[Objective]To study the combination of calcium phosphate cement(CPC) and morselized bone and their optimum mixing ratio. [Methods]Bilateral nemora of macro-ears rabbits were made into morselized bone.There were two groups:experimental group and control group.In the experimental group,the ratios of morselized bone to CPC were 1 to 1,1 to 2,1 to 3,1 to 4 and 1 to 5.In the control group CPC was used after it was solidified.The maximum compression strength and solidified time of the materials in experimental and control groups were tested.The cell multiplication was evaluated by MTT after mesenchymal stem cells were cultivated.[Results]The cell multiplication in materials mixed with ratios of 1 to 2 and 1 to 3 was better than those in the materials mixed with the other ratios by MTT.It indicated that there was a significant difference(P

[Objective]To confirm the effect of combination of calcium phosphate cement and small amounts of autoallergic micromorselized bone for reparing bone defect and provide the basis for clinical use.[Method]The animal models of bilateral radius defect were created by surgery on Japan white big ear rabbits.The combination with a ratio of 3 to 1 in one lateral leg was implanted in experimental group,and calcium phosphate cement was used in the control group.The X-rays,bone density,and maximum torsional moment were carried out at different time points,and osteogenesis and degradation of the materials in the two groups were analyzed.[Result]Enhancement of osteogenesis was found in the experimental group,and the materials were enclosed significantly by bony callus at the 8~(th) week,the defects were repaired and the materials were degraded completely at the 16~(th) week.The bone mineral content in the experimental group was higher than that in the control group(P

Vascular smooth muscle cell (VSMC) proliferation and migation play important roles in the occurrence and development of atherosclerosis and restenosis after intravascular stenting.The studies in recent years have shown that kallikrein-kinin system (KKS) is closely correlated with VSMC proliferation and migration in cytokines and transduction pathways.Therefore,investigating the roles of KKS in the VSMC proliferation and migation process has great significance in clinical prevention and treatment of atherosclerosis and restenosis after intravascular stenting.

Objective To investigate the effects of Sulfotanshinone Sodium Injection (SSI) on neuropathic pain in rats.Methods One hundred and eight adult male Wistar rats,aged 6-8 weeks,weighing 180-220 g,were randomly divided into 3 groups (n =36 each):sham operation group (group S) ; chronic constrictive injury (CCI)group; group SSI.The animals were anesthetized with intraperitoneal 10% chloral hydrate 350 mg/kg.In groups CCI and SSI,4 ligatures were placed on the right sciatic nerve at 1 mm intervals with 4-0 silk thread according to the method described by Bennett et al.In group S,the right sciatic nerves were exposed,but not ligated.In group SSI,SSI 25 mg/kg was injected intraperitoneally once a day starting from the end of operation until one day before the animals were sacrificed,while the rats received the equal volume of normal saline (5 ml/kg) instead of SSI in groups S and CCI.Twelve animals in each group were chosen at 1 day before operation and 3,7 and 14 days after CCI (T1-4) to measure mechanical paw withdrawal threshold to yon Frey stimuli (MWT) and paw withdrawal latency to thermal nociceptive stimulus (TWL).Six rats in each group were sacrificed at T2-4 after measurement of pain threshold,and their lumbar segnents (L4-6) of the spinal cord were immediately removed for determination of Bcl2 and caspase-3 expression in spinal dorsal horn (by immune-histochemistry),and MDA content and SOD activity (by spectrophotometry) in spinal cord.Results Compared with group S,PWT was significantly decreased,PWL was shortened,the expression of Bcl-2 and caspase-3 was up-regulated,MDA content was increased and SOD activity was decreased at T2-4 in groups CCI and SSI (P ＜ 0.05).Compared with group CNP,PWT was significantly increased,PWL was prolonged,the expression of Bcl-2 was up-regulated,the expression of caspase-3 was downregulated,MDA content was decreased and SOD activity was increased at T2-4 in group SSI (P ＜ 0.05).Conclusion SSI can mitigate neuropathic pain in rats and inhibition of oxidative stress in spinal cord tissues and reduction of apoptosis in spinal dorsal horn neurons are involved in the mechanism.

Objective To evaluate the effect of dexmedetornidine on the expression of Toll-like receptor 4 (TLR4) and nuclear factor kappa B (NF-κB) in the spinal cord in rats with neuropathic pain (NP).Methods One hundred and eight male Wistar rats,aged 6-8 weeks,weighing 180-220 g,were randomly assigned into 3 groups (n =36 each):sham operation group (group S),NP group and dexmedetomidine group (group D).NP was induced by chronic constrictive injury in anesthetized rats.Sciatic nerve was exposed and 4 loose ligatures were placed on the sciatic nerve at 1 mm intervals with 4-0 silk thread.In group S,the right sciatic nerves were exposed,but not ligated.Dexmedetomidine 50 μg/kg was injected intraperitoneally once a day from the onset of operation to one day before the rats were sacrificed in group D,while the equal volume of normal saline was injected in groups S and NP.Mechanical withdrawal threshold (MWT) and thermal pain threshold (TPT) were measured on the day before operation (T0) and 3,7,and 14 days after operation (T1-3).After measurement of pain threshold at T1,T2 and T3 after operation,the L4-6 segments of the spinal cord were removed for determination of the expres-sion of TLR4 and NF-κB mRNA (by RT-PCR) and the expression of TLR4 and NF-κB in spinal dorsal horn (by immuno-histochemistry).Results Compared with group S,MWT and TPT were significantly decreased and the expression of TLR4,NF-κB and TLR4 and NF-κB mRNA was up-regulated after operation in groups NP and D (P ＜ 0.05).Compared with group NP,TPT and MWT were significantly increased and the expression of TLR4,NF-κB,TLR4 mRNA and NF-κB mRNA was significantly down-regulated after operation in group D (P ＜ 0.05).Conclusion The mechanism by which dexmedetomidine attenuates NP in rats is related to inhibition of the expression of TLR4 and NF-κB in rat spinal cord.

Objective To evaluate the effects of dexmedetomidine on the activity of cAMP response element binding protein (CREB) and c-fos in the spinal dorsal horn in a rat model of neuropathic pain.Methods Fifty-four adult male Wistar rats,aged 6-8 weeks,weighing 180-220 g,were randomly divided into 3 groups (n =18 each):sham operation group (group S),chronic neuropathic pain group (group C) and dexmedetomidine group (group D).The animals were anesthetized with intraperitoneal 10％ chloral hydrate 350 mg/kg.The sciatic nerve was exposed and 4 ligatures were placed on the right sciatic nerve at 1 mm intervals with 4-0 silk thread in C and D groups.In group D,dexmedetomidine 50 μg/kg was injected intraperitoneally once a day starting from the end of operation until 1 day before the animals were sacrificed,while the equal volme of normal saline was injected instead of dexmedetomidine in S and C groups.Paw withdrawal threshold to mechanical stimulation with yon Frey filament (MWT) and paw withdrawal latency to thermal stimulation (TWL) were measured on 1 day before operation and 3,7 and 14 days after operation.The animals were sacrificed after measurement of MWT and TWL.Their lumbar segments (L4-6) of the spinal cord were removed for measurement of the expression of phosphorylated CREB (pCREB) and c-fos by immunohistochemistry.Results Compared with group S,MWT was significantly decreased,TWL was shortened,and the expression of pCREB and c-fos was up-regulated on 3,7 and 14 days after operation in C and D groups (P ＜ 0.05).Compared with group C,MWT was significantly increased,TWL was prolonged,and the expression of pCREB and c-fos was down-regulated on 3,7 and 14 days after operation in group D (P ＜ 0.05).MWT was significantly lower,and TWL was shorter on 3,7 and 14 days after operation than on 1 day before operation in C and D groups (P ＜ 0.05).MWT was significantly lower,TWL was shorter,and the expression of pCREB and c-fos was higher on 7 and 14 days after operation than on 3 days after operation in C and D groups (P ＜ 0.05).MWT was significantly higher,TWL was longer,and the expression of pCREB and c-fos was lower on 14 days after operation than on 7 days after operation in C and D groups (P ＜ 0.05).Conclusion The mechanism by which dexmedetomidine reduces neuropathic pain is related to inhibition of the activity of CREB and c-fos in the spinal dorsal horn of rats.

OBJECTIVE:To establish a method for the qualitative identification and content determination of berberine hydro-chloride in Huangjin paste. METHODS:TLC was adopted for the qualitative identification of berberine hydrochloride and HPLC was conducted for the content determination of berberine hydrochloride in preparation. The column was Symmetry Shield Rp-18 with mobile phase of acetonitrile-0.1% phosphoric acid(50∶50,V/V)at a flow rate of 1.0 ml/min,detection wavelength was 265 nm,column temperature was 30 ℃ and injection volume was 10 μl. RESULTS:TLC spots of berberine hydrochloride in prepara-tion were clear and well-separated. The linear range of berberine hydrochloride was 2.5-20.0 μg/ml(r=0.999 0);RSDs of preci-sion,stability and reproducibility tests were lower than 3%;recovery was 97.7%-102.1%(RSD=1.68%,n=6). CONCLUSI-ONS:The method is simple,accurate and reproducible,and can be used for the qualitative identification and content determination of berberine hydrochloride in Huangjin paste.

Objective To investigate the correlation between ABCD2 scores and vertebrobasilar artery stenosis in patients with posterior circulation transient ischemic attack (TIA).Methods The patients with posterior circulation TIA who performed cerebral angiography were enrolled in the study,and their ABCD2 scores and imaging data were analyzed.Results A total of 108 patients with posterior circulation TIA were enrolled.There were significant differences in the main trunk stenosis degree of vertebrobasilar artery with different ABCD2 scores in patients with posterior circulation TIA (P =0.005).The overall degree of stenosis increased with the increase of ABCD2 score (P =0.000).After adjustment for other vascular risk factors,ABCD2 score had significant predictive ability for whether had posterior circulation stenosis (odds ratio [OR]1.771,95％ confidence interval [CI] 1.234-2.542; P =0.002) and moderate to severe stenosis (OR 2.083,95％ CI 1.336-3.176; P =0.001).Its receiver operating characteristic area under the curve was 0.693(95％ CI 0.586-0.800) and 0.755 (95％ CI 0.661-0.849) respectively.When the ABCD2 score was ≥3,the sensitivity and specificity of predicting posterior circulation stenosis were 62.6％ (95％ CI 53.4％-71.7％) and 70.6％ (95％ CI62.0％-79.1％) respectively,and the sensitivity and specificity of predicting posterior circulation stenosis ＞ 50％ were 78.7％ (95％ CI 70.9％-86.4％) and 67.5％ (95％ CI58.7％-76.3％) respectively.Conclusions In patients with posterior circulation TIA,the stenosis degree of vertebrobasilar arterial trunk increases with the increase of ABCD2 score.In a certain extent,ABCD2 score may predict whether the main trunk of the posterior circulation has stenosis and whether has moderate to severe stenosis.

Objective To evaluate the effect of dexmedetomidine and small dose of ketamine on the expression of P2X4 receptor (P2X4 R) mRNA and P2X7 receptor (P2X7R) mRNA in the dorsal root ganglion of rats with neuropathic pain.Methods Ninety male Sprague-Dawley rats,aged 6-9 weeks,weighing 180-220 g,were randomly divided into 5 groups (n =18 each):sham group (group S),chronic constrictive injury group (group CCI),dexmedetomidine group (group D),ketamine group (group K) and dexmedetomidine + ketamine group (group DK).The animals were anesthetized with intraperitoneal 10％ chloral hydrate 400 mg/kg.Neuropathic pain was induced by CCI in CCI,D,K and DK groups.The sciatic nerve was exposed and 4 loose ligatures were placed on the sciatic nerve at 1mmintervals with 4-0 silk thread.In group S,the sciatic nerves were only exposed but not ligated.In D,K and DK groups,dexmedetomidine 50μg/kg,ketamine 10 mg/kg and dexmedetomidine 25μg/kg + ketamine 5 mg/kg were injected intraperitoneally,respectively,while the equal volume of normal saline was injected in S and CCI groups,once a day for 14 consecutive days after CCI.Mechanical paw withdrawal threshold (MWT) and thermal paw withdrawal latency (TWL) were measured at 1 day before CCI,and 3,7 and 14 days after CCI.Six animals were sacrificed after measurement of pain threshold at 3,7 and 14 days after CCI and the lumbar segments (L4-6) of the dorsal root ganglion were removed for determination of P2X4 R mRNA and P2X7 R mRNA expression by RT-PCR.Results Compared with group S,MWT and TWL were significantly decreased at 3,7 and 14 days after CCI in groups CCI,D,K and DK,the expression of P2X4R mRNA and P2X7R mRNA was up-regulated at 3,7 and 14 days after CCI in groups CCI,D and K,and the expression of P2X4 R mRNA and P2X7 R mRNA was up-regulated at 3 and 7 days after CCI in group DK (P ＜ 0.05).Compared with group CCI,TWL and MWT were significantly increased and the expression of P2X4 R mRNA and P2X7 R mRNA was down-regulated at 3,7 and 14 days after CCI in groups D,K and DK (P ＜ 0.05).Compared with D and K groups,TWL and MWT were significantly increased and the expression of P2X4 R mRNA and P2X7 R mRNA was down-regulated at 3,7 and 14 days after CCI in group DK (P ＜ 0.05).Conclusion The mechanism by which the combination of dexmedetomidine and small dose of ketamine produces a synergistic antinociception in rats with neuropathic pain may be related to down-regulation of the expression of P2X4 R mRNA and P2X7 R mRNA.

Response surface analysis (uniform precision of central composite design, SAS 9.1.3 software) was applied to optimize the four major factors (ratio of soybean meal to water, enzyme quantity, fermentation time and inoculation quantity) for soybean meal solid fermentation. According to the change of the hydrolyzation degree of soybean protein, the equation of polynomial regression was established between those factors and the response. The result showed that the optimum condition included as follows: ratio of soybean meal to water 1∶1.00,enzyme quantity 2.55%, fermentation time 65h and inoculation quantity 1.00%. Under the optimum level, the degree of hydrolyzation reached 13.3%, which increased 56% over pre-optimization.