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Objective To build predictive model of secondary mild cognitive impairment (MCI) in patients with painful diabetic neuropathy (PDN), and analyze its apply. Methods The patients with PDN were consecutively selected from March 2013 to March 2016. The relevant clinical data were recorded, and the patients were followed up for 1 year. According to the results of follow-up, secondary MCI risk indicators were predicted, and the time window of adverse outcomes event was validated. Results A total of 82 PDN patients completed the study, and secondary MCI occurred in 16 cases. Sixty-six cases had not secondary MCI. The Cox regression model multivariate analysis results showed that the independent influencing factors of secondary MCI was course of PDN, brief pain inventory (BPI) score and neuron-specific enolase (NSE) in patients with PDN (HR = 1.238, 1.336 and 1.450; P<0.05). The secondary time window of the MCI in PDN patients with the course of PDN ≥3.367 years, BPI score≥4.704 scores and NSE ≥ 7.420 μg/L was shorter, in whom BPI score and NSE had a higher evaluation ability. Conclusions The courses of PDN, BPI score and NSE are independent influencing factors of secondary MCI in PDN patients, and the BPI score≥4.704 scores and NSE≥7.420μg/L have a higher evaluation ability.
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Objective To investigate the antimicrobial resistance ,molecular phenotypes ,virulence gene profiles of Salmonella A gona (S .A gona) isolated from patients with acute diarrhea ,and to better understand its epidemic trend ,prevention and treatment .Methods Clinical data and stool samples of patients with acute diarrhea during April to October in 2013 and 2014 from the Second Hospital of Tianjin Medical University were collected .Enrichment culture ,biochemical identification and serotyping analysis were used to isolate and identify S .A gona strains .The isolated strains were further analyzed with antibiotics susceptibility test ,pulsed field gel electrophoresis (PFGE) ,multiple locus sequence typing (MLST ) , Quinolone resistance determining region (QRDR) .Plasmid-mediated quinolone resistance (PMQR) and β-lactamases genes (TEM ,SHV ,OXA ,and CTX-M) were amplified by polymerase chain reaction (PCR) and sequencing .The representative genes carried by Salmonella pathogenicity islands (SPI) 1 — 6 ,9 — 12 and virulence plasmids were amplified by PCR .And the clinical characteristics of S .Agona infection were analyzed .Results Among 119 non-repetitive (non-typhoidal salmonella ,NTS) isolates during the two years ,eight isolates (6 .7％ ) of S .A gona were identified . The resistance rate of S .A gona strains to streptomycin was 100 .0％ , those to ampicillin and gentamicin were 62 .5％ ,to levofloxacin ,ciprofloxacin and nalicixic acid were 25 .0％ ,to chloramphenicol ,amoxillin/clavulanic acid and piperacillin tazobactam were 12 .5％ .The strains were susceptible to other drugs .All 8 isolates had the identical ST13 genotype .PFGE showed 5 clones ,and 4 out of 5 isolates had the exact same patterns of PFGE and drug susceptibility .Two (fluoroquinolones ,FQ) resistant strains carried gyrA mutation leading to amino acid substitutions at position 87 in GyrA ,and no PMQR genes was detected ,while one of which was sensitive to ciprofloxacin by K-B method .All five ampicillin-resistant isolates were positive for TEM-1b gene and one isolate of them was resistant to β-lactam/β-lactamase inhibitor complex .The representative genes carried by SPI 1 — 6 , 9 ,11 ,12 (hilA ,sseL ,mgtC ,siiE ,sopB ,pagN ,bapA ,pagC and sspH2) were 100 .0％ positive ,while the genes carried by SPI10 (sef A ) virulence plasmids (spvB , prot6E) were negative . Two patients with FQ resistant strains infection were clinically diagnosed with bacillary dysentery ,and the remaining six cases with FQ susceptible strains infection were clinically diagnosed with acute gastroenteritis .Conclusions FQs-resistant and multi-drug resistant S .A gonaisolates have emerged in clinical settings .These isolates carry a variety of virulence genes .Resistance to FQ of S .Agonamay cause more severe illness .ST13 might be the dominant genotype of S . A gona in China ,and we should try to prevent the infection outbreak of S .A gona .
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Objective To investigate the clinical value of serum procalcitonin (PCT) in diagnosis of urinary tract infection(UTI) in elderly patients.Methods 114 elderly patients with UTI in the department of infectious diseases of a hospital from January 2013 to December 2014 were analyzed retrospectively, clinical data of patients with abnormal and normal serum PCT were compared, PCT levels in patients with positive and negative blood cultures were compared, PCT receiver operating characteristic (ROC) curve for the diagnosis of bacteremia were drawn.Results Among 114 elderly patients with UTI, 46 were with abnormal PCT, 68 were with normal PCT.In abnormal PCT group, the proportions of patients with highest body temperature within 24 hours of admission, white blood cell count, neutrophil granulocyte percentage, C-reactive protein (CRP), blood urea nitrogen(BUN), creatinine(Cr), and urinary tract obstructive disease were all higher than those with normal PCT (all P<0.05).Among 42 patients with blood culture, PCT level in positive blood culture group(n=12) was higher than negative blood culture group(n=30)(1.93 [0.57-8.32] μg/L vs 0.36[0.15-1.01]μg/L, P=0.028).The area under the ROC curve (AUC) of the patients with bacteremia diagnosed by PCT was 0.72(95%CI:0.54-0.90),at the optimal value of 0.52 g/L, sensitivity, specificity, positive predictive value, and negative predictive value were 83.3%, 63.3%, 47.6%, and 90.5% respectively.Conclusion Serum PCT level can well reflect the severity of elderly patients with UTI, and is of great value in early diagnosis of bacteremia in elderly patients with UTI.
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Objective To investigate the antibiotic susceptibilities and the profiles of virulence genes of clinically isolated Salmonella enterica serovars Schwarzengrund ( S. Schwarzengrund) strains for bet-ter understanding the epidemiological trend of this type of non-typhoidal Salomonella and to provide guide-lines for the prevention and treatment of S. Schwarzengrund infection. Methods Stool samples and clinical data of patients with acute diarrhea who received treatment in the Second Hospital of Tianjin Medical Univer-sity during May, 2014 to October, 2014 were collected for this study. Enrichment culture and biochemical identification were used to isolate and identify the S. Schwarzengrund strains. The isolated strains were fur-ther analyzed with serotyping analysis, drug susceptibility test, pulsed field gel electrophoresis ( PFGE) and multiple locus sequence typing ( MLST ) . The representative genes carried by Salmonella pathogenicity islands (SPI) 1-5, SPI regulators and virulence plasmids were amplified by PCR. The coding genes of CdtB-islet, which were cdtB, pltA and pltB were amplified and sequenced. Results In total, 16 (14. 8%) out of 108 non-typhoidal Salmonella strains were identified as S. Schwarzengrund strains and all of them were sus-ceptible to 11 kinds of antibiotics such as fluoroquinolone, ampicillin, ceftriaxone and trimethoprim-sulfame-thoxazole. PFGE categorized the 16 S. Schwarzengrund strains into 3 clusters including A clone ( 14 strains), B clone (1 strain) and C clone (1 strain). The strains that isolated from 8 patients who ate the same food belonged to one cluster ( A clone ) , suggesting that it was an outbreak of infection. The 16 S. Schwarzengrund strains showed identical MLST type, which was ST241. The representative genes carried by SPI1-5 ( invA, sitC, hilA, sseL, sifA, mgtC, siiE and sopB) , the regulatory gene ( phoP) and the cytole-thal distending toxin islet (CdtB-islet) coding genes (cdtB, pltA and pltB) were positive, while the genes carried by virulence plasmids (pefA, prot6E and spvB) were negative. The similarities in CdtB-islet coding genes and amino acids sequences between Salmonella typhi and S. Schwarzengrund strains in this study were more than 97% and 98%, respectively. Conclusion In this study, polyclonal S. Schwarzengrund strains of ST241 type were isolated from the patients. They were susceptible to common antibiotics, but carried the virulence genes contained in SPI1-5 and CdtB-islet coding genes and might cause an outbreak of infection. Attention should be paid to the tendency and threat of clinical S. Schwarzengrund infection and continuous surveillance and investigation should be performed.
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Objective To investigate antimicrobial resistance and ceftriaxone resistance mechanisms in clinically isolated nontyphoidal Salmonella(NTS),and provide evidence for the prevention and control of NTS infection and rational use of antimicrobial agents.Methods 108 NTS isolates were isolated from stool specimens of outpatients with acute diarrhea in the Second Hospital of Tianjin Medical University and Tianjin Medical University General Hospital from May to October of 2014,NTS were performed antimicrobial susceptibility testing;non-ceftriaxone-susceptible isolates were typed by serological,multilocus sequence (MLST),and pulsed-field gel electrophoresis (PFGE)methods,extended-spectrumβ-lactamase (ESBL)detection and AmpC genes were detection.Results Among 108 NTS isolates,mono-drug resistance rate to 11 antimicrobial agents was 49.07% (n= 53),multidrug resistance rate was17.59% . Susceptibility rates to nalidixic acid,levofloxacin,ciprofloxacin,ceftriaxone,and ertapenem were 61.11% ,66.67% ,68.52% ,97.22% ,and 100.00% respectively. Three non-ceftriaxone-susceptible NTS isolates were detected,2 were ST11 Salmonellaenterica serotype (Sa8709,Sa8771),1 was ST34 Salmonellatyphimurium serotype(Sa8763). Cluster analysis of PFGE revealed that Sa8709 was highly similar to Sa8771 strains(91 .70% ), but the similarity to Sa8763 was low(55.80% );Sa8709 strain carried CTX-M gene,Sa8771 strain carried CTX-M and TEM genes,Sa8763 strain carried OXA gene. Conclusion Clinically isolated NTS in this area are low resistant to fluoroquinolones,multidrug resistant strains carrying ESBLs have emerged.
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Objective To develop anoverlap syndrome (OS)rat model by intermittent hypoxia (IH) exposure on the base of pre-existing emphysema, and to explore its characters of severe systemic inflammation and immune responses. Methods Sixty Wistar rats were put into four groups:control group, IH group, emphysema group and overlap (emphysema+IH) group. The peripheral blood samples were collected for detecting apoptosis of CD3+CD4+,CD3+CD8+T lymphocytes and neutrophils (PMN). Tumor necrosis factor (TNF)-αand interleukin (IL)-6 were evaluated by ELISA. The bronchoalveolar la-vage fluid (BALF) was taken to calculate the ratio of macrophages, neutrophils and lymphocytes under light microscope. Tis-sue blocks of lung, liver, pancreas, and right carotid artery were taken for pathologic scoring. Results The apoptotic rates of PMN and CD3 +CD8 +T cells were significantly lower in overlap group than those of other three groups (P<0.05). Pro-in-flammatory factor IL-6, TNF-αand peripheral blood CD3 +CD4 +T cell apoptosis were the highest in overlap group com-pared to those of other three groups (P<0.05). The ratio of PMN and macrophages in BALF were significantly higher in em-physema group than those of other three groups (P<0.05) and the pathology scores of lung, liver, pancreas, the ratio of carot-id artery intima-media thickness of whole thickness of vascular were significantly higher in overlap group than those of other three groups (P < 0.05).Conclusion In rat model of intermittent hypoxia-emphysema there are more serious systemic multi-organ inflammation and immune responses.
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Objective To compare the solution rate of berberine in Gegen Qinlian decoction and Chinese goldthread decoction,to clarify the scientific nature of the traditional Chinese medicine compound.Methods High performance liquid chromatography(HPLC)was used to detect the content of berberine in Gegen Qinlian decoction and Chinese goldthread decoction.Chromatographic condition was C1 8 chromatographic column,mobile phase was acetonitrile:0.05mol/L of potassium dihydrogen phosphate(48:52)(0.4g sodium dodecyl sulfate was added to per 1 00mL solution,adjusted pH to 4.0 with glacial acetic acid),detection wavelength was 345nm,the sample size was 1 0μL,flow rate was 1 .0mL/min,column temperature was 30℃.Results The average decoct rate of berberine in Gegen Qinlian decoction was 0.73%,while that was 0.45% in Chinese goldthread decoction,there was significant difference between the two groups(P <0.01 ).Conclusion The efficacy of TCMcompound Gegen Qinlian decoction is greater than the efficacy of the unilateral simple accumulation,so traditional Chinese medicine compound has certain scientific nature.
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Objective To study the expression of active efflux pump AdeABC,AdeIJK,AdeFGH,AbeM,AbeS,CraA,MdtL in clinical Acinetobacter baumannii isolates and whether the efflux pumps confers resistance to antibiotics.Methods Thirty-two multi-drug resistant Acinetobacter baumannii islates and 10 sensitive isolates were collected.Genes of the exporter protein were amplified by PCR.The expression of adeB,adeJ,adeG,abeM,abeS,craA,mdtL were examined by real-time fluorescence quantitative RT-PCR.The controlling genes adeRS and adeL were amplified by PCR and sequenced.Results The positivity rates of adeB,adeJ,adeG,abeM,abeS,craA,mdtL were 100％,100％,100％,96.88％,100％,100％ and 93.75％ respectively in 32 multi-drug resistant Acinetobacter baumannii isolates,and all 100％ in 10 sensitive isolates.The difference of expression of adeB,abeM and mdtL were significant( P＜0.001,P =0.001,P=0.013) between 21 multi-drug resistant isolates of C clone and 10 sensitive isolates.The mutations of adeRS existed in 2 multi-drug resistant isolates,no point mutation of adeL.Conclusion The expression of AdeABC,AbeM and MdtL may involved in the resistant mechanisms of the clinical Acinetobacter baumannii islates.
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ObjectiveTo determine the allele frequency and genotypic distribution of plateletactivating factor acetylhydrolase (PAF-AH) gene polymorphism in the patients with sepsis. Methods Ala379Val,Val279Phe site genotypes were determined in patients (n=66) and healthy controls(n=68) by means of restriction fragment length polymorphism analysis of polymerase chain reaction products,DNA sequencing was used to detect the PCR product containing allele gene polymorphism.SPSS13.0 statistical software was use to analyze.Results All the samples by PCR-RFLP analysis of the PAF-AH gene in the Ala379Val site have three kinds of genotypes: in 66 cases of sepsis group there were 1 homozygous Val/Val type,19 heterozygous Val/Ala type,46 homozygous Ala/Ala type.In 68 cases of control group there were 2 Val/Val type,22 Val/Ala type,44 Ala/Ala type.The Ala379Val allele frequency and genotypic distribution in the patients with sepsis was not significantly different from those in the healthy controls.No statistically significant difference was observed between the survival group and the death group ( P＞0.05 ).PAFAH gene of Va1279Phe polymorphism could have three kinds of genotypes.All 66 patients in the sepsis group were the homozygous Val/Val type.Control group 68 cases,only one case was homozygous Phe/Phe type,and the others were homozygous Val/Val type,not found heterozygous Val/Phe type.The Val279Phe genotypic distribution and allele gene frequency in the patients with sepsis was not significantly different from those in the healthy controls; no statistically significant difference was observed no statistically significant difference was observed between the survival group and the death group ( P＞0.05 ).ConclusionNo associations were found between PAF-AH gene Ala379Val and Val279Phe polymorphisms and sepsis susceptibility,prognosis and severity.
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Objective To survey birth defects of neonates conceived by using various types of in vitro fertilization and embryo transfer (IVF-ET) between 1998 and 2007 in Shanghai. Methods From 1998 to 2007, 8507 neonates from 6551 pregnancies conceived through assistant reproductive technology (ART) from 7 reproductive medicine center in Shanghai were enrolled in this retrospective study, including Shanghai Ji-Ai Genetics and IVF Institute, Shanghai Jiaotong University School of Medicine affiliated Renji Hospital,Buijin Hospital, China Welfare Institute International Maternal and Infant Health Hospital, Shanghai First Maternity and Infant Health Hospital, Shanghai the Ninth People's Hospital and the Second Military Medical University affiliated Changhal Hospital. The clinical data about the type and incidence of birth defect were analyzed. Meanwhile, the factors associated with birth defect were investigated, such as various ART,gender, mother age, numbers of gestational sac, the source and quality of sperm. Results The birth defect rate was 1.08% (92/8507) and the most remarkable malformation occurred in circulation system [34% (31/92)]. The rates of major congenital malformations were 1.21% (34/2799) in fresh conventional IVF-ET, 1.07% (20/1871) in IVF-frozen-thawed embryo transfer(IVF-FET), 1.04% (23/2212) in fresh intracyto plasmic sperm injection (ICSI)-ET and 0.92% (15/1625) in ICSI-FET, which did not show statistical difference (P>0.05). There was also no significantly different incidence of malformations between 1.12% (49/4371) in male neonates and 1.02% (42/4136) in female neonates (P >0.05).However, the occurrence of congenital malformation is positively related with maternal age, the rates were 0.84% (41/4884) in mother's age less than 30 years and 1.77% (16/902) in mother' age more than 35 years, which reached statistical difference (P < 0.05). It also showed remarkable different incidence among 0.53% (25/4679) in singletons, 1.59% (57/3576) in twins and 3.97% (10/252) in triplets (P< 0.05). No remarkable difference of malformation rate among sperm sources used in fertilization were observed, including 1.09% (81/7419)in ejaculated sperm, 1.21% (7/579)in percutaneous epididymal aspiration (PESA) and 0. 79% (4/509) in donor sperm (P>0.05). Conclusions The overall incidence of birth defect in neonates from ART is similar to those conceived naturally and is not associated with different types of IVF-ET, embryo cryopreservation, sperm sources. However, maternal age and multiple pregnancies coder the higher possibility of birth defect.
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Objective To study the expression of active efflux pump AdeABC in clinical Acineto-bacter baumannii islates and whether this efflux pump confers resistance to antibiotics. Methods The anti-biotic susceptibility and the function of efflux pump inhibitor were tested by micro-dilution broth method. The expression of adeB was examined by RT-PCR. The controlling gene adeRS was amplified by PCR and se-quenced. Results Thirty multidrug resistance Acinetobacter baumannii isolates and 5 sensitive isolates for PCR were both abtained the expected products of adeB and adeRS. The mRNA expression of adeB in 15 multidrug resistance(MDR) isolates were positive, but there was no expression of adeB in 5 sensitive iso-lates. The mutations of adeRS existed in 2 MDR isolates. Conclusion The expression of AdeABC may in-volved in the resistant mechanisms of the clinical MDR Acinetobacter baumannii isolates.
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OBJECTIVE To investigate the repressor SmeT and other sequences of efflux pump SmeDEF in the clinic isolates of Stenotrophomonas maltophilia to involve in antibiotic resistance. METHODS The mRNA level of smeD examined the resistance profile of the clinical strains to antimicrobials was by agar dilution and their reaction to pump inhibitor was investigated by RT-PCR,and the smeD-smeT fragment was amplified by PCR,and then sequenced.Adding poly(A) to 3′ end of RNA,applying 3′RACE,nested PCR,then sequence-analysis. RESULTS The amino acid sequences of the forepart of SmeT were conservative in all the 7 strains.In antibiotic-resistant strains with positive reaction to pump inhibitor,the intergenic sequences of smeD-smeT were different from those in sensitive ones.The variations in(-165 to-82)nt of smeT and several nucleic acids before the start codon of smeT,possibly involved in antimicrobial resistance.All isolates didn′t have Leu166Gln mutation within the SmeT protein of D457R,which was reported possibly associated with antibiotic resistance.The resistant strains with efflux positively inhibited had Asp218Glu substitution,different from the negatively-inhibited ones. CONCLUSIONS The expression level of SmeDEF is related with the antibiotic resistance of S.maltophilia.The possible resistance mechanism includes the variations in the intergenic smeD-smeT and/or smeT codon sequence.