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 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 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 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.

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 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.

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.

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.

ObjectiveTo evaluate the clinical efficacy of bladder hydrodistention and intravesical sodium hyaluronate in the treatment of interstitial cystitis (IC).Methods Twenty-one IC patients received intravesical sodium hyaluronate therapy under combined blockage or intravenous anesthesia.Bladders were perfused with normal saline under 100 cm H2O perfusion pressure and expanded for 10 min,the bladders were then injected through a catheter with 40 mg/50 ml sodium hyaluronate which was released after 1 h.Intravesical perfusion was applied once every week four to six times in a course of treatment.Results The average bladder capacity was extended from 191.6 ± 88.7 ml before expansion to 425.3 ± 79.8 ml after bladder expansion ( P =0.000).The extension was done under anesthesia.There were two suspected bladder ruptures after starting the bladder expansion at 6.5 min and 7.2 min.There was significant gross hematuria in 19 cases,10 min after bladder expansion.After treatment,the catheters were removed 24 h after manipulation in 17 patients; the catheters were removed 72 h after manipulation in two cases with hematuria;the catheters were removed four days after manipulation in the two cases of suspected bladder rupture.Pain was significantly reduced after the catheters were removed and the maximum urinary output increased slightly.The day before the second injection of sodium hyaluronate,the urinary frequency decreased significantly than before start of treatment (32.8 vs 18.5 times/24 h).The maximum urinary output increased significantly compared with the output before treatment (86.7 vs 151.9 ml).Pain was reduced significantly after treatment (8.7 vs 3.0).The O'Leary-Sant IC score and the QOL were significantly improved (30.0 vs 17.0,5.9 vs 2.4,respectively) (P =0.000).After the third treatment,the symptoms continued to improve.The treatment results were best in the fifth week at the time of the sixth injection of sodium hyaluronate.Symptoms rebounded at six months.However compared with that before treatment,the difference was still statistically significant ( P =0.000).ConclusionsBladder hydrodistention under anesthesia for severe intractable IC patients produces immediate effectiveness.Sodium hyaluronic infusion can alleviate urinary frequency and pain,and the effectiveness and duration of treatment are positively correlated.

BACKGROUND:There is no effective therapy for obliterative bronchiolitis after tracheal transplantation. A therapeutic strategy at microRNA (miRNA) molecular level plays a crucial role in the prevention and treatment of complications after organ transplantation.
OBJECTIVE:To analyze the miRNA differential expression profile in response to obliterative bronchiolitis after orthotopic tracheal transplantation in rats.
METHODS:The obliterative bronchiolitis model after lung transplantation was established through orthotopic tracheal transplantation in inbred strains of rats, and then was identified using histoIogical examination. Total miRNAs were detected by miRNA array and significantly differential expressed miRNAs were filtrated in the transplanted trachea tissues. The miRNA-146a, miRNA-155 and miRNA-451 with significantly differential expressions were used for relative quantitative study. Quantitative real-time reverse transcription-polymerase chain reaction was applied to verify the reliability of miRNA array results.
RESULTS AND CONCLUSION:The pathological examination showed that, obliterative bronchiolitis model in rats was successful y established at 4 weeks after orthotopic tracheal transplantation. A total of obliterative bronchiolitis-related 29 miRNAs were found in miRNA expression profiles, including 14 miRNAs with significantly down-regulated expression and 15 miRNAs with significantly up-regulated expression. Among them, the significantly up-regulated miRNAs (miRNA-146a and miRNA-155) and the significantly down-regulated miRNA-451 were involved in immuno-inflammatory reaction. The miRNAs play an important role in regulating pathophysiological changes of obliterative bronchiolitis after lung transplantation.