Objective To systematically compare the operative and non-operative treatments of acute injury to the lateral ankle ligaments in terms of efficacy and complications.Methods The databases of MEDLINE (From January 1966 to October 2011 ),EMBASE (From January 1988 to October 2011 ),CBMdisc (From January 1978 to October 2011) and CNKI (From January 1980 to October 2011 ) were researched for eligible randomized-controlled trials (RCTs),controlled clinical trials,quasi randomized controlled trials and cohort study which compared operative and non-operative treatments for acute injury to the lateral ankle ligaments.The methodological quality of the eligible studies collected was evaluated.The data of the studies included were extracted for a Meta analysis to compare the motional recovery,functional stability,re-injury,residual pain and post-treatment complications of the ankle between operative and non-operative treatments.RevMan 5.0 software was used for statistical analysis.Results In all the eligible 13 studies included,713 patients were treated operatively and 817 patients non-operatively.The functional stability of the ankle was significantly better in the operative treatment group than in the non-operative treatment group [ OR =0.72,95％ CI,(0.52,0.99),P ＜ 0.05].The incidence of ankle arthrocleisis in the operative treatment group was significantly higher than in the non-operative treatment group [ OR =3.41,95％ CI (1.56,7.44),P =0.002].There were no statistical differences in the motional recovery [ OR =1.14,95％ CI (0.58,2.21),P＞ 0.05],incidence ofre-injury [OR=0.68,95％ CI (0.35,1.31),P＞0.05],residual pain [ OR =0.81,95％ CI(0.56,1.16),P＞ 0.05],or ankle dyskinesia [ OR =2.38,95％ CI (0.91,6.25),P＞ 0.05]between the 2 groups.The incidences,scar tenderness [01R=7.46,95％CI(1.32,42.08),P ＜0.05]and sensory nerve loss [OR=12.16,95％ CI(2.24,66.02),P ＜0.05]were significantly higher in the operative treatment group than in the non-operative treatment group.The total rate of complications was significantly higher in the operative treatment group than in the non-operative treatment group [0R=6.20,95％ CI (2.67,14.41),P ＜0.05].Conclusions Compared with non-operative treatments,operative treatments for acute injury to the lateral ankle ligaments can significantly improve the functional stability of the ankle,but make no significant differences in the motional restoration,re-injury or the residual pain of the ankle.In addition,operative treatments may increase the risk of complications.

Objective To evaluate the effects of dexmedetomidine used to supplement analgesia with sufentanil on the stress response and inflammatory response after cardiac valve replacement with cardiopulmonary bypass (CPB).Methods Forty-four patients of both sexes,aged 18-64 yr,with body mass index of 18-30 kg/m2,of American Society of Anesthesiologists physical status Ⅱ or Ⅲ (New York Heart Association Ⅱ or Ⅲ),scheduled for elective cardiac valve replacement with CPB,were randomly divided into sufentanil analgesia group (group S,n =20) and dexmedetomidine supplementing sufentanil analgesia group (group DS,n=24) using a random number table.After induction of general anesthesia,the patients were endotracheally intubated and mechanically ventilated.Combined intravenous-inhalational anesthesia was used to maintain anesthesia.Dexmedetomidine was infused at 0.3 μg · kg-1 · h-1 from closure of the chest to the end of surgery in group DS,while the equal volume of normal saline was given instead in group S.The patient-controlled intravenous analgesia (PCIA) was used for postoperative analgesia.PCIA solution contained sufentanil 2 μg/kg in 100 ml of normal saline in group S,or sufentanil 2 μg/kg plus dexmedetomidine 3 μg/kg in 100 ml of normal saline in group DS.The PCIA pump was set up with a 0.5 ml bolus dose,a 5 min lockout interval and background infusion at a rate of 1.5 ml/h.Visual analogue scale score was maintained ≤ 3.When visual analogue scale score ≥ 4,morphine 0.1 mg/kg was injected intravenously.The occurrence of analgesics-related respiratory depression,bradycardia and hypotension was recorded.At the end of surgery (T0),and 12,24,48 and 72 h after surgery (T1-4),blood samples were collected from the central vein for determination of serum cortisol,β-endorphin,C-reactive protein,interleukin-2 (IL-2),and IL-6 concentrations by enzyme-linked immunosorbent assay.Results No analgesics-related adverse events were detected in the two groups.Compared with group S,the requirement for morphine was significantly decreased,the serum cortisol concentration was decreased at T2-4 and the serum β-endorphin,C-reactive protein concentrations were decreased at T1-4,the serum IL-2 concentrations were increased at T1-4,and the serum IL-6 concentrations were increased at T3 in group DS (P＜0.05).Conclusion Dexmedetomidine used to supplement analgesia with sufentanil can alleviate the stress response and inflammatory response after cardiac valve replacement with CPB.

Objective To compare the effects of nasogastric tube and spiral nasointestinal tube on patients with severe brain injury. Methods Pa?tients receiving enteral nutrition with spiral nasointestinal tube or nasogastric tube were collected and investigated to evaluate the two schemes of en?teral nutrition from aspects of coma score,nutrition improvement,and catheter complications and so on. Results Detection of levels of total protein and prealbumin were conducted for all patients at 7 and 15 days after intubation. Each index was higher in the spiral nasointestinal tube group than in the nasogastric tube group. The reflux and aspiration rate was lower in the spiral nasointestinal tube group than in the nasogastric tube group. The dif?ferences were significant(P<0.05). Conclusion Using spiral nasointestinal tube to give enteral nutrition in patients with severe brain injury can improve the nutritional status,reduce complications,which is more contributory to the recovery.

Objective To investigate the effect of ethanol on level of the main hippocampal subtype of muscarinic receptor(M1)in mice,and evalu?ate whether the content change on this receptor could be linked with alterations in cognition,so as to further reveal the mechanism of brain damage in?duced by ethanol. Methods Sixty female mice were randomly divided into four groups. The model mice were induced by intragastric administration of ethanol at dose of 8%,16%,and 32%respectively of 0.2 mL/10 g for 8 weeks according to the protocol,and control group were treated with intra?gastric administration of distilled water. The capability of learning and memory were examined by Morris water maze,and ELISA method was used to measure the M1 receptor content in hippocampus in each group of mice. Results Compared with first day,the mean escape latency period on the fifth day was significantly shortened in each group. There was no significant difference between ethanol and control group for the mean escape latency period on the fifth day. Compared with the control group,the active time in the target quadrant was significantly shortened in 16%and 32%ethanol group. M1 receptor content in hippocampus formation was significantly decreased in all the ethanol group mice. The ethanol concentration was nega?tive correlated with the M1 receptor content. Conclusion Chronic alcoholism can induce the memory impairment in mice,which might be associat?ed with the low level of M1 receptor subtype in hippocampus of mice.

BACKGROUND: Lentivirus-mediated RNA interference technology has been widely used in the study of gene function because of its specificity, high inhibition efficiency and persistent effects. Virus packaging, transfection, as well as small hairpin RNA (shRNA)-coding sequence could affect the inhibition efficiency. Therefore, impact factors of RNA interference for human Bax inhibitor 1 (BI-1) was studied in this experiment.OBJECTIVE: To find the valid small interference RNA (siRNA) targeting human BI-1 gene by Lentivirus.DESIGN, TIME AND SETTING: Single sample observation was completed in Department of Medical Genetics, Basic Medical College, Peking University Health Science Center from September 2007 to December 2008.MATERIALS: 293T cell and SH-SY5Y cells were preserved in this laboratory; 4 shRNAs were designed through a use of online design tool by Ambion (www.ambion.com). METHODS: Several recombinant plasmids which expressed shRNAs targeting different regions of the BI-1 gene and labeled enhanced green fluorescent protein as a fusion gene were constructed. Four types of shRNA-expressing virions were obtained by cotransfection of shRNA-expressing plasmid and package protein-expressing plasmids into 293T cells. Green fluorescent protein expression was determined using flow cytometry to search the optimal package conditions. Real-time PCR and beta actin mRNA was used as an internal control, four types of recombinant viral supernatant and control viral supernatant were added on the SH-SY5Y cells, knock-down efficacies of the endogenous BI-1 gene were determined, and the RNA interference effective sequences were optimized. MAIN OUTCOME MEASURES: Intracellular green fluorescent protein expression rate of the cells infected with different packaging virus. RESULTS: The optimal infective packaging virus could be obtained when the ratio of plasmids pLentiLox3.7, REV, VSVG and RRE was 2:1:1:1 and the supernatants were collected 48 hours after transfection. Replacement of culture media 24 hours after transfection could increase the infection efficacy of the virions. shRNA targeting-2-17nt (start coding region) of BI-1 gene was most valid interference efficiency and could decrease the gene expression to 40%. CONCLUSION: Influencing factors of Lentivirus-mediated RNA interference technology virus packages were investigated in this study, which could be a foundation for constructing stable BI-1 knocked down neuronal cell model and for studying the abnormal expression of BI-1 related to the neuron apoptosis disease.

OBJECTIVETo explore the in vitro anti-tumor effect and mechanism of dendritic cell (DC) tumor vaccine induced by astragalus polysacharin (APS).

METHODSPeripheral blood mononuclear cells (PBMCs) isolated from human peripheral blood. DCs obtained from human peripheral blood were cultivated and added with culture solution for in vitro inducing them to immature DCs. On the 5th day of culture, 100 microg/mL (as the final concentration) APS was added to cells in the APS group. DCs were induced to mature in the cytokine groups by adding 20 ng/mL rhTNF-alpha (as the final concentration). Changes of morphology and phenotype of DCs were observed. Mature DCs were sensitized with tumor antigen SGC-7901 and co-cultured with allogeneic T cells. The proliferative function of T lymphocytes was detected by MTT assay. Levels of IL-12 and IFN-gamma in co-cultured supernatant were detected by ELISA. Cytotoxic lymphocytes (CTL) activated by DC were co-cultured with tumor cell SGC-7901. The specific killing capacity of CTL to target cells was detected by LDH release assay.

RESULTSThe morphological observation and phenotypic identification of APS induced DCs were in accordance with the characteristics of mature DCs. APS induced mature DCs could stimulate the proliferation of allogeneic T lymphocytes. The proliferation index of T cells increased with increased ratio of stimulator cells to effector cells (P < 0.05). Levels of IL-12 and IFN-gamma in co-culture supernatant significantly increased in a time-dependent manner (P < 0.05). CTL cells activated by sensitization of DCs could significantly kill tumor cells, and the killing effect increased along with increased effector-to-target ratio.

CONCLUSIONAPS could in vitro induce DCs to mature, promote its antigen-presenting capacity, effectively activate CTLs, and enhance anti-tumor function of the organism.

Antigen-Presenting Cells ; cytology ; drug effects ; immunology ; Cancer Vaccines ; immunology ; Cell Line ; Cell Proliferation ; drug effects ; Coculture Techniques ; Dendritic Cells ; cytology ; drug effects ; immunology ; Drugs, Chinese Herbal ; pharmacology ; Humans ; Interferon-gamma ; immunology ; Interleukin-12 ; immunology ; Leukocytes, Mononuclear ; cytology ; immunology ; Lymphocyte Activation ; T-Lymphocytes, Cytotoxic ; cytology ; drug effects

OBJECTIVETo investigate the impact of ethanol extracts from Sedum sarmentosum (ESB) on STAT-3 signaling and its probable molecular mechanism in inducing apoptosis.

METHODMTT assay was used to detect the impact of ESB on HepG2 cell proliferation. FITC-Annexin V-FITC /PI double-labeling were used to investigate the impact on hepatoma carcinoma cell apoptosis. Western blot analysis was used to test the expression levels of cell apoptosis-related proteins Caspase-3, Caspase-9, PARP, P-STAT-3 (Tyr705) , STAT-3, Bcl-2, Mcl-1.

RESULTESB could notably inhibit proliferation of HepG2 cells, and induce HepG2 cell apoptosis, with the dose-dependent inhibitory effect. In addition, ESB could inhibit STAT-3 signaling, down-regulate Mcl-1 and Bcl-2 expressions, and induce degradation/activation of apoptosis-related proteins Caspase-3 and Caspase-9 and PARP degradation in a dose-dependent manner.

CONCLUSIONESB inhibits HepG2 cell proliferation and induces apoptosis by inhibiting STAT-3 signaling and Mcl-1 and Bcl-2 expressions.

Apoptosis ; drug effects ; Blotting, Western ; Caspase 3 ; metabolism ; Caspase 9 ; metabolism ; Cell Proliferation ; drug effects ; Dose-Response Relationship, Drug ; Ethanol ; chemistry ; Flow Cytometry ; Hep G2 Cells ; Humans ; Myeloid Cell Leukemia Sequence 1 Protein ; metabolism ; Plant Extracts ; chemistry ; pharmacology ; Poly(ADP-ribose) Polymerases ; metabolism ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; STAT3 Transcription Factor ; metabolism ; Sedum ; chemistry ; Signal Transduction ; drug effects ; Time Factors

Objective To explore the feasibility of labeling and tracing in vitro goat bone marrow mesenchymal stem cells (BMSCs) by bromodeoxyuridine (BrdU) on the basis of investigation of its optimal concentration, incubating time and cytotoxicity. Methods A healthy goat, aged 10 months old, male, weighing 32 kg, was used in this study. Bone marrow was aspirated. BMSCs were isolated and cultured using the adherence method in vitro. The fourth passage of BMSCs (P4) were incubated with BrdU at 5, 10, 15, 20 μmol/L as 5, 10, 15, 20 μmol/L BMSC groups. Cells were not labeled by BrdU as negative control. The following parameters were measured: induction, differentiation and determination of goat BMSCs; the optimal mass concentration and incubation time of 5-BrdU labeling; cell positive rate at 12, 24, 48 and 72 hours in each group using immunofluoreseenee; the cell survival rate after various concentrations of BrdU ladling by trypan-blue exclusion. Results The morphology of the primary and passage goat BMSCs was fusiform in shape. Goat BMSCs could differentiate into osteoblasts and chondrocytes following induction. BMSC nucleus showed green fluorescence under fluorescence microscope after being labeled by BrdU. The mean labeling rate increased with the increase in the concentration and incubation time of BrdU, and reached to (93.32± 3.25)% after incubation in 15 μmol/L, BrdU for 48 hours. There were no significant differences between 15 μmol/L BrdU for 72 hours, 20 μmol/L BrdU for 48 hours and 72 hours (P > 0.05), or between the other groups or time points (P < 0.05). The labeling rate of the blank control group was 0. The cell survival rate was all above 90% (P > 0.05). Conclusions BrdU can be used as a labeling marker for goat BMSCs. When the concentration is 15 μmol/L and the incubation time is 48 hours, the optimal labeling effect can be achieved. Goat BMSCs labeled with BrdU is of high efficiency and safety.

Objective To explore the best ratio of platelet-rich plasma (PRP) to activator and the synergistic action of thrombin and the growth factors in PRP gel on proliferation of human marrow-derived mesenchymal stem cells (hBMSCs). Methods The activator was made up of 1000 U bovine thrombin and 1 mL 10% calcium chloride solution. PRP was mixed with the activator at the ratios of 1:1, 5:1, 10:1, 20:1, 40:1 respectively in groups A, B, C, D and E. A quantitative sandwich enzyme-linked immunosorbent assay (ELISA) was used for examining the amounts of transforming growth factor-β1 (TGF-β1) and platelet derived growth factnr-AB (PDGF-AB) in PRP gel in each group after incubation for 0,1,8,24, and 120 hours. MTT assay was used to evaluate the effect of PRP gel in each group on hBMSCs proliferation. Results When PRP was mixed with thrombin, concentrations of PDGF-AB and TGF-β1 in PRP gel increased immediately and reached the peak value in 1 hour. The PRP gel in groups B, C, and D contained the highest amounts of PDGF-AB and TGF-β1, and accelerated hBMSCs growth re-markably. The cells proliferation in group A was inhibited. Conclusions The effect of PRP on the hBMSCs proliferation depends on the concentrations of PDGF-AB and TGF-β1 in PRP. Thrombin may influ-ence the hBMSCs proliferation by regulating concentrations of growth factors in PRP to certain extent, but too high a level of thrombin may inhibit hBMSCs growth.

Objective To explore the feasibility of labeling in vitro rabbit osteoblasts with Qtracker and the features of Qtracker-labeled rabbit osteoblasts. Methods A healthy male rabbit, 3 months old, weighing 2 kg, was used in this study. After bone marrow was aspirated, bone marrow stromal cells (BMSCs) were isolated and cultured using the adherence method in vitro. The third passage of BMSCs was induced into osteablasts before incubation with Qtracker at concentrations of 1, 2, 4, 8, 16, 32 nmol/10~6 cells (Groups A, B, C, D, E, F respectively). Cells not labeled by Qtracker served as negative control (Group G). The following parameters were measured: induction, differentiation and determination of rabbit osteoblasts; the optimal mass concentration of Qtracker labeling by fluorescence microscopy and flow cytometry; the cell sur-vival rates at various concentrations of Qtraeker labeling by trypan-blue exclusion; Qtracker-labeled cell pro-liferation by MTr. Results The primary and the passage rabbit BMSCs were chiefly of fusiform shape. Rabbit BMSCs differentiated into osteoblasts following induction. The osteoblasts cytoplasm showed green fluorescence under fluorescence microscopy after being labeled by Qtracker. The mean labeling rate increased with the increased concentration of Qtracker, reaching up to (93.58±2.08) % after incubation at 8 nmol/ 10~6 cells by fluorescence microscopy, and (95.24±1.31) % by flow cytometry. There were no significant differences between Groups D, E, F(P>0.05), but significant differences were found between Groups A, B, C and Groups D, E, F (P<0.05). The labeling rate for Group G was 0. The cell survival rates were all above 96% (P>0.05) . No significant differences were found in the cell proliferation among various con-centrations (P>0.05). Conclusions Qtraeker can be used as a labeling marker for rabbit osteoblasts. When the concentration is at 8 nmol/10~6 cells, optimal labeling effect can be achieved. Rabbit osteoblasts labeled with Qtracker are of high efficiency and safety.