AIM: To investigate the anti - tumor effect of Weimaining (WMN) on a murine Lewis lung carcinoma (3LL) and the influence on the cell cycle. METHODS: The inhibitory rate of WMN in 3LL growth was detected by replicating the model of 3LL. The effect of the drug on 3LL cell cycle and the influence of the drug on the expression of cy clin D1 protein were investigated by flow cytometry and immunohistochemical staining. RESULTS: The results showed that the inhibitory rate of drug in 3LL is 19. 14%, 33.59%, 40. 63% and 51.56% respectively at dosage ranging from 100,cells in G0 -G1 phase and decreases the expression of cyclin D1 protein. CONCLUSION: WMN inhibits the growth of 3LL cells in vivo by decreasing the expression of cyclin D1, blocking the cells in G0 - G1 phase and preventing the cells transition from G1 to S phase while DNA is replicated.

Objective:To study the effects of Yiqi Huoxue Ruanjian Jiedu(YHRJ) drug serum on Ca 2+ and mitochondrial membrane potential (△?m) in human hepatocellular carcinoma cell line BEL-7402.Methods: YHRJ decoction was composed of Astragalus Root, White Atractylodes Rhizome, Radix Notoginseng, Fructus Polygoni Orientalis, Radix Actindiae Argutae, Flos Campsis, Pangolin Scales, Oldenlandia diffusa Roxb, and Herba Scutellariae Barbatae. Twelve New Zealand rabbits were randomly assigned into 3 groups: NS group, YHRJ equivalent dose group(YHRJD) and YHRJ high dose group(YHRJG), which were fed with NS,YHRJ equivalent dose decoction (5 times as large as normal human dose) and high dose YHRJ decoction(10 times as large as normal human dosage), respectively. Drug serum was prepared after the rabbits were fed with drugs for 5 times. BEL-7402 cells were divided into 6 groups according to different drugs, including NS group, NS+DDP group, YHRJ equivalent dose group, YHRJ equivalent dose+DDP group, YHRJ high dose group, and YHRJ high dose +DDP group. The concentrations of NS,YHRJD and YHRJG were adjusted to 100 ml/L ,and the concentration of DDP was adjusted to 4.5 mg/L. Flow cytometry was applied to detect the concentrations of Ca 2+ and △?m 8, 12, 24 and 48 h after the drugs were added.Results: The Ca 2+ concentrations at 12, 24 and 48 h after treatment were significantly higher than those of the control group(P

Objective To investigate the effects of curcumin on H2O2-induced the production of nitric oxide(NO) and reactive oxygen species(ROS) in mouse embryo-fibroblast.Methods Macrophage were collected in abdominal cavity of 6-8 weeks Kunming mouse,cultured macrophage(2?108 L-1) were divided into control group and curcumin groups randomly.Macrophage in H2O2 group were added into a single bolus of H2O2(1 mmol?L-1) for 30 min,macrophage in curcumin group were preincubated with different concentration of curcumin for 2 h followed by a 30 min incubation with 1 mmol?L-1 H2O2 and macrophage in control group were added into the same volume(0.1 mL) of 9 g?L-1 so-dium chloride.Immunocytochemistry was used to measure the contents of inducible nitric-oxide synthase(iNOS),NO production was determined by Griess reactive,ROS was determined by DCFH-DA Fluorescence proe.Results The production of NO in control group was little.The production of NO in H2O2 group markly highter than that in control group(P

OBJECTIVETo study the medium and long term effects of perfusion of bone marrow stromal stem cells through optional artery for the treatment of non-traumatic femoral head avascular necrosis.

METHODSFrom January 2000 to December 2004,62 cases(78 hips) with non-traumatic femoral head necrosis accepted optional artery marrow stromal stem cells infusion treatment and had complete follow-up data, including 43 hips of 35 males and 35 hips of 27 females with an average age of 36.3 years old (22 to 54). According to preoperative imaging data, 16 hips were ARCO I stage, 52 hips were II stage, 10 hips were III a stage. Harris score was 64.94 +/- 8.12 preoperatively. Postoperative Harris score at the last follow-up, imaging changes,DSA vascular changes were analysis.

RESULTSThe patients were followed up for 9 to 13 years (means 11 years). By the end of the follow-up, a total of 18 hips got artificial joint replacement, 10 hips of preoperative ARCO I, II period got artificial hip joint replacement, 8 hips of IIIa period got hip artificial joint replacement. Harris score was 71.21 +/- 0.19 at the end of the follow-up, it was obviously enhanced compared with preoperative. DSA showed blood vessels of supply the femoral head increased thickening.

CONCLUSIONPerfusion of bone marrow stromal stem cells through optional artery can effective treat non-traumatic femoral head necrosis of ARCO I, II period, it can make the femoral circumflex artery and its branches increased thickening.

Adult ; Angiography, Digital Subtraction ; Arthroplasty, Replacement, Hip ; Female ; Femur Head Necrosis ; therapy ; Follow-Up Studies ; Humans ; Male ; Mesenchymal Stem Cell Transplantation ; Middle Aged

Streptomyces sp. A048 was cultured in a complete medium to the last stage of log phase,the hyphae were washed and collected by centrifugation. Then the hyphae were inoculated in liquid medium for chitinase production using two-step fermentation. Activity of chitinase produced by two-step fermentation was 1.1 times higher than that from one-step fermentation,and ferment cycle was for 54 hours,which was 66 hours shorter than that of one-step fermentation. The hyphae and the powder of chitin were co-immobilizated and cultured in liquid medium for 36 hours,activity of chitinase was 1.8 times higher than that from one-step fermentation,and ferment cycle was 54h shorter than that of one-step fermentation. By adding 0.4% cellulose to two-step fermentation,activity of chitinase was 18.52 U/mL that was 4 times higher than that from the control and 10 times higher than that from one-step fermentation. Two step fermentation with chitin and cellulose may be the optimal fermentation process to produce Chitinase from Streptomyces sp. A048.

AIM: To investigate the anti-metastasis effect of weimaining, extracted from fragopyrum cymosum meissn, a Chinese medicine, on murine Lewis lung carcinoma (3LL). METHODS: The anti-metastasis effect of weimaining in vivo was detected in the grafting lung metastasis model of murine Lewis lung carcinoma. The effects of the drug on the expression of CD34 and E-cadherin were investigated by immunohistochemical staining and RT-PCR. RESULTS: Weimaining effectively inhibited the lung metastasis of 3LL at a concentration of 250 mg?kg -1?d -1, significantly suppressed the expression of CD34 and increased the expression levels of E-cadherin protein and mRNA in 3LL cells. CONCLUSIONS: Weimaining inhibits the metastasis of murine Lewis lung carcinoma (3LL) in vivo via increasing the expression of E-cadherin and decreasing microvessel density of tumor tissue.

AIM: To investigate the effects of Chinese medicine, Jinan injection, on ultrastructure and mitochondria in cultured lung cancer cell lines. METHODS: The cultured lung cancer cell lines PG and PAa were used and divided into 4 groups: control (C), cisplatin (DDP), Jinan (JA) and Jinan in combination with cisplatin (DJ), respectively. The changes of morphology and mitochondria membrane potential, intracellular Ca~(2+) and pH in every group were observed by inverted microscope and electronic microscope as well as by using flow cytometry, staining by rhodamine, Fluo-3 and BCECF, respectively. RESULTS: Degeneration cells showed chromatin condensation and peripheral congregation. In cytoplasm autophage lysosome increased and myelinoid body was seen easily. In mitochondria structure, where the space between the inner and outer membranes of these organelles expanded as the matrix was compressed. The electron-dense or swelled was observed as vacuole degeneration and its matrix showed electron-lucent. Compared to control, mitochondria membrane potential increased in every group after 24 h and 48 h treatment. DDP increased intracellular calcium ion in PG cells, however, in PAa cells, JA and DJ decreased it. Intracellular pH got lower at 24 h and higher at 48 h in PG and PAa cells. There were significance in every group vs control in PG and PAa by statistic t-test (P

Objective To investigate the effects of silybin-phosphatidylcholine compound (SPC) on H2O2-induced the production of nitric oxide (NO) and reactive oxygen species (ROS) in mouse macrophage.Methods Macrophage were collected in abdominal cavity of 6-8 week Kunming mouse,and cultured macrophage(2?108 L-1) were divided into control and SPC group randomly.Macrophage in control group were added into the same volume(0.1 mL) of 9 g/L sodium chloride.Macrophage in H2O2 group were added into a single bolus of H2O2 (1 mmol/L H2O2) for 30 min,while macrophage in SPC group were preincubated with different concentration of SPC for 2 h followed by a 30 min incubation with 1 mmol/L H2O2.Immunocytochemistry were used to measure the contents of inducible nitric-oxide synthase(iNOS),NO production was determined by Griess reactive, ROS was determined by DCFH-DA Fluorescence probe.Results The production of NO in H2O2 group markedly higher than that in control group(P

Interleukin-1 receptor antagonist (IL-1ra), a member of IL-1 family, is a naturally occurring IL-1 inhibitor as "receptor antagonist", which blocks biological responses mediated by IL-1. Recombinant human IL-1ra (rhIL-1ra, Kineret) was introduced in clinical trials involving patients with RA. Between 2001 to approximately 2002, rhIL-1 ra was approved by the US Food and Drug Administration and the European Agency for the Evaluation of Medicine Procedure. Unfortunately, 10,000 to 100,000-fold excess amounts of IL-1ra are needed to relieve disease because minimal IL-1 can induce complete biological responses, and the dosage of 100 to approximately 150mg/day in a RA patient is so big that it greatly influence patients' physical, psychological and economical situation. In this study, IL-1ra mutants were established by site-specific mutagenesis to improve its stability. The sites of mutagenesis included R6 K7-AA,R93 K94-AA and K97 R98-AA. IL-1ra and its mutants were expressed in E. coli BL21 (DE3) using pTIG-Trx expressing system with the induction of IPTG. The recombinant proteins were purified by Ni2+ chelate chromatography and Sephadex G75 gel filtration chromatography. The activity of mutants is as high as IL-1ra. We characterized the pharmacokinetic profile of IL-1ra and its mutants. The third mutant's half life is 2.26 times than wt IL-1ra. The study has provided some approaches and experience for further research to improve the metabolism stability of IL-1ra.
Animals ; Escherichia coli ; genetics ; metabolism ; Female ; Humans ; Interleukin 1 Receptor Antagonist Protein ; biosynthesis ; genetics ; pharmacokinetics ; Mutagenesis, Site-Directed ; methods ; Mutant Proteins ; biosynthesis ; pharmacokinetics ; Recombinant Proteins ; biosynthesis ; genetics ; pharmacokinetics

Objective To explore the compound matrine injection on human hepatoma smmc-7721 cells' proliferation.Methods Human hepatoma smmc-7721 cells were sterilely cultured.Assayed the effects of compound matrine injection on smmc-7721 cells by MTT.The concentrations of the compound matrine injection were 3.34,2.38,1.43 and 0.48 mg/ml; Used the double staining method of annexin V-fluorescein isothiocyanate(Annexin V-FITC)/propidium iodide (propidium iodide,PI) to measure the apoptosis of hepatoma cell.The concentrations of the compound matrine injection were 0.86,1.72,and 3.43 mg/ml.Results The apoptosis rates of compound matrine injection with different concentration of 3.34,2.38,1.43 and 0.48 mg/ml on liver cancer cell SMMC-7721 were51.03％、53.67％、49.83％ and 0.03％ respectively after 24 hours; and 67.14％、65.35％、62.25％ and 0.05％ respectively after 48 hours;and 74.16％、68.77％、66.04％ and 26.02％respectively after 72 hours.The apoptosis rates of compound matrine injection with different concentration of 0.86,1.72 and 3.43 mg/ml on liver cancer cell SMMC-7721 were(2.86±0.35)％,(15.16± 1.15)％and(13.17±0.40)％ respectively after 24 hours; and (8.57±0.44)％,(28.07±0.76)％ and (29.81±8.10)％respectively after 48 hours,which were significantly higher than the control group (1.05±0.09)％(P＜0.05).Conclusion The compound matrine injection has a significant inhibition on human hepatoma smmc-7721 cells' proliferation and the mechanism was its inducing cell apoptosis.