Objective To investigate the brain protective effects of ropivacaine on cerebral ischemia-reperfusion injury in rats and its influence on cyclic adenosine-adenosine synthase/stimulator of interferon gene (cGAS/STING) pathway.Methods SD rats were randomly divided into sham operation group,model group,low-dose ropivacaine group (0.075 mg/kg),medium-dose ropivacaine group(0.15 mg/kg),and high-dose ropivacaine group (0.30 mg/kg),positive control group(nimodipine,5 mg/kg),with 8 rats in each group,except for the sham operation group,the rats in the other groups were given suture method to construct a rat model of cerebral ischemia and reperfusion,and the neurological deficit scores of the rats in each group on the 0th,3rd,and 7th day of administration were measured;hematoxylin-eosin staining (HE) was used to detect the pathological changes of brain tissues in each group;2、3、5-Triphenyte-trazoliumchloride(TTC) method was used to detect cerebral infarction area; enzyme-linked immunosorbent (ELISA) was used to detect the levels of tumor necrosis factor(TNF-α),interleukin(IL-1β),IL-6,superoxide dismutase(SOD) and malondialdehyde (MDA) in brain tissue; qRT-PCR was used to detect the level of mitochondrial DNA (mtDNA);Western blotting was used to detect the expression of cGAS/STING pathway protein in the brain tissues.Results Compared with the sham operation group,the brain tissue of the rats in the model group was seriously damaged,cerebral infarction area,the nerve defect score,TNF-α,IL-1β,IL-6,MDA levels in brain tissue,Cytoplasmic mtDNA level,cGAS and STING proteins expression were significantly increased,and the SOD levels in brain tissue were significantly reduced (P<0.05);compared with those in the model group,the brain tissue damage of the rats in the ropivacaine groups and the positive control group was relieved,cerebral infarction area,the nerve defect score,TNF-α,IL-1β,IL-6,MDA levels in brain tissue,Cytoplasmic mtDNA level,cGAS,STING proteins expression were significantly reduced,and SOD levels in brain tissue were significantly increased (P<0.05). Conclusion Ropivacaine has a certain protective effect on cerebral ischemia-reperfusion injury,and can inhibit cGAS/STING pathway.

Objective To accurately understand multi-spot medical practice in China and evaluate its effect.Methods Operation data of 2008 and 2009 were gathered from all medical institutions in Kunming to analyze the effect of the practice trial using the fixed effect model.Results Under control of such factors as the scale and type of the hospitals,clinic visits of the second certified hospital benefited by the practice increased by 14％than before,whereas the medical revenue of the institution increased by 29％.The effect proves more significant in terms of total clinic visits and medical revenues at level-1 and level-2 medical institutions.Conclusion Multi-spot medical practice in Kunming proves successful as it increased medical service supply and medical revenue,especially for level-1 and level-2 medical institutions.This effect possibly results from releasing high-level physicians down to lower-level institutions,helping these institutions to attract patients.

ObjectiveTo explore whether miR-375 regulates the malignant characteristics of osteosarcoma (OS) by influencing the expression of MMP13. MethodsPlasmid DNAs and miRNAs were transfected into OS cells and HEK293 cells using Lipofectamine 3000 reagent. Real-time quantitative polymerase chain reaction was performed to measure the expression of miR-375 and MMP13 in OS patients and OS cells. Western blot was performed to analyze the MMP13 protein in the patients with OS and OS cells. The targeting relationship between miR-375 and MMP13 was analyzed by luciferase assay. Migration and invasion were analysed by heal wound and transwell assays, respectively. ResultsmiR-375 expression in OS tissues was lower than that in normal tissues. The expression of MMP13 was upregulated in OS tissues. MMP13 expression was negatively correlated withmiR-375 expression in patients with OS. Migration and invasion were significantly inhibited in OS cells with the miR-375 mimic compared with OS cells with the miRNA control. MMP13 partially reversed the inhibition of migration and invasion induced by miR-375 in the OS cells. ConclusionmiR-375 attenuates migration and invasion by downregulating the expression of MMP13 in OS cells.

Objective To clinically observe and verificate the advantages of treating infantile anorexia of spleen-dysfunction in transportation with infantile massage and Chinese medicine based on the thought of “invigo-rating spleen and nourishing qi”. Methods Multi-central large-sample randomized controlled trial was carried out in Guangzhou, Shenzhen, and Foshan. Five hundreds cases of infantile anorexia of spleen-dysfunction in transportation were randomized into infantile massage group, Chinese medicine group, infantile massage combined with Chinese medicine group and Western medicine group and two sessions of treatment were required in each group. Separately, during the enrollment, after 2 sessions of treatment, infantile anorexia signs and symptoms scale , the related therapeutic effect criterion from clinical guideline of new drugs for traditional Chinese medicine, the expression level of hemoglobin and blood zinc were adopted for efficacy assessment. Results A mong 4 groups , the recovery rate and the total effective rate of infantile massage combined with Chinese medicine group were better than those of other groups (P < 0.05). The expression level of hemoglobin and blood zinc of 4 groups both increased obviously (all P < 0.01), but more improvement was found in infantile massage combined with Chinese medicine group (both P < 0.05). Conclusion The treatment of infantile anorexia with infantile massage and Chinese medicine based on the thought of invigorating spleen and nourishing qi has a significant effect.

BACKGROUND: Studies have confirmed that titanium dioxide coating can improve the adhesion of implant surface, promote osteoblast proliferation and other cell functions, and improve the bone integration ability between the surrounding bone and implant. But little has been done in the use of titanium dioxide coating as a coating for internal and external fixation to prevent screw loosening. OBJECTIVE: To prepare titanium dioxide coated titanium screws and to observe the effect of titanium dioxide coated titanium screws used for bone fixation to reduce screw loosening. METHODS: Titanium dioxide coating was prepared on the surface of titanium screw by anodic oxidation. Thirty male New Zealand white rabbits (provided by the Central Laboratory of Shanghai Putuo District Central Hospital in China) were randomly divided into two groups (n=15 per group). Titanium dioxide coated screws were implanted in the right tibia of the rabbits in the experimental group, and ordinary titanium screws were implanted in the right tibia of the rabbits in the control group. Bone tissue around the screw canal was taken at 1 day, 1 and 4 weeks after implantation, and levels of platelet-derived growth factor and vascular endothelial growth factor genes were detected using RT-PCR. Four weeks after implantation, bone tissue around the nail canal was taken for hematoxylin-eosin staining. RESULTS AND CONCLUSION: (1) The results of hematoxylin-eosin staining showed that inflammatory cell infiltration was seen in the screw canal of the control group, and some screw canal threads were destroyed and absorbed. In the experimental group, the screw threads were clear and had no damage or absorption; inflammatory cells in the screw canal were less exuded. (2) RT-PCR analysis revealed no significant difference in the expression of platelet-derived growth factor and vascular endothelial growth factor gene between the two groups at 1 day after implantation (P> 0.05), while the expression of platelet-derived growth factor and vascular endothelial growth factor gene in the experimental group was significantly higher than that in the control group at 1 and 4 weeks after implantation (P < 0.05 or P < 0.01). Therefore, titanium dioxide coated titanium screws are more biocompatible than common titanium screws and can effectively prevent screw loosening when used for internal fixation.

Objective To study the role of protective antigen(PA)and N-terminal segment of lethal factor (LFn)in the entrance of EGFP(enhanced green fluorescent protein)into HeLa cells. Methods The DNA fragments encoding LFn and EGFP were amplified,respectively,and cloned into the plasmid pET-21 a(+)one after another to construct a recombinant plasmid pET-LFn-EGFP. The plasmid was txansformed into BL21 cells to express LFn-EGFP protein under the induction of IPTG. The protein was purified by Ni chelating chromatography. After incubation with LFn-EGFP in the presence of PA or not, the HeLa cells were analyzed by flow cytometry or laser confocal microscopy. Results The fusion protein LFn-EGFP was purified by over 90% homogeneity and retained the ability of LF to bind with PA when incubated with J774A.1 macrophage cells,and could get into HeLa cells. Conclusion The LFn-EGFP could enter the HeLa cells in a PA independent pathway. But PA could help more LFn-EGFP molecules enter into HeLa cells.

This study was aimed to discuss the influence of specific acupoint acupuncture therapy to the serum angiogenesis factor of high fat diet obese mouse models. Mice were randomly divided into the blank control group , acupuncture control group , model control group , and acupuncture treatment group . There were 6 mice in each group . Obese mouse models were induced after 15-week high fat diet . The specific acupoint acupunc-ture therapy was used as an intervention treatment method for 10 days . The enzyme-linked immunosorbent as-say (ELISA) was used in the detection of serum insulin, vascular endothelial growth factor (VEGF), soluble vascular endothelial growth factor receptor , leptin and the level of nitric oxide in mice . The results showed that the serum insulin , nitric oxide and leptin level in the obese mouse models were increased . However , there were no obvious changes on the vascular endothelial growth factor and soluble vascular endothelial growth fac-tor receptor level . The specific acupoint acupuncture therapy can obviously reduce the level of serum nitric ox-ide and leptin, and improve the content of vascular endothelial growth factor in obese mouse models. However, there were no influence on the level of blood glucose and soluble vascular endothelial growth factor receptor . It was concluded that the specific acupoint acupuncture therapy method had preferable antiobesity action . Its mechanism of action may be related to the regulation of angiogenesis .

Objective To investigate the expression of intestinal epithelial stem cell markers Msil and Hesl in mouse acute radiation enteritis (RE) model. Methods Forty female BALB/c mice were undergone one-time whole abdominal irradiation by 300 cGy/min β-ray. Thirty mice were selected before irradiation (Day 0) and every six mice were euthanized at day 3,5,7 and 14 after irradiation. The small intestinal were detached and the expression of Msil and Hesl were detected in middle part of small intestinal by quantitative RT-PCR, Western blots, and immunohistochemistry. Subsequently,the difference of Msil and Hesl expression was compared. Results The mouse acute RE model was successfully established. At the 5th day after radiation, the results of quantitative RT-PCR indicated that the relative ΔCt value of Msil and Hesl expression were 15. 17 ± 0. 47 and 15. 83 ± 0. 24respectively, the results of Western blot showed that the integrated intensity were 0. 443±0. 055 and 0. 301 + 0. 047 for Msil and Hesl. The expressions were significantly higher than other time points at RNA and protein levels (P<0. 05). The results of immunohistochemistry revealed that Msil and Hesl were highly expressed in intestinal crypts at 5th day and 7th day. Conclusions The expression of Msil and Hesl, upregulated before wound repair in RE model, indicates that intestinal epithelial repair is related to intestinal stem cell proliferation.

OBJECTIVETo investigate the effects of injecting Shenfu, an extract of traditional Chinese herbal medicines, on the central nervous system (CNS) and the cardiac toxicity of bupivacaine in rats.

METHODSSixteen male Sprague-Dawley rats, weighing form 280 to 320 g, were randomly assigned to two groups (n = 8 in each group). Animals in the control group received a saline injection 10 ml/kg while animals in the Shenfu group received an injection of Shenfu 10 ml/kg intraperitoneally 30 minutes before intravenous infusion of bupivacaine. Lead II of an electrocardiogram (EEG) was continuously monitored after 3 needles were inserted into the skin of both forelimbs and the left hind-leg of each rat. The femoral artery was cannulated for measurement of arterial blood pressure and blood sampling. The femoral vein was cannulated for the infusion of bupivacaine. After baseline measurement (arterial blood pressure, heart rate and arterial blood gas), 0.5% bupivacaine was infused intravenously at a rate of 2 mg.kg(-1).min(-1) to all animals until asystole occurred. The time of bupivacaine-induced convulsions, arrhythmia and asystole were determined. The dose of bupivacaine was then calculated at the corresponding time point.

RESULTSThe doses of bupivacaine that induced convulsions, arrhythmia and cardiac arrest were remarkably larger in Shenfu injection-treated animals than in saline-treated rats [convulsions, (10.5 +/- 1.9) mg/kg vs (7.2 +/- 1.5) mg/kg; arrhythmia (10.5 +/- 2.0) mg/kg vs (7.2 +/- 1.9) mg/kg; asystole, (32.8 +/- 8.5) mg/kg vs (25.0 +/- 5.0) mg/kg; P = 0.006, 0.009 and 0.044, respectively].

CONCLUSIONThe Shenfu injection is able to reduce the toxicity of bupivacaine to CNS and cardiac system in rats.

Anesthetics, Local ; toxicity ; Animals ; Bupivacaine ; toxicity ; Drugs, Chinese Herbal ; administration & dosage ; pharmacology ; Electrocardiography ; drug effects ; Heart Rate ; drug effects ; Injections, Intraperitoneal ; Male ; Random Allocation ; Rats ; Rats, Sprague-Dawley

Objective To establish a smooth three-dimensional(3D)geometric model of the maxilla based on CT data using four dif-ferent software packages,to mimic the modified Le Fort Ⅰ osteotomy and its fixation scheme,and to perform a finite element analysis of the postoperative occlusion.Methods CT data were preliminarily processed using Mimics software to produce an STL 3D model.The model was then imported into Inspire Studio software to create a smoothed PolyNURBS geometric model.SpaceClaim software was used to model the surgical osteotomy and fixation schemes.Finally,ANSYS Workbench was used to conduct a 3D finite element analysis simu-lating the patient's occlusion after surgery.Results The simulation results showed that the connection relationship of the finite element model was accurately established under the molar occlusion condition.Under a total occlusal force of 6 N,the maximum equivalent stress of the titanium plate was 73 MPa.Conclusion The maxillary modeling and analysis method used in this study can produce a smooth geometric model suitable for finite element simulation.The results of this study can provide reference for various fixation schemes in orthognathic surgery.