Objective To evaluate the adhesion, proliferation and osteogenic differentiation of rabbits' mesenchymal stem cells (MSCs) cultured in demineralized bone matrix coated with OB-Cadherin. Method The second generation of MSC s were seeded onto the OB-Cadherin cover over allogenic frozen-dried demineralized bone matrix(FDBM) and the FDBM without OB-Cadhefin as control, and then both were cultured separably in vitro. The densities of seeded cells, the adhesion rate were measured, and their ALP activity was assayed in order to take it as an index of cell adhesion, proliferation and osteogenic differentiation. The growth and adhension of MSCs on the FDBM was observed and evaluated microscopically and electronic scanning microscopically. Data were ex-pressed as means and standard deviation (x±s), and were analyzed with SPSS 12.0. Independent-Sanples T-test and Paired t test was used, and P<0.05 indicated statistically significant. Results There was no significant dif-ference in cell proliferation between modified FDBM and unmodified FDBM cultured fot 7, 14, 21 d. ays. After culture for 20 hours, the adhesion rate in the control group was (35.56±1.75)%, the densities of seeded cells were less than 2.7×104. The adhesion rate of cells in the modified group was consistent at 80%, whereas the densities of seeded cells were as high as 5.0×105 compared with control group (P<0.01). After cultured for 20 hours, the efficiency of cell adhesion in the modified FDBM was higher than that in the unmodified FDBM. After cultured for 7 days, the cultured MSC on modified FDBM expressed higher AIP activity, and after cultured for 14 days, the ALP activity on modified FDBM was much higher than that on unmodified FDBM (p<0.01). After cultured for 14 days, osteocalcin immunohistochemical positive rate of modified group was (71±11)%, while that of the control group was(49±8)% with significant difference Conclusion OB-Cadherin enhances cell adhesion to FDBM and promotes MSC to differentiate to osteoblast, but no obvious effects d OB-Cadherin on cell proliferation were observed.

Objective:To investigate the specific immune act ivated effects in the mouse immunized by MHC I-restriced tumor antigen peptide pulsed dendritic cells(DC) with IL-2 gene modified.Methods:DC were transfected with IL-2 gene via adenovirus vector(DC-IL-2) the mRNA of mIL-2 was examined by RT-PCR.Syngeneic mice were immunized by DC-IL-2 pul sed with MHC-I-restricted Mutl tumor antigen peptiede of 3LL Lewis lung carcin oma (DC-IL-2-Mutl),the effects of lymphocyte subsets in the draining lymphoid node from the tumor antigen pulsed DCs with gene modified immunized mice by flo w-cytometric analysis(FACS) analysis.Results:The mRNA of mIL-2 was expressed in the DCs.The ratio of CD8+T cells was hosted in the dra ining lymphoid node from Mutl pulsed DCs(DC-Mutl) immunized mice.The ratio of C D8+T cells and NK cells were all hosted obviously in the draining lymphoid nod e from DC-IL-2-Mutl immunized mice .Conclusion:Immunized mice with DC-IL-2-Mutl can induce the mouse specific antitumor immunized affect in vivo and can activate a variety of immunized effect in the mouse.

Objective To study the inlfuence factors on detection of activity of four coagulation factors in prothrombin complex concentrates (PCC) by several factors. Methods Using Pharmacopoeia of the People’s Republic of China (2010) as reference, the activity of four coagulation factors in PCC were investigated by choosing different pre-treatments, different diluents, different salt concentration, different standard human plasma and different company reagents. Results The activity of FII, FVII, FX were decreased and FIX was increased in the condition of adding protamine sulfate, and there were no differences of four coagulation factors whether warm bath in 37 for 15 min or not. However, the differences of four coagulation factors were significant by using deficient plasma, saline, distilled water and commercial dilution buffer(P<0.05). The activity of coagulation factor II, X in 1 mol/L salt concentration of PCC were significantly lower than in 0.25 mol/L(P<0.05), while coagulation factor VII, IX were not. The activity of FII, FVII, FIX, and FX were different by using different standard human plasma to make standard curve. The activity of four coagulation factors existed significant difference(P=0.00) by using SIEMENS company reagents and domestic reagents. Conclusion Choosing different pre-treatments, different dilution buffers, salt concentration, standard human plasma and commercial kits will inlfuence the detection result of coagulation factors.

AIM:To analyze the expression profile of long non-coding RNA (lncRNA) in the liver tissues of drug-induced liver injury ( DILI) and immune liver injury ( ILI) .METHODS:The technique of lncRNA microarray was used to inspect the lncRNA expression profile in the mouse liver tissues that the liver injury was induced by acetaminophen or concanavalin A .The raw data of lncRNA were pretreated for normalization .RESULTS:Compared with normal hepatic tissue, the lncRNA which had more than 1.5-fold variation and significant difference (P<0.05) by statistical analysis were regarded as lncRNA with differential expression .A total of 68 lncRNA with differential expression were found in the hepatic tissues of DILI, with 21 increased more than 1.5 folds and 47 reduced more than 1.5 folds.A total of 60 lncRNA with differential expression in the liver tissues of ILI were observed , with 17 increased more than 1.5 folds and 43 reduced more than 1.5 folds.In all lncRNA, 8 was simultaneously up-regulated in 2 liver injury models , accounting for 38%and 47%respectively, while 28 was simultaneously down-regulated in 2 liver injury models, accounting for 59%and 65%re-spectively .CONCLUSION:lncRNA expression profiles of DILI and ILI change significantly in comparison with normal hepatic tissue , and there are also differences between 2 hepatic damage models .The simultaneous changes of lncRNA may participate in the same or similar pathophysiological process , while the differences may be involved in relatively particular mechanisms .

OBJECTIVE:To observe clinical efficacy and safety of naloxone hydrochloride combined with Danshen injection in the treatment of neonatal hypoxic ischemic encephalopathy(HIE). METHODS:104 patients with HIE were randomly divided into observation group and control group with 52 cases in each group. The control group was treated with Naloxone hydrochloride injec-tion 0.02 mg/kg,ivgtt,qd and conventional symptomatic treatment;while observation group was additionally treated with Danshen injection 4-6 ml+10% Glucose injection 20 ml,ivgtt,qd,on the basis of the control group. Treatment course of 2 groups lasted for 7 d. Clinical efficacy,symptom and sign recovery of 2 groups were compared,as well as levels of serum MMP-9,IL-6 and TNF-α,cerebral hemodynamic parameters and ADR. RESULTS:There was no statistical significance in MMP-9,IL-6,TNF-α and cerebral hemodynamic parameters before treatment(P>0.05). The total effective rate of observation group(92.31%)was signifi-cantly higher than that of control group (75.00%);after treatment,the recovery time of consciousness disturbance,primitive re-flex and muscle tension in the observation group were shorter than those in the control group;the levels of serum MMP-9,IL-6 and TNF-α in observation group were significantly lower than those in control group;peak systolic flow velocity(PSFV),end-dia-stolic flow velocity (EDTV) were higher in the observation group than in the control group,with statistical significance (P<0.05);resistance index (RI) was lower than in control group,but without statistical significance (P>0.05). No ADR was ob-served in 2 groups. CONCLUSIONS:Naloxone hydrochloride combined with Danshen injection can significantly promote HIE and signs recovery,reduce the levels of serum MMP-9,IL-6 and TNF-α,and improve hemodynamic parameters with good safety.

BACKGROUND:Transplanted neural stem cel s can survive, proliferate and differentiate into neurons and/or glial cel s in the host, thereby promoting partial function recovery in the host.
OBJECTIVE:To study the therapeutic effects of neural stem cel transplantation on cerebral palsy rats. METHODS:Twenty-four rats were randomly divided into three groups:control group, model group and transplantation group. Animal models of cerebral palsy were made in the latter two groups. One week after modeling, rats in the transplantation group were injected 1 mL stem cel suspension (1×105) via the jugular vein, and rats in the control and model group were given the same volume of normal saline. Toe distance, step length and elevated body swing test in rats were detected, and histopathological changes in the rat brain were observed 3 weeks after transplantation.
RESULTS AND CONCLUSION:In the model group, the toe distance and step length of the front left palm were significantly lower than those of the front right palm (P<0.05). Compared with the model group, the toe distance and step length in the transplantation and control groups were significantly increased (P<0.05). In the elevated body swing test, rats in the model group presented with asymmetric swing of the
body, but rats in the other two groups exhibited symmetric swing of the body (P<0.05). Additional y, the ratio of right to left hemispheric areas was significantly higher in the transplantation and control groups compared with the model group (P<0.05). In conclusion, neural stem cel transplantation via the jugular vein can improve brain function and restore motor function in rats with cerebral palsy.

Objective To observe the effect of compound fetal liver extract combined with antiviral therapy on liver fibrosis in patients with cirrhosis.Methods 60 patients with liver cirrhosis from April 2014 to July 2015 were randomly divided into treatment group and control group(n=30).The patients in control group were treated with conventional anti viral therapy , 30 cases in treatment group were treated with compound fetal liver extract combined with antiviral therapy postoperative.Results 1 month after treatment, the treatment group serum liver fibrosis HA, PCM value respectively(107.5 ±17.8,99.8 ±14.9)ng/mL, were lower than in the control group (138.4 ±15.2,124.1 ±18.1)ng/mL(P<0.05).1 month after treatment, the treatment group liver fibrosis collagen type IV, III procollagen value respectively(58.9 ±11.0,109.2 ±11.1)μg/L, were lower than in the control group (85.7 ±11.2,122.7 ±11.3)μg/L(P<0.05).Conclusion Compound fetal bovine liver extract combined with anti-viral therapy in patients with cirrhosis has good, better than the use of antiviral drugs alone.

Objective To study the influence of prothrombin complex concentrates (PCC) containing different levels of antithrombin (AT) and heparin on its procoagulation and anticoagulation activities.Methods PCC containing traces of heparin and AT were prepared.The design of orthogonal experiment: set AT and heparin two variable factors, each factor respectively from three levels: 0, 0.4, 0.8 IU/mL; 0, 2.5, 5.0 IU/mL.Their influence on the procoagulation and anticoagulation activities of PCC were analysed.Results The R value of the influence of AT on thrombin inhibitory capacities was higher than that of heparin.With the increase of AT level, time to peak gradually extended.Lag time of 0.4 IU/mL and 0.8 IU/mL AT was the same.The peak height of 0.4 IU/mL AT was highest.The higher the level of AT, the greater the impact on thrombin inhibitory capacities.0.4 IU/mL and 0.8 IU/mL AT had similar impact on thrombin inhibitory capacities.The R values of the influence of heparin on thrombin generation were higher than that of AT.With the increase of heparin level, lag time and time to peak were longer while the peak height declined sharply.0 IU/mL and 2.5 IU/mL heparin had similar influence on thrombin generation.The higher the level of heparin, the greater influence on thrombin inhibitory capacities.2.5 IU/mL and 5.0 IU/mL heparin had similar influence on thrombin inhibitory capacities.Conclusion AT has greater influence on the anticoagulation activities of PCC than that of heparin, while heparin has greater influence on the procoagulation activities of PCC than that of AT.PCC containing medium levels of AT and heparin is relatively balanced between procoagulation and anticoagulation.

BACKGROUND:Transplantation of mesenchymal stem cels to prevent and treat degeneration of the intervertebral disc is a feasible method. Mesenchymal stem cels co-transfected by SRY-related high mobility group-box gene 9 (SOX-9) and growth differentiation factor-5 (GDF-5) can differentiate into nucleus pulposus cels, in order to obtain greater effect of induction and proliferation of nucleus pulposus cels. OBJECTIVE:To investigate the effect of SOX-9 and GDF-5 co-transfection on the differentiation of rabbit bone marrow mesenchymal stem cels into nucleus pulposus cels. METHODS: We separated and cultured bone marrow mesenchymal stem cels from the bone marrow of rabbit aged 4 months. Passage 3 cels were divided into five groups andin vitro induced to differentiate into nucleus pulposus cels: non-transfected group, empty vector transfection group, SOX-9 transfection group, GDF-5 transfection group, SOX-9 and GDF-5 co-transfection group. At 14 days after transfection, RT-PCR was employed to assay SOX-9, GDF-5 and colagen type II mRNA expressions in bone marrow mesenchymal stem cels. The marker of nucleus pulposus cels-KRT19 expression was also detected by immunohistochemical staining. RESULTS AND CONCLUSION:In the co-transfection group, the mRNA expressions of SOX-9, GDF-5, and colagen type II were significantly higher than those in the SOX-9 transfection group, GDF-5 transfection group, and both these two groups, respectively (P < 0.05). Cels were positive for KRT19 in the SOX-9 and GDF-5 groups, and strongly positive for KRT19 in the co-transfection group. These findings indicate that double gene-transfected bone marrow mesenchymal stem cels are better than single gene-transfected cels with regard to differentiation into nucleus pulposus cels and secretion of extracelular matrix.

Objective To investigate the effects of alpha lipoic acid on patients with diabetic retinopathy (DR),explore the possible mechanisms,and provide experimental evidence for the treatment of DR with lipoic acid.Methods Totally 75 DR patients were randomly divided into control group and treatment group.Patients in control group received the standard hypoglycemic treatment,antihypertensive therapy,and other comprehensive treatment.Patients from the treatment group were treated with alpha lipoic acid 600 mg + 250 ml 0.9％ sodium chloride injection additionally,avoiding light intravenous therapy,for a total of 4 weeks.Before and after treatment,fundus fluorescein angiography and visual inspection were detected in two groups.In addition,serum level of asymmetric dimethylarginine (ADMA) were detected with high performance liquid chromatography,serum lipid peroxidation product malondialdehyde (MDA) content and superoxide dismutase (SOD) activity were detected to reflect the in vivo oxidative stress before and after treatment in two groups.Results Compared to the control group,visual acuity was improved significantly (P ＜ 0.05),and total efficiency of ocular lesions-improved was increased significantly (84.21％ vs 40.54％,x2 =15.28,P ＜ 0.01).Serum levels of endogenous inhibitor of nitric oxide synthase ADMA [(0.32 ±0.13) μmol/L vs (0.51 ± 0.26) μmol/L,P ＜ 0.05] and MDA [(3.22 ± 0.90) nmol/L vs (5.13 ± 0.42) nmol/ml,P ＜ 0.05] were significantly decreased,while SOD levels [(72.68 ± 1.31) U/ml vs (53.16 ± 1.54) U/ml,P ＜0.05] were significantly increased in DR patients after alpha lipoic acid treatment.Conclusions Alpha lipoic acid had some therapeutic effects for DR patients,which mainly lied in the improvement of visual acuity and eye disease,might be related to the decrease of oxidative stress and serum level of endogenous nitric oxide synthase (NOS) inhibitor ADMA.