The Activating Transcription Factor 2 (ATF2) mainly regulates gene transcriptions through mediating cellular response to environmental stimulus. Accumulating studies show that ATF2 is involved in tumorigenesis and tumor cell proliferation, differentiation , apoptosis , invasion and metastasis. Activation of ATF2 may promote tumor cells growth, however, there are several studies show that ATF2 might inhibit growth of tumor cell. Further understanding of the role for ATF2 in tumorigenesis might provide novel thinking pathways for tumor etiology and therapy.

Objective To establish the specific 16S 23S rRNA gene spacer regions map of different bacteria by PCR, RFLP(restriction fragment length polymorphism ),DNA clone and sequences analysis. Methods A pair of primer was selected from highly conserved sequences adjacent to the 16S 23S rRNA spacer region. The farget rRNA regions from 61 strains of standard bacteria and corresponding clinical isolates representing for 20 genera and 26 species were amplified by PCR,and thereafter analyzed RFLP, DNA clone and sequences analysis.Meanwhile, all the specimens were examined by bacterial culture and PCR RFLP analysis. Results 26 different standard strains presented one band,two bands,three bands and more than three bands respectively, the sensitivity of which reached 2.5 CFU and had no cross reaction to the human genomic DNA,fungus and virus.14 species could be distinguished immediately by PCR, other 10 species must be identified by further Hinf I or Alu I digestion. K.pneumoniae and E.durans differentiate only at the site of 779 th nucleotide according to the sequence analysis, and only one enzyme Xma III could discriminate them.15 specimens from 42 septicemic neonates were blood culture positive and the positive rate was 35.7%. However, 27 specimens were positive by PCR and the positive rate was 64.2%,which was significantly higher than that of the blood culture( P

To report the process of diagnosis and treatment of 1 case with SRY gene mutation of 46, XY complete gonadal dysplasia, and to discuss the clinical characteristics, diagnosis and treatment of the disease.Due to clitoral enlargement for 8 months, a 9 years old girl was admitted to the Children′s Hospital Affiliated to Zhejiang University School of Medicine.Previously, she had early breast development, and suffered from high gonadotropin expression when she was 6 years and 4 months old.Physical examination: breast B3 stage, female vulva, clitoris hypertrophy, normal urethra, normal vaginal opening, slightly thick hymen ring, the development of pubic hair was 2 stages, and Prader score level 1.Laboratory data showed elevated levels of estradiol, testosterone, and human chorionic gonadotrophin.Genetic examination revealed that the chromosome karyotype was 46, XY and SRY gene detection was positive.Therefore, the patient was diagnosed with 46, XY complete gonadal dysplasia.Bilateral gonadectomy was performed, and the posto-perative pathological diagnosis was bilateral gonadoblastoma with left dysgerminoma.The tumor did not recur after che-motherapy.The etiology of early breast development needs to be carefully identified.Patients with sexual characteristics dysplasia need to accept the chromosome karyotype analysis and gene detection, and surgical exploration should be performed when necessary for a correct diagnosis as soon as possible.

Objective To evaluate the value of serum uric acid(UA)levels with reference to the age,waist circumference,and body mass index(BMI)in predicting the metabolic syndrome(MS)in obese children.Methods A total of 300 obese children,including 180 boys and 120 girls,were enrolled in this study.The height,BMI,waist and hip circumference,blood pressure,serum glucose,insulin and lipid profile in all participants were measured.Oral glucose tolerance test and insulin releasing test were performed.The boys or girls were divided into 4 groups according to the 4 quantile of UA level,respectively.The clinical characteristics and correlation of UA with the clinical indexes and MS components were compared.The binary Logistic regression analysis was applied in the risk of MS and its components for the 4 groups of obese children.The area under the receiver operating characteristic curve(ROC curve)of UA level,age,waist circumference and BMI were used to predict the MS.Results UA level was increased with the increase of age,waist circumference and BMI,and the UA level was significantly correlated with triacylglycerol,postprandial 2 h glucose(2 h PG)(r=0.196,0.174 in boys;r=0.291,0.179 in girls).In boys,the adjusted odds ratio and 95％CI of the highest quartile of UA for triglyceridemia was 2.71(95％CI:0.77-9.58);which in girls,the adjusted odds ratio and 95％CI of the highest quartile of UA for hyperglycemia,hypertension were 8.45(95％CI:1.76-40.52)and 3.93(95％CI:0.66-23.33),respectively,with significant differences.In boys,the area under the ROC curve of UA level,age,waist circumference and BMI which predict the MS were 0.652 0.626,0.621,0.62,respectively,and the differences were significant(all P<0.05).Conclusions The UA level is significantly correlated with the composition of MS,UA detection combining with reference to the age,waist circumference,and BMI is helpful for the identification of high risk groups of metabolic syndrome.

Objective: Ovarian carcinosarcoma (OCS) is one of rarest and most challenging histologic subtype of ovarian cancer. It features remarkable cellular heterogeneity. Using single-cell RNA sequencing (scRNA-seq), we characterize the cellular composition of OCS and identify their molecular characteristics. Methods: We applied scRNA-seq to resected primary OCS for the in-depth analysis of tumor cells and the tumor microenvironment. Immunohistochemistry staining was used for validation. Results: Malignant epithelial and fibroblast cells displayed a high-degree of intratumoral heterogeneity. We revealed that certain epithelial cell subclusters had high levels of drug resistance scores and many active metabolic pathways. Furthermore, γδ T cells exhibited enriched interferon (IFN)-γ and IFN-α response characteristics. Analyzing ligand-receptor interaction pairs between cell types, we identified broadly interacting cells and observed an interaction between the ANXA1+ epithelial population and FPR1+/FPR3+ myeloid cells. Conclusion Our findings provide a single-cell transcriptomic signature of human OCS and present a well-established resource for elucidating OCS diversity.

Objective: To investigate and analyze the awareness, utilization, and satisfaction of patients with chronic diseases (hypertension and type 2 diabetes mellitus), as well as the influencing factors related to health management services for chronic diseases in basic public health service projects in Zhejiang Province, in order to promote the development of effective health management services for chronic diseases. Methods: Between September and November 2017, 960 local patients with chronic diseases aged 35 years or older were randomly selected in eight counties (cities/districts) in Zhejiang province using the random number table, based on the chronic disease information from the information management system. A total of 394 (41.04%) male and 566 (58.96%) female patients, with an average age of (68.02±10.02) years, participated in this study. Face-to-face questionnaires were used to collect patients’ awareness, utilization, and satisfaction with health management services for chronic diseases. The awareness of health management services for chronic diseases and their utilization status were described and analyzed by statistical rate indicators. Possible factors affecting the awareness and utilization status were analyzed by χ² test and logistic regression. Satisfaction scores of patients with chronic diseases were analyzed by means and standard deviation. T-test was used to compare the satisfaction level with service items (service convenience, service environment, waiting time, medical skill level, and service attitude) between urban and rural patients with chronic diseases. Finally, multivariate logistic regression analysis was conducted to determine overall satisfaction. Results: The overall awareness of health management services for chronic diseases in chronic disease patients was 96.15%. Multivariate Logistic regression analysis showed that patients who lived in developed regions, rural areas, and patients who did not live alone had more awareness of health management services (OR= 0.347, 3.563, and 0.281, all P <0.05). The overall utilization rate of health management services for chronic diseases in chronic disease patients was 94.58%. Multivariate Logistic regression analysis revealed that patients who lived in developed regions and rural areas showed better utilization of health management services (OR= 0.297 and 4.145, all P<0.05). The average satisfaction score of patients for all items of health management services was above nine. The satisfaction scores of chronic disease patients in rural areas regarding service convenience, service environment, waiting time, and service attitude were better than those of chronic disease patients in urban areas (t=-2.157, -3.614, -3.164, -2.879, all P<0.05). The overall satisfaction score regarding health management services for chronic diseases was 92.95%. Multivariate Logistic regression analysis showed that patients who purchased health insurance for urban and rural residents reported higher overall satisfaction with health management services for chronic diseases compared to those patients who purchase other medical insurance (OR=2.943, P<0.05). Conclusion The current level of awareness, utilization, and satisfaction with health management services for chronic diseases among chronic disease patients in Zhejiang province should be further improved. Moreover, attention should be paid to the balanced development of urban and rural areas to further increase the equality level of basic public health services.

Objective To analyze the spatial-temporal characteristic of the incidence of hepatitis E (HEV) in Zhejiang Province.Methods Data related to HEV cases in Zhejiang Province in 2015 were collected from the Infectious Disease Reporting Information System in the Information System of Disease Prevention and Control of China.The ArcGIS (10.0) was used to analyze the spatial autocorrelation,R software was used to explore the spatialtemporal-multicomponent of the incidence based on the epidemic component,autoregressive component and endemic component.Results There were 1 738 cases identified and the incidence of HEV in Zhejiang Province in 2015 was 3.16/105.There were obvious regional clustering of the cases (P ＜ 0.05),the northwest regional of Zhejiang had higher incidence of aggregation.The impact of the epidemic component was strong in most of the districts of Hangzhou Municipality,parts of Huzhou and Jiaxing Municipality.The impact of autoregressive component was strong in the main urban area of Hangzhou Municipality,Lin'an and Wenlin.The impact of endemic component was relatively lower and autoregressive component was the same in a whole area.Conclusion The spatial-temporal characteristic of the incidence of HEV in different areas are heterogeneous,it is suggested that the risk factors maybe different and targeted strategies should be taken to control the disease.

Objective To understand the distribution of the severe fever with thrombocytopenia syndrome (SFTS) in Zhejiang province,and predict the incidence and the probability of SFTS outbreak.Methods Based on the cases of SFTS from 2011-2015,software ArcGIS 10.0 was used to analyze the spatial distribution,Moran's I and Getis-Ord Gi were used to analyze the spatial autocorrelation.The incidence trend was explored by trend surface analysis,and the prediction was made by Kriging interpolation.Results The incidence of SFTS increased and the distribution expanded in Zhejiang from 2011 to 2015,the seasonal and the demographic characteristics of SFTS were similar to the previous research;there were regional clustering of the cases (P＜0.001);a downward trend was observed from northeast to southwest in terms of incidence of SFTS;the second-order disjunctive Kriging interpolation based on circular model and the indicator Kriging interpolation based on exponential model had higher prediction accuracy,the probabilities of outbreak in Anji,Daishan and Tiantai were high,the prediction deviation of inland was less than that of edge area.Conclusion The prediction of SFTS by Kriging interpolation had high accuracy,the incidence of SFTS was higher and the distribution of SFTS was larger than the results of surveillance,the risk areas for epidemic were Anji,Daishan,Ninghai,Tiantai,Sanmen and Linhai.

Objective:To investigate the inhibitory effect of combined strategy of poly adenosine diphosphate ribose polymerase (PARP) inhibitor and interleukin-1β (IL-1β) inhibitor on homologous recombination deficiency (HRD)-proficient ovarian cancer cells.Methods:(1) HRD-proficient ovarian cancer cell lines OVCAR3 and CAOV3 were treated with PARP inhibitor olaparib. Screening by RNA sequencing analysis, the expression level of IL-1β was validated by enzyme-linked immunosorbent assay (ELISA) and western blot. (2) The dose-response curves of IL-1β inhibitor diacerein were evaluated by cell counting kit-8 (CCK-8) assays in OVCAR3 and CAOV3 cells. CCK-8 assays were further applied to determine the viabilities of OVCAR3 and CAOV3 cells. (3) To evaluate the synergistic effects of olaparib and IL-1β inhibitor in vivo, the transplanted ovarian cancer model was constructed. BALB/c-nude mice ( n=16) were injected intraperitoneally with 1×10 7 OVACR3 cells labelled with luciferase (OVCAR3-Luc). Immunohistochemistry (IHC) assay was performed to determine nuclear antigen associated with cell proliferation (Ki-67) expression. (4) Blood routine tests, kidney and liver function tests were performed to analyze the toxic reaction of different drug treatments. The potential drug-induced injuries of vital organs including heart, liver, spleen, lungs and kidneys of nude mice were determined by hematoxylin-eosin (HE) staining. Results:(1) The RNA sequencing results showed that the mRNA level of IL-1β was the most significantly increased among the 25 differentially expressed genes in OVCAR3 cells treated with olaparib, compared to the negative control group. Olaparib treatment significantly promoted the secretion and expression of IL-1β protein in both OVACR3 and CAOV3 cells by ELISA [(36.2±3.5) and (49.5±3.5) pg/ml, respectively; all P<0.001] and western bolt (2.87±0.37 and 2.05±0.08, respectively; all P<0.01). (2) The half maximal inhibitory concentration (IC 50) value of IL-1β inhibitor was determined as follows: 75 μmol/L for OVACR3 cells and 100 μmol/L for CAOV3 cells. The treatments were divided into four groups including control group, olaparib monotherapy group, IL-1β inhibitor monotherapy group and the combination therapy group. The cell viabilities of each group in OVCAR3 and CAOV3 were determined by CCK-8 assay. The data in each group were showed as follows for OVCAR3 and CAOV3 cells: (100.0±0.4)% and (100.0±3.5)% in control group; (63.1±6.2)% and (63.3±3.8)% in olaparib monotherapy group; (61.6±4.7)% and (63.8±3.5)% in IL-1β inhibitor monotherapy group; and (32.9±5.2)% and (30.0±1.3)% in the combination therapy group. The viability assay showed that the combined strategy exhibited a significant inhibition effect on OVACR3 and CAOV3 cells, compared to the monotherapy group and the control group (all P<0.01). (3) All mice with transplanted tumors of HRD-proficient ovarian cancer cells were randomly divided into four groups, and treated with four different treatments as mentioned above, respectively. After 4 weeks (on day 29), the vivo fluorescence imaging were determined. The results showed that the amount of fluorescence of transplanted tumors was mostly decreased in the combination therapy group [(0.5±0.4)×10 10 p/s], compared to the control group [(4.2±1.0)×10 10 p/s] or the groups treated with any single drug [(3.1±0.9)×10 10, (2.2±0.9)×10 10 p/s; all P<0.05]. Mice were then sacrificed under anesthesia, and all transplanted tumors detached and weighed for further investigation. The weight of transplanted tumors was significantly decreased in the combination therapy group [(0.09±0.03) g], compared to that in control group [(0.25±0.05) g] or groups treated with any single drug [(0.17±0.03), (0.19±0.04) g; all P<0.05]. The measurement of the expression of Ki-67 showed that it was significantly decreased in the combination therapy group (0.33±0.10), compared to that in the control group (1.00±0.20) or monotherapy groups (0.76±0.07, 0.77±0.12; all P<0.05). (4) There were no significant differences of body weights, blood routine test, renal and liver function tests among mice with different treatments (all P>0.05). Moreover, no significant injuries were observed in the vital organs among the four groups. Conclusions:The combination of olaparib and IL-1β inhibitor synergistically exhibits significant cytotoxicity in HRD-proficient ovarian cancer cells. Moreover, the blood routine and blood biochemistry results confirmed the biosafety of the combination of olaparib and IL-1β inhibitor.

OBJECTIVE: To explore the genetic basis for a child with Rothmund-Thomson syndrome (RTS). METHODS: The child has featured poikeloderma, short stature, cataract, sparse hair and skeletal malformation. Peripheral blood samples of the child and her family members were collected and subjected to whole exome sequencing. Candidate variants were verified by Sanger sequencing. RESULTS: The child was found to harbor compound heterozygous variants of the RECQL4 gene, namely c.1048_1049delAG and c.2886-1G>A, among which c.2886-1G>A was unreported previously. According to the ACMG guidelines, the c.1048_1049delAG was predicted to be pathogenic (PVS1+PM3_Strong+PM2), while the c.2886-1G>A was predicted to be likely pathogenic (PVS1+PM2). CONCLUSION The compound heterozygous variants of the RECQL4 gene probably underlay the pathogenesis of RTS in this patient. Above finding has enriched the mutational spectrum of the RECQL4 gene.
Child ; Family ; Female ; Humans ; Mutation ; RecQ Helicases/genetics* ; Rothmund-Thomson Syndrome/genetics* ; Whole Exome Sequencing