-operative adjuvant chemo-therapy ( χ2=1.50, P=0.220). Conclusions:There are more women than men with gallbladder cancer in China and more than half of patients are diagnosed at the age of 56 to 75 years. Cases undergoing ultrasound, CT, serum CA19-9 examination before initial diagnosis are independent influencing factors influencing initial diagnosis of gallbladder cancer patients. Preoperative resectability evaluation can improve the therapy strategy and patient prognosis. Adjuvant chemotherapy for gallbladder cancer is not standardized and in low proportion in China.