ObjectiveTo express and purify glutathione S-transferase(GST) and midkine(MK) fusion protein and prepare the ployclonsl antibody against MK.MethodsThe coding sequence of MK gene was amplified by RT-PCR from human gastric cancer tissue and inserted into pGEX-1λT vector.The recombinant vector was transformed into E.coli.BL21 ( DE3 ) to express the fusion protein GST-MK via IPTG induction.The expressed fusion protein was purified by GST affinity system and used to immunize rabbits for preparing the ployclonal antibody against GST-MK.ResultsThe expression vector pGEX-MK was constructed successfully.The expressed fusion protein was obtained by GST affinity system.The titer of the polyclonal rabbits antiserum was 1 ∶ 64 000 by indirect ELISA,Western blotting analysis showed that the antibody specifically bind to MK.ConclusionThe expressed protein and prepared polyclonal antibody provided useful reagents for MK further reaserch.

MicroRNA (miRNA) is a class of small ( about 22 nueleotides),single-stranded and noncoding RNA molecule.It is demonstrated that miRNAs participate in a wide array of physiologic processes,such as the development of early embryos and organ,cell proliferation,differentiation and apoptosis.MiRNAwhich has abnormal expression in hepatocellular carcinoma is closely related to the occurrence,developmentand metastasis of liver cancer.Liver cancer stem cells may be the root of cancer recurrence and metastasis.Researches have provided clear evidence that some kinds of miRNAs are important regulators in serf-renewal and differentiation of cancer stem cells.Therefore,regulating the expression of miRNAs in hepatoma cells and liver cancer stem cells may be a new approach to treat hepatocellular carcinoma or prevent the recurrence after treatment.

AIM　To build a model of two series of PPARγ agonists thiazolidinedione and aryketo-acid derivatives using 3D-QSAR method, and to reveal the structural features affecting the binding activity to PPARγ, which relates to antihyperglycemic and antihyperlipidemic activity and has a potential application to the treatment of type II diabetes. METHODS and RESULTS　48 agonists with selective activity for PPARγ were analyzed using CoMFA. Based upon the active conformation of rosiglitazone (BRL) extracted from its complex with PPARγ all agonists were aligned. The model from CoMFA showed a high ability to explain and predict the activity of PPARγ agonists with cross-validation correlation coefficient R2=0.656, that of non-cross-validataion R2=0.982, F10，37=201.1, and SE=0.115. CONCLUSION　The CoMFA contour map indicates that the steric fields mainly contribute to the binding effect, and especially a bulky group in the arylketo-acid series favors in the increase of affinity for PPARγ, as compared to the thiazolidinedione.

Aged ; Diagnosis, Differential ; Humans ; Intermittent Claudication ; diagnosis ; etiology ; Male ; Middle Aged ; Nervous System Diseases ; complications ; diagnosis ; Vascular Diseases ; complications ; diagnosis

ObjectiveTo observe the effect of auricular point sticking plus acupressure on blood pressure and lipid in hypertension patients with a dampness-phlegm constitution.MethodThe eligible subjects were randomized into antreatment group of 38 cases and a control group of 40 cases. The control group was intervened by conventional medication, while thetreatment group was additionally intervened by auricular point sticking plus acupressure. Before and after intervention, the blood pressure and lipid levels [total cholesterol (TC), triglyceride (TG), high density lipoprotein (HDL), low density lipoprotein (LDL)] were detected. ResultAfter intervention, the blood pressure, TC and TG levels were decreased in thetreatmentgroup (P<0.05,P<0.01), while the changes of HDL and LDL were statistically insignificant (P>0.05).ConclusionAuricular point sticking plus acupressure can somehow improve the blood pressure, TC and TG of hypertension patients with a dampness-phlegm constitution.

Objective To investigate the anti-infection and bone repair effects of composite scaffolds encapsulated with vancomycin and bone morphogenetic protein-2 (BMP-2).Methods Chitosan (CS) solution and nano-hydroxyapatite (n-HAP) mixed in a ratio of 1∶1 were made into solid materials by freeze drrying technology,and immerged into BMP-2/polylactic acid (PLA) and vancomycin/PLA microsphere solution to fabricate drug-loaded scaffolds.Efficiency of drug release was measured.After the induction of chronic infectious tibial bone defect,60 New Zealand white rabbits were assigned to isolate debridement group (Group Ⅰ),n-HAP/CS scaffold group (Group Ⅱ),n-HAP/CS + vancomycin/PLA group (Group Ⅲ) and n-HAP/CS + vancomycin/PLA + BMP-2/PLA group (Group Ⅳ) according to the random number table,with 15 rabbits per group.After 8 weeks of treatment,X-ray filming,Norden score,HE staining and narrow culture were performed to evaluate the anti-infection ability and bone repair ability in each group.Results Pore size of the n-HAP/CS scaffold constructed in this study ranged from 80 to 270 μm (mean,146.53 μm).Biomechanical test showed the elastic modulus of the scaffold material was (27.50 ± 1.58)MPa and the compressive strength was (0.75 ± 0.04)MPa.BMP-2/PLA and vancomycin/PLA microspheres into ball shape uniformly distributed on the scaffold pores.Drug-loaded scaffold showed relatively stable release of vancomycin and BMP-2 in a period of 18 days.With the extended time amount of the drug release gradually reduced.On the X-ray films 8 weeks after the treatment,sinus formation was visible in groups Ⅲ and Ⅳ,and emergence of periosteal reaction with focal low density shadow was demonstrated in groups Ⅰ and Ⅱ.Norden scores of groups Ⅰ,Ⅱ,Ⅲ and Ⅳ were (2.64 ±0.33) points,(2.63 ±0.36) points,(1.14 ±0.29) points and (0.89 ±0.42) points respectively,indicating group Ⅳ ranked the lowest compared with other three groups (P＜0.05) and group Ⅲ showed significant difference in comparison with groups Ⅰ and Ⅱ (P ＜0.05).HE staining showed massive infiltration of inflammatory cells and destroyed bone trabeculas in groups Ⅰ and Ⅱ,trabecular bone arranged neatly in groups Ⅲ and Ⅳ,and thicker trabecular bone in group Ⅳ than in group Ⅲ.Marrow culture showed obvious pale yellow colony formation in groups Ⅰ and Ⅱ,but no colony formation in groups Ⅲ and Ⅳ.Conclusions Vancomycin/BMP-2-loaded tissue engineering scaffolds are constructed successfully.Drug-loaded n-HAP/CS composite scaffold is identified as an ideal implant material for treatment of chronic osteomyelitis in rabbits

ObjectiveThe purpose of this study was to determine whether fQRS is associated with increased ventricular arrhythmic event in patients with Acute ST- elevation Myocardial Infarction.Methods 12-lead ECG, 24h-Holter monitoring and biochemical markers detection were performed in sixty cases with acute ST-elevation myocardial infarction, which were divided into two groups[fQRS group and nonfQRS group]according to Fragmented QRS (fQRS) on 12-lead ECG.Then ventricular arrhythmic events were analyzed.ResultsVentricular premature beats (24 h, 3140 +555 vs 1226 + 142, P ＜0.01) and the incidence of complex ventricular arrhythmic events(34.4％ vs 10.7％, P ＜0.05) in fQRS group were significantly higher than that in non- fQRS group.ConclusionsFragmented QRS may be used to predict ventricular arrhythmic events in patients with acute ST- elevation myocardial infarction.

BACKGROUND:Fracture of the distal humerus is a common type of fracture. Conservative treatment or metal implant fixation can be used during the clinical treatment of elderly patients, but the choice of specific treatment options stil lacks of clear conclusion. OBJECTIVE:To explore the clinical repair effect and biocompatibility of titanium aloy implant internal fixation for aged patients with distal humeral fractures. METHODS: A total of 41 elderly patients with humeral fractures, who were hospitalized in Danzhou Municipal First People’s Hospital from July 2011 to July 2014, were enroled in this study. In accordance with the wishes of patients, repair program was selected. 22 patients in the control group received conservative treatment. 19 patients in the observation group received titanium aloy implant fixation. After 12 months of folow-up, elbow joint activity, elbow joint function recovery and adverse reactions were observed and compared in the two groups. RESULTS AND CONCLUSION: After conservative treatment and internal fixation, each index score and total score of range of motion of shoulder joint were significantly higher in the two groups as compared with pre-treatment (P < 0.05). Each index score and total score were significantly higher in the observation group than in the control group (P < 0.05). Curative effect evaluation results showed four excelent patients and three good patients in the control group, eight excelent patients and four good patients in the observation group. The excelent and good rate was significantly lower in the control group than in the observation group (P < 0.05). Two patients suffered from mild inflammatory hyperplasia during taking out the fixator in the observation group, and the remaining patients did not affect inflammatory hyperplasia or capsular tissue. In the observation group, after surgery, one case experienced numbness of the ulnar nerve, which disappeared after treatment with trophic nerve. One case suffered from wound infection, which was controled by antibiotic treatment, and did not induce a serious outcome. After folow-up, none of them had nonunion or internal fixation loosening. The results show that titanium aloy implant fixation for the treatment of distal humerus fractures can get better repair effect compared with conservative treatment, can effectively improve the elbow joint function, and titanium aloy material has good biocompatibility.

Humans ; Hyperesthesia ; diagnosis ; etiology ; Pharynx ; physiopathology ; Syndrome

Alternative splicing of pre-messenger RNA (pre-mRNA) is a crucial mechanism for the diversity of the human transcriptome and proteome. Alternative splicing is a complex gene regulation process. Whole-transcriptome analysis shows that 95% of human exonic genes are alternatively spliced, involving various cis-acting elements and trans-acting factors. Any changes in any component or step may cause erroneous splicing events and lead to the occurrence of various related diseases. In addition to gene replacement therapy that directly changes the splicing results, RNA splicing modification is expected to become a new therapeutic strategy to alleviate or treat diseases by targeting and correcting abnormal pre-mRNA splicing. Splicing modification tools currently developed including RNA trans-splicing, antisense oligonucleotides, small interfering RNA, and small molecule drugs can correct abnormal splicing through different ways. This article reviews the resent progress of epigenetic regulation of pre-mRNA alternative splicing in recent years, and discusses the occurrence and regulation of alternative splicing, the types of diseases caused by related splicing defects, and the current-used tools for targeting and altering splicing. The importance of splicing modification strategies in the future treatment of human diseases is envisioned.