OBJECTIVE: To study the effect of simvastatin on the expression of bone morphogenetic protein-2 (BMP-2) and alkaline phosphates (ALP) activity in the primary cultured bone marrow stromal cells, and to elucidate the mechanism of the anabolic osteogenetic effect of simvastatin. METHODS: Bone marrow stromal cells in femur and tibia of adult mouse were cultured in vitro. after treated with different concentrations of simvastatin (0, 0.1, 0.2, 0.5 and 1.0 mumol/L) or recombinant human BMP-2 for 72 hours, ALP activity of bone marrow stromal cells was determined. BMP-2 expression of bone marrow stromal cells was analyzed by using immunocytochemistry and Western blotting. RESULTS: After treated with simvastatin for 72 hours, BMP-2 expression increased, while little BMP-2 expression could be observed in the control group. ALP activity also increased in a dose-dependent manner; t-test showed that ALP activity in the group which concentrations of simvastatin were 0.5 mumol/L (t = 2.35, P = 0.041), 1.0 mumol/L (t = 2.348, P = 0.041) had significant difference when compared with control group. CONCLUSION: Simvastatin lead to high expression of BMP-2 in bone marrow stromal cells, via the increased auto- or para-crine of BMP-2, and ALP activity increased. These may be parts of the mechanism on the anabolic osteogenetic effect of simvastatin.
【摘要】 目的 觀察百草枯中毒大鼠腎組織中血紅素氧合酶1(HO1)的表達,探討其病理生理機制。 方法 SD大鼠126只隨機分為空白對照組、中毒組和褪黑素組,各42只。中毒組、褪黑素組予以百草枯(25 mg/kg)腹腔注射染毒,對照組予以等量生理鹽水腹腔注射,15 min后褪黑素組予以褪黑素(10 mg/kg)腹腔注射,對照組、中毒組予以等量生理鹽水腹腔注射。于1、3、6、12 h,1、2、3、5 d時各組隨機取6只處死。蘇木精〖CD3/5〗伊紅（HE）染色觀察各組腎組織病理學變化,采用免疫組識化學和RTPCR觀察腎組織HO1蛋白和mRNA表達。 結果 ①與對照組相比,中毒組染毒后3 h即可見充血、水腫及空泡變性等病理變化,1 d達頂峰,病理損傷評分3.30±0.31(Plt;0.05),其后緩解趨勢不明顯；而褪黑素組病理變化明顯減輕且緩解趨勢明顯,1 d時病理損傷評分2.70±0.26,與中毒組相比差異有統計學意義(Plt;0.05)。②與對照組相比,中毒組染毒3 h在皮質部腎小管上皮細胞的細胞膜及細胞漿HO1呈陽性表達,免疫組識化學評分(IHS)3.33±1.75,HO1 mRNA表達增強,與對照組相比差異有統計學意義(Plt;0.05),1 d達頂峰,HIS為7.00±2.00,之后減弱,5 d仍有陽性表達,但與對照組相比差異無統計學意義(Pgt;0.05)；褪黑素組HO1表達較中毒組明顯增強,IHS評分6 h~3 d差異具有統計學意義(Plt;0.05),5 d不再有統計學意義(Pgt;005)。 結論 HO1在百草枯中毒大鼠腎組織中呈高表達,褪黑素能明顯改善百草枯中毒腎臟病理損傷,增強HO1表達可能是其作用途徑之一,而氧化損傷可能是百草枯中毒腎損傷病理生理機制之一。【Abstract】 Objective To investigate the expression of heme oxygenase1 (HO1) in paraquartinduced renal injury in rats and the protective effects of melatonin, and explore possible mechanism of paraquartinduced renal injury. Methods One hundred and twentysix adult healthy SpragneDawley rats were randomly divided into three groups and 42 in each group: control group (A), paraquart group (B), and melatonin group (C). The rats in group B and group C were treated with paraquart (25 mg/kg) intraperitoneally, the rats in group A were treated with the same dose of normal saline. In 15 minutes, the rats in group C were given melatonin intraperitoneally at a dose of 10 mg/(kg·d) and the rats in group A and B were treated intraperitoneally with the same dose of normal saline. Six rats in each group were randomly sacrificed at one, three, 12 hours and one, tou, three, five days respectively. Renal histopathological changes were observed under light microscope by HE staining. The protein and mRNA expressions of HO1 were evaluated by immunohistochemical staining and RTPCR respectively. Results ①In group B, there were obvious lesions in the renal tubule of cortical part, including edema, congestion and vacuolar degeneration. These pathologic changes gradually reached the peak on the first day and did not relieved till the end of this study, the pathologic injury score was 3.30±0.31, and there was a statistical significance between group B and group A (Plt;0.05). The aforementioned pathological lesion was more palliative in group C, the pathologic injury score was 2.70±0.26 at the first day; Compared with group B, there was a statistical significance. ②In group A, there was no or weak expression of HO1 and HO1 mRNA. At the third hour, the expression of HO1 in group B was observed in the membrane and cytoplasm of renal tubular epithelial cell of cortical part. Immunohistochemistry score (IHS) was 3.33±1.75 and the expression of HO1 mRNA increased, there was a statistical significance between group B and group A (Plt;0.05). It reached the peak on the first day, IHS was 7.00±2.00, but there was no statistical difference between group B and group A on the fifth day (Pgt;0.05); Compared with group B, the expression in group C was enhanced obviously, IHS were higher obviously on the six hour till to the third day (Plt;0.05), but there were no statistical differences on the fifth day (Pgt;0.05). Conclusion The expression of HO1 in the paraquartdamaged kidney increases and melatonin surely has an protective effect by increasing the expression of HO1, which suggests that oxidative injury might be the main mechanism of paraquartinduced renal injury.
Objective To explore effects of zinc on the contents of cycl in D2, cycl in-dependent kinase 4 (CDK4), and their DNA and total cellular protein in human umbil ical cord blood-drived mesenchymal stem cells (hUCBMSCs). Methods hUCBMSCs were isolated and cultured by density gradient centrifugation adherence method in vitro. At the serial subcultivation, the hUCBMSCs were randomly divided into 7 groups. In control group, hUCBMSCs were cultured with DMEM medium (containing 15%FBS). In treatment groups, hUCBMSCs were cultured with DMEM medium (containing 15%FBS plusZnSO4?7H2O). The final concentrations of zinc were 0.5, 1.5, 2.5, 3.5, 4.5, and 5.5 mg/L, respectively. The cellular surface antigens of CD29, CD34, CD44, and CD45 at the 3rd generation of hUCBMSCs were detected by flow cytometry. MTT assay was used to detect cell activity of the 3rd generation of hUCBMSCs. The optimum concentration of zinc was selected by the results of MTT as experimental group. The cell growth curves of experimental group and control group were drown by counting cell. The cell surface antigen, reproductive cycle, and DNA content were detected by flow cytometry motheds. The contents of cycl in D2 and CDK4 were detected by Western blot method. Results The positive expression rates of CD29 and CD44 were more than 70% in hUCBMSCs. The cell activity of 2.5 mg/L treatment group was superior to other treatment groups, as experimental group. At 7, 14, and 28 days, the contents of DNA, total cellular protein, cycl in D2, and CDK4 of hUCBMSCs were significantly higher in experimental group than those in control group (P lt; 0.01). The percentage of hUCBMSCs at S stage and prol iferation index in experimental group were also significantly higher than those in control group (P lt; 0.01). Conclusion Zinc (0.5-4.5 mg/L) has the promoting effect on the hUCBMSCs activity, and 2.5 mg/L is the optimal concentration. Zinc (2.5 mg/L) can accelerate the prol iferation and DNA reproduction of hUCBMSCs and increase the contents of cycl in D2 , CDK4, and cellular total protein.
ObjectiveTo understand the effect of nitric oxide (NO) on the formation of hyperdynamic circulatory syndrome (HCS) and the influence of level of NO on HCS. MethodsAfter establishment of stable HCS in partial portal vein ligated rats,the quantity of NO in blood of portal vein and the activity of nitric oxide synthase (NOS) in liver were determined by pre and post injection of inhabitor of NOS (NGmethylLarginine) and hemodynamics was supervised simultaneously.ResultsThe quantity of NO was paralleled with the activity of NOS and was elevated markedly by 24 hours after operation and reached the top by 48 hours after surgery. These sequential changes were coincided with the dilation of general vascularture. There was a close relation between this changes and the formation of HCS.The quantity of NO and the activity of NOS were decreased significantly to the level of the control group after injection of NGmethylLarginine (LNMMA). LNMMA inhabited the activity of NOS and blocked the production of NO. HCS ameliorated obviously. ConclusionNO plays an important role in initiating the dilation of general vascularture and plays a critical role in the formation of HCS. HCS will be ameliorated obviously or be blocked completely by eliminating the effect of NO and the portal pressure will decreased significantly or recover to normal range.
To investigate the function of nitric oxide (NO) and nitric oxide synthetase (NOS) inhibitor, N-nitro-L-arginine methyl ester (L-NAME), the skin avulsion model was made in the lower extremity of pig. The methods of measurement of size of the survived flap, weighing, immunocytochemistry and hybridization in situ were employed, so that the survival surface area of flaps, tissue wet/dry weight ratio, NO content in the serum, gene expression of NO and NOS content in the flap tissue were determined, respectively. The results showed that the early gene expression of NOS was increased as well as the NO content and tissue wet/dry weight ratio (P lt; 0.01). After L-NAME was applied introvenously, the NO content and tissue wet/dry weight ratio were decreased (P lt; 0.01), and the survival surface area of flaps was enlarged (P lt; 0.01). It could be concluded that the NO might play a role in the development of the pathological changes as early congestion, edema and secondary necrosis in the avulsed skin flaps. The early application of L-NAME could do some good to the avulsed skin flap and protect it from further necrosis owing to the presence of NO.
摘要:目的: 探討血清前白蛋白(prealbumin,PAB)、膽堿脂酶(cholinesterase,ChE)、總膽汁酸(total bile acid,TBA)在肝硬化中的檢測價值。 方法 :測定105例肝硬化患者和30例健康人的前白蛋白、膽堿脂酶、總膽汁酸活性及肝功能生化指標,并按ChildPugh分級進行比較。 結果 :肝硬化組前白蛋白含量、膽堿脂酶活性均較對照組顯著降低;按ChildPugh分級比較,肝硬化組前白蛋白含量在Child A級與對照組、B級與A級之間、在C級與B級之間差異有顯著性(Plt;001);膽堿脂酶活性在Child A級與對照組、B級與A級、C級與B級之間差異有顯著性(Plt;001)。總膽汁酸在Child B級與A級,C級與B級間有顯著差異性,在A級與對照組間差異無顯著性。 結論 :血清前白蛋白、膽堿脂酶在肝硬化早期評估中有很重要的價值,而總膽汁酸在肝硬化預后的判定中有重要價值。 Abstract: Objective: To evaluate the role of prealbumin (PAB), cholinesterase (ChE), and total bile acid (TBA) in evaluating liver reserve function in patients with liver cirrhosis. Methods : One hundred and five serum samples from patients with liver cirrhosis were detected in PAB, ChE, TBA and other biochemical markers. All patients were classified in accordance with ChildPugh scale. Results : For PAB, the differences among ChildPugh A, B, C and healthy group were statistically significant (t=1254, 1887, 2316) (Plt;001). For ChE, the differences among ChildPugh A, B, C and healthy group were statistically significant (t=1288, 0856, 1002) (Plt;001). For TBA, the differences among ChildPugh C group, B group and A group were statistically significant (t=0526, 1081)(Plt;001), the difference among ChildPugh A group and healthy group was not statistically significant (t=5615) (Pgt;005). Conclusion : PAB and ChE reflect liver reserve function earlier in patients with liver cirrhosis. The role of TBA is important in reflecting prognosis in patients with liver cirrhosis.
Objective To investigate the expression of urokinase-type plasminogen activator (uPA) mRNA in gastric cancer tissues and cancer-adjacent tissues and the relationship between its expression and biologic behavior of tumor. Methods Fourty-eight cases with gastric cancer were detected for the expression of uPA mRNA by fluorogenic probe quantitative reverse transcription polymerase chain reaction (RTPCR). Results The positive expression rate of uPA mRNA was 83.3%, 25.0%, 93.8% and 62.5% in gastric cancer tissues,cancer-adjacent tissues, gastric cancer tissues with lymph node metastasis and with non-lymph node metastasis respectively. Expression of uPA mRNA was positively related with the invasion depth of gastric cancer. Conclusion Expression of uPA mRNA is significantly increased in gastric cancer and it can be used as an indicator to judge the metastasis and prognosis of tumor.
【Abstract】Objective To study the antitumor activity of HSVtk/CD combinative gene toward human cholangiocarcinoma in vivo. Methods Nude mouse models with transplanted subcutaneous cholangiocarcinoma were constructed and divided into 4 groups randomly, each group had 8 mice. Adenovirus solution free from suicide gene was injected in subcutaneous tumors of each mouse of control group. Adenovirus solution containing cytosine deaminase (CD), thymidine kinase (tk) and HSVtk/CD fusional gene were injected into single suicide gene either HSVtk or CD was transinfected into the tumor cells by injecting viras into subcutaneous tumor of mice of CD gene,tk gene and fused CD and tk gene group respectively. 24 hours after the injection, 5fluorocytosine (5FC) and ganciclovir (GCV) were injected introabdominally in each mouse. Growth of the tumors were monitored.Results Tumor growth of the genetransfection groups was suppressed in different degrees. Compared with the control group, the suppressing rates of the genetransfection groups were 41.2%, 55.7% and 70.0% respectively (P＜0.05). Histological examination showed good tumor growth in the control group, and tumor necrosis in the other 3 groups, particularly obvious in the group transfected with pAd(HSVtk/CD).Conclusion Combinative gene system has a b antitumor effect on cholangiocarcinoma in vivo. But it’s not powerful enough to eliminate tumor thoroughly because of insufficient “Bystander effect”.
