SUMMARY Regenerationofbonetissue,aswellasothertissues,requiresinvolvementandinteraction of cells,scaffolds and relevant growth factors,among which growth factors play a crucial role in maintai-ning the stability of microenvironment.Nel-like-type 1 molecule (NELL-1 ),a novel growth factor in tis-sue engineering,has been studied intensively in recent years.Researches mainly covered gene and pro-tein structure and their expression profiling,biological function,molecular mechanisms and disease rele-vance.NELL-1 expressed in embryonic tissue is essential for growth and development of bone tissue. NELL-1 presents excellent abilities of inducing bone and cartilage regeneration,especially with high spe-cificity to chondrocyte lineage.Compared with classic osteogenic growth factor bone morphogenetic pro-tein 2 (BMP-2),the process of osteogenesis interacted with NELL-1 exhibits stronger specificity,higher bone density and fewerside effects.Furthermore,a recent study shows synergistic effects of NELL-1 and BMP-2.NELL-1 enhances the osteogenic reaction induced by BMP-2 of cells and notably declines in-flammation response caused by BMP-2.This review evaluates the current research progress of the function and application of NELL-1 by the systematic method of evidence-based medicine.

AIM:To investigate the expression of poly (ADP-ribose) polymerase-1 (PARP-1) in the epithelial ovarian cancer ( EOC) and its relationship with epithelial-mesenchymal transition ( EMT) .METHODS:The expression of PARP-1, E-cadherin, vimentin and Snail was detected in the EOC and benign ovarian tumor tissues by immunohistochemi -cal method and real-time PCR.The expression of PARP-1, E-cadherin, vimentin and Snail proteins in the SKOV3 cells treated with efficient PARP-1 inhibitor PJ34 was determined by Western blotting .RESULTS:The positive expression rates of PARP-1, vimentin and Snail were significantly higher in the EOC than that in the benign ovarian tumor tissues , whereas the positive expression rate of E-cadherin was the opposite (P<0.05).The expression of PARP-1, E-cadherin, vimentin and Snail in the EOC was associated with the histological grade , clinical stage and lymphatic metastasis (P<0.05), but no relationship with age and pathological types was observed .The expression of E-cadherin in the EOC was negatively co-related to that of PARP-1.In contrast, the expression of vimentin and Snail in the EOC was positively co-related to that of PARP-1.The relative mRNA expression of PARP-1, vimentin and Snail in the EOC was significantly higher than that in the benign ovarian tumor tissues ( P<0.05) , while the mRNA expression of E-cadherin in the EOC was remarkably lower than that in the benign ovarian tumor tissues (P<0.05).The protein expression of PARP-1, vimentin and Snail in the SKOV3 cells was significantly decreased (P<0.05), while E-cadherin protein was increased after treated with PJ 34(P<0.05). CONCLUSION:PARP-1 may contribute to the onset of EMT in the EOC by regulating the expression of E -cadherin, vim-entin and Snail.The role of PARP-1, which is relevant to EMT, might be important in the development of ovarian cancer .

The poor chemical stability and solubility, easy volatile and other shortcomings of TCM volatile oils can be improved after they are included by cyclodextrin. Therefore, cyclodextrin inclusion technology is widely used in the preparation process of volatile oil. This article reviewed the domain of cyclodextrin inclusion of volatile oils, the types of cyclodextrin, inclusion technologies, characterization for inclusion, components changes before and after inclusion, and dissolution characteristics of cyclodextrin inclusion.

Objective To investigate the factors affecting the stability of the volatile oil extracted fromYinqiaosan Decoction.Methods The main chemical compositions and the extraction repetitiveness of the compound volatile oil were determined by GC-MS, and the stability of multiple extracted volatile oil was studied. Absorbance of the compound volatile oil was used as the evaluation index, and the factors affecting the stability of the of the compound volatile oil were investigated, such as illumination, temperatures and pH values of volatile oil solution and metal ions.Results The results of the GC-MS chromatograph indicated that the main chemical compositions of the compound volatile oil extracted fromYinqiaosan Decoction twice were the same. The results of the stability of the volatile oil showed that the preservation temperature and illumination affected the stability of the volatile oil to a certain extent. The absorbance values of the compound volatile oil changed slowly when it was stored at a relatively low temperature (4℃) and shielded from light, and it was less stable when stored in normal temperature and under illumination. Meanwhile, the absorbance of the compound volatile oil changed quickly in acid or alkaline solutions and was in instability. The metal ions, such as Cu2+ and Fe3+, have chemical reactions with the compositions of the compound volatile oil and there was a big change in the UV-Vis spectrum of the compound volatile oils.Conclusion The compound volatile oil should be stored at a relatively low temperature (4℃) and shielded from light. At the same time, it should be stored avoiding acids, alkaline and the metal ions, such as Cu2+ and Fe3+, to guarantee its stability. This study provides a reference for the preservation conditions and the preparation conditions of the compound volatile oil extracted fromYinqiaosan Decoction.

Objective:To detect the activity of IgY against complex bacteria in pharynx and throat.Methods:Purified antigens against bacteria in pharynx and throat was used to immunize egglaid hens.The eggs from immunized hens were collected and abstract IgY from the yolks.The antibody activity of IgY was detected by SDS-PAGE electrophoresis and ELISA.Results:SDS-PAGE electrophoresis represented at least twelve ladders,and the titer of ELISA was 1∶512.Conclusion:IgY antibody was obtained in egg yolk after immunized hens with complex bacteria.The activity of IgY was detected.IgY showed stable to heat.

Objective To evaluate the efficacy of STERRAD (r) 100S (low temperature hydrogen peroxide gas plasma sterilization system) by comparing with formaldehyde steam. Methods From 2005.1 to 2005.3, totally sterilization of 2 760 pieces of operating tools were carried out with formaldehyde steam. From 2005.1 to 2005.3, totally sterilization of 4 950 pieces of operating tools in 180 circles were carried out with STERRAD (r) 100S. The instrument damage rate, operating infection rate and satisfaction rate of medical workers of 2 groups were compared. Results All data were in favor of the efficacy of STERRAD (r) 100S. Conclusions STERRAD (r) 100S is a safe, effective, low temperature, and cost-effective sterilization system..

Objective:To construct high level prokaryotic cell expression vector carrying human upstream regulatory element binding protein1 (hureb1) gene for producing GST hureb1 fusion protein and generating anti hureb1 antibody Methods: After the Xho I/Not I fragment from hureb1 open reading frame was recombined into the prokaryotic expression vector pGEX 4T 2, the vector was transformed into E coli BL21(DE3) strain and induced with IPTG to express the fusion protein, GST hureb1 The crudely isolated fusion protein was applied to immunize rabbit and mouse for generating anti hureb1 polyclonal antibody and monoclonal antibody respectively Results:The prokaryotic expression vector expressing GST hureb1 fusion protein was constructed and induced to produce high level GST hureb1 that was detected up to 33 45% of the total bacterial protein expressed The GST hureb1 fusion protein immunizing animals generated high titer and specific anti hureb1 antibodies Conclusion:Recombinant hureb1 protein and anti hureb1 antibodies can be used to analyze the biological functions of hureb1

ObjectiveTo investigate the relationship between metastasis-associated gene 1 ( MTA1 )expression and invasive and metastatic ability of cervical cancer cell. MethodsThree kinds of plasmids pcDNA3( control group), pcDNA3-MTA1 ( MTA1 group) and pSilencer3. 1-MTA1-siRNA ( MTA1-siRNAgroup) were transfected into human cervical cancer cell line CaSki cells. Reverse transcription (RT)-PCR and western blot were used to detected MTA1 mRNA and protein expressions. The effects of MTA1 expression on CaSki cell growth and proliferation, cell migration, adhesion and invasion, and cell cycles were tested by methyl thiazolyl tetrazolium (MTT), clone formation experiment, wound-healing assay, transwell assay, adhesion assay and flow cytometry, respectively. In animal experiment, three groups of cells were inoculated to BALB/c nude mouse subcutaneously to observe tumor formation ability. ResultsCompared with control group, MTA1 mRNA and protein were significantly overexpressed in MTA1 group, while MTA1-siRNA group showed lower MTA1 expression. Compared with control group, MTA1 group showed significantly accelerated cell growth; while MTA1-siRNA group showed decreased cell growth since the second day (P＜0. 05). Clone formation number in control, MTA1 and MTA1-siRNA group were 133 ±6, 169 ± 10 and 57 ±5,respectively. MTA1 group showed accelerated cell formation, while MTA1-siRNA group showed the reverse effect compared with that in control group(P ＜ 0. 05 ). At 24, 48 and 72 hours after wounding, the healing ability of MTA1-siRNA group significantly lagged behind that in the control group, while MTA1 group showed accelerated cell healing ability. The adhesion rate of control, MTA1 and MTA1-siRNA group were (69. 3 ± 3. 6) ％, ( 80. 4 ± 5. 6 ) ％ and ( 39. 2 ± 7.4 ) ％ separately at 90 minutes after cell seeding. In contrast with control group, MTA1 group promoted the adhesion of CaSki cell to matrigel matrix, while MTA1-siRNA group inhibited the adhesion process (P ＜0. 05 ). In the migration assay, the number of cells migrated to the bottom side of the membrane in control,MTA1 and MTA1-siRNA group were 153 ± 17,247 ± 38 and 82 ± 10, respectively. The number of cells in the invasion assay were 231 ± 19,354 ± 36 and 76 ± 7, respectively. Compared with the control group, MTA1 group significantly increased the migration and invasion ability, while MTA 1-siRNA group showed lower cell migration and invasion ability (P ＜ 0. 05 ). In cell cycle experiment, no significant differences of cell proportions including G1, S and G2 stage were found among three groups (P ＞ 0.05).In animal experiment, compared with control group,MTA1 group showed accelersted tumor formation and growth,whilethe MTA1-siRNA group showed the reverse effect ( P ＜ 0. 05 ). ConclusionsMTA1 may play its roles to promote cervical cancer cell invasion, migration, adhesion, as well as cell growth and colony formation, while RNA interference against MTA1 may decrease the malignant phenotypes. This study shows that it will be an effective beginning to explore metastasis mechanisms and cancer gene therapy strategy targeting MTA1 in cervical cancer.

Objective To investigate the expression and significance of elastin and fibulin-5 in anterior vaginal tissue of women with pelvic organ prolapse (POP). Methods Between November 2006 and June 2008, 68 patients with POP underwent surgical treatment in Shengjing Hospital of China Medical University were enrolled in this study, who were classified into 10 patients with grade Ⅰ , 21 patients with grade Ⅱ , 25 patients with grade Ⅲ and 12 patients with grade Ⅳ in accordance with pelvic organ prolapse quantitation (POP-Q). Meanwhile, 18 cases with early cervical cancer at stage of Ⅰ b were treated by total hysterectomy and bilateral salpingo-oophorectomy, their anterior vaginal tissues were selected as controls. Immunohistochemical staining was performed to detect the expression of elastin and fibulin-5. Results (1 ) Elastin and fibulin-5 were mainly expressed at extracellular matrix(ECM). (2)The positive rate of fibulin-5 expression in anterior vaginal wall were 5% (2/37 ) in grade Ⅲ/Ⅳ and 26% (8/31 ) in grade Ⅰ/Ⅱ POP patients, which reached statistical difference (P = 0. 035). However, no statistical different expression was found between postmenopausal (13%, 8/60) and non-menopausal patients (2/8), vaginal delivery ≤ 2 (19%, 5/27 ) and ＞ 2 patients (12%, 5/41, P ＞ 0. 05 ). (3 ) The positive rate of elastin expression in anterior vaginal wall in POP group was 31% (21/68), which was significantly lower than 72% (13/18) of control group (P = 0. 002). Among POP group, 19% (7/37) of elastin expression in grade Ⅲ/Ⅳ POP was significantly lower than 45% (14/31 ) in grade Ⅰ/Ⅱ of POP patients. However, no statistical difference was found between postmenopausal (30%, 18/60 ) and non-menopausal patients (3/8 ), vaginal delivery ≤ 2 (26%, 7/27) and ＞2 patients(34%, 14/41, P＞0. 05). (4)In POP group, both positive expression of fibulin-5 and elastin of anterior vaginal wall was in 6 cases, both negative expression of fibulin-5 and elastin was in 43 cases. It was illustrated that elastin and fibulin-5 had an positive relationship (P = 0. 031 ). Conclusion The decreased expression of elastin and fibolin-5 was correlated with degree of POP, which indicated that elastin and fibulin-5 may play a role in the pathogenesis of POP.

Objective To observe the effects of Radix Hedysari on collagen area, hyaluronic acid (HA) and laminin (LN) of lung tissues in rat with pulmonary interstitial fibrosis; To investigate the mechanism and screen the best active ingredients in Radix Hedysari.Methods Totally 144 SPF Wistar rats were randomly divided into control group, model group, metacortandracin group, and Radix Hedysari polysaccharide, flavone and saponin groups were set as high-, medium-, and low-dose groups. Pulmonary interstitial fibrosis models were set up by dropping bleomycin into air tube of rats. All groups were taken corresponding medicine with appropriate dose daily on day 2 after models were established for 28 days. Collagen fibrils in lung tissue were observed by Masson dying and contents of HA and LN in lung tissue of rats in each group were detected by radioimmunoassay.Results Compared with the model group, pulmonary alveoli in Radix Hedysari flavone high-, medium-, and low-dose groups and Radix Hedysari polysaccharide low-dose group were relieved obviously; collagenous fiber area in Radix Hedysari flavone low-dose group, Radix Hedysari polysaccharide medium-dose group, and Radix Hedysari saponin low-dose group decreased obviously. Compared with the model group, the content of HA in Radix Hedysari polysaccharide high-dose group, Radix Hedysari flavone high-, medium-, and low-dose groups, and Radix Hedysari saponin high-dose group decreased obviously; the content of LN in Radix Hedysari polysaccharide high-dose group, Radix Hedysari flavone medium- and low-dose groups, and Radix Hedysari saponin medium- and low-dose groups decreased obviously. Conclusion Polysaccharide, flavone and saponin of Radix Hedysari have the abilities to alleviate inflammation of pulmonary alveoli, inhibit proliferation and deposition of collagen fibrils, and reduce the contents of HA and LA in lung tissue. Among these active ingredients, flavone has the best effect.