AIM:To study the microRNA profiling in the serum of insulin-resistant mice and the mechanism of insulin resistance induced by related microRNAs.METHODS:A high-fat diet was used to induce insulin resistance model in KM mice.The microRNA profiling in serum of insulin-resistant and normal mice was analyzed by microarray chip and were validated by real-time PCR.miRanda data base was used to forecast target genes.miRBase was used to obtain the se-quences of related microRNAs, based on which protein interactions were predicted using the online analytical tool STRING. RESULTS:In serum of insulin-resistant mice, the expression of miR-125, miR-126, miR-143, miR-30a, miR-199a, miR-127, miR-184, miR-30e, miR-134, miR-195, miR-206, miR-429, miR-212, miR-362, miR-382, miR-154 and miR-466h was significantly up-regulated.miR-211, miR-504, miR-877 and miR-1930 were significantly down-regulated. miR-143 associated with insulin resistance was able to bind to 3'-UTR of fat mass and obesity-associated protein (FTO), and FTO was found to interact with Rpgrip1l, Tmem18, Mc4r, Npy, Hhex, Tcf712, Cdkal1, Slc30a8, Igf2bp2 and Tha-da.CONCLUSION:Twenty-one microRNAs in the serum of insulin-resistant mice induced by a high-fat diet are signifi-cantly different from those of normal mice, in which 17 kinds were significantly up-regulated.miR-143 closely related to in-sulin resistance is able to regulate FTO protein expression, which interacts with other 10 proteins associated with the occur-rence and development of diabetes.The results are also useful for further study of the molecular mechanisms in insulin re-sistance.

Objective:To screen the plasma microRNAs( miRNAs) of differential expression in a high fat diet-induced insulin resistance in mouse models;further investigations on the mice with insulin resistance treated by TLR4 inhibitors TAK-242,and to study the changes of plasma miRNAs expression profile and the relationship among TLR4, miRNAs and high fat diet-induced insulin resistance. Methods:The plasma samples were from 3 mouse groups of previous study,namely,the control group with general basic diet ( low fat diet,LFD) ,TLR4 inhibitors TAK-242 treatment group with a high fat diet ( HFD-T) and the high fat diet control group( HFD-C) . The differential expressed miRNAs was screened by expression profiling of plasma miRNAs, which was detected using mouse miRNA microarray. The quantitative Real-Time PCR ( qRT-PCR ) was used to verify the results of microarray. The target genes of differential expressed miRNAs were predicted in TLR4 signaling pathway using bioinformatics methods,and the GO and KEGG database molecular annotation system were used to investigate the main effects of the miRNAs targeted genes on the biological functions or signal pathway. Results:The screening results of miRNA microarray chip showed that,comparing miRNAs expression between HFD group and LFD control group,185 miRNAs were significant in the high fat diet group,including 6 up-regulated and 179 down-regulated miRANs. A significant difference of miRANs was also found between HFD-T group and LFD control group,the total number of differential expression miRNAs was 171,and all of them were down-regulated. Comparing miRNAs expression between HFD-C group and HFD-T group,13
miRNAs were significant in HFD-T group,all of them were down-regulated. Bioinformatics analysis results showed that a total of 10 in-teraction proteins with TLR4 were predicted;the difference of mmu-miR-3095-3p,mmu-miR-5113,mmu-miR-709 and mmu-miR-335-3p expression levels was more than 1 000 times between HFD-C group and HFD-T group,and their target genes can be found in TLR4-in-teraction protein or Toll like receptor signaling pathway;GO and KEGG analysis showed 74% of these target genes belonged to the biological processes genes, and the transcription factors accounted for 82%. The expression of mmu-miR-3095-3p, mmu-miR-5113, mmu-miR-709 and mmu-miR-335-3p detected by qRT-PCR exhibited the similar patterns of down regulation to those shown in microarray results. Conclusion:When insulin resistance occurs,there is a change in plasma miRNAs expression profile,this change is associated with TLR4 and its signaling pathways. The finding enrichs the possible mechanisms of insulin resistance and provides a basis for finding miRNAs diagnostic markers for early diagnosis of insulin resistance.

Performance evaluation system of 13 research institutions funded by the South Korean government was introduced in terms of policy and management.The situation of Korea Institute of Oriental Medicine performance evaluation in 2012 was presented.A reference has been provided for further improving science and technology evaluation system of China.

Objective To establish a high throughput screening assay for identifying human small molecular antagonists targeted IL-6R.Methods The full length gene of the human IL-6R extracellular region was amplified by PCR and cloned into a eukaryotic expression vector to construct recombination expression plasmid pABHis -IL6R that was then transfected transiently into HEK293T cells to prepare recombination protein IL-6R.Western blotting assay and receptor-ligand binding experiment were used to analyze the bioactivity of IL-6R.A new screening method based on ELISA was established using the function of IL-6R binding to its ligand and the characteristics of Fc fragment binding to IgG-HRP.Then Z′-factor was calculated and a known antagonist ab 47215 was used to assess the stability and reliability of the new assay .Results Recombination plasmid pABHis-IL6R was constructed and soluble IL-6R was prepared.IL-6R reported herein could be recognized by an anti-IL-6R antibody and specifically bind to its ligand in a dose response manner .A Z′-factor of 0.53 was obtained that could serve high throughput screening assay .Ab47215 , as a known specific antagonist , was able to block rhIL-6 from binding to the receptor in a dose-dependent manner in the new screening assay , the IC50 of which was (0.55 ± 0.11)μg/ml.Conclusion An innovative and easy screening assay for identifying human IL-6R antagonists is established , which might help discover potent and specific antagonists .

Objective To investigate cell-killing and bystander effects on gallbladder cancer (GBC-SD) cells transferred with double suicide genes (CD and HSV-tk). Methods CD and HSV-tk genes were transfected into PA317 cells using lipofectamine. GBC-SD cells were infected with viral supernatant. GBC-SD/CD+tk cells were treated with 5-FC and/or GCV. GBC-SD/CD+tk and GBC-SD were inoculated subcutaneously into the nude mice respectively and treated with GCV and 5-FC for 21 days. Results The killing effect of 5-FC and GCV in combination on GBC-SD/CD+tk cells is more effective than using 5-FC or GCV alone. The local bystander effect is significant while the distant bystander effect was not obvious. Conclusion The cell-killing efficacy of chemotherapy on GBC-SD transfected with double suicide gene was significantly increased, and the bystander effect was obvious.

ObjectiveTo investigate emergency diagnosis and treatment of pelvic fracture complicated with traumatic rupture of urethra and bladder,and to improve the success rate of treatment on pelvic fracture.MethodsClinical data of 52 cases of pelvic fracture complicated with traumatic rupture of urethra and bladder in department of emergency and urology from 2000 to 2010 was retrospectively analyzed.Results Among the 52 patients,there was 41 cases of pelvic fracture complicated with posterior urethral disruption,15 cases complicated with rupture of bladder and 4 cases complicated withtraumatic rupture of urethra and bladder at the same time.In 41 cases with posterior urethral rupture,6 individual's condition were relatively so severe that they onlyunderwent bladder puncture nephrostomy,and 29 cases underwent traction urethral realignment,the other 6 cases didn't undergo surgery; In 15 cases of patients with bladder rupture,2 patients were performed urethral realignment and bladder repair,11 patients underwent the bladder repair only and the other 2 patients were not performed surgery.There were 8 patients died and the mortality rate was 15.4％.Six died cases failed to conduct emergency surgery because of uncontrollable bleeding and another 2 cases died due to multiple organ failure.ConclusionPelvic fractures is a disease with more complications,it should be diagnosed as early as possible.Patients invalid for conventional anti-shock should be performed pelvic external fixation and emergency embolization to stop bleeding in the emergency department,and undergo associated processing after they are in stable condition.

Objective To study on the results of magnifying endoscopy in gastric atrophy, intestinal metaplasia (IM) and dysplasia, and evaluate their feasibility and accuracy for the diagnosis of these lesions. Methods One hundred patients were examined by magnifying endoscopy, Fujinon EG485 ZH modal, and stained with 0. 5% methylene blue. After defining magnifying endoscopic patterns of gastric pits as types A, B, C, D, and E, the diagnostic classification and endoscopic criteria were developed for the diagnosis of atrophy, IM and dysplasia. The results of 417 histopathological biopsy specimens taken from the corresponding areas of gastric mucosa under magnifying endoscopy were regarded as gold standard. Results Sparse and thick gastric pits mainly appeared in gastric atrophy, IM mainly appeared in gastric mucosa of type C, type D, and type E with positive stain, dysplasia appeared as depressed, slightly raised, or flat mucosa accompanied by loss of clear pattern, fine pits or coarse and irregular microstructure. The sensitivity and specificity of magnifying endoscopies in the diagnosis of atrophy, IM and dysphasia were 95. 85% , 95. 09% ; 88. 30% , 90.83% ; and 91.52% , 94. 41% respectively, all were higher than those of routine endoscopy. Conclusion The diagnostic accuracy significantly increased as depending upon the morphological features of gastric atrophy , IM, or dysplasia under magnifying endoscopy.

Objective To summarize the clinical experience and efficacy of surgical treatment for Stanford type A aortic dissection leading to acute lower limb ischemia.Methods From January 2014 to January 2018,12 patients with severe lower limb ischemia caused by acute type A aortic dissection were treated with Suns surgery.Among them,11 patients were treated with restoration of lower limb blood supply preferentially,including 10 cases of femoral artery bypass and 1 case of abdominal aorta-iliac artery stent graft implantation.Another case was treated with ascending aorta-femoral artery bypass after Sung surgery.Results 3 cases died of ischemia and necrosis of the lower extremities.Two of them died of multiple organ failure due to amputation and one died of low cardiac output due to refractory acidosis.Acute renal failure performed bedside CRRT in 5 patients and ECMO in 1 patient.The remaining 9 patients were discharged from the hospital and the symptoms of lower limb ischemia disappeared.After an average follow-up of 23 months,the re-examination of the aorta CTA showed that the bypass artery was unobstructed and the distal femoral artery was well developed.One patient infecting vascular prosthesis was cured by taking out the unit.Conclusion For acute lower limb ischemia caused by type A aortic dissection,blood flow of lower extremities should be restored as soon as possible to reduce mortality and complications.Femoral artery bypass and abdominal aorta-iliac arterial repair are simple and effective in reconstructing lower limb blood supply.

Objective To study whether portal vein thrombosis can be prevented by partial splenectomy combined with portoazygos devascularization,by preserving the integrity of the blood flow of the inferior splenic vessels.Methods 156 patients with portal hypertension and megalosplenia were divided into two groups.62 patients in group A who underwent partial splenectomy and 94 patients in group B who underwent total splenectomy.The postoperative complications,levels of WBC and PLT,volumes of residual spleen,hepatic vein pressure gradient (HVPG),and prevalence of portal vein thrombosis between the two groups were compared.Results The surgery time of group A was slightly higher than group B [(189.0± 38.5) vs.(128.0±36.3) min,P＜0.05].The blood loss of group A was slightly higher than that of group B [(328.0±68.9) vs.(294.0±49.1) ml,P＜0.05].The postoperative infection rate,change in HVPG and rehemorrhage rate between the two groups showed no significant differences (P＞0.05).The elevation in Leucocyte after operation of the two groups were the same,peaking on day 3.The platelet count of group A increased more gently than group B (2 weeks vs 1 week),and its peak was closer to the normal threshold (P＜0.05).The thrombosis rate of the portal vein system in group A was significantly lower than group B within 1 year (P＜0.01).The residual spleen did not enlarge after operation (P＞0.05).Conclusion Subtotal splenectomy combined with pericardial devascularization in patients with portal hypertension and megalosplenia preserved the blood flow of the subsplenic polar vessels and reduced the incidence of portal vein thrombosis.

Objective To study the impact of splenectomy and pericardial devascularization on the occurrence and development of portal vein thrombosis and the liver function in patients with cirrhosis complicated by portal hypertension.Methods 29 patients with cirrhosis and portal hypertension who underwent splenectomy and pericardial devascularization in the 302 Hospital of PLA from December 2012 to June 2013 were retrospectively studied.The incidences of PVT before and after operation were monitored.The liver function was assessed using the Child-Pugh classification.Results 29 patients with cirrhosis and portal hypertension underwent splenectomy and pericardial devascularization.The incidences of PVT in the preoperative period,12 days,3 months,6 months after operation were 10.3％,89.7％,51.7％,24.1％,respectively.The Child-Pugh scores in the preoperative period,12 days,3 months,6 months after operation were (5.2 ± 0.4),(5.6 ± 0.7),(5.2 ± 0.7),(5.3 ± 0.7),respectively.Conclusions The incidences of postoperative PVT increased after operation,but it decreased on long-term follow-up after operation.The liver function did not change.