Establishment and Modification of Ninety-seven Pneumococcal Serotyping Assays Based on Quantitative Real-time Polymerase Chain Reaction / 生物医学与环境科学（英文）

OBJECTIVE@#To establish and modify quantitative real-time polymerase chain reaction (qPCR)-based serotyping assays to distinguish 97 pneumococcal serotypes.@*METHODS@#A database of capsular polysaccharide ( cps) loci sequences was generated, covering 97 pneumococcal serotypes. Bioinformatics analyses were performed to identify the cps loci structure and target genes related to different pneumococcal serotypes with specific SNPs. A total of 27 novel qPCR serotyping assay primers and probes were established based on qPCR, while 27 recombinant plasmids containing serotype-specific DNA sequence fragments were constructed as reference target sequences to examine the specificity and sensitivity of the qPCR assay. A panel of pneumococcal reference strains was employed to evaluate the capability of pneumococcal serotyping.@*RESULTS@#A total of 97 pneumococcal serotyping assays based on qPCR were established and modified, which included 64 serotypes previously reported as well as an additional 33 serotypes. Twenty-seven novel qPCR serotyping target sequences were implemented in the pneumococcal qPCR serotyping system. A total of 97 pneumococcal serotypes, which included 52 individual serotypes and 45 serotypes belonging to 20 serogroups, could not be identified as individual serotypes. The sensitivity of qPCR assays based on 27 target sequences was 1-100 copies/µL. The specificity of the qPCR assays was 100%, which were tested by a panel of 90 serotypes of the pneumococcal reference strains.@*CONCLUSION@#A total of 27 novel qPCR assays were established and modified to analyze 97 pneumococcal serotypes.

A New High-throughput Real-time PCR Assay for the Screening of Multiple Antimicrobial Resistance Genes in Broiler Fecal Samples from China / 生物医学与环境科学（英文）

OBJECTIVE@#Antimicrobial resistance (AMR) has become a global concern and is especially severe in China. To effectively and reliably provide AMR data, we developed a new high-throughput real-time PCR assay based on microfluidic dynamic technology, and screened multiple AMR genes in broiler fecal samples.@*METHODS@#A high-throughput real-time PCR system with an new designed integrated fluidic circuit assay were performed AMR gene detection. A total of 273 broiler fecal samples collected from two geographically separated farms were screened AMR genes.@*RESULTS@#The new assay with limits of detection ranging from 40.9 to 8,000 copies/reaction. The sensitivity rate, specificity rate, positive predictive value, negative predictive value and correct indices were 99.30%, 98.08%, 95.31%, 99.79%, and 0.9755, respectively. Utilizing this assay, we demonstrate that AMR genes are widely spread, with positive detection rates ranging from 0 to 97.07% in 273 broiler fecal samples. blaCTX-M, blaTEM, mcr-1, fexA, cfr, optrA, and intI1 showed over 80% prevalence. The dissemination of AMR genes was distinct between the two farms.@*CONCLUSION@#We successfully established a new high-throughput real-time PCR assay applicable to AMR gene surveillance from fecal samples. The widespread existence of AMR genes detected in broiler farms highlights the current and severe problem of AMR.

Differential protein expression in patients with urosepsis / 中华创伤杂志(英文版)

PURPOSE@#Urosepsis in adults comprises approximately 25% of all sepsis cases, and is due to complicated urinary tract infections in most cases. However, its mechanism is not fully clarified. Urosepsis is a very complicated disease with no effective strategy for early diagnosis and treatment. This study aimed to identify possible target-related proteins involved in urosepsis using proteomics and establish possible networks using bioinformatics.@*METHODS@#Fifty patients admitted to the Urology Unit of Lanzhou General PLA (Lanzhou, China), from October 2012 to October 2015, were enrolled in this study. The patients were further divided into shock and matched-pair non-shock groups. 2-DE technique, mass spectrometry and database search were used to detect differentially expressed proteins in serum from the two groups.@*RESULTS@#Six proteins were found at higher levels in the shock group compared with non-shock individuals, including serum amyloid A-1 protein (SAA1), apolipoprotein L1 (APOL1), ceruloplasmin (CP), haptoglobin (HP), antithrombin-III (SERPINC1) and prothrombin (F2), while three proteins showed lower levels, including serotransferrin (TF), transthyretin (TTR) and alpha-2-macroglobulin (A2M).@*CONCLUSION@#Nine proteins were differentially expressed between uroseptic patients (non-shock groups) and severe uroseptic patients (shock groups), compared with non-shock groups, serum SAA1, APOL1,CP, HP, SERPINC1and F2 at higher levels, while TF, TTR and A2M at lower levels in shock groups.these proteins were mainly involved in platelet activation, signaling and aggregation, acute phase protein pathway, lipid homeostasis, and iron ion transport, deserve further research as potential candidates for early diagnosis and treatment. (The conclusion seems too simple and vague, please re-write it. You may focus at what proteins have been expressed and introduce more detail about its significance.).

Dissemination of insertion sequence common regions 1 and int1 gene and drug resistance of 483 Escherichia coli and Klebsiella pneumonia broiler isolates / 中华预防医学杂志

Objective@#To investigate and analyze distribution characteristics of two multidrug resistance related genes in broiler isolates in Shandong province.@*Methods@#The pre slaughter broilers were chosen from Shandong province in this study in June, 2014. A total of 400 fecal samples from five different zones (east, south, west, north and middle) of the hen house were collected. 373(77.2%) Escherichia coli and 110 (22.8%) Klebsiella pneumonia strains were isolated, and ISCR1 and int1 gene were detected by PCR assay and sequencing. The resistance to 10 drugs belonging to 8 classes antimicrobial drugs were obtained by using minimal broth dilution method and data analysis. The difference between isolates and drug resistance profiles was analyzed.@*Results@#Among 483 isolates, 440 isolates (91.1%), 126 isolates (26.1%) and 126 isolates (26.1%) were detected as int1, ISCR1 and both two gene carriers, respectively. The rate of 37 E. coli isolates not carried ISCR1 or int1 gene resistant to 0 to 2, 3 to 5, 6 to 8 classes antimicrobial agents was 13.5% (n=5), 78.4% (n=29), and 8.1% (n=3), respectively; the rate of 288 only int1 gene E. coli carriers resistant to 0 to 2, 3 to 5, 6 to 8 groups antimicrobial agents was 2.4% (n=7), 74.7% (n=215), and 22.9% (n=6), respectively. The data above showed significant difference (P<0.001). The rate of 26 only int1 gene K. pneumonia carriers resistant to 0 to 2, 3 to 5, 6 to 8 classes antimicrobial agents was 11.5% (n=3), 76.9% (n=20), and 11.5% (n=3), respectively; the rate of 78 both two gene K. pneumonia carriers resistant to0 to 2, 3 to 5, 6 to 8 groups antimicrobial agents was 0, 35.9% (n=28), and 64.1% (n=50), respectively. The data above showed significant difference (P<0.001).@*Conclusion@#Gene int1 and ISCR1 showed high prevalence in E. coli and K. pneumonia isolates. High level multi-drug resistance profile could be mediated by int1 and ISCR1 gene co-existence.

Study on virulence factors of Candida tropicalis isolated from clinical samples / 中华流行病学杂志

Objective To determine the in vitro production of virulence factors for Candida (C.) tropicalis,including aspartyl proteinases,phospholipases and hemolytic activities,describe the regulation of virulence factors varying with time in C.tropicalis,and analyze the differences in aspartyl proteinases and hemolytic activities of C.tropicalis isolated from anatomically distinct sites.Methods A total of 64 C.tropicalis strains were spot-inoculated onto bovine albumin agar,egg yolk agar and sheep blood agar plates,respectively.Then the plates were incubated for 24,48 and 72 hour at 37 ℃,respectively.The aspartyl proteinases,phospholipase and hemolytic activities were determined at each time point,respectively.Results All the C.tropiclais isolates showed positive aspartyl proteinases and hemolytic activities at each time point,but no phospholipases activity was detected in C.tropicalis.On comparison of aspartyl proteinases and hemolytic activities at different time points,aspartyl proteinases activity at 48 and 72 hour was higher than that at 24 hour.During 72 hour,hemolytic activity of C.tropicalis increased.No statistical significant differences in aspartyl proteinases and hemolytic activities of C.tropicalis were observed among different infection sites (P=0.368 and 0.985).Conclusion The C.tropicalis clinical isolates in China have aspartyl proteinases activity,hemolytic activity,but have no phospholipase activity.

Study on genotype and virulence of Cryptococcus neoformans and Cryptococcus gattii clinical isolates in Guigang, Guangxi Zhuang Autonomous Region / 中华流行病学杂志

<p><b>OBJECTIVE</b>To understand the species, genotypes and mating types of Cryptococcus neoformans and Cryptococcus gattii isolated from clinical samples in Guigang, Guangxi Zhuang Autonomous Region.</p><p><b>METHODS</b>A total of 20 Cryptococcus strains were isolated from clinical samples in Guigang from 2009 to 2012. The biological identification was conducted by polymerase chain reaction (PCR) to amplify internal transcribed spacer (ITS) sequences. The serotypes and mating types of C. neoformans and C. gattii were identified by PCR with serotype-specific and mating type-specific primers. The genotype was characterized by PCR fingerprinting and URA5 gene restriction fragment length polymorphism (URA5-RFLP). Phenotype study included growth test at 37 °C, melanin production test and urease test.</p><p><b>RESULTS</b>Among the 20 strains, 19 (95%) were identified as C. neoformans varieties (var.) grubii (serotype A, mating type α, genotype VN I), and only 1 was identified as C. gattii (mating type α, genotype VG I). The results of virulence test showed that all the strains grew well at 37 °C and positive in both urease test and melanin production test.</p><p><b>CONCLUSION</b>C. neoformans var. grubii (serotype A, genotype VN I and mating type α) was the predominant pathogen causing cryptococcosis in Guigang, and C. gattii strain was also detected.</p>

A study on human umbilical vein endothelial cell ECV304 proliferation induced by Saccharomyces albicans / 华西口腔医学杂志

<p><b>OBJECTIVE</b>To study the effects of Saccharomyces albicans (S. albicans) on the cell cycle distribution and proliferation of human umbilical vein endothelial cell ECV304 cells in vitro.</p><p><b>METHODS</b>The line of ECV304 cultured in vitro were divided into four groups which were treated by S. albicans supernatant, S. albicans inactivated bacilli, supernatant and inactivated bacilli mixture, normal culture medium. The proliferous effect of ECV304 induced by supernatant, inactivated bacilli, supernatant and inactivated bacilli mixture using the methods of MTT, cell count, microscope and flow cytometry were conducted.</p><p><b>RESULTS</b>In the condition of different times and different culture concentrations, ECV304 cells incubated with 4-fold diluted S. albicans supernatant for 48 h increased the proliferation rate. The S and G2/M population of ECV304 cells increased after incubated with S. albicans supernatant for 40 h, which showed significant increasing cell proliferation index (PI) (P < 0.05). The PI of the cells treated by inactivated bacilli showed no significant change (P > 0.05).</p><p><b>CONCLUSION</b>S. albicans could induce ECV304 cell proliferation which depends on the release of metabolic products of S. albicans.</p>

Mutation analysis of the eda-A1 gene for hypohidrotic ectodermal dysplasia and construction of recombined eukaryotic expression vector / 华西口腔医学杂志

<p><b>OBJECTIVE</b>The purpose of this study was to clone and analyze mutation in the eda-A1 gene for hypohidrotic ectodermal dysplasia (HED), and to construct a new recombined eukaryotic expression vector (mutant M, wild W) as a basis for further study on the genetic function.</p><p><b>METHODS</b>After total mRNA was extracted from peripheral blood lymphocytes from the HED affect patient and control, eda-A1 gene was amplified by reverse transcription polymerase chain reaction (RT-PCR) with a pair of specific primers containing the constriction enzyme sites of BamH I and Hind III. When the vector pcDNA3.1(-) and eda-A1 (M/W) were digested by BamH I and Hind III respectively, eda-A1 (M/W) fragment was then ligated to vector pcDNA3.1 (-) and the new vector was named as pcDNA3.1 (-)-eda-A1-M/W.</p><p><b>RESULTS</b>eda-A1 gene was successfully cloned and a novel missence mutation was identified, which changes the codon 306 from glutamine to proline. PCR, restrictive endonuclease analysis and DNA sequencing were then performed to identify the recombinant eukaryotic expression vector pcDNA3.1 (-)-eda-A1-M/W, and the results were surely confirmed.</p><p><b>CONCLUSION</b>Our result indicates that the novel missense mutation in eda is associated with the isolated tooth agenesis and provide preliminary explanation for the abnormal clinical phenotype at a molecular structural level. And also, the recombinant eukaryotic expression vector pcDNA3.1 (-)-eda-A1-M/W was successfully constructed, which will be thereafter taken use of further study on eda gene in odontogenesis.</p>

Comparative Study of Oxygen and Pressure Support Therapy on Plateau Hypoxia at an Altitude of 3992 Meters / 中国呼吸与危重监护杂志

Objective To compare the effects of oxygen therapy and local pressurization in alleviating plateau hypoxia at high altitude.Methods Forty-five healthy male soldiers were investigated at an altitude of 3992 meters.The subjects were randomly divided into three groups, ie.an oxygen inhalation group, a single-soldier oxygen increasing respirator (SOIR) group and a BiPAP group.The oxygen inhalation group was treated with oxygen inhalation via nasal catheter at 2 L/min.SOIR was used to assist breath in the SOIR group.The BiPAP group were treated with bi-level positive airway pressure ventilation, with IPAP of 10 cm H20 and EPAP of 4 cm H2O.PaO2、PaCO2、SpO2 and heart rate were measured before and 30 minutes after the treatment.Results There were continuous increase of PaO2 from (53.30±4.88) mm Hg to (58.58±5.05) mm Hg and (54.43±3.01) mm Hg to (91.36±10.99) mm Hg after BiPAP ventilation and oxygen inhalation, respectively (both P < 0.01).However, the PaO2、of the SOIR group was decreased from (56.00±5.75) mm Hg to (50.82±5.40) mm Hg (P < 0.05).In the other hand, the PaCO2、 was increased from (30.41±1.51) mm Hg to (32.5±2.98) mm Hg in the oxygen inhalation group (P< 0.05), declined from (28.74±2.91) mm Hg to (25.82±4.35) mm Hg in the BiPAP group (P < 0.05), and didn't change significantly from (28.65±2.78)mm Hg to (29.75±3.89) nun Hg in the SOIR group (P > 0.05).Conclusions Both BiPAP ventilation and oxygen inhalation can alleviate plateau hypoxia by improving PaO2 at 3992 meter altitude while SOIR has no significant effect.

Preparation of atenolol monolithic osmotic pump tablets / 药学学报

<p><b>AIM</b>The simplified preparative method and formulation for atenolol monolithic osmotic pump tablets were investigated.</p><p><b>METHODS</b>The core tablets with an indentation were compressed by the punch with a needle. Osmotic pump tablets were prepared by coating the indented tablets. Similarity factor was used to evaluate formulation of osmotic pump tablets.</p><p><b>RESULTS</b>The formulation of core tablets and the composition and thickness of coating membrane showed marked effects on drug release. Orifice size of core tablets in the range of 1.00 - 1.14 mm scarcely affected drug release.</p><p><b>CONCLUSION</b>The preparation of osmotic pump tablets was simplified with the exempting of laser drilling. The atenolol monolithic osmotic pump tablets could deliver drug constantly for 24 h.</p>

Treatment of 30 Cases of Vitiligo by Cupping Method plus External Application of Chinese Herbs / 针灸推拿医学（英文版）

Thirty patients with vitiligo were treated by cupping of skin lesion and compress of Chinese herbs, compared with western medical treatment. After 3-course's treatments the total effective rate ws 96.7% in treatment group and 76.7% in control group. There was a significant difference (P＜0.05)between the two groups shown by statistical analysis.