Identifying clinical risk factors correlate with suicide attempts in patients with first episode major depressive disorder.

BACKGROUND: Major depressive disorder (MDD) is the most common mental disorder associated with suicide attempts. When a patient first visits the clinic, clinicians are often expected to make concrete diagnose about acute suicidal risk. However, the timeliness of suicide attempts correlates with patients with MDD has not been tested. METHODS: We divided 1718 first-episode and untreated MDD outpatients into those who did not have suicide attempts (non-attempts), recent suicide attempters (≤14 days before assessment) and long - dated suicide attempters (> 30 days before assessment). Positive Symptom Scale of Positive and Negative Syndrome Scale (PANSS), the 17-item Hamilton Depression Scale, 14 - item Hamilton Anxiety Scale, and clinical global impression of severity scale (CGI-S) was assessed. Body mass index, some glycolipid metabolism and thyroid hormone parameters were measured. A gradient-boosted decision trees statistical model was used to generate equally weighted classification for distinguishing recent and long - dated suicide attempters from non-attempts. RESULTS: The classifier identified higher excitement, hostility, anxiety, depression symptoms and higher free thyroxine (FT4) as risk factors for recent suicide attempters with an estimated accuracy of 87% (sensitivity, 59.1%; specificity, 61.2 %). For long - dated suicide attempters' risk factors, single status, higher anxiety and hostility symptoms, higher LDLC and lower BMI, the estimated accuracy was 88% (sensitivity, 52.8%; specificity, 49.6%). CONCLUSIONS: Risk factors for suicide attempt among patients with MDD can be identified by integrating demographic, clinical, and biological variables as early as possible during the first time see a doctor.

Nested real-time quantitative polymerase chain reaction assay for detection of hepatitis B virus covalently closed circular DNA.

BACKGROUND: Successful treatment of hepatitis B can be achieved only if the template for hepatitis B virus (HBV) DNA replication, the covalently closed circular HBV DNA (cccDNA) can be completely cleared. To date, detecting cccDNA remains clinically challenging. The purpose of this study was to develop a nested real-time quantitative polymerase chain reaction (PCR) assay for detecting HBV cccDNA in peripheral blood mononuclear cells (PBMCs) and bone marrow mononuclear cells (MMNCs). METHODS: Based on the structural differences between HBV cccDNA and HBV relaxed circular DNA (rcDNA), two pairs of primers were synthesized as well as a downstream TaqMan probe. Blood and bone marrow samples were collected from hepatitis B patients and healthy controls. To remove rcDNA, samples were incubated with mung bean nuclease and the resultant purified HBV cccDNA was then amplified by nested real-time fluorescence quantitative PCR. The cccDNA levels were calculated using a positive standard. RESULTS: The nested real-time fluorescence quantitative PCR method for HBV cccDNA was successful, with a linear range of 3.0 × 10(2) copies/ml to 3.9 × 10(8) copies/ml. Of the 25 PBMC samples and 7 MMNC samples obtained from chronic hepatitis B or liver cirrhosis patients, 3 MMNC samples and 9 PBMC samples were positive for HBV cccDNA, while all of the 21 PBMC samples from healthy controls were negative. CONCLUSION: The nested real-time fluorescence quantitative PCR may be used as an important tool for detecting cccDNA in hepatitis B patients.

[A multicenter prospective randomized open comparative study on the treatment of ovulatory menorrhagia with tranexamic acid and norethisterone in China].

OBJECTIVE: To compare the efficacy and safety of tranexamic acid (TA) and norethisterone (NET) for the treatment of patients with ovulatory menorrhagia in China. METHODS: One hundred and thirty one patients with proven ovulatory menorrhagia from gynecologic clinics of 5 teaching hospitals located in 4 different cities in China were enrolled during Jul 2004 to Dec 2006. Among them 128 completed the study. Patients were randomly divided into two therapeutic regimen groups: TA 1 g thrice daily during menstrual cycle days (D) 1-5, 69 cases; or NET 5 mg twice daily on D19-26, 59 cases. The drugs were administered for 2 consecutive cycles, then withdrawn and patients were followed-up for 1 more cycle. Data on menstrual blood loss [estimated by pictorial blood assessment chart (PBAC)], length of menstrual periods, quality of life (QOL) evaluated by a 6 item health-related questionnaire were collected before, during each cycle and were compared. RESULTS: Both treatments led to significant decreases of mean PBAC scores and shorter duration of menstrual periods, and improved the QOL ranking during the two treatment cycles. The mean percentages of PBAC decrements in the TA first and second cycles were significantly greater than those in the NET corresponding cycles(35% vs 17% , P = 0.004; 44% vs 34%, P = 0.04 respectively). The success rate of TA second cycle was higher than that of the NET second cycle (41% vs 24%, P = 0.04). Improvement of QOL ranking in the TA first cycle was also significantly better than those in the NET first cycle (P = 0.03). The percentage of patients with at least 1 adverse event in TA group (19%) was significantly lower than that in NET group (35%, P = 0.04). Patients' willingness to continue the treatment in the TA second and follow-up cycles (94%, 79% respectively) were significantly higher than those in the corresponding cycles of NET groups (79%, 59% respectively; P = 0.01, P = 0.02). CONCLUSION: The regimen of TA 3 g daily during menstrual days 1-5 is a more effective and tolerable treatment than luteal phase norethisterone for patients with ovulatory menorrhagia.

[The pathogenetic role of peripheral T-lymphocytes activation-induced cell death in hemorrhagic fever with renal syndrome].

OBJECTIVE: To study the activation-induced cell death (AICD) in peripheral blood T-lymphocytes (PBL-T) from patients of hemorrhagic fever with renal syndrome (HFRS) and the effect of AICD on the pathogenesis of HFRS. METHODS: The PBL-T of patients with HFRS were isolated by negative selection with magnetic beads and cultured with or without phytohemagglutinin (PHA) and ionomycin. The apoptotic ratio of PBL-T was detected with TUNEL staining and assessed with flow cytometry. The level of caspase-3 of PBL-T in patients with HFRS was detected with Western blot. Characteristic gene fragment of Hantaan virus and Seoul virus was detected with reverse transcription polymerase chain reaction. RESULTS: In HFRS patients, the apoptotic ratio (27.79 +/- 0.99)% of PBL-T activated with PHA and ionomycin was significantly higher than that without activation (17.16 +/- 1.14)%, (P < 0.01). The apoptotic ratio of PBL-T in HFRS patients was higher than that of healthy control (P < 0.01), no matter with or without activation. The expression level of caspase-3 increased in the induced group of patients with HFRS and the expression level of 17 000 (the active component of caspase-3) increased significantly. The results of sequencing showed that amplification production from the serum are positive in 19 cases, seven being Hantaan virus and twelve Seoul virus. CONCLUSIONS: AICD in PBL-T was found in patients of with HFRS. The high expression rate of caspase-3 in the PBL-T of patients with HFRS was related with AICD in the patients with HFRS. In the HFRS patients with specific IgM antibody positive, characteristic fragments can be obtained with RT-PCR from 79.2% patients. AICD of T lymphocytes may play an important role in the pathogenesis of HFRS.

Role of activation-induced cell death in pathogenesis of patients with chronic hepatitis B.

AIM: To study and compare the difference of activation-induced cell death (AICD) in peripheral blood T-lymphocytes(PBL-Ts) from patients with chronic hepatitis B (CHB) and the normal people in vitro, and to explore the role of AICD in chronic hepatitis B virus (HBV) infection and the pathogenesis of CHB. METHODS: Twenty-five patients and fourteen healthy people were selected for isolation of PBL-Ts. During cultivation, anti-CD3 mAb, PMA and ionomycin were used for AICD of PBL-Ts. AICD ratio of PBL-Ts was detected with TdT-mediated dUTP nick end labeling and assessed by flow cytometry. RESULTS: When induced with anti-CD3, PMA and ionomycin in vitro, AICD ratio of PBL-Ts from CHB patients was significantly higher than that from healthy control (17.24+/-1.21 vs. 6.63+/-1.00, P<0.01) and that from CHB patients without induction (17.24+/-1.21 vs. 9.88+/-1.36, P<0.01). There was a similar AICD ratio of PBL-Ts between induction group and without induction group, but no difference was found before and after induction in healthy control. The density of INF-gamma in culture media of induction groups of CHB was lower than that of other groups (P<0.01). There was no difference between these groups in density of IL-10 (P>0.05). CONCLUSION: When induced during cultivation in vitro, PBL-Ts from CHB have AICD very commonly. This phenomenon has a potentially important relation with pathogenesis of CHB and chronicity of HBV infection.

[Hepatitis C virus nonstructural 5A protein inhibits tumor necrosis factor alpha mediated apoptosis of HepG2 cells].

OBJECTIVE: To study the suppressive effect of HCV nonstructural 5A (NS5A) protein on tumor necrosis factor alpha (TNF alpha) mediated apoptosis of HepG2 cells. METHODS: NS5A gene fragment was amplified by reverse transcription and nested polymerase chain reaction from serum samples positive for anti-HCV. The PCR product was directly cloned using TA cloning kit, and 2 independent clones were isolated, digested and sequenced. Then we constructed HCV NS5A expression plasmid (pcDNA3.1-NS5A), stably transfected into HepG2 cells with lipofectamine. Successful transfection of NS5A gene and expression of NS5A protein were confirmed by Western blot. Transfected cells were incubated with TNFalpha for 48 h, then labeled with Annexin V and visualized by fluorescence microscopy. To examine the effects of NS5A protein on the apoptotic signaling pathway, caspase-3 cleavages and release of cytochrome C were investigated in the transfectant treated with TNF alpha for 48 h and the cell cytosol was subjected to SDS-PAGE and Western blot analysis. RESULTS: The stable transfectant of HepG2 cells lines for HCV NS5A protein expression was achieved. The NS5A protein blocked the activation of caspase-3 and the release of cytochrome C in the TNF alpha treated cells. CONCLUSION: HCV NS5A protein inhibits TNF alpha mediated apoptosis of HepG2 cells in vitro.

Serum and ascites levels of macrophage migration inhibitory factor, TNF-alpha and IL-6 in patients with chronic virus hepatitis B and hepatitis cirrhosis.

OBJECTIVE: To study the potential role of macrophage migration inhibitory factor (MIF), tumor necrosis factor-alpha (TNF-alpha) and interleukin-6 (IL-6) in the development of chronic virus hepatitis B (CH) and hepatitis cirrhosis (HC). METHODS: The serum concentrations of MIF, TNF-alpha and IL-6 in 18 patients with chronic virus hepatitis B and in 14 patients with hepatitis cirrhosis without ascitic fluid, and the serum and ascites cytokine concentrations in 22 HC patients with ascitic fluid were detected by enzyme linked immunity sorbed assay. RESULTS: The cytokine concentrations of the patients were significantly higher than those of the controls. The serum levels of MIF, TNF-alpha and IL-6 of the 22 patients with ascitic fluid were higher than those of 14 HC patients without ascites. In the 18 patients with CH, the serum cytokine concentrations were the lowest. The serum cytokine concentrations of the 22 HC patients with ascites were significantly higher than those of the 14 HC patients without ascites (P<0.01). Their serum cytokine concentrations were significantly higher than those in the 18 patients with CH (P<0.01). The concentration of IL-6 in ascites was the highest among all the groups. The serum levels of MIF, TNF-alpha and IL-6 are correlated with alanine aminotransferase (ALT) in the patients with CH, but not in those with HC with or without ascites. CONCLUSIONS: These results indicated that MIF, TNF-alpha and IL-6 may participate in the pathological process of CH and cirrhosis, that IL-6 seems to play an important role in ascites formation, and that se-rum levels of MIF, TNF-alpha and IL-6 appear to reflect the severity of tissue injury in HBV disease.