1.Progress and challenges in the laboratory diagnosis of hepatitis C virus
Chinese Journal of Laboratory Medicine 2008;31(8):845-848
Hepatitis C virus (HCV) is a common blood-borne pathogen that relies heavily on laboratory assays for the confirmation of infection. Anti-HCV testing is the earliest and classical method which is easy to perform but has a long window period (7 - 8 weeks with the third-generation method) compared to the nucleic acid test (NAT). NAT provides direct evidence for the presence of HCV and quantitative HCV RNA testing has been applied to monitor the antiviral response to treatment. In some developed countries NAT is also used for blood screening. Cost-effectiveness and standardization are the major challenges for NAT. In recent years HCV core antigen assay and the combination antigen-antibody assay have been introduced in some clinical laboratories and proved to be premising in the early diagnosis of HCV, whereas they still remain less sensitive than NAT and require methodological improvement. Finally, HCV genotyping assays based on sequencing or reverse hybridization can provide important prognostic information related to therapeutic response, however, it is rarely used in China due to the high cost.
2.Resistance mutation during anti-HBV and anti-HCV treatment:question and strategy
Chinese Journal of Laboratory Medicine 2013;(3):193-200
Both long term follow up and real world research on chronic hepatitis B have accumulated data on nucleos (t) ide analogues in anti-HBV treatment,which shows that resistance mutation turns out to be the major obstacle in achieving response.Identification of genotypic resistance at early stage is key to improve strategy.Deep-sequencing will be helpful to predict resistance earlier.Direct acting autiviral agents on hepatitic C virus tell us resistance at the very beginning,however,more data is still needed to elucidate how to use resistance monitoring of anti-HCV treatment.
3.EFFECT OF HYPERTHYROIDISM ON DEVELOPMENT OF ENCEPHALIC CHOLINER GIC NEURONS DURING THE CRITICAL STAGE OF BRAIN DEVELOPMENT IN RATS
Zhaoying LI ; Ruifeng YANG ; Zupei CHEN
Chinese Journal of Endocrinology and Metabolism 1986;0(03):-
The choline acetyltransferase (CHAT) and the molecular forms of acetyl-cholinesterase (ACHE) activites in various brain regions of 20-day-old hypothyroid and hyper-thyroid rats were measured. The results provided the following information: 1) CHAT and ACHE activities were directly interrelated with thyroid hormones. 2) In both hypothyroid and hyperthyroid rats the nonextractable ACHE activity was distinctly decreased in every brain region, suggesting that both conditions were affected in the critical period of cholinergic synaptic development. 3) The ratio of membrane-bound ACHE to soluble ACHE decreased;it showed that thyroid hormone deficiency might distrub development and maturation of cholinergic neurons. 4) In most regions of the central nervous system,the CHAT seemed to be more affected than ACHE by thyroid hormones.
4.Dose Determination and Calibration of Siemens primus E LINAC
Ruifeng YANG ; Hui HUANG ; Tao HE
Chinese Medical Equipment Journal 1989;0(04):-
The procedures for the calculation and measurement and calibration of output dose of the Siemens primus E accelerator with IAEA regulation are introduced to probe an accurate,rapid and effective method.The parameter value of measurement is determined according to IAEA regulations and absorbed dose calculation formal is briefed to the form that the reading of instrument multiplies exposure calibration coefficient Nx and Air temperature,pressure and humidity effects Ktp and cumulative correction factor Cf the precision of accelerator conform to the standard of QA&QC by the measurement and calibration of the Hospital Primus E accelerator in output dose.The methods of measurement and calibration can be used for the determination of output dose in precise radiotherapy of digital medical linear accelerator and clinical treatment.
5.Study of Arsenic Trioxide and Cisplatin on Human Ovarian Cancer Cell.
Guohong YANG ; Hua JIN ; Ruifeng LIU
Journal of Medical Research 2006;0(02):-
2?mol/L) was obvious and the inhibitive degree was nearly 100% when the concentrations of arsenic trioxide and cisplatin were 12?mol/L and 4?mol/L. 2.Higher concentrations of arsenic trioxide and cisplatin perturbed cell through S-phase and up-regulated p53 gene expressions.Conclusions The inhibitive effect of arsenic trioxide combined with cisplatin was more obvious than either one used separately; the apoptosis mechanism of ovarian cancer cell line HO-8910 is related with perturbing of S phase and up-regulating of p53 gene expression.
6.HCV genotyping and host genotyping: what role will they play in the antiviral treatment
Jian WANG ; Ruifeng YANG ; Lai WEI
Chinese Journal of Laboratory Medicine 2012;35(2):97-100
Persistent infection of hepatitis C virus (HCV) remains as a worldwide threat to public health,which involves a complex interaction between virus- and host related factors.HCV is classified as six genotypes and many subtypes according to the sequence heterogenecity.HCV genotype should be determined prior to treatment initiation since it plays a key role in selection of therapeutic regimen for chronic hepatitis C.Development of the antiviral treatment with protease inhibitor in combination with pegylated IFN-α and ribavirin requires the accurate determination of subtypes,e.g. 1a and 1b,as well.Genotyping methods based onsequenceanalysis, reversehybridizationorreal-timePCRhavebeendevelopedand evaluated.Some issues,however,should be settled to standardize the utility and result interpretation of these methods.More recently,host genotypes of IL28B have been found to be closely associated with HCV spontaneous clearance and the response to antiviral therapy.Moreover,polymorphisms in inosine triphosphate pyrophosphatase gene affect ribavirin-induced anemia.Therefore, host genotyping will be beneficial in predicting the outcome of chronic hepatitis C and monitoring the drug-induced adverse events.
7.The effect of therapeutic life style on serum FFA and constitution index in patients with coronary heart disease complicated metabolic syndrome
Xiaoyong LIU ; Ge ZHANG ; Ruifeng YANG ; Zhi LIN ; Rui YANG
Chongqing Medicine 2015;(22):3099-3101,3104
Objective To investigate the level of serum free fatty acid (FFA )after improving the life style in patients with coronary heart disease complicated metabolic syndrome and the effect of therapeutic life style on traditional risk factors of coronary artery disease.Methods A total of 395 patients with coronary heart disease complicated metabolic syndrome were recruited.Pa-tients were divided into intervention group (group A,conventional drug therapy+ intensive life style intervention,n=97)and non-intervention group (group B,conventional drug therapy,n=38)according to the scores of life style.Serum free fatty acid (FFA) was determined by ELASA.The scores of life style was obtained bylife style questionnaire.Results (1)The serum FFA of pa-tients with coronary heart disease complicated metabolic syndrome were positively related to waist circumference and waist-high-ra-tio.(2)Waist circumference,BMI and FFA of group A were significantly lower than those in group B after therapeutic life style in-tervention(P <0.05).(3)Compared with the baseline,the constitution index and FFA in group A were significantly lower after 6-months therapeutic life style intervention(P <0.05).Conclusion Therapeutic life style can reduce the level of FFA and constitution index of the patients with coronary heart disease complicated metabolic syndrome.
8.Effects of bone marrow mesenchymal stem cell transplantation on CD4+CD25+regulatory T cells in rats with primary nephrotic syndrome
Huandan YANG ; Ruifeng ZHANG ; Dongjin FENG ; Bingbing ZHU ; Juan LV
Chinese Journal of Tissue Engineering Research 2014;(1):33-38
BACKGROUND:Decreased function and reduced number of CD4+CD25+regulatory T cells have been considered the major manifestation of immunity dysfunction in children with primary nephrotic syndrome. Bone marrow mesenchymal stem cells have immunoregulation effects, which up-regulate CD4+CD25+regulatory T cells, inhibit proliferation of lymphocytes, and have been widely used in many immune diseases.
OBJECTIVE:To investigate the effects of bone marrow mesenchymal stem celltransplantation on the CD4+CD25+regulatory T cells of peripheral blood in rats with primary nephrotic syndrome.
METHODS:Bone marrow mesenchymal stem cells from six Sprague-Dawley rats were isolated, passaged and utilized for cellsuspension preparation. At the third passage, bone marrow mesenchymal stem cells were used for transplantation. The remaining 30 rats were randomly and equal y divided into three groups:normal group, normal saline infusion group, and bone marrow mesenchymal stem cells group. The rat models of primary nephrotic syndrome were established by single injection of adriamycin intravenously through tail vein in the latter two groups. Rats were then treated with bone marrow mesenchymal stem cells (1×10 7 ) (bone marrow mesenchymal stem cells group) or normal saline (normal saline infusion group) through tail vein at the same time after adriamycin administration. The normal group received no treatment.
RESULTS AND CONCLUSION:Compared with the normal group, rats in the normal saline infusion group developed nephropathy characterized by ascites, proteinuria, hypoalbuminemia, hypercholastero-lnemia, and progressive renal injury. However, the proteinurine and clinical severity in bone marrow mesenchymal stem cells group were significantly ameliorated after treatment with bone marrow mesenchymal stem cells. CD4+CD25+Treg/CD4+Treg in the peripheral blood in the bone marrow mesenchymal stem cells group and normal saline infusion group were significantly higher than that in the normal group at 28 days after model establishment (P<0.05), while there was no significant difference between bone marrow mesenchymal stem cells group and normal saline infusion group (P>0.05). The expression of FoxP3 mRNA in the peripheral blood mononuclear cells of the bone marrow mesenchymal stem cells group was significantly higher than that in the normal saline infusion group and normal group (P<0.05). The bone marrow mesenchymal stem cells play a protective effect in rats with primary nephrotic syndrome, which may be related to the increase of local expression of FoxP3 and generation of CD4+CD25+Treg.
9.A primary study on the level of human cytomegalovirus specmc CD8+ T lymphocytes and IFN-γ secretion effect in kidney transplant recipients
Ruifeng YANG ; Guobin XU ; Xu WU ; Zhiyan LI ; Tiean XIA
Chinese Journal of Laboratory Medicine 2008;31(9):1016-1020
Objective To investigate the level of human cytomegalovirus(HCMV)specific CD8+ T lymphoeytes in peripheral blood and the immune reaponae of HCMV reactivation after kidney transplantation.Methods Thirty-eight HCMV seropesitive HLA-A*0201 kidney transplant recipients(9 with HCMV infection and 29 without HCMV infection)and 54 healthy individuals were enrolled.The levels of total HCMV specific CD8+ T cells were measured using HLA-A2 pentamer folded with HCMV-peptide NLVPMVATV.The levels of IFN-γ secreting CD8+ T cells were measured by intracelluhr IEN-γ staining pulsed with the same peptide.Results The median levels of pentamer stained CD8+ T cells were 1.19%(0-19.42%),1.20%(0-18.40%)and 3.2%(0.51%-18.90%)in healthy group,negative HCMV group and positive HCMV group(H=5.34,P>0.05),respectively.The median levels of IFN-γ secreting CD8+ T cells were 0.72%(0-0.70%),0.47%(0-5.61%)and 0.67%(0.07%-4.00%),respectively(H=0.58,P>0.05).However,the mean proportions of IFN-γ secreting pentamer stained T cells relative to total HCMV specifc CIL were(60.18±19.16)%,(39.19±17.22)% and(20.02±13.26)%,respectively.There were significant differences among the groups(P<0.01).Condusiorm There was no significant difference of levels of HCMV specific CD8+ T lymphocytes in peripheral blood between the kidney transplant recipients and healthy individuals.However,the proportion of HCMV-specific IFN-γ producing CD8+ T cells in pentamer stained cells was reduced in the kidney transplant recipients especially in those with active HCMV infection,which may contribute to the inability to control HCMV reactivation.
10.Evaluation of the performance of a DNA microarray method for detecting HBV mutations conferring antiviral resistance to lamivudine and adefovir
Ruifeng YANG ; Shaocai DU ; Xu CONG ; Hui MA ; Lai WEI
Chinese Journal of Laboratory Medicine 2010;33(11):1049-1053
Objective To evaluate the performance of a DNA microarray method for detecting HBV antiviral drug-resistant mutations. Methods Two hundred and twenty four serum samples from patients with CHB were tested in parallel by DNA microarray and direct sequencing for the mutations within the HBV reverse transcriptase (rt) region, which included rtL180, rtA181, rtM204 and rtN236. Samples with discrepant results were retested by clonal sequencing. Results Complete concordance between DNA microarray and direct sequencing results was observed in 214 out of 224 samples (95. 5% ). The presence of mixed viral populations in the other 10 samples detected by DNA microarray but not by direct sequencing was confirmed by clonal analysis. The DNA microarray could detect minor viral populations which constituted 5.0%-15. 0% of the total viral load. Conclusion DNA microarray is highly consistent with direct sequencing in detecting HBV mutations conferring drug resistance and more sensitive in detecting mixed mutant and wild-type sequences than direct sequencing, which makes it a useful tool for early detection of drug resistance early.