Surveillance of molecular characteristics of gyrA and parC among fluoroquinolone resistant Streptococcus agalatiae isolates in Suzhou hospitals / 国际检验医学杂志

Objective To investigate the molecular characteristic of fluoroquinolone resistant Streptococcus agalactiae (GBS) isolates in Suzhou.Methods Totally 46 fluoroquinolone resistant GBS strains were collected,and then subjected into PCR and two resistance genes were sequenced and their high frequency mutation sites,gyrA and parC were analysed.Results Among these isolates,the most frequent gyrA mutation was gyrA_S81L (TCA→TTA),which displayed 93.4％ (43/46) with substituted at No.81 site of amino acid from Ser to Leu.However,the most frequent parC mutation was mutated at No.79 site of amino acid from Ser to Tyr orPhe,with two mutations parC_S79Y (73.9％,34/46) and parC_S79F(17.4％,8/46).The analysis showed that the major mutation patterns are gyrA_S81L with parC_S79Y (73.9％,34/46),gyrA_S81L with parC(13.0％,6/46).Conclusion The most frequent mutations of fluoroquinolone resistant gene in Suzhou are gyrA and parC and the major mutation patterns are gyrA_S81L/parC_S79Y and gyrA_S81L/parC_S79F.

The role of glutamate and peptidoglycan in nanotubes formation between adjacent bacteria / 中国人兽共患病学报

We investigated the formation mechanism and immediate constituents of nanotubes between adjacent bacteria,pre venting rapid increase of drug-resistant bacteria provide potential targets.We used scanning electron microscopy(SEM) to observe formation of nanotubes;glutamic was determined by surface desorption atmospheric pressure chemical ionization mass spectrometry(SDAPCI-MS)and peptidoglycan was determined by enzyme linked immunosorbent assay(ELISA) in the forma tion of nanotubes before and after.The results showed that Staphylococcus aureus and Escherichia coli consumed more glutamic after formation of nanotubes;at the same time,the concentration of peptidoglycan in nanotubes formation of S.aureus and E.coli increased significantly.This study illuminated the glutamic and peptidoglycanin role mechanism of nanotubes form.We found that peptidoglycan is constituents of nanotubes and glutamic is the main energy source for the formation of nanotubes.

Predictive effect of neonatal morbidities on the poor outcomes at 12 months corrected age in very low birth weight premature infants / 中华儿科杂志

Objective@#To investigate the prognostic effect of neonatal morbidities on poor outcomes at 12 months corrected age in very low birth weight (VLBW) premature infants .@*Method@#From November 2013 to October 2014, a multi-center retrospective study was conducted in 8 tertiary Maternal and Children′s hospitals in Guangdong, Hunan and Fujian. The premature infants survived to a postmenstrual age (PMA) of 36 weeks with birth weight less than 1 500 g and without congenital diseases were included, and divided into two groups according to poor outcomes. The birth weight, gestational age, morbidities and poor outcomes (death, cerebral palsy, cognitive delay, et al) were recorded. Data were analyzed with Chi-square test to investigate the relationship between morbidities and poor outcomes. And the predictive effect of the top three morbidities were analyzed by Logistic regression analysis.@*Result@#Total of 834 VLBW premature infants (473 boys and 361 girls) finished the follow-up, whose average gestational age and birth weight were (30.6±1.8) weeks and (1 189±159)g. The incidences of BPD, severe ROP, NEC, brain injury and sepsis were 207 (24.8%), 119 (14.3%), 58 (7.0%), 281 (33.7%) and 124 (14.9%), respectively. There were significant differences between the two groups in the incidences of BPD, severe ROP, NEC, brain injury and sepsis(χ2=42.10, 47.20, 4.81, 44.28, 18.63, all P<0.01), which had significant correlation with poor outcomes at 12 months corrected age. The three top morbidities were severe ROP, BPD and brain injury(OR=3.82, 2.90, 2.80). Combined morbidities with BPD, severe ROP and brain injury correlated with higher risk of poor outcomes (one morbidity, OR=3.14, β=1.15; two morbidities, OR=7.31, β=1.99; three morbidities, OR=22.41, β=3.11; all P<0.01).@*Conclusion@#BPD, severe ROP, NEC, brain injury and sepsis were the risk factors of poor outcomes at 12 months corrected age in VLBW infants. And the more combined morbidities with severe ROP, BPD and brain injury, the higher risk of poor outcomes in this population. Trial registration Clinical Trails, NCT03104946.

Analysis of clinical infection status of carbapenem-resistant Enterobacteriaceae / 国际检验医学杂志

Objective To investigate the clinical distribution of carbapenem-resistant Enterobacteriaceae(CRE)strains separated in this hospital and the situation of its production of carbapenem enzyme.Methods The production of carbapenem enzyme by CRE strains was confirmed by using modified Hodge test,the situation of the production of metallo-beta-lactamases by CRE strains was screened by using imipenem-EDTA double-disk synergy test,and the clinical distribution of CRE strains was retrospectively ana-lysed.Results 37 strains of CRE isolated in this laboratory were screened by using instrument method and verified by using disk diffusion (K-B)method.It showed an increasing trend from 2012 to 2014 in the amount of CRE strains.In terms of bacterial spe-cies,K.pneumonia(1 6 strains)was the main kind of carbopenems-resistant strains,followed by E.coli(6 strains),Ser.marcescens(6 strains)and E.cloacae(4 strains).CRE strains were mainly isolated from geriatric ward and intensive care unit(ICU).Sputum,u-rine and blood specimen were key sources of CRE strains.Modified Hodge test confirmed that 36 strains of CRE were the strains that can produce carbapenemase,including 4 strains of K.pneumonia,3 strains of E.cloacae,and 1 strain of E.asburiae,and strains producing metallo-beta-lactamases were confirmed by using imipenem-EDTA double-disk synergy test.Conclusion Elderly patients with underlying diseases are susceptible population of CRE hospital infection and are primary preventive targets.The principal mechanism of carbapenem-resistant CRE strains in this hospital is the production of carbapenemase and production of metallo-β-lac-tamases in a small number of strains.

Pevalence rate of healthcare-associated infection in a maternal and child health care hospital / 中国感染控制杂志

Objective To understand healthcare-associated infection(HAI)in a maternal and child health care hos-pital,so as to provide scientific evidences for further targeted surveillance.Methods A cross-sectional survey was performed by bedside visiting and medical record reviewing.Results Of 768 hospitalized patients,9(1 .18%)had HAI,the top 3 highest prevalence rates were found in obstetrical intensive care unit (9.09%),neonatal intensive care unit (5.80%)and gynecological department II(2.22%).Antimicrobial usage rate was 30.34%(n=233),134 of which (57.51 %)were prophylactic use,165 were mono-therapy(70.82%).A total of 5 pathogenic bacteria were isolated,the number of Streptococcus agalactiae ,Klebsiella pneumonia ,Enterococcus faecalis ,and Staphylococcus saprophyticus was 2,1 ,1 ,and 1 respectively,except Streptococcus agalactiae ,the other 3 strains were multidrug-resistant organisms(MDROs).Conclusion Surveillance on MDRO infection should be paid much attention,the oc-currence of MDRO infection should be reduced through targeted and bundle intervention.

Evaluation of a Novel Array-Based Toxoplasma, Rubella, Cytomegalovirus, and Herpes Simplex Virus IgG Enzyme Linked Immunosorbent Assay and Its Comparison with Virion/Serion Enzyme Linked Immunosorbent Assays

BACKGROUND: The dramatic increase in use of the IgG test for toxoplasma, rubella, cytomegalovirus (CMV), and herpes simplex virus (HSV) [TORCH] has led to the requirement for a high-efficiency method that can be used in the clinical laboratory. This study aimed to compare the results of BGI-Array ELISA TORCH IgG (BGI-GBI, China) screening method to those of Virion/Serion TORCH IgG ELISA (Virion/Serion, Germany). METHODS: Serum specimens (n=400) submitted for routine IgG testing by Virion/Serion ELISA were also tested using the BGI-Array ELISA method. The agreements of these two kinds of method were analyzed by kappa-coefficients calculation. RESULTS: Following repeat testing, the BGI-Array ELISA TORCH IgG assays demonstrated agreements of 99.5% (398/400 specimens), 98% (392/400 specimens), 99% (396/400 specimens), and 99.5% (398/400 specimens), respectively. The BGI-Array ELISA IgG assays provided results comparable to Virion/Serion ELISA results, with kappa-coefficients showing near-perfect agreement for the HSV (kappa=0.87), rubella (kappa=0.92) and CMV (kappa=0.93) and substantial agreement for the toxoplasma (kappa=0.80) IgG assays. The use of the BGI-Array ELISA TORCH IgG assays could reduce the turnaround time (1.5 hr vs. 5 hr by Virion/Serion ELISA for 100 specimens) and were easy to use. CONCLUSIONS: BGI-Array ELISA TORCH IgG shows a good agreement with Virion/Serion ELISA methods and is suitable for clinical application.

Regulation effect of adipose-derived mesenchymal stem cells on pancreatic cancer cells / 中华外科杂志

<p><b>OBJECTIVE</b>To analyze the effects of adipose tissue-derived stem cells (ADSCs) on the proliferation and invasion of pancreatic cancer (PaCa) cells and the the possible mechanism involved.</p><p><b>METHODS</b>ADSCs were isolated and co-cultured with PaCa cells. CCK-8 assay was used to detect the proliferation of PaCa cells. An ELISA was used to determine the concentration of stromal cell-derived factor-1 (SDF-1) in the supernatants. The proliferation of PaCa cells by SDF-1 was measured. AMD3100 regulated the co-culture of ADSCs and PaCa. The tumor growth of PaCa cells was assessed after treatment by ADSCs in vivo.</p><p><b>RESULTS</b>ADSCs can promote the proliferation and invasion of PaCa cells (proliferation: SW1990: 1.535 ± 0.153; PANC-1: 1.370 ± 0.100; the value of control was 1; invasion: SW1990: 47.0 ± 2.6 vs. 28.3 ± 1.3; PANC-1: 40.3 ± 1.8 vs. 24.3 ± 1.3; t = 4.332-9.558, P < 0.05). The expression of SDF-1 was high in ADSCs, but not in PaCa cells (69 ± 5 vs. 0 and 0, F = 389.134, P < 0.01). The promotion of SDF-1 on PaCa cells depends on the concentration. AMD3100 significantly downregulates these growth-promoting effects of ADSCs on PaCa cells. ADSCs significantly promoted the growth of SW1990 in nude mice at the 5(th) week (volume: (1295 ± 102) mm(3) vs. (967 ± 81) mm(3), t = 5.614, P < 0.05) , but not in PANC-1 cell.</p><p><b>CONCLUSION</b>ADSCs can promote the proliferation and invasion of PaCa cells, which may involve the SDF-1/CXCR4 axis.</p>

Effects of glycogen synthase kinase 3β overexpression in rat and glycogen synthase kinase 3β inhibitor SB-216763 on proliferation of hepatic oval cells / 中华外科杂志

<p><b>OBJECTIVE</b>To research the effects of glycogen synthase kinase (GSK3β) overexpression and GSK3β inhibitor SB-216763 on the proliferation of hepatic oval cells in rats and its regulatory mechanisms by Wnt signaling pathway.</p><p><b>METHODS</b>The hepatic oval cells WBF-344 were divided into the blank control group, GSK3β over-expression group, DMSO control group and GSK3β inhibitor groups, while the inhibitor groups set up three concentration gradients, that was 1, 5, 10 µmol/L. Using the GSK3β over-expression lentivirus, which had been identified correctly, and SB-216763 dealt with the cells WBF-344. The cells morphology of each group was observed under the phase contrast inverted microscope, and the expression of fluorescence in the lentivirus-transfected group was observed under the fluorescent microscope. The proliferation of each group cells was tested by CCK8 kits. The cells' apoptosis was detected by AnnexinV-FITC/PI kits. The expression of GSK3β, β-catenin and cyclin D1 were detected by Western blot.</p><p><b>RESULTS</b>The cells of GSK3β over-expression group were fewer and obvious aging. However, in each inhibitor added group, the cells' division and proliferation was vigorous, and the condition was good. Moreover, the cells' proliferation was getting stronger with the concentration of SB-216763 increasing. A large number of green fluorescence was expressed in the lentivirus-transfected cells. The cells' proliferation in GSK3β over-expression group restrained (t = 7.178, P < 0.01, as compared with control), while the cells' proliferation was vigorous in inhibitor groups (F = 45.030, P < 0.01, as compared with control). Flow Cytometry showed that the cells apoptosis was significant in GSK3β over-expression group. Western blot showed that the expression of GSK3β was increased, while the expression of β-catenin and cyclin D1 was decreased in the over-expression group. The expression of GSK3β had no significant difference among the control group and inhibitor groups. However, the expression of β-catenin and cyclin D1 was significantly increased with the concentration of SB-216763 increasing.</p><p><b>CONCLUSIONS</b>The overexpression of GSK3β can inhibit the Wnt signaling pathway, thus restrain the cells' proliferation and promotes apoptosis. SB-216763 can activate the Wnt pathway, thus promotes cells' proliferation.</p>

Clonal Spread of Carbapenem Non-susceptible Acinetobacter baumannii in an Intensive Care Unit in a Teaching Hospital in China

BACKGROUND: This study was aimed to investigate the genetic diversity and antibiotic resistance profile of the nosocomial infection agent Acinetobacter baumannii from a medical intensive care unit (ICU) in a teaching hospital in Suzhou, China. METHODS: The genetic relationship among A. baumannii isolates in an ICU was investigated using multilocus sequence typing (MLST). The antibiotic resistance pattern was determined by performing an antibiotic susceptible test, which included an agar dilution method and an E-test method. Resistant determinants, e.g., carbapenemase genes, metallo-beta-lactamases, and class 1 integron, were analyzed by specific PCR and DNA sequencing. RESULTS: In the present study, 33 non-duplicate isolates were identified as 5 existing sequence types (STs) (ST92, ST75, ST112, ST145, and ST345) and 1 new sequence type STn, which has a G-A mutation at nt268 on ropD40 of ST251. These results reveal limited diversity in carbapenem non-susceptible A. baumannii (CNSAb) isolates in our ICU, which are comprised of only 2 distinct STs, with ST92 and ST75 clustering into a clonal complex (CC) 92. Most CNSAb isolates (94.4%, 17/18) harbored the OXA-23 gene, while no carbapenem-susceptible A. baumannii (CSAb) isolates harbored it. In addition, 66.7% (22/33) isolates were positive for class 1 integrase, and gene cassette analysis showed there are 3 gene arrays among them, i.e., aacA4-catB8-aadA1 (77.3%, 17/22), aacA4 (22.7%, 5/22), and aacC1-orfX-orfX'-aadA1 (4.5%, 1/22). CONCLUSIONS: When all these data are combined, the antibiotic resistance and wide distribution of CNSAb isolates in our ICU are probably caused by expansion of the CC92 clone.

Type IV secretion system in Helicobacter pylori: a new insight into pathogenicity / 中华医学杂志(英文版)

<p><b>OBJECTIVE</b>To review the research progress on Type IV secretion system (T4SS) in Helicobacter pylori.</p><p><b>DATA SOURCES</b>The data used in this review were identified by searching of PUBMED (1995 - 2007) online resources using the key terms 'Type IV secretion system' and 'Helicobacter pylori'.</p><p><b>STUDY SELECTION</b>Mainly original articles and critical reviews written by major pioneer investigators of this field were selected.</p><p><b>RESULTS</b>The research progress on T4SS in Helicobacter pylori was summarized. The structure and function was discussed.</p><p><b>CONCLUSIONS</b>T4SS is not only involved in toxin secretion and injection of virulence factors into eukaryotic host target cells, but also involved in horizontal DNA transfer to other bacteria and eukaryotic cells, through DNA uptake from or release into the extracellular milieu. It provides a new insight into the pathogenicity of Helicobacter pylori and a novel target for antimicrobials development. However, many challenges remain for us in understanding the biological role of T4SS in Helicobacter pylori.</p>

Cloning,Expressing and Analysis of Helicobacter pylori NCTC 11637 hp0525 Gene and Study the Influence of Proliferation of SGC-7901 Cell / 中国生物工程杂志

Objective:Helicobacter pylori is one of the human pathogens causing gastric ulcers and cancers.A key virulence factor of H.pylori is the Cag pathogenicity island,which encodes a type IV secretion system.HP0525 is an essential component of the Cag system and acts as an inner membrane associated ATPase.To construct recombinant plasmid containing hp0525 gene of Helicobacter pylori(H.pylori)NCTC 11637,and analysis of sequence nucleic acid,express it in E.coli BL21 and study the influence of the HP0525 protein on proliferation of SGC-7901 cells.Methods:H.pylor hp0525 gene was amplified from the genome DNA by PCR,then operated T-A cloning and sequenced.The hp0525gene fragment was inserted directionally into vector pMD18-T to construct recombinant clones of hp0525 and was sequenced.The recombinant plasmid was transformed into E.coli BL21for expressing under induction of IPTG.Purify the expressed protein by Ni2+-NTA column chromatography.Expressed product was analyzed by Western blot and MALDI-TOF.Added the purified protein into SGC-7901 cells,the effect of cell proliferation in SGC-7901 cells induced by the recombinant protein was observed by MTT.Results:A 993 base pairs long hp0525 gene,which encodes a product of 330 amino acid,was obtained using PCR method and was cloned into pMD18-T vector successfully.The sequence analysis for hp0525 showed that it shares 97%~99% homology with other strains in Gene bank.SDS-PAGE showed a protein band with a relative molecular weight of 36 000,which was consistent with the expectation.The expressed product reached a purity of 97% after Ni2+-NTA column chromatography.The protein after dialyzed and annealed,was co-cultured with SGC-7901 cells,The protien of different concentration co-cultured with SGC-7901 cells for different times,found that the protein in low concentration stimulates proliferation of cells,to achieve some concentration,it inhibits proliferation of cells along with multiplication of the concentration of the protein;The protein inhibits proliferation of cells relay on the extension of time and concentration.Conclusion:It is indicated that the correct hp0525 gene was obtained and expressed in E.coli BL21.High purified protein was obtained by Ni2+-NTA column chromatography,the protein can inhibits proliferation of SGC-7901cells,and T-ATPase activity which posed a basis for further researching on its biological function.

Significance of C-myc Expression in Renal Tissue in Children's Primary Nephrotic Syndrome / 中国医师杂志

Objective To observe the correlation between the C-myc expression and pathologic lesions of kidney to explore the role of C-myc re-expression in children's primary nephrotic syndrome(CPNS). Methods The C-myc expression of renal tissue in 30 cases of CPNS was detected using immunohistochemical method, and the correlation between C-myc expression and renal pathological lesions was analyzed. Results There were various degrees of C-myc positive staining in the renal tissue of all patients with CPNS, while no C-myc positive expression in the renal tissue of control group. C-myc expression was mainly located in podocytes and less in endothelial cells of glomeruli. There was high level of C-myc expression in nephric tubules, especially in proximate tubules. There was no C-myc expression in the Henle's loop,tubulous matrix and vessel areas. There was no obvious difference in C-myc expression level in the podocytes and proximate tubules among the different pathological types of CPNS. The level of focal segmental glomerulosclerosis(FSGS) in proximate tubules obviously reduced compared with mesangil proliferative glomerulonephritis(MsPGN) and minimal change disease(MCD)(P