Objective To elucidate the deleterious diabetogenic effect of tacrolimus on Chinese Han people.Methods According to patients' whole blood trough levels,the concentration of tacroli- mus was 5,15 and 25?g/L.Human islet cells cultured in vitro were exposed to various concentra- tions of tacrolimus for 72 h respectively,then stimulated by glucose at low (2.8 mmol/L) and high (16.7 mmol/L) levels.Glucose-stimulating insulin secretion during subsequent static incubation was measured using the ultrasensitive human insulin ELISA kit.The assessment of islet cell viability was studied with acridine orange (AO)-propidium iodide (PI).According to fasting plasma glucose,461 subjects were divided into normal glucosetolerance group (FPG＜5.55 mmol/L),impaired fasting glu- cose group (5.55 mmol/L＜FPG＜6.88 mmol/L) and posttransplant diabetes mellitus group (FPG≥6.89 mmol/L).A retrospective review of 461 non-diabetic kidney recipients completing at least 12 months of follow-up was performed to determine risk factors,incidence,and characteristics of post- transplant diabetes mellitus (PTDM).Results In vitro,lower tacrolimus concentration had not dele- terious effect on insulin secretion and viability of islet;high tacrolimus impaired?-cell survival as well as insulin secretion.Of the 461 patients with no history of diabetes at transplantation over the study period of 12 months,123 received tacrolimus and 338 received cyclosporine A (CsA).For patients receiving tacrolimus the cumulative incidence of PTDM was 13.8%,compared with 7.7% for pa- tients receiving CsA,showing no significant difference between them.The reduced tacrolimus level within the therapeutic window resulted in an increase in better glucose metabolism.Older age,family history of diabetes,acute rejection episodes and pre-transplant impaired fasting glucose were identified as risk factors for PTDM.Conclusion The diabetogenic effect of TAC is dose-dependent,and in the majority is reversible.Low initial doses,more rapidly tapering and lower maintenance levels of tacroli- mus decrease the incidence of PTDM in patients receiving tacrolimus.

A novel nucleic acid amplification method,termed loop-mediated isothermal amplification(LAMP),which amplifies DNA with high specificity,efficiency,and rapidity under isothermal conditions,may be a valuable tool for the rapid detection of infectious diseases.This method employs a DNA polymerase that have activity of strand displacement DNA synthesis and a set of four specially designed primers that recognize a total of six distinct sequences on the target DNA.LAMP can amplify a few copies of DNA to 109 in less than an hour.The final products are stem-loop DNA with several inverted repeats of the target and cauliflower-like structures with multiple loops.A positive reaction would be shown as a ladder-like pattern in a gel electrophoresis analysis.Because of the advantage,the LAMP method will be widely applied to research of nucleic acid,clinical diagnosis of infectious diseases and detection of genetically modified organisms etc.

In this study, microcalorimetry was adopted to establish a novel method for detecting the hemagglutination process of Radix Isatidis (Banlangen in Chinese, BLG), and to evaluate the hemagglutination activity diversity of BLG from various habitats. The hemagglutination biothermokinetics curves of positive reagent (phytohemagglutinin, PHA) and 8 batches BLG from different regions of the hemagglutination with 20% rabbit erythrocyte were recorded by microcalorimetry, then biothermokinetics parameters were abstracted, the hemagglutination utility of samples were calculated and analyzed by principal component analysis (PCA) and cluster analysis (CA), meanwhile the results were authenticated by micro-plate agglutination. It showed that the hemagglutination was an exothermic reaction, the reaction rate constant (k: 0.039-73.6 min(-1)), maximum reaction power (Pmax: -1 140.2 - 988.2 microW) and reaction enthalpy (Hi: -529.9 - 717.9 microJ) had good linear correlation with BLG extraction concentration (0.2-1.0 g mL(-1), r > 0.97), and PCA showed Pmax (531-1 335 microW) and Ht (585.2-989.2 microJ) could represent the hemagglutination activity diversity of BLG samples, just confirming with the results of micro-plate agglutination (the agglutination dilution was 3-11 respectively). According to the hemagglutination utility, the BLG samples from Good Agriculture Practice (GAP) regions, main producing area and general regions could be clustered correctly; meanwhile, the biothermokinetics curves with perfect distinctive fingerprint and specificity could give out more information for the quality control and evaluation for BLG. In conclusion, the microcalorimetry method established for detecting the hemagglutination activity of BLG samples on rabbit erythrocyte is sensitive and reliable, and could be adopted as an effective technique in detection aggulatination precisely, quantitatively and consecutively; and provide a novel approach for examining and evaluating quality for Chinese herbal medicine with aggulatinative activity such as BLG.

To establish kinetic assay method for the analysis of hemolysis and to investigate dynamic hemolytic process of polysorbate 80. The UV-VIS spectrum of heme changes when hemoglobin is released continuously during the hemolytic process. Therefore, dynamic hemolytic curve was determined as a new way to characterize the kinetic process of interaction between polysorbate 80 and red blood cells. The effect of polysorbate 80 on blood cells could be perfectly investigated by the hemolytic dynamics. Dynamic hemolytic parameters of polysorbate 80 were calculated according to the hemolytic curves. The constants of hemolytic rate and maximum hemolytic rate of polysorbate 80 had fine linear relationships at the range of 1-20 mg x mL(-1) and 5-20 mg x mL(-1), respectively. In comparison with the present official method such as macroscopic observation and static spectrophotometric methods, kinetic spectrophotometry has the advantages of real time, on-line determination, sensitive, objective, good reproducibility and 2-dimensional information acquired. Therefore, as a biological technique, kinetic spectrophotometry could be applied to evaluate the quality of polysorbate 80 and to screen other solubilizing excipients.

Objective To profile the phenotype and pathogens of cutaneous and mucous mycoses in a dermatology outpatient clinic in Nanchang region. Methods A review was performed to assess cutaneous and mucous mycoses diagnosed in the dermatology outpatient clinic of Dermatology Hospital of Jiangxi Province from 2006 to 2008. The relationship of clinical phenotype and pathogens to season, patients' age and gender was analyzed. Results A total of 7251 cases were collected, and the ratio of male to female patients was 2.3: 1. The most prevalent mycoses included tinea cruris (2702, 37.1%), pityriasis versicolor (1505, 20.8%) and tinea manus (727, 10.0%). In total, 4953 fungal strains were isolated from all the patients except for those with pityriasis versicolor, of them, Trichophyton rubrum accounted to 69.9%, Candida to 20.4%, and Trichophyton violaceum to 4.5%. Season, patients' age and gender were found to be associated with clinical phenotypes and pathogens of mycoses. Conclusions In the dermatology outpatient clinic of Nanchang region, tinea cruris is the most common superficial fungal disease, with the predominant pathogen being Trichophyton rubrum. Trichophyton violaceum is the primary pathogen of tinea capitis, which is different from other reports.

OBJECTIVETo investigate the abnormal change of immune function in patients with Pi-Qi deficiency Syndrome, and to explore the genomic mechanism of its genesis by cDNA chip techniques.

METHODSThe cross probe was made by extracting and microamplifying the total RNA and mRNA of peripheral white blood cells (WBC) in healthy subjects and patients with chronic gastritis and ulcerative colitis, which were labeled by Cy3 and Cy5 respectively. Then equal quantity of the two labeled probes were mixed and hybridized with cDNA chip, fluorescent signal of the chips were scanned with scanner. Data obtained were analyzed for comparing the difference of the expressive levels of immune associated genome in peripheral WBC in healthy subjects with those in patients.

RESULTSExpressions of CD9, CD164, PF4 and RARB gene in WBC of patients, both gastritis and colitis, were down-regulated while those of IGKC, DEFA1 and GNLY were up-regulated.

CONCLUSIONThe genesis of Pi-Qi deficiency syndrome has its immune associated genomic basis, and the immune functions are disordered in patients with that syndrome.

Adult ; Colitis, Ulcerative ; genetics ; immunology ; Diagnosis, Differential ; Gastritis ; genetics ; immunology ; Genome ; Humans ; Male ; Medicine, Chinese Traditional ; Middle Aged ; Oligonucleotide Array Sequence Analysis ; Qi ; Splenic Diseases ; genetics ; immunology ; Yang Deficiency ; genetics ; immunology

Objective To evaluate the clinical application of enteral nutrition by nasojejunal tube insertion and by percutaneous endoscopic gastrostomy (PEG) in elderly patients.Methods A total of 65 elderly patients with dysphagia recruited at our department from January 2010 to November 2014 were divided into the nasojejunal tube feeding group (35 cases) and the PEG feeding group (30 cases).Differences between these two groups in nutritional indexes,immunological indexes,complications and mortality were analyzed retrospectively.Results Serum total protein,albumin and prealbumin and upper arm circumferences all increased after treatment with nasojejunal tube feeding or percutaneous endoscopic gastrostomy (P＞0.05).There was overall improvement in nutritional status,as assessed by Nutritional Risk Screening 2002 (NRS2002).Specifically,the before/one month-after-treatment ratio of scores was 3.72±0.91/1.90±0.61 (t=7.24,P＜0.01) for the nasojejunal tube feeding group and 3.52±1.23/2.02±0.53 (t=4.17,P＜0.01) for the PEG feeding group.Compared with NRS2002 scores at one month post-operation,further improvement was achieved at 3 months postoperation both for the nasojejunal tube feeding group (1.89±0.65,t=5.21,P＜0.01) and for the PEG feeding group (1.91±0.62,t=4.40,P＜0.01).There was no difference in the indexes of nutrition,immune status or mortality between the two groups (P＞0.05).Although improvement in CD3+,CD4+,CD8+,CD4+/CD8+,IgA,IgG,and IgM was seen in both groups after operation,the differences did not reach statistical significance (P＞0.05).The incidence of aspiration pneumonia was notably lower (P＜0.05) while the incidence of diarrhea was much higher (P＜0.05) in the nasojejunal tube feeding group than in the PEG feeding group at one month and three months.The two groups had similar causes of death and mortality rates.Conclusion Both nasojejunal tube and PEG feeding can improve the nutritional status of elderly patients with dysphagia.However,the choice for the route of nutrition should be individualized.

Objective To evaluate the relationship of preoperative neutrophil-lymphocyte ratio (NLR) with clinicopathological features and prognosis of colorectal cancer in middle-aged and elderly patients.Methods A retrospective analysis was performed in 212 patients with colorectal cancer in the First Affiliated Hospital of Nanjing Medical University from January 2011 to June 2013.All patients were divided into middle-aged group (46-65 year old,n=130) and old-aged group (66-89 year old,n=82),The optimal cut-off point of NLR was identified by the area under receiver operating characteristic curve,while NLR ＞ 3.13 and NLR≤3.13 were classified as high and low NLR group.The clinicopathological features and prognosis between the two groups were compared.Results There was no difference in gender,tumor growth site,depth of invasion,tumor embolus,lymphatic metastasis,distant metastasis,TNM stage between low and high NLR group (allP＞ 0.05).However,the difference between high NLR group and low NLR group in old-aged group with diabetes mellitus was statistically significant (P＜0.05).The 1-,2-,and 3-year survival rate of the overall 212 patients were 96.2％ (204/212),87.7％ (186/212) and 74.5％ (158/212) In middle-aged group,the 1-,2-,and 3-year survival rates were 98.8％ (85/86),90.7％ (78/86) and 84.9％ (73/86) respectively in low NLR group,but 95.5％ (42/44),84.1％ (37/44) and 72.7％ (32/44) respectively in high NLR group,(allP＜0.05).In old-aged group,the 1-,2-,and 3-year survival rates were 95.7％ (44/46),89.1％ (41/46) and 73.9％ (34/46) respectively in low NLR group,but 91.7％ (33/ 36),83.3％ (30/36) and 52.8％ (19/36) respectively in high NLR group (all P＜0.05).Cox regression showed that TNM stage and NLR were independent risk factors for the prognosis of the middle-aged and elderly patients with colorectal cancer (P＜0.05 or P＜0.01).Conclusions Preoperative NLR ＞ 3.13 suggest that the prognosis is poor in middle-aged and elderly patients with colorectal cancer.

Objective To assess the proficiency of water monitoring laboratory at rural, county, and provincial levels in Guangdong province, to ensure the province's drinking water monitoring results accurate and reliable. Methods State quality of certified reference materials as water arsenic, fluoride and chloride of 90 copies each were numbered and distributed to 90 monitoring laboratories in the province for testing, The measurement results of the participatory labs were evaluated through normative values and expanded uncertainty, and were compared with those of robust statistics method. Results All participatory labs had timely feedback of their measurement results. The qualified rate was higher when arsenic was tested by hydride generation atomic fluorescence spectrometry and zinc-new silver salt of sulfuric acid spectrophotometric system, while fluoride and chloride by ion chromatography. The average qualified rates of water arsenic, fluoride and chloride of the province's rural drinking water quality monitoring laboratory were 66% (59/90), 72% (65/90) and 72% (65/90), respectively.Seven participatory labs failed the proficiency testing of all three analytes and unqualified rate was 7.8% (7/90)among the ninety participated monitoring labs. The qualified rates of robust statistics method for arsenic fluoride and chloride were greater than those evaluated by the expanded uncertainty, and large deviations with small sample sizes. Conclusions The testing ability of drink-water monitoring labs in Guangdong province has improved.However, by comparison with the requirements of national quality control and testing skills, there is still a gap. It is suggested that internal quality control be included in routine inspeetion to improve laboratory testing technology.

OBJECTIVETraditional detection approaches for non-O157 STEC are both time and labour consuming in diseases surveillance. Virulence genes detection based on multiplex PCR could not only improve the detection efficiency but also increase the accuracy.

METHODSSix virulence genes of non-O157:H7 (stx1, stx2, eae, hly, etpD, katP6) were detected by two groups of trebling PCRs. The multiplex PCRs were optimized by melting curve analysis in SYBR Green I real-time PCR. Testing result of multiplex PCR was consistent with serological testing.

RESULTSThe sensitivity limits of the multiplex PCR for stx1, stx2, eaeP, etpD, katP, and hly were 10 ng/ml, 120 ng/ml, 110 ng/ml,165 ng/ml, 85 ng/ml, and 15 ng/ml, respectively, which is similar with that of single PCR. When the multiplex PCR was applied in 120 adults and 90 children diarrhea samples detection, 13 cases were detected for non-O157 positive.

CONCLUSIONThe method we established can be used for non-O157 STEC virulence genes detection and screening with high efficiency and accuracy.

Escherichia coli Infections ; diagnosis ; microbiology ; Escherichia coli Proteins ; genetics ; Humans ; Multiplex Polymerase Chain Reaction ; methods ; Shiga-Toxigenic Escherichia coli ; genetics ; isolation & purification ; Virulence Factors ; genetics