Objective To verify the targeting regulatory relationship between microRNA (miR)-21-5p and mitogen-activated protein kinase kinase 3 (MKK3).Methods miR-21-5p and MKK3 targeted relationship was analyzed by bioinformatics database predict.Constructing MKK3 3'UTR reporter vector (MKK3-3U) and mutation reporter vector (MKK3-3 U-M) was constructed by synthesizing miR-21-5p mimics (mimic),miR-2 1-5p inhibitor (inhibitor),nonsense miR (NC).NC (NC1 group),mimics (mimic1 group),inhibitor (inhibitor1 group) together with MKK3-3U cotransfected human embryonic kidney (HEK293) cells,respectively.Empty vector-pYr-MirTarget (NC2 group),MKK33U (Mt group),M KK3-3U-M (Mut group) together with mimic co-transfected HEK293,respectively.The fluorescence ratio was detected by dual luciferase assay.NC,mimics,inhibitor intervened human renal tubular epithelial (HK-2) cells,respectively.RT-PCR and Western blot were used to test cell MKK3 expression level in HK-2.Results 1.The fluorescence ratios of NC1 group,mimicsl group,and inhibitor1 group were 100.00％,67.99％,133.17％,and there was difference between mimic1 group and NC1 group(t =19.93,P ＜ 0.05).2.The fluorescence ratios of NC2 group,Mt group and Mut group were 100.00％,65.30％,98.48％,and there was difference between Mt group and NC2 group (t =14.39,P ＜ 0.05).There was no significant difference between NC2 and Mut group (t =0.63,P ＞ 0.05).3.MKK3 mRNA relative expression levels of the control group mimic2 group and inhibitor2 group were 1.36 ± 0.02,1.01 ± 0.04,1.43 ± 0.06 ; relative protein expression levels were 0.97 ± 0.05,0.62 ± 0.06,1.36 ± 0.32.MKK3 mRNA and protein levels of mimic2 group were significantly lower compared with the control group (F =85.98,118.26,all P ＜ 0.01).Conclusions MKK3 is the target gene of miR-21-5p.miR-21-5p can regulate the expression of MKK3 both in mRNA and protein levels.miR-21-5p may be an important regulatory molecule in p38 mitogen-activated protein kinase signaling pathway.

BACKGROUND: LIM Mineralization protein 1 (LMP-1), an intracellular non-secretory protein, plays roles in bone calcification. Presently, it is found that LMP-1 can promote an increase in bone morphogenetic protein 2 and transforming growth factor ? 1. This indicates that LMP-1 may recruit a mass of ossified factors to participate in the differentiation of osteoblasts. OBJECTIVE: To construct human LMP-1 gene adenovirus recombinant with AdEasy adenovirus vector system, and to detect LMP-1 expression in infected rabbit bone marrow mesenchymal stem cells (BMSCs). DESIGN, TIME AND SETTING: An opening experiment was performed at the Central Laboratory of South West Hospital, Third Military Medical University of Chinese PLA from March 2006 to February 2007. MATERIALS: Three New Zealand rabbits were used to isolate bone marrow mesenchymal stem cells. Plasmid pIRES2-EGFP-LMP-1 carrying human LMP-1 gene was kept in Department of Orthopedics, Second Hospital Affiliated to Chongqing Medical University. AdEasy was presented by Dr. Tong-Chuan He from USA. Human embryo kidney 293 cells were gifted by Wang from Department of Clinical Laboratory of Chongqing Medical University. METHODS: LMP-1 gene with a sequence encoding His-tag was amplified by using pIRES2-EGFP-LMP-1 plasmid as a template for polymerase chain reaction (PCR) with a specially designed downstream primer. The target gene was cloned to the pMD18-T vector for sequencing. Once verified, the gene was cut out by double endonucleases, connected to the shuttle vector pAdTrack-CMV. The newly constructed vector was linearized by PmeⅠ following efficient homologous recombination with the backbone vector pAdEasy-1 in BJ5183. The correct recombinant pAd-LMP-1 was linearized with Pac I and transfected to HEK293 cell by means of mediated Lipofectamine. The titer of virus was measured after amplification and purification. The mRNA and protein expression of LMP-1 was detected in BMSCs, which were infected with Ad-LMP-1 at the most appropriate MOI, were detected by reverse transcription (RT)-PCR and Western blot, respectively. MAIN OUTCOME MEASURES: Plasmid pAd-LMP-1 identification, its titre and efficiency of infection. mRNA and protein expression of LMP-1 were detected by RT-PCR and Western blot, respectively. RESULTS: The recombinant plasmid pMD18-T-LMP-1 carrying LMP-1 gene with His-tag was successfully constructed. After packaging and amplification of the recombinant adenovirus, the 3.5?109 efu/ml titer of Ad-LMP-1 was obtained by CsCl gradient purification. The optimal efficiency infection was 50%-70%, which was get after Ad-LMP-1 infected BMSCs for 3 days at the most appropriate MOI 150. The mRNA and protein expression of LMP-1 in infected BMSCs had been proved. CONCLUSION: The recombinant adenovirus containing human LMP-1 gene with His-tag is successfully constructed. The BMSCs infected with recombinant adenovirus Ad-LMP-1 can effectively express LMP-1.

It is reported that polyethylene glycol-lipid (PEG-lipid) derivatives increase liposomes stability, prolong the blood circulation of liposomes, enhance their tumor-targeting efficiency, and improve drug efficacy. Therefore, it is of great importance to investigate the influence of modified PEG-lipid derivatives on the physical, chemical, and biological characteristics of liposomes for the promotion of dealing with the existed problems, such as the accelerated blood clearance (ABC) phenomenon when repeated intravous injection at a certain time-interval, and developing novel targeted pharmaceutical preparations. In this review, the effects of modified PEG-lipid derivatives were summarized in many aspects. It indicats that the chemical bonds (amide, ether, ester, and disulfide) between PEG and lipid, as well as the species of lipids, such as the commonly used phosphatidylethanolamine, cholesterol, and diacylglycerol have substantial effects on the grafted liposomes stability in vitro and in vivo. Besides, the properties of lipids (the fatty acid chain length and saturation) and the groups (methoxy, carboxylic and amino) at the distal ends of the PEG chains were also considered to be important factors. In the end, the influence of the average molecular weight of PEG and the molar ratio of PEG-lipid derivatives in the total lipid were further focused.

Objective: To evaluate happiness and the mental health of elderly who participate morning exercise in Wuxi area. Methods: 161 elderly who participated morning exercise and 111 elderly from urban area of Wuxi were investigated with Memorial University of Newfoundland Scale of Happiness (MUNSH), Symptom Checklist 90(SCL-90), and general health status questionnaire. Optimal Scaling was used to study the relationship between Happiness and affecting factors. Results: The average level of happiness of the morning exercises groups was higher than that of control. Factors having influence on mental health included interpersonal relationship, children satisfaction rating, and attitude to retirement, house condition.Conclusions: Elderly participating morning exercise have better mental health status.

Objective To assess the left ventricular longitudinal shrinkage function in liver cirrhosis patients with two-dimensional speckle tracking imaging (2D-STI). Methods Echocardiography and Doppler echocardiography were performed in 34 patients with liver cirrhosis and 35 healthy subjects of corresponding ages. High frame rate two-dimensional images were recorded from apical long-axis view, four-chamber view and two-chamber view of left ventricle; then the left ventricular diameter, left atrium diameter, the peak filling velocity of E wave and A wave, E/A ratio, EF and FS were measured. The peak systolic strain of left ventricular segment was measured with two-dimensional strain software. Results Compared with healthy subjects, left ventricular diameter, left atrium diameter, EF and FS of liver cirrhosis patients were not statistically different (P>0.05), but the E/A ratio was lower (P<0.05). The peak systolic strain of most left ventricular segment in liver cirrhosis reduced significantly (P<0.05), except that of base segment of posterior wall, anterior wall, inferior wall, anterior and posterior interventricular septum, as well as middle segment of posterior interventricular septum. Conclusion The heart shape, systolic and diastole function of liver cirrhosis are abnormal. 2D-STI can early and accurately evaluate the systolic function of liver cirrhosis.

[Objective] The study was conducted to investigate the effect of micronized fenofibrate on acute insulin response in the subjects with impaired glucose metabolism and hypertriglyceridemia. [Methods] Fifty-three subjects were randomly (2:1 ratio) allocated to fenofibrate group (n=36, including IFG 3 cases, IGT 19 cases, IFG/IGT 6 cases, T2DM 8 cases) or control group (n = 17, including IFG 1 case, IGT 9 cases, IFG/IGT 4 cases, T2DM 3 cases) without any intervention for 3 months. Fasting blood samples were collected for measuring fasting plasma glucose (FPG), free fatty acids (FFA), and lipid profile. IVGTTs were carried out with measurement of plasma insulin before and after treatment. Acute insulin response (AIR), the maximum insulin concentrations (C_(INS,MAX)) to fasting insulin (FINS) ratio (C_(INS,MAX)/FINS) and values of the maximum insulin concentrations increment (△C_(INS)) during IVGTT were calculated as indexes of first-phase insulin secretion. HOMA insulin resistance index (HOMA IR) was used for assessing insulin resistance. [Results] After 3-month treatment, the lipid profile was evidently improved in fenofibrate group. Levels of trigiyceridemia (TG), low-density lipoprotein cholesterol and FFA were significantly reduced and high-density lipoprotein cholesterol increased significantly. Waist circumference was also significantly declined. No change of above indicators was found in control group. In fenofibrate group, C_(INS,MAX)/FINS and △C_(INS) were significantly increased (median 8.4 pmol/L vs. 5.3 pmol/L, 808±473 pmol/L vs. 660±472 pmol/L, both P<0.0001), along with great improvement of AIR (5 585±3 441 pmol·L~(-1)·min~(-1) vs. 4 444±3 642 pmol·L~(-1)·min~(-1), P<0.0001). The level of FINS and HOMA IR was also markedly reduced (108±65 pmol/L vs. 166±115 pmol/L, P = 0.002; 3.8±2.3 vs. 6.0±4.2, P = 0.001). In contrast, there were modest declining in acute insulin response (AIR: 4 313～1 943 pmol·L~(-1)·min~(-1) vs. 5 362±2 861 pmol·L~(-1).min~(-1); C_(INS,MAX)/FINS: median 4.6 vs. 7.0, P= 0.01; △C_(INS): 641±286 pmol/L, vs. 720±321 pmol/L, P= 0.003 9) and increasing HOMA IR (7.8±4.2 vs. 5.6±3.2, P<0.000 1) in control group after 3-month follow-up. The improvement of AIR was correlated with the decreasing of plasma FFA and TG (r=0.41, 0.36, P = 0.002, 0.014), but no correlation with the changing of FPG and HOMA IR. [Conclusions] These results indicated that sbort-term lipid-lowering treatment with fenofibrate evidently improved acute insulin response and alleviated insulin resistance in subjects with impaired glucose metabolism and hypertriglyceridemia. Moreover, the improvement of insulin secretion capacity may be mainly due to the relieving of iipotoxity resulting from finofibrate.

Thirty six patients with hypertriglyceridemia and impaired glucose regulation or newly diagnosed type 2 diabetes, whose fasting plasma glucose was ≤8.0 mmol/L, were treated by fenofibrate for 3 months. Lipid profile, insulin during intravenous glucose tolerance test and oral glucose tolerance test ( including glucose) were measured before and after treatment After treatment, lipid profile was significantly improved. Insulinogenic index (△I30/△G30) and acute insulin response were significantly increased (98. 9vs. 129. 2, 3558.9 vs. 4783. 3 pmol · L - 1 · min - 1, respectively, P ＜ 0. 05 ). Fasting insulin and insulin resistant index in homeostasis model assessment ( HOMA IR) decreased ( 128. 6 vs. 84. 8 pmol/L, 4. 8 vs.3.0, respectively, P ＜0. 05 ). The improvement of insulin secretory function was more significant in patients with higher triglyceride (TG ＞ 3. 3 mmol/L). These results indicate that short-term lipid-lowering treatment with fenofibrate can improve β-cell function and insulin resistance. Patients with higher triglyceride are likely to achieve more benefit from lipid-lowering treatment.

In recent years, the studies of the treatment of cerebral infarction have turned to the areas of neuroprotective agents. The promotion of angiogenesis after stroke will become one of the novel approaches after cerebral infarction. Studies have shown that the activation of the kallikrein-kinin system (KKS) after cerebral infarction may promote angiogenesis after cerebral infarction, improve the neurological deficits, and reduce the mortality and disability. This article reviews the regulatory mechanisms of KKS in the angiogenesis after cerebral infarction.

BACKGROUND:The anatomical strucure of acetabulum is smal and shal ow in adult acetabular dysplasia patients. The large amount of cal us and scar tissues in the acetabulum make it difficult to identify and instal the acetabular cup during arthroplasty. The comprehensive understanding of the acetabulum before arthroplasty is the premise for selecting the appropriate acetabular prosthesis and making the acetabular reconstruction program. OBJECTIVE:To evaluate the application value of three-dimensional reconstruction technique in choosing the size of acetabular cup before total hip arthroplasty for acetabular dysplasia. METHODS:Spiral CT was carried out in the 11 acetabular dysplasia patients who waiting for total hip arthroplasty. The acetabulum was multi-planar reconstructed, and the size of the acetabular cup was determined through digitized acetabular cup template implantation, and then the mathching degree assessment was performed to compare with the actual size. RESULTS AND CONCLUSION:Spiral CT could clearly show the acetabular morphology, and the 71.4%of the acetabular size chosen in the three-dimensional preoperative plan was the same as actual one, the intraclass correlation coefficient was 0.888. The agreement was much higher than that of two-dimensional preoperative plan based on X-ray plain film. For the patients with acetabular dysplasia, the acetabulum became saml er and shal ower, and there were various extents of bone defects in the superior-lateral acetabulum. Three-dimensional multi-planar reconstruction can effectively evaluate the acetabular morphology, and three-dimensional preoperative plan can provide useful information for the choice of implant.

Objective To establish and evaluate the animal model induced by inhalation injury of airborne fine particulate matter (PM2?5). Methods We manufactured equipment for rats aerosol inhalation with PM2?5. The effects of several facters such as concentrations(100 ± 10 μg/m3、150 ± 10 μg/m3、200 ± 10 μg/m3 )、time(1w、2w、4w、8w、12w)、method (non?exposed intratracheal instillation method and aerosol inhalation) and animals (Wistar rats, BN rats and guinea pigs) were investigated to establish the model. The respiratory rate, forced vital capacity ratio of forced expiratory volume ( FEV1/FVC) and arterial partial pressure of oxygen ( PO2 ) were measured, the pathological changes of bronchial and lung tissues under light microscope were observed. The success animal model was builded as the pneumonia was observed from the pathological changes of lung tissue. Results The Wistar rats exposed to PM2?5 aerosol inhalation for 8 weeks, we can see that the weight growth rate of rat decreased, WBC count and mononuclear cells count increased, the macrophages ratio decreased in BALF, the respiratory rate of lung increased while arterial PO2 and FEV1/FVC decreased, inflammation and pulmonary fibrosis changes were observed by bronch and pulmonary pathology, inflammatory changes with a dose?response relationship were observed. Exposed to PM2?5 aerosol inhalation for different time(1 w、2 w、4 w、8 w、12 w)with same dose, the score issue lesions of lung and bronchus in Wistar rats increased and the 8w group is obvious. The Wistar rats exposed to PM2?5 with different method ( aerosol inhalation and non?exposed intratracheal instillation method) for 8w, the aerosol inhalation worked as effectively as perfusion while mortality rate of aerosol inhalation is lower. Different animals ( Wistar rats, BN rats and guinea pigs) exposed to PM2?5 aerosol inhalation for 8w, the same results were observed with three method respectively while mortality rate of Wistar rats lower. Conclusions The optional conditions that the Wistar rats were continuously inhaled for 8w PM2?5 with a dose of 150 ± 10 μg/m3 were established. The animal model could be used on a national scale, especially in Fujian province. The results would be useful for the development of the research of the prevention and countermeasures of PM2?5 pollution.