目的:研究蛇葡萄素钠对人肺腺癌SPC-A-1细胞、人正常胚肺MRC-5细胞的毒性作用。方法:应用MTT法检测细胞增殖,流式细胞仪检测细胞周期和细胞凋亡;透射电子显微镜（TEM）观察细胞超微结构的变化。结果:MTT法实验表明,蛇葡萄素钠、卡铂显著抑制SPC-A-1细胞的增殖,呈现浓度依赖性,IC50分别为57.18±9.42 ug/ml、33.56±11.2 ug/ml,但对MRC-5细胞增殖的抑制作用不明显,IC50分别大于5000 ug/ml及500ug/ml;流式细胞检测分析表明,25~100ug/ml的蛇葡萄素钠可使SPC-A-1细胞增殖阻滞于S期,G0/G1期细胞明显减少,且呈浓度依赖性,50、100 ug/ml的蛇葡萄素钠可使SPC-A-1细胞发生明显凋亡,而蛇葡萄素钠对MRC-5细胞周期及凋亡的影响不明显。蛇葡萄素钠浓度为50、100ug/ml时,透射电镜下可见SPC-A-1细胞表现出典型的凋亡特征,而MRC-5细胞变化不明显。结论:蛇葡萄素钠对肿瘤细胞具有较强的毒作用,对正常细胞毒性较小。

OBJECTIVE To study the effects of vitamin D3 combined with metformin on the proliferation and apoptosis in human bladder cancer cell line SW-780 and its possible mechanism. METHODS MTT assay and fluorescence microscope observations were used to study the effects of vitamin D3 combined with metformin on the proliferation and apoptosis of SW-780 cells in vitro. Western blot was used to detect the expression of apoptosis-related proteins p-Bcl-2, Bax, Cyclin D1, c-Myc and related signaling pathways activated proteins p-IGF-IR, p-mTOR, p-P70S6K, p-S6. RESULTS MTT results showed that 320 μg/ml vitamin D3 combined with 620 μg/ml metformin acting on cells for 48h had a significant synergistic effect on proliferation. Fluorescence microscope observations showed that compared with negative control group and monotherapy treatment group, the apoptosis features of combination treatment group were obvious and the apoptosis rate increased greatly. Western blot showed that compared with the negative control group and monotherapy treatment group, the expression levels of p-Bcl-2, Cyclin D1 and c-Myc in combination treatment group significantly decreased, whereas the expression level of Bax significantly increased, and the expression levels of p-IGF-IR, p-mTOR, p-P70S6K and p-S6 in combination treatment group significantly decreased. CONCLUSION Vitamin D3 combined with metformin exhibited obvious inhibitory effects on the cell proliferation and apoptosis induction in SW-780 cells. The underlying anti-tumor mechanism might be related to inhibiting the expressions of p-Bcl-2, Cyclin D1, c-Myc, p-IGF-IR, p-mTOR, p-P70S6K, p-S6 and activating the expression of Bax.

Metformin is usually used for the treatment of type 2 diabetes. Recently, many studies suggest that metformin and vitamin D have broad-spectrum antitumor activities. Our aim in this research was to study the effects of vitamin D3 combined with metformin on the apoptosis induction and its mechanisms in the human breast cancer cell line MDA-MB-231. Cell proliferation was measured by methylthiazol tetrazolium (MTT) assay. The morphology of cell apoptosis was observed after Hoechst 33342 staining. Here we show that vitamin D3 280 μg/ml or vitamin D3 300 μg/ml or vitamin D3 320 μg/ml seperately combined with metformin 15000 μg/ml exhibited synergistic effects on cell proliferation and apoptosis. The underlying anti-tumor mechanisms may involve m-TOR related pathways, which are related to activating expression of cleaved caspase-3, Bax and p-AMPK, as well as inhibiting expressions of p-Bcl-2, c-Myc, p-IGF-IR, p-mTOR, p-P70S6K, p-S6.

A previous study reported that ginsenoside Rd (GRd) after oral administration has a valuable effect against 2,4,6-trinitrobenzenesulfonic acid (TNBS)-induced recurrent ulcerative colitis. In this study, we use HPLC method to investigate the oral bioavailablity of GRd and found that GRd was very poorly absorbed following oral administration in vivo. Simultaneously, the amount of GRd was degraded 71.77 +/- 4.8% in the gastric juice, the amount of GRd in the colon 6 h after oral administration was 19.8 +/- 13.7% of the given dose. In further experiment, we found 81.25 +/- 9.74% of the given dose of GRd in the colon of mice after duodenal administration. GRd was detected and stable in the intestine contents of mice. These pharmacokinetic characteristics suggested that GRd affected the ulcerative colitis directly other than its a few other metabolic products. So that, it has a natural colon-targeting feature and it may be possibly developed as a therapeutic agent to inflammatory bowel disease.

Ulcerative colitis (UC) is characterized by oxidative and nitrosative stress and neutrophil infiltration. In the present study, we aimed to investigate the therapeutic effect of ginsenoside Rd (GRd) in rats with 2,4,6-trinitrobenzenesulfonic acid (TNBS)-induced recurrent UC. After UC was twice-induced by intracolonic injection of TNBS, rats were intragastrically administered different doses of GRd per day for 7 days. The colonic lesions and inflammation were evaluated both histologically and biochemically. Compared with the TNBS group, GRd treatment facilitated recovery of pathologic changes in the colon after induction of recurrent UC, as evidenced by a significant reduction of colonic weight/length ratio and macroscopic and microscopic damage scores (p < 0.01). The myeloperoxidase and inducible nitric oxide synthase activities with malonyldialdehyde and nitric oxide levels in colonic tissues were significantly decreased in the GRd group compared with those in the TNBS group (p < 0.01). GRd treatment was associated with remarkably increased superoxide dismutase and glutathione peroxidase activities. Results showed a valuable effect of GRd against TNBS-induced recurrent UC by inhibiting neutrophil infiltration and promoting the antioxidant capacity of the damaged colonic tissue.

肤疡宁软膏为一临床验方,临床用于治疗皮肤烧伤疗效显著.该方是由油煎白芷、伏龙肝、冰片、白矾等组成、用麻油和蜂蜡制成的软膏剂.白芷具有解热、镇痛与抗炎作用[1],由天津第一中心医院配制的白芷糊软膏(白芷、紫草、冰片等)用于烧烫伤有较好的效果[2].冰片作为一个辛开之品,具有辛散透表之功,能引药由肌表直达腠理,提示冰片本身可以作为促渗剂应用于经皮给药制剂[3~4].白矾可清热毒、燥湿等,具有抗菌消炎、收敛的作用[5].肤疡宁软膏作为验方,其组方及工艺是否合理,尚待证实.从对祖国医学传统经验方挖掘、提高的思路出发,本实验研究了肤疡宁的处方工艺及其对小鼠皮肤烫伤的影响.

Objective To observe the effects of mouse nerve growth factor(NGF)on 2,5-hexanedione (2,5-HD)induced toxic peripheral neuropathy in rats.Methods Rats were administered different doses of NGF 1000,2 000,and 4 000 BU/(kg·d)intragastrically for 11 days after peripheral neuropathy had been induced by intermittent exposure to 2,5-HD for 37 days.Normal control group and model group were given the same amount of physiological saline.The scores of neuropathic signs(gesture,gait and muscular strength abnormality),electrophysiological in the hind limb and histopathological examinations of the sciatic nerves were evaluated.Results When compared with normal control group,rats in 2,5-HD induced group displayed neurobehavioral and electrophysiological responses significant deterioration but they were obviously ameliorated in NGF-treated rats.2,5-HD intoxication produced a histopathological change in tibial nerve fibers,including swelling,degeneration of axons,and myelin sheath swelling.Having been treated with NGF 2 000 or 4000 BU/(kg·d)for 11 days,the sheath swelling of tibial nerve fibers was obviously alleviated as compared with that in model group,and the morphology of axons was very close to that in normal rats.Conclusion NGF exerts a biologically significant neuroprotective effect against 2,5-HD induced peripheral neurotoxicity in rats.It provides evidence for clinical application of NGF to treat patients with n-hexane neuropathy.

Objective To investigate the effect of sodium ampelopsin(AMP-Na) at the different dosages on the nervous,respiratory and cardiovascular systems.Methods AMP-Na were given to rats iv to observe the changes on the nervous system with low(200 mg·kg-1),middle(260 mg·kg-1) and high(320 mg·kg-1) doses.And AMP-Na was given to the anesthetized dogs iv to observe the effect on the respiratory system and cardiovascular system with low(45 mg·kg-1),middle(82.2 mg·kg-1) and high(150 mg·kg-1)doses.Results Compared with pre-drug and NS groups,the nervous and respiratory systems have no statistics difference after giving AMP-Na iv with low,middle and high doses(20 times of the normal dose on a person),either the cardiovascular system at low doses.But at middle and high dose,AMP-Na can decrease blood pressure and heart rate,especially in diastolic blood pressure.The administration of high-dose group also decreased QRS interval,but the changes of other ECG have no statistic difference.Conclusion It is safe that AMP-Na can be used in normal dose,but more attentions should be paid for its effects on the cardiovascular system at high dose.

Objective To study the distribution and excretion of synthetic thymosin α1(sTα1) in mice and rats after intravenous(i.v.) injection.Methods The sTα1 concentrations in tissues at different time points and cumulative amounts of sTα1 in urine over different periods after i.v.injection of sTα1 into mice were measured by competitive ELISA.The cumulative amounts of sTα1 in bile over different periods after i.v.injection of sTα1 into rats were also measured by competitive ELISA.Results The concentration of sTα1 in the thymus gland was the highest(1 533.4±712.4 ng/g) at 15 min after i.v.injection of sTα1(1 mg/kg) into mice,followed by those in the liver,spleen,and kidney,while sTα1 concentration in the brain was relatively low.At 30 min after i.v.injection of sTα1 into mice,sTα1 concentration in the thymus gland was still the highest(25 583.4±6 244.7 ng/g),followed by those in the spleen,kidney,and liver.At 1 h after i.v.injection of sTα1 into mice,sTα1 concentration in the kidney was the highest(6 164.1±4 821.8 ng/g),and that in the thymus gland was(5 665.8±3 881.9) ng/g.At 2 h after i.v.injection of sTα1 into mice,sTα1 concentration in the kidney was the highest(1 408.0±1 252.7 ng/g),and that in the thymus gland was(306.4±268.6) ng/g,with trace amounts of sTα1 in the other tissues.The concentration of sTα1 in the brain was similar to that in blood plasma,indicating that sTα1 could pass through the blood-brain barrier.After i.v.injection of sTα1(1 mg/kg) into mice,the cumulative amount of sTα1 in urine over 12 h was 26.35% of administration dosage.There was no sTα1 excretion in bile after i.v.injection of sTα1(0.5 mg/kg) into rats.Conclusion After i.v.injection of sTα1 into mice,sTα1 is mainly distributed in the thymus gland and spleen and primarily excreted through the kidney,and can pass through the blood-brain barrier.

OBJECTIVE:To study pharmacokinetic property of synthetic thymosin α1(sTα1) in mice and rats with intravenous(i.v.) and subcutaneous(s.c.) administration. METHODS:45 mice(sTα1,1 mg·kg-1) and 3 rats(sTα1,0.5 mg·kg-1) were given relevant medicine via tail vein. Blood samples were collected from orbit vein and heart before and within 6 h after administration. Another 180 mice were randomly divided into high-dose,medium-dose and low-dose groups(sTα1,5,1,0.32 mg·kg-1) . Another 9 rats were randomly divided into high-dose,medium-dose and low-dose groups(sTα1,2.5,0.5,0.16 mg·kg-1) . Six groups were given relevant medicine subcutaneously. Blood samples of orbit vein and heart were collected before and within 10 h after administration. ELISA was used to determine blood concentration of drugs at different time points and pharmacokinetic parameters were calculated. RESULTS:The main pharmacokinetic parameters of sTα1 in mice vs. rats after intravenous administration were as follows:t1/2β:0.68 h and AUC0～∞:554.32 μg·h·L-1;t1/2β and AUC0～∞ of sTα1 in rats after intravenous administration were(1.87±0.50) h and(1 602.91±360.41) μg·h·L-1. Pharmacokinetic parameters of sTα1 in mice in sTα1 high-dose,medium-dose and low-dose groups after subcutaneous administration were as follows:t1/2:0.76 h,0.54 h,0.268 h;AUC0～∞:3 222.95 μg·h·L-1,417.67 μg·h·L-1,366.60 μg·h·L-1,respectively. Pharmacokinetic parameters of sTα1 in rats in sTα1 high-dose,medium-dose and low-dose groups after subcutaneous administration were as follows:t1/2:(1.23±0.23) h,(1.40±0.37) h,(1.99±0.94) h;AUC0～∞:(22 436.74± 5 641.94) μg·h·L-1,(1 539.63±203.30) μg·h·L-1,(729.60±320.0) μg·h·L-1,respectively. CONCLUSION:Pharmacokinetic model of sTα1 in rats and mice with intravenous administration is first-order two-compartment model while that of sTα1 in rats and mice with subcutaneous administration is first-order one-compartment model.

The compound rh-IFNalpha-2a-NGR can inhibit tumor angiogenesis and could be used for targeted therapy. In the present study, double antibody sandwich ELISA analysis was used to determine the concentration of rh-IFNalpha-2a-NGR in serum after intramuscular administration of various dosages to mice, rats and monkeys. The results showed that the pharmacokinetic properties of rh-IFNalpha2a-NGR after i.m. administration to mice, rats and monkeys were consistent with a one-compartment open model. The main pharmacokinetic parameters in mice (9.36 microg/kg), rats (4.68 microg/kg) and monkeys (2.34 microg/kg) after i.m. rh-IFNalpha2a-NGR were as follows: T(peak) was 0.49, 1.65 and 3.60h, C(max) was 3030.20, 654.49 and 268.13 ng/L, t1/2 was 0.39, 4.52 and 2.70 h, and AUC(0-infinity)) was 4197.65, 5784.58 and 2622.06 ng/L x h, respectively. Also, mice, rats and monkeys had their own distinct metabolic characteristics. These data would provide references for further clinical pharmacokinetic study of rh-IFNalpha2a-NGR.

To investigate the protective effect of 1-(2 ,6-dimethylphenoxy)-2-(3, 4-dime-thoxyphenylethylamino) propane hydrochloride (DDPH) on myocardial ischemia-reperfusion (I/ R) injury in isolated rat hearts. METHODS: Myocardial ischemia-reperfusion injury models were built with Langendorff isolated perfusion technology and established by the ligation of left descending coronary artery (LAD) for 40 min and reperfusion for 120 min in isolated rat hearts. The influence of DDPH on left ventricular function, myocardial infarct size, lipid perox-idation and the ultramicrostructural changes of myocardial cells were observed. RESULTS: DDPH improved the damage of left ventricular systolic anddiastolicfunction causedbyI/R,obviously reduced myocardial infarct size, increased the activities of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px), decreased the content of MDA in myocardium, and lessen the degree of ultramicrostructural injury in myocardial cells. CONCLUSION: DDPH has a protective effect on myocardial ischemia-reperfusion injury in isolated rat heart, which may be related to inhibiting the formation of the oxygen free radical and subsequent lipid peroxi-dation.

Objective: To evaluate the immunogenicity and long-term hematological toxicity of Ⅲ valence rotavirus gene reassortment vaccine in Sprague-Dawley(SD) rats.Methods: Rats(n=30) were repeatedly ig immunized with Ⅲ valence rotavirus gene reassortment vaccine at 2 mL for 3 times.Three weeks after the first immunization,and 1 and 4 weeks after the third immunization,10 rats were respectively sacrificed for examination of the specific antibody against vaccine,hematology and bone marrow histopathology.Results: The antibody titers were significantly higher in vaccine group than control group at each detected point.The antibody against G1,G2,G3,G4 and G10 were found three weeks after the first immunization,and then increased gradually.The antibody titers of G2,G3 and G4 were significantly higher than G1 and G10(P0.01 or P0.05).The lymphocyte count and mean corpuscular hemoglobin(MCH) in vaccine group showed reversible changes,but did not show apparent toxicity.There were no apparent changes in bone marrow histopathology.Conclusion: Ⅲ valence rotavirus gene reassortment vaccine has immunogenicity against rats and has no apparent bone marrow toxicity in rats.

Metabolic syndrome (MS) is highly prevalent in developed countries and becoming a serious worldwide public health issue. In this study, we established a MS model by feeding male C57BL/6J mice with a high-fat diet (10%) for 18.5 weeks, studied the therapeutic effects of a recombinant mutant of the human ciliary neurotrophic factor (rhmCNTF) 0.1 (C-0.1) or 0.3 (C-0.3) mg x kg(-1) per day subcutaneously or pair feeding (PF, which mice were restricted to the same amount of food as eaten by C-0.3 treated mice) in MS mice. After 10 days treatment, rhmCNTF reduced obesity related indices, ameliorated glucose and lipid metabolism abnormality, and enhanced insulin sensitivity. In addition, liver function and antioxidant ability of MS mice were improved by rhmCNTF. Pair feeding revealed the same effects as C-0.3 on obesity related indices and insulin sensitivity, but aggravated hepatic steatosis and hepatic function. The results suggest that rhmCNTF could serve as an effective therapeutic agent for MS and related diseases.

AIM:To screen the hypoglycemic activity of diferent sulphonylureas derivative ligand and their metal complexes. METHODS:The mice were administered intragastrically,the blood samples were obtained from tail vein. The serum glucose levels were determined by glucose-oxidase-peroxidase (GOD-PAP) method. The hypoglycemic activities were analyzed by the mean of tolbutamide as a positive control drug,and its mechanism and structure-activity relationship between chemical structure and hypoglycemic activity were studied. RESUITS:The results showed that ZnL2·2H2O,NdL3·2H2O,SmL3·2H2O,Zn(HL')2·2NO3,Eu(HL')3·3NO3,HL and HL' all reduced the blood sugar in normal mice,while EuL3·2H2O,Nd(HL')3·3NO3 and Sm(HL')3·3NO3 had no obvious hypoglycemic activity. CONCLUSION:The hypoglycemic activity of HL is stronger than that of tolbutamide in normal mice at 2 h,indicating that when the first substituent of tosylurea is the alicyclic group (cyclohexyl),its hypoglycemic activity may be higher than the straight-chain lipid group (butyl),and much higher than the heterocyclic group (4-antipyrine).

OBJECTIVE: To investigate the long-term toxicity of repeated intramuscular injection of Escherichia coli O157 : H7 polysaccharide-conjugated vaccine(O157) in rats so as to provide safety evidence for clinical trials. METHODS: A total of 48 SD rats were randomly assigned to receive either 0.5 mL vaccine(containing 25 μg polysaccharides) (immunization group, n = 24) or phosphate buffered solution (PBS,control group, n = 24) with the same volume for 3 times at a dose interval of two weeks. Sacrifice of 6 rats in each group were scheduled at 2 weeks after first immunization, and at 1, 3, and 5 weeks after the third immunization, respectively for observation and determination of hematological and biochemical parameters, histopathology, specific antibody, myeloid tissue, the tissues in injection sites, etc. RESULTS: Compared with control group, immunization group showed no significant pathological change except the dynamic regular change of some hematological pa-rameters induced by the immunization, and no immunologic system damage was observed. CONCLUSION: Repeated intra-muscular injection with O157 vaccine in rats wouldn't cause overt toxicity and local irritation.

Objective: To investigate the effect of nerve growth factor (NGF) on acrylamide-intoxicated peripheral neuropathy in rats. Methods: Peripheral neuropathy was induced by intragastric administration of acryl-amide. The intoxicated rats were intramuscularly injected with NGF (1 000, 2 000 and 4 000 BU·kg~(-1), respective-ly) daily for 8 days. Neurobehavioral responses (landing foot splay and gait scores) were measured during the course of treatment with NGF. Electrophysiological test ( nerve-muscle action potential and nerve conduction veloci-ty) and histopathological examinations were performed on the next day after the last dose of NGF. Results: Acryl- amide significantly deteriorated neurobehavioral and electrophysiological responses, and caused an alteration of his- tomorphology in tibial nerve fiber (such as myelin sheath swelling and axons degeneration). NGF (1 000, 2 000 or 4 000 BU·kg~(-1)) attenuated these injuries, and reversed axons degeneration. Conclusion: NGF may be a useful treatment agent for patients with acrylamide-intoxicated peripheral neuropathy.

Objective:To evaluate the long-term toxicity of intramuscularly injected paratyphoid A polysaccharide protein binding vaccine in SD rats.Methods: Rats(n=24) were immunized by intramuscular injection of paratyphoid A polysaccharide protein binding vaccine at 25 μg per rat for 3 times.Saline-glycine solution 0.5 mL was injected as control(n=24).At 2 weeks after the first immunization,and 1,3 and 5 weeks after the third immunization,6 rats were respectively sacrificed for examination of hematology,serum biochemistry,the specific antibody against vaccine,and histopathology.Results: In rats immunized with the vaccine,reversible changes of hematology were observed,but did not show apparent toxicity.Immunotoxic damage was not found by histopathology examination in specific antibody-positive rats,suggesting that the specific antibody would not be harmful to the rats.There were no significant differences in the indices of haematology and biochemistry between vaccine group and control group.Conclusion: Paratyphoid A polysaccharide protein binding vaccine has potent immunogenicity,but no detectable toxicity after repeated immunization in rats.

目的:研究柴黄片的解热作用。方法:应用干酵母及角叉菜胶致大鼠发热的模型研究柴黄片的解热作用。结果:3、6、12g/kg ig柴黄片均可以降低由干酵母及角叉菜胶所致的大鼠发热作用,大鼠体温呈逐渐下降趋势,其解热作用呈剂量依赖性。结论:柴黄片有解热作用。

Objective:To study the protective effects of Salvia przewalskii Maxim granule(SPG)on experimental myocardial ischemia and coagulation system.Methods:The acute myocardial ischemia model was established on rats by ligation of coronary artery;meanwhile,the changes of S-T segment,myocardial infract size,contents of LDH and myocardial function were determined and assessed.The myocardial ischemia model was made by isoproterenol on mice and the contents of LDH and MDA were determined.The coronary flow of rats was observed by langendorff's method.The survival rate of mice was observed after thrombus was formed induced by collagen and epinephrine.The platelet aggregation was determined by Born's method.Results:SPG can significantly reduce myocardial ischemia S-T segment elevation,the degree of myocardial ischemia and the myocardial infract size,decrease the contents of LDH and MDA of rats and mice,improve the contraction function of rats with myocardial ischemia and increase the flow of coronary.SPG has a significant inhibiting effect on the platelet aggregation of rabbits,and can improve the survival rate of mice which have formed thrombus induced by collagen and epinephrine.Conclusion:SPG has protective and therapeutic effects on the experiental myocardial ischemia of rats and mice,and it can inhibit platelet aggregation of rabbits and thrombus formation of rats.