Objective To explore the characteristics of intestinal Microbiota in T2DM patients by two molecular fingerprint technologies,and investigate the correlation of intestinal microbiota and T2DM,and evaluate the application value of two fin-gerprint technologies.Methods Fecal samples of 8 healthy groups and 7 diabetes patients were collected.Then the total DNA of gut microbiota was extracted.Through the analysis of products by two molecular fingerprints of ERIC-PCR and DGGE-PCR,ecological characteristics of diversity and similarity of gut microbiota were obtained in healthy groups and dia-betes patients.Results Compared to healthy groups,the number of bands and Shannon-Wiener index of gut microbiota in di-abetes patients was decreased but no statistical significance.The similarity in patients group was declining(P <0.05),and the construction of gut microbiota was inclined to differ.Two fingerprint technologies of ERIC and DGGE could directly re-flect the diversity of gut microbiota and were the modern molecular biological techniques without depending on cultivation. ERIC was simple and convenient,had a better reflection of microbial diversity,but gel band cutting and regarded asa proper approach with higher diffraction efficiency and excellent repetition to studysequencing couldn’t be performed since there were more influencing factors on the experiment.DGGE could better reflect the ecological characteristics such as microbial diversity and similarity,and selecting bands,gel band cutting and sequencing could be done.Conclusion The composition and construction of gut microbiota in diabetes patients were changed,which suggests the occurrence of the disease had the correlation with gut microbiota.ERIC and DGGE is regarded as a proper approach with higher diffraction efficiency and ex-cellent repetition to study intestinal microbiota,but also gel band cutting,sequencing,bacteria identification can be performed by DGGE,both can be used in combination.

Abstract: Pneumococcal vaccine plays a key role in preventing diseases such as pneumonia and meningitis caused by Streptococcus pneumoniae (S. pneumoniae). Capsular polysaccharide, C-polysaccharide and phosphorus content are important indicators for quality control of polysaccharide antigens in vaccine development and production. In this study, a quantitative 1H NMR and 31P NMR method based on a single internal standard hexamethylphosphoramide (HMPA) was developed to achieve simultaneous determination of capsular polysaccharide, C-polysaccharide and phosphorus content in 6A, 6B, 18C, 19A, 19F and 23F S. pneumoniae polysaccharide antigens. Using the internal reference comparison method, the effect of solubility of polysaccharide on quantitative 1H NMR determination was investigated. It was found that the determination results of quantitative 1H NMR were affected by the viscosity and concentration of polysaccharide solution. It was found that high viscosity polysaccharides at 3−15 mg/mL and low viscosity polysaccharides at 5−25 mg/mL were the optimal detection solution concentration range. This “one internal standard three quantitative” NMR method has good precision, specificity and accuracy, and provides a valuable new strategy for the quality control of pneumococcal vaccine.

Objective:To explore the effect of augmented renal clearance(ARC)on 24-hour area under the concentration-time curve to minimum inhibitory concentration ratio(AUC 24/MIC)of vancomycin and prognosis in critical children, thus to provide proposal for individual dosage regimen. Methods:Sixty-five critical children treated with vancomycin, who suffered from sepsis/septic shock, were brought into this retrospective cohort study.According to estimate glomerular filtration rate, these children were divided into ARC group ( n=27) and normal group ( n=38). The influencing factor of AUC 24/MIC of vancomycin and therapy prognosis for two groups were detected and analyzed. Results:There were no significant differences between two groups in basic setting (age, sex, weight), scores of pediatric sequential organ failure assessment and pediatric risk of mortality Ⅲ, infection markers (C-reactive protein and procalcitonin), glutamic-pyruvic transaminase, hypoproteinemia, usage of diuretic and vasoactive agent( P>0.05). The patients from ARC group showed lower levels than those from normal group in AUC 24/MIC of vancomycin[375.2(300.8, 489.4) vs. 443.6(412.3, 593.2), Z=2.263, P=0.024] and it′s target achievement ratio (TAR)(40.7% vs. 76.3%, χ2=8.440, P=0.005). When usage of diuretic and vasoactive agent, the AUC 24/MIC of ARC group was lower than that of normal group( P<0.05). But there was no significant difference between ARC group and normal group regarding hypoproteinemia( P>0.05). The days of body temperature steady at least 48 hours[7.0(5.5, 9.0)d vs. 6.0(5.0, 8.0)d], the length of hospital stay[39.0(21.0, 58.0)d vs. 20.5(16.0, 28.0)d], the length of PICU stay[14.0(9.0, 31.5)d vs. 10.0(5.0, 15.0)d] were longer than those in normal group( P<0.05). There were no significant differences between ARC group and normal group regarding days of ventilation and infectious markers decreased at least 50%, as well as 28-days mortality( P>0.05). The multivariable analysis showed that the presence of ARC, hypoproteinemia, use of diuretics and vasoactive agent were significantly associated with AUC 24/MIC of vancomycin( P<0.05). Conclusion:ARC may down regulate levels of AUC 24/MIC and TAR of vancomycin.During ARC period, the usage of diuretic and vasoactive agent could affect the AUC 24/MIC of vancomycin.Individual dosage regimen should be employed for critical children suffered with ARC.

Objective To evaluate the antibacterial effect of mononuclear cells(MCs)in the liver lavage solution. Methods For in vitro experiment, MCs were col ected from the liver lavage solution of SD rats and divided into the supplement of interleukin(IL)-15 and non-supplement groups. The MCs were co-cultured with Pseudomonas aeruginosa (P. aeruginosa)for 4 h and then the supernatant was collected and MCs were lysed. The bacterial load in the lysate was detected after LB plate culture. The levels of interferon(IFN)-γand tumor necrosis factor(TNF)-αin the supernatant were measured by enzyme-linked immune absorbent assay(ELISA). For in vivo experiment, 40 SD rats were administered via tracheal injection of P. aeruginosa solution at a dose of 1×109 CFU/mL and randomly divided into four groups(n=10).In the control group, physiological saline was given via gavage. In the immunosuppression group, tacrolimus(FK506)was delivered via gavage. In the MC group, MCs at a dose of 1.0×108 was given via intravenous injection after use of FK506. In the IL-15 pretreated-MC group, IL-15 pretreated-MCs at a dose of 1.0×108 were administered via intravenous injection after application of FK506. The lavage solution of pulmonary alveolus and the rat lung tissue were col ected. The bacterial load was detected after LB plate culture. The expression of IFN-γand TNF-αin the pulmonary alveolus and lung tissue were measured by ELISA and Western blot. Results Compared with MCs alone, IL-15 pretreated-MCs exhibited significantly higher antibacterial capability in vitro. The CFU was 35%of untreated MCs. The synthesis and release capabilities of IFN-γandTNF-αweresignificantlyenhanced.Comparedwiththecontrolgroup,thequantityofimmunecelsinthelungtissuewas decreased and the bacterial load in the lung tissue and the lavage solution of pulmonary alveolus was significantly increased, whereas the expression levels of IFN-γand TNF-αtended to decline in the immunosuppression group. Administration of IL-15 pretreated-MCs significantly enhanced the quantity of immune cel s in the lung tissue, decreased the bacterial load and increased the secretion of IFN-γand TNF-α. Conclusions MCs in the liver lavage solution exhibit favorable antibacterial activity.Underimmunosuppressioncondition,thedefensecapabilityofthehostagainsttheopportunisticpathogenicbacteria is significantly enhanced.

Objective: To know the genetic characteristics of nucleoprotein(N)of wild type measles virus in Xi’an, 2013-2017. Methods: We used the pharyngeal swab specimens of suspected measles cases which were tested positive for measles virus (MV) nucleic acid by Real-time RT-PCR to isolate MV strains with Vero-SLAM cells, extracted RNA of the strains, amplified the N genes of MV strains and the pharyngeal swab specimens which had lower Ct values by One-Step RT-PCR, and analyzed the result after sequencing. Results: Thirty carboxyl terminal sequences of the N genes of MV endemic strains were obtained and identified as genotype H1a. The nucleotide and amino acid homology among the epidemic strains were 97.3%-100% and 96.7%-100%, among the epidemic strains and S191 were 91.3%-92.2% and 87.3%-90.0%, while among the epidemic strains and C47 were 92.2%-92.7% and 88.0%-90.7%. The nucleotide homology among the epidemic strains and wild strains of same period in Anhui, Gansu, Shandong, Henan, Jilin were higher than that in Shanxi, Sichuan, Hunan and Guangdong. The most different amino acid sites among the epidemic strains and the two vaccine strains which had high level of entropy were 422, 457 and 497, secondly 467, thirdly 500. The mutations at the 467 site only existed in 2017. dN/dS=0.199 and no positive selection site was found. Conclusions The dominant genotype of the epidemic MV strains in Xi’an was H1a in 2013-2017, the genetic diversity of viruses decreased in 2016-2017. We should strengthen the surveillance of etiology of MV to provide evidence for elimination of measles.