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Objective:To investigate the efficacy of metagenomic next generation sequencing (mNGS) in the etiological diagnosis of patients with spinal infection, so as to provide reference for timely diagnosis and treatment.Methods:A total of 40 patients with suspected spinal infection admitted to the Department of Infectious Diseases in Henan Provincial People′s Hospital from January 2020 to July 2022 were included. The results of tissue culture, histopathological examination and tissue mNGS detection were analyzed retrospectively. According to the clinical diagnose, the patients were divided into the spinal infection group (28 cases) and the non-spinal infection group (12 cases). The positive rate, sensitivity and specificity of mNGS and tissue culture in the pathogen detection of patients with spinal infection were compared. McNemar test was used for statistical analysis.Results:There were 23 males and 17 females in 40 patients. The positive rate of mNGS was higher than that of tissue culture (75.0%(30/40) vs 12.5%(5/40)), and the difference was statistically significant ( χ2=0.08, P<0.001). Based on clinical diagnostic criteria, the sensitivity of mNGS in the diagnosis of spinal infection was higher than that of tissue culture (82.1% vs 17.9%), with a statistically significant difference ( χ2=0.02, P<0.001), while the specificity compared to the tissue culture (33.3% vs 100.0%), the difference was not statistically significant ( P>0.05). Conclusions:mNGS has a high pathogen detection rate and sensitivity in the etiological diagnosis of patients with spinal infection, which could provide clinical guidance for the diagnosis and treatment of patients with spinal infection.
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Objective:To analyze the liver pathological characteristics of chronic hepatitis B (CHB) patients with normal alanine aminotransferase (ALT) and negative hepatitis B e antigen (HBeAg), and to evaluate the diagnostic value of different serological models for liver fibrosis.Methods:Retrospective analysis was conducted on the patients with HBeAg-negative CHB who had normal ALT and underwent liver biopsy from August 2016 to December 2019 in the Department of Infectious Diseases, Henan Provincial People′s Hospital. The clinical data, serum indicators of hepatitis B virus (HBV) and HBV DNA were collected. The liver fibrosis stages (S) was assessed by pathological examination. The diagnostic efficacies of gamma-glutamyl transpeptidase to platelet ratio (GPR), fibrosis 4 score (FIB-4), S index, aspartate aminotransferase to platelet ratio index (APRI) and gamma-glutamyl transpeptidase to albumin ratio (γ-GT/ALB) for liver pathological fibrosis were analyzed by the receiver operating characteristic curves. Two variable correlation test was used to explore the relationship between the different models and pathological fibrosis of liver tissue. Chi-square test was used for statistical comparison.Results:The age of 448 patients was (37.98±9.82) years, and the male to female ratio was 1.286 ∶1. The proportions of S≥2 in patients with age>30 years, hepatitis B surface antigen (HBsAg)<2 000 IU/mL and HBV DNA≥2 000 IU/mL were higher than those in patients with age ≤30 years, HBsAg ≥2 000 IU/mL and HBV DNA<2 000 IU/mL, respectively, and the differences were all statistically significant ( χ2=7.68, P=0.006; χ2=11.44, P=0.001; χ2=9.12, P=0.003, respectively). There were 250 cases with pathological fibrosis stage S<2, 162 cases with S=2 and 36 cases with S≥3. FIB-4 (correlation coefficient 0.250), APRI (correlation coefficient 0.218), GPR (correlation coefficient 0.186), S index (correlation coefficient 0.184) and γ-GT/ALB (correlation coefficient 0.127) were positively correlated with the severity of liver fibrosis (all P<0.050). S index had the highest sensitivity (64.1%) in the diagnosis of significant liver fibrosis (S≥2), while γ-GT/ALB had the highest specificity (80.8%). In the diagnosis of severe liver fibrosis (S≥3), γ-GT/ALB had the highest sensitivity (77.8%), while APRI had the highest specificity (78.6%). Conclusions:The incidence of liver fibrosis in CHB patients with normal ALT and negative HBeAg is relatively high. The current serological diagnostic models are not suitable for the evaluation of liver fibrosis in these patients, and timely liver puncture is still necessary.
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Objective:To explore the effects of small-incision carpal tunnel release on postoperative functional recovery and electrophysiological indexes in carpal tunnel syndrome (CTS).Methods:A total of 75 patients with CTS treated in Shulan (Hangzhou) Hospital were enrolled as the research objects between April 2016 and April 2021. According to different surgical methods, they were divided into group A (34 cases, small-incision carpal tunnel release) and group B (41 cases, traditional carpal tunnel release). The operation time, hospitalization time, time to resume work, electrophysiological indexesbefore and after surgery, and postoperative complications were compared between the two groups.Results:Compared with group B, operation time, hospitalization time and time to resume work were shorter in group A: (12.32 ± 3.26) min vs. (34.65 ± 7.49) min, (5.15 ± 1.68) d vs. (7.83 ± 2.24) d, (18.22 ± 2.03) d vs. (37.35 ± 3.16) d ( P<0.05). After surgery, electrophysiological indexes in both groups were improved ( P<0.05), but there was no significant difference between the two groups ( P>0.05). The incidence rates of scar pain, decreased grip strength and hand piercing pain in group A were lower than those in group B: 2.94%(1/34) vs. 19.51%(8/41), 0 vs. 21.95%(9/41), 0 vs. 12.20%(5/41) ( P<0.05). Conclusions:Compared with traditional carpal tunnel release, clinical curative effect of small-incision carpal tunnel release is comparable on CTS patients. However, it can shorten operation time, hospitalization time and time to resume work, reduce incidence of postoperative complications.
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Objective To investigate whether there are differences in lymphocyte subsets between chronic hepatitis B (CHB) patients receiving different antiviral treatment regimens, and to determine related predictive factors for HBsAg decline. Methods A retrospective analysis was performed for 68 treatment-experienced CHB patients who attended the outpatient service in Department of Infectious Diseases, Henan Provincial People's Hospital, from October to December 2019, and according to the antiviral treatment regimen, they were divided into PEG-IFNα treatment group with 10 patients, PEG-IFNα+nucleos(t)ide analogues (NAs) treatment group with 21 patients, and NAs treatment group with 37 patients. Related data were recorded, including demographic features, blood routine, albumin, HBsAg, and measurement of lymphocyte subsets. A one-way analysis of variance was used for comparison of normally distributed continuous data between groups, and the Kruskal-Wallis H test was used for comparison of non-normally distributed continuous data between groups; the chi-square test was used for comparison of categorical data between groups; the multivariate logistic regression analysis was used to investigate independent influencing factors for HBsAg decline. Results There were significant differences between the three groups in HBsAg decline ( H =8.348, P =0.015), absolute value of lymphocytes ( F =4.643, P =0.013), and T lymphocyte count ( F =7.721, P =0.001). The multivariate logistic regression analysis showed that sex (odds ratio [ OR ]=0.227, 95% confidence interval [ CI ]: 0.059-0.878, P =0.032), age ( OR =0.931, 95% CI : 0.868-0.999, P =0.047), antiviral treatment regimen (PEG-IFN-α treatment group vs NAs treatment group: OR =9.600, 95% CI : 1.982-46.498, P =0.005; PEG-IFN-α+NAs treatment group vs NAs treatment group: OR =4.800, 95% CI : 1.336-17.243, P =0.016), and T lymphocyte count ( OR =0.804, 95% CI : 0.684-0.944, P =0.008) were independent influencing factors for HBsAg decline. Conclusion For CHB patients receiving PEG-IFNα alone or in combination with NAs, monitoring of lymphocyte subsets during the treatment process may help to judge HBsAg decline, and the lower the absolute value of T lymphocytes, the greater the possibility of HBsAg decline.
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ObjectiveTo investigate the status of abnormal renal function markers and related influencing factors in chronic hepatitis B (CHB) patients receiving oral antiviral drugs for a long time. MethodsA retrospective analysis was performed for the clinical data of 681 CHB patients who attended Henan Provincial People’s Hospital from January to December 2019 and received long-term oral administration of entecavir (ETV)/tenofovir disoproxil fumarate (TDF). All patients received the measurement of blood renal function markers (urea, creatinine, retinol-binding protein ［RBP］, cystatin C ［Cys-C］, and β2-microglobulin ［β2-MG］), urinary renal function markers (α1-microglobulin ［α1-MG］, Cys-C, and N-acetyl-β-D-glucosaminidase ［NAG］), and urine routine parameters. The incidence rate of abnormal renal function markers were analyzed. The McNemar’s test was used for comparison of categorical data between groups, and the multivariate logistic regression analysis was used to investigate independent influencing factors for abnormal renal markers in urine. ResultsThe 681 patients had a mean age of 39.8±11.0 years and received medication for 1.88 (0.80-3.16) years. There were 417 male patients and 264 female patients, and the incidence rate of liver cirrhosis was 27.02% (184/681). Of all 681 patients, 442 received ETV and 239 received TDF. The measurement of blood renal function markers showed that urea, creatinine, retinol-binding protein, Cys-C, and β2-MG had an abnormal rate of 6.9% (47/681), 0.15% (1/681), 0(0/681), 2.21% (15/681), and 5.03% (30/681), respectively, and the abnormal rate of urinary protein was 7.29% (49/672). The measurement of urinary renal function markers showed that α1-MG, NAG, and Cys-C had an abnormal rate of 38.62% (263/681), 37.74% (257/681), and 19.38% (132/681), respectively. The abnormal rate of urine test was higher than that of blood test (P＜0.001). The multivariate logistic regression analysis with urinary α1-MG as the dependent variable showed that sex (odds ratio ［OR］=0.293, 95% confidence interval ［CI］: 0.204-0.419, P＜0.05), age (OR=1298, 95%CI: 1.108-1.521, P＜0.05), and type of nucleoside drug (OR=2.100, 95%CI: 1.431-3.083, P＜0.05) were influencing factors. The multivariate logistic regression analysis with urinary NAG as the dependent variable showed that age (OR=1.177, 95%CI: 1.008-1.375, P=0.040) was an influencing factor. ConclusionCompared with blood renal function markers, urinary renal function markers can identify renal injury earlier in CHB patients, and the elderly patients and the patients receiving TDF are more likely to develop abnormal renal function. However, it is not observed whether the duration of medication and liver cirrhosis can increase the risk of renal injury in CHB patients.
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Objective:To evaluate the efficacy and safety of elbasvir/grazoprevir (EBR/GZR) in patients with genotype 1 chronic hepatitis C in the real-world.Methods:This was an open-label, single-center, retrospective real-world study. A total of 103 genotype 1 chronic hepatitis C patients who were treated with EBR/GZR in Henan Provincial People′s Hospital from May 2018 to October 2019 were enrolled.And the clinical baseline characteristics of patients and the effectiveness and safety of antiviral therapy were respectively evaluated.Results:A total of 103 patients were enrolled in the study with an age of (47.6±13.9) years. Fifty-five (53.4%) patients were male and 48(46.6%) were female. One point nine percent (2/103) patients were genotype 1a hepatitis C and 98.1%(101/103) were genotype 1b hepatitis C. Seventeen genotype 1b hepatitis C patients were previously treated with interferon, and three patients co-infected with hepatitis B virus (HBV). Among the 103 cases, 35 had underlying diseases and 26 had combined medication. Ninty-eight cases completed 12-week treatment and 89 cases completed 12-week follow-up after treatment.Overall, 89 cases achieved sustained virological response. The overall incidence of adverse reactions was 20.4%(21/103), and the main adverse reactions were fatigue, insomnia and anxiety. No serious adverse event occurred. The three patients with HBV co-infection had no hepatitis B activation after treatment.Conclusion:EBR/GZR is effective and safe in the patients with genotype 1 chronic hepatitis C in China.
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Objective:To evaluate the efficacy of artificial liver blood purification system in the treatment of severe and critical patients with corona virus disease 2019(COVID-19), and to observe the dynamic changes of lymphocyte subsets and cytokines after treatment.Methods:A total of six patients with COVID-19 admitted to the ward of public health center of Henan Provincial People′s Hospital from January 31 to February 18, 2020 were enrolled, including three severe cases and three critical cases. The protocals of artificial liver treatment were developed according to the patients′ conditions, and the patients′ epidemiological history, clinical characteristics, and laboratory examination data were retrospectively analyzed. At the same time, dynamic changes of lymphocyte subsets and cytokines were detected before and after artificial liver treatment.Results:By February 24, 2020, two severe patients were discharged after cured, and one case was discharged after improved, with an average stay of 19 days. Two critical patients were still hospitalized and one died. Three severe patients were all treated with hemofiltration, while two critical patients were treated with hemofiltration plus plasma exchange, and one was treated with continuous bedside hemofiltration.Among six patients, the ratio of neutrophils to lymphocytes before treatment were 6.42, 2.63, 15.00, 15.09, 12.04 and 30.41, respectively. After treatment, the ratio of neutrophils to lymphocytes became 4.77, 6.05, 4.86, 5.43, 32.77 and 23.46, respectively.The absolute numbers of lymphocytes in six patients before treatment were low, with a median of 382/μL. After treatment of artificial liver, the absolute numbers of lymphocytes increased, with a median of 476/μL. Cytokines were detected in three critical patients, and the interleukin 6 (IL-6) levels in two cases were 26 042.00 ng/L and 282.03 ng/L, respectively before treatment. After treatment, the levels decreased to 226.85 ng/L and 26.15 ng/L, respectively. IL-6 continued increasing from 30.14 ng/L to 709.25 ng/L in one another critical patient, who eventually died.Conclusions:In severe and critical patients with COVID-19, artificial liver treatment can reduce inflammation and increase the absolute numbers of lymphocytes and the subsets. The IL-6 level may be correlated with disease progression and may be a useful prognostic factor for early identification of severe and critical COVID-19.
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Objective To explore the efficacy and safety of daclatasvir (DCV ) combined with asunprevir (ASV) for chronic genotype 1b (GT1b) hepatitis C .Methods Twenty-nine GT1b hepatitis C patients who were treated with DCV combined ASV in Henan Provincial People′s Hospital from September 2017 to November 2017 were included .Hepatitis C virus (HCV ) RNA levels were tested before treatment ,1 week ,2 weeks ,3 weeks ,4 weeks ,8 weeks ,12 weeks and 24 weeks after treatment , and 12 weeks after the end of the treatment .The comorbidities ,combined use of drugs and adverse clinical events were registered .T test was used to compare the measurement data with normal distribution and M (P25,P75) was used for measurement data with non-normal distribution .Results A total of 29 patients with GT1b were included ,with 4 cirrhosis cases and 25 non cirrhotic cases .Seven patients had history of previous interferon and ribavirin combination treatment .There were 9 patients with comorbidity and 7 patients with combined medication . Finally , 25 patients completed a 24-week course of antiviral treatment ;3 patients were lost to follow-up ,and 1 patient withdrew after 16weeks of antiviral treatment because of a virus rebound .Of the 26 followed up patients ,25 achieved sustained virological response at 12-week (SVR12 ) , and one patient failed .And the HCV RNA NS5A resistance-associated variants (RAV) were detected in the patients with treatment failure .No severe adverse clinical events occurred in 26 patients .Conclusions DCV combined with ASV is effective and safe in the treatment of GT 1b chronic hepatitis C .However , the effect of RAV on therapeutic efficacy should be concerned during the treatment .
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Objective@#To recognize the efficacy and safety of paritaprevir/ritonavir-ombitasvir combined with dasabuvir (OBV/PTV/RTV+DSV) in the treatment of genotype 1b chronic hepatitis C.@*Methods@#Patients with genotype 1b chronic hepatitis C who were admitted to the People's Hospital of Henan Province, Huashan Hospital of Shanghai and the Fifth Medical Center of the General Hospital of the People's Liberation Army of China between November 2017 to August 2018 were enlisted. All patients received OBV/PTV/RTV+DSV antiviral therapy. HCV RNA levels were measured at baseline, weeks 1, 2, 3, 4, 8, 12, and 24, then 12 weeks, and 24 weeks after completion of treatment; patients’ comorbidity, concomitant medications, and clinical adverse events were recorded.@*Results@#108 patients were enrolled in the study, with an average age of 49.1 years, 44 patients were male (40.8%), 96.3% (104/108) were newly diagnosed, and four patients had previous treatment history, of whom three were treated with IFN and one with IFN + DAA. Ninety-eight cases completed 12 weeks treatment and 89 cases were in follow up for 12 weeks, after discontinuation of the drug. Overall, 89 cases (100%) achieved SVR12.One patient treated with PR and DAA had HCV RNA level of 869175 IU/mL at 4 weeks of treatment, which was significantly higher than the baseline HCV RNA level (301776IU/ML), and was judged as failure of treatment; and follow-up was discontinued. Of all enrolled patients, 19 (17.6%) had underlying diseases and 15 (13.9%) had combined medications. During treatment, adverse events (AE) occurred in 11 patients (10.1%). The main adverse events were pruritus and elevated bilirubin.@*Conclusion@#Combined antiviral therapy (OBV/PTV/RTV+DSV) of 12 weeks are highly effective with good safety profile in the treatment of Chinese patients with genotype 1b chronic hepatitis C.
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Objective To analyze hepatitis C virus genotype(HCV GT)1b NS5A resistance-associated variants(RAV)and its related factors,and to provide references for direct-acting antivirals (DAA)agent selection and application.Methods From January 2017 to July 2017,53 hepatitis C patients were selected from the Department of Infectious Diseases of Henan Province People's Hospital. The mutations of L31M and Y93H in NS5A RAV were analyzed in 43 HCV GT1b patients,and their correlations with hepatitis C virus,liver function,platelet and liver fibrosis diagnostic model[APRI, gamma-glutamyl transpeptidase to platelet ratio(GPR),FIb-4]were analyzed.The quantitative data were compared by two independent samples t test,and the qualitative data were compared by chi square test. Results Fifty-three subjects were enrolled,including 43 GT1b(9 males and 34 females)and 10 GT2a(2 males and 8 females).No other genotype was detected.The incidence of NS5A RAV in 43 HCV GT1b patients was 13.9%(6/43),of which L31M and Y93H were 1/43(2.3%)and 5/43(11.6%)with no significant difference(χ2= 1.500,P= 0.219).There were no significant differences in HCV RNA, ALT,AST,albumin,platelets and age between patients with or without mutation(all P> 0.05). Conclusions The incidence of NS5A RAV in HCV GT1b patients is high,but not affected by virus, biochemical factors and liver fibrosis.The detection of NS5A RAV before HCV treatment is helpful for rational selection of DAA,which could reduce the drug resistance.
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AIM:To investigate the role of reactive oxygen species (ROS) in the regulation of intracellular Ca2+induced by angiotensin II ( Ang II) in the primarily cultured medullary neurons .METHODS:Primarily cultured me-dullary neurons were prepared from 14-day-old embryos of Sprague-Dawley rats in the study .The identification of medullary neurons was assessed by double-labeling immunofluorescence .To explore the role of ROS , mainly the superoxide ( O2 ·-) , the O2 ·-generation was measured using the fluorogenic probe dihydroethidium ( DHE) .To determine intracellular free cal-cium concentration ( [ Ca2+] i ) , the neurons were loaded with the Ca 2+-specific dye Fura-2/AM.The cell viability after adding Ang II was also examined using CCK-8 assay.RESULTS:Most of the cultured cells were medullary neurons , more than 80%of which were glutamate positive neurons .Ang II (5 μmol/L) increased the level of ROS within 10 min in the medullary neurons .Ang II at 5μmol/L induced a significant [ Ca2+] i increase in the medullary neurons , and the effect of Ang II occurred rapidly and reached a peak within 20 min after administration.The level of [Ca2+]i started to decline after washout .The Ca2+elevation induced by Ang II was significantly decreased by apocynin or TEMPOL .No significant differ-ence in the cell viability between control group and 5μmol/L Ang II treatment group was observed .CONCLUSION:ROS is involved in the regulation of [Ca2+]i induced by Ang II in the primarily cultured medullary neurons , suggesting a poten-tial intracellular signaling mechanism involved in the Ang II-mediated oxidant regulation of central neural control of blood pressure.
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<p><b>OBJECTIVE</b>To understand the distribution of diarrheagenic Escherichia (E.) coli in population in Shanghai and discuss the practice model of cooperation in enteric infectious disease prevention and control between public health institution and hospital.</p><p><b>METHODS</b>Sentinel hospitals were assigned, standard detection and identification of diarrheagenic E. coli were conducted, incidence curve of diarrheagenic E. coli infection was drawn and epidemiologic survey and laboratory detection were conducted for suspect diarrheagenic E. coli infection outbreaks.</p><p><b>RESULTS</b>A total of 7 204 stool specimens were collected from diarrhea patients in 4 hospitals during 2012-2013, in which 712 (9.9% ) were diarrheagenic E. coli positive, including 351 enteropathogenic E. coli (EPEC) strains, 292 enterotoxigenic E. coli (ETEC) strains, 32 enteroinvasive E. coli(EIEC) strains and 6 Shiga toxin-producing E. coli (STEC/EHEC) strains, as well as 31 mixed strains. EPEC infection mainly occurred in children aged 1-5 years; and all of these infections were caused by aEPEC. The incidence peak of ETEC infection was during August, the positive rate was >20%. The ETEC infection mainly occurred in infants aged 1-28 days in 2012 and in people aged 20-60 years in 2013 (P<0.05). ST was the major type (59.6%), followed by LT (27.8%) and ST/LT (12.6%). EIEC infection increased in children obviously in 2013 (P<0.01). No EHEC O157:H7 case was detected, but two EHEC O26:H11 (eae-hlyA-stx1a) cases in children were reported for the first time in Shanghai. The survey result indicated that the multidrug-resistant ETEC (STh-CS21-CFA/I-ClyA-EatA-ST2332-SHNL0005) strain causing outbreak in 15 newborns in Shanghai in 2012 was in the same clone as the strain detected in Zigong in Sichuan province.</p><p><b>CONCLUSION</b>Significant change has occurred in diarrheagenic E. coli distribution in Shanghai in recent years, ETEC has potential risk to cause outbreak of hospital acquired infection in neonates and food borne infection. The active surveillance on ETEC and other enteric pathogens by both public health institutions and hospitals need to be improved.</p>
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Adult , Child, Preschool , Humans , Infant , Infant, Newborn , Middle Aged , Young Adult , China , Epidemiology , Diarrhea , Microbiology , Disease Outbreaks , Enteropathogenic Escherichia coli , Enterotoxigenic Escherichia coli , Escherichia coli Infections , Epidemiology , Incidence , Sentinel SurveillanceABSTRACT
Objective To investigate the pathotypes,epidemiological characteristics and antimicrobial resistance of diarrheagenic Escherichia coli (DEC) in children with acute bacterial diarrhea in Shanghai.Methods A total of 2 071 outpatient children with probable acute bacterial diarrhea referred to the enteric clinic of Children's Hospital of Fudan University during June 2012 to June 2014 were enrolled in our study.The stool samples were processed for routine microbiologic and biochemistry tests to identify enteric bacteria,including enteropathogenic Escherichia coli (EPEC),enterotoxigenic Escherichia coli (ETEC),enteroinvasive Escherichia coli (EIEC) and enterohemorrhagic Escherichia coli (EHEC).Kirby-Bauer method was used to identify the antibiotic sensitivity.Difference of means between groups was compared by chi-square test.Results Of 2 071 enrolled children,DEC were identified in 145 (7.0 ％)cases.148 strains were isolated with three of mix infection strains.All DEC isolates in this study included 106 (71.6％) EPEC,24 (16.2％) ETEC,16(10.8％) EIEC and 2(1.4％) EHEC.The median ages of diarrheal children with DEC infections were 14 months (range:3 months to 13 years) and 62.8％ of them were ＜2 years.Among 125 DEC isolates tested for antimicrobial susceptibility,the rates of resistance to ampicillin,trimethoprim-sulfamethoxazole,cefotaxime,cefepime,gentamicin,ceftazidime,amoxicillinclavulanate,ciprofloxacin,and ofloxacin in a descending order were 55.2％,35.2％,28.0％,27.2％,23.2％,8.8％,5.6％,4.0％ and 4.0％,respectively.Resistance rates of EIEC to cefotaxime,cefepime and ceftazidime were 50.0％,43.8％ and 25.0％,respectively,which were higher than those of EPEC,ETEC and EHEC.Conclusion DEC is the important enteric bacteria that causes bacterial diarrhea in children in this study.
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<p><b>OBJECTIVE</b>To study the epidemiological characteristics and antibiotic resistance of Salmonella enterica serovar Pomona (S. Pomona).</p><p><b>METHODS</b>Antimicrobial susceptible testing (AST) and pulsed field gel electrophoresis (PFGE) methods were used to analyze on S. Pomona strains that were isolated from diarrhea cases through the diarrhea network monitoring program, environment and food samples in Shanghai as well as from reptiles in Guangxi Zhuang Autonomous Region.</p><p><b>RESULTS</b>4 553 clinic Salmonella (S.) strains were isolated from the Shanghai network laboratories from 2005 to 2012. The top 10 serotypes would include 20 serotypes all belonged to A-F groups, while S. Pomona was next to S. Wandsworth, according to the non- A-F groups. Young children seemed to be susceptible to S. Pomona, and might cause bloody stools and super-infection. The top 10 serotypes from 1 805 foodborne Salmonella strains were significantly more extensive than those from the human S. Pomona strains, followed by those rare serotypes which were mostly isolated from turtle, sea-shellfish and reptiles. Antibiotic resistance of S. Pomona strains from other sources were significantly more severe than those from human samples, and belonged to A and B clones by means of PFGE. Clone A strains were non-epidemic strains which showed multi-drug resistance (MDR) to antimicrobials. Clone B was the main epidemic-causing strain that not resistant to drugs, which consisting B- I from young-age-groups and B-II were from the seniors. B-I strains were homologous to those from shellfish, tortoises and lizards, while B-II strains only showing homology to those from shellfish. One S. Pomona strain-MDR, isolated from human was homologous to 8 antimicrobials.</p><p><b>CONCLUSION</b>S. Pomona was a quite common serotype among those rare serotypes, which showed higher pathogenicity to infants while genetic evolution might take place when comparing them with the strains isolated from the clinics in 2005. Surveillance programs should be intensified along with the early warnings systems on infections which were from seafood and reptiles.</p>
Subject(s)
Humans , China , Epidemiology , Drug Resistance, Multiple, Bacterial , Molecular Epidemiology , Salmonella Infections , Epidemiology , Microbiology , Salmonella enterica , Classification , SerogroupABSTRACT
10.3969/j.issn.1000-3606.2013.06.018
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AIM: To study the effect and the mechanism of crenulatin, an effective constituent of Chinese traditional medicine, on apoptosis of cerebral microvascular endothelial cells. METHODS: The following terminal concentrations of crenulatin were used in the study: 25 mg/L and 100 mg/L. Apoptosis of mouse cerebral microvascular endothelial cells (bEnd. 3 cell line) was evaluated by flow cytometer, immunocytochemical assay (Fas, Bcl - 2) and Western blotting (caspase - 3) after culture for 24 h. RESULTS: Compared with control group, apoptosis of bEnd. 3 cells in 25 mg/L group was significantly inhibited ( P ＜0.05), but apoptosis in the 100 mg/L group was significantly increased (P ＜ 0.05). In apoptosis inhibited group, the Fas immunocytochemical staining was weaker, the positive cells were significantly decreased ( P ＜ 0.05) and caspase - 3 expression was decreased compared with control group; however, the Bcl - 2 staining was stronger and the positive cells were significantly increased ( P ＜ 0.05). On the other hand, in apoptosis increased group ( 100 mg/L group), the changes were just opposite. CONCLUSIONS: The effect of crenulatin on apoptosis of mouse cerebral microvascular endothelial cells possesses a dual - direction change, inhibitive effect in 25 mg/L and stimulative effect in 100 mg/L group, respectively. The mechanism is related to the alterations of Fas/Bcl - 2 expression and caspase - 3 activity.
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AIM: To study the apoptotic effect of TNFα on rat pulmonary microvascular endothelial cells (PMVEC) and the influences of Fas, NF-κB in its mechanism. METHODS: Apoptosis of PMVEC was analyzed and quantitated with TUNEL, flow cytometer. The distribution of NF-κB was detected via histoimmunochemical staining in TNF-treated cells and the control. Northern blot was applied to assess the influence of TNF on PMVEC Fas expression. Fas antibody was used to investigate the apoptotic effect of Fas on PMVEC. Activation of caspase-8 was detected with Western blot. Expression of caspase-3 was analyzed with histoimmunochemical staining. RESULTS: After treatment with 5×108 U/L TNF for 24 hours, viable PMVEC significantly diminished. Apoptosis rate was 14.0%±3.1% detected with TUNEL, and 13.1% with flow cytometer. Histoimmunochemical staining showed that NF-κB relocated from cytoplasm to the nuclear. When the cells were co-cultured with TNF and APDC, an NF-κB inhibitor, less cells were viable and more cells were positively stained with TUNEL. Fas expression in PMVEC was elevated treated with TNF. Apoptosis in PMVEC was found aggravated, when the cells were co-cultured with TNF and anti-Fas antibody. The positive rate was 24.1%±1.5% with TUNEL. Increase of caspase-8 activation was manifested by Western blot following TNF stimulation. Caspase-3 expression was found elevated using histoimmunochemical staining. Cell permeable caspase-3 inhibitor significantly ameliorated PMVEC apoptosis induced by TNF. CONCLUSION: 1. Large dose of TNF(5×108 U/L) can induce apoptosis in rat PMVEC. 2. NF-κB has a protective effect on PMVEC apoptosis. 3. TNF up-regulates Fas expression in PMVEC. And the latter takes a part in apoptosis. 4. TNF induced caspase-8 activation in PMVEC, and more caspase-3 was expressed. These may be involved in PMVEC apoptosis induced by TNF.
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Microvascular endothelial cell (MVEC) is one of the target cells of TNFα (TNF effect). The dysfunction of MVEC induced by TNF plays an important role in some cardio-cerebral vascular diseases. ① Cell proliferation kinetic: Using flow cytometry, we found cell count ［(4.30±0.34)×107／L)］ in TNF group (4×105 U/L) was obviously less than that in control ［(5.23±0.50)×107／L, P＜0.01］. The cells of G1 phase were more than those of the control, while the cells of G2, S and M phase became less (P＜0.05). ② Coagulant and anticoagulant: 72 h after MVEC cultued in the media, the content of 6-keto-PGF1α (RIA) and activity of PAI decreased significantly in TNF (4×105 U/L) group (P＜0.01, vs control). The difference between TXB2 content and t-PA activity in groups was not significant (P＞0.05). ③ Adhesive molecule: The effect of low concentration TNF (＜4×105 U/L) on adhesion between cultured MVEC and leukocytes was not signficant, but when the concentration of TNF reached 8×105 U/L or more, 12 h after culture the adhesion rate between MVEC and neutrophil increased 30.8%±4.5%. If adding monoclonal antibody of ICAM-1/CD11 into media, the adhesion rate of leukocytes decreased significantly (from 31.2% to 63.4%). ④ NO: The level of nitrite in culture media (Griess reaction) was higher than that of control (P＜0.05) after pretreatment of TNF (2×106 U/L) for 6 h. Adding L-NMMA, Dexamethasone or Cycloheximide in media could block the increase of nitrite induced by TNF, while L-Arg could enhance it. The expression of iNOS mRNA of PMVEC increased significantly after treated with TNF (2×106 U/L) for 24 h (quantitative RT/PCR). Pretreatment with Dexamethasone or Cycloheximide could block the increase (P＜0.05). Meanwhile, the expression of eNOS mRNA decreased significantly compared with control, the decrease can be blocked by Cycloheximide but not by Dexamethasone. So that TNF can induce the expression of iNOS mRNA in PMVEC, but inhibited the expression of eNOS mRNA. NO production in PMVEC can be time-dependently induced by TNF. ⑤ Apoptosis: Adding high concentration TNF(＞1.2×106 U/L) in culture media for 12 h can induce apoptosis of PMVEC with electron microscopy, flow cytometry (PI/AnnexinV stain), TUNEL and DNA ladder eletrophoresis. Meanwhile, expression of apoptosis-associated gene Bcl-2 mRNA decreased and that of Fas mRNA increased (Northern blot). The expression of FADD, caspase 3 and caspase 8 enhanced too. So that signal of apoptosis induced by TNF may be transmitted following the cascade of Fas→FADD→caspase8→caspase3. In conclusion, it should be paid attention to metabolism inability and dysfunction induced by TNF which can not found easyily using light microscope. High concentration TNF can induce apoptosis of endothelial cells regulated by apoptosis-associated genes. The changes mentioned above are common pathway in pathogenesis of some diseases related to TNF.
ABSTRACT
AIM：To investigate the role of NF-κB in the activation of inducible nitric oxide synthase (iNOS) gene by tumor necrosis factor alpha (TNF α) and lipopolysaccharide (LPS) in endothelial cells and effect of antioxidant on the induction of iNOS. METHODS：Rat pulmonary microvascular endothelial cell (RPMEC) was cultured and the cells were identified with antiendothelial cell antibody CD31 using immunohistochemistry(ABC). The concentration of nitrite in the culture media was determined based on Griess reaction. iNOS mRNA was analyzed using RT-PCR and Northern blot. NF-κB in cell nuclei was detected with electrophoresis mobility shift assay (EMSA). RESULTS：A marked production of nitrite in RPMECs was found after 24 hours treatment with TNF α(105 U/L) and LPS (1 mg/L) (P＜0.01). The level of iNOS mRNA increased significantly after adding TNF α(105 U/L) and LPS (1 mg/L) to the cell media for 2 hours (P＜0.05). Pretreatment with cycloheximide (CHX, 10 mg/L) or antioxidant, PDTC (0.1 mmol/L) or NAC (20 mmol/L) significantly decreased nitrite production and iNOS mRNA expression induced by TNF α(105 U/L) and LPS (1 mg/L) (P＜0.05). Furthermore, there was a dose-effect relationship between PDTC/NAC and inhibitory effect. TNF α (105 U/L) and LPS (1 mg/L) triggered the activation and translocation of NF-κB. This effect was blocked by adding PDTC (0.1 mmol/L) or NAC (20 mmol/L) to the cell media for 1.5 hours.CONCLUSION：1.TNFα and LPS may induce iNOS gene expression at transcriptional or posttranscriptional level. The upregulation of iNOS depends on new protein synthesis. 2. The induction of iNOS gene expression by TNFα and LPS is dependent on the activation of NF-κB. 3. Antioxidants may inhibit the induction of iNOS gene through the inhibition of NF-κB activation.
ABSTRACT
AIM: To study the effects of VEGF over-expressed C6 glioma cells on the expression of Flk-1 and Flt-1 in cocultured microvascular endothelial cells using the rat hepatic cells BRL 3A as control. METHODS: Cocultured systems of rat pulmonary microvascular endothelial cells with C6 and endothelial cells with BRL 3A were established. Immunocytochemical method was used to investigate the expression change of Flk-1 and Flt-1 protein in cocultured microvascular endothelial cells. The expression of Flt-1 and Flk-1 mRNA was analyzed by RT-PCR and Northern blot. RESULTS: The microvascular endothelial cells cocultured with C6 showed increased expression of Flk-1 and Flt-1 protein ( P