AIM: To study the mechanisms of nicotine-induced expression of intercellular adhesion molecule-1(ICAM-1). METHODS: Related luciferase reporter gene plasmids were constructed with molecular cloning techniques;above plasmids and intracontrol plasmid pSV-?-gal were co-transfected into human umbilical vein endothelial cells(HUVECs) with eukaryotic gene transfection techniques; the relative luciferase activities were detected in the transfected HUVECs. RESULTS: Series of luciferase reporter gene containing different sequences of human ICAM-1 promotor and site-directed mutants of NF-?B and Sp-1 in promotor were successfully constructed; Nicotine could increase the expression of luciferase reporter gene plasmid containing-579 bp(pGL3E-579/+36),-230 bp(pGL3E-230/+36) and mutated Sp-1 version(pGL3E-Sp-1-MU)( P

Platelet-rich plasma is a volume of autogenous plasma with a high platelet concentration. Platelet-rich gel which is polymerized from platelet-rich plasma becomes a hot topic in the repairing of jaw bones in oral and dentofacial surgery. Presently, the mechanism of platelet-rich gel in enhancement of bone defect repair has not been clarified. Generally, we believed that the unifiation of blood platelet degranulation released many high-concentration growth factor during polymerization stimulated bone formation and accelerated bone repair. In addition, the application of platelet-rich gel has some problems. Thus, wide application of platelet-rich gel in tissue engineering deserves further studies. With the development, platelet-rich gel should be widely used in bone tissue engineering.

Objective:To establish a tongue cancer cell line that stably overexpresses nm23-H1.Methods:The reconstructed plasmid, pCMV-BamH1-Neo-nm23-H1, was transfected into bacteria and amplified. The plasmid was identified by electrophoresis on a 10 g/L agarose gel and stained with ethidium bromide and then transfected to the cell line of Tca8113 by lipofectin strategy and was selected by G418 for 5 weeks. The stable expression of nm23-H1 was identified by immunofluorescence,Western-blotting and flow cytometer.Results:Restriction endonuclease Bam H1 examination showed that 986 bp of nm23-H1 was in pCMV-Bam-Neo vector. 5 weeks after transfection positive clones were obtained and the transfected cells were proliferated to confluent.Immunohistochemical examination,Western blot and flow cytometry showed that nm23-H1 expression was increased in the transfected cells.Conclusion:A tongue cancer cell line expressing nm23-H1 is established.

BACKGROUND: Consummate innervation plays an important role in bone formation. Nerve injury can impact normal bone metabolism, whereas bone regeneration depends on nerve regeneration in the innervation site to some degrees. Modified implant surface structure or usage of growth factors in local region can promote osseointegretion.OBJECTIVE: To retrospectively analyze effects of nervous system dominance bone metabolism and denervation on bone remodeling, innervated establishment and osseoperception during osseointegration following implantation.METHODS: The first author retrieved Pubmed database (http://www.ncbi.nlm.nih.gov/PubMed) and Wanfang database (http://www.wanfangdata.com.cn) (1990/2008) for publications of effects of nervous system dominance bone metabolism and denervation on bone remodeling, innervated establishment and osseoperception during osseointegration following implantation,with the key words of "bone metabolism, innervation, osseointegrationosseoperception" . Articles of repetitive contents were excluded. A total of 54 literatures were primarily obtained. According to inclusion criteria, 28 literatures were selected for summary.RESULTS AND CONCLUSION: Signals derived from the nervous system exert potent effects on osteoclast and osteoblast function. A ubiquitous innervation of all periosteal surfaces exists and its disruption affects bone remodeling. Several neuropeptides, neurohormones, nerotransmitters and their receptors are detectable in bone, and make it clear that nervous system is involved in bone metabolism and regeneration. Establishment of innervation of soft and hard tissue around implants may play an important role in osseointegration and osseoperception. Further studies will be required to explore the basic of anatomy,neurophysiology and neuropathology, which forms osseoperception.

Objective To study the mechanism of increased sensitivity to cisplatin by nm23-H1.Methods The plasmid,pCMV-BamH1-Neo-nm23-H1,was transfected to cell line Tca8113 by lipofection strategy and was selected by G418 for 5 weeks.The stable products of nm23-H1 gene were identified by Immunohistofluorescence and Western-blotting.Using MTT and flow cytometer,we detected the changes of cell mortality rate,apoptosis and mitochondrial membrane potential.Results We successfully established the cell line of stable overexpression of nm23-H1.In vitro,the cell mortality rate and apoptosis were increased significantly in stable expression cell line of nm23-H1,compared with those in Tca8113 cell line of non-transfection;this effect could be inhibited by oubain which is an inhibitor of Na(+)/K(+)-ATPase.Mitochondrial membrane potential was decreased significantly in stable expression cell line of nm23-H1.Conclusion Nm23-H1 can increase the sensitivity of cisplatin on Tca8113 cell line;The mechanism may be the mitochondrial membrane potential is decreased by nm23-H1 so that intracellular platinum are increased to finally cause the apoptosis and necrosis of cells.

Objective To explore the association of anti-mullerian hormone(AMH)in seminal plasma and serum with sperm counts and energy for male.Methods For 215 cases of healthy male selected from our reproductive clinic,with women′s reason for infertility,seminal plasma and serum AMH were detected,as semen parameters(sperm density,living rate,vitality and malformation rate),6 items of serum sex hormone.In seminal plasma and serum AMH respectively as the dependent variable,using multiple line-ar regression model to explore its quantitative relation with semen parameters and sex hormone levels.Results 215 cases were en-rolled,aged 34.28±5.70 years,while the median of the seminal plasma AMH was 0.47,quartile 0.05-3.09 pmol/ejaculation.The median of the serum AMH was 53.07,quartile 32.32 -72.20 pmol/L.Through multiple linear regression analysis,after adjusted by age and BMI,the seminal plasma of AMH and total number of sperm,sperm concentration,dynamic motility,total sperm activi-ty,serum inhibin B were positively correlated(P <0.05);The correlation between sperm morphology and other serum sex hormone had no statistical significance(P >0.05);Serum AMH negatively correlated with serum FSH,with serum inhibin B positively(P <0.05);Seminal plasma in various parameters and other related serum sex hormone had no statistical significance(P >0.05).Conclu-sion The seminal plasma of AMH were positively correlated with sperm concentration,sperm counts,sperm vitality,with the asso-ciation for serum AMH not yet found.

cryⅢ gene,they were found in 75 6%,67 9%,58 4% and14 5% of the strains respectively,no cryⅠ Ⅴ gene was found,10cry gene combination types were concluded.The Bt isolates which contained cryⅠ genes were further characterized by additional PCR detection with specific primers of the cryⅠAc,cryⅠC and cryⅠE genes.20 Bt isolates which contained cryⅠAc,cryⅠC,cryⅡ and cryⅤ genes were found,among them the strain Bt\|15A3 is high toxic to Heliothis armigera,Spodoptera exigua and Plutella xylostella,and has potential developing and applying value.

BACKGROUND:Occlusal stimulation is essential for mandible function and remodeling, but there is stil a lack of clear understanding about the effect of occlusal stimulation on the bone remodeling in the process of bone defect repair using bone grafts. OBJECTIVE:To analyze the possible regulative effect of occlusal stimulation on bone remodeling in the process of bone defect repair using colagen substitutes. METHODS:Standard models of bone defects were respectively established in left mandible and parietal bone area of adult Sprague-Dawley rats. Then the bone defects area were filed with colagen and bone meal. The differences of two bone defects areas were observed by X-ray, hematoxylin-eosin staining, Gomori staining, tartrate-resistant acid phosphatase staining and bone morphogenetic protein 2 immunohistochemical staining at the 12th week after operation. RESULTS AND CONCLUSION: New bone formation was visible in the bone defect regions of the mandible and parietal bone. The amount of lamelar bone formation and the degree of mineralization of the new bone were significantly increased in the parietal bone defect compared with the mandibular bone defect area, indicating the bone remodeling in the parietal bone defect area was better than that in the mandible bone defect area. The integral absorbance values of tartrate-resistant acid phosphatase and bone morphogenetic protein 2 in the parietal bone defect area were lower than those in the mandibular bone defect area, indicating that the viabilities of osteoblasts and osteoclasts in the parietal bone defect area were lower than those in the mandible bone defect area. These results demonstrate that occlusal stimulation may delay the bone remodeling during the repair of mandibular bone defects by regulating bone mineralization and maturation.

AIM: To assess the clinical efficacy of phacoemulsification, intraocular lens implantation with goniosynechialysis(PEI+GSL)for acute angle closure glaucoma(AACG)and cataract with extensive angle closure synechiae.

METHODS: A retrospective study, we studied 35 eyes of 32 patients with AACG and cataract in our hospital. The extent of anterior chamber angle-closure synechiae was defined as an eye with >180°. All patients underwent PEI+GSL and completed an ophthalmologic examination including vision, intraocular pressure(IOP), anterior chamber depth(ACD), angle-opening distance(AOD500), trabecular-iris space area(TISA500)were observed at 1d, 1wk, 1mo and 3mo after cataract surgery. The angle closure range and retinal nerve fiber layer(RNFL)thickness changes at postoperative 1mo and 3mo were observed, and recorded complications.

RESULTS: Postoperative 3mo BCVA(0.334±0.154)and IOP(14.63±3.59mmHg)were improved compared with preoperative(0.914±0.290, 42.54±8.06mmHg)(P<0.05). ACD(3.203±0.214mm), OCT angle parameters AOD500 and TISA500(0.308±0.014, 0.315±0.015mm, 0.134±0.013, 0.139±0.018mm2)were significantly increased compared with preoperation. The extent of angle closure with gonioscopy(72.32±28.33°)decreased compared preoperation(215.29°±30.66°), and RNFL thickness thinner than preoperation(P<0.001). Changes in AOD500 and TISA500 for both nasal and temporal were negatively correlated with IOP, but not with changes in ACD, and no significant complications occurred in the 3mo after surgery.

CONCLUSION: The treatment of PEI+GSL can improve vision, deeper ACD and effectively open ACA in the early stage, thus controlling IOP.

Objective To investigate the mechanism underlying the WNK4 kinasemediated inhibitory effect on BK channel. Methods Cos-7 cells were cotransfected with BK in combination with either CD4 (control group) or wild type WNK4 (WNK4-WT).Immunostaining and confocal microscopy,chemiluminescence,Western blotting analysis were then employed to determine the BK localization in cells,BK surface expression and total protein level,respectively.To further investigate whether the reduction of BK protein expression is due to an increase in degradation through a lysosomal pathway,BK protein level was determined after treated with bafilomycin A1(Baf A1),a proton pump inhibitor affecting lysosomal degradation. Results Immunostaining and confocal microscopic study showed that BK was localized both in plasma membrane and cytosol in the control group.After cells transfected with WNK4-WT,BK expression was markedly reduced.Chemiluminescent assay found that BK surface expression level was 299.9±18.6 in the control group,whereas it was significantly reduced (148.4±13.7,P＜0.01) in the WNK4-WT group.Western blotting analysis showed that total BK protein level was markedly reduced in the presence of WNK4-WT compared to the control group.WNK4-WT was shown to significantly reduce the BK total protein level (42.3％±15.2％) compared to the control group (100％) (P＜0.01).When the cells was treated with Bafilomycin A1 (Baf A1,0.5 μmol/L),WNK4-mediated reduction in BK protein was reversed (82.2％±12.1％,P＜0.05). Conclusions WNK4 inhibits total and surface protein expression of BK in Cos-7 cells whick is likely due to an increase in BK degradation through a lysosomal pathway.