Objective To investigate the distribution and antibiotic resistance of the enteric pathogens in Shanxi province to guide the choice of antibacterials.Methods 798 cases of fecal samples from patients with diarrhea were treated in the outpatient and inpatient.The suspected pathogen was separated then PCR,biochemical test,serum agglutination tests were used for identification of suspected pathogen,and bacterial pathogens constitution and patho-genic characteristics were analyzed.Antimicrobial susceptibility testing was conducted by diskdiffusion method for the suspected pathogen with 6 antimicrobial agents.Results 81 strains isolated from 798 specimens were positive with 10.15% for pathogen detection.Diarrheagenic Escherichia coli was the first frequently pathogen,accounting for 46.91%,followed by Shigella,Salmonella and Aeromonas.Drug sensitivity monitoring showed that the most of the 81 strains had lower level of resistance to cefoitin,cefotaxime and ciorofloxacin,and higher level of resistance to tetra-cycline and nalidixic aid.Conclusion Diarrheagenic Escherichia coli,Shigella,Salmonella and Aeromonas are the major bacterial diarrheal pathogens in the hospital.Surveillance of antimicrobial resistance in these bacteria should be strengthened to provide reliable evidence for clinical anti infection treatment.

Objective To construct a shRNA lentiviral vector targeting the gene encoding tumor necrosis factor alpha-induced protein 8 (TNFAIP8) in RAW264. 7 cells, a mouse macrophage cell line, and to investigate the effects of TNFAIP8 gene silencing on the functions of mouse macrophages. Methods The shRNA sequence targeting TNFAIP8 gene was designed and DNA oligos containing small hairpin frame was synthesized. The double-stranded DNA was cloned into pLKO. 1-TRC vector after annealing. The recombi-nant vector was verified by using double enzyme digestion and gene sequencing. Lentiviruses were prepared by transfecting the constructed vector into 293T cells. Fluorescent quantitative RT-PCR and Western blot as-say were performed to detect the expression of TNFAIP8 at mRNA and protein levels after infecting the RAW264. 7 cells with lentiviruses. Flat dish adhesion experiment and wound-healing assay were used to evaluate the effects of TNFAIP8 gene silencing on the adhesion and migration of RAW264. 7 cells. Results The recombinant lentiviral vector was successfully constructed as indicated by double enzyme di-gestion and gene sequencing analysis. The expression of TNFAIP8 in RAW264. 7 cells at both mRNA and protein levels were significantly down-regulated after lentivirus infection (P<0. 05). Moreover, TNFAIP8 gene silencing significantly impaired the cell adhesion ability of RAW264. 7 cells after 15 min, 30 min or 2 hours of culture. Compared with the cells in control group, the RAW264. 7 cells harboring silenced TN-FAIP8 gene looked round with a smaller number of cellular extensions. The wound-healing assay showed that less TNFAIP8 gene-silenced RAW264. 7 cells migrated into the wounded area as compared with the cells in control group after 24 hours of culture (P<0. 05). The wound-healing rates of the experimental and control groups were 25% and 50%, respectively. Conclusion The recombinant lentiviral vector containing shRNA targeting the TNFAIP8 gene was successfully constructed. Transfecting the RAW264. 7 cells with the con-structed vector significantly silenced the expression of TNFAIP8 gene and inhibited the adhesion and migra-tion of these cells.

Objective To compared two classical rat models of nephrotic syndrome and to provide some reference data to researchers.Methods Thirty male SD rats were randomly divided into control group,puromycin aminonucleoside-induced nephrotic syndrome (PAN) group and adriamycininduced nephrotic syndrome (ADR) group.The body weight,twenty four hour proteinuria level,serum albumin concentration,cholesterol concentration,creatinine and urea concentration were measured.The renal pathology change was evaluated.The drug toxic effects,administration methods and the costs were also compared.Results There was no significant difference in body weight and hair color between control group and PAN group.Compared to control group,the body weight of the rats significantly decreased at day 15 and day 21 in ADR group (P ＜ 0.01),accompanied by epilation and diarrhea.Compared to control group,the 24-hour urinary protein levels increased significantly at day 10 (P ＜ 0.01),day 15 (P ＜ 0.01),and reached the peak level at day 15 (P ＜ 0.01),day 21 (P ＜ 0.01) in PAN group and ADR group respectively.Compared to control group,the serum albumin concentration decreased significantly at day 10 (P＜0.01),and return to normal level at day 15.The serum cholesterol concentration was increased significantly at day 10 (P ＜ 0.01) and return to normal at day 15 in PAN group.Compared to control group,the serum albumin concentration was decreased significantly at day 15 (P＜0.05) and return to normal at day 21 in ADR group.No significant difference of serum creatinine and serum urea nitrogen levels were found among three groups.Compared to control group,the width of foot process increased significantly at day10 (P ＜ 0.01) and day 15 (P ＜ 0.05) in PAN group and ADR group respectively.To successfully induce a nephrotic rat model (per 100 g),the cost of PAN group was 3.1 times of ADR group (578.10 yuan vs 186.94 yuan).Conclusions Nephrotic syndrome can be induced by both PAN and ADR.The administration of PAN via intraperitoneal injection is more convenient as compared to ADR via tail intravenous injection.Compared to ADR,PAN can induce nephrotic syndrome model more rapidly,with more consistent detection index,and less toxic effects,but its cost is more expensive.

Objective: To investigate the effect of losartan on angiotensin II (Ang II) expression and myocardial remodeling in myocardial infarction (MI) rats’ model.
Methods: A total of 32 SD male rats were divided into 4 groups, Sham operation group, MI group, MI with losartan 10mg/(kg·d) group and MI with losartan 20mg/(kg·d). n=8 in each group. MI model was established and the electrocardiogram changes before and after MI were recorded, hemodynamic indexes were detected at 4 weeks after MI, pathological changes of myocardial tissue were examined by HE staining. The myocardial mRNA and protein expressions of ACE2 and Ang II were detected by RT-PCR and Western Blot analysis.
Results: Compared with Sham operation group, MI group showed increased LVMI and decreased LVEF P<0.05;the above changes were getting better in both MI with losartan groups in a dose-dependent manner. The pathological examination presented that MI group had myocardial cell swelling, fracture, hyperplasia and inflammatory cell infiltration, those damages were less in MI with losartan groups in a dose-dependent manner, Sham operation group had no pathological changes. Compared with Sham operation group, the mRNA and protein expressions of Ang II were obviously higher in MI group, P<0.05 and the expressions were decreased in MI with losartan groups in a dose-dependent manner;the mRNA and protein expressions of ACE2 were slightly increased in MI group and the expressions were further increased in MI with losartan groups in a dose-dependent manner.
Conclusion: Losartan could increase ACE2 expression and therefore, inhibit Ang II expression and improve the ventricular remodeling in MI rats’ model.

Objective To investigate the expression and aberrant methylation of Ptchl gene in hedgehog signal pathway in carcinogenesis of human gastric cancer.Methods The total RNA and genomic DNA were extracted from 10 human gastric carcinoma tissues,adjacent tissues(＞3 cm from cancerous tissue)and gastric cancer eell line AGS.Ptchl mRNA expression was detected by real-time quantitative reverse transcription PCR.The pattern of CpG island in Ptchl gene 5'regulation sequence was analyzed by software and its methylation extent was tested by bisulfite sequencing PCR.Results The analysis of CpG island(starting-3950 bases upstream of the Ptchl mRNAla transcription start site and ending 2050 bases downstream)revealed that there were two CpG islands in Ptchl gene 5' regulation sequence(first CpG:-1139 bp～+860 bp;second CpG:+875 bp～+1692 bp).Bisulfite sequencing PCR analysis of 19 CpG sites included in the first CpG island(-870 bp～+229 bp)showed that there was methylation present in all cell lines and the average extent of the methylation of these CpG sites was significantly higher in cancerous tissues(64%±32%,ranged 16%～100%)than that in adjacent tissues(13%±14%,ranged 0%～42%,P＜0.05).There was a negative correlation of the Ptchl methylation with its expression.Conclusion The high methylation of Ptchl gene that involves in the carcinogenesis of human gastric carcer will be a new biomarker for gastric carcer.

Objective To develop a tetra-primer amplification refractory mutation system PCR(T-ARMS-PCR)assay for detecting four single nucleotide polymorphisms(SNPs): rs1801133,rs1801131, rs1805087 and rs1801394 associated with folate metabolism in a single reaction tube.Methods Methodology was developed.Then, it applied the established method to analyze 150 physical examination children′s anticoagulant blood samples admitted to the Department of Clinical Laboratory, Children′s Hospital of Hebei Province from January 2017 to April 2017.Four sets of chimeric primers consisting of a universal Tag sequence and targeting rs 1801133,rs1801131,rs1805087 and rs1801394 fused to the specific sequence were designed according to the T-ARMS-PCR principle and the multiplex chimeric primers strategy.A single tube PCR was then conducted after optimizing the primer concentrations and the reaction conditions.The amplified products were analyzed by QIAxcel capillary electrophoresis, and the corresponding SNP genotypes of 150 samples were identified.Furthermore,all samples were verified by direct sequencing.And the Hardy-Weinberg Equilibrium(HWE)testing of four SNPs from 150 samples were conducted by SPSS17.0 Chi-square test.Results The improved T-ARMS-PCR combined with capillary electrophoresis can accurately verify eight different alleles of the four SNPs associated with folate metabolism at one time in 3 hours.Four SNPs of 150 whole blood samples were accurately classified and the results were completely consistent with direct sequencing.All the genotype frequencies of these four SNPs were in HWE (χ2rs1801133=0.69, Prs1801133=0.40; χ2rs1801131=0.21, Prs1801131=0.64; χ2rs1805087=3.32, Prs1805087=0.07;χ2rs1801394=1.91, Prs1801394=0.17).Conclusions The proposed improved T-ARMS-PCR combined with capillary electrophoresis in this study can accurately verify four SNPs associated with folate metabolism in a single reaction tube.This method might be a valuable tool to specifically guide the folate supplement in general population.

Objective:To explore the expression and clinical significance of S100A8 and S100A9 in Helicobacter pylori ( H. pylori) associated gastritis. Methods:A total of 101 patients with chronic gastritis diagnosed in the First Hospital of Shanxi Medical University from October 2018 to May 2019 were selected. The expression levels of S100A8 and S100A9 in the gastric mucosa tissues of 101 patients with chronic gastritis were determined by immunohistochemistry (in absorbance), and the mRNA expression levels of S100 A8 and S100 A9 in the gastric mucosa tissues of 48 patients were detected by reverse transcription-polymerase chain reaction. And the results combined with pathological diagnosis of routine staining and clinical H. pylori infection data were analyzed. Mann-Whitney U test, Kruskal-Wallis H test and Spearman rank correlation were used for statistical analysis. Results:Among 101 patients, there were 59 cases of chronic atrophic gastritis (CAG group) and 42 cases of chronic non-atrophic gastritis (NAG group); 59 cases were H. pylori positive ( H. pylori positive group) and 42 cases were H. pylori negative ( H. pylori negative group). There were statistically significant differences in the expression levels of S100A8 and S100A9 between CAG group and NAG group (0.10, 0.07 to 0.13 vs. 0.09, 0.06 to 0.10 and 0.13, 0.08 to 0.15 vs. 0.09, 0.07 to 0.10, respectively), and between H. pylori positive group and H. pylori negative group (0.11, 0.10 to 0.13 vs. 0.07, 0.06 to 0.08 and 0.13, 0.10 to 0.15 vs. 0.07, 0.07 to 0.08, respectively) ( U=754.00, 602.00, 5.00 and 40.00, all P<0.01). There were statistically significant differences in the expression levels of S100A8 and S100A9 between H. pylori positive patients (34 cases) and H. pylori negative patients (25 cases) in CAG group (0.13, 0.11 to 0.14 vs. 0.07, 0.07 to 0.08 and 0.15, 0.14 to 0.16 vs. 0.08, 0.08 to 0.09, respectively), similarly, there were significant differences in the expression levels of S100A8 and S100A9 between H. pylori positive patients (25 cases) and H. pylori negative patients (17 cases) in NAG group (0.10, 0.09 to 0.10 vs. 0.06, 0.05 to 0.07 and 0.10, 0.10 to 0.11 vs. 0.07, 0.06 to 0.07, respectively) ( U=1.00, 0.00, 0.00 and 0.00, all P<0.01). The results indicated that the expression levels of S100A8 and S100A9 were high in H. pylori positive patients in CAG group, the expression levels of S100A8 and S100A9 were low in H. pylori negative patients in NAG group, and the differences were statistically significant ( H=84.78 and 89.64, both P<0.01). There were statistically significant differences in the expression of S100 A8 and S100 A9 at mRNA level between CAG group (24 cases) and NAG group (24 cases) (0.12, 0.06 to 1.31 vs. 0.05, 0.03 to 0.08; 0.19, 0.03 to 0.43 vs. 0.03, 0.01 to 0.09), and the expression of S100 A8 and S100 A9 at mRNA level was significant between H. pylori positive patients (24 cases) and H. pylori negative patients (24 patients) (0.45, 0.10 to 1.90 vs. 0.05, 0.03 to 0.08 and 0.36, 0.24 to 0.81 vs. 0.03, 0.01 to 0.04) ( U=55.00, 74.00, 19.00 and 2.00, all P<0.05). There were statistically significant differences in the expression of S100 A8 and S100 A9 at mRNA level between H. pylori positive patients (12 cases) and H. pylori negative patients (12 cases) of CAG group (0.85, 0.27 to 2.28 vs. 0.06, 0.03 to 0.09 and 0.39, 0.25 to 0.87 vs. 0.03, 0.02 to 0.05), and the expression of S100 A8 and S100 A9 at mRNA level was significant between H. pylori positive patients (12 cases) and H. pylori negative patients (12 cases) of NAG group (0.09, 0.05 to 0.28 vs. 0.04, 0.03 to 0.07 and 0.20, 0.09 to 0.65 vs. 0.01, 0.01 to 0.03) ( U=5.00, 2.00, 0.00 and 0.00, all P<0.01). The results showed that the expression of S100 A8 and S100 A9 at mRNA level was high in H. pylori positive patients in CAG group, the expression of S100 A8 and S100 A9 at mRNA level was low in H. pylori negative patients in NAG group, and the differences were statistically significant ( H=20.43 and 24.15, both P<0.01). The expression levels of S100A8 and S100A9 were positively correlated at both protein level and mRNA level ( r=0.899 and 0.903, both P<0.01). Conclusions:S100A8 and S100A9 may involve in the inflammation process of H. pylori-infected gastric mucosa and promote the proliferation of gastric epithelial cells, which may be one of mechanisms of intrinsic glands reduction and CAG genesis. S100A8 and S100A9 are expected to be potential biomarkers for diagnosis and follow-up and potential targets for treatmert of CAG.

Objective To explore the application effects of evidence-based nursing in percutaneous endoscopic gastrostomy ( PEG ) postoperative enteral nutrition and the effects of prevention and treatment of complications. Methods A retrospective and prospective cohort study method was adopted. The control group was composed of 58 cases of PEG enteral nutrition patients, on whom evidence-based nursing was not conducted before January 2013, while the intervention group was composed of 60 cases of PEG enteral nutrition patients, on whom evidence-based nursing was conducted after January 2013. Intervention effects of nursing were compared and analyzed between two groups. Results The incidence of complications in the intervention group (11. 67%) was significantly lower than that of the control group (43. 10%) (P<0. 01), and the incidence of infectious complications in the intervention group (5. 00%) was significantly lower than that of the control group (20.69%) (P<0. 01). There was no significant association between feeding time and complications (P >0. 05);there was significant association between ways of nutrition infusion and replacement rate of skin dressing (P<0. 01). Conclusions Application of evidence-based nursing in postoperative enteral nutrition of PEG patients can effectively prevent complications from happening, promote recovery of patients′ gastrointestinal function and overall nutritional status, and eventually shorten the time of hospitalization.

OBJECTIVETo investigate RANK-RANKL expression in the kidneys of a rat model of puromycin aminonucleoside nephropathy (PAN).

METHODSThirty-six SD rats were randomly divided into PAN model group and normal control group. PAN was induced by a single intravenous injection of 100 mg/kg puromycin aminonucleoside. Serum creatinine and 24-hour urinary protein were measured on days 3, 7, and 14 after the injection, and renal pathologies were assessed with optical and immune transmission electron microscopy. The expression of RANK and RANKL in the kidneys was examined using reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting.

RESULTSThe PAN model rats showed massive proteinuria and elevated serum creatinine on day 3, which peaked on day 7. RANK-RANKL protein and mRNA expressions in PAN model group was higher than those in the control group. In the PAN rats, RANK was expressed mainly on the top cell membrane and in the cytoplasm of renal podocytes with a significantly increased expression level compared with that in the control group.

CONCLUSIONThe PAN rat model shows aberrant RANK and RANKL expressions in the podocytes, indicating their contribution to podocyte injury in PAN.

Animals ; Creatinine ; blood ; Female ; Kidney ; drug effects ; metabolism ; Kidney Diseases ; chemically induced ; metabolism ; pathology ; Male ; Podocytes ; drug effects ; metabolism ; Proteinuria ; pathology ; Puromycin Aminonucleoside ; adverse effects ; RANK Ligand ; metabolism ; Rats ; Rats, Sprague-Dawley ; Receptor Activator of Nuclear Factor-kappa B ; metabolism

Objective To investigate RANK-RANKL expression in the kidneys of a rat model of puromycin aminonucleoside nephropathy (PAN). Methods Thirty-six SD rats were randomly divided into PAN model group and normal control group. PAN was induced by a single intravenous injection of 100 mg/kg puromycin aminonucleoside. Serum creatinine and 24-hour urinary protein were measured on days 3, 7, and 14 after the injection, and renal pathologies were assessed with optical and immune transmission electron microscopy. The expression of RANK and RANKL in the kidneys was examined using reverse transcription-ploymerase chain reaction (RT-PCR) and Western blotting. Results The PAN model rats showed massive proteinuria and elevated serum creatinine on day 3, which peaked on day 7. RANK-RANKL protein and mRNA expressions in PAN model group was higher than those in the control group. In the PAN rats, RANK was expressed mainly on the top cell membrane and in the cytoplasm of renal podocytes with a significantly increased expression level compared with that in the control group. Conclusion The PAN rat model shows aberrant RANK and RANKL expressions in the podocytes, indicating their contribution to podocyte injury in PAN.