1.NF-?B site of promotor mediates nicotine-induced ICAM-1 expression in human umbilical vein endothelial cells
Chinese Journal of Pathophysiology 1986;0(02):-
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
2.Preparation and bone repair effect of platelet-rich gel
Meng LU ; Weihui CHEN ; Yanzheng QU
Chinese Journal of Tissue Engineering Research 2007;0(23):-
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.
3.Establishment of a cell line of Tca8113 with stable expression of nm23-H1 by gene transfection
Keqian ZHI ; Weihui CHEN ; Yuming WEN
Journal of Practical Stomatology 2000;0(06):-
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.
4.The study of increased sensitivity to cisplatin by nm23-H1 in vitro
Keqian ZHI ; Weihui CHEN ; Yuming WEN ; Zhuangqun YANG
Journal of Xi'an Jiaotong University(Medical Sciences) 1981;0(02):-
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.
5.THE DISTRIBUTION OF BACILLUS THURINGIENSIS AND CRY GENE DIVERSITY IN CHINA
Jinhong WANG ; Weihui WU ; Yuehua CHEN ; Gaixin REN ;
Microbiology 1992;0(03):-
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.
6.Occlusal stimulation regulates bone remodeling during bone defect repair using collagen substitutes:a morphological analysis
Chuanqing MAO ; Jin WANG ; Chengyong WANG ; Meng LU ; Weihui CHEN
Chinese Journal of Tissue Engineering Research 2015;(47):7603-7607
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.
7.Anti-mullerian hormone in seminal plasma and serum:association with sperm parameters
Jiajie LIU ; Zuchang WU ; Xiaowu FANG ; Jianhong WEI ; Weihui CHEN
International Journal of Laboratory Medicine 2017;38(2):226-228
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.
8.Correlation of innervation to osseointegration and osseoperception of implants
Chengyong WANG ; Weihui CHEN ; Hai LIN ; Yanzheng QU
Chinese Journal of Tissue Engineering Research 2010;14(4):684-688
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.
9.Significance of the expression of microRNA in the peripheral blood of patients with acute pancreatitis
Gaofeng ZHANG ; Weihui LIU ; Yi WEN ; Wei YANG ; Tao CHEN ; Lijun TANG
Chinese Journal of Digestive Surgery 2014;13(11):875-879
Objective To predict the severity of acute pancreatitis by detecting the changes of microRNA in the serum,and whether paracentesis catheter drainage (PCD) should be applied to the patients.Methods The peripheral blood of 120 patients with acute pancreatitis who were admitted to the General Hospital of Chengdu Military Command from October 2013 to March 2014 were collected.Thirty-five patients with severe acute pancreatitis (SAP) or moderately severe acute pancreatitis (MSAP) were in the group A,and 85 patients with mild acute pancreatitis (MAP) were in the group B.The scores of the acute physiology and chronic health evaluation Ⅱ (APACHE Ⅱ),RANSON and bedside index for severity in acute pancreatitis (BISAP) were assessed.The expressions of the miR-146a,miR-10b,miR-21 and miR-26a in the serum were detected by the real-time quantitative PCR.The differences in the expressions of the 4 kinds of microRNAs in the 2 groups were compared,and the correlation between the 3 exaluation system was analyzed.Factors influencing the timing of the PCD for the PCD patients were analyzed.The measurement data were analyzed using the t test,and the relationship between the variables was analyzed using the linear correlation analysis or the rank correlation analysis.Univariate or multivariate analysis was done by the Logistic regression analysis.Results (1) The scores of the APACHE Ⅱ,RANSON and BISAP were 8.28 ±0.61,3.42 ±0.54 and 1.71 ±0.32 in the group A,and 3.18 ±0.52,1.43 ±0.25 and 0.37 ±0.06 in the group B,with significant differences between the 2 groups (t =4.266,7.809,4.113,P < 0.05).(2) The expressions of the miR-146a,miR-10b,miR-21 and miR-26a were 1.41 ± 0.21,2.94 ± 0.49,1.62 ± 0.25,1.21 ± 0.20 in the group A,and 6.29 ± 0.91,0.52 ± 0.09,2.82 ± 0.33 and 3.57 ± 0.64 in the group B.There were significant differences in the expression of the miR-146a and miR-10b between the 2 groups (t=-2.156,2.110,P <0.05),while no significant difference in the miR-21,miR-26a was detected between the 2 groups (t =-1.114,-1.571,P > 0.05).(3) There was correlation between the expressions of the miR-146a,miR-10b and the APACHE Ⅱ,RANSON,BISAP in the group A (r =-0.826,0.837,-0.874,0.866,-0.833,0.899,P < 0.05),while no correlation was detected between the expressions of the miR-21,miR-26a and the 3 exaluation systems in the group A (r =0.642,0.321,0.701,0.750,0.716,0.716,P > 0.05).There was no significant difference between the miR-146a,miR-10b,miR-21,miR-26a and the APACHE Ⅱ (r =0.067,0.347,0.133,0.111,P>0.05),RANSON (r=0.178,0.078,0.092,0.142,P >0.05) and BISAP (r =0.103,0.260,0.216,0.285,P > 0.05) in the group B.(4) The results of univariate analysis showed that miR-10b,RANSON and BISAP were the factors influencing the timing of PCD (OR =4.170,5.612,2.500,95 % confidence interval:1.092-15.932,1.232-21.622,1.190-5.254,P < 0.05).The results of multivariate analysis showed that miR-10b was the factor influencing the timing of PCD (OR =2.374,95% confidence interval:1.115-5.056,P < 0.05).Conclusions miR-10b and miR-146a might be the predictors of severity of severe acute pancreatitis; miR-10b might be the indicator in judging whether PCD should be applied.
10.Denervation of the inferior alveolar nerve is associated with bone formation and bone mass maintenance in extraction sockets
Chengyong WANG ; Weihui CHEN ; Meng LU ; Jin WANG ; Chuanqing MAO ; Hai LIN
Chinese Journal of Tissue Engineering Research 2015;(42):6740-6745
BACKGROUND: There is a close relationship between innervations and bone formation and bone mass maintenance in the extraction sockets. OBJECTIVE:To study the possible effect of denervations on the regulation of new bone formation and bone mass maintenance in the extraction sockets. METHODS:The unilateral inferior nerve of dogs was sectioned to establish an animal denervation model. The normal side was used as control. After model establishment, the premolars of denervated side and normal side were extracted. Histological method was used to test new bone formation and bone mass maintenance in the extraction sockets at the 2nd, 4th, 8th and 12th weeks after tooth extraction. RESULTS AND CONCLUSION:The percentage of new bone areas in the extraction sockets was significantly lower in the experiment group than the control group at weeks 2, 4, 8 after tooth extraction (P < 0.01). The height difference between the buccal and lingual alveolar ridge was higher in the experimental group than the control group at weeks 2, 4, 8, 12 after tooth extraction (P < 0.05 or P < 0.01). These findings indicate that denervation is closely related with new bone formation and bone mass maintenance in the extraction sockets.