According to the yield of intracellular triterpene, the strain GL31 was screened out from 22 strains of Ganoderma collected from various regions. The optimal fermentation conditions were studied by single factor experiments and orthogonal test. The optimal parameters of carbon source, nitrogen source, initial pH, the medium volume in the flask and cell density were obtained. In addition, the metabolic curves of intracellular triterpene and biomass were determined. It was found that the yield of intracellular triterpenes was up to 3.51?10~(-2)g/100 mL medium at the 84th hour. In addition the stram was staticly fermented 144 hours flowing 84hours shaking-flask culture, the IT (intracellular triterpenes) content was increased by 48.6% and the yield of IT was increased by 65% compared with those of the mycelia cultured 84hours using shaking-flask method. This indicated that static fermentation method is helpful for improving the total IT content. The result also show that the highest yield of intracellular triterpenes of mycelia staticly fermented flowing 84hours shaking-flask culture is higher than that of mycelia using shaking flask culture method.

Endostatin is an antitumor molecular targeting angiogenesis of tumor and plays an inhibitory role in tumorigenesis through inhibiting pathological angiogenesis, proliferation and metastasis of tumor. As increasing drugs for targeting therapy aim in clinic, endostatin has become one of hot spots of research on combined treatment of cancer in recent years.

Objective To explore the radiosensitization effect of thalidomide combined with X-ray on esophageal carcinoma TE1 cells.Methods Cell scratch assay Was used to detect the inhibition ability of different concentration of Thalidomide on cell invasion and metastasis.H3-TdR incorporation assay Was used to investigate the inhibition of DNA synthesis in TE1 cells by treated with Thalidomide singly or combination with X-rays.The colony formation assay Was used to analyze the radiosensitization of Thalidomide effect on TE1 cells.Results Thalidomide had obvious inhibition effect on TE1 cell metastasis.DNA synthesis and colony formation,which were correlated with drug concentration.The values D0,Dq and SF2 in TE1 cells were gradually decreased with thalidomide concentration increased.When the concentration of thalidomide was 100μg/ml,the SERD0 and SERDq were(1.4±0.2)and(1.5±0.1),respectively,While the concentration of thalidomide Was 1 50μg/ml,the SERD0 and SERDq were metastasis,DNA synthesis,and significantly enhance the radiosensitizing effect on esophageal carcinoma TE1 cells.

Objective To investigate the radiosensitizing effect of tetrandrine in human esophageal carcinoma cells (TE1) in vitro and its related mechanisms. Methods The cell proliferation was assessed by MTT assay. Colony formation was used to analyze radiosensitivity enhancement by tetrandrine in TE1cells. Western blotting was preformed to measure the cyclin B1 protein levels. Results Tetrandrine inhibited cell growth in a concentration and time depedant manner. The inhibition of proliferation was observed when cells were treated by 1.0, 5.0 and 10. 0 μg/ml tetrandrine for 24 h after irradiation ( P ＜0. 05;F= 3.09, 10.43 and 24. 00, respectively). The inhibition was more significant when cell were treated by 0. 1, 1.0, 5.0 and 10. 0 μg/ml tetrandrine for 48 h than 24 hours after irradiation (F =4. 12,12. 77, 44. 28, and 48.53 respectively ,all P ＜ 0. 01 ). The D0, Dq and SF2 decreased with the increase of the tetrandrine concentration. The maximal sensitizing enhancement ratio was 1.60 with 0. 5 μg/ml tetrandrine. Tetrandrine upregulated the expression of cyclin B1 and removed G2 / M arrest . Conclusions Tetrandrine can enhance radiosensitivity of TE1 cells. This effect may be associated with the increase of cyclin B1 expression to remove G2/M arrest.

Objective To study the radiation sensitive enhancement ratio (SER) of NS398 on esophageal cancer stem cells and adherent tumor cells and analyze the radioresistance related protein expressions.Methods ECA109 esophageal cancer stem cells were cultured in serum-free medium.Expression levels of cell surface maker CD44 + and CD271 + were analyzed by flow cytometry.MTT assay was used to detect cell proliferation after the treatments with NS398 and irradiation(0,4 and 8 Gy).The sensitization effects of NS398 on the parental cells and its spheroid were evaluated by clone formation assay.Western blot assay was performed to determine protein expressions.Results Serum-free medium was successfully applied to isolate the cancer stem cells with spherical properties.CD271 + in the spheroid cells was notable higher than that in the parent cells (t =3.81,P ＜ 0.05).After irradiation,the proliferation rate of parental cells was higher than that in spheroid cells.After the combination treatment of NS398 and irradiation,SF2 value of parental cells was lower than spheroid cells(t =2.91,P ＜ 0.05)and the SER of NS398 on parental cells was greater than spheroid cells.The expressions of Bmi-1,c-Myc,β-catenin and Cyclin D1 in spheroid cells were higher than those in parental cells (t =8.09,7.90,7.50,7.15,P＜0.05).Cyclin D1 expression levels under both cell situations increased after 4 Gy irradiation (t =9.74,6.67,P ＜0.05).Compared to the 4 Gy irradiation alone group,the β-catenin and Cyclin D1 expression levels in both parental cells (t =10.15,12.12,P ＜ 0.05) and spheroid cells (t =3.23,7.45,P ＜ 0.05) decreased in the combination group.Conclusions Esophageal cancer stem cells with high level of CD271 can be isolated with serum-free medium and it is radioresistant where β-catenin and its downstream proteins may be involved.

In recent years, there has been a comprehensive reform of higher medical education in the Medicine School of Wuhan University. According to the need for reform, the teaching of neurology has to be changed from the traditional form to a new form, and be integrated into the clinical pathophysiology and therapeutics (CPPT) courses. Currently neurology in CPPT takes the form of theoretical lectures, case discussions, combined with practical lessons to observe sections under the microscope and clinical practice, for the cultivation of students'!self-learning ability and clinical thinking. In the commissioning process, it exposes some problems in teaching process due to the characteristics of the course in neurology. For example, the knowledge of neuroanatomy is insufficient and review lessons relatively too short, and the teaching effect may be worse due to the fact that teachers have busy clinic work. In addition, students participate in case discussions with less enthusiasm. To solve these problems, we take some measures to promote teaching reform in neurology, such as increasing the review hours of neuroanatomy section in the CPPT neurology, training a group of specialized medical teachers to enrich and stabilize teacher team, adjusting the content and form of discussion class to improve students'!interest and participation, and increasing assistant jobs by the student to assist discussion teaching.

Aim To investigate the effect of astragalo-side IV ( ASIV) on myocardial energy metabolism and mitochondrial biosynthesis in myocardial cells of dia-betic rats induced by streptozotocin ( STZ ) . Methods
50 SD rats at 6 weeks of age were assigned to 5 groups,10 for each group:control group, model group, ASIV 10 mg·kg-1 ·d-1 group, ASIV 20 mg·kg-1 ·d-1 group, ASIV 40 mg·kg-1 ·d-1 group. Except the control group,the remaining 40 were used to estab-lish type 1 diabetes model by the tail vein injection of STZ (35 mg·kg-1 ) . At the end of 16 weeks of treat-ment, left ventricular systolic pressure ( LVSP ) , left ventricular diastolic final pressure ( LVEDP ) and left ventricular maximum rising/falling rate ( ± dp/dtmax ) were tested. Pathological section was observed by HE staining. ATP, ADP, AMP levels were detected by ELISA. The expressions of PGC-1α and NRF-1 protein were assessed by Western blot. The expressions of PGC-1α and NRF-1 mRNA were determined by RT-PCR. Results Compared with control group, model group markedly elevated LVEDP and decreased LVSP, ± dp/dtmax , ATP/AMP and ATP/ADP ratio. Com-pared with model group, low-dose ASIV group did not change significantly,middle-dose ASIV group and high-dose ASIV group obviously decreased LVEDP, and im-proved LVSP, ± dp/dtmax , ATP/ADP and ATP/AMP ratio. Meanwhile, the expressions of PGC-1α and NRF-1 protein and mRNA were increased in a dose-de-pendent manner. Conclusion ASIV could promote mitochondrial biosynthesis, improve energy metabolism in myocardial cells of type 1 diabetic rats by PGC-1αand NRF-1 .

Objective To explore the relationship between pulmonary arterial hypertension (PAH)associat-ed with chronic obstructive pulmonary disease (COPD) and inflammatory reaction(inflammatory cell, bs CRP, IL-8 and TNF-α). Methods According to echocardiography results, the patients (systolic pulmonary artery pressure, SPAP>30 mm Hg) were divided into PAH group(n=36), single COPD group(n=32). All of the patients and 30 healthy subjects (control group) were recruited into the study. Lung function, arterial blood gases, cell differentials in induced sputum, and the levels of serum high sensitivity CRP(hs-CRP), IL-8, TNF-α were determined. Results The incidence of PAH associated with COPD were 53% (36/68),including 27% (3/11) of mild PAH,38% (5/13) of moderate PAH and 64% (28/44) of severe PAH and there were significantly differences in the severity of the spirometric abnormality (χ26.020, P<0.05). The mean PASP and right ventricle wall thickness (RVWT) in PAH group were significantly greater in the patients compared to single COPD[PASP: (52±15)mmHg in PAH group and (23±12) mm Hg in single COPD group, t=3.32,P<0.01 ; PVWT: (5.03±1.04 )mm in PAH group and (3.78±0.57)nun in single COPD group, t=2.36, P<0.05]. The levels of total cell count and neutrophils in induced sputum,hs-CRP,IL-8 and TNF-α in PAH group were higher than those in single COPD group and healthy subjects[toal cell count: (2.84±0.56)×109/L in PAH group and (1.73±0.42)×109/L in single COPD group and (0.68±0.21)×109/L in control group; neutrophils: (2.78±0.52)×109/L in PAH group and (2.57± 0.26)×109/L in single COPD group and (0.63±0.21 )×109/L in control group; hs CRP: (32±12) mg/L in PAH group and (23±11)mg/L in single COPD group and (11±4)mg/L in control group; IL-8: (113±34) ng/L in PAH group and (69±24) ng/L in single COPD group and (38±11) ng/L in control group; TNF-α: (206±63)ng/L in PAH group and (153±54)ng/L in single COPD group and (75±26)ng/L in control group (P<0.05)]. PASP in PAH group was negatively correlated with FEV<,1>% (r=-0.48,P<0.01) and was positively correlated with the levels of serium IL-8 and TNF-α, neutrophils in induced sputum respectively(r=0.43,0.56,0. 47,P<0.01). Conclusion Inflammation reaction is responsible for PAH associated with COPD.

Objective To study the relationship between inflammation and malnutrition in patients with stable chronic obstructive pulmonary disease (COPD).Methods A total of 85 patients with stable COPD and 30 healthy subjects were recruited .All patients were divided into the lower body mass index (BMI,BMI<18.5 kg/m~2) group and normal BMI (BMI=18.5-23.9 kg/m~2) group.Lung function,arterial blood gall,cell differenti-als in induced sputum,and the levels of serum C-reactive protein (CRP),interleukin-8(IL-8),interleukin-6 (IL-6),interleukin-10 (IL-10),and tumor necrosis factor-α(TNF-α) were determined.Results The levels of total cell count and neutrophils in induced sputum were significantly higher in lower BMI group than in normal BMI group and healthy subjects (P<0.05).The forced expiratory volume in 1 second percentage,forced expiratory volume in 1 second/forced vital capacity,and arterial oxygen tension were significantly lower in lower BMI group than in normal BMI group,and the arterial carbon dioxide tension was significantly higher in lower BMI group than in normal BMI group (P<0.05).The levels of serum CRP,IL-8,IL-6,and TNF-α were significantly higher in lower BMI group than those in normal BMI group and healthy subjects (P<0.05).In lower BMI group,BMI was negatively correlated with total cell count (r=-0.492,P=0.0038) and neutrophils (r=-0.501,P=0.0032) in induced sputum and the levels of serum CRP (r=-0.473,P=0.0083),IL-8(r=-0.382,P=0.0421),IL-6(r=-0.422,P=0.0147),and TNF-α(r=-0.416,P=0.0156),respectively.Conclu-sion Local and systemic inflammatory reaction is responsible for malnutrition associated with COPD.

Objective To observe the effects of swimming in cold water on the functioning and structure of the peripheral nerves of diabetic rats,and to compare the effects of seawater and fresh water. Methods Forty SD rats weighing ( 250 ± 20) g were randomly divided into a normal control group (A),a diabetic model group ( B ),a seawater swimming group (C) and a fresh water swimming group (D) with 10 rats in each group.The swimming training was carried on 5 times a week for 8 weeks.At the end of the 4th and 8th week of training,caudal nerve conduction velocity (CNCV) was measured.The nerve structure of the caudal nerves was observed at the end of the 8th week. Results By the 4th week,CNCV had slowed significantly in group B compared with group A,but not in groups C and group D.Compared with group B,CNCV had increased significantly in group C.There was no significant difference in CNCV between groups C and D.At the 8th week,compared with group A,CNCV had slowed in groups B and C.Compared with group B,CNCV was significantly faster in groups C and D.However,there was no significant difference between group C and group D with regard to CNCV.At the end of the 8th week demyelination was observed in the caudal nerves under a light microscope and an electron microscope in groups B,C and D,but the demyelination was milder in groups D and C. Conclusion Swimming in cold water can prevent or delay diabetic neuropathy in diabetic rats.There was no significant difference between seawater and fresh water swimming in terms of its effect on CNCV.