Objective To purify recombinant methioninase and investigate the synergetic inhibitory effect of methioninase and cisplatin on the proliferation of human lung adenocarcinoma cell line GLC. Methods Recombinant methioninase was purified with GST-column from supernatant after ultrasonic disruption of cultured Escherichia coli in the prokaryotic expression system pGEX-4T-1-Met/Dh5a. MTT assay was used to determine the inhibition rate of methioninase in combined with cisplatin on cell proliferation,and their synergistic effect was evaluated by using the q value judge method. Results The concentration of recombinant methionine cleaving enzyme was 0.22 mg/mL, the purity was 95%, and the activity was 0.568 IU/mg. After 72 h culturing, the inhibition rate of cisplatin methionine was 24.80%and 27.49%respectively,while the inhibition rate of the combined drugs was 66.80% ( =1.47>1.15) which showed a significant synergistic effect. Conclusion Both methioninase and cisplatin have the inhibition effect,and the combined drugs display a significant synergistic effect on the proliferation of GLC.

Objective To explore the method and efficiency of percutaneous transluminal septal tunnet myocardial ablation(PTSTMA) in treatment of 26 cases hypertrophic obstructive cardiomyopathy (HOCM) who were not suitable for conventional technology.Methods Firstly,we used a monorail Balloon which was slightly bigger than the interventricular septal branch of coronary artery and dilated it until posterior septal.After that,an OTW Balloon with larger size than the monorail was used to dilate again until made aventricular septum tunnel.Then,some alcohol was injected and PTSTMA was performed.Finally,we did the other and/or another interventricular septal branch by above method until the left ventricular outflow tract pressure gradient (LVOTPG) reduced ≥50％.The clinical indexes of the 26 cases HOCM immediately pest-operation of PTSTMA were observed and the follow up data during short term and metaphase were analyzed.Results The LVOTPG reduced ≥50％ in the26 cases HOCM,immediately after PTSTMA,the LVOTPG reduced from (75.6 ±22.4)mm Hg to (21.4 ± 5.8) mm Hg (t =11.94,P ＜ 0.01).At three months after ablation,the thickness of septal myocardium reduced from (22.8 ± 5.8) ram before ablation to (16.8 ± 4.2) mm(t =4.27,P ＜ 0.01),left atrium dimension reduced from(48.0 ±7.0) mm to (42.0 ±8.6) mm (t =2.76,P ＜0.01).Followed up 6.0to 60.0 months,the patients suffering from chest pain reduced from 14 cases before to 4 cases after the procedure(53.8％ (14/26) vs 15.4％ (4/26),x2 =8.49,P ＜ 0.01),the patients with expiratory dyspnea reduced from 26 cases to 5 cases(100％ (26/26) vs 19.2％ (5/26),x2 =35.22,P ＜ 0.01),NYHA functional class improved from (2.4 ± 0.6) to (1.4 ± 0.7) (t =5.53,P ＜ 0.01).Conclusion The PTSTMA was a supplemental method of PTSMA on treating HOCM,which was safe and useful during the short term and metaphase.

Objective To establish the anti-infective tissue engineered bones (TEBs) and evaluate the anti-infective and osteogenic effects of the infection-prevention TEBs on femoral large bone defects in goats.Methods Based on the controlledrelease antibiotic system fibrin gel-coated vancomycin alginate beads (FG-Vanco-AB),the infection-prevention TEBs were established and evaluated.They were transplanted into the critical-size defects in the right femurs of goats.TEBs without the controlled-release antibiotic system were used as controls and transplanted into the left femoral defects.The breakpoint sensitivity of vancomycin (5 mg/mL) for S.aureus was used as a standard concentration.Postoperatively,the vancomycin concentrations in the lesion site,in the adjacent site and in the circulation,as well as the anti-infective effects of the infection-prevention TEBs were evaluated by High-performance liquid chromatography (HPLC).Bone hcaling was assessed by histology,CT and ECT.The results were used to evaluate the osteogenic effect of the infection-prevention TEBs.Results Results from ESM,CLSM and in vivo tracing showed that the in vitro and in vivo survival conditions of seeded cells were analogous to those of TEBs.The effective concentration (over the bactericidal concentration) of vancomycin in bilateral defects and in blood lasted for 28 days,2 days and 7 days,respectively.The concentration of vancomycin in the femur decreased gradually from the grafted site to both ends.At 28 and 56 days postoperatively,the ECT results showed no significant difference between the right and left femurs.CT and histology demonstrated that at 14,28 and 112 days after surgery,bone defects in the bilateral femurs were repaired synchronously,and were completely covered by new bone tissue after 112 days.Conclusion The anti-infective TEBs were successfully established.FG-Vanco -AB in the transplanted sites provided the local bone tissues with anti-infective capability whilst not interfered the process of bone reconstnction and wound healing.

Objective To systematically evaluate the efficacy and safety of amlodipine (A)/hydrochlorothiazide(H) versus val‐sartan(V)/hydrochlorothiazide(H) in treatment of essential hypertension .Methods Literature was retrieved online in Cochrane Li‐brary ,PubMed ,OVID ,MEDLINE ,EMBASE ,CBM ,CNKI ,VIP and Wan fang database up to November 2013 .Relevant magazines were retrieved manually .Quality of the included studies was assessed and Meta‐analysis was performed with RevMan 5 .2 software . Results Seven randomized controlled trials(RCTs) were finally included .Meta‐analyses showed that :in terms of lowering ABP ,V/H group was more effective than A/H group ,the difference was statistically significant (P<0 .05);there was no significant differ‐ence in the decreased value of clinic BP and the control rate of blood pressure between A /H group and V/H group(P>0 .05) .Ad‐verse events occurred less frequently with V/H group compared with A/H group ,the difference was statistically significant (P<0 .05) .Conclusion A/H treatment of essential hypertension is inferior to V/H ,and has more adverse events .

Objective To investigate the mechanism and prevention of syncope on patients with hypertrophic cardiomyopathy (HCM). Methods Seventy-six cases of HOCM (obstruction group) were successfully operated by PTSMA and oral ACEI/ARB. After six months , they were treated with β-receptor blocker. Another 29 patients (control group) with LVOTPG < 50 mmHg or < 70 mmHg after pharmacologic stress test (PST), have being treated with β-receptor blocker. The results was observed as follow: (1) the difference of between syncope incidence and positive incidence induced by PST in control group; (2) the difference of syncope incidence at half year, a year and admission in control group; (3) the difference of syncope incidence in obstruction group before and after operation; (4) the difference of syncope incidence after six months between two groups at same period; (5) the difference of syncope incidence one year between two groups. Results In control group, the syncope positive incidence induced PST was 55.5%. Treating with medications for half a year , syncope incidence significantly dropped than that on admission (P < 0.05); Obstruction group syncope incidence in the history obviously lower than the control group syncope positive induced PST (P < 0.05), and half a year after takingβ-receptor blocker syncope incidence was significantly dropped than before (P < 0.01). Conclusions The mechanism of syncope with HOCM is not only obstruction but also neuronal reflex. PST is an very useful inspection item for screening the ablation indication, analysis syncope mechanism, and guiding clinical medication.β-receptor blocker is an effective drug on treating and preventing syncope with HCM.

Objective To investigate the effect of chronic stress on rat behavior changes,and specific protein neurogranin (Ng) level changes to explore the control efficiency ofginsenoside Rgl in cognitive impairment.Methods Thirty-six adult male SD rats were randomly divided into control group,model group (chronic unpredictable stress animal models,CUS),and CUS-G treatment group.The chronic stress models in the later two groups were established by CUS;rats in the CUS-G treatment group were given 11 00 mg/kg ginsenoside Rgl;Behavior changes of rats were detected by sugar consumption test and body weight measuring.Morris water maze test was used to study the learning and memory abilities.The Ng content in the cortex and hippocampus was detected by Western blotting.The Ng expression in the cortex and hippocampus was measured by immumohistochemical staining.Morphological changes in the target areas of animal models were detected by HE staining.Results As compared with the control group and CUS-G treatment group,the CUS model group had significantly decreased sugar consumption and weight (P＜0.05).The water maze test showed that learning and memory abilities in rats decreased significantly after chronic stress,and the escape latency in the CUS-G treatment group was reduced,which showed significant difference as compared with that in the control group and CUS model group (P＜0.05).As compared with the control group,the CUS model group had significantly decreased Ng content in the cerebral cortex and hippocampus and average absorbance values of Ng (P＜0.05),while the CUS-G treatment group had significantly increased Ng content in the cerebral cortex and hippocampus and average absorbance values of Ng as compared with the CUS model group (P＜0.05).Conclusions Ginsensode Rg1 (100 mg/kg) can increase the level of Ng in the cerebral cortex and hippocampus to restore the damage of cognitive ability.

Objective To investigate the effects of exosomes derived from human dental pulp mes-enchymal stem cells ( hDPSC-exosomes) on the maturation and function of dendritic cells ( DC) stimulated by lipopolysaccharide ( LPS ) , and to evaluate their regulatory effects on the immune system. Methods Adult permanent teeth-derived dental pulp mesenchymal stem cells were cultured in vitro to extract exosomes in the cell culture medium. The morphology and sizes of the exosomes were observed under transmission electron microscopy. Expression of CD9 and CD63 on the surface of the exosomes was detected by Western blot. PBS, LPS and LPS+hDPSC-exosomes were respectively used to stimulate mouse bone marrow-derived dendritic cells (DC2. 4) for 24 h. A blank control group was set up accordingly. Expression of co-stimulato-ry molecules and cytokine secretion were detected by flow cytometry and ELISA, respectively. Expression of TLR2, TLR4 and NF-κB at mRNA level was detected by RT-PCR. Changes in the functions of DC were evaluated by mixed lymphocyte reaction ( MLR) . Results Adult permanent teeth-derived dental pulp mes-enchymal stem cells were successfully isolated. Up-regulated CD73 and CD90, and down-regulated CD45 were detected on the surface of the cells. Under electron microscopy ( SEM ) , hDPSC-exosomes showed round or oval microcapsule bodies about 50-80 nm in diameter with positive surface markers of CD9 and CD63. hDPSCs-exosomes could significantly reduce the LPS-induced expression of co-stimulatory molecules CD11c and CD86 on DC surface. Moreover, hDPSCs-exosomes increased TGF-β expression and decreased IL-4. They could also significantly inhibit the proliferation of splenic lymphocytes that was induced by DC af-ter LPS stimulation. Compared with the blank control group, hDPSC-exosomes could promote the expression of TLR2 and TLR4 on DC surface and up-regulate the expression of NF-κB. Conclusions This study showed that hDPSC-exosomes could inhibit the activation and functional maturation of DC, promote the de-velopment towards tolerant DC through TLR-NF-κB signaling pathway, and induce immune tolerance to regu-late immune balance.

Objective : To investigate the effect of resveratrol (RSV) in the treatment of peri-implantitis in a murine model and its effect on nuclear factor kappa-B (NF-κB) signaling and mitogen-activated protein kinase (MAPKs) signaling. Methods: This study has been reviewed and approved by the Ethics. After extracting the right maxillary molars of 40 C57BL/6 mice and allowing them to heal naturally for 8 weeks, implants were implanted at the site of the first molar. The mice were randomly divided into a control group, a mouse peri implantitis model group, a low-dose group of 20 mg/kg resveratrol (RSV-L), and a high-dose group of 40 mg/kg resveratrol (RSV-H). After 4 weeks of implant implantation, a silk thread ligation induced peri implantitis model was established in all mice except for the control group. The model group received intervention with physiological saline by gavage, while the drug group received intervention with resveratrol by gavage for 6 consecutive weeks. After 6-week treatment, observe the swelling of the gums around the implant and measure the bone resorption around the mouse implant using micro CT. Enzyme linked immunosorbent assay (ELISA) was used to detect the levels of tumor necrosis factor alpha (TNF-α) and interleukin-6 (IL-6) in gingival crevicular fluid. HE staining was used to observe the infiltration of inflammatory cells in the surrounding tissues of mouse implants. Protein expression level and phosphorylation level of extracellular regulated protein kinases (ERK), p-ERK, c-Jun N-terminal kinase (JNK), p-JNK, p38 mitogen activated protein kinase (p38 MAPK), p-p38MAPK, nuclear factor kappa-B (NF-κB), p-NF-κB, nuclear factor-κB inhibitory protein (IκΒα), p-IκBα in MAPKs/NF-κB signaling pathway were detected by Western blot (WB). Results: Resveratrol group showed reduced tissue edema and decreased alveolar bone resorption. Among them, the high-dose resveratrol group had lighter tissue edema and weaker bone resorption compared to the low-dose group. The micro CT results showed that significant changes in the bone level around the implant were observed in the model group mice at four sites: proximal, distal, buccal, and palatal. High dose resveratrol intervention reduced alveolar bone resorption (P<0.05); compared with the low-dose group, the high-dose group showed a decrease in palatal bone resorption (P<0.05), while there was no significant difference in absorption between the mesial, distal, and buccal sides (P>0.05). The ELISA results showed that compared with the model group, the levels of TNF - α and IL-6 in the gingival crevicular fluid of mice in the low-dose and high-dose resveratrol groups were lower (P<0.05). The IL-6 in the gingival crevicular fluid of mice in the high-dose resveratrol group was lower than that in the low-dose group (P<0.05). However, there was no significant difference in TNF-α levels between the two groups. HE staining showed a decrease in inflammatory cell infiltration in mice after treatment with resveratrol. The WB results showed that compared with the control group, the expression levels of p-Erk, p-JNK, p-p38MAPK, p-IκA, and p-NF-κB phosphorylated proteins in the gingival tissue of the model group mice were significantly increased (P<0.01). The resveratrol treatment group significantly inhibited the phosphorylation of p-Erk, p-JNK, p-p38MAPK, p-IκA, and p-NF-κB proteins. Compared with the low-dose group, the high-dose group inhibited the phosphorylation of MAPKs/NF-κB signaling pathway related proteins more significantly (P<0.05). Conclusion Resveratrol protect ligature induced peri-implantitis murine model, which may be related to its inhibition of phosphorylation of MAPKs/NF-κB pathway.