Evaluation of mental status HIV-infected patients: implications for treatment.

OBJECTIVES: The aim of the study was to explore factors that may influence mental health status of HIV-infected patients, and to figure out a method that could effectively classify the patients by evaluating the severity of their mental health problems. METHODS: Eighty-five patients were recruited and divided into two groups: the low risk group (LRG, n = 38) and the high risk group (HRG, n = 47). All patients finished Symptom Check-List 90 Revised (SCL-90-R) which is a multidimensional questionnaire designed to screen for a broad range of psychological problems. RESULTS: SCL-90-R scores of HRG were significantly higher than those of the general population and did not differ from those of psychosomatic outpatients. Scores of LRG were significantly lower than those of psychosomatic outpatients and did not differ from those of the general population in most subscales. HIV-infected men having sex with men and unemployed patients had much higher incidence of mental health problems. CONCLUSION: Besides undistinguishable group psychotherapy, we call for a clinical screening by psychological questionnaires at the first step, and then at-risk or high-risk patients should be given corresponding and individualized treatments. More attention and health care should be given to HIV-infected men who have sex with men and unemployed patients.

The impact of highly active antiretroviral treatment on the blood profiles of patients with acquired immune deficiency syndrome.

This study retrospectively investigated the short- and long-term impact of highly active antiretroviral treatment (HAART) on the blood profiles of patients with acquired immune deficiency syndrome (AIDS), and their relationship with disease progression. CD4(+) T-cell count was measured by flow cytometry, plasma viral load of human immunodeficiency virus (HIV) RNA was detected by reverse transcription-polymerase chain reaction, and blood profile was determined by an automated analyser. CD4(+) T-cell count, total lymphocyte count (TLC) and haemoglobin concentration improved gradually in patients with AIDS after the initiation of HAART. Long-term effective HAART significantly increased CD4(+) T-cell counts TLC and haemoglobin concentrations, and reduced viral load to undetectable levels. An increase in haemoglobin was positively correlated with an increase in CD4(+) T-cells. These findings suggest that TLC is a valuable tool for determining the initiation of HAART, and that the haemoglobin concentration could be an additional indicator for long-term monitoring of HAART in resource-limited settings.

Novel monoclonal antibodies to ESAT-6 and CFP-10 antigens for ELISA-based diagnosis of pleural tuberculosis.

OBJECTIVE: To elucidate the potential of monoclonal antibodies (mAbs) of culture filtrate protein 10 (CFP-10) and early secretory antigenic target 6 (ESAT-6) in tuberculosis (TB) diagnosis. DESIGN: We generated and characterised monoclonal and polyclonal antibodies against Mycobacterium tuberculosis-specific antigens ESAT-6 and CFP-10 by immunising BALB/c mice with an ESAT-6/CFP-10 fusion protein. Stable hybridoma cell lines were established and mAbs were specifically identified by immunoblotting and immunoprecipitation. The mouse mAbs were used to coat plates, and biotin-labelled polyclonal antibodies were used to detect the antigens. One hundred and seventy-three samples of sputum culture supernatants and pleural effusion aspirates have been tested. RESULTS: The ESAT-6 enzyme-linked immunosorbent assay (ELISA) detected the culture supernatants and pleural effusion specimens that were positive for M. tuberculosis, but failed to identify M. tuberculosis-positive specimens in the non-M. tuberculosis culture supernatants or control specimens. This yielded a sensitivity of 95.4% and a specificity of 100% for the ESAT-6-specific ELISA. The CFP-10 ELISA presented less satisfactory sensitivity and specificity, of respectively 81.6% and 92.2%. Results showed positive detection rates of ESAT-6 and CFP-10 of 86.8% (33/38) and 76.3% (29/38) for the diagnosis of tuberculous pleural effusion in patients bacteriologically negative for M. tuberculosis culture. CONCLUSION: The ESAT-6 and CFP-10 ELISAs incorporating mAbs generated in this study serve as potential tools in the laboratory diagnosis of TB.

Membrane current-based mechanisms for excitability transitions in neurons of the rat mesencephalic trigeminal nuclei.

Since Hodgkin's first description of three classes of excitability in crustacean nerve axons (1948), theoretical studies have used mathematical models to demonstrate that small changes in the parameters describing ionic currents could result in transitions between classes of membrane excitability. However, these transitions have rarely been investigated experimentally. Here, we show that states of excitability in rat mesencephalic V (Mes V) neurons can be classified into three groups, with manipulations of the 4-aminopyridine sensitive K(+) current (I(4-AP)) or persistent Na(+) current (I(NaP)) leading to the corresponding transitions. However, alterations in the hyperpolarization-activated cation current (I(h)), tetraethylammonium (TEA)-sensitive K(+) current, or Cd(2+)-sensitive Ca(2+) current were ineffective in causing these transitions. These results provide experimental evidence for the excitability transitions predicted by Hodgkin and characterize their ionic mechanisms in Mes V neurons.

Construction of expression vector of hTERT- hIL18 fusion gene and induction of cytotoxic T lymphocyte response against hTERT.

AIM: Recombiant human telomerase reverse transcriptase (hTERT)-human IL-18(hIL18) was constructed to investigate its expression and biological function in eukaryotic cells. DCs transfected with hTERT-IL18 acquired stronger telomerase activity and were able to elicit an hTERT-specific cytotoxic T lymphocyte CTL response in vitro. METHODS: hIL-18 gene fragment was amplified by polymerase chain reaction (PCR) and TA cloned. The hIL-18 gene was then subcloned into eukaryotic expression vector pcDNA3.1(+) containing human TERT via a linker. The sequence of gene fusion was confirmed using both restrictive enzyme digestion and DNA sequencer. The expression vector with gene fusion was transfected into 3T3 cell line with Lipofectamine 2000. ELISA, Western blot, immunofluorescence stain were performed to determine the expression properties of hTERT-hIL18 in 3T3 cells. Its biological effect on the anti-apoptosis was measured by Flow cytometry and its effect on INF-gamma expression was determined using ELISA. After preparation of dendritic cells, hTERT-hIL18, hTERT, hIL-18 expression vectors were transfected into DCs respectively by electroporation to generate hTERT-specific DCs lines. The peripheral blood mononuclear cells PBMCs were stimulated with different DCs lines to create specific CTL. The response of target cell (leukemia cell line-K562 cell) to hTERT-specific CTL was evaluated by LDH release assay. RESULTS: The human IL-18 gene fragment was amplified from the human mononuclear cells and was inserted into pcDNA3.1(+)/hTERT vector successfully. The correct sequence was proved by both restrictive enzyme digestion and sequencing. The correct open reading frame was also verified. Fusion protein of hTERT-hIL18 was effectively expressed in eukaryotic cells, which was detected by both Western-blotting and immunofluorescence stain. The expressed recombinant fusion protein induced similar levels of INF-gamma to that of native IL-18 protein. FCM assay showed that the transfected fusion protein inhibited the apoptosis, which was consistent with the effects of hTERT as a universal tumor associated antigen. CTL assay shows that hTERT- hIL18 and hTERT gene-transfected DCs stimulated T-cell responses that recognized and lysed tumor target cells of high hTERT expression, whereas DCs transfected with hIL-18 gene didn't induced the response of tumor targets lyses. CONCLUSION: The Recombinant hTERT- hIL18 fusion protein had both biological activity of hTERT and hIL-18, indicating that this rationally designed protein can be further developed as novel tumor therapeutics. DCs transfected with hTERT-IL18 gene were capable of eliciting a stronger hTERT-specific CTL response in vitro.

A chemically modified carbon paste electrode with d-lactate dehydrogenase and alanine aminotranferase enzyme sequences for d-lactic acid analysis.

An amperometric biosensor was constructed for the analysis of d-lactic acid based on immobilizing d-lactate dehydrogenase(d-LDH), alanine aminotransferase (ALT), NAD(+), a redox polymer and polyethylenimine in carbon paste. The effect of addition of ALT in the paste, using enzyme sequences of ALT/d-LDH, was insignificant for d-lactic acid analysis. The responses of d-lactic acid in ALT/d-LDH paste electrode are the same as those in d-LDH paste electrode. However, the interference effect of pyruvate in the sample can be substantially reduced if sodium glutamate was applied in the carrier solution. When ALT immobilized in control porous glass as an immobilized enzyme reactor (IMER) was mounted in flow injection analysis system with the d-LDH paste electrode as detector for d-lactate analysis, the interference of the pyruvate can be significantly eliminated. The adverse effect of pyruvate in the samples for d-lactic acid analysis was reduced more effectively in ALT IMER with d-LDH electrode than in ALT/d-LDH electrode.