Cloning and Expression of Rat Transforming Growth Factor β1 cDNA in Osteoblasts

In order to study transforming growth factor81 (TGF51) in molecular level and investigate thefeasibility of TGF51 gene therapy for bone defects,an eukaryotic expression vector containing ratTGF51 cDNA was constructed. The expression ofTGF51 in the transfected osteoblasts was observed.1 MATERIALS AND METHODS1. 1 Animals and Cell CultureSprague--Dawley rats, I adult and 5 newborn,were purchased from Animal Center of Tong nMedical University. Lymphocytes, isolated fromperipheral…     

Effects of Icariin on Expression of OPN mRNA and Type ⅠCollagen in Rat Osteoblasts in Vitro

Osteopontin(OPN)isagroupofactivepro teinswhichareproducedbybonetissue.OPNcanregulatethelocalfunctionandplayanimportantroleinconstructionandremodelingofbone.typeⅠcollagenisthemaincomponentinbonematrixwhichplaysroleinbonerestoration.Osteoporosisiscausedbyboneresorptionmorethanbonefor mationandosteoblastsarethefunctionalcellswhichareresponsibleforboneformation.Thisstudywasdesignedtoshowthemolecularmecha nismsofIcariinanditseffectonexpressionofOPNmRNAandtypeⅠcollageninratosteoblastsinvitroa…     

Effects of Tripterine on mRNA Expression of Oncogene c-myc and Platelet-Derived Growth Factor of Vascular Smooth Muscle Cells in Rats

Percutaneoustransluminalcoronaryangioplasty(PTCA)hasbecomeacommonlyemployedtechniqueinthetreatmentofcoronaryheartdiseaseoverthelastdecade.However,thefrequentoccurrenceofrestenosisfollowing30%to40%ofinitiallysuccessfulprocedureremainsthemajorlimitationofthetechnique(').Studiessuggestthatoverproliferationofvascularsmoothmusclecells(VSMCs),playingapredominantroleintherestenoticprocess,inwhichobviouslytheactivationofnuclearoncogenes(c--myc,etc)andincreaseofgrowthfactorsreleasedwithinthearterial…     

Effects of Erigeron Injection on Vascular Endothelium in Patients

Objective： To observe the levels of serum C-reactive protein （CRP）, endothelin-1 （ET-1）, nitric oxide （NO）, and superoxide dismutase （SOD） in patients after intracoronary stenting （ICS）, and the effects of Erigeron Injection （E1） on them. Methods： Seventy-two patients, who received ICS and had symptoms of chest stuffiness, palpitation and chest pain, were randomly divided into two groups, with 36 patients in the control group treated with Plavix alone for anti-platelet aggregation, and the other 36 patients in the treated group treated with Plavix and El in combination. CRP, ET-1, NO and SOD were determined and compared before and 1, 2 and 3 weeks after treatment. Results： As compared with those in the control group, improvement of symptoms in the treated group was significantly better, with the levels of CRP and ET-1 lower and levels of SOD and NO higher or approaching to normal ranges and significant difference was shown between the two groups （P〈0.01）. Conclusion： El could alleviate uncomfortable feelings such as chest stuffiness in patients after lOS, and improve the function of vascular endothelium.     

The Effect of Hypoxia on Expression of Basic Fibroblast Growth Factor in Pulmonary Vascular Pericytes

Recentstudiessuggestedthathypoxiacouldin-ducepulmonaryvascularpericytes(PC)proliferationanddiffeTentiationintosmoothmuscle--likecells,whichleadtomuscularizationofnon--muscularartieriolesofthelungduringthedevelopmentofhy-poxicpulmonaryhypertension['-'].However,themechanismofPCproliferationanddifferentiationre-mainsunclear.Itisknownthatbasicfibroblastgrowthfactor(bFGF)isanimportantfactorthatregulatescellproliferationanddifferentiation,andre-centlymoreimportancewasattachedintothestudyofvascula…     

Study of Rat Osteoblasts Transfected by Transforming Growth Factor β1 Gene

Summary: In order to investigate the effect of TGFβ1 gene transfer on the biological characteristics,the effects of gene transfer and supernatant of transfected osteoblasts on the proliferation and ALP activity of osteoblasts were detected by 3H-TdR and MTT. Our results showed that TGFβ1 gene transfer had no effect on the biological characteristics and the activated supernatant of transfected os-teoblasts stimulated proliferation and inhibited ALP activity of osteoblasts. TGFβ1 gene transfer could promote the expression of TGFβ1 and the biological characteristics of transfected osteoblasts were sta-ble, which might be helpful for gene therapy of bone defects in vivo.     

Modulation of Isoflavones on Bone—nodule Formation in Rat Calvaria Osteoblasts in vitro

Objective:to observe the effects of two main isoflavones,daidzein and genistein on the bone-nodule formation in rat calvaria osteoblasts in vitro.Methods:Osteoblasts obtained from newborn Sprague-dawley rat calvarias were cultured for several generations.The second generation cells were cultured in Minimum Essential Medium supplemenmted with ascorbic acid and Na-beta-glycerophosphate for several days,in the presence of daidzein and genistein,with or without the estrogen receptor antagonist ICI 182780.Number of nodules was counted at the end of the incubation period(day 20) by staining with Alizarin Red S calcium stain.The release of osteocalcin,as a marker of osteoblast activity,was also determined on day 7 and 12 during the incubation period.Results:compared with the control,the numbers of nodules were both increased by incubation with daidzin and genistein,17β-estradiol was used as a positive control and proved to be a more effective inducer of the increase in bone-nodules formation than daidzein and genisterin.The release of osteocalcin into culture media was also increased in the presence of daidzein and genistein,as well as 17β-estradiol on day 7 and day 12(day 12 were higher).The estrogen receptor antagonist ICI 182780 completely blocked the genistein-and 17β0estradiol-induced increase of nodule numbers and osteocalcin release in osteoblasts.Howerver,the effects induced by daidzein could not be inhibited by ICI 182780.Conclusion:These findings suggest that geinistein can stimulate bone-nodule formation and increase the release of osteocalcin in rat osteoblasts.The effects,like those induced by 17β-estradiol,are mediated by the estrogen receptor dependent pathway,Daidzin also can stimulate bone-nodule formation and increase the release of osteocalcin in rat osteoblasts,but it is not,at least not merely,mediated by the estrogen receptor dependent pathway.     

Effects of Erigeron Injection on Vascular Endothelium in Patients after Intracoronary Stenting

Objective: To observe the levels of serum C-reactive protein (CRP), endothelin-1 (ET-1), nitric oxide (NO), and superoxide dismutase (SOD) in patients after intracoronary stenting (ICS), and the effects of Erigeron Injection (El) on them. Methods:Seventy-two patients, who received ICS and had symptoms of chest stuffiness, palpitation and chest pain, were randomly divided into two groups, with 36 patients in the control group treated with Plavix alone for anti-platelet aggregation, and the other 36 patients in the treated group treated with Plavix and El in combination. CRP, ET-1, NO and SOD were determined and compared before and 1, 2 and 3 weeks after treatment. Results: As compared with those in the control group, improvement of symptoms in the treated group was significantly better, with the levels of CRP and ET-1 lower and levels of SOD and NO higher or approaching to normal ranges and significant difference was shown between the two groups (P<0.01). Conclusion: El could alleviate uncomfortable feelings such as chest stuffiness in patients after ICS, and improve the function of vascular endothelium.     

Regulation of Hypoxic Response Elements on the Expression of Vascular Endothelial Growth Factor Gene Transfected to Rat Skeletal Myoblasts under Hypoxic Environment

The regulation of hypoxic response elements on the expression of vascular endothelial growth factor （VEGF） gene transfected to primary cultured rat skeletal myoblasts under hypoxic environment was investigated. pEGFP-C3-9HRE-CMV-VEGF vector was constructed with molecular biology technique and transfected to primary cultured rat skeletal myoblasts by lipofectamine in vitro. Gene expression of transfected myoblasts was detected by RT-PCR, Western blot and fluorescence microscope under different oxygen concentrations and different hypoxia time. The results showed that in hypoxia group, the VEGF gene bands were seen and with the decrease of oxygen concentrations and prolongation of hypoxia time, the expression of VEGF mRNA was obviously increased. Under hypoxic environment, the expression of VEGF protein in the transfected myoblasts was significantly increased. EGFP was expressed only under hypoxic environment but not under normoxic environment. It was concluded that hypoxia promoter could be constructed with HRE and regulate the expression of VEGF gene under hypoxic and normoxic environment, which could enhance the re- liability of gene therapy.     

Effect of Kang''''ai Injection(康艾注射液) on Serum Level of Soluble Interleukin-2 Receptor and Vascular Endothelial Growth Factor in Patients with Esophageal Carcinoma during Radiotherapy

Radiotherapy(RT)is an important ap-proach for treatment of esophageal carcinoma(EC),especially for patients who have missedthe chance of surgical section.The efficacy ofRTis not satisfactory for its1-year,3-year and5-year survival rates being merely50%,20%,and10%respectively(1).Researches in recent years showed thatbetter efficacy can be obtained by combiningRTwith Chinese herbal medicines for EC thatincapable to be resected.Kang ai Injection(康艾注射液,KAI)is a Chinese herbal prep-arati…     

Expression of Vascular Endothelial Growth Factor in Hepatocellular Carcinoma and Its Relationship to Tumor Growth and Metastasis

Vascularendothelialgrowthfactor(VEGF)isanewlydiscoveredgrowthfactorwhichmayinducetumorangiogenesisll].Inthepresentstudy,immunohistochemistryandNorthernblotwereusedtoinvestigatetheVEGFexpressioninhepatocellularcarcilloma(HCC)withtheattempttounderstanditsrelationshiptothemetastasls.IMATERIALSANDMETHODS1.1TissueSamplesThespecimensinvestigatedwereobtainedfromZII)atientswithHCCoperatedattheaffiliatedhospitalsofouruniversity.Nineofthepatientsshowedclinicallymetastasiswithportalveintu…     

Dynamic Expression of Tumor Necrosis Factor-α and Vascular Endothelial Growth Factor in Rat Model of Pulmonary Emphysema Induced by Smoke Exposure

In order to explore the roles of tumor necrosis factor-α(TNF-α) and vascular endothelial growth factor(VEGF) in the pathogenesis of pulmonary emphysema,male Wistar rats were randomized into group A1,group A2.5 and group A4,each with smoke exposure for 1 month,2.5 months or 4 months,respectively.Group B1,group B2.5 and group B4 were used as non smoking controls at corresponding time points.TNF-α in bronchoalveolar lavage fluid(BALF) and expression of VEGF in lung tissue was determined by ELISA or by SABC immunohistochemistry assay either.Lung slices were stained with hematoxylin and eosin(HE).Results showed that in animal with smoke exposure the mean linear interceptor(Lm),an index of pulmonary emphysema and the content of TNF-α in BALF increased gradually,on contrary,the expression of VEGF in lung tissue decreased(P<0.05).This phenomenon was not obvious in animals without smoke exposure.Lm was negatively correlated to the VEGF expression(γ=-0.81,P<0.01) and positively correlated to TNF-α concentration(γ = 0.52,P<0.004),which implies that smoke exposure decreased the expression of VEGF and increased the expression of TNF-α.It is plausible to speculate that the imbalance of TNF-α and VEGF may play an important role in the pathogenesis of smoke-induced pulmonary emphysema.     

Effects of Ureaplasma Urealyticum on Fas Ligand Expression in Rat Sertoli Cell

Testis is an important male sexual organ. Sertoli cell, which is the main stromacell in testis, provides the microenvironment for the differentiation and maturation ofspermatogenic cell. It can also form a blood--testis barrier to protect the spermatogenic cells and secrete several materials that can affect other cells and itself. Sertolicell also secretes many kinds of cytokine, which play important roles in sustainingand regulating the testis microenvironment and reproductive function. In re…     

Effects of Danshen Injection on the Malignant Obstructive Jaundice in the SD Rat Model

A malignant obstructive jaundice model[1] was es-tablished in rats by inoculation of transplanted tumor (inoculated with the Walker-256 hepatocarcine line) into the hepatic portal of SD mouse to cause obstruction of the biliary tract by the infiltration and metastasis of hepatocellular carcinoma. The effect of Danshen on the growth of hepatocellular carcinoma in the malignant ob-struction jaundice in the SD rat model was studied and the effect of Danshen was compared with other agents in ter…     

Intrastriatal Gene Transfer of Vascular Endothelial Growth Factor Rescues Dopaminergic Neurons in a Rat Parkinson's Disease Model

Parkinson's disease (PD) is a chronic progressivegerontic disease, which is characterized by the degeneration of dopaminergic neurons in substantia nigra. Efforhas been made to find effective treatment to arrest oslow down the degeneration of dopaminergic neuronsOnce regarded as an endothelial cell-specific factorvascular endothelial growth factor (VEGF), in recenyears, was unexpectedly found to be a critical player inneurodegeneration, and have direct neurotrophic andneuroprotective effects…     

Effects of Vascular Endothelial Cell Growth Factor on Fibrovascular Ingrowth into Rabbit＇s Hydroxyapatite Orbital Implant

The effects of different concentrations of vascular endothelial cell growth factor (VEGF) on the fibrovascular ingrowth into rabbits hydroxyapatite orbital implant were investigated. Twelve New Zealand white i‘abbits were divided into 3 groups and received hydroxyapatite orbital implant surgery in their right eyes. Before and after the operation, the implants were treated with I0 ng/ml VEGF, 100 ng/ml VEGF, or normal saline as control group. The animals received technetium bones scan at 2, 4, and 6 weeks postoperatively. The mean radioactivity counts within region of interest (ROI) of the surgery eye (R) and the non-surgery eye (L) in the same animal were tested,and the R/L ratios were calculated. The implants were harvested at 6th weeks and examined histopathologically. The results showed that at second week, there was no significant difference in mean R/L ratios between VEGF group and control group (F=2.83, P=0. 111); At 4th week (F=7. 728, P=0. 011) and 6th week (F=7. 831, P=0. 011) postoperatively, the mean ratios in VEGF groups were significantly higher than that in control group. At 6th week postoperatively,the fibrovascularization rates in VEGF groups were higher than in control group significantly (F=8. 711, P=0. 008). It was suggested that VEGF could promote the fibrovascular ingrowth into hydroxyapatite orbital implant, thus might shorten the time required for complete vascularization of the HA orbital implant.