[Epidemiological distribution of genotypes and sub-genotypes of hepatitis B virus in 15 ethnic groups in China].

Objective: To understand the distribution of genotypes and sub-genotypes of HBV in different ethnic groups in China. Methods: The HBsAg positive samples were selected by stratified multi-stage cluster sampling from the sample base of national HBV sero-epidemiological survey in 2020 for the amplification of S gene of HBV by nested PCR. A phylogeny tree was constructed to determine the genotypes and sub-genotypes of HBV. The distribution of genotypes and sub-genotypes of HBV were analyzed comprehensively by using laboratory data and demographic data. Results: A total of 1 539 positive samples from 15 ethnic groups were successfully amplified and analyzed, and 5 genotypes (B, C, D, I and C/D) were detected. The proportion of genotype B was higher in ethnic group of Han (74.52%, 623/836), Zhuang (49.28%, 34/69), Yi (53.19%, 25/47), Miao (94.12%, 32/34), Buyi (81.48%, 22/27). The proportions of genotype C were higher in ethnic groups of Yao (70.91%, 39/55). Genotype D was the predominant genotype in Uygur (83.78%, 31/37). Genotype C/D were detected in Tibetan (92.35%,326/353). In this study, 11 cases of genotype I were detected, 8 of which were distributed in Zhuang nationality. Except for Tibetan, sub-genotype B2 accounted for more than 80.00% in genotype B in all ethnic groups. The proportions of sub-genotype C2 were higher in 8 ethnic groups, i.e. Han, Tibetan, Yi, Uygur, Mongolian, Manchu, Hui and Miao. The proportions of sub-genotype C5 were higher in ethnic groups of Zhuang (55.56%, 15/27) and Yao (84.62%, 33/39). For genotype D, sub-genotype D3 was detected in Yi ethnic group and sub-genotype D1 was detected in both Uygur and Kazak. The proportions of sub-genotype C/D1 and C/D2 in Tibetan were 43.06% (152/353) and 49.29% (174/353). For all the 11 cases of genotype I infection, only sub-genotype I1 was detected. Conclusions: Five genotypes and 15 sub-genotypes of HBV were found in 15 ethnic groups. There were significant differences in the distribution of genotypes and sub-genotypes of HBV among different ethnic groups.

[Self-awareness rate and its influencing factors of their infection status among hepatitis B surface antigen-positive persons aged 15-69 years in China].

Objectives: To understand the awareness rate and its influencing factors of their HBV infection status among HBsAg-positive persons aged 15-69 years in China. Methods: A cross-sectional design was used to conduct a questionnaire survey on the awareness of their infection status among HBsAg-positive persons aged 15-69 years who were identified in the 2020 national hepatitis B seroepidemiology survey. The awareness rate of the whole respondent and respondents with different characteristics were described, and the differences were compared with the χ2 test. The logistic regression model was used to analyze the factors influencing the awareness rate. Results: The overall awareness rate among the respondents was 43.10% (1 828/4 241). The awareness rate was lower in males than in females (41.30% vs. 44.65%). The awareness rate was lower in the 60-69-years-old age group than in other age groups (30.38% vs. 36.77%-57.58%). The awareness rate was lower in rural areas than in urban areas (39.43% vs. 47.32%). The awareness rate was lower in regions with a per capita gross domestic product (GDP) below RMB 54 000 than in regions with a per capita GDP of RMB 54 000 and above (36.81% vs. 41.61%-50.30%). The awareness rate was lower in respondents without other liver diseases than with other liver diseases (41.52% vs. 60.68%). The awareness rate was lower in respondents without a family history of hepatitis B-related disease or unknown family history than with a family history (43.58% vs. 68.26%; 24.71% vs. 68.26%). Multivariate logistic regression analysis showed that male [odds ratio (OR)=0.841, 95% confidence interval (CI): 0.734-0.964], high school and below [primary school and below, junior middle school, high school/technical secondary school, OR (95%CI): 0.247 (0.190-0.321), 0.451 (0.352-0.577), 0.634 (0.486-0.827)], rural areas (OR=0.822, 95%CI: 0.715-0.945) and regions with a per capita GDP below RMB 80 000 [54 000-80 000, OR (95%CI): 0.810 (0.688-0.954), below RMB 54 000, OR (95%CI): 0.793 (0.669-0.941)] were the negative factors influencing the awareness rate. While 30-39-years-old (OR=2.089, 95%CI: 1.626-2.683) and 40-49-years-old (OR=1.590, 95%CI: 1.250-2.023) age groups, with other liver diseases (OR=2.244, 95%CI: 1.754-2.871) and family history related to hepatitis B (OR=2.688, 95%CI: 2.242-3.223) were the positive factors influencing the awareness rate. Conclusion: The overall awareness rate of their infection status among HBsAg-positive persons aged 15-69 years is 43.10% in China. Health promotion and coverage expansion on HBV screening should be further strengthened to achieve the proposed World Health Organization's target of 90% HBV infection diagnosis rate by 2030.

[Sero-epidemiological characteristics of hepatitis E in population aged 1-29 years in China, 2014].

Objective: To analyze the epidemiological characteristics of hepatitis E Virus antibody (anti-HEV) in people aged 1-29 years in China in 2014. Methods: Based on database of the national serologic survey of hepatitis B in people aged 1-29 years in China, in 2014, the sample size was estimated. The serum samples of the people surveyed were randomly selected to detect anti-HEV IgG by using enzyme- linked immunosorbent assay (ELISA). Statistical software SAS 9.1.3 was used to calculate the positive rate of anti-HEV and 95% confidence interval (CI) in different age, gender groups, urban and rural areas and geographic areas by using the Taylor series linear method with sampling weight. The difference was determined by comparing 95%CI. Results: A total of 14 106 serum samples were detected from people aged 1-29 years, including 6 996 males (49.60%), 7 013 urban residents (49.72%). The positive rate of anti-HEV was 8.12%(95%CI: 7.19-9.15) in people aged 1-29 years. There was no statistical difference between the men and women, between urban area and rural area. The positive rates of anti-HEV in western area(11.36%, 95%CI: 9.45-13.62) was higher than those in eastern and central areas. The positive rates of anti-HEV were 2.46%, 2.24%, 4.50%, 7.58%, 11.89% and 17.27% in people aged 1-, 5-, 10-, 15-, 20- and 25-29 years, respectively. As the age increased, the positive rate of anti-HEV gradually increased. In different ethnic groups, the positive rate of anti-HEV was higher in Tibetan (18.32%, 95%CI: 12.02-26.90), Zhuang (9.54%, 95%CI: 4.33-19.73) ethnic groups. Conclusion: The positive rate of anti-HEV declined slightly in China in 2014. It is still necessary to pay close attention to the HEV infection, morbidity of hepatitis E and risk factors in people aged 1-29 years.

[Differentiation of prostatitis and clinically significant prostate cancer in peripheral zone using the prostate imaging-reporting and data system].

Objective: To determine whether clinically significant prostate cancer (PCa) and prostatitis in the peripheral zone can be distinguished using prostate imaging reporting and data system version 1 (PI-RADS V1) and version 2(PI-RADS V2). Methods: Between September 2010 and August 2016, mpMRI data of 77 patients with PCa and 29 prostatitis obtained at 3.0 T were collected in Zhangjiagang Hospital Affiliated to Soochow University. Every lesion was scored according to PI-RADS (V1 and V2), as well as a sum score and a PI-RADS V2 score. The non-parametric Kruskal-Wallis test was used to assess differences between PCa and prostatitis regarding PS3, PS4 and PI-RADS V2 score. The diagnostic performance of PI-RADS V1 and V2 for detection of prostatitis in peripheral zone was compared by analyzing ROC curve. Results: The PI-RADS V1 score for PS3, PS4 and the PI-RADS V2-score were all significantly higher for PCa (PS3:12.1±2.1; PS4:16.2±2.9; V2:4.6±0.8) than for prostatitis (PS3:8.0±0.7; PS4:10.6±1.0; V2:3.0±0.5) (all P<0.01). Of these parameters, PS4 achieved the highest predictive value for the presence of prostatitis with an AUC of 0.937, sensitivity and specificity were 87.0%, 97.0% with a threshold of 12.5. Conclusion: Prostatitis can be differentiated from clinically significant PCa in peripheral zone on mpMRI using PI-RADS system, PS4 achieved better results compared to PS3 and V2.

[Comparative analyze on hepatitis B seroepidemiological surveys among population aged 1-29 years in different epidemic regions of China in 1992 and 2014].

Objective: To evaluate the effect of hepatitis B prevention and control by comparative analysis on the results of HBsAg, anti-HBs and anti-HBc prevalence from national hepatitis B seroepidemiological surveys in 1992 and 2014 in different epidemic regions of China. Methods: Data was from the national seroepidemiological surveys of hepatitis B conducted in 1992 and 2014. The survey in 1992 was conducted in 145 disease surveillance points of 30 provinces (excluding Hong Kong, Macao Special Administrative Region and Taiwan province) in China. The survey in 2016 was conducted in 160 disease surveillance points of 31 provinces (excluding Hong Kong, Macao Special Administrative Region and Taiwan province) in China. In the two surveys, face-to-face interviews with the subject by door to door or on the investigation site were conducted by trained staff using standard questionnaires to obtain basic information including birth date, gender, ethnicity, resident place and so on. And then 5 ml venous blood was collected to test the sero-markers of HBsAg, anti-HBs and anti-HBc. We analyzed unweighted point prevalence and 95% CI of HBsAg, anti-HBs and anti-HBc in 1992 which had no design weighting, and analyzed weighted point prevalence and 95%CI of HBsAg, anti-HBs and anti-HBc in 2014 which had design weighting. Results: 34 291 and 31 713 people aged 1-29 years were involved in 1992 and 2014 national serosurveys of China, respectively. For the people aged 1-29 years, HBsAg prevalence was 2.64% (95%CI: 2.28%-3.06%) in 2014 and decreased by 73.92% as compared with the rate 10.13% (95% CI: 9.81%-10.45%) in 1992. Anti-HBc prevalence was 13.01% (95%CI: 12.09%-14.00%) in 2014 and decreased by 71.61% as compared with the rate 45.84% (95% CI: 45.31%-46.37%) in 1992. Anti-HBs prevalence was 57.79% (95%CI: 56.33%-59.25%) in 2014 and ascended by 127.41% as compared with the rate 25.41% (95% CI: 24.95%-25.87%) in 1992. In high, medium and low epidemic region, for the people who born during 1992-2001 when hepatitis B vaccine was introduced in routine immunization management, HBsAg prevalence was 4.74% (95%CI: 3.79%-5.69%), 1.59% (95%CI: 1.09%-2.10%) and 2.53% (95%CI: 1.66%-3.39%), respectively, and anti-HBs prevalence was 64.25% (95% CI: 62.11%-66.39%), 56.34% (95% CI: 54.50%-58.57%), 54.49% (95%CI: 51.75%-57.23%), respectively, and anti-HBc prevalence was 15.16% (95%CI: 13.56%-16.76%), 11.07% (95%CI: 9.80%-12.33%), 7.61% (95%CI: 6.15%-9.07%), respectively. In high, medium and low epidemic region, for the people who born during 2002-2013 the duration which hepatitis B vaccine was integrated in expanded immunization program born during when HBsAg prevalence was 0.88% (95%CI: 0.66%-1.11%), 0.37% (95%CI: 0.24%-0.49%)and 0.71% (95%CI: 0.48%-0.94%), respectively, and anti-HBs prevalence was 60.74% (95%CI: 59.57%-61.90%), 59.46% (95%CI: 58.44%-60.49%), 52.56% (95% CI: 51.20%-53.92%), respectively, and anti-HBc prevalence was 3.30% (95% CI: 2.87%-3.72%), 1.91% (95%CI: 1.63%-2.20%), 2.25% (95%CI: 1.85%-2.66%), respectively. Conclusion: China had made great achievement in hepatitis B prevention and control. HBsAg prevalence among people aged 1-29 years old in 2014 decreased dramatically as compared with that in 1992. Since hepatitis B vaccine was integrated into expanded immunization program, China reduced HBsAg prevalence to less than 1% among people aged 1-12 years in 2014 in different epidemic region.

[Epidemiological characteristics of children aged 1-4 years without timely birth dose of hepatitis B vaccine vaccination in China, 2014].

Objective: To evaluate the epidemiological characteristics of the children aged 1-4 years without timely birth dose of hepatitis B vaccine (HepB(1)) vaccination. Methods: Based on the data from 160 disease surveillance points in 31 provinces of China, two-stage cluster random sampling was used to select the target population aged 1-4 years. A standard questionnaire was used to collect the information about the birth date, gender, ethnic group, place of birth, HepB immunization history of the children selected. A blood sample (3 ml) was taken from each subject for HBsAg testing. SAS software (Version 9.4) was used in our study. We analyzed the age, gender, ethnic group, area specific distributions of the children aged 1-4 years without timely HepB(1) vaccination and the influencing factors, and the relationship between the HepB(1) vaccination time and HBsAg prevalence rate. Results: A total of 12 587 children aged 1-4 years were analyzed and the non-timely HepB(1) vaccination rate was 10.12%. The place of birth, ethnic group, urban/rural area, eastern/central/western area, age were the main influencing factor of the non-timely HepB(1) vaccination. The non-timely HepB(1) vaccination rate was higher in 3-4 years old children (11.13%) than in 1-2 years old children (8.97%), in rural area (12.05%) than in urban area (8.19%), in western area (13.41%) than in central area (9.27%) and eastern area (7.72%), in minority ethnic group (18.06%) than in Han ethnic group (8.77%) and in children born outside hospital (57.66%) than in children born in hospital (9.27%). The HBsAg prevalence rate among 1-4 years children was 0.31%. The HBsAg prevalence rate of the children with timely HepB(1) vaccination (0.25%) was lower than that of the children without timely HepB(1) vaccination (0.89%). Conclusions: In China, the HBsAg prevalence rate among 1-4 years children with HepB vaccination decreased to <0.5% and the timely HepB(1) vaccination rate reached to 90%. We should strengthen the timely HepB(1) vaccination for the children in minority ethnic groups, in western area, in rural area as well as those born outside hospitals.

Long-term effectiveness of plasma-derived hepatitis B vaccine 22-28 years after immunization in a hepatitis B virus endemic rural area: is an adult booster dose needed?

Longan County is considered a highly endemic area for hepatitis B virus (HBV). The plasma-derived vaccine has been used in newborns in this area since 1987. A cross-sectional survey was conducted to evaluate the long-term effectiveness of this vaccine. In total, 1634 participants born during 1987-1993 and who had received a series of plasma-derived HB vaccinations at ages 0, 1, and 6 months were enrolled. Serological HBV markers were detected and compared with previous survey data. Overall the prevalence of hepatitis B surface antigen (HBsAg) in all participants was 3·79%; 3·47% of subjects who had received the first dose within 24 h were HBsAg positive, and 8·41% of subjects who had received a delayed first dose were also HBsAg positive. There were 1527 subjects identified who had received the first dose within 24 h and whose HBsAg and anti-HBc prevalence increased yearly after immunization, while the anti-HBs-positive rate and vaccine effectiveness declined. The geometric mean concentration of antibody in the anti-HB-positive participants was 55·13 mIU/ml and this declined after immunization. Fewer than 2·0% of participants had anti-HB levels ⩾1000 mIU/ml. The data show that the protective efficacy of the plasma-derived vaccinations declined and administration of HB vaccine within 24 h of birth was very important. To reduce the risk of HBV infection in this highly endemic area, a booster dose might be necessary if anti-HBs levels fall below 10 mIU/ml after age 18 years. Furthermore, studies on the immune memory induced by plasma-derived HB vaccine are needed.

[Effect of traditional Chinese medicine treatment as maintenance therapy on regulating the serum concentration of sCTLA-4 in patients with advanced non-small-cell lung cancer and its relationship with prognosis].

Objective: To evaluate the effect of traditional Chinese medicine (TCM) treatment as maintenance therapy on regulating the serum concentration of soluble cytotoxic T lymphocyte associated antigen-4 (sCTLA-4) in patients with advanced non-small-cell lung cancer (NSCLC) and the relationship between sCTLA-4 and time to progression (TTP). Methods: This study was conducted as a prospective, randomized, controlled trial. 64 non-progressive patients who responded to initial therapy were randomized 1∶1 to the TCM arm (treated with cinobufacini injection, herbal decoction and Chinese acupoint application, n=32) or to the chemotherapy arm (n=32). Each cycle was 21 days. Cycles were repeated until disease progression, unacceptable toxicity, or until the patient requested therapy discontinuation.The serum concentration of sCTLA-4 was detected by enzyme linked immunosorbent assay (ELISA) in the 64 patients with advanced NSCLC before and after two cycles of maintenance treatment. Cox regression was used to analyze the relative ratio for the risk of disease progression. Results: After 2 cycles of maintenance TCM treatment, the serum concentration of sCTLA-4 in patients with advanced NSCLC was (12.77±2.37 pg/ml), significantly lower than that before treatment (40.30±10.75)(P=0.013). After 2 cycles of maintenance chemotherapy, the serum concentration of sCTLA-4 was higher than that before treatment, but was not significantly different (44.48±10.12 vs. 46.64±11.21 pg/ml, P=0.612). After 2 cycles of maintenance treatment, TCM treatment can significantly bring down the serum concentration of sCTLA-4 compared to chemotherapy (12.77±2.37 vs. 46.64±11.21 pg/ml, P=0.004). The multivariate analysis indicated that sCTLA-4 levels and treatment regimen were independent prognostic factors for TTP (P<0.05 for both). Conclusions: Regulating the serum concentration of sCTLA-4 may be one of the mechanisms of TCM maintenance treatment of NSCLC. Trial registration Chinese Clinical Trial Register, ChiCTR-TRC-10001017.

[The coverage of hepatitis A vaccine among 2-29 year olds and the reporting incidence of hepatitis A in China, 2014].

OBJECTIVE: To evaluate the hepatitis A vaccine coverage among 2-29 year olds and the reported incidence rates of hepatitis A, in China. METHODS: Based on data from the national sero-survey on hepatitis B in 2014, information on hepatitis A vaccine immunization was collected and the coverage of hepatitis A vaccine was analyzed with SAS software (Version 9.4). Incidence data on hepatitis A was also collected from the National Notifiable Disease Reporting System between 2004 and 2014, and analyzed using the micro-software Excel 2007. RESULTS: Totally, data involving 29 058 people aged 2-29 years were available for analysis and the overall hepatitis A vaccine coverage was 44.6%. The younger the age, the higher the coverage appeared. Among the 2-6 year and the 7-14 year olds, rates of hepatitis A vaccine coverage were 91.2% and 76.0% respectively. From 2004 to 2014, the incidence rates of hepatitis A in the whole population were declining, annually. The incidence rates showed continuously declining as 82.5%, 90.6%, 72.1% among children at the age groups of 2-6 years, 7-14 years and in the whole population, from 2007 to 2013. CONCLUSION: After the inclusion of hepatitis A vaccine into the Expanded Programe on Immunization (EPI), the coverage of hepatitis A vaccine among the 2-6 year olds increased to over 90%, with no obvious difference between the urban and rural areas. Incidence of hepatitis A in the 2-6 year olds showed a more rapid decline than that in the whole population.

Epidemiological changes in hepatitis B prevalence in an entire population after 20 years of the universal HBV vaccination programme.

A universal hepatitis B vaccination programme has been conducted in Long An county since 1986. To investigate the epidemiological changes in hepatitis B virus (HBV) infection we conducted a serosurvey there in 2005. A total of 4686 subjects were enrolled and vaccination history and blood samples collected. HBV infective markers were determined by radioimmunoassay. The results were compared with the data of 1985. Our results show that the overall HBsAg prevalence was 7·5%, less than half of the prevalence reported in 1985. HBsAg and anti-HBc antibody prevalence in people born after 1985 decreased markedly. The gender difference in HBsAg prevalence was abolished in subjects aged <20 years. The administration of a first dose of vaccine within 24 h could reduce the HBsAg prevalence by half. In conclusion, the marked epidemiological changes in HBV prevalence found in this serosurvey indicate that the implementation of HBV vaccination was highly successful.

Single-copy gene detection using branched DNA (bDNA) in situ hybridization.

We have developed a branched DNA in situ hybridization (bDNA ISH) method for detection of human papillomavirus (HPV) DNA in whole cells. Using human cervical cancer cell lines with known copies of HPV DNA, we show that the bDNA ISH method is highly sensitive, detecting as few as one or two copies of HPV DNA per cell. By modifying sample pretreatment, viral mRNA or DNA sequences can be detected using the same set of oligonucleotide probes. In experiments performed on mixed populations of cells, the bDNA ISH method is highly specific and can distinguish cells with HPV-16 from cells with HPV-18 DNA. Furthermore, we demonstrate that the bDNA ISH method provides precise localization, yielding positive signals retained within the subcellular compartments in which the target nucleic acid sequences are localized. As an effective and convenient means for nucleic acid detection, the bDNA ISH method is applicable to the detection of cancers and infectious agents. (J Histochem Cytochem 49:603-611, 2001)

[A study on physical characteristics of porous plates for water sublimator].

Objective. To investigate the development method of porous plate, the key part of water sublimator, and its main physical characteristics. Method. Technical parameters of porous plates used in water sublimator were analyzed. On the basis of this, porous plates made of different materials were developed by making use of sintered powder technology. A tester for testing the physical characteristics of porous plate was designed. All around physical characteristic tests for porous plates were conducted. Result. Main factors affecting performance of porous plates were tested. These provided basis for performance test of sublimator, established porous plate criterion and optimized porous plate. Conclusion. Porous plates developed by making use of sintered powder technology could meet requirements of performance of water sublimator.

[Research progress of thermal control system for extravehicular activity space suit].

New research progress of thermal control system for oversea Extravehicular Activity (EVA) space suit is presented. Characteristics of several thermal control systems are analyzed in detail. Some research tendencies and problems are discussed, which are worthwhile to be specially noted. Finally, author's opinion about thermal control system in the future is put forward.

Quantification of cytokine mRNA in peripheral blood mononuclear cells using branched DNA (bDNA) technology.

Changes in the patterns of cytokine expression are thought to be of central importance in human infectious and inflammatory diseases. As such, there is a need for precise, reproducible assays for quantification of cytokine mRNA that are amenable to routine use in a clinical setting. In this report, we describe the design and performance of a branched DNA (bDNA) assay for the direct quantification of multiple cytokine mRNA levels in peripheral blood mononuclear cells (PBMCs). Oligonucleotide target probe sets were designed for several human cytokines, including TNFalpha, IL-2, IL-4, IL-6, IL-10, and IFNgamma. The bDNA assay yielded highly reproducible quantification of cytokine mRNAs, exhibited a broad linear dynamic range of over 3-log10, and showed a sensitivity sufficient to measure at least 3000 molecules. The potential clinical utility of the bDNA assay was explored by measuring cytokine mRNA levels in PBMCs from healthy and immunocompromised individuals. Cytokine expression levels in PBMCs from healthy blood donors were found to remain relatively stable over a one-month period of time. Elevated levels of IFNgamma mRNA were detected in PBMCs from HIV-1 seropositive individuals, but no differences in mean levels of TNFalpha or IL-6 mRNA were detected between seropositive and seronegative individuals. By providing a reproducible method for quantification of low abundance transcripts in clinical specimens, the bDNA assay may be useful for studies addressing the role of cytokine expression in disease.

Critical role of AT1 receptor expression after ischemia/reperfusion in isolated rat hearts: beneficial effect of antisense oligodeoxynucleotides directed at AT1 receptor mRNA.

To examine the relevance of angiotensin II type 1 receptor (AT1R) expression in the determination of myocardial function after ischemia/reperfusion, Sprague-Dawley rats were treated intravenously with antisense oligodeoxynucleotides (AS-ODNs) directed at AT1R mRNA (100 microg/rat, n=9) or scrambled antisense oligodeoxynucleotides (Scr-ODNs, 100 microg/rat, n=6). Both AS-ODNs and Scr-ODNs were given along with 300 microg/rat of liposome DOTAP/DOPE, a positive electron carrier (wt:wt= 1:1). The hearts from AS-ODN- or Scr-ODN-treated rats were excised 24 hours later, perfused in vitro, and subjected to 25 minutes of global ischemia followed by 30 minutes of reperfusion. Parallel groups of rats were given the specific AT1R antagonist losartan (10 mg/kg IV, n=6) or saline (n=7) 4 to 6 hours before excising the hearts. Ischemia/reperfusion resulted in a significant increase in myocardial AT1R expression (autoradiography and binding assay) and myocardial dysfunction, indicated by increases in coronary perfusion pressure and left ventricular end-diastolic pressure and a decrease in developed left ventricular pressure (all P<0.01 versus baseline) in the saline-treated group. AT1R protein and mRNA levels also increased in ischemic/ reperfused myocardial tissues. Administration of AS-ODNs or losartan, but not Scr-ODNs, preserved myocardial function and blocked the increased AT1R binding after ischemia/reperfusion (both P<0.01). Myocardial AT1R mRNA levels were not affected by either AS-ODNs or losartan, and the AT1R protein levels were significantly reduced by AS-ODN, but not losartan, treatment. Plasma angiotensin II levels increased after administration of losartan but not after administration of AS-ODNs. These observations imply a critical role of AT1R upregulation in determining myocardial function immediately after ischemia/reperfusion. AS-ODNs to AT1R mRNA may be more beneficial than losartan, because losartan does not affect the plasma angiotensin II level. The sustained increase in AT1R mRNA, but diminished protein expression, in rat hearts treated with AS-ODNs suggests that AS-ODNs block AT1R at the translational level.

Increased immune activation precedes the inflection point of CD4 T cells and the increased serum virus load in human immunodeficiency virus infection.

The temporal relationship of serum levels of human immunodeficiency virus (HIV) RNA and of immune activation products in 10 HIV-seropositive persons who showed an accelerated decline (inflection point) in CD4 T cell counts and went on to develop AIDS and in 10 matched controls without inflection point were examined. Cases and controls did not differ statistically at the baseline time point for this study. CD4 cell inflection points occurred 18-30 months before AIDS development. Serum levels of soluble tumor necrosis factor receptor II, soluble interleukin-2 receptor, beta2-microglobulin, and neopterin increased significantly > or = 6 months before the CD4 cell inflection point. In contrast, increases in mean HIV RNA levels occurred at the time of the CD4 cell inflection point. These data are consistent with the view that in vivo immune activation precedes the increases in virus load and is followed by an accelerated and rapid loss of CD4 lymphocytes.

Quantification of cytomegalovirus DNA in peripheral blood leukocytes by a branched-DNA signal amplification assay.

Quantification of cytomegalovirus (CMV) DNA in blood may aid in the identification of patients at highest risk for developing CMV disease, the evaluation of new therapeutics, and the prompt recognition of drug-resistant CMV strains. A branched-DNA (bDNA) assay was developed for the reliable quantification of CMV DNA in peripheral blood leukocytes. The bDNA assay allowed for the highly specific and reproducible quantification of CMV DNA in clinical specimens. Furthermore, the bDNA assay was at least as sensitive as culture techniques and displayed a nearly 3 log10 dynamic range in quantification. Changes in CMV DNA levels measured by the bDNA assay in a human immunodeficiency virus-positive patient undergoing therapy were consistent with CMV culture, antigen, and genotype results and correlated with disease progression and resistance markers. The bDNA assay for the quantification of CMV DNA may provide a useful tool that can be used to aid physicians in monitoring disease progression, evaluating therapeutic regimens, and recognizing viral resistance and drug failure.

Diagnosis of cytomegalovirus (CMV) polyradiculopathy and documentation of in vivo anti-CMV activity in cerebrospinal fluid by using branched DNA signal amplification and antigen assays.

Branched chain DNA assay (bDNA), cytomegalovirus (CMV) antigen assay, and cerebrospinal fluid (CSF) viral culture were studied for their utility in the diagnosis of CMV polyradiculopathy and for documenting in vivo antiviral effects. CMV was demonstrated in 15 of 16 patients by bDNA assay, 15 of 16 by CMV antigen assay, and 11 of 15 by CSF culture. When clinical criteria and results of the other two assays were used as reference standards, the sensitivity of bDNA was 94% and 100% and the specificity 95.2% and 100%; the CMV antigen assay sensitivity was 94% and 100% and specificity was 85.7% and 100%. Nine (90%) of 10 patients with polyradiculopathy and follow-up CSF culture showed a drop in CMV DNA after treatment; however, only 2 (20%) improved clinically. These results suggest that bDNA and antigen assays may be useful methods for the diagnosis of CMV polyradiculopathy, but treatment failures may not be due to inadequate antiviral activity.

A branched DNA signal amplification assay for quantification of nucleic acid targets below 100 molecules/ml.

The branched DNA hybridization assay has been improved by the inclusion of the novel nucleotides, isoC and isoG, in the amplification sequences to prevent non-specific hybridization. The novel isoC, isoG-containing amplification sequences have no detectable interaction with any natural DNA sequence. The control of non-specific hybridization in turn permits increased signal amplification. Addition of a 14 site preamplifier was found to increase the signal/noise ratio 8-fold. A set of 74 oligonucleotide probes was designed to the consensus HIV POL sequence. The detection limit of this new HIV branched DNA amplifier assay was approximately 50 molecules/ml. The assay was used to measure viral load in 87 plasma samples of HIV- infected patients on triple drug therapy whose RNA titers were <500 molecules/ml. In all 11 patients viral load eventually declined to below the detection limit with the new assay.

Increase in angiotensin II type 1 receptor expression immediately after ischemia-reperfusion in isolated rat hearts.

BACKGROUND: Myocardial ischemia is known to upregulate the systemic renin-angiotensin system, which influences myocardial ischemic events by affecting hemodynamics and hemostatic activity. This study was designed to examine whether angiotensin II (Ang II) receptor expression in the myocardium is altered immediately after ischemia-reperfusion. METHODS AND RESULTS: Isolated buffer-perfused Sprague-Dawley rat hearts were subjected to continuous perfusion (control, n=5) or to 25 minutes of global ischemia followed by 30 minutes of reperfusion (n=10). Autoradiographic analysis for Ang II receptors of multiple myocardial sections was performed. Whereas continuous perfusion of hearts resulted in minor changes in coronary perfusion pressure (CPP), left ventricular end-diastolic pressure (LVEDP), and developed left ventricular pressure (dLVP=LVSP-LVEDP), ischemia-reperfusion caused a marked increase in CPP and LVEDP and a decrease in dLVP, indicating severe cardiac dysfunction. Concurrently, total myocardial Ang II receptor expression was greater (P<.05) in hearts subjected to ischemia-reperfusion than in the continuously perfused control hearts. Most of the increase in Ang II receptor expression was due to an increase in type 1 receptor (AT1) expression (34.6+/-6.5 versus 18.2+/-4.4 fmol/g, P<.05), because Ang II type 2 receptor expression was unaffected. To examine the importance of AT1 receptor expression, another group of isolated rat hearts (n=5) was perfused with buffer containing losartan (10(-5) mol/L) and subjected to ischemia followed by reperfusion. Perfusion of hearts with losartan attenuated the ischemia-reperfusion-induced cardiac dysfunction. Perfusion of hearts with losartan also blocked the ischemia-reperfusion-induced increase in myocardial AT1 binding. CONCLUSIONS: These observations indicate that myocardial AT1 expression increases immediately after ischemia-reperfusion and contributes to cardiac dysfunction.