1.Exploration of antibiotic resistance and population genetic characteristics of Salmonella Derby in China
Xinjiao HOU ; Huiying SUN ; Luyan WANG ; Meiying YAN ; Xuewen LI
Chinese Journal of Epidemiology 2024;45(5):730-737
Objective:To characterize the antimicrobial resistance, resistance machanism and population genetics of Salmonella( S.) Derby in China, preliminarily reveal the population genetic characteristics of S. Derby in China, discover possible transmission patterns or potential transmission pathways, and provide certain reference for strengthening S. disease monitoring and developing prevention and control strategies. Methods:A total of 201 strains of S. Derby from different areas in China were used for the susceptible tests to 16 antibiotics and whole-genome sequencing. Finally, combined with the genome sequences of 134 strains of S. Derby from public databases, 335 strains of S. Derby were used for resistance genotype analysis and multi-locus sequence typing (MLST), and a phylogenetic tree based on the core genome single nucleotide polymorphisms was constructed for evolutionary analysis. Results:The results showed that 201 strains of S. Derby showed resistance to 16 kinds of antibiotics at different levels. The overall resistance rate was 97.51%. The resistance rates to antibiotics varied in S. Derby from different sources (human, animal, and food), the differences were significant (all P<0.05). A total of 38 resistance genes were carried by 335 strains of S. Derby, of which, fosfomycin gene fosA7 was found in all the strains (100.00%) and aminoglycoside genes aac(6')-Iaa accounted for 99.70%. The consistency of resistance genes and phenotypes varied with antibiotics. Except aminoglycosides and chloramphenicol, the consistencies of resistance genes and phenotypes for other antibiotics were high. MLST showed that 334 strains of S. Derby belonged to ST40. Phylogenetic trees indicated the risk for cross-infection between animal and human, food and human, and the possibility of long-distance interprovincial transmission of the bacteria by animal, to which further epidemiological studies are needed. Conclusions:The drug resistance of S. Derby is serious in China and the risk for cross-transmission between human and animal or food exists. It is necessary to establish and strengthen the comprehensive surveillance and risk assessment to prevent the spread of antibiotic resistant strains or elements through animal, food and human chains.
2.Rapid serotyping of Salmonella based on matrix assisted laser desorption ionization-time of flight mass spectrometry
Luyan WANG ; Xinjiao HOU ; Huiying SUN ; Baowei DIAO ; Jie LI ; Meiying YAN
Chinese Journal of Epidemiology 2024;45(9):1266-1272
Objective:To establish a matrix assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) assay for the identification of common Salmonella serotypes and provide etiology evidence for the early precise treatment of salmonellosis. Methods:A total of 500 strains were collected from different regions and sources and five predominant Salmonella serotypes ( Salmonella Typhi , Salmonella Paratyphi A , Salmonella Typhimurium , Salmonella Enteritidis , and Salmonella Indiana) of each strain was identified by agglutination test and whole-genome sequencing. The protein complex of the strains was extracted by using optimized pretreatment method to establish the fingerprint database of peptides for each Salmonella serotype. The new serotyping assays were established by using different modules based on the mass spectra database. Additional 155 strains with specified serotypes and variant sources were used to test and evaluate the accuracy of the new typing assays. Results:Five MALDI-TOF MS databases were established, and two new serotyping assays were established via peptide fingerprint mapping/matching and machine learning of the neuronal convolutional network respectively based on the databases. The results showed that the fingerprint matching approach could quickly identify five common Salmonella serotypes in clinical practice compared with the machine learning method, the accuracy of fingerprint matching assay to identify five Salmonella serotypes reached 100.00% and the serotyping can be conducted within a short time (15-20 minutes) and had a good reproducibility, while the machine learning method could not completely identify these serotypes. Moreover the sensitivity and specificity of fingerprint matching assay were all 100.00% respectively, while they were only 82.23% and 95.81% for machine learning method. Conclusion:The established Salmonella serotyping assay based on MALDI-TOF MS in this study can easily, rapidly and accurately identify different serotypes of Salmonella.
3.Construction of a predictive model for postpartum hemorrhage in parturients undergoing vaginal delivery with a second child
Linlin CAO ; Caiyun WANG ; Baohua LI ; Shenglian NI ; Jie LU ; Luyan LIU ; Xiaoxiao WANG ; Zhichao GUO
Chinese Journal of Modern Nursing 2024;30(7):899-905
Objective:To explore the risk factors of postpartum hemorrhage in parturients undergoing vaginal delivery with a second child and establish a risk prediction model.Methods:Using the convenient sampling method, a total of 2 500 parturients undergoing vaginal delivery with a second child who underwent regular prenatal examinations at Peking University Third Hospital from July 2019 to March 2023 were selected as the research objects. According to the amount of blood loss, parturients with blood loss greater than or equal to 500 ml within 24 hours after delivery were selected as the case group ( n=278), while parturients with blood loss less than 500 ml within 24 hours after delivery in the same period were selected as the control group ( n=2 222). Univariate analysis and binomial Logistic regression were used to analyze the influencing factors of postpartum hemorrhage in parturients with vaginal delivery with a second child. Based on the selected risk factors, a nomogram prediction model was established using R software, and the consistency of the model was tested. Results:The incidence of postpartum hemorrhage in 2 500 parturients undergoing vaginal delivery with a second child was 11.12% (278/2 500). Binomial Logistic regression analysis showed that in in vitro fertilization-embryo transfer, pre-delivery body mass index, lateral perineal incision, neonatal weight, placenta previa, placenta implantation and manual extraction of placenta were the influencing factors for postpartum hemorrhage in parturients undergoing vaginal delivery with a second child ( P<0.05). According to the influencing factors, a nomogram model was established to predict the probability of postpartum hemorrhage in women who gave birth to a second child. The C- index of the prediction model was 0.706, the area under the receiver operating characteristic curve was 0.706, and the χ 2 value of the Hosmer-Lemeshow goodness-of-fit test was 7.720 ( P=0.461) . Conclusions:In vitro fertilization embryo transfer, pre-delivery body mass index, perineal lateral resection, neonatal weight, placenta previa, placental implantation and manual extraction of placenta are risk factors for postpartum hemorrhage in parturients undergoing vaginal delivery for a second child. The prediction model constructed based on risk factors has certain accuracy and clinical value for predicting postpartum hemorrhage in parturients with vaginal delivery of a second child.
4.The profusion evaluation method for irregular small opacities associated with pneumoconiosis in chest CT
Qiang WANG ; Jing BAI ; Luyan GE ; Ningning ZHANG ; Lijing ZHANG ; Fei KANG ; Huitai DONG
China Occupational Medicine 2024;51(6):650-655
Objective To explore a method for assessing the irregular small opacities profusion associated with occupational pneumoconiosis in chest computed tomography (CT). Methods A total of 20 occupational pneumoconiosis patients whose primary manifestation was irregular small opacities on chest digital radiography (DR) were collected as the research subjects using a retrospective study method. Comparative analysis was performed between chest DR and five mm coronal multi-planar reconstruction (MPR) of chest CT images to identify the causes of irregular small opacities. An evaluation method for the profusion of associated images of irregular small opacities in chest CT was established using technique for order preference by similarity to ideal solution-analytic hierarchy process (TOPSIS-AHP), and the results were compared against GBZ 70-2015 Diagnosis of Occupational Pneumoconiosis. Results The abnormal image distribution on the five mm coronal chest CT MPR images of the 20 patients was as follows: three cases of high-density small circular opacities, seven cases of low-density circular small opacities, six cases of diffuse low-density ground-glass opacities (GGO), four cases of reticular opacities, three cases of plate-like GGO, three cases of honeycomb opacities, and four cases of increasing lung texture. The CT values of abnormal images, from high to low were: honeycomb opacities > plate-like GGO > low-density circular small opacities > diffuse low-density GGO (all P<0.05). The consistency test results indicated that the evaluation method for the profusion of associated images of irregular small opacities in chest CT showed high level of agreement with the profusion determination criteria outlined in GBZ 70-2015 Diagnosis of Occupational Pneumoconiosis (Kappa=0.78). Conclusion Irregular small opacities observed on chest DR are formed by the superposition of multiple images of abnormal pulmonary fibrosis in patients with occupational pneumoconiosis. TOPSIS-AHP can be used to establish an evaluation method of the profusion of associated image of irregular small opacity in chest CT.
5.Caffeic acid phenethyl ester improves peritoneal dialysis-associated peritoneal fibrosis by alleviating oxidative stress injury through activating nuclear factor erythroid-2-related factor 2/heme oxygenase-1 pathway
Ying LU ; Huaying SHEN ; Luyan GAO ; Zhi WANG ; Ying ZENG ; Kai SONG
Chinese Journal of Nephrology 2023;39(6):446-455
Objective:To investigate whether caffeic acid phenethyl ester (CAPE) would improve peritoneal dialysis (PD)-associated peritoneal fibrosis by alleviating oxidative stress through activating nuclear factor erythroid-2-related factor 2 (Nrf2)/heme oxygenase-1 (HO-1) pathway.Methods:Thirty-two male Sprague-Dawley rats were randomly divided into four groups by the random number table: control (CON) group (0.9% normal saline 20 ml/d intraperitoneal injection), CAPE group (0.9% normal saline 20 ml/d+CAPE 10 mg·kg -1·d -1 intraperitoneal injection), PD group [4.25% glucose peritoneal dialysis fluid (PDF) 20 ml/d intraperitoneal injection with lipopolysaccharide 0.6 mg/kg intraperitoneal injection at day 1, 3, 5 and 7], and PD+CAPE group (CAPE 10 mg·kg -1·d -1 intraperitoneal injection in addition to PD group), with 8 rats per group. On day 28, rats were euthanized after peritoneal equilibration test, and then the parietal peritoneum and omentum were collected for follow-up tests. To further investigate the mechanism, primary peritoneal mesothelial cells (PMCs) of rats were isolated and cultured. The PMCs were stimulated with 2.5% glucose PDF and added with 5 μmol/L CAPE intervention. The Nrf2 inhibitor (ML385) was used to identify whether CAPE protected PMCs from PDF by activating the Nrf2/HO-1 pathway. Histopathological staining was used to detect structural changes of the peritoneum, and immunohistochemical analysis was performed on cleaved caspase-3, Bax, α-smooth muscle actin (α-SMA), fibronectin (FN), and typeⅠ collagen (Col-Ⅰ) protein. Western blotting was used to detect the protein expression of α-SMA, FN, transforming growth factor-β1 (TGF-β1), HO-1 and nuclear Nrf2 (N-Nrf2). The apoptosis detection kit was used to detect apoptosis and flow cytometry was used to detect reactive oxygen species (ROS) in PMCs. The malondialdehyde (MDA) and superoxide dismutase (SOD) activity detection kit were used to detect MDA content and SOD activity. Cell immunofluorescence was used to analyze the protein expression of Nrf2 in PMCs. Results:Compared with the CON group, the PD group had thicker peritoneum, and the expression levels of cleaved caspase-3, Bax, α-SMA, FN, Col-Ⅰand MDA in peritoneum were significantly higher, while HO-1, N-Nrf2 protein expression and SOD activity were lower (all P<0.05). Compared with the PD group, the parietal peritoneum morphology of CAPE+PD group was improved, accompanied by reduced cleaved caspase-3, Bax, α-SMA, FN, Col-Ⅰ protein expression, and MDA content, while N-Nrf2, HO-1 protein expression, and SOD activity were higher (all P<0.05). Compared with the CON group, the PD group had significantly lower ultrafiltration volume and higher peritoneal permeability (both P<0.05). After CAPE intervention, the peritoneal transport function of the rats was significantly improved ( P<0.05). In cultured PMCs, PDF inhibited nuclear translocation of Nrf2 and protein expression of HO-1, and upregulated intracellular ROS level. In addition, PDF increased cell apoptosis and the protein expression levels of α-SMA, TGF-β1 and FN (all P<0.05). CAPE activated nuclear translocation of Nrf2, increased HO-1 protein expression, downregulated intracellular ROS level, and partially reversed PDF-induced cell apoptosis and epithelial- mesenchymal transition (all P<0.05). The protective effects of CAPE on PMCs were partially abolished by ML385 (all P<0.05). Conclusions:CAPE can reduce PD-induced PMCs apoptosis and epithelial-mesenchymal transition by attenuating oxidative stress, and significantly improve peritoneal fibrosis and ultrafiltration function. The beneficial effects of CAPE on peritoneum are related to activation of Nrf2/HO-1 pathway.
6.Expressions and clinical significances of CD68 and ABHD5 in gastric carcinoma
Journal of International Oncology 2018;45(3):153-156
Objective To evaluate the expressions of CD68 and alpha/beta hydrolase domain-containing protein 5 (ABHD5) in gastric carcinoma and their interrelation,and the relationship with the clinicopathological features of gastric carcinoma.Methods The immunohistochemistry staining was used to determine the expressions of CD68 and ABHD5 in 60 cases of gastric carcinoma and 30 cases of adjacent normal tissues.The relationships between CD68,ABHD5 and clinicopathological features of gastric carcinoma,and the correlation of them were analyzed.Results The expressions of CD68 and ABHD5 in gastric carcinoma were significantly higher than those in adjacent normal tissues,and the positive expression rates were 65.00% vs.26.67%(x2 =11.779,P =0.001) and 60.00% vs.30.00% (x2 =7.200,P =0.007) respectively,with significant differences.The expression of CD68 was related to TNM stage (x2 =7.959,P =0.005),lymph node metastasis (x2 =6.901,P =0.009) and serosal invasion (x2 =5.833,P =0.016),but it was not associated with pathological grade (x2 =0.028,P =0.866),gender (x2 =0.533,P =0.465) and age (x2 =0.060,P =0.807).The expression of ABHD5 was related to pathological grade (x2 =4.318,P =0.038),TNM stage (/x2 =4.051,P =0.044) and serosal invasion (x2 =5.275,P =0.022),but it was not associated with lymph node metastasis (x2 =0.545,P =0.460),gender (x2 =0.191,P =0.662) and age (x2 =1.358,P =0.244).Contingency correlation analysis showed that there was positive correlation between CD68 and ABHD5 in gastric cancer (c =0.328,P =0.010).Conclusion The expressions of CD68 and ABHD5 in gastric carcinoma are higher than those in adjacent normal tissues,which are correlated with the different clinicopathological features of gastric carcinoma.The expression of CD68 is positively correlated with the expression of ABHD5.The combined detection of CD68 and ABHD5 expressions in gastric carcinoma is helpful to evaluate the invasion,metastasis and prognosis of gastric carcinoma.
7.Effects of improved TCM hot ironing technology on the gastrointestinal reaction in patients with mammary cancer
Xiaojie CHEN ; Yang YANG ; Luyan JIANG ; Yongjian WANG ; Ya′nan ZHU
Chinese Journal of Modern Nursing 2016;22(3):369-372
Objective To evaluate the effects of gastrointestinal reaction through the breast cancer chemotherapy period with improved traditional Chinese medicine ( TCM) hot ironing technology. Methods We randomly divided 80 cases of breast cancer in the chemotherapy period into control group and treatment group (40 cases in each group ). The patients of control group used Granisetron Hydrochloride Injection 3 mg, bid, for three days, while the patients of treatment group used improved TCM hot ironing technology on Zhongwan for 30 mins, qd for 3 consecutive days. The nausea, vomiting, anorexia and its degree changing were compared at chemotherapy day, 1d, 2d and 3d after chemotherapy between two groups. Results The proportion of nausea, vomiting and anorexia degree in treatment group from the day of chemotherapy to the 3 days after chemotherapy were significantly super to those of control group (P < 0. 05). Conclusions Improved TCM hot ironing technology can improve gastrointestinal reaction through chemotherapy period and worthy for clinical promotion.
8.Effect of CTP guided thrombolytic therapy in the treatment of acute cerebral infarction
Luyan GAO ; Hongxin WANG ; Peifen LIANG ; Yinhua DONG ; Lan ZHAO ; Jiangwei TANG ; Qiang LI ; Hongguang FAN ; Lina ZHOU
Tianjin Medical Journal 2015;(12):1437-1439
Objective To investigate the effect of CT perfusion (CTP) imaging guidance in the treatment of acute cere?bral infarction. Methods Patients (n=200) with acute cerebral infarction who visited our clinic within 6 hours underwent CTP examination and were divided into two groups:penumbra group and non-penumbra group according to their CTP imag?ing (presence of penumbra or not). Recombinant tissue plasminogen activator (rt- PA) was administrated for intravenous thrombolysis in both groups. NIHSS (The NIH Stroke Scale), BI (Barthel Index), mRS (modified Rankin Scores) and hemor?rhagic transformation events of two groups were determined before and after thrombolysis to evaluate its effect and prognosis in these two group. Results Compared with non penumbra group, NIHSS was reduced in penumbra group from 7 days after rt-PA (6.67±3.46 vs 4.76±2.04), and this decrease became obvious at 4 weeks after rt-PA (6.67±3.46 vs 3.68±1.93). Effi?ciency rate at 4 week (60.3%) and good prognosis rate at 3 months(71.7%)were both significantly improved in penumbra group than those in non penumbra group(34.7%,56.8%). Conclusion rt-PA under CTP guidance is effective and safe in the treatment of acute cerebral infarction. The thrombolytic therapy window can be enlarged according to the presence of pen?umbra or not and the bleeding conversion rate remains at low level.
9.Assay of adiponectin, leptin, true insulin and ghrelin levels in preterm human milk, and its relationship with infants growth.
Luyan HAN ; Ming LI ; Xinting YU ; Xiujing SUN ; Danhua WANG
Chinese Journal of Pediatrics 2014;52(7):510-515
OBJECTIVETo understand adiponectin, leptin, insulin and ghrelin levels in preterm colostrum and mature milk and their influence on the growth and development of the premature infant.
METHODThe study subjects were divided into two groups: preterm group and control group. Specimens of colostrum and mature milk on 42nd day after delivery were collected, the general situation of maternal and infants growth parameters at birth and at postnatal 42 days were recorded. Leptin, adiponectin, insulin and ghrelin levels in colustrum and mature milk were determined and compared.
RESULTA total of 128 mother-infant pairs were involved. There were 128 specimens of colostrums (80 from preterm group, 48 from control group) and 94 specimens of mature milk(50 from premature group, 44 from control group). The levels of colostrum, mature milk adiponectin, leptin, and insulin were not significantly different between the 2 groups; ghrelin levels in colostrum and mature milk of premature group were significantly lower than those in control group (P = 0.038), adiponectin and leptin levels in colostrum were higher than those of the mature milk (P < 0.05), colostrum ghrelin levels were lower than those of mature milk (P < 0.05). Adiponectin, leptin, and ghrelin showed no significant difference between different gestational age groups ( ≤ 34 weeks group vs. > 34 weeks group). True insulin level of mature milk in 34 weeks group was higher than that of > 34 weeks group (29.3 vs. 21.6 mU/L, P = 0.045); true insulin level in colostrums in ≤ 34 weeks group was lower than that in mature milk (21.7 vs. 29.3 mU/L, P = 0.000). Adiponectin levels in colostrum and 42 days weight gain were negatively correlated (r = -0.362, P = 0.025) . Insulin level in mature milk had a negative correlation with birth weight (r = -0.319, P = 0.029) . Ghrelin levels in colostrum and birth weight, length, head circumference, head circumference on 42(nd) day were positively correlated (r = 0.271,0.261,0.360, P < 0.05); weight, length at 42(nd) day and ghrelin levels showed borderline positive correlation (P = 0.050, 0.058).
CONCLUSIONMany bioactive hormones in milk might participate in the regulation of suitable growth after birth. Premature birth affects hormone levels in breast milk. Breast feeding is very important in preterm infants.
Adiponectin ; analysis ; Birth Weight ; physiology ; Breast Feeding ; Colostrum ; chemistry ; Female ; Gestational Age ; Ghrelin ; analysis ; Humans ; Infant ; Infant Nutritional Physiological Phenomena ; Infant, Newborn ; Infant, Premature ; growth & development ; Insulin ; analysis ; Leptin ; analysis ; Male ; Milk, Human ; chemistry ; Weight Gain ; physiology
10.Construction and validation of a dual-luciferase reporter gene system for screening and evaluating anti-liver fibrosis drugs that inhibit transcription of the gene encoding collagen I, chain a1.
Wei ZHANG ; Xiaoming DAI ; Hong YU ; Luyan WANG ; Shihui SUN ; Junfeng LI ; Yusen ZHOU
Chinese Journal of Hepatology 2014;22(10):747-751
OBJECTIVETo construct a dual-reporter gene system that will be applicable for use as a tool to screen and evaluate therapeutic drug compounds that inhibit transcription of the gene encoding collagen I, chain at1 (COL1A1).
METHODSThe full-length eDNA of transforming growth factor beta1 (TGFbeta1) was cloned by RT-PCR and inserted into two vectors, pcDNA3.1 and pJW4303, for construction of two eukaryotic expression vectors, pcDNA3.1-TGFbeta1 and pJW4303-TGFbeta1.Next, the promoter region of COL1A1, cloned by PCR using human genome DNA as template, was inserted into the vector pGL4.29 to construct the reporter gene vector, pGL4.29-COL1A1 promoter.All three recombinant vectors were verified by restriction enzyme digestion and DNA sequencing.Either the pcDNA3.1-TGFbeta1 or pJW4303-TGFbeta1 vector along with the pGL4.29-COL1A1 promoter vector or a pRL-null, control reporter, vector were co-transfected into the LX-2 human hepatic stellate cells to establish the transcription-activated dualreporter gene system.This system was used as a cell model for screening anti-liver fibrosis compounds that inhibit the transcription of COL1A1.Dexamethasone, a model drug that is known to inhibit the expression of COL1A1, was used as a control to validate the dual-reporter gene system.
RESULTSThe two TGFbeta1-expressing vectors and the reporter gene vector containing the promoter region of COL1A1 were successfully constructed.The results of a dual-reporter gene assay showed that TGFbeta1 co-expression increased the activity of the COL1A1 promoter by above 200-fold (t =21.78, P =0.0001), whereas in the absence of TGF31 co-expression the activity was below 2-fold (t =3.396, P =0.0274).The transcriptionactivated dual-reporter gene system was successfully established.The model drug, dexamethasone, effectively inhibited the activity of the COL 1A1 promoter in dose-dependent manner; the activity decreased 29.6% with 10 mumol/L dexamethasone (t =4.140, P =0.0144) and 53.9% with 100 mumol/L (t =6.193, P =0.0035).
CONCLUSIONThe dual-luciferase reporter system of TGFbeta1 and COL1A1 co-expression developed here can be used as a cell model to screen and evaluate anti-liver fibrosis compounds that inhibit activity of the COL1A1.
Base Sequence ; Collagen Type I ; genetics ; Drug Evaluation, Preclinical ; Genes, Reporter ; Genetic Vectors ; Humans ; Liver Cirrhosis ; drug therapy ; Luciferases ; Promoter Regions, Genetic ; Transcriptional Activation ; drug effects ; Transfection ; Transforming Growth Factor beta1

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