The chronic-hypoxia-resistant gastric cancer cell line was established, and its biological characteristics were explored and compared with the parental cell line. Gastric cancer cell lines were cultured under the degressive oxygen concentration. Cell doubling time was calculated by cell counting method. Chemo-resistance ability of cells was tested by MTT assay. Irradiation tolerance of cells was evaluated by colony forming method. Cell cycle distribution was tested with flow cytometry. Invasive ability was tested by Transwell method. The expression levels of GLUT-1 and HIF-1α were detected by using Western blot. MNK45/HYP cells successfully survived under the 1% concentration of oxygen and its cell doubling time was 35.01±1.02 h, while that of MNK45 was 27.35±0.83 h (P<0.01). The percentage of MNK45/HYP cells in G(0)/G(1) stage was (58.3±6.1)%, and that of MNK45 cells was (42.2±6.0)% (P<0.05). Comparing with the parental cells MNK45, drug resistance indexes of 5-Fu, PTX, OXA, Sn38, GEM and VP16 in MNK45/HYP cells were respectively 5.3, 1.3, 3.6, 2.2, 4.8 and 4.4. Colony forming ability of MNK45/HYP cells after irradiation was also significantly higher than MNK45 cells. The invasive number of MNK45/HYP cells was 107.7±17.5, while that of MNK45 cells was 59.0±9.9. The expression levels of GLUT-1 and HIF-1α in MNK45/HYP cells were significantly higher than those in MNK45 cells. MNK45/HYP cells hold biological characteristics of hypoxia tumor with good tolerance to chronic hypoxia, and can be used for the research of solid tumor under chronic hypoxia condition.

In order to investigate the effects of short hairpin RNA (shRNA) on the expression of Survivin, cell cycle and cell proliferation in MCF-7 cells, using a pEGFP vector which contained a U6 promoter shRNA plasmid targeted against survivin was constructed and transfected into MCF-7 cells. The change of the expression of Survivin and cell proliferation rates were detected by semiquantitative reverse transcription polymerase chain reaction (RT-PCR) and MTT methods respectively. The change of cell cycle after transfection was analyzed by flow cytometry. The results indicated that the recombinant plasmid containing Survivin shRNA was constructed successfully, which could suppress the expression of Survivin at mRNA and protein level. The growth of MCF-7 cells was arrested in G1 phase of the cell cycle and the proliferation activity was suppressed after transfection. It was concluded that Survivin shRNA plasmid could knock down the expression of Survivin in MCF-7 cells specifically. In addition, Survivin shRNA plasmid could lead to G1 arrest and inhibit the proliferation of MCF-7 cells, which suggested that Survivin shRNA might be used as a new therapeutic method for breast cancer.

Objective To investigate the efficacy of skin scraping plus movable cupping in treating neck-shoulder syndrome. Method One hundred and eighty patients with neck-shoulder syndrome were randomly allocated to treatment and control groups. The treatment group received skin scraping plus movable cupping and the control group, acupuncture. Result The total efficacy rate was 88.8% in the treatment group and 77.5% in the control group; there was a statistically significant difference between the two groups (P<0.05). Conclusion Skin scraping plus movable cupping has a good therapeutic effect on neck-shoulder syndrome.

Objective To explore the value of an innovative teaching model which combined problem based learning (PBL) method with case based learning (CBL) method in clinical oncology Teaching.Methods 68 students were divided into the combinational teaching group (30 cases) and the LBL group (38 cases).The combinational teaching group was taught by PBL method combined with CBL method, and this dual track teaching was based on cases and problems.The traditional teaching group was taught by LBL method.The teaching effect was evaluated by students' questionnaire survey and test score.SPSS 13.0 was used to two groups to do t test for statistical analysis in test score and x2 test for degree of satisfaction.Results In the final examination, the score of non-case test of combinational teaching group was similar to that of traditional teaching group (50.30 ± 7.19 vs.52.04 ± 8.01, P=0.358).The combinational teaching group had significant improvement in case analysis test (35.76 ± 5.28 vs.31.80 ± 5.16), and the difference was statistically significant (P=-0.003).In the course of teaching satisfaction survey, the dual track teaching group, compared with the conventional teaching group, has a better effect on self study ability, communication skills, communication skills, and higher satisfaction for teaching, and more willing to continue to carry out teaching (P＜0.05).Conclusion The PBL+CBL combinational teaching model can make a great contribution to improving the teaching quality and satisfaction, and worthy of being popularized and applied.

Objective To observe the clinical efficacy of heat-sensitive moxibustion plus point-toward-point needling in treating poststroke strephenopodia.Method Eighty patients with poststroke strephenopodia were randomized into a treatment group intervened by heat-sensitive moxibustion plus point-toward-point needling and a control group intervened by rehabilitation, 40 cases in each group. In addition to the basic treatment, the treatment group was given heat-sensitive moxibustion plus point-toward-point needling, and the control group was given rehabilitation treatment. Holden's Functional Ambulation Classification (FAC), Fugl-Meyer Assessment (FMA) of the lower-limb motor function, and Tinetti Gait Assessment (TGA) were adopted for evaluation of the two groups, and the clinical efficacies were compared.ResultThe effective rate was 90.0% in the treatment group versus 77.5% in the control group, and the difference was statistically significant (P<0.05); after the treatment,there was a significant difference in comparing the Holden's FAC between the two groups (P<0.05); the FMA score changed significantly after the treatment in both groups (P<0.05), and there was a significant difference in comparing the FMA score between thetwo groups after the treatment (P<0.05); the TGA score changed significantly after the intervention in both groups (P<0.05), and there was a significant difference in comparing the TGA score between the two groups after the intervention (P<0.05).ConclusionHeat-sensitive moxibustion plus point-toward-point needling can produce a significant efficacy in treating poststroke strephenopodia, as it can enhance the effective rate and improve the lower-limb motor function.

The chronic-hypoxia-resistant gastric cancer cell line was established,and its biological characteristics were explored and compared with the parental cell line.Gastric cancer cell lines were cultured under the degressive oxygen concentration.Cell doubling time was calculated by cell counting method.Chemo-resistance ability of cells was tested by MTT assay.Irradiation tolerance of cells was evaluated by colony forming method.Cell cycle distribution was tested with flow cytometry.Invasive ability was tested by Transwell method.The expression levels of GLUT-1 and HIF-1α were detected by using Western blot.MNK45/HYP cells successfully survived under the 1％ concentration of oxygen and its cell doubling time was 35.01±1.02 h,while that of MNK45 was 27.35±0.83 h (P＜0.01).The percentage of MNK45/HYP cells in G0/G1 stage was (58.3±6.1)％,and that of MNK45 cells was (42.2±6.0)％ (P＜0.05).Comparing with the parental cells MNK45,drug resistance indexes of 5-Fu,PTX,OXA,Sn38,GEM and VP16 in MNK45/HYP cells were respectively 5.3,1.3,3.6,2.2,4.8 and 4.4.Colony forming ability of MNK45/HYP cells after irradiation was also significantly higher than MNK45 cells.The invasive number of MNK45/HYP cells was 107.7±17.5,while that of MNK45 cells was 59.0±9.9.The expression levels of GLUT-1 and HIF-1α in MNK45/HYP cells were significantly higher than those in MNK45 cells.MNK45/HYP cells hold biological characteristics of hypoxia tumor with good tolerance to chronic hypoxia,and can be used for the research of solid tumor under chronic hypoxia condition.

In order to investigate the effects of short hairpin RNA (shRNA) on the expression of Survivin, cell cycle and cell proliferation in MCF-7 cells, using a pEGFP vector which contained a U6 promoter shRNA plasmid targeted against survivin was constructed and transfected into MCF-7 cells. The change of the expression of Survivin and cell proliferation rates were detected by semiquantitative reverse transcription polymerase chain reaction (RT-PCR) and MTT methods respectively. The change of cell cycle after transfection was analyzed by flow cytometry. The results indicated that the recombinant plasmid containing Survivin shRNA was constructed successfully, which could suppress the expression of Survivin at mRNA and protein level. The growth of MCF-7 cells was arrested in G1 phase of the cell cycle and the proliferation activity was suppressed after transfection. It was concluded that Survivin shRNA plasmid could knock down the expression of Survivin in MCF-7 cells specifically. In addition, Survivin shRNA plasmid could lead to G1 arrest and inhibit the proliferation of MCF-7 cells, which suggested that Survivin shRNA might be used as a new therapeutic method for breast cancer.

Objective:To explore the effect of laparoscopic surgery on postoperative rehabilitation and stress response of patients with colorectal cancer.Methods:From January 2018 to September 2019, 94 patients with colorectal cancer were selected in the General Hospital of Marine Police of the Chinese People's Armed Police Force.They were divided into observation group (47 cases) and control group (47 cases) according to the random digital table method.The observation group was treated with laparoscopic surgery, while the control group was treated with open surgery.The changes of perioperative indicators, postoperative rehabilitation indicators, postoperative complications and 24-hour stress response indicators were compared between the two groups.Results:The time of lymph node dissection and operation had no statistically significant differences between the two groups (all P>0.05). The amount of bleeding of the observation group [(153.61±27.58)mL] was less than that of the control group [(271.32±23.25)mL] ( t=22.371, P<0.05). In the observation group, the exhaust time [(43.27±6.18)h] and the recovery time of bowel sounds [(20.31±3.28)h] were faster than those in the control group [(67.38±9.91)h and (34.54±5.17)h] ( t=14.153, 15.934, all P<0.05). The incidence of postoperative complications in the observation group (4.26%) was lower than that in the control group (21.28%) (χ 2=6.114, P<0.05). The serum levels of norepinephrine (NE) [(92.13±10.45)μg/L], C-reactive protein (CRP) [(17.38±3.01)mg/L] and Cor [(137.87±23.15)μg/L] in the observation group were lower than those in the control group [(136.27±15.42)μg/L, (26.73±2.34)mg/L and (185.42±27.39)μg/L] ( t=16.245, 16.813, 9.090, all P<0.05). Conclusion:Laparoscopic radical resection of colorectal cancer has good effect on patients with colorectal cancer.It can speed up postoperative rehabilitation and reduce stress response.It is worthy of popularizing.

Objective: Comparison of alkaloid and polysaccharides content in different parts of whole plant of wild aconitum in Nanyang, and to explore the medicinal value of aboveground part and fibrous root of wild aconitum in Nanyang. Methods: The contents of three monoester alkaloids, three diester alkaloids in wild aconitum, radix aconiti carmichaeli, fibrous root and aboveground parts in Nanyang were determined by HPLC method; The content of crude polysaccharides was determined by sulfuric-phenol method, crude polysaccharides content from different tissues of wild aconitum in Nanyang in different harvest time was compared, including wild aconitum in Nanyang, wild radix aconiti carmichaeli in Nanyang, fibrous root, aboveground part. Results: Among the 4 parts of the wild Aconitum in Nanyang, the contents of 3 mono ester alkaloids rang from high to low: wild aconitum in Nanyang＞ wild radix aconiti carmichaeli in Nanyang＞ fibrous root＞ aboveground part (P ＜ 0.05), the contents of three diester alkaloids rang from high to low: wild radix aconiti carmichaeli in Nanyang＞ fibrous root＞ wild aconitum in Nanyang＞ aboveground part (P ＜ 0.05), the order of total alkaloid content from high to low is: wild radix aconiti carmichaeli in Nanyang＞ fibrous root＞ wild aconitum in Nanyang＞ aboveground part (P ＜ 0.05), The content of crudepolysaccharides in different tissues of wild aconitum in Nanyang were in the sequence of wild radix aconiti carmichaeli in Nanyang＞ wild aconitum in Nanyang＞ aboveground part＞ fibrous root (P ＜ 0.05) . Conclusion: In this study, an attempt was made to explore the differences of 6 kind salkaloid content in different parts of whole plant of wild aconitum in Nanyang, which provided scientific basis for the medicinal value of the fibrous roots and the aboveground parts.

OBJECTIVE：To establish HPLC fingerprints of different polar parts of Zhuang medicine Calonyction muricatum ， and to study its spectrum-effect relationship with analgesic and anti-inflammatory effects. METHODS ：The total part ，ethyl acetate part，n-butanol part and water part of C. muricatum were prepared. HPLC fingerprints of different polar parts were established by HPLC method combined with the Similarity Evaluation System of TCM Chromatogramtic Fingerprint （2012A），and the common peaks were identified. Using writhing times and ear swelling degree in mice as analgesic and anti-inflammatory indexes ，analgesic and anti-inflammatory activity of different polar parts of C. muricatum were investigated. The correlation of the common peaks of HPLC fingerprint with analgesic and anti-inflammatory indexes was analyzed by grey correlation analysis ，bivariate correlation analysis and partial least square （PLS） method. RESULTS ： There were 11 common peaks for the different polar parts of C. muricatum ，and 5 components were identified by reference comparison，i.e. neochlorogenic acid （peak 3），chlorogenic acid （peak 5）， cryptochlorogenic acid （peak 6）， isochlorogenic acid A （peak 10），isochlorogenic acid C （peak 11）. The grey correlation analysis showed that the correlation between all common peaks and analgesic and anti- inflammatory effects were greater than 0.6 （except the correlation between peak 6 and analgesic effects ），showing correlation relationship ；the correlation of peaks 3，7 and 10 with analgesic and anti-inflammatory effects were all greater than 0.8，which was highly related. Bivariate correlation analysis showed that the correlation of peak 1，3，4，7，9，10，11 with analgesic and anti-inflammatory effects were all greater than 0.6，showing correlation relationship. PLS method showed that peaks 1，3，4，7，9，10，11 contributed greatly to playing an analgesic and anti-inflammatory role. CONCLUSIONS ：HPLC fingerprints of different polar parts of C. muricatum is established and five common peak components were identified. Neochlorogenic acid ，isochlorogenic acid A ，isochlorogenic acid C and chemical components represented by peaks 1，4，7，9 may be the pharmacodynamic substances of C. muricatum to exert analgesic and anti-inflammatory effects.