Clinical performance of quantitative PCR for the molecular identification of skeletal tuberculosis from formalin-fixed paraffin-embedded tissues.

BACKGROUND: At present, skeletal tuberculosis (TB) diagnosis is mostly by histopathology, but the positivity rate is low. There is a need to develop new methods for the molecular identification of this disorder. Therefore, we aimed to investigate the clinical utility of quantitative PCR (qPCR)-based diagnosis of skeletal TB from formalin-fixed paraffin-embedded (FFPE) tissues and its comparative evaluation with acid-fast bacillus staining (AFS). METHODS: We detected Mycobacterium tuberculosis (M. tuberculosis/MTB) DNA using qPCR and AFS in FFPE tissue samples from 129 patients suspected of having skeletal TB. The sensitivity, specificity as well as area under the curve (AUC) of qPCR and AFS were calculated. Meanwhile, some factors potentially affecting qPCR and AFS results were investigated. RESULTS: Overall, qPCR outperformed AFS in detecting M. tuberculosis. The AUC of qPCR was higher than that of AFS (0.744 vs.0.561, p < 0.001). Furthermore, decalcification of bone tissues did not affect the sensitivity and specificity of qPCR tests. Whereas it impacted the performance of AFS, decalcification increased AFS's specificity and decreased its sensitivity (p < 0.05). Moreover, qPCR had a significantly larger AUC than AFS in decalcified and non-decalcified groups (0.735/0.756 vs. 0.582/0.534, p < 0.001) respectively. Similarly, the AUC of PCR was more extensive than that of AFS regardless of skeletal TB patients with concomitant pulmonary TB or not (0.929 vs. 0.762; 0.688 vs. 0.524, p < 0.01). CONCLUSIONS: Our data demonstrate that qPCR offers superior accuracy for the detection of mycobacteria in FFPE tissues compared to traditional AFS, indicating its clinical value in osteoarticular TB diagnosis.

The novel HLA-C*01:255 allele, identified by Sanger dideoxy nucleotide sequencing in a Chinese individual.

HLA-C*01:255 differs from HLA-C*01:02:01:01 by one nucleotide in exon 2.

The novel HLA-B*54:01:12 allele, identified by Sanger dideoxy nucleotide sequencing in a Chinese individual.

HLA-B*54:01:12 differs from HLA-B*54:01:01:01 by one nucleotide in exon 2.

The novel HLA-B*38:103N allele, identified by Sanger dideoxy nucleotide sequencing in a Chinese individual.

HLA-B*38:103N differs from HLA-B*38:02:01:01 by one nucleotide in exon 3.

The novel HLA-DPA1*02:117 allele, identified by Sanger dideoxy nucleotide sequencing in a Chinese individual.

HLA-DPA1*02:117 differs from HLA-DPA1*02:02:02:01 by one nucleotide in exon 2.

The novel HLA-C*17:69 allele, identified by Sanger dideoxy nucleotide sequencing in a Chinese individual.

HLA-C*17:69 differs from HLA-C*17:01:01:02 by one nucleotide in exon 4.

The novel HLA-B*40:550 allele, identified by Sanger dideoxy nucleotide sequencing in a Chinese individual.

HLA-B*40:550 differs from HLA-B*40:01:02:01 by one nucleotide in exon 1.

The novel HLA-B*27:267 allele, identified by Sanger dideoxy nucleotide sequencing in a Chinese individual.

HLA-B*27:267 differs from HLA-B*27:04:01 by one nucleotide in exon 2.

The novel HLA-C*06:02:105 allele, identified by Sanger dideoxy nucleotide sequencing in a Chinese individual.

HLA-C*06:02:105 differs from HLA-C*06:02:01:01 by one nucleotide in exon 1.

Evaluation of human immunodeficiency virus type 1-infected Chinese patients treated with highly active antiretroviral therapy for two years.

The purpose of this study was to evaluate the long-term efficacy and safety of nevirapine in combination with didanosine and stavudine in the treatment of human immunodeficiency virus (HIV)1-infected Chinese patients in routine clinical practice. The study, from April 2003 to May 2005, with follow-up through 24 mo, was conducted at the Department of Infectious Diseases, Second Xiangya Hospital, Central-South University in Changsha, Hunan Province, China. Twenty-seven HIV1-infected patients received didanosine, stavudine, and nevirapine. Information from case notes regarding age, sex, side effects, viral load, naive and memory T cells, and CD4(+) and CD8(+) T cell count at baseline, 3, 6, 12, 18, and 24 mo was collected and analyzed. Virologic suppression, defined as an HIV RNA concentration of less than 50 copies/mL at months 3, 6, 12, 18, and 24, was considered the main outcome measure. Of 27 patients, 17 were men with a mean age 33.5 yr. The mean baseline viral load was 5.15 log copies/mL and the mean CD4(+) cell count was 185 cells/dL. Of 27 patients, 3 patients discontinued study medication; treatment was changed, because of side effects, from didanosine (ddI), stavudine (d4T), and nevirapine (NVP) to zidovudine, lamivudine, and NVP for 24 patients who had completed 24 mo of treatment with ddI, d4T, and NVP; and viral load suppression was attained in 17 patients (70.8%) at 12 mo, in 14 patients (58.3%) at 18 mo, and in 13 patients (56.6%) at 24 mo. The CD4 T cell count increased by 114 cells/microL (mean, 299 cells/microL) after 12 mo of treatment and by 132 cells/microL (mean, 317 cells/microL) after 24 mo of treatment. Naive T cells and memory cells also increased in number, but at a slower rate. Activated (CD38(+)) CD8(+) T cells were elevated at baseline (67.7%) and declined by month 24 (49.7%), but did not reach normal levels. We conclude that a regimen of NVP with ddI and d4T provided durable suppression of plasma viral load in HIV-infected patients, with significant improvement in the CD4 cell count, and can be well tolerated by patients with HIV-1 infection.

The complete mitochondrial genome of Luehdorfia chinensis Leech (Lepidoptera: Papilionidae) from China.

The complete mitochondrial genome of Luehdorfia chinensis that is endemic to China is determined. The circular genome is 15 550 bp in length and comprises 13 protein-coding genes (PCGs), 22 tRNAs, two rRNA gene and a control region. Gene order is identical to that of the putative ancestral arrangement of insects. The nucleotide composition of heavy strand is A (40.3%), C (11.2%), T (41.1%) and G (7.3%). All PCGs start with a typical ATN codon except for the gene COI that uses AAC as the start codon. Bayesian analyses support the monophyly of genus Papilio and the sister relationship of Luehdorfia chinensis and L. taibai.

The preliminary efficacy of interferon-alpha and ribavirin combination treatment of chronic hepatitis C in HIV-infected patients.

BACKGROUND: It is internationally accepted that in drug-naïve individuals with hepatitis C virus (HCV) and human immunodeficiency virus (HIV) co-infection, chronic hepatitis C should be treated first if the CD4 cell count does not require the initiation of anti-retroviral therapy. Present paper evaluated the clinical effect and side-effect of interferon-alpha (IFN-alpha) and ribavirin (RBV) combination therapy for Chinese patients with HCV-HIV co-infection, and compared with them for HIV infection alone. METHODS: Ten patients with HCV-HIV and 17 patients with HCV received 5 million unit IFNalpha-2b every other day intramuscularly, and 300 mg RBV triple daily by oral. Dynamic observations were made for HCV RNA and HIV RNA loads, CD4+ and CD8+ T lymphocyte counts, liver function and blood cell measurement, and the medicine side-effects. RESULTS: After 12-week and 24-week treatments of IFN-alpha and RBV combination therapy, mean HCV RNA levels reduced 1.14 logs and 1.56 logs from the baseline at week 0 in HCV-HIV co-infection, and reduced 1.48 logs and 1.75 logs in HCV infection, respectively. The HIV RNA levels decreased 1.22 logs and 1.32 logs from the base line; however, there were no obvious different changes at T lymphocyte counts of HCV-HIV and HCV patients through 24-week treatments. Whole 27 patients showed satisfactory biochemical response to therapy. There were some mild or mediate influence-like symptoms, intestinal uncomfortable and depressed blood cell counts in early stage of the treatments. No neuropsychiatric and auto-immune disorders were found. CONCLUSIONS: IFN-alpha and RBV combination therapy had similar anti-HCV effects during 24-week treatment for HCV-HIV and HCV infected Chinese patients, and some anti-HIV effect. There were no obvious different biochemical responses and side-effects between two groups above.

[Interferon-alpha and ribavirin combination therapy for co-infection of hepatitis C virus and human immunodeficiency virus].

OBJECTIVE: To evaluate the clinical effect and side-effect of interferon-alpha (IFN-a) and ribavirin (RBV) combination therapy for Chinese patients with co-infection of hepatitis C virus (HCV) and human immunodeficiency virus (HIV), and to compare them with only HIV infection patients. METHODS: 10 patients with HCV-HIV and 17 patients with only HCV infection received 5 million units of IFNalpha-2b every other day intramuscularly, and 300 mg RBV orally three times a day. Dynamic observations were done for HCV RNA and HIV RNA loads, CD4+ and CD8+ T lymphocyte counts, liver function and blood cell measures, and the side-effects of the medicines. RESULTS: After 12 weeks and 24 weeks of IFNalpha and RBV combination therapy, mean HCV RNA levels reduced 1.14 log (t = 3.843, P < 0.01) and 2.08 log (t =6.564, P < 0.01) from the baseline at week 0 in the HCV-HIV co-infection group, and reduced 1.48 log (t = 6.438, P less than 0.01) and 2.33 log (t = 7.343, P < 0.01) in the HCV infection group. Meanwhile, the HIV RNA levels decreased 1.22 log (t = 3.662, P < 0.01) and 1.73 log (t = 6.119, P < 0.01) from the base line. However, there were no obvious different changes among T lymphocyte counts of HCV-HIV and HCV patients at week 0, week 12 and week 24. All 27 patients showed satisfactory biochemical response to therapy. There were some mild or moderate influenza-like symptoms, intestinal discomfort and decreased blood cell counts in the early stages of the treatments. No neuropsychic and auto-immune disorders were found. CONCLUSIONS: IFNalpha-2b and RBV combination therapy showed similar anti-HCV effects during the 24 week treatment for HCV-HIV and HCV infected patients, and some anti-HIV effect was also observed. No obvious different biochemical responses and side-effects were found between the above two groups.

Dietary fiber intake and risk of renal cell carcinoma: evidence from a meta-analysis.

The aim of this study was to investigate the possible relationships between dietary fiber intake and risk of renal cell carcinoma (RCC). Electronic databases including MEDLINE, EMBASE and Web of Science were searched to find eligible studies. Random-effects relative risk (RR) and its corresponding 95 % confidence interval (CI) were used. Besides, random-effects dose-response analyses were also performed to clarify the dose-response relations. Finally, publication bias was assessed by Egger's test and Begg's test. All p values were two tailed. Seven studies, including two cohort studies and five case-control studies, were eligible and included in this meta-analysis. Overall analysis in highest versus lowest level revealed that total dietary fiber intake was associated with reduced RCC risk (RR 0.84, 95 % CI 0.74-0.96). In addition, pooled estimated data showed that risk of RCC was significantly associated with vegetable and legume fiber intake (RR 0.70, RR 0.80, respectively), but not with fruit and cereal fiber intake (RR 0.92, RR 1.04, respectively). However, in dose-response analysis, no significant association was reported. Finally, no publication bias was detected by Egger's or Begg's test. The dietary fiber intake, especially vegetable and legume fiber, may be associated with reduced RCC risk. Considering the limitations of the included studies, more well-designed prospective studies will be needed to confirm our findings.