ObjectiveTo investigate the effect of mild hypothermia on neuron specific enolase (NSE) and superoxide dismutase (SOD) in serum of patients with large-area cerebral infarction.Methods160 cases with large-area cerebral infarction were divided into the treatment group and control group with 80 cases in each group. Patients of the control group were treated with routine therapy. Those of the treatment group were added with mild hypothermia therapy (MHT). The scores of NIHSS were assessed and NSE and SOD in serum were examined before treatment and 7, 14 and 30 days after treatment in two groups.ResultsThe NIHSS score of the treatment group was significantly lower than that of the control group ( P<0.05), and NSE level decreased, SOD vitality increased in the treatment group. Other indexes such as respiratory, pulse, serum kalium and etc of two groups were not different ( P>0.05).ConclusionMHT can improve he nerve function of patients with large-area cerebral infarction recovering and improve prognosis.

Objective To explore the effects of miRNA-218 on renal cell carcinoma of nude mice in vivo.Methods The pcDNA3.1-miR-218 and its control negative plasmids were stably transfected into renal cell carcinoma cell line A498 and 769-P.These cells were inoculated into nude mice in different groups to observe the changes of body and tumor and to detect the expression of miR-218 in the tissues of nude mice.Results In the A498 cells + pcDNA3.1-miR-218 transfected group,the weight loss of tumor bearing nude mice after 25 days was lower than that in the control group,and the tumor volume was smaller than that in the control group after 10 days (P ＜ 0.05).In the 769-P cells + pcDNA3.1-miR-218 transfected group,the weight loss of tumor bearing nude mice was lower than that in the control group after 19 days,and the tumor volume was smaller than that in control group after 10 days (P ＜ 0.05).The expression of miRNA-218 in bearing nude mice with A498 cells or 769-P cells transfected by miRNA-218 was significantly higher than that in the control group,and the difference was statistically significant (P ＜ 0.05).Conclusion Up-regulation of miRNA-218 expression can inhibit the growth of renal cell carcinoma of nude mice in vivo.

Objective To construct a stable expression plasmids miR-218,to lay the experimental foundation for the expression and mechanism of miR-218 in human renal cell carcinoma.Methods The primers were designed by Has-miR-218 and the miR-218 fragment were amplified by PCR,and then which was connected with the carrier pcDNA3.1 (+) to build a stable expression plasmids.The recombinant plasmids were trasfected into renal cell carcinoma cell line 769-P,and the expression of miR-218 in these cells were detected.Results The plasmids were constructat successfully,which was determind by enzyme digestion and sequencing results.The constructed expression plasmids pcDNA3.1-miR-218 transfection of human renal cell carcinoma cell line 769-P,miR-218 cxpression level of cells was significantly increased after trasfected by recombinant plasmids carrying miR-218 gene (relative expression quantity was 1.64).Conclusion The miR-218 expression plasmids pcDNA3.1-miR-218 were successfully constructed,the plasmids can be applied to the study of miR-218 function and mechanism in renal cell carcinoma.

The amount of the research-oriented graduates is increasing in affiliated hospitals of medical universities.To create an effective training model can not only help students to succeed but also contribute great to the research level of hospitals.In this study,we summarized our experience,such as individualized teaching,the unity of thinking and performing,words and deeds as well as unity of clinic and research on the formation of research ideas,attitudes and styles.Based on our experiences we hope to provide meaningful methods for the training of medical graduates.

BACKGROUND: Anti-tuberculous chemotherapy is the main method for treating bone and joint tuberculosis. However, systemic administration hardly maintains the effective drug concentration in the focus area, and the therapeutic efficacy is unsatisfactory. OBJECTIVE: To prepare a chitosan-gelatin/poly(lactic-co-glycolic acid) combined with drug-loaded hydrogel, which can release anti-tuberculosis drugs in situ for a long time and promote osteogenesis. METHODS: Isoniazid, a hydrophilic anti-tuberculosis drug, and a hydrophobic stromal cell derived factor-1 were loaded into poly(lactic-co-glycolic acid) by double emulsion method to prepare drug-loaded poly(lactic acid co-glycolic acid) microspheres, which were then mixed into chitosan gelatin/poly(lactic acid co-glycolic acid) combined with drug-loaded hydrogel. The ability of drug delivery and anti-tuberculosis of poly(lactic acid co-glycolic acid) microspheres and chitosan gelatin/poly(lactic acid co-glycolic acid) combined with drug-loaded hydrogels in vitro were tested. MC3T3-E1 cells were inoculated on the surface of microspheres and hydrogel respectively. The biocompatibility was detected by cell counting kit-8 assay. The osteogenetic activity was detected by alkaline phosphatase activity. RESULTS AND CONCLUSION: (1) The burst release of isoniazid in the microspheres was about 23.3% in 1 hour, 42.6% in 2 days, and then it entered the sustained-release stage in the later 25 days. The burst release of stromal cell derived factor was about 19.8% in 1 hour, 44.7% in 2 days, and then it entered the sustained-release stage in the next 25 days. The release of isoniazid and stromal cell-derived factor in the combined drug-loaded hydrogel was 8.3% and 8.5% in the first hour, respectively. The cumulative release rates on the second day were 15.2% and 17.6%, respectively, which were much lower than that of poly(lactic acid co-glycolic acid) microspheres. (2) After 4 weeks in vitro, the antibacterial diameter of the combined drug-loaded hydrogel was much larger than that of the drug-loaded microspheres, and the antibacterial rate was higher than that of the drug-loaded microspheres (P < 0.05). (3) The combined drug-loaded hydrogel and the drug-loaded microspheres had good cytocompatibility and cell viability was about 100%. (4) After 5 and 10 days of culture, there was no significant difference in the activity of alkaline phosphatase on the surface of drug-loaded hydrogel and drug-loaded microspheres. (5) These results show that the in situ chitosan-gelatin/poly(lactic acid co-glycolic acid) combined with drug-loaded hydrogel can be used for treating tuberculosis and other bone and joint infections.

Objective To observe the changes of intracerebral amino acid transmitters in the periventricular leukomalacia (PVL) of newborn rats with microdialysis and so as to explore the role of excitotoxicity in PVL.Methods Replicated the model for PVL at the age of postnatal day 5 (P5) by intracerebral injection of 3-nitropropionic acid (3-NPA).Before injection of 3-NPA,and 15 min,30 min,45 min,60 min,75 min,90 min after injection of 3-NPA,collected the sample of extracellular fluid (ECF) at the corpus callosum above the left ventricle through microdialysis,respectively.After microdialysis,the experimental rats were allowed to survive to P6-P14,and then they were killed and the brains were prepared for HE stain.The amino accids of dialysate were quantified through high performance liquid chromatography(HPLC),and then the excitotoxic index (EI) was calculated.Results Fifteen min to 45 min after injection of 3-NPA,the concentrations of glumate (Glu) and aspartic acid (Asp) of ECF elevated significantly,and then returned to the normal levels.Fifteen min to 75 min and 15 min to 30 min after injection of 3-NPA,the concentrations of glycine (Gly) and GABA significantly elevated,respectively,and returned to normal levels at 90 min and 45 min after injection of 3-NPA,respectively.But the EI,which indicated the balance of excitatory amino acids(EAAs) and inhibitory amno aciols(IAAs),significantly elevated 15 min to 75 min after injection of 3-NPA,then retured to normal level after 90 min.Sub-cortical and periventricular white matter rarefaction and significant lateral ventricle enlargement were observed in HE staining.Conclusions Changes of intracerebral amino acid transmitters in the PVL of newborn rats show regularity:EAAs,IAAs of ECF and EI elavate in the early stage,and then return to the normal level quickly.It indicates that excitotoxicity play a great role in PVL,especially at the early stage.Therefore,the preventions of PVL must be executed at the early stage.

Objective To evaluate the effect of hydrogen-rich saline on the expression of phosphor-p38 mitogen-activated protein kinase (p-p38MAPK) during cerebral ischemia-reperfusion (I/R) in rats.Methods Seventy-two adult male Sprague-Dawley rats,weighing 220-250 g,were randomly divided into 3 groups (n =20 each) using a random number table:sham operation group (group S),I/R group and hydrogen-rich saline group (group I/RH).Cerebral ischemia was induced in chloral hydrate-anesthetized rats by 2 h middle cerebral artery occlusion in I/R and I/RH groups.The artery was only exposed but not occluded in group S.At 3 days before operation and immediately after onset of reperfusion,hydrogen-rich saline (0.6 mmol/L) 10 ml/kg was intraperitoneally injected in group I/RH,while the equal volume of normal saline was given in S and I/R groups.Neurological deficits were blindly assessed and scored at the end of 24 h reperfusion.The animals were then sacrificed,and brains were removed for microscopic examination and for determination of the cerebral infarct size (by TTC),brain water content,cell apoptosis (by TUNEL),and expression of p38MAPk and phosphor-p38MAPK (p-p38MAPK) (by immunohistochemistry and Western blot).Apoptosis index was calculated.Results Compared with group S,neurological deficit score,apoptosis index,brain water content and cerebral infarct size were significantly increased,and the expression of p38MAPK and p-p38MAPK was up-regulated in I/R and I/RH groups.Compared with group I/R,neurological deficit score,apoptosis index,brain water content and cerebral infarct size were significantly decreased,and the expression of p38MAPK and p-p38MAPK was down-regulated in group I/RH.The pathological changes of cerebral tissues were significantly attenuated in group I/RH as compared with group I/R.Conclusion Hydrogen-rich saline can reduce cell apoptosis through inhibiting p-p38MAPK expression,thus attenuating cerebral I/R injury in rats.

Animals ; Antineoplastic Agents ; therapeutic use ; Apoptosis ; Apoptosis Regulatory Proteins ; metabolism ; Autophagy ; physiology ; Beclin-1 ; Humans ; Membrane Proteins ; metabolism ; Neoplasms ; drug therapy ; metabolism ; pathology ; Signal Transduction ; TOR Serine-Threonine Kinases ; metabolism ; Tumor Suppressor Protein p53 ; metabolism

OBJECTIVE:To establish the method for the detection of the known glucagon-like peptide 1 receptor (GLP1R) gene mutation site rs3765467(NT_007592.16,position:39065819),and to evaluate its accuracy and practicability. METHODS:Peripheral venous blood samples of 72 healthy subjects were collected in medical examination center of our hospital during Oct. 2015-Feb. 2016. The whole blood DNA was extracted by column extraction method. After amplified by touch down PCR,high reso-lution melting(HRM)method was adopted to analyze amplified product. Sequencing verification by double stranded chain termina-tion method(Sanger sequencing method)was performed for 38 test samples. The results of 2 methods were compared. RESULTS:The results of mutation scanning showed that there were 39065817 and 39065819 polymorphism sites in amplified segments. Four types of mutations were detected by HRM method [GCG/GCG,GCA/GCG or ACG/GCG,GCA/GCA or ACG/ACG,A(G) CA(G)],but 6 types of mutations was detected by Sanger sequencing method [GCG/GCG,ACG/GCG,ACG/ACG,A(G)CA (G),GCA/GCG,GCA/GCA]. CONCLUSIONS:HRM method can identify GCG/GCG and A(G)CA(G)genotype,but can not identify GCA/GCG and ACG/GCG heterozygous mutation,GCA/GCA and ACG/ACG homozygous mutation. The method is not suitable for the detection of single nucleotide polymorphism for multiple neighboring sites. In the detection of single nucleotide mu-tation,economical and simple method should be selected after comprehensive analysis of sequence.

Objective To evaluate the effect of hydrogen on the postoperative cognitive function in aged mice.Methods Sixty aged male Kunming mice,aged 12-14 months,weighing 35-45 g,were assigned into 3 groups (n=20 each) using a random number table:sham operation group (group Sham),partial hepatectomy group (group PH) and partial hepatectomy plus hydrogen-enriched saline group (group PH-H).On preoperative day 3,during operation and on postoperative day 3,hydrogen-enriched saline 10 ml/kg was intraperitoneally injected once a day in group PH-H,and the equal volume of normal saline was intraperitoneally injected once a day in S and PH groups.The spatial probe test was performed on postoperative day 7 to evaluate the cognitive function.Ten mice were selected on postoperative days 3 and 7 (after the end of the spatial probe test),and blood samples were collected for determination of serum tumor necrosis factor-alpha (TNF-α),interleukin-lbeta (IL-1β) and high-mobility group box 1 protein (HMGB1) concentrations by enzyme-linked immunosorbent assay.The animals were sacrificed after blood sampling,brains were removed and hippocampi were isolated for detection of the expression of TNF-α,IL-1β and HMGB1 by Western blot.Results Compared with group S,the percentage of time of staying at the target quadrant and the number of crossing the platform within 1 min were significantly decreased,the serum concentrations of TNF-α,IL-1β and HMGB1 were increased,and the expression of hippocampal TNF-α,IL-1β and HMGB1 was up-regulated in group PH (P＜0.05),and no significant change was found in the variables mentioned above in group PH-H (P＞0.05).Compared with group PH,the percentage of time of staying at the target quadrant and the number of crossing the platform within 1 min were significantly increased,the serum concentrations of TNF-α,IL-1β and HMGB1 were decreased,and the expression of hippocampal TNF-α,IL-1β and HMGB1 was down-regulated in group PH-H (P＜0.05).Conclusion Hydrogen can improve the postoperative cognitive function in aged inice.