AIM: To investigate the influences of nimodipine on HCT cells proliferation and explore the mechanism. METHODS: HCT cells were treated with different concentrations of nimodipine, and its proliferation was inspected by MTT assay. The apex areas of sub diploid were measured by Flow Cytometry and the DNA ladder were found by agarose gel electrophoresis. The characteristic changes in morphology were observed under the light microscopy. The cellular distribution and concentration of calcium were studied by using the laser confocus scanning microscopy. RESULTS: The growth of HCT cells was inhibited by different concentrations of nimodipine. Data from Flow Cytometry showed the apex areas of sub diploid enlarge in the drug treating group, suggesting that the number of apoptosis cells increased in dose dependent manner. Gel electrophoresis displayed DNA cleavage pattern typical of apoptosis: DNA ladder in high dose nimodipine treated HCT cells. The light microscopy presented cellular morphological changes: cell membrane blebs, the cytoplasm and nuclear chromatin condensation. The concentration of cytosolic free calcium increased when treated with nimodipine showed by the laser confocus scanning microscopy. CONCLUSION: Nimodipine can cause inhibition of the cell proliferation of HCT cells. The mechanism of inhibition might involve the cell apoptosis.

AIM To elucidate whether apoptosis is involved in the development of multidrug resistance in 5-fluorouracil resistant cells Bel7402/5-FU. METHODS Multidrug resistance of Bel7402/5-FU cells was determined by SRB assay, apoptosis was measured by Annexin V-FITC/PI double labeled assay, cell cycle phases were analysed by flow cytometry, and expression of genes related to apoptosis bcl-2, bcl-xl, bcl-xs, bax, fas, p53, and cpp32 was detected by RT-PCR assay. RESULTS Bel7402/5-FU cells were resistant not only to 5-FU but also to multiple anticancer drugs. When treated with the same concentration of 5-FU (30 ?mol?L -1 or 150 ?mol?L -1 ), the proportion of apoptosis was significantly increased and G_0/G_1 phase was increased and S and G_2/M phases were reduced in Bel7402 cells, but the proportion of apoptosis and cell cycle was not changed in Bel7402/5-FU cells. The expression of bcl-xl, bcl-xs, and bax mRNA were significantly increased and the expression of p53 and cpp32 mRNA were significantly decreased in resistant Bel7402/5-FU cells, as compared with Bel7402 cells. CONCLUSION Decrease of apoptosis may be related to multidrug resistance of human hepatocellular carcinoma cells Bel7402/5-FU. The up-regulation of anti-apoptotic genes and down-regulation of pro-apoptotic genes are involved in the development of multidrug resistance in Bel7402/5-FU cells.

AIM: To develop a simple, convenience, and inexpensive method on primary culture model of pancreatic islets in rats for the study of anti-diabetic drugs. METHODS: The pancreases of SD rats were separated from the pancreatic duct with cold Hank' s solution and picked. Then the pancreases were cut into pieces and repeatedly digested by collagenase at 37℃ for the short durations of the experiment. The isolated islets were identified by dithizone staining and the viability was evaluated by trypan blue staining. Pancreatic islets were incubated in RPMI 1640 or DMEM for 14 - 16 h, then they were transferred to new culture plates with the same medium mentioned above. Determination of insulin content of su-pernate and cell lysate and the experiment of insulin secretion by stimulation of glucose and implantation of micewith STZ-induced diabetes were used for evaluated the function of islets. RESULTS: The viability of isolated pancreatic islets was more than 95% and the purity of cultured islets was about 85% . The insulin synthesis, secretion and sensitivity of islets stimulate by glucose which were cultured in RPMI 1640 were higher than that in DMEM. The levels of blood glucose recovered to normal in type 1 diabetic mice after islets implantation. CONCLUSION : The islets got in this study have higher purity and viability with the normal biological activity for about 7 days by this method and they can be used as a cell model for the study of diabetes in vitro .

AIM: To develop a reliable primary cultured hepatocytes model in vitro for liver metastasis research. METHODS: Hepatocytes were isolated by a modification of the two-step collagenase perfusion method. The apoptosis and cell cycle of hepatocytes were measured with flow cytometry. The proliferation of hepatocytes was detected by SRB method. RESULTS: The viability and purity of hepatocytes were 90% and 95%,respectively. The result of flow cytometry analysis showed that there was little apoptosis in hepatocytes and most of hepatocytes were in G_0/G_1 phase. The proliferation and albumin-secreting function of hepatocyte cultured by low glucose DMEM and high glucose DMEM were higher than that of cultured by RPMI1640 during 1 to 6 day, but there was no significant different between low glucose DMEM group and high glucose DMEM group. CONCLUSION: Hepatocytes have higher purity and viability with the normal biological activity for about 6 days by this method and it may be a cell model for the study of liver metastasis in vitro.

The anti-inflammatory effect of total saponins of Panax notoginseng (TSPR ) were reported. After intramuscular injection of TSPR (200 mg/kg, rats and mice) and hydrocortisone ( 300mg/kg, rats ; 200mg/ kg, mice) the ear inflammation induced by croton oil decreased. Thirty minutes after intramuscular injection of TSPR ( 200mg/kg ) and hydrocortisone ( 200mg/kg ) in normal mice, the inflammation induced by acetici ( ip) shown by the infiltration of Even's blue decreased significantly.It is suggested antiinflammatory effect of TSPR was not related to the hypothalamic adrenal system.In chronic inflammatory experiments, TSPR as well as hydrocortisone could inhibit the proliferation of granuloma by the implantation of cotton pellet in rats and mice with inhibitory rates of 27.6% ( P

AIM: To detect sequence and mutation of K-ras oncogene in tissue and stool DNA of patients with colorectal cancer in order to provide a method of noninvasive and simple colorectal cancer diagnosis. METHODS: DNA was separated and purified from colorectal cancer tissue or stool of patient with colorectal cancer, then the K-ras gene was amplified by PCR and PCR products were cloned, the K-ras gene was sequenced, and the mutation was identified. The expression of color/colorectal cancer antigen was inspected by immunohistochemical technique. Stool sample of patient with colorectal cancer was detected with enzyme-linked immunosorbent assay (ELISA). RESULTS: K-ras gene sequence of the stool was completely same as that of the tissue of the patient;K-ras mutation was detected in one case. There was relativity between the mutation of K-ras gene and the pathology type of colorectal cancer and the expression level of colorectal cancer antigen in stool sample. CONCLUSION: It is feasible that colorectal tumors can be detected by a noninvasive method based on the molecular pathogenesis of the disease. Detecting K-ras gene mutations of stool DNA can provide bases for the screening, early detection, and prognosis to patients with colorectal cancer.

AIM: To study the influence of injection agent of pidotimod on function of the immune-system in normal and immune-depressed mice, and to investigate the mechanisms of the immunomodulating action of pidotimod. METHODS: The expressions of TNF-? and IL-6 in mice spleen was detected by RT-PCR method. The effects of pidotimod on phagocytosis functions of mice peritoneal exudate cells was investigated by neutral red phagocytosis. The lymphocyte proliferations induced by Con A or LPS were detected by MTT method. Adopt the serum haemolysis to measure the production of serum antibody. RESULTS: The normal and immune-depressed mice were treated for 14 days with different dosages of pidotimod by injection. Pidotimod can significantly increase the expressions of TNF-? and IL-6 of spleen, potentiate phagocytosis of the peritoneal exudate cells, potentiate the lymphocyte proliferations ability induced by Con A or LPS. CONCLUSION: Pidotimod can potentiate amelioration normal and immune-depressed mice immunesystem function and increase expressions of TNF-? and IL-6 of spleen.

OBJECTIVETo compare the antitussive activity of three kinds of Stemonae Radix specified in the Chinese Pharmacopoeia, including roots of Stemona sessilifolia, S. japonica and S. tuberosa.

METHODThe antitussive activity was determined in mouse after cough induction by ammonia aerosol stimulation and the number of cough in 2 min were detected with codeine as positive control.

RESULTAll the decoctions, the total alkaloid fractions and non-alkaloid fractions of S. sessilifolia, S. japonica and three chemical types of S. tuberosa showed significant antitussive effect and exhibited a dose-dependent inhibition of coughing. The ED50 values showed that the antitussive activity strength for both total alkaloid fractions and the decoctions are: S. tuberosa (Type I) > S. sessilifolia > S. japonica. The total alkaloid fractions had more potent atitussive activity than the decoctions and non-alkaloid fractions. The antitussive activity strength for the three chemical types of S. tuberosa is: Type I > Type III > Type II. The samples from different producing areas for the same species of Stemonae Radix had no significant differences in antitussive activity. The result also showed that the honey-processed slice had much stronger antitussive activity than raw slice.

CONCLUSIONThe antitussive efficacies of Stemonae Radix were influenced by chemical diversity both in same species and among different species, different fractions and processed method.

Animals ; Antitussive Agents ; administration & dosage ; Cough ; drug therapy ; Disease Models, Animal ; Drugs, Chinese Herbal ; administration & dosage ; Humans ; Male ; Mice ; Mice, Inbred ICR ; Stemonaceae ; chemistry

Metabolic reprogramming is a hallmark of cancer, including lung cancer. However, the exact underlying mechanism and therapeutic potential are largely unknown. Here we report that protein arginine methyltransferase 6 (PRMT6) is highly expressed in lung cancer and is required for cell metabolism, tumorigenicity, and cisplatin response of lung cancer. PRMT6 regulated the oxidative pentose phosphate pathway (PPP) flux and glycolysis pathway in human lung cancer by increasing the activity of 6-phospho-gluconate dehydrogenase (6PGD) and α-enolase (ENO1). Furthermore, PRMT6 methylated R324 of 6PGD to enhancing its activity; while methylation at R9 and R372 of ENO1 promotes formation of active ENO1 dimers and 2-phosphoglycerate (2-PG) binding to ENO1, respectively. Lastly, targeting PRMT6 blocked the oxidative PPP flux, glycolysis pathway, and tumor growth, as well as enhanced the anti-tumor effects of cisplatin in lung cancer. Together, this study demonstrates that PRMT6 acts as a post-translational modification (PTM) regulator of glucose metabolism, which leads to the pathogenesis of lung cancer. It was proven that the PRMT6-6PGD/ENO1 regulatory axis is an important determinant of carcinogenesis and may become a promising cancer therapeutic strategy.