1.IFN-? Release Assays for Rapid Diagnosis of Active Tuberculosis:Clinical Application
Hua ZHANG ; Shengwen HUANG ; Zhenyuan LUO
Chinese Journal of Nosocomiology 2009;0(14):-
OBJECTIVE To evaluate the clinical application of T-cell based IFN-? release assays(IGRA) for the rapid diagnosis of active tuberculosis.METHODS IFN-? and HIV antibody were detected by using ELISA.Antibody to Mycobacterium tuberculosis was detected by colloidal gold.At the same time,the M.tuberculosis DNA loads were examined by FQ-PCR.Statistical analysis were performed to analyze the correlation of IFN-? with M.tuberculosis antibody and DNA,respectively.RESULTS The sensitivity of TB-IGRA was 90.24%,specificity was 93.34%;the positive rate of TB-IGRA in 82 tuberculosis patients was higher than from sputum smear(64.63%),TB-PCR(76.83%) and tuberculosis antibody(40.24%).CONCLUSIONS As a replacement of TB-PCR,IFN-? can be used as a valued index to evaluate tuberculosis infectin.
2.Mechanisms of clearance of duck hepatitis B virus from infected adult ducks
Ni TANG ; Ailong HUANG ; Zhenyuan QI ; Al ET
Chinese Journal of Immunology 1985;0(02):-
Objective:To gain insight into the mechanism responsible for clearance of natural hepa DNA virus infections.Methods:A group of seven 2~3 month old ducks were infected intravenously with 10~20 ml DHBV positive serum containing 5?10 7 genomes/ml.Following inoculation,ducks were bled at weekly intervals to obtain serum samples for analysis of DHBV DNA and DHBsAg and anti DHBV antibodies.Peripheral blood mononuclear cells were collected at 10,35 days postinoculation(p.i) and used to conduct antigen specific blastogenesis assay.Liver samples were obtained at 5,30,60 days p.i for analysis of DHBV DNA and surface antigens and liver histology.Results:Infection of all 7 animals with approximately 5?10 8~1?10 9 DHBV genomes led to a transient viremia after an incubation period of 1 to 2 weeks.Liver samples contained multiple copies of all of the expected species of DHBV DNA replicative intermediates,including DHBV cccDNA during the peak of viremic phase.Further analysis showed that the absence of a prolonged viremia could be explained by immediate antigen specific blastogenesis and high titer of antibody response.Meanwhile,there was no obvious evidence of liver cell injury during transient DHBV infection.Conclusion:These results demonstrate that noncytopathic antiviral mechanisms make a role in hepa DNA virus clearance.
3.Extreme lateral lumbar disc herniation after pedicle screw fixation and interbody fusion cage:lumbar stability at long-tem follow-up
Ruiming SHI ; Guosheng LI ; Yifeng ZHANG ; Zhenyuan HUANG ; Li SUN ; Cun WANG
Chinese Journal of Tissue Engineering Research 2014;(40):6464-6470
BACKGROUND:Extreme lateral lumbar disc herniation is a rare type of lumbar disc herniation, there are a variety of treatment methods, but the therapeutic efficacy and recurrence rate are controversial.
OBJECTIVE:To investigate the availability of lumbar pedicle screw fixation combined with interbody fusion cage for treating extreme lateral lumbar disc herniation.
METHODWe retrospectively analyzed 19 patients with extreme lateral lumbar disc herniation after treatment with lumbar pedicle screw fixation combining with interbody fusion cage from March 2006 to January 2009. The outcomes were evaluated depending on VAS scoring standard and Macnab scoring standard, lumbar stability were observed postoperatively. We analyzed the spinal stability in recurrent lumbar disc herniation patients after lumbar pedicle screw fixation combined with interbody fusion cage depending on literature search.
RESULTS AND CONCLUSION:Al the 19 patients were fol owed up for 13 months to 3 years, the leg and lumbar pain of al the patients were relieved to varying degrees. Preoperative VAS score was 7.3±1.28 points and postoperative VAS score was 2.1±0.8 points, showing significant difference between two groups (P<0.05). The excellent and good rate was up to 95%with 15 excellent results, 3 good results and 1 acceptable result depending on Macnab evaluation standard. There was no pedicle screw loosening, broken, non-fusion phenomenon. Al the lumbar interbody fusions were good. No one occurred secondary lumbar spinal stenosis. Experimental findings indicate that, lumbar pedicle screw fixation combined with interbody fusion cage for extremely lateral lumbar disc herniation, is characterized as fast symptom relief, strong fixation and good lumbar stability.
4.Comparison of EEG complexity between rats under awaking and anesthesia
Chunfang GAO ; Genbao ZHANG ; Xiaohua LU ; Xiaoyan XU ; Huixian HU ; Zhenyuan XU ; Lei HUANG
The Journal of Clinical Anesthesiology 2017;33(1):63-65
Objective To compare the EEG complexity between rats under awaking and differ-ent depth of anesthesia via analyzing sample entropy and fractal dimension.Methods Sixteen SD rats were intraperitoneally injected with urethane twice,first with 500 mg/kg and second with 800 mg/kg one hour later.The scalp EEG was collected in stage of awaking (W),light anesthesia (LA)and heavy anesthesia (HA).The sample entropy (SampEn)and fractal dimension (FD)were computed by MATLAB.The characteristic values were denoised by linear dynamic system method during the whole process.Results The value of SampEn and FD gradually dropped from awaking to heavy anes-thesia.The SampEn and FD in W was significantly higher than the value in LA or in HA (P <0.05). The SampEn and FD in HA was significantly lower than that in LA (P < 0.05 ).Conclusion The SampEn and FD of EEG could be used to monitor the depth of anesthesia.
5.Fourier Transform Infrared Microspectroscopy of Rat Kidney with Regard to Fa-tal Hyperthermia
Zhijun WANG ; Shanshan SHEN ; Kaifei DENG ; Zhiqiang QIN ; Ping HUANG ; Zhenyuan WANG
Journal of Forensic Medicine 2015;(4):257-261
Objective T o observe the chem ical groups changing in rat kidney w ith regard to fatal hyper-therm ia by Fourier transform infrared m icrospectroscopy (FT IR-M SP ) and to provide a new m ethod to diagnose fatal hypertherm ia. Methods R ats w ere sacrificed by hypertherm ia, brainstem injury, m assive hem orrhage and asphyxiation and divided into groups. T he renal sam ples w ere dissected im m ediately af-ter death. T he data of infrared spectroscopy in glom erulus w ere m easured by FT IR-M SP. Results T he absorbances of 3 290, 3 070, 2 850, 1 540 and 1 396 cm -1 significantly increased (P<0.05),and the ratios of A1650/A3290 and A1650/A1540 significantly decreased (P<0.05) in group of hypertherm ia. Conclusion FTIR-M SP can analyze the changes of chem ical groups of kidney as an auxiliary diagnosis for discrim inating hyper-therm ia w ith other causes of death.
6.Analysis of microbiological trends and antibiotic susceptibility in 711 episodes of peritoneal dialysis-related peritonitis
Yijing TONG ; Hao YAN ; Zhenyuan LI ; Jiaying HUANG ; Aiping GU ; Zhaohui NI ; Wei FANG
Chinese Journal of Nephrology 2017;33(8):601-608
Objective To investigate the microbiological trends and antibiotic susceptibility of peritoneal dialysis(PD)-related peritonitis (PDAP).Methods All patients who developed PDAP between 2004 and 2015 in Renji Hospital,Shanghai Jiao Tong University School of Medicine were enrolled.Demographic data,results of dialysate pathogen culture and drug susceptibility test were recorded.The trend of peritonitis incidence was measured by Poisson regression and the chi-square test or Fisher exact test method was used to compare the composition of causative organisms and their antimicrobial susceptibilities over time.Results During the study period,a total of 711 episodes of PDAP were occurred in 386 patients.The culture positive rate of pathogens rose from 52.0% in 2004 to 77.0% in 2015 (P < 0.001).The distribution of causative organisms of the culture positive peritonitis was gram-positive bacteria (270,59.5%),followed by gram-negative bacteria (129,28.4%),polymicrobial(39,8.6%),fungi (15,3.3%) and mycobacteria (1,0.2%).From 2004 to 2015,the incidence of peritonitis decreased from 0.214 to 0.160 episodes/patient·year (P=0.034).The incidence of coagulase-negative staphylococcus peritonitis decreased from 0.049 to 0.027 episodes/patient · year (P=0.025),while others had no significant change;A significant decline was observed in the sensitivity of Gram-positive strains to the first generation cephalosporin and ampicillin/sulbactam in 2010-2015 group compared with those in 2001-2009 group (61.3% vs 88.2%,P < 0.001;61.7% vs 85.5%,P=0,001),whereas the sensitivity to vancomycin remained the same.The sensitivity of Gram-negative strains to ceftazidime and amikacin showed no significant change.As for the gram-positive peritonitis treated with cefradine as empirical treatment,compared with those in 2004-2009 group,in 2010-2015group the proportion of patients requiring to change their treatment regime was significantly higher,and the treatment course was longer.Conclusions A gradual decline is observed in the incidence of PDAP and the culture positive rate of pathogens improves.Peritonitis caused by coagulase-negative staphylococcus decreases overtime.The present empirical treatment protocols may need re-evaluation considering the decreased rate of the first generation cephalosporin sensitivity in recent years.
7.The quantitative analysis of S100 in the brain tissue and serum following diffuse brain injury in rats
Qi WANG ; Ping HUANG ; Bo XING ; Ya TUO ; Yongpan ZHANG ; Weiping TIAN ; Zhenyuan WANG
Journal of Pharmaceutical Analysis 2007;19(1):118-封3
Objective To investigate the dynamics of the level of S100 in cerebrum, brainstem, and serum following the diffuse brain injury in rats and provide the experimental evidences for estimating injury time. Methods ELISA was used to determine whether S100 protein is changed after diffuse brain injury in rats. Forty rats were sacrificed at 0.5 hour, 2 hours, 4 hours, 12 hours, 24 hours, 3 d and 7 d after diffuse brain injury and normal rats as control. Results The level of S100 in cerebrum, brainstem, and serum increased, followed by a decrease, and then further increased. The level of S100 could be detected to increase at 30 minutes and reached the peak at 4 hours after DBI. The level decreased gradually to the normal at 1d and till 3 d formed the second peak. The level returned to the normal at 7d following injury again. In the postmortem injury groups, there were no significant changes compared to the control group. Conclusion The present study showed that the time-dependent expression of S100 is obvious following diffuse brain injury in rats and suggested that S100 will be a suitable marker for diffuse brain injury age determination.
8.Establishment and evaluation of the method for detecting HBV DNA in serum using HBV DNA probe labeled directly by alkaline phosphatase.
Yaxi CHEN ; Ailong HUANG ; Zhenyuan QI ; Youlan SHAN ; Hang SUN
Chinese Journal of Hepatology 2002;10(6):429-431
OBJECTIVETo establish a sensitive and specific technique for detecting HBV DNA in serum using HBV DNA probe labeled directly by alkaline phosphatase (AlkPhos Direc probe).
METHODSThe probe that purified HBV DNA sequence was labeled directly by alkaline phosphatase and chemiluminescent substrate CDP-star for AP was used in the hybridization assay. HBV DNA was detected by autoradiography on the film. The test compared the chemiluminescen dot blot hybridization assay for 80 samples with digoxigenin-labeled HBV DNA probe detective method. The correlation of 70 samples test results between fluorescent quantitative HBV DNA PCR method and dot blot hybridization assay by AlkPhos Direc probe was analysed.
RESULTSThe sensitivity of the probe labeled directly by alkaline phosphatase was 10pg at least. The coincidence was 100% compared with digoxigenin-labeled HBV DNA probe detection. A correlation coefficient of HBV DNA quantitative results between fluorescent quantitative HBV DNA PCR (QPCR) method and dot blot hybridization assay by AlkPhos Direc probe was 0.98 (P<0.01).
CONCLUSIONSThe method detecting HBV DNA in serum by HBV DNA AlkPhos Direc probe is sensitive and specific. The results between two methods with AlkPhos Direc and digoxigenin-labeled HBV DNA probe are coincident completely. The correlation of HBV DNA quantitative results between fluorescent QPCR method and dot blot hybridization assay by AlkPhos Direc probe is satisfactory.
Alkaline Phosphatase ; chemistry ; metabolism ; Animals ; DNA Probes ; chemistry ; genetics ; DNA, Viral ; blood ; genetics ; Hepatitis B ; blood ; diagnosis ; virology ; Hepatitis B virus ; genetics ; Humans ; Molecular Diagnostic Techniques ; methods ; standards ; Polymerase Chain Reaction ; methods ; Sensitivity and Specificity
9.Metabolism of mangiferin by human intestinal bacteria in vitro.
Huixue HUANG ; Zhenyuan TAN ; Jiagang DENG ; Qiuyun LIANG ; Yumei NONG ; Nianmei SONG
China Journal of Chinese Materia Medica 2011;36(4):443-445
OBJECTIVETo study the metabolism of mangiferin by human intestinal bacteria in vitro.
METHODHuman intestinal bacteria and mangiferin were incubated under anaerobic conditions in vitro. The metabolite was separated and purified by D101 macroporous resin column and preparation high performance liquid chromatography, and its structure was identified by MS and NMR.
RESULTAfter 12 h incubation with human intestinal bacteria, the content of mangiferin metabolite reached the maximum, and it was determined as 1, 3, 6, 7-tetrahydroxyxanthen by MS and NMR.
CONCLUSIONMangiferin can be metabolized in vitro by human intestinal bacteria into its aglycone (1, 3, 6, 7-tetrahydroxyxanthen).
Bacteria ; metabolism ; Chromatography, High Pressure Liquid ; Humans ; Intestines ; microbiology ; Xanthones ; metabolism
10.Application of the pyrosequencing technique for detection of VKORC1 and CYP2C9 genotypes
Qin XU ; Shengwen HUANG ; Nannan YANG ; Shimin WANG ; Zhenyuan LUO ; Bangquan AN
International Journal of Laboratory Medicine 2018;39(7):769-772
Objective To establish genotyping methods for vitamin K epoxide reductase complex subunit 1 (VKORC1)and cytochrome P450 2C9(CYP2C9)based on pyrosequencing technique to detection of warfarin metabolizing enzyme related gene polymorphisms.Methods A total of 50 peripheral blood samples from healthy adults were collected and the whole blood genomic DNA was extracted.A set of biotin-labeled amplifi-cation primers and sequencing primers were designed respectively for three SNP sites:VKORC1 -1639 G>A,CYP2C9 430C> T and CYP2C9 1075A>C.After PCR amplification of the samples,pyrophosphoric acid se-quencing was conducted.And then the signal peaks form were combined to analyze and determine each sample genotype.Genotyping results were verified by Sanger sequencing,and the consistency of the two sequencing methods was compared.Results Genotypes of the three SNPs can be clearly determined according to the ba-ses and height of the signal peaks.Among the 50 samples,there were 41 AA and nine AG for VKORC1 -1639G>A,accounting for 82% and 12% respectively,and there were 45 *1/*1,five *1/*3 for CYP2C9, accounting for 90% and 10% respectively,no CYP2C9*2 allele detected.Genotype results detected by pyrose-quencing and Sanger sequencing were consistent with each other.Conclusion In SNP genotyping,Pyrose-quencing has the advantages of convenience,time-saving,cheap with accurate and reliable results,which can quickly determine the genotypes of CYP2C9 and VKORC1.