Objective To construct recombinant plasmid with p24 and gp41 gene, express fusion protein in E.coli. Methods To design primer with restriction endonuclease position,and amplify p24 and gp41 by RT-PCR, link both into pMD18-T vector.To choose correct clone with target gene.Then p24 fragment will be cleaved and linked into pMD18-T vector within gp41 gene. Both post-linked gene will be cleaved and linked into pET21a vector. The vector will be transformed into E.coli. And protein is highly effective expressed in E coli. Western blotting proved that the expressed product could react with 6?his antibody. Result Fusion protein p24-gp41 is highly effective expressed in E.coli. Conclusion Fusion protein p24-gp41 is highly effective expressed in E.coli in pET21a vector.

Objective To comprehend the current difference of sequence variation of p24 coding region in gag gene and subtype distribution of HIV 1 in Henan province and Shanghai in China. Methods Plasma specimens were collected from 37 HIV 1 patients, of which 25 were from Henan province and 12 were from Shanghai. Most patients in Henan were infected by illegally donating blood while in Shanghai the major transmission routes were blood or blood product transfusion and sexuality. RNAs from plasma specimens were extracted and target gene were amplified by the method of RT PCR and Nested PCR. The sequences of p24 region, 693 nucleotides were determined, then phylogenetic analyses were performed. Results Subtyping showed that 83.8% (31/37) were B subtype. Of 25 Henan specimens, 23(92%)were B subtype and 2 were A subtype. Of 12 Shanghai specimens, 8(66.7%)were B subtype, 1 was A subtype, 2 were CRF01 -AE and 1 was CRF02 -AG. Comparing with the Consensus sequence (Consensus -B, from HIV database), nucleotide variation in B subtype of Henan was 1.6%～4.2%, with an average of 3.2%, while that of Shanghai was 2.0%～3.8%, with an average of 3.4%. No G to A hypermutation was observed in all variations. The median intrasubtype distance for B subtype in Henan and Shanghai was 2.9% and 3.5%, and the intersubtype distance between B subtype and other subtypes was 11.1%～12.5%. The two Consensus sequences of Henan and Shanghai B subtype obtained by CLUSTAL X were aligned to Consensus -B, both the substitution for predicted amino acid were 2.2%(5/231). Of all the variations,they shared the same three mutations: A14P、I91V and E180D. However, the disparity between the two Consensus sequences was not significant (0.9%). Phylogenetic analyses implied that many specimens were B subtype, and all the B subtype specimens from Henan and most of Shanghai B subtype specimens were close to the isolates in Thailand. Conclusion B subtype was the dominant HIV 1 isolate in Henan and Shanghai of China.The HIV 1 B subtype in Henan and Shanghai had coincident homology and shared some identical variations of amino acid.

Objective To quantitatively investigate the expression levels of the cytosine deaminases,hA3B,hA3F and hA3G in peripheral blood mononuclear cell(PBMC)of human immunodeficiency virus(HIV)infected patients and to analyze the correlation between cytosine deaminases expression and CD4~+ T lymphocyte counts. Methods Peripheral blood samples were collected from 21 HIV-infected subjects who didn't take antiretroviral therapy(ART-),21 HIV-infected subjects receiving ART(ART+),and 10 HIV-uninfected subjects. PBMC were isolated by Ficoll density gradient centrifugation, followed by RNA extraction and cDNA synthesis.hA3B,hA3F and hA3G mRNA levels were determined by real-time fluorescent quantitative polymerase chain reaction(PCR).CD4~+ T lymphocyte counts were determined using flow cytometry. Data were analyzed by t test, t' test or Wilcoxon rank sum test. Results In HIV-infected subjects without or with ART,HIV-uninfected subjects, the levels of hA3B mRNA were 208.4,365.2 and 563.6,hA3F mRNA were 245.5,316.6 and 442.9,hA3G mRNA were 404.6,360.8 and 638.6,respectively.hA3G mRNA level in HIV-infected subjects was lower than that in HIV-uninfected controls(P=0.0131),but there was no statistical difference between ART+ and ART-groups(P=0.7342).There were no correlations between hA3B,hA3F and hA3G mRNA levels and CD4+ T lymphocyte counts in either ART-or ART+HIV-infected subjects(ART-:r=-0.0104,r=-0.0545,r=0.1623,all P＞0.05;ART+:r=0.3220,r=0.2193,r=0.1455,all P＞0.05).In addition,hA3B,hA3F and hA3G mRNA levels were positively correlated with one another in ART-HIV-infected subjects and HIV-uninfected controls(P＜0.05),but not in ART-HIV-infected subjects(P＞0.05).Conclusions hA3B,hA3F and hA3G expression levels do not directly correlate with CD4~+ T lymphocyte counts in HIV-1-infected patients,hA3B,hA3F and hA3G expression levels in PBMCs tend to he decreased after HIV-1 infection, and ART may increase hA3B and hA3F mRNA expression.

ObjectiveTo study the clinical value and efficacy of intracranial hemorrhage smashpuncture needle with YL-1 type disposable under CT monitoring in treatment of hypertensive intracerebral hemorrhage.MethodsThe data of 80 cases with hypertensive intracerebral hemorrhage patients who accepted the transcranial puncture under CT monitoring application of YL-1 type disposable intracranial hemorrhage smash puncture needle and drainage of hemorrhage,punctuated with urokinase washout and drainaged residual blood clot.Evaluation criteria:hemorrhage volume reduction,average operation time,preoperative and postoperative Glasgow coma scale(GCS) score,the clinical effect of long-term follow-up.ResultsOperation time was 15-45 (25.0 ± 2.8) minutes;hemorrhage volume was reduced by an average of 30％-80％ (56.8 ± 3.2)％,the average increase of GCS was(2.3 ±0.3) scores,10 cases death in 80 patients,70 survivors' activities of daily living (ADL) assessments:grade ADL 1 in 17 cases ( 24.3 ％ ),grade ADL2 in 36 cases( 51.4％ ),grade ADL3 in 13 cases ( 18.6％ ),grade ADL4 in 3 cases(4.3％ ),grade ADL5 in 1 case ( 1.4％ ).ConclusionsYL-1 type disposable intracranial hemorrhage smash puncture needle under CT monitoring in the treatment of hypertensive intracerebral hemorrhage is a simple,fast and accurate positioning,without craniotomy and blood transfusion,safe and effective operation,but should pay attention to operation indications.

Objective To analyze the causes of death for 89 HIV/AIDS patients,and the association of CD4+ T lymphocyte counts with the mortality.Methods Data were collected from 89 deceasedpatients who were admitted to Shanghai Public Health Clinical Center from Jan 1996 to Sep 2007,and wereretrospectively analyzed with t-test.Results Heterosexual,blood transfusion and blood products were themajor transmission routs for 89 deceased HIV/AIDS patients.The primary causes of death varied with different levels of CD4+ T lymphocyte counts.With higher CD4+T lymphocyte counts,patients were more likelyto die from non-AIDS-related opportunistic infections such as upper digestive tract hemorrhage.Serious pulmonary infection induced by various etiologies was the primary cause of death.Co-infections of multiple etiologics were often seen in these death cases.Conclusion Opportunistic infection is the primary cause of death for HIV/AIDS patients.Appropriate therapies should be based on CD4+ T lymphocyte counts and patients' specific conditions to reduce the mortality.

Objective To elavluate the efficacy of pelvic floor muscle training (PFMT) on symptoms and quality of life in the treatment of female overactive bladder (OAB).Methods Ninety-one female patients with OAB completed the prospective study and were divided into 2 groups.The study group had PFMT combined with comprehensive health education and tolterodine (n =46),and the control group had comprehensive health education combined with tolterodine (n =45).OAB symptom score (OABSS),King's health questionnaire (KHQ) and modified Oxford scale (MOS) were evaluated at baseline,2 weeks,1 month and 3 months.The patients were followed up for 6 months.Results The MOS and OABSS score in study group before the treatment were 2.87±0.65 and 7.61 ±2.28,respectively,and in control group they were 2.80±0.55 and 7.44±2.41,respectively.There was no significant difference between the 2 grotps (P＞ 0.05).There was no significant difference in KHQ domains before the treatment between the 2 groups (P＞ 0.05).The MOS scores were 3.15±0.63 and 3.57±0.58 after treatment of 1 and 3 months in study group,which were significantly improved compared with the values of 2.89±0.57 and 3.09±0.67 in control group (P＜0.05).The OABSS score was 1.13± 1.93 in study group after treatment of 6 months,which was significantly improved compared with the values of 2.47±2.18 in control group (P＜0.05).The Incontinence impact,Role Limitations,Physical Limitations,Social Limitations and Symptom Severity in study group improved significantly compared with the values in control group after treatment of 3 months (P＜0.05).Domains of KHQ scores were totally significantly improved after treatment of 6 months in study group than in control group(P＜0.05).Conclusion Long term PFMT may play an important role in the treatment of OAB,and can also improve the quality of life of the patients.

Objective: To explore window settings technology in the digital radiography of pneumoconiosis, and to improve the quality of the digital chest radiographs. Methods: 25 female workers in a asbestos product processing enterprise were examined with high-kV and DR chest radiographies on same day. Consistency of pneumoconiosis diagnosis results: were assessed. Results In the 75 groups of radiograph quality results, there are 62 groups that need to be processed window in order to make the correct diagnosis. The crude agreement among two chest radiographs was 28% for the profusion of small opacities and 76% for the classification of pneumoconiosis, the weighted kappa value (κ) was 0.33 (95%CI: 0.12-0.54) and 0.67 (95%CI: 0.42-0.92) . Conclusions window settings technology can display different parts of chest and make DR chest radiograph to meeting the requirements of chest radiograph quality for pneumoconiosis.