Objective To report the HLA data of volunteer donors of Chinese bank from Northwest China and characterize the distribution of HLA genes in Northwest China. Methods HLA-A, B antigens of 2315 volunteer donors were examined by the method of microlymphocytetoxicity (MLT) test .The antigen frequencies(AF) were assessed by directly counting; and based on that gene frequencies(GF) were calculated. HLA data from other population were collected and distribution characteristics were compared. With the raw data, Hardy-Weinberg equilibrium, statistical parameters of forensic medicine interest for HLA were computed. Results A total of 18 specific antigens were detected in HLA-A and the most frequent antigen was A2 . AF and GF were 0.5136 and 0.3026, respectively. A total of 42 specific antigens were detected in HLA-B and the most frequent antigen was A13. Its AF and GF were 0.1978 and 0.1044, respectively. The heterozygosity(H), polymorphism information content(PIC), discrimination power(DP) and probability of paternity exclusion (PPE) of HLA-A were 0.8215, 0.8212, 0.9356 and 0.7798 accordingly; while H,PIC, DP and PPE of HLA-B were 0.9322, 0.9322, 0.9878 and 0.9528. Conclusion The polymorphism of HLA-A,B genes is characteristic in Chinese. In this research, the genetic trait of HLA in 2315 volunteers is consistent with Northern Han population.

OBJECTIVESTo investigate the profile of liver histological damage after autologous adoptive immunotherapy with cytokine-induced killer cells (CIK cells) in patients with chronic hepatitis B (CHB).

METHODS16 CHB patients were randomly enrolled and received autologous adoptive immunotherapy, then followed up 52 weeks. Liver samples were taken from the patients to evaluate the degree of inflammation and fibrosis. The markers of hepatitis B virus and liver function were also detected.

RESULTS4 patients had two liver-biopsied samples before therapy and after 52 weeks follow-up. One patient's histological assessment revealed a significant improvement in intralobular necroinflammation (G2 --> G1) and fibrosis (S2 --> S1). The others failed to show obvious changes in liver histology. After 10 days culture in vitro, phenotypic characterization of CIK cells changed significantly by flow cytometry. The percentage of CD4+ cells decreased gradually, while the percentage of CD8+ cells increased from 20% to 60% - 80%. After 52 weeks follow-up, HBV DNA was negative (HBV DNA<4pg/ml in serum) in 6 out of 14 patients. The rates of both HBeAg/anti-HBe seroconversion and alanine aminotransferase normalization were 42.86%(6/14). There was no HBsAg/anti-HBs seroconversion. There was few severe treatment-related adverse events.

CONCLUSIONAutologous adoptive immunotherapy doesn't induce the damage of liver histology in chronic hepatitis B patients, which inhibits hepatitis B virus replication by a certain noncytotoxic mechanism.

Adolescent ; Adult ; Aged ; CD4-CD8 Ratio ; DNA, Viral ; analysis ; Hepatitis B, Chronic ; immunology ; pathology ; therapy ; Humans ; Immunotherapy, Adoptive ; adverse effects ; Liver ; pathology ; Middle Aged

OBJECTIVEThe present aimed to observe the effect of phosphatase inhibitor cyclosporine A on the subcellular location and on expression of HBcAg in HepG2.2.15 cells.

METHODSThirty micrograms/ml of cyclosporine A (CSA) was added into HepG2.2.15 cell culture system and on days 2 and 4 HBcAg and HBsAg were respectively stained with fluorescent immunocytochemistry and observed under confocal microscope. Cell apoptosis was detected by terminal deoxynucleotidyl transferase-mediated dUTP nick end labelling method.

RESULTSHBcAg was mostly expressed in cytoplasm in the control HepG2.2.15 cells. After 2 days CSA administration of the expression of HBcAg and HBsAg in cytoplasm significantly decreased and the signals of HBcAg in nucleus increased , whereas the HBcAg was still mainly expressed in nucleus in about 1/4 of the cells. Cell apoptosis was observed in about 30% of the cells.

CONCLUSIONCSA improves the nuclear entry of core protein. The increase of HBcAg in nucleus was likely to be related with it's phosphorylation and cell aging or apoptosis.

Active Transport, Cell Nucleus ; Apoptosis ; Cell Line, Tumor ; Cell Nucleus ; metabolism ; Cyclosporine ; pharmacology ; Cytoplasm ; metabolism ; Hepatitis B Core Antigens ; analysis ; metabolism ; Hepatitis B Surface Antigens ; analysis ; Humans ; In Situ Nick-End Labeling ; Phosphorylation

OBJECTIVESThe hepatitis B virus core protein has been found in nuclei, cytoplasm, or both of hepatocytes transfected with HBV DNA. It is still unclear whether intact core particles could pass through nuclear pores and what could be the mechanism regulating the subcellular localization of the core protein. This study on the distribution of core protein in hepatocytes and its translocation has a potential advantage to learn more about the HBV life cycle.

METHODSDimethyl sulphoxide (DMSO, 2%), which effects hepatic differentiation, and/or 1 micro mol/L heteroaryldihydropyrimidine Bay41-4109, which interferes with the assembly of core particles, were added into HepG2.2.15 cell culture system for 4 days. The hepatitis B virus core antigen (HBcAg) and hepatitis B virus surface antigen (HBsAg) were stained with fluorescent immunocytochemistry and then observed under a confocal microscope. HBcAg in cytoplasm and nuclei were respectively extracted and analyzed using Western blot. HBV covalently closed circular DNA (cccDNA) was detected by using selective PCR method.

RESULTSThe HBcAg was mostly expressed in the cytoplasm and weak signals of cccDNA were detected in the control HepG2.2.15 cells. After DMSO treatment, the expression of HBcAg in cytoplasm was increased about 2.5-fold; the expression of HBcAg and cccDNA in nuclei also increased. With the use of Bay41-4109, the signal of HBcAg in cytoplasm decreased 2/3, but it increased in the nuclei, and cccDNA decreased in the nuclei. When the HepG2.2.15 cells were treated both with DMSO and Bay41-4109, cord-liked distribution of HBsAg was observed in the cytoplasm. HBcAg in cytoplasm was decreased 1/2 but the HBcAg in the nuclei increased about 5-fold, whereas the cccDNA was almost negative.

CONCLUSIONIn HepG2.2.15 cells, the core protein is mainly assembled as a formation of core particles in the cytoplasm and they are blocked by the nuclear membrane. Bay41-4109 interferes with the assembly of core particles and the dissociated core proteins are able to enter the nuclei. DMSO promotes the nuclear entry of core protein/core particles and facilitates the formation of cccDNA.

Chromosome Positioning ; Dimethyl Sulfoxide ; pharmacology ; Hep G2 Cells ; Hepatitis B Core Antigens ; metabolism ; Hepatitis B virus ; physiology ; Humans ; Neoplasm Metastasis ; Pyridines ; pharmacology ; Pyrimidines ; pharmacology ; Viral Core Proteins ; metabolism ; Virus Assembly

AIM:To observe the effects of transection of right cervical sympathetic trunk(TCST)on inflam-matory response and expression of high mobility group box 1(HMGB1)and TLR4/NF-κB signaling pathway in the rats af-ter acute myocardial infarction(AMI).METHODS: AMI model was established by ligation of left anterior descending coronary artery in SD rats,then the model rats were randomly divided into MI group and MI +TCST group.MI+TCST model was performed by transection of right cervical sympathetic trunk after left anterior descending coronary artery ligation. The rats in MI group and MI+TCST group were divided into 1,3,7,14 and 28 d subgroups,and another sham operation group threading without ligation, with 8 rats in above each group.After modeling for 4 weeks, the cardiac function was measured by echocardiography.All rats were killed to harvest the hearts for mesuring cardiac hypertrophy index.The myo-cardial tissue close to infarction was observed with HE staining.The relative mRNA expression levels of HMGB1, tumor necrosis factor α(TNF-α)and interleukin(IL)-6 at different time points were detected by real-time PCR.The protein ex-pression of HMGB1 and TLR4 at different time points after AMI was determined by Western blot.The effect of transection of right cervical sympathetic trunk on the expressions of HMGB 1 and TLR4/NF-κB signaling pathway was also analyzed. RESULTS:Compared with the MI group,left ventricular ejection fraction(LVEF)and left ventricular shorterning fraction (LVFS)were significantly higher(P<0.05),left ventricular end-diastole dimension(LVEDd),left ventricular end-sys-tole dimension(LVESd)and cardiac hypertrophy index were significantly lower(P<0.05), and the mRNA levels of HMGB1,TNF-αand IL-6 decreased significantly in MI +TCST group(P<0.05).Western blot results revealed that the protein expression level of HMGB1 increased in the infarct border zone at 3 d,and reached its peak at 7 d,then gradually decreased,and at 28 d after MI in MI group was still significantly higher than that in sham group(P<0.05).The protein expression of TLR4 was consistent with that of HMGB1.Transection of right cervical sympathetic trunk reduced protein ex-pression of HMGB1 and TLR4/NF-κB signaling pathway-related proteins(P<0.05).CONCLUSION: Transection of right cervical sympathetic trunk improves ventricular remodeling and maintaining cardiac function.The mechanism may be related to inhibiting HMGB1/TLR4/NF-κB signaling pathway to reduce inflammatory response.

OBJECTIVETo set up the drug lymphocyte stimulation test (DLST), as a diagnosis means for DILI which was immunity idiosyncrasy, improve the Diagnosis, level of DILI.

METHODFor the 59 patients who diagnosed as DILI, we separated their PBMC, exploring to the suspicious drug which caused DILI, then use the methods 3H-TdR to test, according to the mixed degree to clear the PBMC count which specific activated by drug.We also set up drug group, negative control and Positive control at the same time. Preliminary experiments was including the best dose of PHA and the best concentration of the drug. We set up 40 healthy group in our experiments as a control, and explore them on the same drug every time. We test the two groups at the same time. We handled the results use t-test.

RESULTSThe methods 3H-TdR could be exactly reflect the PBMC's proliferation degree nearly the same when they were be stimulation by PHA or the sensitive drug. When the DILI patients were explore to the suspicious drug, their stimulation index (SI) Obviously higher than 1.8. Form this test, there were 28 in 59 patients of DILI's group were positive (47.46%), SI was from 1.9 to 43.08, the average was 22.49, the healthy group SI was lower than 1.8, the SI of DILI's group was significantly higher than healthy group (5.78+/-0.75/1.16+/-0.25, P less than 0.05). Our test suggested DLST has Higher specificity (94.92%) and sensitivity (47.46%).

CONCLUSIONDLST was significance for the patients who diagnosed as immunity idiosyncrasy's DILI, it's reflected these patients' Proliferation of PBMC when explored to the suspicious drug for the second time.

Adolescent ; Adult ; Case-Control Studies ; Chemical and Drug Induced Liver Injury ; diagnosis ; Female ; Humans ; Leukocytes, Mononuclear ; Lymphocyte Activation ; Male ; Middle Aged ; Young Adult

OBJECTIVETo explore the effect of small interfering RNA(siRNA) on silence of mdr1 gene and reversal of apoptosis resistance in multidrug-resistant (MDR) human leukemia K562/ADM cell.

METHODSHuman MDR leukemia cell line K562/ADM was used as the target cells. Two siRNAs (mdr1 siRNA-1 and mdr1 siRNA-2) targeted mdr1 gene were chemically synthesized and transfected into K562/ADM cells with liposome. Expression of mdr1 mRNA was determined by real-time PCR, P-glycoprotein (P-gp) expression and caspase-3 activity were measured with flow cytometry (FCM), and the cell apoptosis was observed by optical and electronic microscopy for morphology and Annexin V/PI staining.

RESULTSThe mdr1 siRNA-1 and mdr1 siRNA-2 could markedly down-regulate the expression of mdr1 gene in K562/ADM cells, the expression of mdr1 mRNA decreased by 91.2% and 82.0% , and the P-gp by 74.1% and 84.4%, respectively. The caspase-3 activity was markedly enhanced, and the active caspase-3 in K562/ADM cells increased by about 40% compared to liposome alone and non-silencing controls. the sensitivity of K562/ADM cells to adriamycin-induced apoptosis was significantly augmented, the apoptotic rate of the cells treated with siRNA plus adriamycin increased by about 60% compared to adriamycin alone.

CONCLUSIONsiRNAs silence the expression of mdr1/P-gp to overcome the P-gp-mediated apoptosis resistance in drug-resistant K562/ADM cells.

ATP Binding Cassette Transporter, Sub-Family B ; ATP-Binding Cassette, Sub-Family B, Member 1 ; genetics ; metabolism ; Apoptosis ; Caspase 3 ; metabolism ; Drug Resistance, Multiple ; genetics ; Drug Resistance, Neoplasm ; genetics ; Humans ; K562 Cells ; RNA Interference ; RNA, Messenger ; genetics ; RNA, Small Interfering ; genetics ; Transfection

OBJECTIVETo explore relevant between human cytomegalovirus (HCMV) infection and college students' neurobehaviors.

METHODS87 college students were enlisted. They were tested with Bole. Neurobehavioral evaluation system (B. NES), and HCMV IgG antibody was detected after separation of serum. We analyzed the test results of B. NES by SPSS software.

RESULTS76 college students were infected by HCMV in the past and 11 college students were not infected. The infected group scored 8.89 +/- 6.60 in depression aspect of emotion state test, while control group got 15.73 +/- 9.00. There was Significant difference between infection group and control (P < 0.05). There were no significant differences in other aspects of emotion states, study and memory, perception and mental movement (P > 0.05).

CONCLUSIONHCMV infection is associated with depression status.

Adult ; Cytomegalovirus Infections ; psychology ; Emotions ; Female ; Humans ; Learning ; Male ; Memory ; Students ; psychology ; Universities

OBJECTIVETo study the short teadem repeat(STR) genetics structure of a Chinese Yunnan Yi racial group.

METHODSGenetic distributions for nine STR loci were determined based on STR gene scan marked by fluorescence.

RESULTSSixty-nine alleles and 164 kinds of genotypes were detected and identified from 84 unrelated Yi racial individuals. The corresponding gene and genotype frequencies were in 0.0060-0.5060 or 0.0119-0.4167 respectively. The expected and observed genotype frequencies of nine STR loci were in accordance with the Hardy-Weinberg equilibrium(P>0.05). The statistical analyses of nine STR loci showed that PIC was distributed in 0.5804-0.8777, H was in 0.6507-0.8002, DP was in 0.7976-0.9558, EPP was in 0.5207-0.8386, except TPOX and THO1 loci.

CONCLUSIONAbove research data enrich the Chinese genetic database, and play an important role in Chinese genetic study and in forensic application.

Asian Continental Ancestry Group ; genetics ; China ; ethnology ; Chromosome Mapping ; Ethnic Groups ; genetics ; Genetics, Population ; Humans ; Microsatellite Repeats ; genetics ; Polymorphism, Genetic ; Tandem Repeat Sequences

OBJECTIVETo define TB control priorities using cost-effectiveness and burden of disease.

METHODSAn assumed cohort of 2,000 cases was set up based on age-specific incidence of 794 newly registered smear-positive cases in Beijing in 1994. Prognostic trees and model diagrams of infectivity with natural history and DOTS intervention were constructed based on the epidemiological parameters.

RESULTSDOTS reduced 89.19% of YLL, 78.90% of YLD, and 99.98% of infectivity BOD. One DALY could be saved with 45.70 Yuan by DOTS with 3% discount. Sensitivity analysis showed that discount had effect on CER. Weight of age was insensitive to CER. The higher the DOTS cured rate, the more the cost-effectiveness.

CONCLUSIONSDOTS is a good cost-effectiveness TB control strategy. Cost-effectiveness and burden of disease can be used to define TB control priorities.

Adolescent ; Adult ; Child ; Child, Preschool ; China ; epidemiology ; Cohort Studies ; Cost of Illness ; Cost-Benefit Analysis ; Disabled Persons ; Forecasting ; Humans ; Middle Aged ; Models, Biological ; Quality-Adjusted Life Years ; Tuberculosis ; economics ; epidemiology ; prevention & control