Objective To investigate the effect of rapamycin(RPM)on hepatic interleukin-10(IL-10)gene and acute liver injury in rats with postburn Staphylococcus aureus sepsis.Methods Fifty male Wistar rats were randomly divided into four groups:normal control group,scald control group,postburn sepsis group,and RPM treatment group.Tissue samples from liver and plasma were collected to determine IL-10 mRNA and protein expressions,and liver function parameters were also measured.Results Compared to postburn Staphylococcus aureus sepsis group,in RPM treatment group hepatic IL-10 mRNA expression and plasma IL-10 were significantly increased at 0.5 hour after RPM treatment(P

To observe the effect and its potential mechanism of monoclonal antibody to tumor necrosis factor-alpha(TNF? MoAb)on systemic hemodynamics and survival rate after intestinal ischemia/reperfusion (Ⅱ/R). Method: SD rats were subjected to 75 rain of superior mesenteric artery occlusion followed by 6 hours of reperfusion. The animals were treated intravenously with either TNF? MoAb(20mg/kg)or the control protein(albumin, 20mg/kg) 30 min prior to the onset of ischemia. Result: Pretreatment with TNF? MoAb significantly attenuated the decrease in blood pressure and cardiac index compared to controls throughout the 6-hour period of observation(P

To evaluate the effect of delayed treatment with monoclonal antibody to tumor necrosis factor-alpha (TNF-? MAb) on systemic hemodynamics and multiple organ dysfunction following prolonged hemorrhagic shock and resuscitation. Method: Adult male Sprague-Dawley rats were subjected to prolonged hemorrhagic shock (MAP of 4.00-4.66 kPa for 180 rain)followed by resuscitation over 50 min. The animals were treated intravenously with either TNF-? MAb (20.0 mg/kg) or the control protein(albumin,20.0 mg/kg)15 rain after the end of resuscitation(65 min after shock). Result: Compared to the albumin controls, delayed treatment with TNF-? MAh significantly reduced the total peripheral resistance index (P

Objective To investigate the effect of nuclear factor-kappa B (NF-?B) signal transcription pathway inhibitor-pyrrolidine dithiocarbamate (PDTC) on mRNA expression of high mobility group box-1 protein (HMGB1) in organs of rats with endotoxic shock and its potential mechanism. Methods Forty-seven male Wistar rats were randomly divided into normal control group (n=8), endotoxic shock group (n=24), and PDTC treatment group (n=15). At serial time points, the animals in each group were sacrificed, and tissue samples from the liver, lungs and kidneys were harvested to determine HMGB1 mRNA expression and pulmonary MPO activity. Also, blood samples were collected to determine the major organ functional indices. Results Compared to normal controls, HMGB1 mRNA levels were significantly increased in samples from the liver, lungs and kidneys at 2-6h after endotoxin challenge (P

Objective To investigate the effect of lipopolysaccharide (LPS) on stimulating high mobility group box-1 protein (HMGB1) mRNA expression in peritoneal macrophages in rats and its potential signaling mechanism. Methods Abdominal macrophages obtained from male Wistar rats were seeded on 24-well (1?10 6 cells/well) tissue culture plates. The cells were incubated for 3 days before they were stimulated with LPS, fludarabine (specific inhibitor of signal transducer and activator of transcription 1, STAT1), or rapamycin (specific inhibitor of signal transducer and activator of transcription 3, STAT3). After being stimulated, macrophages were denatured directly in tissue culture plates to determine HMGB1 mRNA expression. The time-dependent and dose-dependent responses between LPS stimulation and HMGB1 mRNA expression were analyzed, and the effect of treatment with fludarabine and rapamycin on HMGB1 mRNA expression was also observed. Results After being stimulated with 75-100ng/ml LPS, the HMGB1 mRNA expressions in macrophages were up-regulated markedly, peaking at 24-36 hours, and remained elevated up to 48 hours. It was found that the HMGB1 mRNA expression was significantly inhibited by treatment with either fludarabine (100?mol/L) or rapamycin (25ng/ml). Conclusions These data suggest that LPS stimulation can result in up-regulation of HMGB1 mRNA expression in peritoneal macrophages in rats. Janus kinase/STAT pathway may be involved in modulating HMGB1 mRNA expression in LPS-activated macrophages.

The lineage-Actinobacteria class nov.comprises organisms with a DNA base composition which generally is above 50% G+C (with a few exceptions).We set up a method that has been used in isolating Actinobacteria and Actinomycetes. We added 25?g/mL Nalidixic acid and 25?g/mL Aztreonam into isolation media to inhibit the other bacteria and 20?g/mL Benlete to inhibit fungi.We used fluorescent in situ hybridization to identi 1fy Actinobacteria. Using the four probes,PA-1,PA-2,PHGC and PNHGC,we made the identification on the 31 strains of the 56 gram-positive bacteria randomly selected and got 22 positive results,6 negative results and 3 ambiguous results.It was showed that the results of G+C content determination and FISH method were identical.Among 31 strain,there were 24 strains of Actinobacteria,the rate was 77.4%.This proved the isolation and FISH identification methods were effective and reliable.

Objective To establish the quality control standard for Shuanggen Qingnao Granules.Methods TLC was used to identify Yiyiren SEMENCOICIS,Yujin RADIXCURCUMAE,Banlangen RADIX ISATIDIS. HPLC was used to determine the content of Puerarin in Shuanggen Qingnao Granules.Results The TLC spots developed were fairly clear,and the bland test showed no interference.Puerarin showed a good linear relationship in the range of 5.184～40.50?g/mL,the average recovery of Puerarin was 99.93%,and RSD=1.81%.Conclusion The method is accurate and quick,and can be used for the quality control of Shuanggen Qingnao Granules.

Objective To investigate the correlation between the Periostin and Epithelial-Mesenchymal Transition(EMT) markers(E-cadheirn and N-cadheirn) in colorectal cancer(CRC),and analyze the relationship between the expression of Periostin and clinicopathological parameters.Methods The expression of Periostin,Ecadheirn and N-cadheirn in 106 cases with primary CRC tumors and corresponding normal tissues were detected by Immunohistoehemistry and the results were analyzed.Results The expression of Periostin,E-cadheirn and N-cad-heirn in tumors were significantly high expression than those in corresponding normal tissues(62/106 vs 21/106,x2 =34.027,P ＜ 0.05 ;43/106 vs 89/106,x2 =42.480,P ＜ 0.05 ; 66/106 vs 19/106,x2 =43.382,P ＜ 0.05).The expression of Periostin in tumors was associated significantly with tumor differentiation,tumor invasion,lymph node metastasis and distance metastasis (x2 =7.752,P =0.007; x2 =5.008,P =0.031 ; x2 =10.227,P =0.002;x2 =8.001,P =0.006).It was negatively correlated with E-cadherin expression (r =-0.435,P ＜ 0.001),but positively correlated with N-cadherin expression (r =0.213,P =0.028).Conclusion Periostin may promote the occurrence and development of CRC by participating in EMT program.

Adult ; Aged ; Asteraceae ; Drugs, Chinese Herbal ; adverse effects ; Female ; Hepatic Veno-Occlusive Disease ; chemically induced ; Humans ; Male ; Middle Aged ; Retrospective Studies

Objective To observe the expression of tumor necrosis factor-α induced protein 8 like-2 (TIPE2) in CD4 + CD25 + regulatory T cells ( CD4 + CD25 + Tregs) and analyze its potential effect on immunosuppressive activity of CD4 + CD25 + Tregs. Methods CD4 + CD25 + Tregs were purified from spleen of the BALB/c mice by using magnetic cell sorting system.The expressions of TIPE2 mRNA and protein in CD4 + CD25 + Tregs were detected by using RT-PCR and Western blot.CD4 + CD25 +Tregs were further infected with the recombinant lentiviruses that carried small interference RNA(siRNA)to knock down the TIPE2 expression.Based on the expressions of cell surface molecules including cytotoxic T-lymphocyte-associated antigen (CTLA)-4 and forkhead/winged helix transcription factor p3 (Foxp3) detected by flow cytometry and the levels of cytokines including interleukin (IL)-10 and transforming growth factor (TGF)-3 examined by ELISA in CD4 + CD25 + Tregs,the functional role of TIPE2 in controlling suppressive activity of CD4 + CD25 + Tregs was analyzed.In the meantime,the proliferation activities of T effector cells were assayed by MTT test. Results A 147 bp TTPE2 gene band and a clear TIPE2 band with a molecular mass of approximately 21 000 in CD4 + CD25 + Tregs were detected by using Westem blot.Cell surface molecule as well as cytokine expressions were significantly down-regulated when the CD4 + CD25 + Treg stimulated and activated by anti-CD3/CD28 was cultured for 24 hours after the siRNA silenced CD4 + CD25 + Treg cells ( P ＜ 0.01 ).Meanwhile,the suppression role of CD4 +CD25 + Tregs on the proliferation activity of T effector cells was weakened obviously ( P ＜ 0.05 ). Conclusions As an important immunoregulatory molecule,TIPE2 not only expresses in the CD4 + CD25 +Tregs,but also affects the immunosuppressive function of CD4 + CD25 + Tregs.