Objective To evaluate the feasibility of transfection of Sonic hedgehog gene (SHH)into bone marrow mesenchymal stem cells(BMMSC).Methods After the SHH gene was transfected into BMMSC by electroporation apparatus,the transfection rate was evaluated by fluorescence inverted microscope.The growth curves of untransfected and transfected BMMSC were drawn,respectively,to observe the influence of transfection on cells.The expression of SHH gene in the BMMSC was detected by PCR,RT-PCR,Western-blot analyses.Results Through fluorescence inverted microscope,the observed transfection rate was appropriately 30％,PCR showed a obvious increase of SHH expression in transfected cells than that in untransfected cells,and it is quantified by qPCR for appropriately 7 times.Western-blot further demonstrated that the SHH protein expression in transfected cells had a distinct increase.However,it was observed that the exponential phase of BMMSCSHH growth curve delayed.The growth curves of both overlap 12 days after transfection.Conclusions This electroporation method can transfect exogenous SHH gene into BMMSC sufficiently with the effective protein expression in BMMSCSHH.It is the foundation of further research of genetic therapy for ischemic heart disease.

Objective To investigate the effect of Acanthopanax senticosus saponin (ASS)on the protein and mRNA expression of vascular endothelial growth factor (VEGF) in human HepG2. Methods HepG2 were incubated with ASS,the expression of VEGF was detected by ELISA assay,and the mRNA expression of VEGF was detected by RT-PCR assay. Results ASS (250,500 ?g/mL) decreased the expression of VEGF protein and VEGF mRNA (P

The therapeutic effects of intensive insulin therapy in treatment of traumatic shock combined with multiple organ dysfunction syndrome (MODS) were investigated. A total of 114 patients with traumatic shock combined with MODS were randomly divided into two groups: control group (n=56) treated with conventional therapy, and intensive insulin therapy group (n=58) treated with conventional therapy plus continuous insulin pumping to control the blood glucose level at range of 4.4-6.1 mmol/L. White blood cells (WBC) counts, prothrombin time (PT), serum creatinine (SCr), alanine aminotransferase (ALT), serum albumin and PaO(2) were measured before and at the day 1, 3, 5, 7 and 14 after treatment. The incidence of gastrointestinal dysfunction, the incidence of MODS, hospital stay and the mortality were also observed and compared. After intensive insulin therapy, the WBC counts, SCr, ALT and PT were significantly reduced (P<0.05), but the level of serum albumin was significantly increased (P<0.05) at the day 3, 5, 7 and 14. In the meantime, the PaO2 was significantly elevated at the day 3, 5 and 7 (P<0.01) after intensive insulin therapy. The incidence of gastrointestinal dysfunction, the incidence of MODS, the length of hospital stay and the mortality were markedly decreased (P<0.01). The results suggest early treatment with intensive insulin therapy is effective for traumatic shock combined with MODS and can decrease the length of hospital stay and the mortality.

Objective To explore the clinical value of high frequency ultrasound combined with serum tumor specific growth factor in diagnosis of thyroid cancer.Methods 155 patients with thyroid tumor received preoperative high frequency ultrasound and the detection of TSGF content,the histopathologic examination was given after operation.The clinical value of high frequency ultrasound and TSGF detection in diagnosis of thyroid cancer was analyzed.Results The sensitivity of high frequency ultrasound in diagnosis of thyroid carcinoma was 82.54％,the specificity was 81.52％.The sensitivity of TSGF in diagnosis of thyroid cancer was 79.37％,the specificity was 78.26％.The sensitivity of high frequency ultrasound combined with TSGF in diagnosis of thyroid cancer was 92.06％,the specificity was 93.48％.And the coincidence rate of combined detection was significantly higher than the two methods used alone,the difference was statistically significant (P ＜ 0.05).Conclusion High frequency ultrasound combined with TSGF detection can make up the shortage of the two methods,which has important value in improving the diagnosis of thyroid carcinoma.

Objective:To establish an HPLC method to determine the entrapment efficiency (EE) and drug loading (DL) of curcumin (CUR)and quercetin (QUE)loaded self-microemulsifying drug delivery system.Methods:A centrifugation method was used to isolate the free drug.The content of drug was determined by HPLC.The analytical column was a Purospher STAR LP C18 column (250 mm×4.6 mm,5 μm) and the column temperature was 30 ℃.The mobile phase was acetonitrile-4% acetic acid (50∶50) and the flow rate was 1.0 ml·min-1.The UV detection wavelength was set at 370 nm and the injection volume was 10 μl.Results:CUR and QUE were linear within the range of 10.728-96.552 μg·ml-1 (r=0.999 8) and 1.08-9.72 μg·ml-1 (r=0.999 9),respectively.The average recovery was 99.98%(RSD=1.46%,n=9) and 100.34%(RSD=1.06%,n=9),respectively.In CUR-QUE-SMEDDS,the EE of curcumin and quercetin was (95.97±0.50)% and (95.91±2.52)%,and the DL was (25.82±0.15) mg·g-1 and (1.80±0.05)mg·g-1,respectively.Conclusion:The method is accurate,rapid and simple,and suitable for the determination of DL and EE in CUR-QUE-SMEDDS.

The aim of the study was to prepare and evaluate the quality of negatively charged selfmicroemulsifying drug delivery system of curcumin (NC-CUR-SMEDDS).Based on the CUR-SMEDDS,the optimum amount of excipients was confirmed by the single-factor design experiment taking mean particle size,Zeta potential,drug entrapment efficiency and the drug loadings of curcumin as the evaluation indices for the preparation of NC-CUR-SMEDDS.The quality of NC-CUR-SMEDDS was evaluated by observing its appearance status,transmission electron microscope micrographs and determining particle diameter,Zeta electric potential,drug entrapment efficiency and drug loading.As a result,it was found that the Zeta potential reached -43.43 ± 0.29 mV when 4％ docusate sodium was added.The appearance of NC-CUR-SMEDDS remained clarified and transparent,and the microemulsion droplets appeared spherical without aggregation with uniform particle size distribution.The mean particle size was 14.08 ± 0.082 nm,the drug loading of curcumin was 26.48 mg·g-t and the drug entrapment efficiency was 94.12％.It was concluded that NC-CUR-SMEDDS with high entrapment efficiency and uniform particle size distribution met the requirement of inflammatory target binding in the colon.

This study was aimed to explore differential effects of various sections of the velvet antler on promoting cell proliferation in vitro. The NRK-49F cell line from rat kidney fibroblasts was used as the cell model. The cell proliferation rates of the water extracts from the upper, middle and lower section of fresh velvet antler were measured by the MTT method. BCA method was used in the detection of protein concentration. The SDS-PAGE method was used in the analysis of difference composition of the sample protein. The results showed that soluble protein content of the upper, middle and lower section were 17.89, 16.04 and 6.89 mg·mL-1, respectively. From the top to the base, the soluble protein content of velvet antler was decreased. After 24 h treatment, when the protein concentration of the upper and middle section samples of the velvet antler were 800 μg·mL-1 and 600 μg·mL-1, the cell proliferation promoting rates reached the maximum, which were 66.76% and 64.36%, respectively. And when the lower section sample of the velvet antler was 1 000 μg·mL-1, the cell proliferation promoting rates reached the maximum, which was 58.87%. After 48 h treatment, when the upper and middle section samples of the velvet antler were 800μg·mL-1, the cell proliferation promoting rates reached the maximum, which were 219.56% and 215.86%, respectively. And when the lower section sample of the velvet antler was 1 000 μg·mL-1, the cell proliferation promoting rates reached the maximum, which was 169.20%. The velvet antler on the proliferation of cells was much better than the 10% fetal bovine serum. The figure of SDS-PAGE showed the slight difference in the protein composition of three part of the velvet antler. It was concluded that all samples had promoting effects on cell proliferation with concentration-depen-dent, and the main protein in different part of the velvet antler had minor differences.

Objective To investigate the survival and differentiated situation of transplanted mesenchymal stem cells ( MSCs) transfected with Sonic hedgehog (SHH) gene after chronic myocardial infarction (MI).Methods MSCs were obtained from bone marrow of limb bones by density gradient centrifugation combined with attachment culture method .SHH gene was cloned and eukaryotic expression plasmid pcDNA 3.1-SHH was constructed .Then nucleofector TM was used to transfer SHH gene into MSCs and the expres-sions of mRNA and protein of SHH gene were detected in vitro .Myocardial infarction model was established by ligating the left anterior descending branch .A total of 150μl (3 ×106 ) of MSCs transferred with SHH gene was injected into ischemic area at the fourth week after MI model was set up .Brdu immunohistochemical staining and electron microscope examination were used to detect the survival and differentiation of MSCs with SHH in vivo at the 1st, 2nd, 4th, and 8th week after MSCs SHH transplantation.Results It was con-firmed that SHH gene clone and the recombinant eukaryotic expression vector pcDNA 3.1-SHH construction were successful by gene se-quence analysis and double digestions with EcoR I and Bamh I .Reverse transcriptase polymerase chain reaction ( RT-PCR) and West-ern-blot demonstrated that the expression of SHH mRNA was significantly increased in transfected MSCs compared with untransfected MSCs.Brdu immunohistochemical staining confirmed MSCs with SHH were survived from one to eight weeks in MI area after transplan -tation.Electron microscope examination showed that MSCs had the trend to be differentiated into myocardiac -like cells in MI area at the 4th and 8th week after transplantation .Conclusions The expression of SHH mRNA was significantly increased while MSCs were transfected with SHH gene , and MSCs SHH could survive and differentiate in good condition in the transplanted recipients .Those data provide the experimental basis for further transgenic cell therapy research of ischemic heart disease .

Objective To assess the series of special anti-fatigue foods including JuntiⅠ, JuntiⅡ, JuntiⅢ, and Jun-tiⅣon military physical performance capacity .Methods Fifty-four soldiers , selected from a border defense troop , were randomly designated to control group , trial group 1 and trial group 2.Subjects of two trial groups were supplied with No .1 nutritional package ( including JuntiⅠ, Ⅲand Ⅳ) and No.2 nutritional package ( including Junti Ⅱ, Ⅲand Ⅳ),re-spectively, while no additional nutritional supplements were added in control group .After 7 days’ supplementation, a hard military exercise was performed to induce fatigue and an increasing load test was used to assess physical activity .RPE scale, exhaustive time and time taken to reach the 75% maximal heart rate were recorded while serum markers , such as glucose, lactate, BUN, LDH,and CK, were detected after test .Moreover, serum lactate and fatigue recovery scale were determined on the evening of the same day and the next morning .Results Prolonged exhaustive time and time taken to reach the 75%maximal heart rate and elevated RPE scores at 6 min were detected in both two trial groups compared with the control group .Meanwhile , after the increasing load test , elevated glucose concentration and reduced lactate , BUN, LDH and CK were also observed in both trial groups .Moreover, serum lactate of both trial groups was quickly recovered on the evening of the same day compared with the control group , and the next morning , serum lactate was even much lower in trial groups than in control group .The fatigue recovery scores were higher in trial groups at both time points .Meanwhile, there was no difference of such indexes between the two trial groups .Conclusion Through the combination use , the series of special anti-fatigue foods, inclucling No.1 and No.2 nutritional packages , can significantly improve the soldiers′physi-cal performance capacity , delay the physical fatigue emergence , promote physical activity recovery and prevent military training injury.