Objective To investigate the mechanism of implanted tissue-engineered bone (TEB)recruiting endogenous mesenchymal stem cells (BMSCs) towards bone regeneration after traumatic bone defect.Methods In vivo experiments:2 mm of diaphysis and periosteum were removed from the middle of the femoral shaft in 8 week old FVB/N mice to form a large segment of bone defect.Demineralized bone matrix (DBM) and TEB were implanted into the defect area and fixated.All mice were randomly divided into DBM group (n =18) and TEB group (n =18).The results were observed 24 hours after implantation:(1) flow cytometry was used to evaluate the number of mobilized host BMSCs into the blood;(2) non-invasive bioluminescent imaging was used to observe the ability of two groups in recruiting mouse bone marrow derived mesenchymal stem cells (mBMSCs) in peripheral blood to the defect area;(3) ELISA was used to evaluate the stromal cell-derived factor 1 (SDF-1) content in peripheral blood of two groups.In vitro experiments:(1) transwell assay was conducted to evaluate the ability of SDF-1 (100 ng/ml) in promoting the migration of human bone marrow derived mesenchymal stem cells (hBMSCs).SDF-1/C-X-C motif chemokine receptor-4 (CXCR4) pathway was blocked by the selective CXCR4 antagonist Plerixafor (AMD3100).The experimental groups were divided into control group,SDF-1 group,and SDF-1 + AMD3100 group.(2) The co-culture system of human umbilical vein endothelial cells (hUVECs) and hBMSCs was established,and cells were stimulated by SDF-1.The experimental groups were divided into hBMSCs group,hBMSCs + hUVECs group,and hBMSCs + hUVECs (AMD3100 pretreatment) group.Transwell assays were used to compare the migration of hBMSCs in each group.ELISA was used to detect the concentration of hepatocyte growth factor (HGF) in the co-culture supernatant.(3) In vitro cultured hUVECs were stimulated by SDF-1 and SDF-1/CXCR4 pathway was antagonized by AMD3100.The experimental groups were divided into control group,SDF-1 group,and SDF-1 + AMD3100 group.Quantitative real-time polymerase chain reaction (qRT PCR) was used to evaluate the expression of HGF in each group.Results In vivo experiments:24 h after transplantation,the number of BMSCs and SDF-1 concentration in the TEB group were significantly highcr than those in the DBM group (P ＜ 0.05).The number of recruited mBMSCs into the circulation in the TEB group was larger than that in the DBM group (P＜ 0.01).In vitro experiments:(1) compared with the control group and the SDF-1 + AMD3100 group,the SDF-1 group significantly enhanced the migration ability of hBMSCs in Transwell migration experiments (P ＜ 0.01);(2) compared with the hBMSCs group and the hBMSCs + hUVECs (AMD3100 pretreatment) group,the number of migrated cells and HGF concentration in the hBMSCs + hUVEC group significantly increased (P ＜ 0.01),but there were no significant differences between the hBMSCs group and the hBMSCs + hUVECs (AMD3100 Pretreatment) group (P ＞0.05);(3) qRT-PCR showed that the expression of HGF was significantly increased in the SDF-1 group compared with the control group (P ＜ 0.05).After antagonizing SDF-1/CXCR4,HGF expression in the SDF-1 + AMD3100 group was significantly lower than that in the SDF-1 group.Conclusions TEB transplantation in traumatic bone defect can significantly increase the concentration of chemokine SDF-1 in vivo and effectively promote the mobilization of endogenous MSCs and recruitment of circulating MSCs.SDF-1 not only directly promotes the migration of hBMSCs through SDF-1/CXCR4 pathway,but also up-regulates the expression and secretion of HGF in vascular cells to further amplify the chemotactic effect of SDF-1 on hBMSCs.

Objective Toidentify factors contributing to posthepatectomy liver dysfunction (PHLD),focusing on the Functional remnant liver volume to spleen volume ratio (FreLSVR).Methods The clinical data of 74 patients undergoing precise liver resection from January 2016 to October 2017 were retrospectively analyzed.IQQA liver system was used to reconstruct the liver and spleen 3D image by using patients' preoperative abdominal CT image data.Tumor volume,3D estimated functional residual liver volume,spleen volume and FreLSVR were measured and calculated.Preoperative and postoperative liver function test,blood coagulation function test,operation time,intraoperative blood loss,and the volume of daily postoperative abdominal drainage were recorded.Correlations between multiple parameters and PHLD were analyzed.Results PHLD occurred in 16 (21.6％).Single factor analysis revealed that the standardized residual liver volume ratio (P＜0.05),FreLSVR (P＜0.05) and preoperative AST value (P＜0.05) were correlated with postoperative hepatic insufficiency.Multivariate regression analysis showed that FreLSVR (OR=0.535,95％CI=0.305～0.936,P＜0.05) was the only independent factor of PHLD.In the ROC curve analysis for FreLSVR,a cut-off value of 2.56 (AUC=0.824,Sensitivity 81.1％,specificity 71.7％.) was the appropriate value for predicting the risk of PHLD according to Youden index.Then the patients were regrouped according to this cut-off value.Compared with the FreLSVR＞2.56 group (53 cases),the highest postoperative INR value (1.46± 0.19 to 1.29± 0.29,t=-2.405,P＜0.05) was higher that of FreLSVR≤2.56 group,and the amount of average daily abdominal drainage in one and two weeks after operation was higher,(188.0(79.2 ～ 375.1)ml to 96.0(46.5 ～ 179.3)ml,P＜0.05) and (207.2(125.6 ～ 827.1)ml to71.8(14.0 ～ 179.8) ml,P＜0.05),respectively.Conclusion FreLSVR has significant correlation with postoperative hepatic dysfunction,and provides guidance for the safety of liver resection in the future.

s:Apoptosis, also known as programmed cell death, is a gene controlled active cell suicide process in order to maintain balance of organisms. Apoptosis is mainly mediated through three signal pathways, including the endoplasmic reticulum, mitochondria, and death receptor. As a kind of naphthoquinone compounds and one of the main active ingredients of Lithospermum, shikonin has many biological activities, and its anti-cancer research is recent hot spot. The anti-cancer mechanism of shikonin is closely related with apoptosis and three kinds of signaling pathways. This article reviewed the research progress in apoptosis and signal pathways induced by shikonin.

OBJECTIVE:To optimize the water extraction technology of Yangxue Jiegu capsule. METHODS:Using adding amount of water,decoction time and decoction times as investigation factors,ferulic acid content,icariin content and yield rate as investigation indexes,orthogonal test was used to optimize the optimal water extraction technology of Yangxue Jiegu capsule;and verification test was conducted. RESULTS:The optimized extraction technology was 10-fold water,extracting 3 times,1 h once. Contents of ferulic acid and icariin were 0.38,1.23 mg/g(n=3),yield rate was 28.2%(n=3),average comprehensive score was 97.11 (RSD=2.77%,n=3). CONCLUSIONS:Optimized extraction technology is feasible,accurate,reliable,and can be used for the extraction and preparation of Yangxue Jiegu capsule.

Objective To development a cooling crystallization process that is suitable for industrial preparation of purified dihydromyricetin.Methods Screen design was used to investigate effects of process parameters such as,temperature,concentration ethanol aqueous,quantity of activated charcoal and adsorption time on yield and purity of dihydromyricetin.Purity was verified by high performance liquid chromatography and thin layer chromatography.The solubility of dihydromyricetin in water at viable temperature and ethanol proportion was also determined by UV spectrophotometry.The solid form was characterized by thermal analysis and powder X-ray diffractometry.Results When temperature was ＞ 85 ℃,ethanol concentration ＜ 10％,dosage of activated charcoal 0.1％-0.3％,and adsorption time 1-3 min,yield of dihydromyricetin was more than 70％,and the purity greater than 98％.The crystals,prepared by cooling crystallization from water and ethanol aqueous,had the same physical form and crystal habit.Conclusion Cooling crystallization from low concentration of ethanol aqueous gets higher yield and the process is more robust than crystallization from water.

Objective Cerebral microbleeds (CMBs) are important indicators of cerebral small vessel disease .However, it is still unclear whether endothelial dysfunction is involved in CMBs .The aim of this study is to investigate the correlation between CMBs and soluble E-selectin (sE-selectin) in patients with acute ischemic stroke . Methods Based on the results of MRI (3.0 T) susceptibility weighted imaging , we divided patients with first acute ischemic stroke into a CMBs group ( n=63 ) and a non-CMBs group (n=63), and recruited another 45 volunteers with normal MRI findings as controls .We collected and conducted comparative a-nalysis on the demographic data , biochemical variables ( including the sE-selectin level ) , vascular risk factors , and the number of CMBs of the patients . Results Ordinal logistic regression analysis showed a significantly positive correlation between sE -selectin and the number of CMBs (OR=1.062, 95%CI:1.023-1.103, P=0.002), higher systolic blood pressure associated with more CMBs (OR=1.014, 95%CI:1.002-1.025, P=0.021). Conclusion Serum sE-selectin is significantly positively correlated with and can be used as a biological marker for the severity of CMBs .

Objective To explore the effects of volatile oil and 2-undecanone from Houttuynia Cordata Thunb. (H. cordata) on LPS-TLR4 / MD-2-TNF-α signaling pathway. Methods TLR4 / MD-2 blocking agent was used to mask the TLR4 /MD-2 site,then protein expression levels of TLR4 in cells treated with volatile oil and 2-undecanone were analyzed by western blot. ELISA was used to detect the secretion of the inflammatory cytokines such as TNF-α,IL-1β and IL-10. Comparison analysis was then performed from the results of cell experiments in vitro and anti-inflammatory effects through xylene-induced ear edema test in vivo. Results In concentrations between 1 to 10 μg · mL-1 ,Houttuynia volatile oil showed better effect than 2-undecanone on inhibition of TLR4 protein in LPS-induced RAW264. 7 cells, and had some differences in the effects on inflammatory factors. Compared with the LPS+TLR4 / MD-2 group,the LPS+TLR4 / MD-2 + volatile oil group had no significant difference in the expression of TLR4 protein (P>0. 05),but the LPS+TLR4 / MD-2 +2-undecanone group reduced the expression of TLR4 protein obviously. It appeared that volatile oil exerts its anti-inflammatory effect through LPS-TLR4 / MD-2-TNF-αpathway,but 2-undecanone may exert its anti-inflammatory effect by other means. Houttuynia volatile oil showed better anti-inflammatory activity than 2-undecanone in vivo at the same dose. Conclusion There are some differences in anti-inflammatory effects and related mechanisms between volatile oil and 2-undecanone, probably owing to the synergistic effects of multi-ingredients in the volatile oil.

Objective To analyze the variations of glycerol-3-phosphate dehydrogenase 1 like gene (GPD1-L) and address the association with sudden m anhood death syndrom e (SMDS). Methods The genom ic DNA was extracted from blood sam ples of the SMDS group and the norm alcontrolgroup.The exons, exon-in-tron boundaries and 3′-U TRs of coding region of GPD1-L w ere PCRam plified and DNAsequenced di-rectly to confirm the types of variations. The genotype frequency and allele frequency w ere analyzed statistically. Results There w ere tw ovariants in the SMDS group, c.465C>Tand c.*18G>T, the latter existed certain degree difference of genotype distribution and allele frequency betw een the SMDS group and the control group, but there was no statistically significant (P>0.05). Conclusion The relation be-tw een gene m utation of GPD1-L and the occurrence of Chinese SMDS deserves a further research.

Currently endoscopic retrograde cholangiopancreatograhp(ERCP) has been one of the major treatments to diagnose and cure pancreatic and choledochal diseases in many large and medium-sized hospitals.To meet the requirements of the army,ERCP treatment is practiced in field battle X-ray diagnosis vehicle(Model XCY2002-1/200) and has achieved satisfactory effects.The functions of field battle X-ray diagnosis vehicle in model of XCY2002-1/200 are expanded and new access are explored to cure patients with pancreatic and choledochal diseases in the field.