OBJECTIVE To investigate the existance of class Ⅰ gene integron and transposon(merA and tnpA) gene marker in Escherichia coli isolated from the elderly patients.METHODS Antibiotic sensitivity was performed by microdilution test.Integron gene IntⅠ1 and transposon marker genes were detected by polymerase chain reaction(PCR).The PCR products were purified and sequenced.RESULTS E.coli resistant to amikacin accounted for 60%,to sulfamethoxazole 95%,and to third-generation cephalosporin 100%(all ESBLs-producing strains).The resistance was related closely to the class Ⅰ integron.The positive rate of integron intⅠ1 was 80%(16/20),whereas the positive rate of gene merA was only 10%(2/20).Gene tnpA was not tested out.CONCLUSIONS The drug resistance of E.coli has close relation with transmission of integron and transposon.The transposons of Tn21/Tn501 are all firstly reported in China.

OBJECTIVE To investigate the ?-lactamases in Klebsiella pneumoniae strains isolated from the elderly patients.METHODS We had an observation on 20 strains of Klebsiella pneumoniae about 15 kinds of ?-lactamase gene test(blaTEM,blaSHV,blaLEN,laOKP,blaCTX-M-1 group,blaCTX-M-2 group,blaCTX-M-9 group,blaOXA-1 group,blaOXA-2 group,blaOXA-10 group,blaGES,blaPER,blaVEB,blaDHA,and blaACT-1).RESULTS Six kinds of ?-lactamase genes had been found out from these 20 strains of K.pneumoniae,namely,blaTEM,blaSHV,blaCTX-M-1 group,blaCTX-M-9 group,blaOXA-1 group,and blaDHA.Their positives were 95%,30%,50%,5%,5% and 15%.Among these 20 strains of K.pneumoniae,there were 19 ones of K.pneumoniae which at least contained 1 kind of ?-lactamase genes.Fifteen strains of K.pneumoniae were found containing more than 2 kinds of ?-lactamase genes,at the same time.Some were even tested out having 4 kinds of ?-lactamase genes.CONCLUSIONS The resistance of ?-lactamase antibiotics of these 20 strains of K.pneumoniae has a close relative to the number of ?-lactamase.

OBJECTIVE To investigate the chloramphenicol-resistant gene in Escherichia coli strains isolated from the old patients.METHODS Twenty strains of E.coli were clinically isolated.The resistance genes were analyzed by polymerase chain reaction(PCR) and verified by DNA sequencing.RESULTS The positive rates of catB and cmlA were 20% and 20%,respectively.Two strains of E.coli were found containing catB gene and cmlA gene.CONCLUSIONS The resistance to chloramphenicol in these 20 strains of E.coli has a close relation to antimicrobial-resistance gene existed,and the genes of catB and cmlA in E.coli are all firstly found and reported in China.

OBJECTIVE To investigate the prevalence of sulfamethoxazole/trimethoprim(SXT)-resistance genes and drug resistance in Escherichia coli isolates from the old patients.METHODS Drug resistance of 20 E.coli strains were analyzed.The SXT-resistance genes(sul1,dfrA1,dfrA12 and dfrA17)were detected by polymerase chain reaction(PCR).The PCR products were purified and sequenced.RESULTS In 20 E.coli strains,95.0% were SXT resistant.The positive rate of genes of sul1,dfrA12 and dfrA17 were 80.0%,20.0% and 65.0%.Gene of dfrA1 was not detected out,17 strains of E.coli were found dfrA gene.The SXT-resistance gene dfrA or sul1 was detected in 18 of the 20 strains.CONCLUSIONS The SXT resistance of E.coli can be caused mainly by gene expression of dfrA and sul1,and the genes of sul1,dfrA12 and dfrA17 are all firstly found and reported in China.

Objective: To investigate the clustering of risk factors among populations at high risk of cardiovascular diseases in Shaoxing City, Zhejiang Province, so as to provide the evidence for formulating the cardiovascular disease control measures among populations at high risk of cardiovascular diseases. Methods: The populations with hypertension, diabetes, dyslipidemia and a history of cardio-cerebrovascular disease at ages of 35 to 70 years were sampled from 6 districts (counties) of Shaoxing City using a multi-stage cluster random sampling method from May to July 2021. Participants' demographics, history of disease, smoking, alcohol consumption and drug use were collected through questionnaires, and height, body weight and blood lipid were measured. The participants at high risk of cardiovascular diseases were enrolled, and the prevalence and clustering of five risk factors were investigated, including current smoking, current alcohol consumption, overweight/obesity, never exercise and daily sedentary duration of 3 hours and longer. Factors affecting the clustering of risk factors were identified with an ordinal logistic regression model. Results: Totally 1 695 participants were enrolled, including 940 men (55.46%) and 755 women (44.54%), with a mean age of (62.56±6.08) years old. There were 213 participants with a history of cardio-cerebrovascular disease (12.57%), and the prevalence of hypertension, diabetes and dyslipidemia was 78.53%, 61.95% and 46.78%, respectively, and there were 32.63% of participants with current smoking, 35.99% with current alcohol consumption, 49.38% with overweight/obesity, 62.12% with never exercise and 61.24% with daily sedentary duration of 3 hours and longer, respectively. There were 28.85% of participants with two risk factors, and 46.90% with three to five risk factors. Ordinal logistic regression analysis showed that male (OR=5.430, 95%CI: 4.389-6.726), and development of hypertension (OR=1.655, 95%CI: 1.313-2.090) led to present more numbers of clustering of risk factors, and annual household income of 20 000 to 50 000 Yuan (OR=0.620, 95%CI: 0.473-0.812), a history of cardio-cerebrovascular diseases (OR=0.430, 95%CI: 0.324-0.572), presence of diabetes (OR=0.592, 95%CI: 0.476-0.736) led to less numbers of clustering of risk factors among populations at high risk of cardiovascular diseases. Conclusions There is clustering of risk factors among populations at high risk of cardiovascular diseases in Shaoxing City. Men, income, history of cardio-cerebrovascular diseases, hypertension and diabetes are factors affecting the clustering of risk factors.

Objective: To investigate sleep quality among patients with hypertension and diabetes, so as to provide the basis for improving the health of patients with chronic diseases. Methods: From May to August 2022, patients with hypertension and diabetes under the community management in 4 counties and cities (districts) of Shaoxing City, Zhejiang Province were recruited using a multi-stage stratified cluster random sampling method. Demographic information, disease history, smoking, alcohol consumption, physical activity and sleep quality were collected by questionnaire surveys, and blood pressure, fasting blood glucose and glycosylated hemoglobin were collected through physical examination and laboratory testing. The incidence of poor sleep quality symptoms (snoring/asphyxia/suffocation, difficulty falling asleep, night awakening, taking sleeping pills and early morning awakening) in patients with diabetes and hypertension was descriptively analyzed, and sleep quality was evaluated. Results: A total of 1 539 patients with hypertension and diabetes were surveyed, including 715 males (46.46%) and 824 females (53.54%). The patients had a mean age of (68.68±7.62) years. The proportions of night awakening, early morning awakening, snoring/asphyxia/suffocation, difficulty falling asleep and taking sleeping pills were 29.82%, 28.53%, 20.73%, 15.79% and 4.29%, respectively. The proportion of poor sleep quality was 66.54%. Employment status, smoking status, moderate/high intensity physical activity and blood pressure control were related to poor sleep quality (all P<0.05). Conclusion Poor sleep quality is common in patients with hypertension and diabetes, with the main symptoms being night awakening, early morning awakening, difficulty falling asleep and snoring/asphyxia/suffocation.

Aim To investigate the effect of genistein on apoptosis in triple-negative breast cancer MDA-MB-231 cells and the underlying mechanisms.Methods MTT assay was used to detect the inhibition rate on breast cancer MDA-MB-231 cells of genistein.Hoechst 33258 staining was applied to determine the effect of genistein on morphology of MDA-MB-231 cells.qRT-PCR was employed to detect the mRNA expression of EGFR in MDA-MB-231 cells.Western blot was utilized to determine the expression of Bcl-2, Bax, caspase-3, EGFR, Akt, and p-Akt.The expressions of Akt and p-Akt proteins in breast cancer MDA-MB-231 cells were detected after treated with Akt activator insulin, genistein and in combination with insulin.Results Genistein inhibited the viability of breast cancer MDA-MB-231 cells in a time-dependent manner.The results of Hoechst 33258 staining showed a typical apoptotic morphological changes of MDA-MB-231 cells after treatment of genistein for 36 h.qRT-PCR showed that the mRNA expression of EGFR in MDA-MB-231 cells decreased after treated with genistein for 36 h.The expression levels of Bcl-2, EGFR, Akt, p-Akt, ERK, p-ERK were significantly down-regulated(P<0.01) compared with control.While, the expression of Bax, caspase-3 was significantly up-regulated (P<0.01).It was observed that p-Akt was significantly activated after the treatment of Akt activator insulin (P<0.01), however, significantly down-regulated (P<0.01) when treated with genistein.Conclusion Genistein could inhibit the growth of triple-negative breast cancer MDA-MB-231 cells and induce apoptosis, which probably involves regulating EGFR/PI3K/Akt signaling pathway.

Objective:To analyze the characteristics of the molecular transmission network of newly-diagnosed human immunodeficiency virus type 1 (HIV-1) infected individuals in Shaoxing City, Zhejiang Province, and to provide evidence for epidemic trend and prevention.Methods:The plasma samples from 423 antiretroviral-naive HIV-1/acquired immunodeficiency syndrome patients from August 2018 to December 2019 were collected, and the pol gene fragments of HIV-1 from 375 samples were amplified by reverse transcription polymerase chain reaction (PCR) and nested PCR. The phylogenetic tree was constructed to analyze the molecular transmission network for subtypes and different gene distances by MEGA 6.0 software, HyPhy software and Cytoscape 3.7.2. Mutations on drug resistance was analyzed by online software tool of the HIV drug resistance database of Stanford University. Results:Eight subtypes were found in the 375 samples. Circulating recombinant form (CRF)07_BC(215/375, 57.33%) and CRF01_AE(103/375, 27.47%) were the major subtypes, followed by CRF85_BC, CRF55_01B, B, C, and CRF01_AE/B subtypes. One hundred and ninety-four individuals (51.73%) were connected to the transmission network at 1.50% genetic distance with 24 clusters. One hundred and twenty-nine individuals (34.40%) were connected to the transmission network at 0.75% genetic distance with 30 clusters, and 35 elderly patients were clustered in CL1.Forty-two cases had surveillance drug resistance mutation (SDRM), the prevalence of transmitted drug resistance was 11.20%(42/375). Thirty-eight cases had the drug mutations to non-nucleoside reverse transcriptase inhibitor, including K103 N(32/375, 8.53%), K103 S(4/375, 1.07%), Y188 L(1/375, 0.27%) and G190 A(1/375, 0.27%); four cases had the mutations to protease inhibitor, including M46 I(3/375, 0.80%) and V82 A(1/375, 0.27%). The cluster C2 sequences carried a high proportion of resistant mutations (94.29%, 33/35). Conclusions:HIV-1 subtypes in Shaoxing City are diverse and the CRF07_BC subtype spreads rapidly. The elderly patients with drug resistance genes in cluster CL1 at 0.75% gene distance need to be intervened immediately to prevent the drug resistance virus spread.

Objective To compare the relationship between the enzyme‐linked immunosorbent assay(ELISA) reagent and West‐ern blot(WB) confirmation reagent for analyzing the quality lever of human T‐cell lymphotropic virus(HTLV) detection reagent . Methods The WB confirmation reagent was used to detect anti‐HTLV antibody in 156 human serum samples of ELISA prelimina‐ry screening positive .The ELISA cut‐off value(optimal value) was selected by using the two‐graph receiver operating characteristics (TG‐ROC) analytical method .The two‐by‐two table analysis was constructed to analyze the consistency of results detected by the two methods ,moreover the McNemar test was used to evaluate the consistency of detection results .The quality level of HTLV de‐tection reagent was comprehensively evaluated .Results Among 156 serum samples of ELISA preliminary screening positive ,only 40 samples were positive by the WB confirmation ,and other 116 samples were negative .The sensitivity and specificity of ELISA de‐tection reagent obtained by TG‐ROC analysis were 97 .5% and 45 .7% respectively ,the TG‐ROC test also indicated that the detec‐tion results had significant difference between ELISA and WB(P<0 .05) .By adjusting the cut‐off value ,the sensitivity and specific‐ity of ELISA were increased to 88 .8% (parametric method) .In the comparison of the parametric method and the non‐parametric method ,the obtained areas under the curve(AUC) was 0 .923 5(parametric method) ,their results were basically consistent .Conclu‐sion Although above results indicate that the detection results of ELISA reagent are different from those of WB ,but adjusting the cut off value can increase its sensitivity and specificity ,thus increases the reliability of diagnosis result .