[Caspase1 Inhibitor Ac-YVAD-CMK Prevents and Treats the Acute Graft Versus Host Disease in Mice].

OBJECTIVE: To explore the effect of Caspase 1 inhibitor Ac-YVAD-CMK on acute graft-versus-host disease (aGVHD) after allogeneic hematopoietic stem cell transplantation(allo-HSCT) and its mechanism. METHODS: Experiments were divided randomly into 3 groups: allogeneic hematopoietic stem cell transplantation combined with splenic cell infusion group (TS group, n=12), allogeneic hematopoietic stem cell transplantation combined with splenic cell infusion plus injection of low dose Caspase 1 inhibitor group (TS+low dose of C group, n=16) and plus high dose Caspase1 inhibitor (TS+high dose of C group, n=19). The body weight of mice in each group was dynamically detected, and the clinical manifestation of GVHD and score of aGVHD were determined, and the chimerism rate of mice was detected after transplantation for 14 days. Th1, Th2 and Th17 cells in peripheral blood were examined by flow cytometry. Peripheral proinflammatory cytokines IL-1ß, IFN-γ, IL-1α and IL-18 were examined by enzyme-linked immunosorbent assay(ELISA). The tissues sections of GVHD target organs (liver, lung, colon and skin) were stained with HE for histopathologic examination and histopathologic score. RESULTS: Ac-YVAD-CMK could alleviate murine aGVHD and pathological injury, decreare the incidence and severity of aGVHD in recipient mice. The detection of Th cell subsets in peripheral blood by flow cytometry showed that compared with TS group, the Th1 cell ratio in TS+low dose of C and TS+high dose of C groups was significantly reduced (P<0.05), while the Th2 and Th17 cell ratio was significantly enhanced (P<0.05) in TS+low dose of high dose of C groups. The detection of peripheral inflamematory cytokines by ELISA demonstrated that the inflammatory cytokines including IL-1ß,IFN-γ,IL-18 and IL-1α were reduced significantly (P<0.05). CONCLUSION: Ac-YVAD-CMK can improve aGVHD by inhibiting Caspase 1 and reducing the release of some inflammatory cytokines, thereby alleviated the aGVHD pathological damage.

[Clinical study of standard dose of rituximab for the treatment of refractory primary immune thrombocytopenia].

OBJECTIVE: To evaluate the efficacy and safety of rituximab on B-lymphocytes and anti-platelet glycoprotein-specific antibodies in patients with refractory primary immune thrombocytopenic (ITP). METHODS: Thirty-one ITP patients with a median age of 36 years (range 16 - 56 years) received solely intravenous rituximab at the dose of 375 mg/m(2) once weekly for consecutive 4 weeks. Lab studies included complete blood count, serum concentrations of IgG, IgM and IgA. CD3(+), CD4(+), CD8(+), CD19(+) and CD20(+) cell numbers were assayed by flow cytometry and anti-platelet glycoprotein-specific antibodies (GPIIb/IIIa, GPIb/IX) were assayed by monoclonal antibody-specific immobilisation of platelet antigens (MAIPA) prior to and following rituximab therapy. The response was evaluated according to the response criteria of international working group of ITP. RESULTS: Complete responses were achieved in 12 cases, response in 7 cases, and no response in 12 cases. Responses were sustained 2 to 28 months (median 6 months) with 4 cases relapsed. After 4 weeks of rituximab therapy, GPIIb/IIIa and GPIb/IX disappeared in responded patients, and CD 19(+)/CD20(+) cells were almost depleted in all patients. As expected, the serum concentrations of IgG, IgM, IgA, and the T cell counts were not changed after therapy. Four patients developed infusion-related reaction, 1 impaired renal function, and 3 secondary infections. CONCLUSION: Rituximab is effective and safe, and the adverse reaction is tolerable.

[Analysis of liver function index and clinical characters in 122 patients with acute hepatitis E virus coinfection with other hepatitis virus].

OBJECTIVE: To compare the liver function index and clinical characters in 122 patients with acute hepatitis E virus overlapping with other infection. METHODS: The liver function index and clinical characters of 122 patients with acute hepatitis E virus overlapping infection and 40 patients with acute hepatitis E were retrospectively analyzed. RESULTS: No significant differences of ALT, AST, TBIL, DBIL were found between acute hepatitis E groups and overlapping infection hepatitis A or hepatitis B (P > 0.05). However, there were significant differences of Albumin (ALB) and Globulin (GLO) were found between acute hepatitis E groups and overlapping infection hepatitis B (P < 0.01). In acute hepatitis E overlapping infected hepatitis B or hepatitis A patients, more and severe complications were also observed. CONCLUSION: The patients with acute Hepatitis E virus, especially Hepatitis E virus overlapping infection, need to pay more clinical monitor, prevent complication early and lower death rates.

[Analysis of SCCmec genotyping and antimicrobial susceptibility tests in methicillin resistant Staphylococcus aureus].

OBJECTIVE: To investigate the SCCmec genotyping, subtype and antimicrobial susceptibility tests in methicillin resistant staphylococcus aureus to guide the clinical treatment and provide the proof for molecular epidemiology. METHODS: To detect mecA gene and SCCmec genetyping and subtype in 50 MRSA by PCR. According to CLSI's guideline, antimicrobial susceptibility tests were performed with disk diffusion. RESULTS: All 50 MRSA had mecA genes. 45 strains were SCCmec III types; 3 strains were SCCmec III A types; 2 strains were SCCmec II types. There were no SCCmec I and SCCmec IV types. SCCmec II, SCCmec III and SCCmec III A type strains were all multiresistant. CONCLUSION: 50 MRSA are all multiresistant. SCCmec III are the main types.

[Detection of alpha-fetoprotein-L3 using agglutinin-coupled spin column to be used in diagnosis of hepatocellular carcinoma].

OBJECTIVE: To explore the effect of use of lens culinaris agglutinin (LCA)-coupled spin column (ACSC) in detection of alpha-fetoprotein (AFP) isoform AFP-L3 and to evaluate the value of AFP-L3 as a biomarker in diagnosis of hepatocellular-carcinoma (HCC). METHODS: The serum samples of 132 patients with elevated AFP level (20-1000 microg/L), 79 diagnosed as with HCC and 53 with benign liver diseases (35 with liver cirrhosis and 18 with chronic hepatitis) underwent ACSC to isolate the fraction of AFP-L3. The contents of AFP and AFP-L3 were detected by micro-particle immunoassay. The ratio of AFP-L3 to total AFP, AFP-L3%, was calculated. Correlation between the abnormally elevated AFP-L3% and HCC was analyzed. RESULTS: Detection of AFP-L3% using ACSC method was operating friendly. The average value of AFP-L3% in the patients with HCC was 36.4%, significantly higher than those of the patients with benign liver diseases (5.3% respectively, P < 0.01). The area under the receiver operating characteristic (ROC) curve of AFP-L3% was 0.807. Taking AFP-L3% > or = 10% as diagnostic criteria, the sensitivity of AFP-L3% in HCC diagnosis was 84.8% (67/79) and the specificity was 92.5% (49/53), with a total conformity rate of 87.9% compared to the confirmed clinical diagnosis. Conclusion ACSC is of clinical value in detecting AFP-L3. AFP-L3% is a valuable biomarker in diagnosis and prediction of prognosis of HCC.

A proton-activated, outwardly rectifying chloride channel in human umbilical vein endothelial cells.

Extracellular acidic pH-activated chloride channel I(Cl, acid), has been characterized in HEK 293 cells and mammalian cardiac myocytes. This study was designed to characterize I(Cl,acid) in human umbilical vein endothelial cells(HUVECs). The activation and deactivation of the current rapidly and repeatedly follows the change of the extracellular solution at pH 4.3, with the threshold pH 5.3. In addition, at very positive potentials, the current displays a time-dependent facilitation. pH-response relationship for I(Cl,acid) revealed that EC(50) is pH 4.764 with a threshold pH value of pH 5.3 and nH of 14.545. The current can be blocked by the Cl(-) channel inhibitor DIDS (100 microM). In summary, for the first time we report the presence of proton-activated, outwardly rectifying chloride channel in HUVECs. Because an acidic environment can develop in local myocardium under pathological conditions such as myocardial ischemia, I(Cl,acid) would play a role in regulation of EC function under these pathological conditions.

[A clinical analysis of HBV reactivation in patients with malignant tumors].

OBJECTIVE: To observe the anti-viral therapy effect on HBV reactivation in malignant tumor patients and hepatitis B virus carriers after their cancer chemotherapy. METHODS: Thirteen cancer patients but also chronic hepatitis B virus carriers were enrolled in this study. They were randomly put into two groups. Eight patients were put in the therapeutic group. They all had abnormal liver functions induced by the reactivation of HBV after their cancer chemotherapy. Then they were treated with lamivudine. The other 5 cases were treated with lamivudine before their cancer chemotherapy when their serum HBV DNA levels were less than 10(3) copies/ml (preventive therapeutic group). The two groups were followed-up with liver function tests and serum HBV DNA level measurements. RESULTS: Among the 8 cases of the therapeutic group, 5 cases died of liver failure; cancer chemotherapy was postponed or even terminated in 3 patients due to liver function abnormality and anti-virus treatment was started. In the preventive therapy group, no HBV reactivation was observed in any of the 5 cases. CONCLUSION: For HBV carrier cancer patients, an anti-viral therapy before their cancer chemotherapy seems to be very important.

[The diagnostic value of platelet glycoprotein-specific autoantibody detection in idiopathic thrombocytopenic purpura].

To detect levels of platelet glycoprotein-specific autoantibody in idiopathic thrombocytopenic purpura (ITP), chronic aplastic anemia (CAA), hematologic malignancies and healthy volunteers, and evaluate the clinical significance of platelet glycoprotein-specific autoantibody level in diagnosis for ITP, anti-GPIb/IX, anti-GPIIb/IIIa, anti-GPIV and anti-GPV auto-antibodies were detected contemporaneously by a modified monoclonal antibody immobilization of platelet antigen assay (modified MAIPA). The results showed that the total positive rate of antibodies against platelet GPIb/IX, GPIIb/IIIa, GPIV, GPV were 69.99%, 10%, 20% and 0% in ITP, CAA, hematologic malignancy group and healthy volunteers respectively. There was significant difference between ITP and CAA (chi(2) = 20.71, P < 0.005), between ITP and hematologic malignancy group (chi(2) = 12.22, P < 0. 005). There was no positive finding in the healthy control. It is concluded that platelet glycoprotein-specific autoantibody has high value for the diagnosis of ITP,many kinds of antibodies detection at one time can enhance sensitivity, MAIPA is a specific assay for the diagnosis of idiopathic thrombocytopenic purpura.

[The relationship among the counts of platelet, thrombopoietin and spleen index in patients with liver cirrhosis].

OBJECTIVE: To determine the reason of thrombocytopenia in patients with liver cirrhosis, we studied the relationship among platelet counts, serum thrombopoietin (TPO) level and spleen index. METHODS: Serum TPO, platelet counts and spleen index were measured in 71 cirrhotic patients. TPO was measured with ELISA method, spleen index were measured on ultrasonography by the same doctor. RESULTS: Platelet counts in patients with cirrhosis were lower than that of healthy group [(109.20+/-53.39) vs (169.63+/-26.60) x 10(12)/L, P<0.05]. Serum thrombopoietin level in patients with cirrhosis was similar to that of healthy group [(436.42+/-258.97) vs (412.63+/-132.80) pg/ml, P>0.05]. However, serum thrombopoietin level decreased as liver disease aggravated, [(526.13+/-317.44) pg/ml in Child-Pugh grade A, (445.22+/-214.90) pg/ml in grade B and (311.45+/-182.66) pg/ml in grade C, grade A vs. Grade C, P<0.05]. However, decline in platelet counts was accompanied with incline in spleen index coordinately. 35 of 71 cirrhotic patients had normal platelet counts whereas 36 of them had thrombocytopenia. Thrombopoietin levels were higher in non-thrombocytopenia group than in thrombocytopenia group [(529.43+/-282.64) vs. (351.27+/-228.25)pg/ml, P<0.01]; but spleen index of two groups showed no difference [(29.65+/-12.00) vs. (36.35+/-12.68) cm2, P>0.05]. Correlation was found between thrombopoietin level and platelet counts (r=0.252, P=0.025); no correlation was found between spleen index and platelet counts (r=-0.238, P=0.062). CONCLUSION: The decline serum TPO levels might play an important role for thrombocytopenia in patients with liver cirrhosis.

[The contrast study of Pre-S1 protein, HBV-DNA and HBeAg in diagnosing viral replication in patients with chronic hepatitis B].

OBJECTIVE: To determine the role of Pre-S1 protein in diagnosing viral replication in patients with chronic hepatitis B. METHODS: 104 consecutive patients with chronic hepatitis B were included in the study, liver biopsy were performed in all patients. Serial serum samples were studied with the quantitative determination of HBV-DNA by a quantitative PCR assay, determination of Pre-S1 protein by ELISA. RESULTS: The positive rates of HBV-DNA and Pre-S1 protein in patients with HBsAg HBeAg anti-HBc (+) both were 96.5%. The positive rates of HBV-DNA and Pre-S1 protein in patients with HBsAg anti-HBe anti-HBc (+) were 81.5%, 72.3%, respectively. The positive rates of HBV-DNA and Pre-S1 protein in patients with HBsAg anti-HBc (+) were 87.5%, 75.0%, respectively. It represented some patients with HBeAg (-) anti-HBe (+/-) still had viral replication. HBV-DNA>10(3) copy/ml as positive criteria for diagnosing viral replication, the positive rate of HBeAg, Pre-S1 were 31.5% (28/89), 80.9% (72/89) in patients with HBV-DNA>10(3) copy/ml, respectively. The concordance rates of HBeAg, Pre-S1 with HBV-DNA were 40.0% (42/104), 82.0% (85/104), respectively. CONCLUSION: It showed that Pre-S1 was more sensitive than HBeAg in diagnosing viral replication in patients with chronic hepatitis B.

[Preliminary report on primary drilling procedure at the time of hydroxyapatite orbital implant].

OBJECTIVE: To evaluate the clinical effect of the drilling and inserting a motility peg at the time of hydroxyapatite orbital implant. METHODS: After drilling and insertion of a titanium motility peg, a hydroxyapatite orbital implant was inserted in 31 consecutive patients. Of the 31 patients, 23 had been referred for primary enucleation and 8 for secondary implant surgery. The titanium-threaded sleeve was driven into the hydroxyapatite after which the titanium flat-headed peg was inserted into the sleeve before orbital implantation. The flat-headed peg was replaced with a titanium motility/support peg 3 to 7 months later. RESULTS: Follow-up ranged from 3.0 to 11.0 months (mean, 6.9 months). All patients were successfully fit with prostheses. Prosthetic motility was acceptable in each patient. 12 of 31 titanium flat-headed pegs became spontaneously and partially exposed within 3 to 7 months. There were no cases of secondary infection, implant exposure or extrusion of the motility peg. CONCLUSION: The hydroxyapatite orbital implant primary drilling procedure gives good cosmetic results with good motility of the artificial eye. It does not result in increased complications, and is an effective and efficient surgical option.