Evaluation the effect of MALDI-TOF MS on the detection of carbapenemase produced by enterobacteriaceae / 中华检验医学杂志

Objective To evaluate the effect of matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS)on the detection of carbapenemase producing enterobacteriaceae. Methods A total of 21 carbapenem non-susceptible enterobacteriaceaeclinical strains were collectedfrom the Second Affiliated Hospital of Soochow University during January to May, 2018, including 11 strains of Klebsiella pneumonia, 3 strains of Klebsiella oxytoca, 3 strains of Enterobacter cloacae, and 4 strains of Escherichia coli. All the isolates were incubated with 0.5g/L meropenem solution for 2 hours. The supernatant was centrifuged and collected for MALDI-TOF MS detection. The characteristic peaks were captured to determin whether the strain was producing carbapenemase or not. And then, the results were compared with PCR results by Kappastatistical analysis. Results The PCR results showed that all the strains were positive for carbapenmase genes, among them 15 isolates were encoding KPC genes, 6 isolates encoding GES genes, 2 isolates encoding NDM genes, 1 isolate encoding VIM genes, 4 isolates encoding GIM and 1 isolate encoding SIM. And the strains could carry one or more carbapem-resistant determinants. MALDI-TOF MS showed that meropenem were hydrolyzed by 21 isolates and a characteristic drug hydrolysis peak appeared at 199 m/z, as a result of carbapenemase produced by enterobacteriaceae. The assay of MALDI-TOF MS was highly consistentwith PCR results. Conclusions The investigation showed that MALDI-TOF MS can directly detect the carbapenemase by capture the characteristic drug hydrolysis peak.

3-Methyladenine ameliorates CCl4-induced liver fibrosis and gut microbiota in mice / 临床检验杂志

Objective@#To investigate the role of gut microbiota in the amelioration of liver fibrosis induced by CCl4 in mice by 3-methyladenine (3-MA). @*Methods@#Fifteen mice were randomly divided into normal control group, liver fibrosis group and 3-MA treatment group. The liver fibrosis model was established by injecting CCl4, and the mice in the 3-MA treatment group were injected 3-MA additionally from the third week onwards. After 8 weeks, all of the mice were sacrificed and their blood, liver tissue and fecal samples were collected to analyze serum ALT, AST, GGT levels, liver histopathology and gut microbiota. @*Results@#Compared with the liver fibrosis group, serum ALT and AST levels in 3-MA treatment group decreased obviously ([68.6±4.2] U/L vs [111.0±7.8] U/L, [179.0±12.9] U/L vs [253.2±26.7] U/L, P＜0.01), and the degree of hepatic histopathological changes was reduced. The intestinal flora in three groups were distinguished by principal coordinate analysis (PCoA) and non-metric multi-dimensional scaling (NMDS) analysis. Compared with the normal control group, there were decreased Alpha diversity of intestinal community, reduced significantly abundance of Lachnospiraceae, and increased obviously abundance of Actinobacteria and Desulfovibrionacea in the liver fibrosis group (P＜0.05). Compared with the liver fibrosis group, there were increased Alpha diversity of intestinal community, increased significantly abundance of Lachnospiraceae and Blautia, and reduced abundance of Bifidobacteriaceae in the 3-MA treatment group (P＜0.05). In addition, the abundance of Lactobacillaceae in the 3-MA treatment group was significantly higher than that in the normal control group (P＜0.05). @*Conclusion@#3-MA improves the liver fibrosis of mice induced by CCl4, and gut microbiota may play an active role in this process.

The significance of glomerular fibrinogen deposition in childhood Henoch-Sch?nlein purpura nephritis / 临床儿科杂志

Objective To explore the significance of glomerular fibrinogen (Fib) deposition in Henoch-Sch?nlein purpura nephritis (HSPN). Methods The clinical and pathological data of 82 children with HSPN diagnosed by renal biopsy were retrospectively analyzed. The clinical and pathological characteristics of each group were compared according to whether there were glomerular Fib deposition and deposition intensity in the kidney. Results Glomerular Fib deposition was observed in 63 cases (76.83%) in 82 cases, including Fib+23 cases (28.05%), Fib++37 cases (45.12%) and Fib+++3 cases (3.66%), and no deposition had been found in renal tubules. The levels of high sensitivity C reactive protein (hs-CRP), D-Dimer (DD), CD19+CD23+lymphocyte subsets, and urinary albumin to creatinine ratio (UMA/Cr) were significantly different among non-deposition group, mild deposition group and severe deposition group (P<0.01). The levels of hs-CRP and CD19+CD23+in severe deposition groups were significantly higher than those in the mild and non-deposition groups (P<0.05). The level of D-D in the severe and mild deposition group was significantly higher than that in the non-deposition group (P<0.05). The UMA/Cr in the severe deposition group was significantly higher than that in the non-deposition group (P<0.05). Glomerular Fib deposition is positively correlated with IgA deposition (r=0.64, P<0.001). The proportion of glomerular IgG deposition among three groups was significantly different (P<0.05), and the proportion of IgG deposition in the severe group was the highest. Conclusions When children had glomerular Fib deposition, especially in the case of severe Fib deposition and immune dysfunction, inflammatory response and hypercoagulability are more serious and renal function damage may be more serious.

Clinical analysis of 81 children with urinary system injury after hematopoietic stem cell transplantation / 临床儿科杂志

Objectives To investigate the etiology, renal pathology, treatment, and prognosis of children's urinary system injury after hematopoietic stem cell transplantation (HSCT). Methods Clinical data of 81 children with urinary dysfunction after HSCT admitted to the Hematology Department in Children's Hospital of Soochow University were analyzed, and relevant literatures were reviewed. Results In 81 cases (50 males and 31 females), the age ranges from 8 months to 17 years old. Thirty cases (37%) with prerenal injury were recovered after active rehydration and other symptom specific treatment. There were 9 (11.1%) children with renal injury, four cases were given up therapy or transferred to other hospitals, thus lead to an unknown prognosis. Kidney biopsy was performed in the remaining five cases for pathological investigation. After active symptom-speific and etiology-based treatment, serum creatinine and glomerular filtration rate of four cases return to normal. But in the long-term follow-up,one case died of recurrence of primary disease, reinfusion of hematopoietic stem cell combined with renal failure. The remaining 3 patients were with chronic kidney disease (CKD). One case with renal thrombotic microangiopathy was in the chronic dialysis. Postrenal renal injuries were mainly hemorrhagic cystitis (28.4%) and urinary tract infection (16%). After a large dose of rehydration, urine alkalization and anti-infection therapy, they were recovered in the short term with a good prognosis. Conclusions Urinary injury after HSCT is mainly divided into three categories: prerenal, renal and postrenal, in which renal injury is prone to frequent recurrence.

Analysis of the hemolysin and virulence-related genes of incomplete hemolysis Staphylococcus aureus / 中华临床感染病杂志

Objective To analyze hemolysin and virulence -related genes in incomplete hemolytic Staphylococcus aureus.Methods Fifty strains of incomplete hemolytic Staphylococcus aureus were isolated from patients admitted in the Second Affiliated Hospital of Soochow University during 2013 and 2014, and the isolates with complete hemolytic phenotype were also collected at the same period as the control strains . All the strains were inoculated and subcultured on four kinds of sheep blood agar plates supplied by different manufacturers to compare their hemolytic phenotype .The relative mRNA expressions of hemolysin genes (hla, hlb, hlc, hld) in standard strain, complete and incomplete hemolytic phenotype strains were detected by real-time quantitative polymerase chain reaction (RT-qPCR), and valued by 2 -△△Ct method.t test was used to compare mRNA expressions of hemolysin genes .Western blot was performed to analyze the expression of α-hemolysin.Antibiotic susceptibility test of incomplete hemolytic strains was performed using broth microdilution method.Resistant gene mecA and virulence genes pvl, tst were detected by PCR.Results The steady and hereditary incomplete hemolysis was observed in 50 strains of incomplete hemolytic Staphylococcus aureus on the sheep blood agar plates from different suppliers .Taking mRNA expression of hla, hlb, hlc, hld in standard strain as 1, the relative mRNA expressions of hemolysin genes in incomplete hemolytic strains were 0.02, 7.51, 0.06 and 0.12 respectively, there were statistical differences between standard strain and incomplete hemolytic strains (t =8.46, -56.40, 8.12 and 7.61, all P <0.05).And the expression of α-hemolysin was decreased in incomplete hemolytic strains .All the strains were identified as methicillin resistant Staphylococcus aureus (MRSA).Three strains exhibited different minimum inhibitory concentrations of teicoplanin and linezolid after subcultured , but the differences had no impact on the final results of antibiotic susceptibility test .mecA, pvl and tst genes were positive in incomplete hemolytic strains . Conclusion Staphylococcus aureus with incomplete hemolytic phenotype is methicillin resistant with higher expression of β-hemolysin and lower expressions of α-hemolysin, γ-hemolysin and δ-hemolysin.It carries plv and tst virulence genes and is of high virulence .

Urinary protein markers predict the severity of renal histological lesions in children with IgA nephropathy and IgM nephropathy / 中华实用儿科临床杂志

Objective To compare the clinical manifestations,renal histological lesions,and the levels of urinary protein markers between the children with IgA nephropathy (IgAN) and those with IgM nephropathy (IgMN), and to determine whether urinary protein markers could predict the severity of renal histological lesions in children with IgAN and IgMN.Methods Seventy-four children with renal biopsy-proven IgAN and IgMN from January 2002 to October 2014 were enrolled in the study.The levels of IgG, albumin (Alb), transferrin (TRF), α1-microglobulin (α1-MG) ,β2-microglobulin (β2-MG) and N-acetyl-β-glucosaminidase (NAG) in morning urine samples before biopsy were measured.The semi-quantitative scores of mesangial hypercellularity (MC), glomerulosclerosis (GS), and tubule-interstitial damage (TID) were used to assess renal histological lesions.Multivariate Logistic regression analysis was used to determine whether urinary protein levels were independently associated with renal histological lesions.The area under the receiver-operating-characteristic curve (AUC) was calculated to assess the predictive ability of urinary protein markers.Results Seventy-four children (44 cases with IgAN,30 cases with IgMN) were included.The urinary levels of α1-MG and Alb were significantly higher in children with IgAN as compared to those with IgMN.The differences, however, did not remain significant after adjustment for age.The urine protein, as an independent factor associated with severe MC(＞ 5 mesangial cells per mesangial area) was TRF(B =0.010), and severe GS (≥ 10％ glomeruli showing segmental adhesion or sclerosis) was significantly correlated with Alb(B =0.001) ,and severe TID (focal or diffuse tubular and interstitial lesions) was significantly correlated with NAG(B =0.038).Urinary β2-MG was not significantly associated with severe MC, GS and TID.Urinary TRF, Alb and NAG achieved the best AUC of 0.85 (P ＜ 0.001) ,0.78 (P =0.002), and 0.78 (P =0.003), respectively, for predicting severe MC, GS, and TID.Conclusions Urinary proteins are useful to predict the severity of renal histological lesions in children with IgAN and IgMN.Urinary TRF, Alb and NAG have better predictive value.

Factors analysis of postoperative complications of elderly patients with gastric cancer / 中国综合临床

Objective To investigate the factors of postoperative complications of elderly patients with gastric cancer. Methods One hundred and fifth elderly patients with gastric cancer were selected as our subjects,who were underwent surgical treatments in general surgery department Jiangsu Shengze Hospital from Jan. 2009 to Dec. 2012. The general informations including age,staging of gastric cancer,radical operation performed or not,resection combined with other organs and operation periods were recorded. Results There were no significant differences between postoperative complications patients in terms of age,staging of gastric cancer,radical operation performed or not,resection combined with other organs( χ2 = 3. 113,P = 0. 078,χ2= 0. 465,P = 0. 495,χ2 = 0. 518,P = 0. 472,χ2 = 0. 303,P = 0. 582). The cases with operation periods 3 h were 22(42. 3% )and 12(19. 1% )respectively,and the difference was significant( χ2 = 6. 326,P= 0. 012). Conclusion Operation period is the main influencing factor of the elderly patients with gastric cancer. Therefore,reducing operation period under ensuring operation quality is more efficiency to decrease the complication.

Biological characteristics of rpoS gene deleted mutant in Salmonella typhi / 中华临床感染病杂志

Objective To investigate the biological characteristics of rpoS gene deleted mutation in Salmonella typhi under different stress conditions,so as to explore the target gene for the prevention and treament of Salmonella typhi infection.Methods rpoS gene deleted mutant of Salmonella typhi was prepared by homologious recombination.rpoS mutant and parental strains were incubated under iso-osmia and various stress conditions:acid stress(pH 4.2),high osmolarity stress(NaCl 300 mmol/L),bile stress (1.5 mmol/L sodiumdeoxycbolate)and oxidative stress(1 mmol/L H2O2).The growth curves were compared between mutant and parental strains under different incubation conditions(t test).Results rpoS gene deleted mutant of Salmonella typhi Was successfully generated.Compared with the parental strain,the survival ability of rpoS mutant was significantly compromised under the acid stress,high osmolarity stress and oxidative stress(t values at4 h were 12.864,3.594 and 12.979;t values at 14 h were6.497,3.039 and 10.440,P<0.05 or<0.01).Conclusion rpoS is important for Salmonella typhi to overcome the acid,high osmolarity and oxidative stresses,and it may be a target gene for the prevention and treatment of Salmonella typhi infection.

Regulative role of specific cytokine on expression of T-helper 17 cells and regulatory T cells / 中华检验医学杂志

Objective To investigate the roles of a variety of cytokines including transforming growth factor beta (TGF-β),interleukin-6 (IL-6) and interleukin-2 (IL-2) in the differentiation of CD+4 Tlymphocyte cells.Methods T lymphocyte cells either in human peripheral blood or routine spleen were cultured in vitro under different stimulation conditions.Flow cytometry was used to analyze the percentages of CD+4IL-17+ T-helper 17(Th17) cells,CD+8 IL-17+ T cells,CD+4 CD+25 FOXP+3 T regulatory (Treg) cells among activated T cells.Results Differentiation of Treg cells,Th17 cells and CD+8 IL-17+ T lymphocyte cells was enhanced when murine splenic T cells were cultured with TGF-β.The levels of expression were (7.8±2.2)%,(12.6±3.1)%,(10.1±2.6)% ,respectively.Experimental control group was severally same type of T cells without cytokine treatment.The levels of expression were (4.8±0.6) %,(1.7±0.5) %,(1.0±0.4) %,respectively.There were statistically significant differences among them (q=4.09,8.80,9.61.P<0.05 or P<0.01).Under combination treatment with IL-6 and TGF-β,(17.8±5.3) % Th17 cells and (15.0±4.2)% CDCD+8 IL-17CD+ T cells were induced,whereas the levels of Treg cells whose differentiation were restrained were (4.1±1.2) %.The differences were statistically significant compared with the level of same type of T cells in TGF-β group (q=5.03,5.17,5.04,P<0.01).Moreover,combination treatment with IL-2 and TGF-β decreased percentages of Th17 and CDCD+8 IL-17CD+ T cells and increased percentages of Treg cells in T cell population.There was an opposite effect when anti-IL-2 was apphed.The percentages of Th17 and CD+8 IL-17+ T cells were increased and the percentages of Treg cells were reduced The regulation trend of T lymphocyte cells in human peripheral blood was similar with those in routine spleen.Conclusion Various cytokines are of great importance in the regulation of the balance between Th17 and Treg cell.

Drug Resistant Genes in ICU Isolates of Pseudomonas aeruginosa and Cluster Analysis of Strains / 中华医院感染学杂志

OBJECTIVE To investigate the existence of genes for beta-lactam antibiotic resistance and for aminoglycosides modification enzymes(AMEs) in Pseudomonas aeruginosa(PAE) isolates from ICU patients and analyze the homology among strains.METHODS ?-Lactamase genes including TEM,SHV,OXA-10,PER,VEB,GES,CARB,IMP,VIM,SPM,GIM,DHA,FOX,MOX and oprD2,were detected by PCR amplication in 21 PAE isolates.The genes for AMES including aac(3)-Ⅰ,aac(3)-Ⅱ,aac(6′)-Ⅰ,aac(6′)-Ⅱ,ant(3″)-Ⅰ and ant(2″)-Ⅰwere determined by PCR amplification as well.RESULTS Among 21 isolates 21(100%),2(9.5%),1(4.8%),2(9.5%)and 4(19.0%) were positive for TEM,SHV,GES,CARB and VIM genes,respectively.The deletion of oprD2 gene was found in 14 out of 21 strains.Other ?-lactamase genes were absent in all isolates.As for AME genes,aac(3)-Ⅱ,aac(6″)-Ⅰ,aac(6)-Ⅱ,ant(3″)-Ⅰ,ant(2″)-Ⅰ and aac(3)-Ⅰgenes were present in 19.0%,23.8%,9.5%,4.8%,and 19.0% of 21 isolates,However,aac(3)-Ⅰ gene was no position in any isolates.CONCLUSIONS P.aeruginosa carries various beta-lactamase and AME genes in ICU patients.Genetic cluster analysis suggested that clonal propagation result in nosocomial infection of PAE.

Clinical significance of platelet-leukocyte aggregates in patients with acute cerebral infarction / 临床检验杂志

Objective To research the clinical significance of leukocyte-platelet aggregates(PLA)in patients with acute cerebral infarction.Methods Sixty-five patients with acute cerebral infarction and 24 controls.The levels of PLA,platelet-neutrophil aggregates(PNA),platelet-lymphocyte aggregates(PLyA)and platelet-monocyte aggregate(PMA)were analyzed by two-color flow cytometry,soluble P-selectin(sP-sel)was detected by ELISA,serum C-reaction protein(CRP)was detected by nephelometry and platelet aggregation rate(PAR)was measured by blood agglutination.Results There were no differences in the percentages of PLA,PNA and PLyA between patients(4.12?1.17%,4.09?1.27%,4.28?1.09%)and controls(4.10?0.99%,3.98?1.12%,4.01?1.98%).However,in the patients before treatment,the percentage of PMA(11.54?1.88%)was significantly increased compared with that after treatment(8.40?2.74%,P

Incidence and Drug-Resistance of Helicobacter pylori from Patients with Peptic Ulcer / 中华医院感染学杂志

OBJECTIVE To investigate incidence and drug susceptibility of Helicobacter pylori(Hp) in local area.METHODS Strains of Hp were isolated and cultivated from gastric antrum mucosa of 141 peptic ulcer disease(PUD) patients diagnosed by gastroscope.The drug sensitive test of Hp strains was examined by K-B method.RESULTS The isolation rate of Hp in 141 PUD patients was 75.2%.Hp isolates showed a relative hypersensitivity to clinical commonly used drugs(ie furazolidone,amoxicillin,levofloxacin,cefuroxime and moxifloxacin).On the other hand,Hp isolates exhibited resistance to metronidazole and clarithromycin.CONCLUSIONS Hp is an important etiologic factor of PUD,and showing an increasing resistance to common clinical antibiotics.

Genes for ?-Lactam Antibiotic Resistance in Clinical Isolates of Pseudomonas aeruginosa / 中华医院感染学杂志

OBJECTIVE To investigate genes associated with the drug-resistance of ?-lactam antibiotics in Pseudomonas aeruginosa(PAE) isolated from clinical patients. METHODS ?-Lactamase genes including TEM,SHV,OXA-10,PER,VEB,GES,CARB,IMP,VIM,SPM,GIM,DHA,FOX,MOX and oprD2 were detected by PCR amplification in 33 PAE isolates. RESULTS TEM,SHV,GES,CARB and VIM genes were positive in 100%,6.1%,6.1%,9.1% and 12.1% of 33 isolates,respectively.The deletion of oprD2 gene was found in 22 isolates.Other ?-lactamase genes were absent in all isolates.By PCR amplification,DNA sequencing and BLASTn comparison analysis,the CARB genes of 2 strains were demonstrated to be CARB-3 and the VIM genes of 2 strains were VIM-2. CONCLUSIONS P.aeruginosa carries various beta-lactamase genes in clinical PAE patients,and the deletion ratio of oprD2 gene is high.

Aminoglycosides Modification Enzymes Genes in Pseudomonas aeruginosa Isolated from ICU Patients / 中华医院感染学杂志

OBJECTIVE To investigate genes associated with aminoglycosides modification enzymes(AMEs) in Pseudomonas aeruginosa(PAE) isolated from ICU patients.METHODS Drug-reisistant genes encoding AMEs such as aac(3)-Ⅰ,aac(3)-Ⅱ,aac(6′)-Ⅰ,aac(6′)-Ⅱ,ant(3″)-Ⅰ and ant(2″)-Ⅰ were detected by polymerase chain reaction(PCR)(amplification) in 21 PAE isolates.RESULTS The positive rates of aac(3)-Ⅱ,aac(6′)-Ⅰ,aac(6′)-Ⅱ,ant(3″)-Ⅰ,ant(2″)-Ⅰ and aac(3)-Ⅰ genes were positive in 19.0%,23.8%,9.5%,4.8%,19.0% and 0% of 21 isolates,respectively. Drug-resistant genes encoding AMEs were detected positively in 42.8% of 21 isolates.(CONCLUSIONS) AMEs genes are present in high percentage of PAE isolated from ICU patients.