Objective To demonstrate the histology in acellular dermal matrix(ADM)after being transplanted in vivo over time. Methods Forty male SD rats were recruited for the experiment. Subcutaneous implantation of an 1 cm × 1 cm ADM was given in the left sides on the back of the rat for the experimental group, while only dissection and suturing were performed in right side of the back for the control group. All the animals will be sacrificed at appointed time after operation, Five ADM samples were harvested in each time point. The content and proportion of collagen type were examined with HE staining, Picrosirius staining, Masson′s trichrome staining, and Immunohistochemical staining (targets: pan macrophage, M1 macrophage and M2 macrophage). Results All rats survived after operative without any complications. Significant differences of thickness were not observed at the end of 5 months; HE scores suggested that ADM increased in cell infiltration scores in 2 weeks before the plateau , vascularity also showed a similar trend; Collagen trichrome staining showed a substantial increase in density of collagen bundles with time. The comparison of the proportion of collagen among days showed significant differences (P < 0.05). Immunohistochemical staining of M1 and M2 showed that macrophages had distinct polarization profiles in materials. Furthermore, the comparison of M1 vs M2 response associated with different materials showed significant differences in all time points (P < 0.05). Conclusions The chemically cross-linked ADM could keep long time in the body; ADM significantly stimulated proinflammatory of M2 differentiation from M1 in constructive remodeling.

AIM: To evaluate the alterations of maternal hepatic drug-metabolizing enzymes and antioxidative function in tobacco-induced intrauterine growth retardation (IUGR). METHODS:Pregnant rats were assigned to control group and tobacco group. IUGR model was produced by smoking from gestational days (GD) 9 to GD 20. Fetuses were removed by laparotomy on GD 21. The fetal development parameters such as fetal body weights, litter size and placental weights were recorded. Subcelluar fractions of liver were prepared by differential centrifugation. Activities of drug-metabolizing and antioxidative enzymes were monitored. RESULTS:In the tobacco exposure group, fetal body weights, litter size and placental weights were significantly reduced (P

Objective To investigate the effect of fullerol on the osteogenic differentiation of rat adipose-derived mesenchymal stem cells (rADSCs) based on hanging drop culture. Methods rADSC spheres were formed by hanging drop culture for 3 days, then the spheres were dissociated to single cell by trypsin (called rADSC sphere-derived cells). The rADSC spherederived cells were cultured in 2-D adherent cell cultures for 24 hours, then the ordinary culture medium was replaced by osteogenic induction medium (osteogenic medium, OM). OM without fullerol served as control group (CM); OM with 1.0μmol/L fullerol was used as the experimental group (OM+Ful1.0μmol/L). Two groups were induced to differentiation for 14d and 21d, using two methods of alizarin red staining and quantitative real-time PCR (qPCR) to detecte the ability of rADSC sphere-derived cells differentiating into osteoblasts. Results rADSCs could form a uniform size microspheres structure through hanging drop culture for 3 days; rADSC sphere-derived cells were obtained by dissipation of trypsin. Compared with OM group, OM+Ful1.0μmol/L can make rADSC sphere-derived cells form more mineralized calcium nodules, and enhance the expression of the relevant osteogenic gene Runx2, OCN and ColI. Conclusion Fullerol can significantly enhance the osteogenic differentiation of rADSCs based on hanging drop culture, and it is helpful to improve the efficiency of osteogenic induction of rADSCs.

Objective Using Intelligence Scale of Mini Mental State Estimated (MMSE) as the gold standard to determine the relevance of International HIV-associated Dementia Scale (IHDS)in minority ethnic areas in Guangxi populations with different cultural values.Corresponding boundary value related to the authenticity and reliability on IHDS were also evaluated.Methods 200 patients with HIV infection were randomly selected from the minority ethnic groups in Guangxi.For each infected person,MMSE and IHDS blind scale were tested at the same period.Using the results from MMSE scale test as the gold standard,ROC curve and IHDS scale in Guangxi minority populations with different education levels which related to the diagnosis of dementia-HIV values were determined.The value of a specific sector under the IHDS sensitivity,specificity,and internal consistency coefficients was also evaluated.Results When considering the infected person did not differ on their educational level,the IHDS scale diagnostic cutoff appeared as 8.25,while 1HDS sensitivity as 0.925,specificity as 0.731 and Kappa as 0.477 (P＜0.001).When considering the extent of cultural differences did influence the prevalence of infection,the different education groups showed different IHDS diagnostic cutoff values.People with high school,secondary school or higher education levels,the IHDS diagnosis appeared to be 8.25,when sensitivity was 0.917,specificity was 0.895 and Kappa was 0.722 (P＜0.001).People with only primary education level,the IHDS appeared to be 7.25.When sensitivity was 0.875,specificity was 0.661 and Kappa was 0.372 (P＜0.001).Conclusion The IHDS diagnostic sector in Guangxi minority groups was lower than the internationally recommended level of diagnostic cutoff value (IHDS≤ 10 points).When using IHDS to perform the HIV related dementia screening program,in the minority areas of Guangxi,culture context,the degree and difference of HIV infection should be considered,especially in using IHDS diagnostic cutoff values.

Objective:To analyze the chemical constituents in Radix astragali by high-performance-liquid chromatography-time of flight mass spectrometry(HPLC-TOF/MS). Methods:An Agilent poroshell 120 SB-C18 column(100 mm × 3 mm,2. 7 μm)was adopt-ed. The mobile phase was composed of acetonitrile-0. 1% formic acid with nonlinear gradient elution. The flow rate was 0. 4 ml· min-1 . The UV detection wavelength was set at 254nm, the column temperature was 25℃ and the injection volume was 10μl. Electron spray ionization and positive mode was adopted, the flow and temperature of the carrier gas( N2 ) was 10 L·min-1 and 350℃, respec-tively. The capillary voltage was 4 kV, the bombardment voltage was 165 V, the spectra were recorded within the range of m/z 100~1 100. Results:A total of 24 chemical constituents were identified from Radix astragali by HPLC-TOF/MS simultaneously. Conclu-sion:An efficient and fast HPLC-TOF/MS approach has been established for studying the chemical constituents in Radix astragali, which lays the foundation for the study on pharmacodynamic material basis and quality control of Radix astragali.

In traditional oral practice, the presystemic interactions with gut microbiota is an important mechanism underlying the holistic health benefits of Chinese herbal medicines (CHMs), making the study of CHMs distinct from the research of Western medicines of which the systemic exposure (level in blood) is the starting point and the core. Gut microbial metabolism complements host metabolism in maintaining metabolic homeostasis of many biologically important endogenous molecules and the disposition of numerous exogenous compounds. Among them, the widely distributed gut bacterial β-glucuronidases (BGUSs) coordinate with host UDP-glucuronosyltransferases (UGTs) to play a role in the occurrence and intervention of diseases by affecting the glucuronidation homeostasis and altering the intestinal local and/or systemic exposure of endogenous compounds and xenobiotics. On one hand, many ingredients of CHMs undergo enterohepatic circulation; On the other hand, CHMs can act on BGUSs directly or indirectly change the distribution and function of BGUSs through reprogramming gut microbiome. The multiple interactions between BGUSs and CHMs may play an important role in the overall therapeutic benefits of CHMs. This work firstly summarizes the latest research progress on BGUSs; then the physiological, pathological and pharmacological significance of BGUSs are exemplified with representative endogenous and exogenous compounds from the aspects of nutrient utilization, metabolic homeostasis, and therapeutic response based on the varied substrate spectra of BGUSs; finally, the scattered data in literature were integrated to summarize the multiple interactions between BGUSs and CHMs, highlighting the important role of BGUSs in the holistic actions of CHMs.

Objective To investigate the ostengenie potential of adipose-derived stem cells(AD- SCs)when exposed to adenovirns containing hBMP-2 cDNA(Adv-hBMP-2)and offer a choice of cell source for gene therapy and tissue engineering.Methods Human adipose tissues were obtained from patients who received orthopaedic surgery or liposuction.ADSCs were obtained by digesting the adipose tissues.Firstly,flowcytometric analysis was performed for the confirmation of mesenchymal stem cell ori- gin and the surface markers including CD34,CD44,CD68,CD71,CD90,and CD105.The ADSCs were transfected by Adv-hBMP-2 and the effects were tested in vitro,lmmunoprecipitation and Western blotting and ELISA were performed for confirming BMP gone transduction and its stable expression.The transform of ADSCs was assessed by extracellular ALP staining,intracellular ALP spectrophotometry,von Kossa staining and RT-PCR.In the in vivo experiment ADSC-Adv-hBMP-2 cells were injected into the hind limb of nude mice and analyzed radiographically and histologically.Results ADSCs were successfully isolated from human adipose tissues.The isolated ADSCs expressed CD44,CD71,CD90 and CD105 and CD34 and CD68 were absent.The result confirmed the mesenchymal stem cell origin of the cells.West- ern blotting and ELISA confirmed successful and persistent hBMP-2 production by ADSC-Adv-hBMP-2 cells.Extracellular ALP staining,intracellular ALP spectrophotometry,yon Kossa staining and RT-PCR revealed that ADSCs treated with Adv-hBMP-2 had a tendency of transfering into osteoblast.X-ray and H&E sections from hind limb of nude mice injected with ADSC-Adv-hBMP-2 cells confirmed bone forma- tion at 2 weeks.Conclusions Liposuction aspirates contain abundant ADSCs that can be transduced with hBMP-2 gene,and the tranduced ADSCs differentiate into the osteoblast.ADSCs may be an ideal source of mesenchyme-lineage stem cells for gone therapy and tissue engineering.

Objective To evaluate the role of vascular endothelial growth factor-D(VEGF-D)in lymphangiogenesis of breast cancer,and investigate its relationship with some clinicopathological parameters and prognosis. Methods VEGF-D expression was detected in 85 cases with primary breast carcinoma by immunohistochemistry,and VEGF-D mRNA was detected by RT-PCR.Podoplanin monoclonal antibody was used to mark lymphatic endothelial cell and lymphatic vessel density(LVD) was counted.The relationship among the aboved parameters,clinicopathological parameters and prognosis was analysed. Results It was revealed by immunohistochemistry that VEGF-D expression was ranked by 5 levels: negative,n=5(5.88%);"+",n=17(20%);"++",n=34(40%);"+++",n=22(25.88%),and "++++",n=7(8.24%).VEGF-D expression was associated with tumor lymph node metastasis and TNM clinical stage(P

Objective To study absorption of shegan heji marker components in blood and their excretion in urine and feces of rats, after intragastric administration of shegan heji. Methods LC-MS/MS was used for determination of marker compounds. Rat metabolic cage technology was employed. Results Excretion of marker components were completed 24 hours after administration. Conclusion Ephedrine can be excreted from rats within 24 hours. The possibility of mutual transformation of flavonoids exists in the body. Taking shegan heji will not cause accumulation of ephedrine and flavonoids in the body.

BACKGROUNDOsteosarcoma (OS) is the most common primary malignant tumor of bone. Mouse models of human OS can invariably provide greater insight into the complex mechanisms that underlie the development and pathogenesis of this aggressive tumor. Bioluminescence technology favored tracing cancer cells in vivo. In this study, an OS model was described and evaluated using human OS cell line, Saos2, labeled with luciferase (Saos2-luc).

METHODSSaos2 cells were infected by lentivirus loading a firefly luciferase gene. Luciferase expression of Saos2-luc cells was characterized both in vitro and in vivo. Specific biologic and oncologic features of Saos2-luc cells were analyzed. The OS was established as orthotopic xenografts in nude mice. Both orthotopic tumors and spontaneous lung metastasis were analyzed.

RESULTSTumorigenesis and spontaneous lung metastasis in nude mice could be monitored in vivo through in vivo imaging system. The enhancement in proliferation, migration and invasion abilities and the attenuation in adhesion ability were observed in Saos2-luc cells compared with Saos2 cells. Furthermore, there were the up-regulation of Osteocalcin, CCR10, CXCR1 and ID1 and the down-regulation of ALP, collagen I, CCR1, CCR3, CXCR3, NID and N-cadherin in Saos2-luc cells compare to Saos2 cells. The rate of spontaneous lung metastasis in Saos2-luc cells was higher than that in Saos2 cells, although without significant difference.

CONCLUSIONSLentivirus transfection may cause alteration of gene expression profiles and further biological functions. This model can be used in the elucidation of molecular mechanisms of tumorigenesis and the screening of new therapeutic agents.

Animals ; Carcinogenesis ; pathology ; Cell Line, Tumor ; Humans ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Neoplasm Metastasis ; pathology ; Osteosarcoma ; pathology ; Real-Time Polymerase Chain Reaction ; Reverse Transcriptase Polymerase Chain Reaction ; Transplantation, Heterologous