ObjectiveTo evaluate the effects on serum homocysteine(Hcy) level,vascular function and carotid intima-media thickness(IMT) in metformin-treated diabetic patients with or without supplementation with folate and Vitamin B12.MethodsA total of 100 newly diagnosed diabetic type 2patients were divided into two groups by random digits table with 50 cases each,90 patients completed study.Forty-seven participants (control group) received a 6-month course of metformin treatment,43 patients (treatment group) received mefformin,folate and Vitamin B12 treatment.The levels of serum Hcy,endothelin-1 (ET-1),carotid IMT,large or small arterial elasticity index (C1,C2),flow-mediated dilatation (FMD) of the brachial artery were evaluated before and after treatment.ResultsThe level of serum Hcy in control group significantly increased compared with before treatment[ (13.4 ± 2.7)μ mol/L vs.(11.1 ± 1.9)μ mol/L],hut the level of serum Hcy in treatment group significantly decreased compared with before treatment [ (9.2 ± 1.8 ) μ mol/L vs. ( 11.3 ± 2.0) μ mol/L ],there was significant difference(P ＜ 0.05 ).A beneficial effect was observed in the serum ET-1,FMD,carotid IMT and C2 in treatment group[ (20.0 ± 6.2)ng/L,( 15.8 ± 7.6)％,(0.8 ± 0.2) mm,(4.1 ± 2.1 ) ml/mm Hg ( 1 mm Hg =0.133 kPa) × 100 vs. (31.3 ±10.1 ) ng/L,(9.7 ± 4.5)％,( 1.1 ± 0.4) mm,(2.3 ± 1.0) ml/mm Hg × 100 ] (P ＜ 0.05).The levels of ET-1,FMD,carotid IMT and C2 after treatment in control group [ (24.8 ± 6.8) ng/L,( 12.9 ± 6.3 )％,(0.9 ± 0.3)mm,(3.0 ± 1.4) ml/mm Hg × 100] had significant difference compared with before treatment [ (30.6 ± 8.7)ng/L,(9.8 ± 4.6)％,( 1.0 ± 0.3) mm,(2.2 ± 0.9) ml/mm Hg × 100](P＜ 0.05).However,the results were improved significantly in treatment group than those in control group (P ＜0.05).In treatment group,significant correlation were detected between changes of Hcy and ET- 1 (r =0.43,P ＜ 0.05 ),carotid IMT(r =0.56,P ＜ 0.05),FMD (r =-0.54,P ＜ 0.05 ),C2 (r =-0.37,P ＜ 0.05 ).ConclusionsAdministration of folate and Vitamin B12 can reduce the levels of serum Hcy and ET-1 in metformin-treated type 2 diabetic patients,which exert beneficial effect on carotid IMT,FMD and C2.

Calcinosis ; complications ; Decalcification Technique ; Humans ; Meningeal Neoplasms ; complications ; Meningioma ; complications ; Specimen Handling ; methods ; Ultrasonics

In this paper, miRNAs features were briefly introduced and agreeable points were discussed from 4 aspects: organs relationship, syndrome research, Chinese medical pathogeneses, and actions of Chinese herbs. miRNAs, as information media for organs interrelation, was believed to explain Chinese medical pathogeneses and reveal partial molecular mechanisms of Chinese medicine. miRNAs in the body fluid could be taken as one of biological bases of syndromes.
Biomedical Research ; China ; Humans ; Medicine, Chinese Traditional ; trends ; MicroRNAs ; Syndrome

Objective To investigate the efficacy of spectral entropy measurement in reflection of depth of anesthesia and noxious stimulation. Methods Forty-five patients of ASAⅠorⅡ were randomly divided into three groups(n=15).Group A,B and C received fentanyl 1,3 and 5 ?g/kg,respectively,3 min before target controlled infusion(TCI) of propofol.Intubation was performed when the effect-site concentration(CE)reached 3.5 ?g/mL,which was maintained until 5 min after incision.Response entropy(RE),State entropy(SE) as well as heart rate(HR),mean arterial pressure(MAP) were measured at the time points of before fentanyl and 2,3 min after fentanyl,every CE of propofol steps,before intubation,immediately and 1,3,5 min after intubation,before skin incision,and 0.5,1,3,5 min after skin incision,respectively. Results Three minutes after receiving fentanyl,the values of RE and SE in the three groups decreased significantly in a dose-dependent manner,and increased obviously at the same degree during intubation and after skin incision.The values recovered to the level before stress stimulation 1 min after intubation and 5 min after skin incision.There were no differences in the fluctuation of RE and SE among the three groups when the CE of propofol reached 1.0 ?g/mL.Conclusion Spectral entropy may effectively reflect the depth of anesthesia,but not analgesia during anesthesia.

Objective To evaluate the effect of TGF-β3 on rabbit nucleus pulposus( NP) cells cultured in three-dimensional polylactic-co-glycolic acid (PLGA)scaffold in vitro.Methods PLGA scaffolds were fabricated by particulate leaching method and soaked in rabbit NP cells suspension(1 × 106/scaffold).PLGA-seeded NP cells were devided into 4 groups: 100 ng/mL TGF-β3/PLGA,500 ng/mL TGF-β3/PLGA,1 μg/mL TGF-β3/PLGA, PLGA control group.Cell proliferation activity was measured using MTT assay.The glycosaminoglycan ( GAG ) analysis were performed by 1, 9-dimethylmethylene blue(DMMB) assay.mRNA expression was measured by quantitive PCR at each time point.Histological observation was performed to elucidate the morphological changes of NP cells in PLGA effected by TGF-β3.Results Higher cellular proliferation activity, GAG production,Collagen type II, Aggrean expression were observed in TGF-β3 /PLGA-seeded NP cells compared with PLGA control group on day-7,day-14,day-21(P<0.05). Higher dose of TGF-β3 exhibited intense cellular proliferation activity and peri-cellular matrix by increasing trend(P<0.05).Histological observation showed TGF-β3/PLGA developed more significant disc cells cluster than PLGA groups on day-21.Conclusion The 3D porous PLGA scaffold-seeded cells using TGF-β3 can promotes cell proliferation, and prompt extracellular matrix(ECM) production.It is a potential biotherapy for the treatment of disc degeneration.

In order to construct recombinant baculovirus carrying Schistosoma japonicum 26 ku glutathione S-transferase gene (Sj26), and observe the expression of Sj26 in mammalian cells, the Sj26 gene was amplified with plasmid pGEX-3X as template by PCR, and then recombined into T vector for sequencing. Sj26 gene was inserted into the downstream of CMV promoter of donor plasmid pFBDGC, and the recombinant donor plasmid pFBDGC-Sj26 transformed into DH10Bac, then the recombinant bacmid AcCMVSj26 was isolated and transfected into Sf9 cells. The recombinant baculovirus was harvested and final titer of vAcCMVSj26 was measured. BHK cells were transducted with recombinant baculovirus in vitro. By using Western blot, the expression of 26 ku glutathione S-transferase (GST) was detected. The results showed that after enzyme digestion and sequencing, the donor plasmid was successfully constructed. PCR confirmed that pFBDGC-Sj26 and Bacmid homologous recombination occurred in E. coli. After transfection of Sf9 cells with recombinant Bacmid, recombinant baculovirus was replicated in Sf9 cells and expressed green fluorescent protein. PCR further revealed recombinant baculovirus contained Sj26. The titer of the harvested baculovirus was 1.24 x 10(8). Western blot demonstrated that recombinant baculovirus could express 26 ku GST in BHK cells. It was concluded that Sj26 recombinant baculovirus was successfully constructed, and the 26 ku GST was expressed in mammalian cells.

In this study, we designed and synthesized 12 novel aloperine derivatives with different core structures. Among them, compound 3 with a ten-membered ring core was obtained through a special ring expansion reaction after γ-H Huffman elimination of quaternary ammonium salt, and the structure was verified by X-single crystal diffraction. Furthermore, their antiviral activity against human β-coronavirus HCoV-OC43 was evaluated by CCK-8 assay. Quaternary ammonium salt 2a and 3 had a good inhibitory effect against HCoV-OC43, and 2a had the highest anti-HCoV-OC43 activity with an EC₅₀ values of 3.77 μmol·L^-1 and a SI value of over 53.1. Schrӧdinger molecular docking results showed that both 2a and 3 might display their anti-HCoV-OC43 activity by directly acting on host TMPRSS2 and SR-B1. The results expanded the structural types of endocyclic aloperine and the function against coronavirus, and provided useful scientific data for the development of pharmaceutical applications of these compounds.

Objective To construct a prokaryotic plasmid expressing the recombinant protein of enterohemorrhagic Escherichia coli(EHEC) effector NleB1 and to prepare the polyclonal antibody of mouse anti-NleB1.Methods The nleB1 (990 bp) gene was amplified from the genome EHEC O157∶H7 and cloned into the expression plasmid pET24a to construct the recombinant plasmid pET24a-nleB1 that was transformed into E.coli BL21(DE3).After induction with isopropylthio-gelactoside( IPTG) , the His-tag fusion proteins were purified by Ni+affinity chromatography and gel slices.The polyclonal antibody was prepared by immunizing BALB/c mice with purified recombinant proteins and analyzed by Western blotting and ELISA.Results The pET24a-nleB1 recombinant plasmid was successfully constructed, the fusion protein was ex-pressed and purified,and the polyclonal antibody was obtained by immunizing mice with purified fusion protein.Western blotting and ELISA staining demonstrated that the polyclonal antibody was successfully obtained.Conclusion The prepara-tion of the polyclonal antibody against EHEC O157∶H7 NleB1 will be of help for further studies on the function of NleB1 protein.

BACKGROUND: Although previous studies have indicated that transplantation of marrow stromal cells (MSCs) has enhanced axonal regeneration and improved motor dysfunction induced by spinal cord injury. However, it is still unclear how transplanted MSCs promoted axonal regeneration and the relationship of transplanted cells and regenerated axons.OBJECTIVE: By immunofluorescence and immunoelectron microscopy, this study sought to elucidate the mechanism of promoting axonal sprouting following transplantation of MSCs into a completely transected spinal cord.DESIGN, TIME AND SETTING: The in vivo cytology randomized controlled animal experiment was performed at the Department of Anatomy of National University of Singapore from March 2006 to June 2007.MATERIALS: MSCs were isolated and purified from a Wistar neonatal rat. Model of completely transected spinal cord injury was established by transection at T_(10) segment with asepsis technique using 36 clean adult female Wistar rats.METHODS: MSCs were subcultured and purified. Thirty-six adult female Wistar rats were randomly divided into transplanted and control groups, with 18 animals in each group. Following 9 days of completely transected spinal cord injury, rats in the transplanted group were injected with MSCs (1×10~(11)/L), 5 μL in the defect region and 2.5 μL in 1 mm upper and lower the defect region. Rats in the control group were infused with an equal volume of DMEM, at the speed of 1 μL/min.MAIN OUTCOME MESSURES: Survival and differentiation of transplanted MSCs; Regeneration of axon and survival of host-derived nestin-, NF200-, GFAP-, and CNP-positive cells in control and transplanted groups; Relationship of regenerated axon and CNP-positive cells RESULTS: Two weeks after transplantation, a large number of CFDA-SE labeled MSCs were detected in the lesion site. Survival transplanted cell number was decreased over time. Abundant 0×42-positive phagocytes/activated microglia and cavity might affect the survival of transplanted cells. The number of MSCs was decreased, but MSC transplantation could promote axonal regeneration in the lesion site, and enhance the survival of host-derived nestin-, NF200-, GFAP-, and CNP-positive cells in lesion site, and host-derived CNP-positive cells and Schwann cells provided structural support to regenerated axons and promote their remyelination in spinal cord injury.CONCLUSION: Transplantation of MSCs enhances survival of host-derived CNP- positive cells and Schwann cells, and which may provide structural support to regenerated axons and promote their remyelination in spinal cord injury.

Animals ; Arecoline ; pharmacology ; Cholinergic Agonists ; pharmacology ; Female ; Guinea Pigs ; In Vitro Techniques ; Male ; Muscle Contraction ; drug effects ; Muscle, Smooth ; physiology ; Pyloric Antrum ; physiology ; Receptor, Muscarinic M3 ; metabolism