Objective:To investigate the role of serum inflammatory cytokines in the development of acute coronary syndrome (ACS). Methods: All of enrolled patients were diagnosed by clinical and coronary angiographic features and divided into four groups, the acute myocardial infarction (AMI) group, unstable angina pectoris (UAP) group, stable angina pectoris (SAP) group and control group. The values of high-sersitivity C-reactive protein(hs-CRP), matrix metallopeptidase 9(MMP-9) and tumor necrosis factor-a (TNF-a) in serum were measured by cytokine detection equipment system (B10-RAD Biological Technology Co.Ltd, USA) and analysed in four groups with statistics. Results: Compared with SAP and the control groups, the levels of TNF-a and MMP-9 were increased significantly in AMI group(P <0.01). The level of serum hs-CRP was significantly higher in AMI group than that in UAP, SAP and control groups (P < 0.05). There were no differences in the levels of hs-CRP and MMP-9 between UAP, SAP and control groups (P> 0.05). It was found that there was positive relation between hs-CRP, MMP-9 and TNF-a by Pearson correlation analysis. Conclusion:There was obvious relation between coronary heart disease and inflammation. The cytokines characterized by the increases of hs-CRP, TNF-a and MMP-9 were involved in the formation and progression of atherosclerosis and served as markers of unstable plagues.

Objective To investigate the prognostic factors of cervical high-grade squamous intraepithelial lesions treated by cold knife conization with negative margin.Methods Two hundred and sixty-six women with cervical high-grade squamous intraepithelial lesions treated by cold-knife conization with negative margins at Beijing Hospital between Jan 1999 and Jan 2004 were analyzed retrospectively.All patients were followed up with cytology,high-risk human papillomavirus(HPV)test and eolposcopy if necessary.Results The cervical CIN recurrence rate was 8.6% with no incidence of invasive cervical cancer after a median follow-up of 46 months.The recurrence was related to the grade of lesions and gland involvement pathologically.One of 20(5.0%)cases with cervical intraepithelial neoplasia(CIN)Ⅱ,9 of 164(5.5%)cases with CIN Ⅲ(excluding carcinoma in situ,CIS)and 13 of 82(15.8%)cases with CIS recurred(P

OBJECTIVETo study the relationship between the IFN-gamma producing cell specific for recipient (IFN-gamma-PCSR) in allogeneic stem cell transplantation (allo-HSCT) and acute graft versus host disease (aGVHD).

METHODSIn 37 consecutive HLA-identical sibling allo-HSCT pairs, peripheral blood mononuclear cells (PBMNC) from donors before allo-HSCT and recipients after allo-HSCT were taken as responder cells (RC), and PBMNC from recipients before allo-HSCT as allogeneic stimulator cells (allo-SC) in mixed lymphocyte reaction (MLR). IFN-gamma-PCSR in PBMNC were assayed using enzyme linked immunospot assay (ELISPOT).

RESULTSPretransplantation frequencies of IFN-gamma-PCSR in donor PBMNC were significantly higher in aGVHD group than in non-aGVHD group (P < 0.01) and IFN-gamma PCSRs (>or= 20/2 x 10(5)RC) were significantly associated with the occurrence of grade II-IV GVHD (P < 0.05). Compared with that before allo-HSCT, IFN-gamma PCSR in PBMNC of aGVHD patients was significantly increased (P < 0.05). When PBMNC from aGVHD patients reacted with donor PBMNC, the IFN-gamma PC was significantly lower than that with recipient PBMNC before allo-HSCT. Longitudinal analysis of IFN-gamma PCSR following allo-HSCT showed that compared with that in patients without aGVHD, the IFN-gamma PCSR were significantly higher in patients with that in aGVHD on day +14 (P < 0.01) and day +28 (P < 0.01), respectively. After immunosuppressive therapy for 7 days, IFN-gamma PC declined significantly (P < 0.05).

CONCLUSIONThe recipient-specific IFN-gamma PC is closely correlated with the allo-response during allo-HSCT and may be helpful for the prediction, diagnosis and monitoring of aGVHD.

Adolescent ; Adult ; Female ; Graft vs Host Disease ; immunology ; Hematopoietic Stem Cell Transplantation ; Humans ; Interferon-gamma ; biosynthesis ; immunology ; Lymphocyte Culture Test, Mixed ; Male ; Middle Aged ; T-Lymphocytes ; immunology ; metabolism ; Transplantation, Homologous ; immunology ; Young Adult

In order to explore a new way to study allogeneic reactive T lymphocytes, detection of cytokine-secreting T lymphocytes after allogeneic peripheral blood mononuclear cells (PBMNCs) stimulation and investigation of its clinical significance were performed. A novel cytokine secretion assay (CKSA) was first applied to detect T lymphocytes secreting cytokine including IFN-gamma, IL-4 and IL-10 at single cell level in human mixed lymphocytes reaction. IFN-gamma-secreting T cells from PBMNCs were then evaluated in 2 patients with acute graft versus host disease (aGVHD) after allogeneic bone marrow transplantation. The results showed that compared with IL-4 and IL-10 (which were 0.12 +/- 0.03% and 0.10 +/- 0.03% respectively), a sizable proportion of IFN-gamma-secreting T lymphocytes could be detected (1.12 +/- 0.13)% after allogeneic PBMNCs stimulation. Preliminary results indicated that frequency of IFN-gamma-secreting T lymphocytes correlated with the onset and severity of clinical aGVHD. In conclusion, it is feasible to detect IFN-gamma secreting T lymphocytes after allogeneic PBMNCs stimulation and to apply the CKSA technique for clinical identification of aGVHD.
Acute Disease ; Cytokines ; secretion ; Graft vs Host Disease ; etiology ; Humans ; Interferon-gamma ; secretion ; Interleukin-10 ; secretion ; Interleukin-4 ; secretion ; T-Lymphocytes ; immunology ; Transplantation, Homologous ; immunology

OBJECTIVETo detect, enrich and expand the cytokine secreting T lymphocytes after allogeneic PBMNCs stimulation.

METHODSThe novel cytokine secretion assay (CKSA) was applied to detect T lymphocytes secreting IFN-gamma at single cell level in human mixed lymphocytes reaction. IFN-gamma secreting T cells were enriched by means of magnetic sorting system and expanded with OKT(3), anti-CD(3)mAb and IL-2 combination. Antigen specificity of the expanded cells was confirmed using enzyme linked immunospot assay.

RESULTSA sizable proportion of IFN-gamma secreting T lymphocytes could be detected [(1.12 +/-0.13)% compared with (0.23 +/-0.07)%] and be further enriched to (67.3 +/-10.5)%, or (93.8 +/-22.1) fold. T lymphocytes could be expanded up to 600-fold within 21-28 days and the specific IFN-gamma response of expanded cells was confirmed with stimulation of the relevant allogeneic PBMNC, which was significantly higher than the irrelevant PBMNC control.

CONCLUSIONIt is feasible to detect significantly increased IFN-gamma secreting T lymphocytes after allogeneic PBMNCs stimulation based on the CKSA technique at single cell level and these cells can be efficiently enriched and expanded for further research.

Antibodies, Monoclonal ; pharmacology ; CD28 Antigens ; immunology ; Cell Proliferation ; Cells, Cultured ; Cytokines ; secretion ; Graft vs Host Disease ; immunology ; Humans ; Interferon-gamma ; secretion ; Interleukin-2 ; secretion ; Leukocytes, Mononuclear ; cytology ; immunology ; Lymphocyte Culture Test, Mixed ; Muromonab-CD3 ; pharmacology ; T-Lymphocytes ; cytology ; immunology

To investigate the mechanism and its significance of the depression of telomerase activity in HL-60 cells exposed to homoharringtonine (HHT), the semi-quantitative RT-PCR and PCR-ELISA were used to detect the expression of hTERT mRNA and telomerase activity in HL-60 cells, the apoptotic rate of HL-60 cells was assayed with TUNEL. The results indicated that compared with the control, the level of hTERT mRNA synthesis in HL-60 cells treated with HHT at 0.005 - 0.03 micro g/ml for 12 hours did not change significantly, but it reduced obviously at 0.04 micro g/ml and was undetectable at 0.05 micro g/ml. When HL-60 cells incubated with HHT at 0.02 micro g/ml for various hours, the expression of hTERT mRNA did not decrease after 6 - 18 hours and reduced significantly after 24 hours and was undetectable at 30 hours. The tendency for suppression of telomerase activity was consistent with decrease of the expression of hTERT mRNA in HHT-treated HL-60 cells. The apoptotic rate of HL-60 cells was apparently increased with the depression of hTERT mRNA and telomerase activity. In conclusion, the transcription of hTERT mRNA in HL-60 cells can be inhibited by HHT. The relationship between regulation of hTERT mRNA expression and HHT-induced apoptosis is worth investigating furtherly.
Apoptosis ; drug effects ; DNA-Binding Proteins ; Dose-Response Relationship, Drug ; HL-60 Cells ; Harringtonines ; pharmacology ; Humans ; RNA, Messenger ; analysis ; Telomerase ; genetics ; metabolism

OBJECTIVETo explore the effect of Sedum sarmentosum Bunge Extract (SSBE) on severe acute pancreatitis (SAP) induced acute lung injury (ALI) model rats and their excessive inflammatory reactions.

METHODSForty-two healthy adult male Sprague-Dawley (SD) rats were randomly divided into 3 groups, the sham-operated control group (C), the SAP group (SAP), and the SSBE treated group (SSBE), 14 in each group. SAP induced ALl rat model was induced by retrograde injection of 5% sodium taurocholate (1 mL/kg) into the pancreatic duct. SSBE (100 m/kg) was administrated subcutaneously after the establishment of the SAP model. Equal dose of SSBE was injected again 12 h later. Equal volume of normal saline was administrated in the same way for rats in the C group and the SAP group. Rats were sacrificed after successful modeling and samples taken at 12 and 24 h. Pathological changes in the pancreas and the lung tissue were observed under light microscope. The ascites, serum amylase (AMS), wet/dry proportion (W/D) of the lung tissue, activities of myeloperoxidase (MPO), interleukin-1 (IL-1), IL-6, and tumor necrosis factor-alpha (TNF-alpha) were also measured.

RESULTSAscites and serum AMS activities significantly increased; MPO, IL-1, IL-6, TNF-alpha contents, and W/D ratio also significantly increased in the SAP group, when compared with the C group (P<0.05). Compared with the SAP group, those parameters were all attenuated in the SSBE group at 12 and 24 h (P<0.05, P<0.01). Pathological changes in the pancreas and the lung tissue were alleviated in the SSBE group under light microscope. The injury degree ranged between that of the C group and the SAP group.

CONCLUSIONSSBE could relieve the ALl in SAP model rats, which could be achieved through alleviating inflammation responses of SAP rats.

Acute Lung Injury ; drug therapy ; etiology ; Animals ; Drugs, Chinese Herbal ; therapeutic use ; Interleukin-1 ; Interleukin-6 ; Lung ; Male ; Pancreas ; Pancreatitis ; complications ; drug therapy ; Peroxidase ; Rats ; Rats, Sprague-Dawley ; Sedum ; Taurocholic Acid ; Tumor Necrosis Factor-alpha

Objective To establish a pre -column derivatization and HPLC method for the determination of free amino acids in serum and urine with phenyl isothiocyanate ( PITC ) as derivatization reagent . Methods The determination was performed on waters e 2695 sys-tem.Sample was separated by Shim -pack VP-ODS column(250 mm × 4.6 mm,5 μm ) with a gradient elution of mobile phases composed of methanol/acetate/H2 O( pH 6.5 ) , the content of amino acid were tested by UV at 254 nm.Results The developed method had a good linear ( r>0.991 8 ) in the content range of 2-200 μg· mL-1 , with an aver-age recovery ranged from 88.5% to 123.4%.The precision was within 0.12%-3.42%.Conclusion PITC pre-column derivatization and HPLC method was developed and validated , which had good accuracy , suitable for the determination amino acid in serum and urine of patients with diabetes .

Objective:To observe the therapeutic effect of Shengsan Jiedu Huayu decoction in alleviating inflammatory liver injury in rats with acute-on-chronic liver failure (ACLF) and its effect on the activation intensity for the NLRP3 signaling pathway.Methods:63 SD rats were randomly divided into a blank group, a model group, and low-, medium-, and high-dose groups of Shengsan Jiedu Huayu decoction (7.29 g/kg/d, 14.58 g/kg/d, and 29.16 g/kg/d). The ACLF rat model was replicated using carbon tetrachloride combined with d-galactosamine and lipopolysaccharide. Different dose gradients of the Shengsan Jiedu Huayu decoction were used for a five-day intervention treatment, and then rat serum and tissue samples were collected. A biochemical analyzer was used to detect the serum levels of ALT, AST, and TBIL in rats. ELISA was used to detect serum IL-18 and IL-1β content. HE staining was used to observe histomorphological changes in liver tissue. Immunohistochemistry was used to detect GSDMD expression in liver tissue. Western blot and PCR were used to detect NLRP3, Caspase1, ASC, TLR4, IL-1β, IL-18 protein, and mRNA expression levels.The groups were compared using analysis of variance and the rank-sum test.Results:Compared with the blank group, the model group’s rat liver tissue was severely injured. Serum levels of ALT, AST, and TBIL, inflammatory factors IL-1β and IL-18, and the GSDMD protein expression level, NLRP3 expression level, TLR4, caspase 1, ASC, IL-1β, IL-18 protein, and mRNA ( P<0.01) were all significantly increased in the model than the blank group ( P<0.01). Additionally, compared with the model group, the low-, medium-, and high-dose groups of Shengsan Jiedu Huayu decoction had improved liver tissue injury in ACLF rats, while the serum levels of ALT, AST, TBIL, IL-1β, IL-18, liver tissue GSDMD protein, NLRP3, TLR4, caspase 1, and ASC expressions were all lower in the different dose gradients of the Shengsan Jiedu Huayu decoction than the model group, with the most evident reduction in the high-dose group ( P<0.01). Conclusion:Shengsan Jiedu Huayu decoction can weaken the activation intensity of the NLRP3 signaling pathway, alleviate liver tissue pathological injury, reduce inflammatory factor release, and alleviate inflammatory liver injury in ACLF rats.

OBJECTIVETo study the effects of calcium and calmodulin dependent kinase against hypoxic neuronal injury and its possible mechanisms.

METHODSEmbryonic cortical neurons of 17-day pregnant embryo Sprague-Dawley rats were cultured in vitro and the cultured neurons were randomly allocated into different groups that exposed to hypoxia or hypoxia +calcium channel antagonist. Nimodipine and MK-801 were used to block the L-voltage sensitive calcium channel and NMDA receptor respectively before hypoxia. The methyl thiazolyl tetrazolium (MTT) method was used to determine the cell viability. Fluo-4AM, an intracellular calcium indictor, was used to detect the changes of intracellular calcium after hypoxia. The expressions of CaMKII and CaMKIV were detected by Western blot.

RESULTSThe cell viability of the nimodipine or MK-801-treated groups was significantly higher than that of the untreated hypoxia group. The intracellular calcium level of the nimodipine-treated group decreased rapidly after hypoxia. Compared to nimodipine treatment, MK-801 treatment could inhibit hypoxia-induced calcium influx for a longer time. Nimodipine treatment decreased the CaMKII expression while MK-801 treatment decreased the CaMKIV expression.

CONCLUSIONSNimodipine and MK-801 protect neurons from hypoxic injury possibly by the inhibition of CaMKII and CaMKIV expressions respectively.

Animals ; Calcium ; analysis ; Calcium-Calmodulin-Dependent Protein Kinase Type 2 ; Calcium-Calmodulin-Dependent Protein Kinase Type 4 ; Calcium-Calmodulin-Dependent Protein Kinases ; antagonists & inhibitors ; physiology ; Cell Hypoxia ; Dizocilpine Maleate ; pharmacology ; Female ; Neurons ; pathology ; Neuroprotective Agents ; pharmacology ; Nimodipine ; pharmacology ; Rats ; Rats, Sprague-Dawley