Objective To develop a convenient method efficiently expands the frequency of specific CTLS.Methods We used different concentrations of CMV-speeific epitope peptides pp65 to stimulate PBMCs for expansion of CMV-specific CTLs.CMV-specifie CTLs were doubly labeled by tetramers-PE and CD_8-FITC for FACS analysis.Results The method expands CMV-speeific CTLs efficiently.CMV-specific CTLs were expanded from 1% to 20% of PBMCs quickly(namely 40% of CD_8~+ T cells).The method provided a large number of cells with tetramer staining of CD_8~+ T cells for FACS analysis from a single blood sampling.Conclusions Peptides stimulation methods are convenient,easy to operate and expanded CMV- specific CTLs efficiently.The increased frequencies of CMV-specific CTLs allowed the data of different individuals to be easily compared and sequentially evaluated.The methods lay the base for adoptive immunotherapy to prevent CMV disease.

AIM: To investigate the ocular complication after radiation therapy for nasopharyngeal carcinoma(NPC).METHODS: The authors performed a previous study on keratopathy in 213 NPC patients who received first stage radiation and had at least 10 months of follow-up. These patients were categorized into three groups depending on NPC clinic stages. Rates and proportions of keratopathy occurring in these groups were compared and analyzed with Chi-square Test and Spearman rank correlation coefficient.RESULTS: Radiation keratopathy developed in 19 patients, about 8.9% (19/213). The latency value was 3 to 30days. The effect of NPC clinic stages and radiation did on the development of keratopathy was not statistically significant (P＞0.05).CONCLUSION: The NPC clinic stages and radiation doses plays few effects on the development of keratopathy. It may play a key role that corneal nerves damage induced ocular surface diseases. It can not be excluded that individuals have different sensitivities to radiation.

The relationship of HLA-A, -Cw alleles on HIV infection and AIDS disease progression in the Chinese Yi ethnic group of Sichuan province were investigated. The genetic polymorphisms of HLA-A, -Cw alleles of 102 unrelated healthy Chinese Yi ethnic individuals, 68 HIV-1 infected and 21 HIV positive long-time survivors were typed by PCR-SSP assay. Statistic signifiance was determined by the χ2 test with the SPSS software. No significant differences were observed between the HLA-A, -Cw alleles of the 68 HIV-1 infected and 102 non-infected Chinese Yi control individuals. Whereas the prevalence of A*3601,Cw*14(01-03)and Cw*0304 was significantly higher in 21 long time survivors compared with 102 healthy controls with P values of 0.016, 0.016 and 0.000 by χ2 or the Fisher exact test respectively. The result implies that A*3601,Cw*14(01-03) and Cw*0304 may be associated with slow AIDS disease progression in the Chinese Yi ethnic group, further studies on this association may yield insight on the pathogenesis of HIV-1 infection.

OBJECTIVETo analyze the drift of the complementarity-determining region 3 (CDR3) of T cell receptor (TCR) beta chain variable region (TCR BV) in peripheral blood mononuclear cells (PBMCs) of patients with systemic lupus erythematosus.

METHODSImmunoscope spectratyping techniques was used to analyze the distribution of TCRbeta chain CDR3 in 5 normal blood donors and the dominant CDR3 in the PBMCs in 5 SLE patients. Sequence analysis of the CDR3 region in monoclonal or oligoclonal T cells was performed.

RESULTSThe spectratypes of TCR BV gene CDR3 region showed Gaussian distribution in the 5 normal blood donors. The 5 SLE patients, however, displayed anomalous proliferation and oligoclonal expansion of the T cells was observed in different TCR BV families with different CDR3 sequences.

CONCLUSIONNoticeable drift of TCRbeta chain CDR3 can be seen in active SLE, indicating possible association of selective expression of TCR with immune pathogenesis in SLE. Determination of specific TCR CDR3 sequence provides a new means for studying the pathogenesis and personalized treatment of SLE.

Amino Acid Sequence ; Base Sequence ; Cloning, Molecular ; Complementarity Determining Regions ; genetics ; Female ; Genes, T-Cell Receptor beta ; genetics ; Humans ; Leukocytes, Mononuclear ; cytology ; metabolism ; Lupus Erythematosus, Systemic ; genetics ; immunology ; Molecular Sequence Data ; Sequence Analysis, DNA

INTRODUCTIONBased on the estimate results of the capacity and preparedness of Beijing hospitals to respond to pandemic influenza, using flu surge model to evaluate its applicable hypothesis and to provide government with sentient strategy in planning pandemic influenza. Through collection of medical resources information, we calculated the possible impaction on hospitals by Flu Surge model and explored the applicable hypothesis in model operation through a questionnaire, direct observation and group discussion in 3 hospitals in Beijing. Based on flu surge model estimation during a 6-week epidemic from a pandemic virus with 35% attack rate, Beijing would have had an estimation of 5 383 000 influenza illnesses, 2 691 500 influenza outpatients, 76 450 influenza hospitalizations and 14 508 excess deaths. For a 6-week period with 35% attack rate, there would be a peak demand for 8% of beds, 210% of ICU beds, and 128% of ventilators estimated. Outpatients in different level hospital were quite disproportionated with 1742/ hospital/day, 650/hospital/day, and 139/hospital/day respectively. The sampled health workers had a mastery of 63.4% of the total knowledge and skills of diagnosing and treating of influenza, 73.5% of them washed their hands and 63.5% used PPE correctly. The total beds capacity, medical beds capacity and respiratory medical beds capacity would increase 8%, 35% and 128% respectively.

CONCLUSIONThe estimation results could be referenced when planning the pandemic strategy, but the results should be treated objectively when considering the hypothesis and practical situation in this model being used.

Adolescent ; Adult ; Aged ; Child ; Child, Preschool ; Disease Outbreaks ; statistics & numerical data ; Female ; Hospital Bed Capacity ; Hospital Planning ; Hospitalization ; statistics & numerical data ; Humans ; Infant ; Infant, Newborn ; Influenza, Human ; epidemiology ; Male ; Middle Aged ; Models, Statistical ; Surge Capacity ; Young Adult

BACKGROUNDTo analyze the genetic polymorphism of HLA-Cw locus in Chinese Yi ethnic group by DNA typing for further study on its association with HIV infection and progression to AIDS.

METHODSA rapid genotyping method for HLA-Cw by PCR-SSP was set up. It combined twenty-six specific primers and one pair of internal control primer to form twenty-four one-step reactions for each sample. Totally 102 unrelated healthy Chinese Yi ethnic individuals were typed.

RESULTSTwelve HLA-Cw alleles were detected in Chinese Yi ethnic group with HLA-Cw*01, Cw*07 and Cw*08 as the most common genes, which accounted for a frequency of 0.333 3, 0.250 0 and 0.176 5 respectively; four kinds of non-serologically defined HLA-Cw genes i.e. Cw*12, Cw*1301, Cw*14 and Cw*15 were found in this population. Hardy-Weinburg test showed that the genotype distribution observed was correspondent with the expected (chi2=65.983 1, df=66, P>0.05).

CONCLUSIONSThis study provides the data of HLA-Cw gene frequency in Chinese Yi ethnic group, which may contribute to research on anthropology, disease association and vaccine application. The result also confirmed that PCR-SSP was a reliable and fast method for HLA-Cw genotyping.

Alleles ; Asian Continental Ancestry Group ; genetics ; China ; ethnology ; Gene Frequency ; Genotype ; HLA-C Antigens ; genetics ; Humans ; Polymerase Chain Reaction ; methods ; Polymorphism, Genetic ; Random Allocation

OBJECTIVETo investigate the human immunodeficiency virus (HIV) and syphilis incidence rates as well as the retention rate in a cohort with 6-month follow-up study among female sex workers (FSWs).

METHODSFrom December, 2004, to January, 2005, a community-based baseline survey was conducted to recruit 343 FSWs for a prospective cohort study in Xichang county of Sichuan province, China. Follow-up visits were conducted at 6 months to analyze risk factors associated with cohort retention for subjects' baseline sociodemographic and sexual behavioral characteristics. Blood specimens were also collected to test antibodies against HIV and syphilis.

RESULTSDuring the 6-month follow-up period, HIV and syphilis incidence appeared to be 1.00 per 100 person-years and 6.23 per 100 person-years, respectively. The rate of retention in the cohort was 53.6% (184/343). Results from multivariate logistic regression model showed that factors were significantly associated with cohort retention including people with minority ethnic background (OR = 0.36; 95% CI: 0.18-0.74), people having participated in AIDS prevention program (OR = 1.83; 95% CI: 1.17-2.86) or being clients in the last 6 months > or = 50 (OR = 1.75; 95% CI: 1.11-2.77) and having changed living/working place (OR = 0.56; 95% CI: 0.33-0.94).

CONCLUSIONThe results of this study showed that the syphilis incidence and unprotected sex behavior were high among local FSWs. People belonged to Han nationality, having participated in AIDS prevention program and having a steady living/working place were associated with cohort retention at 6-month follow-up study among FSWs, respectively.

China ; epidemiology ; Female ; Follow-Up Studies ; HIV Infections ; epidemiology ; Humans ; Incidence ; Risk Factors ; Sex Work ; Sexual Behavior ; Substance-Related Disorders ; Syphilis ; epidemiology

OBJECTIVETo explore genetic polymorphisms CCR5 of HIV coreceptor and CCR2 in Chinese Yi Ethnic group in Sichuan.

METHODSGenomic DNA samples were obtained from 119 healthy individuals and 88 HIV-1 infected individuals of Chinese Yi Ethnic group in Sichuan. Polymerase chain reaction (PCR), cloning and gene sequencing techniques were employed to identify the genotype of CCR5Delta32; PCR-restriction fragment length polymorphism (RFLP) and gene sequencing were employed to identify the CCR2-64I alleles.

RESULTSAt CCR5 locus, 2 heterozygotes (CCR5-wt/Delta32) and none homozygote (CCR5-Delta32/Delta32) were observed in 119 healthy individuals, allelic frequency of CCR5-Delta32 was 0.84%; No mutant was found in 88 HIV-1 infected individuals. At CCR2 locus, 26 heterozygotes (CCR2-64V/64I) and two homozygotes (CCR2-64I/64I) were observed in healthy individuals but the allelic frequency CCR2-64I was 12.61%. Among infected individuals, 12 heterozygotes (CCR2-64V/64I) and 7 homozygotes (CCR2-64I/64I) were observed and the allelic frequency CCR2-64I was 13.27%. Statistical analysis revealed that the differences of both loci between healthy and infected individuals were insignificant. Both loci were consistent with the Hardy-Weinberg equilibrium in the two different groups.

CONCLUSIONThe polymorphism of CCR5Delta32 and CCR2-64I alleles from Chinese Yi Ethnic group was detected which was of significance for the evaluation of genetic resistance to HIV-1 infection in Chinese population.

Adult ; Alleles ; Base Sequence ; China ; ethnology ; Female ; HIV Infections ; virology ; HIV-1 ; genetics ; Humans ; Male ; Middle Aged ; Molecular Sequence Data ; Polymorphism, Genetic ; Receptors, CCR5 ; genetics ; Receptors, HIV ; genetics

OBJECTIVETo investigate the clinical therapeutic effect of methylprednisolone combined with cyclophosphamide and Etanercept method on acute paraquat poisoning.

METHODS136 patients with acute paraquat poisoning were divided into the normal therapy group and the intensive therapy group randomly. Methylprednisolone, cyclophosphamide and Etanercept were used in the intensive therapy group. Methylprednisolone 500 mg was given intravenously per day for continuous three days followed by 200 mg intravenous per day. Then methylprednisolone was decreased gradually 14 d or 21 d later according to the patient's condition. Cyclophosphamide 600 mg was given intravenously twice weekly for 2 weeks and Etanercept 25 mg was given hypodermic injection twice weekly for 3 weeks. Curative effect evaluation was done at 7, 14, 21 d and 12 weeks after therapy.

RESULTSThe survival rate of the intensive therapy group was obviously higher than that of the normal therapy group (P<0.01) on 7, 4, 21 d and 12 weeks. The cure rate of the intensive group were 94.6% (intake dose<50 ml 20% paraquat solution), 75.0% (intake dose 50 approximately 100 ml 20% paraquat solution), 12.5% (intake dose>100 ml 20% paraquat solution) respectively, while the cure rate of the normal group were 16.7% (intake dose<50 ml 20% paraquat solution), 8.3% (intake dose 50 approximately 100 ml 20% paraquat solution), 0% (intake dose>100 ml 20% paraquat solution) respectively. The total cure rate of the intensive therapy group (78.3%) 12 weeks later was higher than that of the normal group (11.9%).

CONCLUSIONMethylprednisolone combined with cyclophosphamide and Etanercept intensive therapy has the curative effect on acute paraquat poisoning.

Acute Disease ; Adolescent ; Adult ; Cyclophosphamide ; administration & dosage ; therapeutic use ; Drug Therapy, Combination ; Etanercept ; Female ; Humans ; Immunoglobulin G ; administration & dosage ; therapeutic use ; Male ; Methylprednisolone ; administration & dosage ; therapeutic use ; Middle Aged ; Paraquat ; poisoning ; Poisoning ; drug therapy ; Receptors, Tumor Necrosis Factor ; administration & dosage ; therapeutic use ; Treatment Outcome ; Young Adult

OBJECTIVE: To investigate the efficacy of Bushen Huoxue Fang (BSHXF, a traditional Chinese medicine formula) for improving recurrent spontaneous abortion (RSA) in mice and the role of tyrosine kinase (JAK2) and transcriptional activator (STAT3) signaling pathway in its therapeutic mechanism. METHODS: Female CBA/J mice were caged with male DBA/2 mice to establish RSA mouse models, which were randomly divided into model group, dydrogesterone group and BSHXF group, with the female mice caged with male BALB/c mice as the control group (n=6). From the first day of pregnancy, the mice were subjected to daily intragastric administration of BSHXF, dydrogesterone, or distilled water (in control and model groups) for 12 days. After the treatments, serum levels of antithrombin III (AT-III), activated protein C (APC), tissue plasminogen activator (t-PA), progesterone, human chorionic gonadotropin (HCG), and estradiol (E2) were detected in each group using ELISA. HE staining was used to observe the morphological changes of the endometrium of the mice. Western blotting was performed to determine the expressions of p-JAK2, p-Stat3 and Bcl-2 in the placenta of the mice. RESULTS: Compared with the control mice, the mouse models of RSA showed a significantly increased embryo loss rate with decreased serum levels of AT-III, T-PA, progesterone, APC and HCG, increased placental expressions of p-JAK2, p-STAT3 and Bax, and decreased expression of Bcl-2 (P < 0.05). Treatments with BSHXF and dydrogesterone both increased serum levels of AT-III, t-PA and HCG in the mouse models; Serum APC level was significantly reduced in BSHXF group and serum progesterone level was significantly increased in dydrogesterone group (P < 0.05). CONCLUSION BSHXF can improve the prethrombotic state and inhibit cell apoptosis by downregulating the JAK2/STAT3 pathway to increase the pregnancy rate in mouse models of RSA.
Animals ; Mice ; Abortion, Habitual/prevention & control* ; Signal Transduction ; Down-Regulation ; Disease Models, Animal