Objective To study the effect of applying food composition table and foodstuff substitution method on nutritional status in patients with diabetic nephropathy and undergoing peritoneal dialysis.Methods Forty patients with diabetic nephropathy,treated with peritoneal dialysis for over three months was included in this study.Under the guidance of the food composition table and foodstuff substitution method for six months,nutritional status was compared before and after treatment. Results There was a significant difference in HGS,triceps skin foId thickness(TSF),mid-arm circumference(MAC),midarm muscle circumference(MAMC),Hb,albumin(Alb),serum crcatinine after 6 mortths of intervention compared with those before intervention. Conclusions Food composition table and foodstuff substitution method can obviously impwve the nutritional status for patients with diabetic nephwpathy undergoing peritoneal dialysis and increase the patients'diet compliance as well.

Objective:To explore the Wnt-β-catenin signal molecule expression level in acute kidney injury rats renal tissue. Methods:A total of 48 rats accorded to the random number table method were divided into normal control group ( Control group) ,sham operation group ( Sham group) and ischemia reperfusion group ( IRI group) ,each group with 16 rats,Control group was given normal fed,IRI group were established ischemia reperfusion injury rats model,Sham group opened dorsal skin and then wound was closed. 4 rats were sacrificed respectively at 1,3,5,7 d after surgery,the pathological changes of kidney tissue were observed in the three groups,and the serum creatinine,urea level was detected in the three groups. Wnt4,β-catenin,Lrp6 protein expression was detected in three groups of rats by Western blot,Wnt4,β-catenin,Lrp6 mRNA in renal tissues were detected by Real-time quantitative RT-PCR. Results:There was no significant difference in renal injury score and serum creatinine between Control group and Sham group (P>0. 05),IRI group kidney injury score and serum creatinine,urea first increased and then gradually decreased,which was the highest in 3 d after surgery,IRI group renal injury score and serum creatinine,urea were significantly higher than those in Control group and Sham group 1,3,5,7 d after surgery (P<0. 05). There was no significant difference in Wnt4,β-catenin,Lrp6 protein between Control group and Sham group (P>0. 05),IRI group Wnt4,β-catenin,Lrp6 protein gradually increased,which reached to peak at 5 d after surgery,and then gradually decreased,IRI group Wnt4,β-catenin,Lrp6 protein were significantly higher than those in Control group and Sham group 1,3,5,7 d after surgery(P<0. 05). There was no significant difference in Wnt4,β-catenin,Lrp6 mRNA between Control group and Sham group (P>0. 05),IRI group Wnt4,β-catenin,Lrp6 mRNA gradually increased,which reached to peak at 3 d after surgery,and then gradually decreased,IRI group Wnt4,β-catenin,Lrp6 mRNA were significantly higher than those in Control group and Sham group 1,3,5,7 d after surgery (P<0. 05). Conclusion: The expression of Wnt-β-catenin signal molecule are significantly increase in acute kidney injury,and the activation of Wnt-β-catenin signal pathway is involved in the repair process of renal tissue.

BACKGROUND:At present, the development of reprogramming technology provides a wide prospect for stem cellresearch. Through the ectopic co-expression of reprogramming factors, the somatic cells can be reprogrammed to a pluripotent state, termed as induced pluripotent stem cells, which can avoid the ethical controversy faced in the research and application of embryonic stem cells. Also, we can generate patient-specific and disease-specific induced pluripotent stem cells, which significantly decrease immuno-rejection. However, reprogramming technology faces some chal enges, such as low efficiency and safety.
OBJECTIVE:Based on the characteristics of induced pluripotent stem cells and the principles of reprogramming, to detail the progress in reprogramming technology from five aspects, including cellresources, carriers, transcription factors, microRNA and signal transduction pathway. 
METHODS:A computer-based online retrieval was performed to search papers published form January 1990 to April 2013 in VIP periodical ful-text database, Wanfang periodical ful-text database, CNKI periodical ful-text database, PubMed database and Springer database with key words of“reprogramming, induced pluripotent stem cell, signal transduction pathway, epigenetics, microRNA, transcription factor, vector, somatic cell, smal molecule compound, safety”both in Chinese and English. After excluding objective-independent papers, 67 papers were included for further analysis.
RESULTS AND CONCLUSION:By exploring different cellresources, different carriers, various combination of transcription factors, microRNAs or inhibition of the signal transduction pathways, the reprogramming efficiency and safety have been improved greatly. However, currently, induced pluripotent stem cells stil could not meet the requirement of clinical application. To achieve the clinical application of induced pluripotent stem cells, it is urgent to explore the mechanism of reprogramming, and to optimize the programming strategy.

Objective To investigate the effects of recombinant trichosanthin(rTCS) on methylation status and expression level of p27 gene in HeLa cells.Methods HeLa cells was treated by different concentration(20 ?g/mL,40 ?g/mL,and 80 ?g/mL) of rTCS for 48 h and then methylation-specific polymerase chain reaction(MSP) was used to detect the promoter methylation status of the p27 gene,real-time PCR was used to detect levels of p27 and DNMT1 mRNA,and Western blotting assay was used to detect expression level of p27 protein before and after treatment with rTCS.Results Low expression level and promoter methylation status of the p27 gene were detected in HeLa cells.Treatment with 40 ?g/mL rTCS totally demethylated p27 promoter.Treatment with 20 ?g/mL,40 ?g/mL or 80 ?g/mL rTCS resulted in a 2.22-,4.00-or 6.03-folds increase in p27 mRNA level,respectively,and also a great increase in p27 protein level.A high DNMT1 expression level was observed in HeLa cells and treatment with 40 ?g/mL rTCS resulted in a 78% decrease at the DNMT1 mRNA expression.Conclusion rTCS could reverse promoter hypermethylation and re-activate the expression of p27 gene by inhibiting DNMT1 expression in HeLa cells,which indicates its potential use in cancer therapy.

To investigate the correlation between p16 gene mutation and laryngeal cancer biological behavior as well as its prognosis in laryngeal cancer.Method:24 speciments of primary laryngeal cancer and 10 speciments with benign lesion in larynx were examined for mutations in exon2 of p16 by using PCR-SSCP silver stainning technique.Result:Mutations frequency of laryngeal cancer was62.5% (15/24).Nothing was found in 10 cases with laryngeal benign lesion.Conclusion:There is a strong correlation between p16 gene mutation and the biological behavior of chinese laryngeal cancer, such as histologic differentiation, invasion stage, and regional lymph nodes metastasis(P≤0.05).PCR-SSCP silver tainning technique is one of the most sensitive and simplest measure for detecting genetic mutation.It is worth using in clinical laboratory because of its readiness, repetition and lower cost.

OBJECTIVE To investigate the change of tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β),intercellular adhesion molecules(ICAM-1),matrix metalloproteinases 2 (MMP-2) and MMP-9 contents in cultured pulmonary microvascular endothelial cells (PMVECs) in rats after perfluoroisobutylene (PFIB) exposure. METHODS PMVECs were separated and purified using a modified method of implantation of pulmonary tissues. After identification,PMVECs were divided into the normal control group and the PFIB-exposed groups(n=3). The PFIB-exposed groups inhaled PFIB at the concentration of 200 mg · m-3 for 5 min in a flow-past header,while the normal control group were PMVECs in quiescent condition. The supernatants and lysates of PMVECs were harvested at 0.5,1,2,4 and 8 h,respec?tively, after execution. The contents of TNF-α,IL-1β,ICAM-1,MMP-2 and MMP-9 were measured by ELISA,and the activity of MMP-2 and MMP-9 was measured by gelatin zymography. RESULTS① According to the morphologic characteristics of cell growth and the expression of specificity antigens and the bind experiment of phytohemagglutinin,the cells separated and purified by modified method shared the characteristics of PMVECs.②TNF-αwas rapidly expressed by PMVECs at 0.5 h post PFIB stimulation and the maximum value was achieved at 2 h post PFIB stimulation(P<0.05). The newly synthesized TNF-α was slowly released out of the cells. The maximum TNF-α in the supernatant was achieved at 4 h post stimulation.③Within 2 h of stimulation,PMVECs synthesized a large amount of IL-1β and peaks at 2 h. However,IL-1βwas never released to the extracellular milieu.④The amount of ICAM-1 was rapidly synthesized by PMVECs after PFIB stimulation,but at a low level.⑤After stimulation with PFIB,MMP-2 in the supernatant of PMVECs culture was gradually increased,peaked at 2 h and then decreased subsequently. The biological activity of MMP-2 in the supernatant was also enhanced after PFIB stimulation. PFIB did not stimulate synthesis or secretion of MMP-9,indicating that PMVECs were not the main source of MMP-9 during PFIB inhalation-induced acute lung injury. CONCLUSION PFIB stimulates the surviving PMVECs to synthesize a large amount of TNF-α,IL-1β, MMP-2 and conjunctive ICAM-1.

BACKGROUND:Induced pluripotent stem cel technology have solved the contradiction between the ethics and immune rejection, and this high-efficient and safe technique is becoming the mainstream of today’s research. OBJECTIVE:To comprehensively review the safety and application of induced pluripotent stem cel s.METHODS:A computer-based online retrieval of PubMed and CNKI was performed to search relevant papers published from January 2006 to April 2016, with the key words of“induced pluripotent stem cel , reprogramming, clinical application, safety, transcription factor, disease mode”in English and Chinese, respectively. RESULTS AND CONCLUSION:In recent years, research on induced pluripotent stem cel s has attracted much attention from the scientific community and the medical community, and this technique has successful y gained induced pluripotent stem cel s and overcome the problems of immunity and ethics. However, it is limited to the theoretical and laboratory research due to the inability to solve the safety, efficiency and re-differentiation mechanism of induced pluripotent stem cel s. Therefore, we are faced with enormous difficulties and chal enges, which involve al aspects of basic research, including how to safely and effectively induce the differentiation of induced pluripotent stem cel s into the desired cel type and how to establish a suitable disease model as wel as a high-throughput drug screening platform.

Objective To explore the clinical effect of laparoscopic electrocoagulating hook'method in treatment of rectal cancer.Methods The retrospective cohort study was adopted.The clinical data of 237 patients with rectal cancer who were admitted to the Second People's Hospital of Guangxi Zhuang Autonomous Region from June 2004 to June 2009 were collected.Among 237 patients undergoing radical resection of rectal cancer,120 using laparoscopic electrocoagulating hook'method were allocated into the laparoscopic electrocoagulating hook'group and 117 using laparoscopic ultrasound knife's method were allocated into the laparoscopic ultrasound knife's group.Observation indicators included:(1)treatment situations:operation time,volume of intraoperative blood loss,number of lymph node dissected,time to anal exsufflation,time for out-of-bed activity,duration of hospital stay,treatment expenses and occurrence of complications(anastomotic leakage,intestinal obstruction,postoperative dysuresia and sexual dysfunction),(2)follow-up situations.Follow-up using outpatient examination and telephone interview was performed to detect tumor recurrence and metastasis up to December 18,2014.The colonoscopy,chest X-ray and abdominal computed tomography(CT)were applied to suspected patients with local or distal tumor recurrence.Measurement data with normal distribution were represented asx±s and comparison between groups was analyzed using t test.Count data were analyzed using chi-square test or Fisher exact probability.The survival curve was drawn by Kaplan-Meier method,and the survival rate was analyzed using the Log-rank test.Results(1)Treatment situations:all the patients underwent successful laparoscopic surgery,without conversion to open surgery.Operation time,number of lymph node dissected,time to anal exsufflation,time for out-of-bed activity and duration of hospital stay were respectively(156±33)minutes,18±2,(1.5±0.6)days,(1.5±0.5)days,(7.5±2.5)days in the laparoscopic electrocoagulating hook'group and(175±21)minutes,16±4,(1.6±0.5)days,(1.4±0.5)days,(8.0±1.8)days in the laparoscopic ultrasound knife's group,with no statistically significant difference in the above indexes between the 2 groups(t=25.324,20.117,21.477,1.617,1.770,P＞0.05).Volume of intraoperative blood loss and treatment expenses were respectively(116±12)mL,(2.4±0.3)×104 yuan in the laparoscopic electrocoagulating hook'group and(149±25)mL,(2.8±0.2)×104 yuan in the laparoscopic ultrasound knife's group,with statistically significant differences in the above indexes between the 2 groups(t=212.740,211.071,P＜0.05).Anastomotic leakage,intestinal obstruction and dysuresia or sexual dysfunction were respectively detected in 6,3,0 in the laparoscopic electrocoagulating hook'group and 7,3,5 in the laparoscopic ultrasound knife's group,with no statistically significant difference in the anastomotic leakage and intestinal obstruction between the 2 groups(x2=0.110,P＞0.05)and with a statistically significant difference in the dysuresia or sexual dysfunction between the 2 groups(P＜0.05).All the patients received postoperative chemotherapy of FOLFOX regimens for 6 cycles.(2)Followup situations:226 patients were followed up for a median time of 52 months(range,11-121 months),and 11 patients lost to follow-up were included in the study as censored data,including 5 in the laparoscopic electrocoagulating hook'group and 6 in the laparoscopic ultrasound knife's group.During the follow-up,67 patients were dead,including 58 died of tumor recurrence and metastasis,4 died of cerebral hemorrhage,3 died of myocardial infarction and 2 died of renal failure.The overall survival rate and overall survival time were 70.36％and 51 months.The overall local tumor recurrence rate,distal tumor recurrence rate and 5-year survival rate were 8.33％(10/120),25.83％(31/120),71.7％in the laparoscopic electrocoagulating hook'group and 7.69％(9/117),29.06％(34/117),72.4％in the laparoscopic ultrasound knife's group,with no statistically significant difference between the 2 groups(x2=0.033,0.310,0.002,P＞0.05).Conclusion The laparoscopic electrocoagulating hook'method in treatment of rectal cancer is safe and effective compared with traditional laparoscopic surgery,and it could also reduce the incidence of autonomic nerve-related complications and treatment expenses.

Objective:To observe the expression of IL-12 family subunit genes by real-time quantitative PCR in mice C6 glioma cells,construct the basis of the brain glioma research on IL-12 family in the future.Methods:Mice C6 glioma RNA was abstracted and reversed transcription cDNA.The mice C6 glioma cells mRNA expression influence of IL-12 family subunit genes was compared and analyzed by real-time quantitative PCR.Results: In mice C6 glioma cells, high expression abundances in IL-23a, IL-12a, midlde expression abundances in EBI3, IL-27, low expression abundance in IL-12b.Conclusion: IL-12 families are closely related to the occurrence and development of glioma,IL-12,IL-23 are regarded as the most potential anti-glioma cytokines among them,research de-velopments will bring a new way of brain glioma immune therapy.

BACKGROUND: Natural bilogical bone-derived materials processed with physical and chemical methods possess natural network pore system. They have good cellular compatibility, and help osteoblasts attach and grow on them, and can be used as the scaffolds of osteoblasts.OBJECTIVE: To observe the osteogenetic effect of partially deproteinised decalcified bone (PDDB) as scaffolds of osteoblasts in vivo.DESIGN: A randomized and controlled trial.SETTING: Central Laboratory of the First Hospital of Kunming Medical College.MATERIALS: Partially deproteinised decalcified bone; human embryonic periosteum-derived osteoblasts; twenty 4-week-old BALB/c nude mice,with the body mass of 25 to 28 g, of either gender.METHODS: This experiment was conducted at the Central Laboratory of the First Affiliated Hospital of Kuming Medical College from January to June 2003. PDDB composited with human embryonic periostea-induced osteoblasts were implanted into the nude mice after cultured for 1 week in vivo, 4 scaffolds in each nude mouse. Composite of scalfolds and cells implanted on the left side of the spine was set as experimental side and simple implanted material on the right side of the spine was set as blank control side. Then alkaline phosphatase activity and routine histological examination were performed 4 and 8 weeks after operation.MAIN OUTCOME MEASURES: General observation, alkaline phosphatase activity and routine histological examination were performed after the materials were taken out.RESULTS: Twenty nude mice entered the stage of result analysis. ① General observation of the implanted materials: No necrosis, ecpyesis, or fluidifying was found around the implanted materials, but ingrowth and enwrapption of soft tissues were found. ② Measurement of alkaline phosphatase activity: alkaline phosphatase activity after PDDB composited osteoblasts in vivo was stronger at week 8 than at week 4 [(22.854±6.018) nkat/g vs(11.286±4.268) nkat/g], and was much stronger than that of the simple implanted materials [(1.217±0.083) nkat/g vs (2.717±0.583) nkat/g]. ③ Results of routine histological examination: Cartilage formed at week 4 and part of cartilage formed new bone and marrow cavity at week 8 at the experimental side, cartilage and new bone formed much more as time went by, but there was no any cartilage or bone formation at the control side.CONCLUSION: Cartilage and bone form after PDDB composited with osteoblasts are implanted, and more cartilage and new bone form as time passes. PDDB can be used as the scaffolds of osteoblasts.