OBJECTIVETo evaluate therapeutic effect of Sishen Wan on experimental colitis, and explore its mechanism by expression of Bax/Bcl-2 mRNA, Fas/FasL in colonic tissue.

METHODExperimental colitis was induced by rectal administration of trinitrobenzene sulfonic acid (TNBS) dissolved in ethanol. The model animals were divided into four groups: the induced colitis but untreated group, the induced colitis groups treated with the high, middle, low dose of Sishen Wan, and the induced colitis group treated with salicylazosulfapyridine (SASP). After 10 day administration, the body weight, colonic wet weight, colonic weight index, colonic damage score and pathological change were evaluated, and the level of Fas and FasL by flow cytometry, Bax mRNA and Bcl-2 mRNA by reverse transcription polymerase chain reaction (RT-PCT).

RESULTCompared with the model group, the colonic wet weight and colonic weight index were remarkably decreased in the middle dose of Sishen Wan group (P < 0.05). The colonic injury scores were significantly reduced after rats were treated with the three doses of Sishen Wan (P < 0.05). Representative restored features were observed including fewer inflammatory cellular infiltration and follicular hyperplasia, superficial and little ulcer with fibroplasia in colonic mucosa from the treated groups. The expression of Fas in the colonic mucosa was obviously down-regulated (P < 0.05) and the ratio of Bcl-2 mRNA/Bax mRNA was significantly up-regulated (P < 0.05) in the groups treated with the three doses of Sishen Wan.

CONCLUSIONSishen Wan might postpone colonic epithelium apoptosis or improve inflammatory cell apoptosis by regulating the expression of Fas/ FasL and Bax/Bcl-2 mRNA in colonic tissue, which is possible potential path to effectively treat experimental colitis by enema.

Animals ; Colitis ; drug therapy ; metabolism ; Colon ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Fas Ligand Protein ; genetics ; Female ; Male ; Proto-Oncogene Proteins c-bcl-2 ; genetics ; RNA, Messenger ; analysis ; Rats ; Rats, Sprague-Dawley ; bcl-2-Associated X Protein ; genetics ; fas Receptor ; genetics

OBJECTIVETo observe effect of oral scorpio and scolopendra powder on T-cell subsets in peripheral blood and intestine from rats with collagen induced arthritis (CIA).

METHOD60 rats were randomly divided into 6 groups: normal control group, model control group, low-dose scorpio and scolopendra group, middle-dose scorpio and scolopendra group, high-dose scorpio and scolopendra group, and type II collagen group. Rat's rheumatoid arthritis was induced by collagen II (C II). Level of T-cell subsets from peripheral blood and intestine was measured by flow cytometry.

RESULTCD4+ T cellular level was obviously increased (P < 0.05 or P < 0.01) or kept increased tendency in peripheral blood and intestine from the model group compared with that of the normal group, while the ratio of CD4+/CD8+ in intestine was obviously descent but the contrary in peripheral blood (P < 0.05 or P < 0.01). CD4+, CD8+ T cellular level in intestine were obviously descent and the ratio of CD4+ /CD8+ increased in all treated groups when compared with in the model group (P < 0.05 or P < 0.01). However, CD4+ T cellular level and the ratio of CD4+/CD8+ in peripheral blood were remarkablely decreased.

CONCLUSIONThe mechanism that scorpio and scolopendra could treat rat's rheumatoid arthritis may be regulating balance of T-lymphocyte subsets in peripheral blood and intestine.

Animals ; Anti-Inflammatory Agents ; pharmacology ; Arthritis, Experimental ; immunology ; Arthritis, Rheumatoid ; immunology ; CD4 Lymphocyte Count ; CD4-Positive T-Lymphocytes ; CD8-Positive T-Lymphocytes ; Female ; Intestinal Mucosa ; immunology ; Medicine, Chinese Traditional ; Rats ; Rats, Wistar ; Scorpions ; chemistry ; T-Lymphocyte Subsets ; drug effects ; immunology

Objective: To study the biological effects of the HPV16Z-Hsp65-E6/E7 fusion protein vaccine on the tumor associated with HPV16 infection. Methods: We tested the cellular immune responsive intensity to the vaccine by the lymphocyte proliferation and CTL response, and studied the therapeutic effect of the vaccine on mouse TC-1 cell transplanted cancer in vivo and the influence on mouse lifetime. Results: The spleen lymphocytes from the C57BL/6 mouse immunized by the Hsp65-E6/E7 vaccine could proliferate obviously in the presence of the protein and TC-1 tumor cell could be lysed specifically by immune activated lymphocytes in vitro. This animal therapeutic experiment in vivo showed that the vaccine suppressed the growth of TC-1 cell transplanted tumor remarkably. Conclusion: The recombined vaccine can induce specific cellular immune response in vitro and suppress HPV16 positive TC-1 tumor cell growth obviously in vivo.

Objective To investigate the heterogeneity of chemosensitivity in colorectal cancer using an ATP-tumor chemosensitivity assay (ATP-TCA) and the feasibility of individual chemotherapy.Methods An ATP-TCA were used to determine the effect of 16 single or combined cytotoxic drugs in surgical specimens from 50 patients with colorectal cancer.Results There were considerable differences in chemosensitivity between individuals.The most active single drugs in the assay was identified as Navelbine, Hydroxycamptothecin, 5-Fluorouracil and Paclitaxel; 34.1%, 31.6%, 27.6% and 24.3% of specimens showed sensitivity to them, respectively.5-Fluorouracil+Mitomycin+Aytarabine was found to be the most effective combination, for 100% (11/11)specimens were sensitive to this regimen.5-Fluorouracil+Cisplatin+Adriamycin and Gemcitabine+Cisplatin were moderately active regimens.Conclusions There was the heterogeneity of the in vitro chemosensitivity in colorectal cancer.The use of the ATP-TCA provides a method of selecting appropriate anti-cancer drugs in colorectal cancer.

Objective: To develop a therapeutic adjuvantfree protein vaccine against HPV16 which is closely related to cervical cancer of China. Method: First the E6/E7 gene by PCR technology from HPV16z virus strain was isolated in the highrisk cervical cancer area of Shanxi province of China in1990s, and again got the gene segment of Hsp65 from BCG by the same method, mutated the transforming codes in sequences of HPV 16 E6/E7 genes and thus constructed the expression vector pET28aHsp65E6/E7, expressed the Hsp65E6/E7 fusion protein in E.coli BL21(DE3) strain and researched optimal protein purification procedures. Results: The expression vector pET28aHsp65E6/E7 was constructed successfully and E6/E7 gene was mutated correctly. Hsp65E6/E7 fusion protein was renatured and purified on the affinity chromatography column simultaneously. The protein purity achieved 95% after the anionic exchange chromatograph purification. conclusions: This research laid a foundation for further functional study of the therapeutic adjuvantfree protein vaccine——Hsp65E6/E7.

Objective To observe the efficacy and safety of pigolitazone/metformin fixed-dose combination therapy replacing metformin alone or combined with other anti-diabetes drugs in type 2 diabetes with poor glycemic control.Methods 80 cases were recruited,with an average age of (54.79±13.99)years,diabetes history of (9.76±6.59) and baseline HbA1c (9.06±1.34)％.All participants received pigolitazone/metformin instead of metformin without other treatment changes.Glycemic control (level of fast blood glucose,HbA1c) was evaluated at 12 weeks,as well as lipid profiles,liver and renal function,adverse events and body weight.Results 8 cases were lost to visit,4 cases were withdrawn for edema,only 68 subjects finished the study.Compared to the baseline,after 12-week treatment,FPG decreased for (2.06+0.16) mmol/L,HbA1c decreased for (0.84+0.23)％,both of the differences were statistically significant (P＜0.001,P＜0.001).Body weight increased (0.34+1.13)kg,with no difference compared to the baseline.The lipid profile presented elevated high density lipoprotein cholesterol (P=0.012)and decreased total cholesterol,low density lipoprotein cholesterol,triglyceride,while the latter three items showed no differences (P＞0.05,P＞0.05,P＞0.05).Indexes reflecting liver and renal function,such as ALT,AST,TBIL,DBIL,Urea,UA,Cr showed no differences compared with the baseline.Adverse events analysis showed at the end of the study,no severe hypoglycemia and serious cardiovascular events occurred,6 cases suffered edema,among whom 4 patients exited the study for severe lower limb edema.No extra gastrointestinal symptom happened.Conclusion Pigolitazone/metformin fixed-dose combination exhibits an excellent efficacy and safety for T2DM,with satisfying tolerability and compliance,which is a selection for those patients with poor glycemic control.

Objective To compare the therapeutic effect of postprandial and preprandial injection of glulisine.Methods Sixty hospitalized patients with T2DM receiving one dose of glargine and three doses of glulisine were recruited.They were randomly divided into two groups:group A and group B when the glycemic state and insulin dosages had been stable for more than seven days.Two-stage cross design:stage 1:group A (n=30):glulisine was injected before meal;Group B (n=30):glulisine was injected after meal.Blood glucose was monitored for three days.Stage 2:glulisine was injected after meal in group A while before meal in group B without dosages adjustment,and blood glucose was monitored continuously for three days.Then standard deviation of blood sugar (SDBG),blood glucose fluctuation after meal (PPGE),maximum blood glucose fluctuation range (LAGE) during 24 hour and satisfaction values of insulin treatment (SVIT) were compared.Results There was no significant differences between group A and group B in terms of age((50.70±13.29)years vs (55.63±13.05) years,P=0.152),diabetes course((36.23±29.20)months vs (43.63±32.19) months,P=0.355),HbA1c ((10.05％± 1.46％)vs (9.81％±2.08％,P=0.612),daily insulin dose((35.67±8.64)U vs (34.83±8.24) U,P=0.704),SDBG ((2.63±0.58 vs (2.84±0.64)) before operation.There was no significant differences of SDBG(F=0.432,P=0.73),PPGE (F=1.216,P=0.31),LAGE (F=0.431,P=0.73) or SVIT (F=0.685,P=0.56) between glulisine injected before and after meal.Conclusion Postprandial glulisine administration can provide the same effect in lowering glucose,satisfaction values and reducing glucose fluctuation as preprandial injection.

Objective:To explore the value of a radiomics nomogram based on T 1WI for prediction of the relapse of osteosarcoma after surgery within 1 year from multicenter data. Methods:The imaging and clinical data of 107 patients with pathologica1ly confirmed osteosarcoma who received neoadjuvant chemotherapy before surgery from 6 hospitals from January 2009 to October 2017 were retrospectively analyzed. A training cohort consisted of 75 patients from firstly enrolled 4 hospitals and an independent validation cohort of 32 patients from other 2 hospitals. Pretreatment T 1WI was used to extract radiomics features. Least absolute shrinkage and selection operator (LASSO) regression was applied to reduce the dimension and then the radiomics signature was constructed to predict the relapse of osteosarcoma after surgery within 1 year in training cohort. Independent clinical risk factors were screened using one-way logistic regression, and then a radiomics nomogram incorporated the radiomics signature and MRI characteristics was developed by multivariate logistic regression. The predictive nomogram was evaluated using receiver operating characteristic (ROC) curve in the training cohort, and validated in the independent validation cohort. The calibration curve was used to evaluate the agreement between prediction and actual observation and the decision curve was used to demonstrate the clinical usefulness. Results:Based on T 1WI from multicenter institutions, the radiomics signature was built using 2 valuable selected features that were significantly associated with relapse within 1 year. Two selected features included 1 gray-level co-occurrence matrices (GLCM) feature (L_G_1.0_GLCM_homogeneity1, LASSO coefficient 3.122) and 1 gray-level run length matrix (GLRLM) feature (GLRLM_RP, LASSO coefficient -2.474). The prediction nomogram including radiomics signature and MRI characteristics (joint invasion and perivascular involvement) showed good discrimination with the area under the ROC curve of 0.884 and 0.821 in the training and validation cohorts, respectively. The calibration curve showed that the nomogram achieved good agreement between prediction and actual observation. Decision curve analysis demonstrated that the radiomics nomogram was clinically useful when the threshold probability was greater than 21%. Conclusion:The radiomics nomogram based on T 1WI can be used as a non-invasive quantitative tool to predict relapse of osteosarcoma within 1 year before treatment, which provides support for clinical decision-making in osteosarcoma.