Objective: To study the effect of phytic acid (PHA) on 1,2-dimethylhydrazine (DMH) induced colorectal carcinogenesis in rats. Methods: Thirty four weeks old healthy male Wistar rats were randomly divided into two groups,control group given basal diet and PHA group given basal diet plus 2% sodium phytate solution. Both groups were injected DMH weekly for 20 w and then killed . Body weight, mortality, tumour incidence and average size, and natural killer(NK)cell activity were measured. Result: The mortality was lower in PHA group (P

Objective: To investigate the effects and mechanisms of inositol hexaphosphate(IP6)on cell cycle of HT-29 human colon carcinoma cell line. Method: HT-29 cells were exposed to various concentrations of IP6 for certain time. The effect of IP6 on cells proliferation was evaluated by MTT assay. Flow cytometric analysis was performed for cell cycle progression. Immunocytochemical stain was used to detect the expression of p53 and p21 protein. Results: A significant dose-dependent as well as time-dependent growth inhibition was observed in IP6-treated HT-29 cells, and associated with an increase in G1 arrest; Different concentrations of IP6 decreased the abnormal expressions of p53 gene and strongly increased the expression of p21. Conclusions: IP6 had an inhibitory effect on proliferation of HT-29 cells .and its mechanisms might be related to many factors, such as inhibiting the abnormal expression of p53 gene, inducing the expression of cell cycle inhibitor p21 which block the cell cycle.

Objective: To evaluate the effect of dietary fiber and phytic acid inositol hexaphosphate (IP6 or Insp6) on 1,2-dimethylhydrazine(DMH) induced colorectal carcinogenesis in Wistar rats. Methods: 86 four-week-old male Wistar rats were divided randomly into 6 groups and fed with either basal fiber free diet or the basal diet supplemented with 10% pectin, 10% cellulose, 2% Na-InsP6 supplemented water, 10% pectin in combination with 2% Na-InsP6 supplemented water, 10% cellulose in combination with 2% Na-InsP6 supplemented water. After four weeks, the rats were given a weekly injection of DMH (20mg/kg bw) for 20 weeks. The rats were killed after 26 weeks. The number of total macroscopically visible neoplasms was counted and the volume of individual neoplasmas was calculated. Proliferation cell nuclear antigen (PCNA) was analyzed. Results: There was no significant difference between the treatment group and the control group in the incidence of tumors. InsP6 significantly reduced the number of tumors in rats and tumor volume. Pectin and Pectin+InsP6 significantly increased the number of tumors in rats. The expression of proliferation marker PCNA was significantly down-regulated by InsP6 and significantly increased by pectin. Conclusion: A treatment regimen of 2% Na-InsP6 in drinking water was effective in significantly reducing the risk of colorectal cancer in Wistar rats. Diet supplemented with pectin could increase the risk of colorectal cancer.

BACKGROUND:Bone marrow mesenchymal stem cells(BMSCs)differentiating into neural cells is an effective way of cell therapy of nervous system disease.However,the methods used nowadays still need to be improved.OBJECTIVE:To induce the differentiation of rhesus monkey BMSCs into neuron-like cells by using sonic hedgehog factor.METHODS:Rhesus monkey BMSCs differentiating into neuron-like cells was induced by typical retinoic acid and sonic hedgehog factor.Rhesus monkey BMSCs were isolated and cultured by density gradient centrifugation method.Cell growth was observed under an inverted phase contrast microscope and cell growth curve was determined using MTT assay.Flow cytometry was performed to characterize the phenotype of BMSCs,and immunohistochemistry was utilized to assess differentiated cells.Ultra-structure of the differentiated cells was observed by transmission and scanning electron microscopes.RESULTS AND CONCLUSION:Rhesus monkey BMSCs cultured in vitro were identified by flow cytometry,with high homogenicity.Following sonic hedgehog factor disposal for 7 days,differentiated cells were mainly positive for neurone specific enolase,neurofilament protein,Tau and glial fibrillary acidic protein(GFAP).Image statistical analysis found that in sonic hedgehog factor scheme,neural stem cells marker Neetin positive rate was significantly higher compared with the rstinoic acid scheme(P＜0.01).GFAP-positive rate was greater in the retinoic acid scheme than in the sonic hedgehog factor scheme(P＜0.05).Results indicated that sonic hedgehog factor scheme is an effective pathway of rhesus monkey BMSC differentiation into neuron-like cells.

BACKGROUND: Present studies have demonstrated that during neural development, differentiation of neural stem cells (NSCs) was affected by various regulatory factors from surrounding microenvironment. Sonic hedgehog (Shh) is a key induction signal during neural fetal development, and can be an effective inductor to regulate differentiation of neural cells. OBJECTIVE: To investigate the signal transduction pathway of SHH for differentiation of rhesus monkey bone marrow mesenchymal stem cells (BMSCs) into of neuron-like cells by sonic hedgehog factor. METHODS: Rhesus monkey BMSCs were isolated and cultured by conventional density gradient centrifugation. BMSCs in the induction group were treated with L-DMEM containing FGF2, B27 and fetal bovine serum for preinduction of 24 hours, and then with DMEM supplemented with 0.5 μmol/L retinoic acid or 400 μmol/L SHH for 8 days. Non-induced cells served as control group. Following labeling with neuron enolase, positive cells were screened by flow cytometry. RT-PCR and Western-blot were used to detect SHH- and retinoic acid-induced cell membrane receptor and intracellular signal protein changes. RESULTS AND CONCLUSION: SHH specific membrane receptor Ptc, retinoic acid specific receptor RARα, signal protein molecule ptch1 and Smad expressed in normal cells. Ptc expression upregulated in SHH-induced cells. High expression lasted for a long time with induction time, which was significantly stronger compared with the retinoic acid and control groups (P < 0.01). Intracellular ptch1 protein molecule expression showed similar tendency as this, but could not induce upregulation of RARα expression. During induction, retinoic acid-stimulated cells did not activate Ptc pathway. Four days following induction, RARα expression upregulated and lasted till 6 days, but there were no significant differences. No significant change in ptch1 expression was determined. SHH- and retinoic acid-induced cell Smad molecule expression upregulated, but no significant difference was determined. Results verified that SHH-induced scheme participated in cell induction and differentiation by persistently activating its specific receptors. However, there was no significant receptor pathway crossing between retinoic acid-induced and SHH-induced schemes.

An evaluation system including experiment preparation,experiment process,com-prehensive design of experiment,experiment skills and written exam was established in order to adapt to the experimental teaching reform in our school. Experimental preparation assessment was to evaluate‘preview and self-evaluation report’prepared and submitted by students. Experimental process assess-ment was to evaluate students' classroom performance and experiment reports. Assessment of compre-hensive design experiments was to evaluate the overall participation of students. Skill assessment was consisted of oral test and experiment operation test. Final written examination,mainly consisting of subjective questions,emphasized on student's flexible use of knowledge and ability to solve practical problems. The evaluation system of promoting student's learning and teacher's teaching through the examination not only fully arouse student's attention on experiment,but also make teachers more ob-jectively and really understand students' learning situation and the teaching effectiveness.

OBJECTIVE:To standardize the clinical use of adjuvant therapy drugs. METHODS:To establish the clinical use management mode of adjuvant therapy drugs in our hospital and summarize the usage amount and irrational drug use of adjuvant therapy drugs before and after implementing the management,and the management effect was assessed. RESULTS:The manage-ment mode of adjuvant treatment drugs in our hospital included ensuring the directory for adjuvant therapy drugs,establishing the management system and ensuring its basic principles,then conducting drug use review with the ranking of drug dosages and mak-ing full use of evidence-based medical evidence,as well as developing principle of classification disposal for the irrational drug use (off-indications,overdose,ultra treatment) of adjuvant therapy drugs. After more than one year of management practice,the use of adjuvant therapy drugs in our hospital is increasingly standardized. The sales amount of adjuvant therapy drugs to all drugs was reduced from 20.90％ before management(Aug. 2014-Jul. 2015)to 15.99％(reduced by 5％)after management(Aug. 2015-Jul. 2016). Compared with Aug. 2015,the sales amount of 48 varieties in the 93 adjuvant therapy drugs was dropped more than 15％in Aug. 2016,and irrational drug use decreased obviously. CONCLUSIONS:The established management mode of adjuvant thera-py drugs in our hospital has effectively improved clinical rational drug use and reduced medical costs.

6cm,the recrudescenc rates when factors were male,mild or moderate malignant,stage of Dukes C,distance from tumor to anus 4～6cm were significantly higher(?2=4.53,P

BACKGROUND: Incidence rate of carcinoma of large intestine increases with the development of national living standard. Many epidemiologic surveys have showed that dietary factors have closely correlation with high risk of carcinoma of large intestine.OBJECTIVE: To observe the effect of dietary fiber and inositol hexaphosphate on the incidence of carcinoma of large intestine by establishing rat models with carcinoma of large intestine.DESIGN: Randomized block design.SETTING: Institute of Nutriology, Medical College, Qingdao University.MATERIALS: The experiment was performed at the Institute of Nutriology, Medical College, Qingdao University from March to December 2004.Totally 86 male Wistar rats of 4 weeks were randomly assigned based on body mass into cellulose group with 14 rats, pectin group with 14 rats, inositol hexaphosphate group with 15 rats, cellulose plus inositol hexaphosphate group with 14 rats, pectin plus inositol hexaphosphate group with 14rats and control group with 15 rats.METHODS: Basic feed without dietary fiber was given in the control group. 10% pectin was added in the pectin group. 10% cellulose was added in the cellulose group. 2% sodium phytate was added in the water of inositol hexaphosphate group. 10% pectin and 2% sodium phytate were added in the water of pectin plus inositol hexaphosphate group. 10% cellulose and 2% sodium phytate drinking water were added in the cellulose plus inositol hexaphosphate group. A total of 86 rats received 1,2-dimethylhydrazine (DMH) by subcutaneous injection to induce carcinoma of large intestine at week 4 after breeding. Incidence rate of large bowel neoplasm,number and volume of tumor were observed. Proliferating activity of large intestinal mucosa cells was determined (the ratio of number of positive cells of proliferating cell nuclear antigen to total count of nucleus).MAIN OUTCOME MEASURES: ①Incidence rate of large bowel neoplasm in each group, changes of mean number and mean volume of tumors of each rat, and ②proliferating activity of large intestinal mucosa cells of rats.RESULTS: ①Most of the rats in each group died during the 20-week trial. Oue rat died in the pectin group, the pectin plus inositol hexaphosphate group and the control group, respectively after 20 weeks. ②There was no significant difference in the incidence rate of large bowel neoplasm of rats compared with the control group (P ＞ 0.05), but the mean number and mean volume of tumors of each rat in the inositol hexaphosphate group were significantly lower than those in the control group [quantity: (1.1±0.2),(4.1±1.2)rats,P ＜ 0.01 ;volume: (176.1±65.5), (1 046.7±469.0)mm3,P ＜ 0.05], and the mean number of tumors of each rat in the pectin plus inositol hexaphosphate group was obviously higher than that in the control group [(7.5±1.9), (7.2±1.0) rats,P ＜ 0.05]. ③The proliferating activity of large intestinal mucosa cells in the inositol hexaphosphate group significantly decreased as compared with the control group. The proliferating activity of large intestinal mucosa cells in the pectin group increased as compared with the control group [(41.8±4.7)%, (83.6±2.9)%,(66.7±7.8)% ,P ＜ 0.01 and 0.05].CONCLUSION: It may increase the incidence rate of carcinoma of large intestine when pectin is added in diet, on the contrary, the rate decreases if inositol hexaphophate is added in the water.

Objective To detect the expression and function of ZNRF3 in different kinds of thyroid cancer tissues and cells.Methods Immunohistochemistry was used to detect the expressions of ZNRF3 protein in 35 cases of papillary thyroid carcinoma and 10 cases of poorly differentiated thyroid carcinoma.The expressions of ZNRF3 gene in TPC-1 and 8505C were detected by RT-PCR,and the cell lines were derived from papillary thyroid carcinoma and poorly differentiated thyroid carcinoma respectively.After silenced ZNRF3 gene expression with lentivirus,the proliferation ability of TPC-1 cells were detected with CCK-8,the invasion and metastasis ability of TPC-1 cells were detected with Transwell.Results According to results of immunohistochemistry and RT-PCR,the expressions of ZNRF3 in papillary thyroid carcinoma cells and tissues were higher than those in poorly differentiated thyroid carcinoma cells and tissues,the differences were statistically significant (4.83±0.44 vs.3.13 ±0.59,t =2.20,P ＜0.05;1.01±0.06 vs.0.21±0.04,t =11.80,P＜0.01).After ZNRF3 geng silencing,according to the results of CCK8,the proliferation ability of TPC-1 cells was significantly enhanced in 72 h,the difference was statistically significant (0.96 ± 0.10 vs.0.64 ± 0.05,t =3.19,P ＜ 0.05);and according to the results of Transwell,the TPC-1 cell's invasion (0.12 ± 0.01 vs.0.09 ±0.00,t =5.48,P＜0.01) and migration (0.22 ±-0.01 vs.0.17 ±0.01,t =4.58,P ＜0.05) also increased,the differences were statistically significant.Conclusion The expression of ZNRF3 in papillary thyroid carcinoma is higher than that in poorly differentiated thyroid cancer.ZNRF3 is tumor suppressor gene in the thyroid tumors.