To explore whether aberrant methylation in the promoter of p16 gene was associated with development and clinicopathological characteristics of colorectal cancer. Methylation-specific PCR(MSP) was used to detect hypermethylation of p16 gene in tumor tissues obtained from 32 patients with colorectal cancer. The results showed that the hypermethylation of p16 promoter was detected in 40.6% of tumor tissues. p16 hypermethylation in patients with Dukes stages of C and D tumors (63%) was higher than that in the stages of A and B tumors (25%). The highly and intermediately differentiated carcinomas had lower positive rate (30%) than the poorly differentiated carcinoma. Furthermore, the hypermethylation of p16 gene in tumor tissue from patients with the lymph node metastasis was different from that without lymph node metastasis (P

Objective To explore whether hypermethylation in the promoter of p16 gene and protein of p16 were associated with development and clinicopathological characteristics of colorectal cancer. Methods Methylation-specific PCR ( MSP) and immunohistochemistry SP were used to detected hypermethylation of p16 gene and p16 protein in tumor tissues from 32 patients with colorectal cancer. Results The hypermethylation of p16 gene was detected in 40. 6% of tumor tissues. The protein of p16 promoter was detected in 75% of tumor tissues. The hypermethylation of p16 gene was detected in 63% in Dukes stages of C and D tumors. The protein of p16 promoter was detected in 69% of tumor tissues. The hypermethylation of p16 gene was detected in 25% in the stages of A and B tumors. The protein of p16 promoter was detected in 81% of tumor tissues. The hpermethylation of p16 gene was detected in 100% in low differentiated carcinomas. The protein of p16 promoter was detected in 20% , the hypermethylation of p16 gene was detected in 30% , in the high and mediate differentiated carcinomas, the protein of p16 promoter was detected in 85%. Furthermore, the hypermethylation of p16 gene was detected in 63% in the lymph node metastasis and 25% in without lymph node metastasis. The protein of p16 promoter was detected in 65% in rectum and 100% in colon. Conclusions Our study demonstrated that p16 hypermethylation and protein were associated with the development of colorectal cancer and could be used as a putative prognostic indicator for this malignancy.

Objective To explore the effects of low load exercise on the gait and balance in patients with Parkinson's disease. Methods 12 inpatients with Parkinson's disease from May to August, 2015 accepted low load exercise on Power Rehabilitation System 14 times in 2 weeks, with the medication as before. They were assessed with 3D gait analysis and Berg Balance Scale before and after treatment. Results The step length, stride length and walking speed improved after treatment (P<0.05), and improved more in the seriously affected limbs. There was no difference in cadence, stride time, step time, single support, double support, opposite foot off and opposite foot contact after treatment (P>0.05). The scores of Berg Balance Scale improved after treatment (P<0.05). Conclusion The low load exercise can improve the gait and balance in patients with Parkinson's disease.

Objective To evaluate the influence of fast-track surgery in perioperative period on the clinical outcomes of patients at nutritional risk in respectable esophageal cancer surgery perioperatively.Methods A total of 170 esophageal carcinoma patients receiving radical operation in our hospital from January 2008 to December 2013 were randomly divided into two groups by simple random method (n =85 each):one group was treated with the new concept of FTS-based on nutritional risk screening (FTS group),and the other control group received conventional perioperative management (CPM group).The postoperative first passage of flatus and defecation,time to drainage tube removal,postoperative hospital stay,and morbidity of the postoperative complication were recorded and compared.Results The time to drainage tube removal and length of postoperative hospital stay were significantly lower in the FTS group than those in the CPM group,and the overall postoperative complication rate was 7.06％ (6/85) in the FTS group and 20.00％ (17/85) in the CPM group (all P ＜0.05).In FTS group,the first flatus time was (59.01 ±2.73) h,the first defecation time was (3.35 ± 1.37) d,removing time of chest tube was (2.76 ±0.34) d,and postoperative hospital days was (8.16 ± 0.80) d; in the control group,they were (90.16 ±2.82) h,(4.78 ± 1.74) d,(4.39 ±0.25) d,and (10.93 ± 1.39) d respectively,showing significant differences (all P ＜0.05).The operative time was similar between these two groups.Conclusion The new concept of FTS by nutrition risk screening and intervention apparently can accelerate recovery after esophagngastrectomy,reduce the rate of overall complications,promote bowel function recovery,and decrease morbidity in the perioperative period for patients with esophageal carcinoma.

Objective To investigate the effect of different doses of ketamine on inflammatory cytokines in rabbits with severe burn at early stage and preliminarily approach its regulatory action on early stage of inflammatory reaction due to stress of trauma.Methods Forty healthy male New Zealand rabbits were randomly divided into four groups in accord with the random number table method: normal control group, scald model group, ketamine analgesia group and ketamine anesthesia group. Before scald, pentobarbital sodium was used for anesthesia, afterwards catheters were inserted into internal jugular vein and internal carotid artery respectively ready for use, and 24 hours later, Ⅲ degree scald at the animal back and buttocks occupying 30% total body surface area (TBSA) was performed as the scald model for all the rabbits except those in normal control group. In ketamine analgesia group, after scald for 0.5 hour, 0.5 mg/kg ketamine intravenous injection was given to the rabbits as the loading dosage and then persistent intravenous pump infusion of 9μg·kg-1·min-1 ketamine was applied for all together 24 hours. In ketamine anesthesia group, after scald for 0.5 hour, 1.5 mg/kg ketamine intravenous injection was given to the rabbits, and then persistent intravenous pump infusion of 45μg·kg-1·min-1 ketamine was applied for 4 hours to maintain systemic anesthesia. In normal control and scald model groups, only intravenous infusion of equal amount of normal saline was given to the rabbits. The amount of intravenous transfusion in each group and the total dosages of ketamine used in ketamine analgesia group and ketamine anesthesia group were recorded. Before scald and 0.5, 6, 12, 24 hours after scald, arterial blood gas analyses were made, and the levels of serum interleukins (IL-1, IL-6) and tumor necrosis factor-α (TNF-α) were determined.Results Although the indexes of blood gas analysis were changed in the four groups, they were all in the normal range, showing that the respiratory function was in the normal range and indirectly reflecting that the circulatory function was also in the normal range, thus the effects on cytokines by factors of respiratory and circulatory functions were ruled out. The levels of IL-1, IL-6 and TNF-α before scald showed no statistically significant differencesamong the four groups (allP > 0.05). From 0.5 hour after scald, the levels of IL-1, IL-6 and TNF-α were markedly higher in model group than those of normal control group [IL-1 (ng/L): 30.27±0.93 vs. 13.79±1.11, IL-6 (ng/L): 47.22±1.49 vs. 46.31±4.12, TNF-α (ng/L): 243.39±20.85 vs. 190.95±14.97, allP < 0.05], and the situation continued until 24 hours after scald; the levels of IL-1, IL-6 and TNF-α from 6 hours after scald were significantly decreased in ketamine analgesia and ketamine anesthesia groups compared with those in the model group, and from 12 hours after scald, the degrees of descent in levels of the above indexes in ketamine analgesia group were more obvious than those in ketamine anesthesia group [IL-1 (ng/L): 19.28±2.51 vs. 40.12±10.31, IL-6 (ng/L): 52.10±4.23 vs. 72.20±10.11, TNF-α (ng/L): 246.03±20.74 vs. 313.71±27.34, allP < 0.05].Conclusion The low-dose ketamine analgesia and ketamine anesthesia have certain degree of inhibitory effect on the expression and release of inflammatory cytokines at the early stage in rabbits with severe burn, the effect of long-term low-dose ketamine analgesia being more significant.

To obtain and analyze the sequence of the nucleocapsid gene from bovine coronavirus, and to produce the fusion protein of the N gene in E.coli in order to use this recombinant protein for the study of bovine coronavirus. The N gene of BCV-DQ strain was amplified by RT-PCR, in which the primers were designed on the basis of N gene sequence of BCV-Mebus strain. The PCR products of 1 347 bp in length were cloned and sequenced, and then inserted into the prokaryotic vector pET30a. The recombinant plasmids were then transformed into Escherichia coli BL21 and identified by SDS-PAGE and Western blot assay. ELISA assay was optimized of N protein as the coating antigen to detect the viruses in the clinical samples. In comparison with 6 BCV strains in GenBank, the sequence identity was proved to be more than 98.3%. Result in SDS-PAGE showed that the fusion protein had a molecular weight of 60 ku, and could be specifically recognized by mouse serum against BCV. The indirect ELISA was used to test 256 serum samples collected from Heilongjiang province and 65.23% samples were positive. On testing field samples, an overall agreement of 95.31% was generated between the the neutralization test of viruses (VN) and indirect ELISA. It is apparent that the N gene was highly conservative and is expressed in E. coli in high level,also the prokaryotic expression products of this gene show a fine reactiongenicity in immune responses. It was also suggested that the N protein may be a useful antigen for sero-diagnosis and epidemiological investigation of BCV.

Objective In a prospective randomized controlled pilot study, effects of postoperative pulmonary complications on a conservative treatment surgery (CTS) and fast track surgery (FTS) treatment regimen in non-small cell lung cancer (NSCLC) patients undergoing pulmonary lobectomy were compared.Methods Eighty patients who underwent radical pulmonary lobectomy surgical treatment for non-small cell lung cancer disease from January 2008 to May 2010 in our hospital were random assigned to either fast track surgery treatment (40 FTS group) or conservative treatment surgery regimen (40 CTS group). Study endpoints were pulmonary complications ( pneumonia, atelectasis, prolonged air leak ＞ 7 days); Further parameters assessed in the postoperative course of patients were the need for postoperative mechanical ventilation, temperature at the end of the operation, length of stay (LOS) on intensive care unit (ICU) and day of discharge. Results The rate of postoperative pulmonary complications was 34. 21% in CTS group and 8. 33% in FTS group ( P ＜0. 05). Median length of stay on ICU was comparable in both groups ( 1 day),but the day of discharge was significantly different in both groups [( 11. 1 ±3.6)d vs ( 16. 6 ±5.7)d, P ＜0. 01]. Conclusion Using this fast track clinical pathway, the rate of pulmonary complications could be significantly decreased as compared to a conservative treatment regimen. Our results supported the implementation of an optimized perioperative treatment in lung surgery for non-small cell lung cancer patients undergoing radical pulmonary in order to reduce pulmonary complications after major lung surgery.

Aim: To develop a practical synthetic route of raltegravir, a drug for HIV treatment. Methods: Raltegravir was synthesized through an eight-step process including aminonitrile formation, protection with benzyloxy-carbonyl group, conversion of the nitrile to the amidoxime, cyclization to form hydroxypyrimidinone, N-methyla-tion, amidation with microwave-assistance, deprotection, amidation with acyl chloride. Results: The overall yield of the eight-step synthesis is about 12. 0% and the structure of the target compound was confirmed by ~1H NMR, ~(13)C NMR, LR-MS and HR-MS. Conclusion: The reported synthetic process of raltegravir highlights the advantages in terms of readily available starting materials, convenient operation and low cost.

Objeetive To investigate the clinical efficacy of treating stomach cance belonging to spleen and stomach deficient cold type with self-designed formula and chemotherapy.Methods 100 patients of stomach cancer belonging to stomach and spleen deficient cold were randomly grouped into a control group and a treatment group,with 50 cases in each group.The control group was only treated with chemotherapy,and the treatment group was additionally treated with self-designed Chinese formula on the basis of the control group.After 3 months' treatment,the curative effect,the improvement of life quality,the changes of body weight,the improvement of peripheral blood and immune function,and the occurrence of adverse reaction were observed.The survival rate of patients after 1,3 and 5 years' treatment was investigated.Results ① Short term curative effect:the total effectiveness of the control group and the treatment group was 60.0％ (30/50) and 30.0％ (15/50) respectively,with the treatment group being better than the control group(Z=-2.100,P＜0.05);② life quality:theimprovement rate of life quality in the treatment group and the control group was 64.0％ (32/50) and 20.0％ (10/50) respectively,with the treatment group being better than the control group (Z=-3.259,P＜0.05); ③ body weight:the improvement rate of body weight in the treatment group and the control group was 24.0％ and 16.0％ respectively,and there was no significant difference (Z=-1.815,P＞0.05) ; ④ peripheral blood:the peripheral blood abnormality rates of treatment group was obviously lower than the control group (Z=-4.286,P＜0.05) ;⑤ immune function changes:CD4+、CD8+、and the activity of NK cell of two groups after the treatment [the treatment group was (40.8 ± 4.1)％,(26.1-3.2)％,(83.57-2.52)％ respectively; the control group was (31.7±2.9)％、(33.8±3.0)％、(92.88±3.83)％ respectively] wasobviously improved compared with the same group before the treatment [the treatment group was (34.2±3.5)％,(31.0±2.0)％,(85.01±2.92)％ respectively; the control group was (34.0±3.2)％,(30.9±2.2)％,(85.02± 3.48) ％ respectively].The immune indexes (CD4+、CD8++、CD4+/CDg+and the activity of NK cell)in the treatment group presented significant difference (P＜0.01) than the control group after the treatment.⑥adverse reaction:compared with the control group,the occurrence rate of adverse reaction of the treatment group was obviously lower than the control group,and there was significant difference (Z=-5.297,P＜0.05).⑦survival rate:the survival rate of patients after 1,3 and 5 years' treatment in the treatment group was better than the control group,and there was significant difference (x2=5.263,12.96,7.895,P＜0.05).Conclusion The combination of self-designed Chinese formula and chemotherapy could effectively relieve the clinical symptoms,improve the immunity,reduce the occurrence of adverse reaction,and improve the life quality of patients with stomach cancer belonging to stomach and spleen deficient cold type.

OBJECTIVE To study the radiose-nsitization by Topotecan on human nasopharyngeal carcinoma in nude mice. METHODS ①To study the maximum tolerance dose of TPT and detect the effective rate of TPT and RT on nude mice. ② Plan of radiosensitization practice:53 nude mice xenografts were distributed to 5 groups:RT 20 Gy group,RT 40 Gy group,TPT 12.5 mg/kg group,TPT 12.5 mg/kg+RT 20 Gy group and the controlgroup. After treatment,the volume of tumors were measured every 3 days in order to value the effective rate [complete remission(CR) + partial remission(PR) ]and regrowth delay time(TGD) and to fit the growth curve. RESULTS This study showed that the effective rates had significant difference among RT20 Gy+TPT 12.5 mg/kg group,RT20 Gy group and TPT12.5 mg/kg group,while that of RT20 Gy +TPT 12.5 mg/kg group and RT40 Gy group had no statistical difference. SER reached to 1.34. CONCLUSION Topotecan has been shown a radiosensitizing effect on human nasopharyngeal carcinoma in vivo.