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 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 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.

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.

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.

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.

A child case of acute myocardial ischemia induced by symptomatic myocardial bridge (SMB)was reported and relevant literatures were reviewed in this article.The case and literatures indicate that myocardial bridge is a rare congenital coronary artery anomaly in children.The pathogenesis may be related to stenosis,spasm,arteriosclerosis of coronary artery because of myocardial bridge compression during systole.Clinical manifestations include chest pain,malignant arrhythmia,syncope,cardiac arrest and myocardial infarction.The diagnosis of myocardial bridge is based on cardiac imaging.SMB patients should be first treated with β-adrenergic; if ineffective,surgical treatment or implantable automated cardiac defibrillator shonld be chosen.The patients complicated with hypertrophic cardiomyopathy have worse conditions with high mortality.

Objective To study the effect of recombinant human endostatin (rhES)on the radiosensitivity of esophageal squamous cells KYSE-150 and its preliminary mechanism.Methods Cells were divided into four groups:control group without treatment,rhES group treated with recombinant human endostatin,radiation alone group exposed with X-rays,and combination group exposed with X-rays plus endostatin.Colony formation assay was used to measure cell survival fraction.A single-hit multi-target model was used to fit cell survival curve and calculate the sensitive enhancement ratio (SER).Influence of rhES combined with X-ray radiation on cell cycle and apoptosis was measured by flow cytometry.Expressions of Cyclin B1,Cyclin D1,Bcl-2 and Bax mRNAs were determined by RT-PCR.Protein expressions of HIF-1α,VEGF,and VEGFR were determined by Western blot.Results D0,Dq and SF2 value of KYSE-150 cells decreased along with the concentration of rhES.At D0dose,the SER for 100 and 200 μg/ml rhES was 1.14 and 1.27,respectively.Compared with the radiation alone group,the apoptosis rate and bax expression increased,while the expressions of VEGF and HIF-1α decreased in the combination group (t =7.97,3.02,117.55,7.22,P ＜ 0.05).Conclusions rhES has radiosensitive effect on esophageal carcinoma cells KYSE-150 in vitro by inhibiting the expressions of HIF-1α and VEGF,regulating bax expression,and inducing apoptosis.

Objective To prospectively study the dynamic variation of vascular endothelial growth factor (VEGF),the short-term efficiency and the tolerability of the esophageal cancer patients treated by radiotherapy combined with thalidomide.Methods The serum samples of 86 esophageal cancer patients were collected before,during and after the radiotherapy.The VEGF levels were assayed by enzyme-linked immunosorbent assay (ELISA).3 patients interrupted the treatment because of intolerance radiotherapy.Based on the changes of VEGF level,32 esophageal cancer cases whose VEGF levels increased or remained were assigned to the group to which thalidomide was given during the whole course of radiotherapy.The rest 51 patients whose VEGF level decreased received radiotherapy without thalidomide during the whole course.In addition,30 healthy cases were included in control group.Then the efficiency and safety of the introduction of thalidomide in radiotherapy were investigated. Results The VEGF levels of 86 esophageal cancer cases were significantly higher than the 30 healthy control cases ( t =5.07,P ＜ 0.01 ),which were also correlated with the primary tumor size ( t =4.55,P ＜ 0.01 ),lymph node metastasis ( t =7.50,P ＜0.01 ),histological type( F =3.40,P ＜ 0.01 ) and clinical stage ( t =2.52,P ＜ 0.0 l ).However,it was irrelevant to the lesion site,distant metastasis,X-ray pathologic type,gender or age ( P ＞ 0.05).For the thalidomide involved group, the VEGF level after radiotherapy was significantly lower than during radiotherapy (t =2.37,P ＜0.05 ) with an effective rate of 71.88％.For the rest 51 cases without using thalidomide,the effective rate was 78.43％ yet there was no significant difference between the VEGF levels during and after radiotherapy ( t =0.18,P ＞ 0.05 ).62.50％ patients reported symptoms of dizzy and burnout after using thalidomide,while this incidence was 15.69％ for the rest patients (x2 =19.28,P =0.000).For the groups with or without thalidomide combination,the sleepiness incidences were 18.75％and 1.96％,respectively (x2 =5.168,P =0.023 ); the Ⅲ - Ⅳ grade esophagitis incidences were 12.50％ and 11.76％,respectively (x2 =0.061,P =0.806) ; the Ⅱ - Ⅳ grade leukocyte decrease incidences were 6.25％ and 9.80％,respectively (x2 =0.026,P =0.872) ; the Ⅲ - Ⅳ grade platelet descend incidences were 3.13％ and 5.88％,respectively (x2 =0.002,P =0.965 ) ; the Ⅲ - Ⅳ grade nausea and vomiting incidences were 9.38％ and 27.45％,respectively (x2 =2.913,P =0.088 ). No anaphylaxis was observed. Conclusions Thalidomide can decrease the VEGF expression level of esophageal cancer patients.Patients treated with thalidomide show good tolerance and compliance.