1.A rapid GeXP-based multiplex reverse transcription-PCR assay for simultaneous detection of 5 subtypes of diarrheogenic Escherichia coli
Jiancai CHEN ; Honghu CHEN ; Yunyi ZHANG ; Junyan ZHANG ; Zheng ZHANG ; Junhang PAN ; Li ZHAN
Journal of Preventive Medicine 2022;34(10):1075-1080
Objective:
To establish a rapid GeXP-based multiplex reverse transcription-PCR assay (GeXP assay) for simultaneous detection of 5 subtypes of diarrheogenic Escherichia coli.
Methods:
Specific primers were designed according to reserved sequences of 12 virulence genes in enterotoxigenic E. coli (ETEC), enterinvasive E. coli (EIEC), enteropathogenic E. coli (EPEC), enteroaggregative E. coli (EAEC) and enterohemorrhagic E. coli (ETEC), and PCR amplification was performed with a single pair of primers to validate the specificity of PCR assay with a single template and a single pair of primers. The specificity of the GeXP assay was evaluated with the genomic DNA of 5 subtypes of diarrheogenic E. coli as the template in a mixture of 12 pairs of primers, and the sensitivity of the GeXP assay was evaluated with the mixed suspensions of 5 subtypes of diarrheogenic E. coli at concentrations of 106, 105, 104 and 103 CFU/mL as the template. Foods purchased from supermarkets and agricultural retail markets were prepared into 34 spiked samples, and 5 subtypes of diarrheogenic E. coli were detected using the GeXP assay and compared with the fluorescent real-time multiple PCR assay.
Results:
The sizes of sth, pic, bfpB, astA, lt, escV, aggR, stx1, uidA, invE, stx2 and stp genes amplification products were consistent with expected sizes using a single template and a single pair of primers, with a fluorescent signal intensity of more than 25 000 A.U. The sizes of the GeXP assay amplification products of 12 virulence genes in 5 subtypes of diarrheogenic E. coli were consistent with expected sizes, with a high specificity. If the concentration of the mixed suspensions of 5 subtypes of diarrheogenic E. coli was 103 CFU/mL, the GeXP assay was effective for simultaneous detection of 12 virulence genes, with a high fluorescent signal intensity, consistent repeated detection results and a less than 10% coefficient of variation. The GeXP assay detected 3 ETEC isolates, 12 EAEC isolates, one EIEC isolate, one EPEC isolate and one EHEC isolate among the 34 spiked samples, which was in agreement with the detection of 5 subtypes of diarrheogenic E. coli with commercial fluorescent real-time multiple PCR assay kits.
Conclusions
A GeXP assay has been successfully established for simultaneous detection of 12 virulence genes in diarrheogenic E. coli, which is effective for clinical differential diagnosis and epidemiological surveys of diarrheogenic E. coli.
2.The semi-quantitative risk assessment of Listeria monocytogenes contamination in cooked meat products in bulk in Zhejiang Province
ZHANG Junyan ; ZHAN Li ; ZHANG Yunyi ; CHEN Jiancai ; CHEN Honghu ; ZHANG Zheng
Journal of Preventive Medicine 2021;33(7):656-660
Objective:
To investigate the contamination status and assess the potential consumption risk of Listeria monocytogenes ( L. monocytogenes ) in cooked meat products in bulk in Zhejiang Province, so as to provide strategy for food safety supervision and management.
Methods:
A total of 2 320 cooked meat products were sampled from eleven cities in Zhejiang Province during 2018-2020. The detection of L. monocytogenes was carried out in accordance with the national standard GB/T 4789.30-2016. Risk Ranger software was used for the semi-quantitative risk assessment on the whole population and pregnant women.
Results:
The total detection rate of L. monocytogenes in cooked meat products in bulk in Zhejiang Province was 2.97% ( 69/2 320 ). The detection rates in stewed, smoked/roasted, fried, dried products and others were 3.85%, 1.81%, 0.59%, 0% and 0.94%, which were significantly different ( P<0.05 ). There were 28 positive samples in 1 069 samples collected in 2020, with the concentration ranging from 5 to 590 CFU/g and averaging 6.8 CFU/g. The estimated number of listeriosis cases each year caused by consumption of cooked meat products in bulk was 131 in the whole population with a risk score of 42, and 1.44 in pregnant women with a risk score of 54. The risk coefficient could reduce to approximate zero after sufficient heating before intake.
Conclusion
The prevalence of L. monocytogenes in cooked meat products in bulk in Zhejiang Province during 2018-2020 poses a potential risk in food safety. Pregnant women should avoid eating.
3.Analysis of the status of professional self-efficacy and its influencing factors among postgraduates in Master of Nursing Specialist
Qiaoqiao SHEN ; Pei TONG ; Yunyi LIANG ; Jingjing ZHOU ; Yajing ZHENG ; Yulin GAO
Chinese Journal of Practical Nursing 2021;37(11):825-830
Objective:To investigate the status of professional self-efficacy and its influencing factors among postgraduates in Master of Nursing Specialist(MNS).Methods:Totally 185 postgraduates from 33 universities were chosen by convenience sampling from December 2018 to January 2019 for a cross-sectional survey. Subjects completed a questionnaire covering demographic data, Professional Self-efficacy Questionnaire for Nursing Students, the Chinese version of Proactive Personality Scale and Career Social Support Inventory for Chinese College Students.Results:The score of professional self-efficacy was (92.78±13.22) . Multiple linear regression analysis showed that the sense of achievement in clinical work, proactive personality and career decision support were the influencing factors of postgraduates professional self-efficacy, with an interpretability of 40.2%.Conclusion:Professional self-efficacy of postgraduates is at upper middle level. Schools, hospitals and other relevant institutions should further improve the postgraduates cultivation model, establish high quality clinical practice bases, encourage postgraduates to actively explore career development pathway and provide adequate advices and supports, to improve the professional self-efficacy of postgraduates.
4.Effect of Matteuccia struthiopteris polysaccharides on systemic lupus erythematosus-like syndrome induced by Campylobacter jejuni in BALB/c mice
Zheng WANG ; Junyun XIE ; Han XU ; Xiaoqin CHENG ; Xiling YUE ; Hong LI ; Yunyi ZHANG ; Yan LU ; Daofeng CHEN
Acta Pharmaceutica Sinica 2010;45(6):711-7
Matteuccia struthiopteris is a nature plant, which contains a lot of potential active components. In the present study, we investigated the effect of polysaccharides extracted from Matteuccia struthiopteris on lupus-like syndrome induced by Campylobacter jejuni CJ-S131 in BALB/c mice. Mice were randomly divided into normal, model control, SLE model (vehicle treated), Matteuccia struthiopteris polysaccharides treated (30 and 15 mg x kg(-1)) groups and prednisone 5 mg x kg(-1) treated groups. The effect of Matteuccia struthiopteris polysaccharides (Ms) on weight and organ index of BALB/c mice was detected. Autoantibodies and total IgG production were measured by enzyme linked immunosorbent assay. Proteinuria was measured and kidneys were examined by light microscopy. Compared with SLE model group, treatment with Matteuccia struthiopteris polysaccharides 30 and 15 mg x kg(-1) reduced weight loss and Matteuccia struthiopteris polysaccharides 15 mg x kg(-1) reduced spleen swelling (P < 0.05). The increased production of autoantibodies and total immunoglobulin G (IgG) were also significantly inhibited. Matteuccia struthiopteris polysaccharides protected kidney against glomerular injury in BALB/c mice with reduced immunoglobulin deposition and lowered proteinuria (P < 0.01). Matteuccia struthiopteris polysaccharides had a protective effect on lupus-like syndrome induced by CJ-S131 in BALB/c mice.
5.Alterated Serum Levels of Immunoglobulin E in Patients With Calcific Aortic Valve Disease
Yunyi ZHENG ; Qingchun ZENG ; Qiong ZHAN ; Dingji ZHU ; Jinghai HUA ; Haobin ZHOU ; Dongqi AN ; Zuheng LIU ; Wenyan LAI ; Dingli XU
Chinese Circulation Journal 2016;31(3):263-266
Objective: To explore the relationship between serum levels of immunoglobulin E (IgE) and calciifc aortic valve disease (CAVD) in relevant patients.
Methods: A total of 394 patients were enrolled in our study. Based on echocardiography presentation, the patients were divided into 2 groups: CAVD group,n=169 and Non-CAVD group,n=225. Serum levels of IgE were examined by chemiluminescence method. The IgE levels were compared between 2 groups and the relationship between serum IgE level and CAVD was analyzed.
Results: Serum levels of IgE in CAVD group was significantly higher than Non-CAVD group 113.30 IU/ml vs 63.76 IU/ml (P<0.05); multivariate logistic regression analysis conifrmed above difference (P<0.05) and it also indicated that the alteration of surum IgE level is obviously related to CAVD occurrence.
Conclusion: Serum IgE level is obviously increased in CAVD patients. IgE is an independent biochemical indicator of CAVD, it may play the important role in CAVD pathogenesis.
6.Real-time recombinase polymerase amplification for detection of Vibrio parahaemolyticus
Li ZHAN ; Changping XU ; Yunyi ZHANG ; Honghu CHEN ; Zheng ZHANG ; Jiancai CHEN ; Junyan ZHANG ; Lingling MEI
Journal of Preventive Medicine 2019;31(7):653-657
Objective:
To establish real-time recombinase polymerase amplification(RPA)for the rapid detection of Vibrio parahaemolyticus(VP).
Methods:
An exo probe and primers were designed according to the conserved sequence of thermolabile hemolysin(tlh)gene of VP and then RPA for detection of VP was established. The sensitivity of the assay was evaluated by detecting different concentration of VP;the specificity was evaluated by detecting different bacteria;the stability was evaluated by repeat trials;the application effect was evaluated by detecting food samples which were simultaneously tested with traditional culture method according to GB 4798.7-2013 Detection of VP.
Results:
A real-time RPA was established to complete VP amplification within 20 min at a constant temperature of 39 ℃. The analytical sensitivity of the assay was five pg per reaction and no cross-reactivity with other pathogenic bacteria observed. The RPA detection results with different concentration of VP and E. coli DNA templates at three time points were consistent. The detection results of 51 food samples by RPA were the same as those by traditional culture method.
Conclusion
The established real-time RPA can qualitatively detect VP,with simple operation and interpretation of results,which is suitable for rapid detection of VP in public health emergencies and food safety supervision.
7. Cross-cultural adjustment of successful aging scale and its reliability and validity in urban elderly
Pei TONG ; Jingjing ZHOU ; Yajing ZHENG ; Qingzhuo REN ; Yunyi LIANG ; Qiaoqiao SHEN ; Xiaojia SUN ; Yulin GAO
Chinese Journal of Behavioral Medicine and Brain Science 2019;28(11):1030-1036
Objective:
To revised the Taiwan version successful aging inventory (SAI), and verify its reliability and validity in urban elderly in mainland China.
Methods:
SAI was adjusted by culture and revised by pre-test to form a formal questionnaire. A sample of 600 elderly people in Guangzhou was chosen for a face-to-face questionnaire survey through convenient sampling. Half of the sample (
8.Evaluation of precision rehabilitation services for people with disabilities in China
Chinese Journal of Rehabilitation Theory and Practice 2022;28(10):1117-1125
ObjectiveTo evaluate the implementation of the precision rehabilitation services for people with disabilities in China. MethodsData from the Second China National Sample Survey on Disability, 2017-2021 China Statistical Yearbook on the Work for Persons with Disabilities and other official statistics were used to evaluate the precision rehabilitation services for people with disabilities in China from four aspects: the construction of organizational system, the arrangement of rehabilitation management system, the progress of rehabilitation service capacity and the utilization of rehabilitation services. ResultsA top-down precision rehabilitation organization system was basically established; a rehabilitation management system was used throughout the country, while the operation specification and data quality needed to be optimized; the capacity of rehabilitation services was steadily improved to meet the diverse rehabilitation needs of people with disabilities; and more and more disabled children and disabled persons with certificates received basic rehabilitation services, and the accuracy of rehabilitation services was constantly improved. ConclusionThe precision rehabilitation services for people with disabilities in China is working well in the early stage. It is needed to expand resources and improve the ability of rehabilitation services, to strengthen the economic guarantee and summarize typical cases, to improve public awareness of rehabilitation. The government should improve the sense of responsibility and working mechanism, to promote the coordinated development of rehabilitation services among regions, and develop community-based rehabilitation vigorously.
9.Serum miR-149-5p and matrix metalloproteinase-9 predict hemorrhagic transformation after intravenous thrombolysis in patients with acute ischemic stroke
Xianjun HE ; Ming ZHAO ; Yu KONG ; Yunyi DAI ; Xiuxia ZHENG
International Journal of Cerebrovascular Diseases 2022;30(9):652-656
Objective:To investigate the correlation and predictive value of serum miR-149-5p and matrix metalloproteinase-9 (MMP-9) and hemorrhagic transformation (HT) after intravenous thrombolysis in patients with acute ischemic stroke (AIS).Methods:Patients with AIS received intravenous thrombolytic therapy in Shangqiu First People's Hospital from September 2019 to February 2022 were enrolled prospectively. They were divided into HT group and non-HT group according to whether HT occurred after intravenous thrombolysis. Serum miR-149-5p and MMP-9 were measured by real-time fluorescence quantitative polymerase chain reaction and enzyme-linked immunosorbent assay respectively. Multivariate logistic regression analysis was used to determine the independent risk factors for HT after thrombolysis. The receiver operating characteristic (ROC) curve was used to evaluate the predictive value of serum miR-149-5p, MMP-9 and their combination for HT after intravenous thrombolysis. Results:A total of 358 patients with AIS received intravenous thrombolytic therapy were enrolled, 71 of them (19.83%) developed HT. The serum MMP-9 in the HT group was significantly higher than that in the non-HT group (273.95±35.23 μg/L vs. 202.71±30.52 μg/L; t=17.062, P<0.001), while the serum miR-149-5p was significantly lower than that in the non-HT group (0.26±0.06 vs. 1.03±0.15; t=42.387, P<0.001). Multivariate logistic regression analysis showed that atrial fibrillation (odds ratio [ OR] 2.282, 95% confidence interval [ CI] 1.731-3.008; P<0.001), time from onset to intravenous thrombolysis ( OR 2.334, 95% CI 1.458-3.735; P<0.001), miR-149-5p ( OR 1.758, 95% CI 1.142-2.705; P=0.010) and MMP-9 ( OR 1.535, 95% CI 1.106-2.129; P=0.010) were the independent risk factors for HT after intravenous thrombolysis. Serum miR-149-5p (area under the curve 0.856, 95% CI 0.803-0.909; when the optimal cut-off value was 0.741, the sensitivity was 80.3% and the specificity was 89.9%), MMP-9 (area under the curve 0.875, 95% CI 0.821-0.929; when the optimal cut-off value was 240.051 μg/L, the sensitivity was 83.1% and the specificity was 90.2%) and their combination (area under the curve 0.897, 95% CI 0.854-0.941; sensitivity 84.5% and specificity 90.6%) had better predictive value for HT after thrombolysis, and there were no significant differences in the predictive value among the three. Conclusions:After intravenous thrombolysis, the serum miR-149-5p is lower and MMP-9 is higher at admission in patients with HT in patients with AIS. Both of them and their combination have better predictive value for HT after intravenous thrombolysis.
10.Risk factors and survival analysis of acute rejection in recipients after simultaneous pancreas-kidney transplantation
Rongxin CHEN ; Luhao LIU ; Jiali FANG ; Guanghui LI ; Lu XU ; Peng ZHANG ; Yunyi XIONG ; Jialing WU ; Junjie MA ; Zheng CHEN
Chinese Journal of Organ Transplantation 2023;44(12):735-742
Objective:To summarize the clinical characteristics and risk factors of acute rejection(AR)of transplanted pancreas and kidney after simultaneous pancreas-kidney transplantation(SPK)and explore the effects of AR on the survival of transplanted pancreas, kidney and recipients.Methods:From September 2016 to July 2022, the relevant clinical data were retrospectively reviewed for 218 recipients undergoing SPK.According to whether or not AR occurred after SPK, they were assigned into two groups of AR(n=53)and non-AR(n=165). The relevant clinical data were compared for two groups of donors and recipients and the risk factors of AR analyzed by binary Logistic regression.Kaplan-Meier method was employed for comparing the survival rates of recipients/transplanted pancreas and kidneys in two groups.Results:A total of 53 cases(24.3%)developed ARs of transplanted pancreas(n=31, 14.2%)(5 of 2 ARs), transplanted kidney(n=15, 6.9%)(1 of 2 ARs)and transplanted pancreas & kidney AR(n=11, 5.0%)(2 of 2 ARs). Tacrolimus blood levels in AR and non-AR groups were(5.8±1.2)and(6.3±1.6)μg/L and failed to attain targets in 36(67.9%)and 78(47.3%)cases.During follow-ups, the incidence of pneumonia and urinary tract infections in AR group versus non-AR group were[43.4%(23/53)vs.27.3%(45/165)and 39.6%(21/53)vs.18.8%(31/165)]and the differences were statistically significant( P=0.028 & 0.002). The results of multifactorial regression analysis revealed that sub-optimal blood level of tacrolimus was an independent risk factor for an occurrence of AR in grafts of SPK recipients( OR=2.254, 95% CI: 1.167-4.353, P=0.016). Comparisons of 1/5-year postoperative survival rates between recipients in AR and no-AR group(98.1% vs.93.9% and 92.1% vs.92.4%)indicated that the differences were not statistically significant( P=0.233 & 0.806). Through comparing 1/5-year survival rates of transplanted pancreas in AR and non-AR groups(94.3% vs.100%, 89.4% vs.98.6%), the differences were statistically significant( P=0.003 & 0.004). And 1/5-year survival rates of transplanted kidneys in AR and non-AR groups(92.5% vs.100% and 90.2% vs.100%)were compared and the differences were statistically significant(all P<0.001). Conclusions:The incidence of AR is higher in transplanted pancreas and kidney after SPK.And the incidence of pneumonia and urinary tract infection is higher in AR group than that in non-AR group.Sub-optimal blood level of tacrolimus is an independent risk factor for the occurrence of AR.The 1/5-year survival rates of transplanted pancreas and transplanted kidney are lower in AR group than those in non-AR group.It has some effect on the survival of transplanted pancreas and kidney.