Objective To study the effect of hyperthermia-induced MMP-9 on expression of claudin-1 in in vitro BBB model.Methods BMEC and astrocytes were cultured.An in vitro BBB model was established with its permeability assayed by LY test.The BBB model was randomly divided into 37℃ group,39℃ group,37℃+MMP-9 group and 39℃+MMP-9 group.The permeability of BBB model was recorded at 30,60 and 90 min respectively.Expression of claudin-1 in BMEC was detected by Western blot with immunocytochemical staining.Results The permeability of BBB model was higher in 39℃ group than in 37℃ group (P＜0.05) and was significantly higher in 39℃ +MMP-9 group than in 37℃ group (P＜0.05).The number of claudin-1 positive BMEC was smaller in 39℃ +MMP-9 group than in 37℃ group at the same time points (P＜0.05) and in 39℃ +MMP-9 group than in 39℃ group at 90 min (75.0±12.8/HP vs 97.0±6.1/HP,P＜0.05).The expression level of claudin-1 was lower in 39℃ group than in 37℃ group (P＜0.05) and was significantly higher in 39℃ +MMP-9 group than in 37℃ group (P＜0.05),which further increased with the increasing time.Conclusion Hyperthermia can downregulate the expression of claudin-1 in in vitro BBB model and upregulate the permeability of in vitro BBB model.Adding MMP-9 can further aggrevate the damage of in vitro BBB model,indicating that hyperthermia can aggrevate the damage of in vitro BBB model through MMP-9.

[Objective]To observe the growth status and osteogenic potential of gene transferred mesenchymal stem cells with Ad-BMP-2 planted on allogenetic acellular and decalcified bone matrix served as tissue engineering scaffold.[Method]Rabbit mesanchymal stem cells transferred with Ad-BMP-2 were seeded on allogenetic acellular and decalcified bone matrix,the growth status of gene transferred MSCs on the scaffold was observed by scanning electron microscopy and the ostengenic potential was observed by testing contents of ALP and BGP and secretion of Collagen I in the culture supermatant.[Result]The MSCs transfected with Ad-BMP-2 grew well on the tissue engineering scaffold,the osteogenic ability of the gene transfected MSCs showed great significant difference compared with that of the control group.[Conclusion]The MSCs transfected with Ad-BMP-2 can grow well on the acellular and decalcified bone matrix and the osteogenic potential can be great improved by gene transfection.

County hospitals occupy an important place in the delivery of community health service as they are the center of technological guidance, the base for training in general medicine, and a coordinator of and participant in the community health service of the county seats. To do a good job of community health service, county hospitals ought to make efforts in the following aspects: restructuring of human resources, cooperation between various community health institutions, diversified training programs for personnel engaged in community health service and doctors at the grass-roots level, transformation of ideological concepts, and updating of knowledge and skills.

The pathophysiological mechanism of ischemic stroke is very complex.Many intracellular and extracellular factors play the important roles in it.Studies have shown that the expression of matrix metalloproteinase-9 (MMP-9) has the changes of quantity,time and space after stroke.These changes are closely associated with the occurrence and development of ischemic stroke.The experimental studies focusing on MMP-9 also have made corresponding effects.The in-depth study of the mechanism of action of MMP-9 is expected to provide a new approach for the diagnosis and treatment of ischemic stroke.

AIM:To establish a method for detecting three aconitum alkaloids(aconitine,mesaconitine,hy-paconitine) in Zhentong Huoluo Tincture(Radix Aconiti,Radix Aconiti kusnezoffii,Radix et Rhizoma Rhei,Rhizoma Pinelliae,Rhizoma Arisaematis ect.).METHODS:A ZOBAX Extend-C_ 18(250 mm?4.6 mm,5 ?m) column was used to determine the aconitum alkaloids in Zhentong Huoluo Tincture.The mobile was gradient elution,0.1% die-thylamine(adjust pH 9.0 with phosphoric acid) as mobile A;0-20 min A∶B(63∶37),20-45 min A∶B(63∶55∶37→45),at the rate of 1 mL/min.The temperature of column was 30 ℃,and the wavelength was at 232 nm.RESULTS:The linearity of the aconitine in 9.74 ng-974 ng was 0.999 9;that of the mesaconitine in 23.86-2 386 ng was 0.999 9 and that of the hypaconitine in 29.94 ng-2 994 ng was 0.999 9.The RSD of precision for these three were 1.50%、0.68% and 0.49%,respectively.The average recoveries and its RSD were 100.2%(RSD 3.2 %)、102.9%(RSD 2.2%) and 98.2(RSD 2.0%),respectively for aconitin,mesaconitine and hypaconitine.The reproducibilities were all less than 2.0%,and the sample solution was stable in 24 h.CONCLUSION:The method is simple,accurate and has good stability,can be used for the quantity control in the product of Zhentong Huoluo Tincture.

AIM: To establish a method for detecting the residual solvents in protopanaxdiol. METHODS: Capillary GC with HP-PLOT-Q column (30 m ? 0. 53 mm ? 1. 0 ?m) was used to determine residual in protopanaxdiol. N,N-dimethyl-acetamide as solvent media,FID detector and nitrogen as the carrier gas. RESULTS: There was a good linearity at the experimental concentration (r = 0. 999 96-0. 999 99),the RSD of precision was less than 6. 0 % . The spotting recovery of the 3 residul,including methol,ethyl acetate and n-butanol,was in the range of 98. 2%-102. 2% ,and its’RSD was 1. 1%-2. 4% . CONCLUSION: The method is simple,accurate and of high sensitivity,can be used for the determination of residual solvents in protopanaxdiol.

Objective To evaluate the effect of dexmedetomidine on the inflammatory responses in brain tissues in septic rats.Methods Seventy-two male Sprague-Dawley rats,aged 10 weeks,weighing 250-280 g,were randomly divided into 4 groups (n =18 each):control group (group C); sepsis group (group lipopolysaccharide (LPS)) ; distilled water group (group DW) and dexmedetomidine group (group D).Sepsis was induced by intraperitoneal injection of LPS 5 mg/kg (dissolved in normal saline 1 ml) in groups LPS,DW and DEX,while normal saline 1 ml was injected intraperitoneally in group C.Distilled water 20 μl was injected into the lateral cerebral ventricle in group DW,while dexmedetomidine 3 μg/kg (dissolved in distilled water 20μl) was injected into the lateral cerebral ventricle in group DEX.Six animals were sacrificed at 1,2 and 6 h after administration and hippocampi were removed for determination of TNF-α and IL-6 contents (by ELISA) and TLR4 mRNA expression in hippocampal tissues (by RT-PCR).Results Compared with group C,TNF-α and IL-6 contents in hippocampus tissues were significantly increased at each time point after administration in group LPS (P ＜ 0.05).Compared with group LPS,no significant change was found in TNF-α and IL-6 contents in hippocampal tissues (P ＞ 0.05),and TLR4 mRNA expression was significantly up-regulated at each time point after administration in group DW (P ＜ 0.05).Compared with group DW,TNF-α and IL-6 contents in hippocampal tissues were significantly decreased at each time point after administration,and TLR4 mRNA expression was significantly up-regulated at 2 and 6 h after administration in group DEX (P ＜ 0.05).Conclusion Dexmedetomidine can reduce inflammatory responses in brain tissues in septic rats via down-regulating TLR-4 mRNA expression.

Objective To explore the effects of electrical stimulation in different acupoints on the expression of growth associated protein-43 (GAP-43), basic fibroblast growth factor (bFGF), insulin-like growth factor (IGF) in spinal cord injured rats after the human umbilical cord blood stem cells transplantation.Methods 192 SD rats were injured at T10-11 level with NYU, and divided randomly into groups: Group A received electric stimulation in the scalp of motor area (A1 group received electric stimulation and transplantation of umbilical cord blood stem cell, the rats of A2 group only received electric stimulation); group B received local electric stimulation at damaged site (B1 with electric stimulation and transplantation, B2 only with electric stimulation); group C received electric stimulation in the scalp of motor area and at damaged site (C1 with electric stimulation and transplantation, C2 only with electric stimulation); D1 received transplantation without electric stimulation, D2 neither with transplantation nor electric stimulation. The expression of GAP-43, bFGF and IGF were detected 1, 2, 3, 4, 8 and 12 weeks after operation. Results Electric stimulation increased the expression of GAP-43, bFGF, IGF, stimulating scalp and body were more than stimulated scalp or body alone, combined with the stem cells transplantation were more than no transplantation. Conclusion Both electric stimulation and stem cells transplantation can improve the microenvironment for neural recovery synergistically. Stimulating scalp and body is more effective than alone.

AIM:To investigate the role of peripheral blood leukocytes and polymorphonuclear neutrophils (PMNs) on intestinal injury following mesenteric ischemia/reperfusion (IR) in rats. METHODS: Twenty adult, male Sprague-Dawley rats, weighing 200-230 g, were randomly divided into two groups. The control group (CON) consisting of 10 rats was subjected to laparotomy and separation of superior mesenteric artery (SMA) only. The ischemia/reperfusion (IR) group consisting of 10 rats, was subjected to laparotomy, followed by occlusion of the superior mesenteric artery (SMA) by an atraumatic microvascular clamp for 30 min. At the end of ischemic period in IR, the microvascular clamp was removed and the intestinal segment was reperfused for 60 min. The pathological changes of the ileal mucosal tissue were evaluated. The apoptosis of intestinal mucosal epithelial cells was examined by terminal deoxylnucleotidy-l transferase mediated-dUTP nick end-labeling (TUNEL). The enzymatic activity of casapse-3 in mucosal cells was determined using a colorimetric assay. The percentages of apoptotic peripheral blood leukocytes and PMNs were measured by flow cytometry using Annexin-V/PI double staining assay. The numbers of peripheral blood leukocytes in each animal was measured at baseline, 30 min of ischemia, and 30 min and 60 min of reperfusion. RESULTS: (1) Compared to CON group animals, the most severe mucosal injury was observed in IR group under optical microscope. (2) The number of apoptotic mucosal epithelia cells and enzymatic activity of caspase-3 were significantly higher in IR than those in CON group (P