Callus of Gentiana crassicaulis was initiated from hypocotyl and cotyledon on MS medium supplemented with 2mg/L 2.4-D and 0.5 mg/L BA.Induction frequencies of callus were 100%.Somatic embryos were formed on MS medium containing 2mg/L BA,3mg/L ZT, 1mg/L NAA,3mg/L GA,three time MS organics and vitamins,6% sucrose and 500 mg/L LH and three time FeSO4 (Na2EDTA) .40% Callus produced somatic embryos.It developed into complete plants on hormon-free MS medium.

Objective To investigate the curative effects and adverse reactions of pure radiotherapy and concurrent chemotherapy and radiotherapy for patients with cervical cancer.Methods One hundred and twenty-seven patients with cervical cancer who accepted treatment in the Affliated Hospital of Inner Mongolia Medical Unive.rsity from May 2010 to May 2012 were collected.All patients were divided into two groups:pure radiotherapy group (n =65) and concurrent chemotherapy and radiotherapy group (n =62).The curative effects,adverse reactions and survival of two groups were observed.Results All patients were completed treatment.The median follow-up time was 42 months.The rate of complete response in the pure radiotherapy group was 80.0％ (52/65),and the rate in the concurrent chemotherapy and radiotherapy group was 82.26％ (51/62),with no significant difference (x2 =1.22,P =0.352).The 1-year overall survival rates in the pure radiotherapy group and the concurrent chemotherapy and radiotherapy group were 95.38％ and 95.16％ respectively,with no significant difference (x2 =0.32,P =0.533),but the 3-year overall survival rates were 81.54％ and 90.32％ respectively,the 5-year overall survival rates were 72.31％ and 83.87％ respectively,with significant differences (x2 =5.09,P =0.015;x2=3.87,P =0.039).However,for the patients who were ≥ 60 years,the 1-year overall survival rates in the two groups were 94.62％ and 93.91％ respectively,the 3-year overall survival rates were 85.02％ and 87.25％ respectively,the 5-year overall survival rates were 70.06％ and 73.58％ respectively,with no significant differences (x2 =0.06,P =0.753;x2 =1.16,P =0.279;x2 =0.48,P =0.511).The adverse reactions were mainly in grades 1-2.There were significant differences in the rates of leucopenia (56.10％ vs.72.20％),thrombocytopenia (58.82％ vs.76.80％),nausea and vomiting (34.04％ vs.56.90％) among the two treatment groups (x2 =11.23,P =0.003;x2 =11.82,P=0.002;x2 =12.77,P =0.000).Conclusion The curative effect of concurrent chemotherapy and radiotherapy is better than that with pure radiotherapy for patients with cervical cancer,which can improve the 3-year and 5-year overall survival.But at the same time,it should be noted that the rates of adverse reactions may be increased during the same period.For the age of 60 or more patients with cervical cancer,concurrent chemotherapy and radiotherapy does not achieve even greater survival benefit.

Objective The aim of this study is to investigate the expression of Survivin in laryngeal car-cinoma and to elucidate the relationship between cell proliferation and the expression of Survivin .Methods The expression and DNA quantification of Survivin in 63 cases of laryngeal carcinoma and 15 cases of normal laryngeal tissues were detected by immunohistochemical staining ( SP) and flow cytometry .Results Forty-five sections of laryngeal carcinoma were positive for survivin as determined by immunohistochemical staining ,but Survivin was undetected in normal laryngeal tissues .The intensity of Survivin expression was significantly increased with tumor differentiation and lymph node metastasis (P<0.05),and was negatively correlated with age ,gender and clinical stage of patients(P>0.05).High DNA index(DI)and proliferation index(PI)were detected in laryngeal carcino-ma(P<0.05),and PI was positively correlated with expression of Survivin (P<0.05).Conclusion Survivin overexpression triggers laryngeal carcinoma cell hyperproliferation ,especially in development of laryngeal carcino-ma.These data identify Survivin as an important target in laryngeal carcinoma and provide a translational pathway for developing new therapies against this target .

BACKGROUND: Salvia miltiorrhiza is widely used to treat angina cordis, ischemic stroke and other ischemic cardiovascular diseases. However, the effects of Salvia miltiorrhiza on ovariectomized rats remain unclear.OBJECTIVE: To observe the effects of Salvia miltiorrhiza on the body mass, food intake, and levels of blood lipids and malondialdehyde (MDA) in ovariectomized rats.DESIGN: A completely randomized and controlled experiment.SETTING: Institute of Physiology and Psychology, School of Basic Medical Sciences, Lanzhou University.MATERIALS: The experiment was performed in the Key Laboratory of Pre-clinical Study for New Drugs of Gansu Province and the laboratory of Institute of Physiology and Psychology, School of Basic Medical Sciences, Lanzhou University from November 2005 to December 2006. Twenty-four healthy female SD rats of 3 months old and (220±2) g were selected. Salvia miltiorrhiza water decoction (equal to 1 g/mL crude drug) was identified and extracted by Drug Control Institute of Gansu Province; MDA kit was purchased from Nanjing Jiancheng Institute of Bioengineering.METHODS: ①The rats were randomly divided into three groups with 8 rats in each group: sham-operated group,ovariectomized group and Salvia miltiorrhiza group. The rats were underwent a bilateral ovariectomy except those in the sham-operated group, which were subjected to a removal of bilateral fat as much as ovariectomized group with the ovaries remained. Rats in sham-operated group and ovariectomized group freely drank water; rats in Salvia miltiorrhiza group freely took 1% water extracts from Salvia miltiorrhiza postoperatively, and the concentration of Salvia miltiorrhiza gradually increased to 12% on the eighth day, which was lasted until the end of the experiment (55 days). ②The food intake of rats in each group was monitored daily, and the body mass was measured every five days. At the end of the experiment, femoral artery blood samples of rats were collected to determine the levels of blood lipids. At the same time,MDA was measured according to the kit.MAIN OUTCOME MEASURES: The body mass, food intake, levels of blood lipids and malondialdehyde in each group.RESULTS: Twenty-four rats all entered the result analysis. ①The body mass of rats in 3 groups was nearly the same before operation (P ＞ 0.05). While the body mass in ovariectomized group on the postoperatively 10th, 20th, 25th, and 55th days was significantly higher than those in sham-operated group (P ＜ 0.01). The body mass in Salvia miltiorrhiza group on the postoperatively 20th, 25th, and 55th days was significantly lower than those in ovariectomized group (P ＜0.05-0.01). ②The food intake in ovariectomized group on the postoperatively 15th, 40th, and 55th days was significantly higher than those in sham-operated group (P ＜ 0.05-0.01), and that in Salvia miltiorrhiza group was significantly lower than those in ovariectomized group at those 3 time points (P ＜ 0.05-0.01). ③At the end of the experiment, the levels of total cholesterol, low density lipoprotein cholesterol, high density lipoprotein cholesterol and triglyceride in ovariectomized group were significantly higher than those in sham-operated group (P ＜ 0.05-0.01). The levels of triglyceride and MDA in Salvia miltiorrhiza group were significantly lower than those in ovariectomized group (P＜0.01, 0.05).CONCLUSION: Salvia miltiorrhiza can significantly reduce the body mass and levels of triglyceride and MDA in ovariectomized rats.

Purpose　Both the working standard and corresponding sample(GRAN75)were first calibrated by WHO international standard for G-CSF.Methods　MTT method by NFS-60 cells was used. The results were calculated by (4,4)method.Results　Three batchs working standards were prepared,two batchs were freeze-dry and the prescription was same as WHO international standard for G-CSF,one batch was injection and the prescription was same as corresponding sample(GRAN75).The biological potency and the FL% of average potency were 3.062×106 、4.276×106IU/ampoule、1.635×107IU/ml and 5.529%、4.291%、4.244% for working standard, and 1.880×107IU/ml and 5.175% for corresponding sample,respectively.Conclusion　The working standard which calibrated could be used as working standard in the measurement of rhG-CSF biological activity.

Purpose　The determination method of rhG-CSF bio logical activity by NFS-60 cells was studied in accordance with WHO internation al standard for G-CSF.Methods　MTT method was adopted.R esults　The chomosone number of NFS-60 cell was 39, about 1×105 cells/ml to 1×107 cells/ml,the dying color showed gradient when the NFS-60 c ells was dyed by MTT,the method(4,4) was adopted in the determination of rhG-CS F biological activity,the average FL% of potency was 13.560% for single exprimen t,and the CV of inter-assay and intra-assay was lower than 10% and 10.109%,respectively.Conclusion　method(4,4) can be us ed in the determination of rhG-CSF biological activity,and the results can guid e the study and the manufacture of rhG-CSF effectively.

Objective To study the expressions of S100A4 and urokinase plasminogen activator(uPA) in pancreatic cancer cells and their correlation with patients prognosis.Methods The expressions of S100A4 and uPA were examined in 63 surgical specimens of primary pancreatic carcinoma by suing immunohistochemistry PV methods,and correlation of their expressions and prognosis of pancreatic cancer was analyzed.Results ( 1 ) Positive immunostaining for S100A4 and uPA was observed in 74.6％ (47 cases) and 65.1％ (44 cases) of 63 pancreatic cancer samples respectively.(2) The positive expressions of S100A4 and uPA were significantly correlated in pancreatic cancer( P =0.000,r =0.567 ).( 3 ) The expression of S100A4 significantly correlated with TNM stages( P =0.002 ),lymph node metastasis ( P =0.002 ) and distant metastasis ( P =0.007 ).The expression of uPA had a significant correlation with TNM stages ( P =0.002),lymph node metastasis ( P =0.001 )and differentiation(P =0.003).(4) Kaplan-Meier survival analysis showed that the median survival (21 months) of patients with S100A4 ( - ) was significantly longer than the median survival ( 9 months) of the patients with S100A4( + )(P=0.000) ;the median survival(9 months) of patients with uPA( + ) was significantly shorter than the median survival ( 18 months) of the patients with uPA ( - ) ( P =0.000) ; the median survival(23 months)of 13 patients with S100A4( - )/uPA( - )was significantly longer than the median survival of other cases ( Log-rank test,x2 =54.444,P =0.000).( 5 ) Cox regression model ( x2 =53.974,P =0.000 )analysis suggested:the differentiation(P =0.004),lymph node metastasis(P =0.017) 、S100A4( + ) expression ( P =0.000) and uPA ( + ) expression ( P =0.001 ) were independent prognostic factors for pancreatic cancer.Conclusion S100A4 and uPA are highly expressed in pancreatic cancer cells,and S100A4 expression has positive correlation with uPA expression,which indicates that the overexpression of S100A4 and uPA maybe poor prognosis factors for pancreatic cancer patients.S100A4 maybe promote extracellular matrix and basement membrane degradation by up-regulation of uPA protein expression,and ultimately promote tumor invasion and metastasis,which is not favorable to prognosis.They may be potential indicators of metastasis and predictors for prognosis of pancreatic cancer.

Objective To investigate the relationship between plasma homocysteine (Hcy), Kv1.3 channel and cardiac troponinI (cTnI) in patients with acute ST-segment elevation myocardial infarction (STEMI). Methods According to the level of Hcy, 80 STEMI patients were divided into STEMI with Hhcy group (Hcy > 15 μmol/L, n=41) and control group (STEMI group, Hcy≤15μmol/L, n=39). The Hcy, blood lipid and cTnI were detected with automatic biochemistry analyzer, respectively. Peripheral lymphocytes were isolated by ficoll density gradient centrifugation. Real-time PCR was used to detect mRNA expression of Kv1.3, and Western blot assay was used to detect protein expression of Kv1.3. Results cTnI concentrations were obviously higher in STEMI with Hhcy group than those in STEMI group (μg/L:22.997 ± 5.880 vs. 12.881 ± 6.343;P<0.01). Multiple linear regression analysis showed that age, gender, hypertension, diabetes mellitus, smoking, family history, total cholesterol (TC), triglyceride (TG), high density lipoprotein-cholesterol (HDL-C) and low density lipoprotein-cholesterol (LDL-C) had no obvious influence on Hcy (P>0.05). The relative expression levels of Kv1.3 mRNA and protein were significantly higher in STEMI with Hhcy group (1.35±0.14, 0.85±0.12) than those in STEMI group (1.00 ± 0.07, 0.64 ± 0.05, P<0.05). Moreover, there was a positive relation between Hcy level and the mRNA and proteinexpression of Kv1.3 channel (r=0.299, r=0.542, P<0.05). There was a positive relation between protein expression levels of Kv1.3 channel and cTnI (r=0.644, P<0.05). Conclusion Our results support that Hcy could exacerbate the concentration of cTnI through playing an important role in the Kv1.3 mRNA and protein expression in lymphocytes.

Objective To investigate the effects of 1,25-dihydroxyvitamin D3[1,25 (OH)2D3] on cell proliferation in hu?man glomerular mesangial cells and it′s effects on the regulation of mTOR/p70s6K signaling pathway in this cell line. Meth?ods The cultured human mesangial cells at passage 3-7 were divided into four groups:control group,VD group (addition of 10-8 mol/L of 1,25-dihydroxyvitamin D3 ),R group (addition of 5 mg/L of rapamycin) and R+VD group(addition of 5 mg/L ra?pamycin combined with 10-8 mol/L of 1,25-dihydroxyvitamin D3). Drug incubation last 48 h. The effect of mesangial cell pro?liferation was measured by CCK-8 colorimetric assay. The cell cycles were measured by flow cytometry. The expression of mTOR and p70s6K were detected by immunofluorescence. Results (1) The absorbance of A450 was higher in control group than that in VD group than that in R group than that in R+VD group. But the inhibition rate (IR) was lower in control group than that in VD group than that in R group than that in R+VD group. All comparisons were of statistic significance. ( 2) Cells in G1 phase were higher while cells in G2/M and S phases as well as proliferation rate (PI) were lower in control group than those in VD group than those in R group than those in R+VD group. All comparisons were of statistic significance except in?dexes between group R and group VD. (3) mTOR and p30s6K expressions in mesangial cells were higher in control group than those in VD group than those in R group than those in R+VD group. All comparisons were of statistic significance ex?cept indexes between group R and group VD. Conclusion 1,25-dihydroxyvitamin D3 might inhibit mesangial cell prolifera?tion significantly through mTOR/p70s6K signaling pathways.

OBJECTIVE To study the changes in inflammatory factors caused by prostatitis.METHODS SD rats were castrated under sterile conditions.E2 0.25 mg· kg-1+ T 0.25,0.5 and 1.0 mg· kg-1 were given sc for 30 d,respectively.Serum samples were taken and levels of E2,T and dihydrotestosterone (DHT) were detected by ELISA.Pathological changes of prostate tissue were observed by HE staining.The expressions of tumor necrosis factor-alpha (TNF-α),cyclooxygenase-2 (COX-2) and macrophage inflammatory protein 1alpha (MIP-1α) in prostate were detected by immunohistochemistry.RESLULTS ELISA detection showed that E2 levels were significantly increased [(80±7) ng· L-1,P＜0.01] in E2 0.25 mg· kg-1 group and that T levels were significantly decreased [111 ±6 vs (111 ±5) nmol· L-1,P＜0.05]in E20.25 mg ·kg-1 and E2+T 0.25 mg·kg-1 groups compared with the sham-operated group.E2 was significantly increased [(80±7) ng· L-1,P＜0.01] in E20.25 mg· kg-1 groups compared with the castrated control.The sham and castrated control showed normal glandular epithelium without leukocyte infiltration.In E2 0.25 mg·kg-1 group,extensive infiltration of inflammatory cells was found in the glandular lumens,suggesting the occurrence of chronic prostatitis.In each E2+T groups,fewer inflammatory cells were noted in the stroma around glands.The expressions of TNF-m COX-2 and MIP-1α in sham group were negative or low,while those of castrated control and E2 0.25 mg· kg-1 groups were high,especially in E2 0.25 mg· kg-1 group.The expressions of TNF-α,COX-2 and MIP-1α in each E2+T group were significantly decreased.CONCLUSION Testosterone can inhibit prostatitis and the expression of inflammatory factors,such as TNF-α,COX-2 and MIP-1 α,in castrated SD rats initiated by estrogen.