Objective To investigate the expression of nuclear factor-κB in alveolar macrophages of paraquat-induced rats and the effect of diallyl sulfide on it.Methods Forty five male wistar rats were randomly divided into three groups,namely control group,model group,and DAS treatment group (n =15 in each).The model of paraquat poisoning was reproduced by single does of 70 mg/kg given by intra-gastric administration,while the equal volume of normal saline (NS) was given to the rats in control group instead.The dose of 100 mg/kg of DAS was given to rats by intra-peritoneal injection in DAS treatment group.The equal volume of NS was given to the rats by intra-peritoneal injection in model group and control group instead.The rats of model group and DSA treatment group were exposed to paraquat once a day for 14 days.Five rats in each group were sacrificed at 3,7,14 days.Alveolar maerophages were harvested by bronchalveolar lavage (BAL).The protein content of BAL fluid were examined.The exprossion of NF-κB was measured with immunocytochemistry technique.Results Alveolar macrophage cultures were carried out by using differential adherence of isolated and purified alveolar macrophages,and after 30 minutes culture,more adherent macrophages can be seen.Compared with model group,the protein content of BAL fluid at dfferent intervals in the control group were obviously lower,especially on the 3 rd day (261.6 ± 17.16) μg/mL vs.(673.4 ± 151.9) μg/mL;7 d (265.6 ± 18.37) μg/mL vs.(581.3 ± 134.58) μg/mL;14 d (253.8 ± 11.43) μg/mL vs.(589.07 ± 33.85) μg/mL,P ＜ 0.05.Comparisons of protein content in BLA fluid between PQ group and DAS treatment group were on the 3 rd day (673.4 ± 151.9) μg/mL vs.(342.9 ±39.03) μg/mL;on the 7 th day (581.3 ± 134.58) μg/mL vs.(383.7 ±7.37) μg/mL,P＜0.05;on the 14 th day (589.07±33.85) μg/mL vs.(282.9±15.59) μg/mL,P＜0.05.The immunocytochemistry analysis revealed minimal NF-κBp65 expression in the cell cytoplasm in the control group,while high NF-κBp65 expression was found in nuclear in the model group.Minimal NF-κBp65 expression was found in the cell cytoplasm in the DAS treatment group,and integral A value was significantly lower in the DAS treatment groups than that in the model group (P ＜ 0.05).Conclusions Treatment with an intra-peritoneal injection of DAS is capable of attenuating the extent of PQ-induced ALI in rats by lowering BLA fluid protein content,inhibiting the expression of NF-κB in alveolar maerophages.

Objective To investigate the relationship between the plasma inflammatory cytokines and carotid atherosclerosis in patients with ischemic cerebrovascular disease(ICVD).Methods The levels of plasma interleukin (IL)-6,matrix metalloproteinase (MMP)-8 and soluble CD40 ligand (sCD40L) in 64 patients with ICVD were detected by enzyme-linked immunosorbent assay (ELISA),the severity of bilateral carotid atherosclerosis was measured by colour ultrasound.The results were compared with those of non ICVD controls. The association of the levels of plasma IL-6,MMP-8,sCD40L and the severity of carotid atherosclerosis were analysized. Results The levels of plasma IL-6,MMP-8,and sCD40L in patients with ICVD were significantly higher than those in control group(all P

OBJECTIVETo observe the expression of peripheral blood CD4+ T lymphocyte apoptosis gene in rheumatoid arthritis (RA) patients with cold dampness type (CDT), and to explore its correlation with clinical indicators of RA.

METHODSSixteen RA patients with CDT (as the RA group) and 16 healthy subjects (as the normal control group) were recruited. CD4 T cell apoptosis rate was detected in the RA group and the normal control group using FCM. mRNA expressions of fas, fasL, caspase-3, caspase-8, bcl-2, and bax were detected using RT-PCR. Correlations between the expression of apoptosis gene and clinical activity indicators of RA (ESR, CRP, RF, CCP, integrals for Chinese medial symptoms, morning stiffness time, joint tenderness number, joint swelling number, DAS28-3) were analyzed.

RESULTSThe apoptosis rate of CD4+ T was significantly lower in the RA group than in the control group [(2. 6 +/- 0.9) % vs. (7.7 +/- 1.3) %, P < 0.01]. mRNA expression levels of fas, fasL, caspase-8, caspase-3, and bax mRNA of CD4+ T significantly decreased, but bcl-2 mRNA expression increased in the RA group (P < 0.01). The apoptosis rate of CD4+ T was negatively correlated with ESR (P < 0.05). The mRNA expression of caspase-8 was negatively correlated with joint swelling number (P < 0.05). The mRNA expression of bcl-2 was negatively correlated with integrals for Chinese medial-symptoms and joint function classification (P < 0.01, P < 0.05).

CONCLUSIONApoptosis obstacle exists in peripheral blood CD4 +T lymphocyte of RA patients, and is closely related to disease activity.

Apoptosis ; Arthritis, Rheumatoid ; diagnosis ; metabolism ; pathology ; CD4-Positive T-Lymphocytes ; Caspase 3 ; metabolism ; Caspase 8 ; metabolism ; Fas Ligand Protein ; Humans ; RNA, Messenger

Anti-Infective Agents ; administration & dosage ; therapeutic use ; Dental Plaque ; therapy ; Humans ; Periodontal Diseases ; drug therapy

Objective To investigate whether 5, 7-dihydrox-8-nitrochrysin (NOC) induces apoptosis in U937 monocytic leukae-mia cells is involved in the regulation of the activity of PKD 2.Methods U937 cell line cells were cultured in vitro .Apoptosis rate was analyzed by flow cytometry (FCM) using propidium iodide (PI) staining.DNA ladder bands were observed by DNA agarose gel electro-phoresis.The phosphorylated protein expression of PKD 2 was analyzed using Western blot .Results NOC (2.5, 5.0, and 10.0μmol/L) increased apoptosis rate in U937 cells in a concentration-dependent manner ( P <0.05).After treatment with NOC (5.0 and 10.0μmol/L) for 24 h, U937 cells presented typical DNA ladder bands .At the same time, not only did NOC effectively down-regulate the ex-pression of PDK2 phosphorylated protein , but also increased apoptosis rate in U 937 cells in the presence of G?6976, a specific inhibitor of PKD2.Conclusions The effect that NOC induces apoptosis in U 937 cells is related to the inhibition of the activity of PKD 2.

Objective To study the inlfuence factors on detection of activity of four coagulation factors in prothrombin complex concentrates (PCC) by several factors. Methods Using Pharmacopoeia of the People’s Republic of China (2010) as reference, the activity of four coagulation factors in PCC were investigated by choosing different pre-treatments, different diluents, different salt concentration, different standard human plasma and different company reagents. Results The activity of FII, FVII, FX were decreased and FIX was increased in the condition of adding protamine sulfate, and there were no differences of four coagulation factors whether warm bath in 37 for 15 min or not. However, the differences of four coagulation factors were significant by using deficient plasma, saline, distilled water and commercial dilution buffer(P<0.05). The activity of coagulation factor II, X in 1 mol/L salt concentration of PCC were significantly lower than in 0.25 mol/L(P<0.05), while coagulation factor VII, IX were not. The activity of FII, FVII, FIX, and FX were different by using different standard human plasma to make standard curve. The activity of four coagulation factors existed significant difference(P=0.00) by using SIEMENS company reagents and domestic reagents. Conclusion Choosing different pre-treatments, different dilution buffers, salt concentration, standard human plasma and commercial kits will inlfuence the detection result of coagulation factors.

BACKGROUND: Serum uric acid level is associated with some chronic diseases and prognosis of severe infection. This study aimed to investigate the relationship between serum uric acid (SUA) and prognosis of infection in critically ill patients. METHODS: The data from 471 patients with infection admitted from January 2003 to April 2010 were analyzed retrospectively at Huashan Hospital Affiliated to Fudan University, Shanghai, China. The data of SUA, serum creatinine, blood urea nitrogen (BUN) and other relevant examinations within 24 hours after admission were recorded and the levels of SUA in those patients were described, then Student's t test was used to evaluate the relationship between SUA and pre-existing disorders. Different levels of SUA were graded for further analysis. The Chi-square test was used to examine the difference in the prognosis of infection. RESULTS: The mean initial level of SUA within 24 hours after admission was 0.232±0.131 mmol/L and the median was 0.199 mmol/L. Remarkable variations in the initial levels of SUA were observed in patients with pre-existing hypertension (t=–3.084,P=0.002), diabetes mellitus (t=–2.487, P=0.013), cerebral infarction (t=–3.061,P=0.002), renal insufficiency (t=–4.547,P<0.001), central nervous system infection (t=5.096,P<0.001) and trauma (t=2.875,P=0.004). SUA was linearly correlated with serum creatinine and BUN (F=159.470 and 165.059, respectively,P<0.001). No statistical correlation was found between the initial levels of SUA and prognosis of infection (χ2=60.892,P=0.100). CONCLUSION: The current study found no direct correlation between the initial levels of SUA after admission and prognosis of infection in critically ill patients.

Object To distinguish the genuine Dioscorea polystachya Turcz. from other species of Dioscorea L. such as D. persimilis Prain et Burkill, D. japonica Thunb., and D. alata L., conventionally used in some localities by sequencing their nuclear ribosomal RNA subunit (18S rRNA), to provide mole cular evidences for the quality control of drugs of Dioscorea L. Methods By direct PCR sequencing to detect the homology of 18S rRNA sequence of different species of Dioscorea L.. Results The 18S rRNA gene sequence of genuine D. polystachya, D. persimilis and D. japonica showed identical length of 1810 bp, whereas that of D. alata was 1807 bp. Multiple sequence alignment of 18S rRNA gene showed that the homology was identical between D.persimilis and the genuine D. polystachya; but a difference of 99.89% with D. japonica and 97.51% with D. alata. Conclusion DNA sequencing can provide an accurate and reliable mean for the identification of the origin and genuineness of Dioscorea L..

Objective To investigate the effect on cell cycle of hepatitis B virus x protein and ap- proach the molecular mechanism of x protein's carcinogenesis. Methods Gene transfection mediated by the lipofectamine was used to introduce the eukaryotic expression vector of HBV x gene into human hepato- cellular carcinoma cell line HepG2(HepG2X cell).The selective medium containing G418 was used to select the cell clones which the X protein was expressed constantly.Flow eytometry was used to detect the cycle of the cells;raf-1 protein was detected by Western blot.Results X protein could be expressed on 21?10~3 loca- tion in the transfected cells,while there were no protein expressed in the control cells.The proliferation of X cells increased significantly according to MTS method(P