Inhibition of 17-DMAG on VEGF expression in transplanted human gastric cancer in nude mice

AIM: To observe the anti-tumor effects of heat shock protein 90(HSP90) inhibitor 17-dimethylaminoethylamino-17 demethoxygeldanamycin (17-DMAG) on the tumor growth and angiogenesis in implanted gastric cancer nude mouse model. METHODS: Human gastric cancer cell HGC-27 was subcutaneous inoculation into the nude mice to develop a tumor model. Ten days later, 24 mice with implanted tumor were randomly divided into 3 groups: 17-DMAG group (receiving 17-DMAG at dose of 25 mg/kg), control group (treated with NS at dose of 10 mL/kg) and 5-fluorouracil(5-FU) group (treated with 5-FU at dose of 20 mg/kg). Four weeks after treatment, the tumor volume and weight, and the inhibitory rates of tumor growth were evaluated. In the meantime, the expression of CD31 was detected by immunohistochemical staining. The expression of vascular endothelial growth factor (VEGF) was determined by Western blotting. RESULTS: The size of xenografts in 17-DMAG treatment group was (288.10±23.32)mm³, and that in 5-FU treatment group was (366.37±26.42)mm³, both were significantly smaller than that in control group (957.66±117.51)mm³. The tumor weight in 17-DMAG treatment group was (0.41±0.02)g, significantly less than that in control group (1.12±0.08)g. The inhibitory rate of 17-DMAG was 63%. A significant decease of MVD in 17-DMAG group (21.72±1.24) was observed as compared to 5-FU group (36.70±1.51) and control group (37.78±1.68). The expression of VEGF in 17-DMAG group (15.39±4.37) was significantly lower than that in 5-FU group (26.11±6.26) and control group (36.45±7.45). CONCLUSION: HSP90 inhibitor 17-DMAG suppresses the expression of VEGF and the angiogenesis of the gastric cancer to inhibit the tumor growth.     

Polysaccharides of Dicliptera chinensis (L.) Nees. alleviates liver injury of GalN/LPS-induced fulminant hepatitis in rats

AIM: To investigate the liver protective effects of polysaccharides of Dicliptera chinensis (L.) Nees. on fulminant hepatitis caused by D-galactosamine (D-GalN)/lipopolysaccharide (LPS) in rats. METHODS: Male Wistar rats were divided into 4 groups (12 rats in each group), including normal control group, GalN/LPS-treated group, and two Dicliptera chinensis (L.) Nees. polysaccharides-treated groups (150 mg/kg, 300 mg/kg, ig, respectively). The Dicliptera chinensis (L.) Nees. polysaccharides and controlled normal saline (NS) were administered once 1 d for 6 d. One hour after the latest administration, all animals except for the animals in control group were administered with D-GalN (500 mg/kg, ip), then treated with LPS 1 h later. The mortality was observed at 6 h and 12 h after modeling. The serum samples were collected at 6 h and 12 h in the survival rats by retro-orbital sampling. All samples were measured for ALT, AST, TNF-α and IL-1β. The samples collected at 12 h were also detected for NO. At 12 h, all survival rats were sacrificed and liver section was prepared for histological evaluation. RESULTS: Pretreatment with polysaccharides of Dicliptera chinensis (L.) Nees. significantly improved the histopathological changes and attenuated GalN/LPS-induced severe hepatotoxicity as evidenced by decrease in the levels of ALT and AST. The polysaccharides of Dicliptera chinensis (L.) Nees. inhibited the elevated levels of TNF-α, IL-1β and NO. CONCLUSION: Using Dicliptera chinensis (L.) Nees. polysaccharides as anti-inflammatory reagent provides a definite protective way against fulminant hepatitis caused by GalN/LPS in rat.     

Effects of phillyrin on VEGF and endostatin expression in Lewis lung carcinoma

AIM: To investigate the effects of phillyrin on vascular endothelial growth factor (VEGF) and endostatin expression in lung tumor tissues isolated from Lewis lung carcinoma. METHODS: The expression of VEGF and endostatin in control individuals and the patients with lung cancer was determined by immunohistochemistry. In the animal experiment, 5 groups of animals were examined: control, tumor model, and tumor model with 3 different concentrations of phillyrin treatments. For preparation of transplanted tumor model, Lewis cells were subcutaneously injected into the right limb armpit of the nude mice. After that, phillyrin was administered via oral gavage once daily for 20 d at dose of 5 or 10 g/kg, or twice daily at 10 g/kg. Lung tumor tissues isolated from each group were observed by hematoxylin-eosin staining. VEGF and endostatin expression were examined by immunohistochemistry. RESULTS: VEGF expression was increased in lung tumor tissues as compared with normal and pericarcinous tissues, while endostatin expression was decreased. Phillyrin significantly inhibited the tumor size and tumor tissue density dose-dependently, which was accompanied with a decrease in VEGF expression and an increase in endostatin expression. CONCLUSION: Phillyrin inhibits the development of lung tumor through reducing VEGF expression and increasing endostatin expression.     

Changes of lung tissue and structure on bone marrow mesenchymal stem cells transplantation to treat the experimental pulmonary arterial hypertension in rats

AIM: To investigate the changes of lung tissue and structure on bone marrow mesenchymal stem cells (MMSCs) transplantation to treat the pulmonary arterial hypertension (PAH) in rats.METHODS: MMSCs cells were collected from bone marrow of SD rat’s femoral’s tibial bones, mix cultured with Hoechst 33342 fluorescence dye in vitro. Sixty healthy male Sprague-Dawley rats (weighing 180 g±5 g) were randomly divided into 3 groups: normal control group (group C), MMSCs transplanted group (group M), pulmonary model group (group H). The rats of the two latter groups were given a single subcutaneous monocrotaline (50 mg/kg) to induce the model of PAH. The rats of normal control group were injected respectively a single subcutaneous isotonic Na chloride (6 mL/kg). The three groups were fed in the same condition. After 3 weeks, 5×109 cells/L MMSCs with l mL phosphate-buffered saline were infused into the rats respectively in group M by sublingual vein and 1 mL L-DMEM was administered in group H. Nothing was given to group C. The viability, right ventricle systolic pressure (RVSP), right ventricle hypertrophy index, the microstructure and ultrastructural changes of small pulmonary vascular were observed after 28 d.RESULTS: The viability of administrated MMSCs 28 d after PAH was 100% in transplanted group, and it nearly completely prevented the increase in right ventricular systolic pressure (RVSP) compared to PAH alone ［(32.20±2.32)mmHg vs (48.30±1.56)mmHg, respectively, P<0.05］, right ventricle hypertrophy index also degraded (38.80%±3.24% vs 45.10%±3.43%, respectively, P<0.05). The pulmonary arteries remodeling and ultrastructural changes of lung, such as blood gas barrier, chondriosome, osmiophilic lamellar body and so on, were improved significantly in MMSCs transplanted group. The changes were similar to the results of normal control group, but they were better than that in pulmonary arterial hypertension model group, which had conspicuous significance of statistics. MMSCs labeled with Hoechst 33342 were observed that they permanent planted and differentiated into plentiful collateral circulations in lung observed by fluorescence microscope. The pulmonary arteries remodeling were attenuated effectively in MMSCs transplanted group. Pulmonary fibrosis and tumorous were not observed in rats of MMSCs transplanted group.CONCLUSION: These results indicate that MMSCs reduce and even reverse the progression of pulmonary arterial hypertension (PAH) induced by crotaline. It restores the structure and function of microvasculature with marked improvement of tissue and structure changes of the lung.     

The inhibitory action of rhTRAIL on mouse breast carcinoma

AIM:To explore the inhibitory action of recombinant human tumor necrosis factor-related apoptosis-inducing ligand(rhTRAIL) on mouse breast cancer.METHODS:Each mouse was inoculated 0.2 mL (1×106) D2F2 cells subcutaneously in the right lower limb and they were divided into five groups randomly.The control group was infused PBS 0.2 mL, while the low-dose, medium, high groups received purified rhTRAIL 2.5 mg/kg, 5.0 mg/kg, 10.0 mg/kg, respectively, the positive group was administered cyclophosphamide 30.0 mg/kg.Every group was operated by peritoneal injection once a day for fifteen days.The mice were weighed every day.The growth state was viewed and the size of the tumor was measured every 3 d to calculate the tumor volume and tumor suppression rate.All mice were killed after 15 d.The pathologic changes of the tumor were observed under light-microscopy and electronic microscopy.The cell cycle and apoptosis index of D2F2 cells were analyzed by flow cytometry.RESULTS: The body weight and tumor volume in low-dose, medium, high groups were lower than those in control group and the restriction effect was more significant than that in the control group (P<0.01).The body weight and tumor volume in low-dose, medium, high groups decreased with the increase in rhTRAIL concentration.The difference was significant (P<0.01).Both apoptosis and necrosis of tumor cells were observed in low-dose, medium, high groups.The area of cell apoptosis was large and most area didnt have tumor cells but only apoptotic bodise.The results of flow cytometry showed that rhTRAIL induced the apoptosis of D2F2 cells, and the apoptosis rates were 7.56 %, 21.37 %, 27.16 % respectively when rhTRAIL doses were 0.25 mg/L, 0.5 mg/L, 1.0 mg/L.Significant differences among three groups were observed (P<0.05).CONCLUSION:rhTRAIL induces apoptosis in mouse breast cancer cells and even the necrosis of tumor cells.rhTRAIL has significant effect on inhibiting the growth of D2F2 cells.     

Effect of neural stem cells transfected with VEGF on expression of NSE in rat brain injury irradiated by radioaction

AIM: To explore the effect of neural stem cells (NSC) transfected with vascular endothelial growth factor (VEGF) on brain injury irradiated by radioaction. METHODS: Mature Sprague-Dawley (SD) rats were randomly divided into 4 groups: control group, radiation brain injury group, NSC therapeutic group, NSC transfected with VEGF (NSC/VEGF) therapeutic group. NSC transfected with VEGF was transplanted into the rat brain of therapeutic group after preparation of radiation encephalopathy model for 1 week. The brain tissue was taken at week 1, 2, 3, 4 and 6 after transplantation. Immunohistochemical method was used to assay the count of neuro-specific enolase (NSE) stained cells in brain tissue. Western blotting was used to detect the expression of NSE protein. RESULTS: Compared to control group, the count of NSE stained cells in brain of radiation brain injury group was decreased significantly (P<0.05). Compared to that in radiation brain injury group, the count of NSE stained cells in brains was enhanced in NSC therapeutic group and NSC/VEGF therapeutic group since 3 weeks after transplantation (P<0.05). Compared to that in NSC therapeutic group, the count of NSE stained cells in NSC/VEGF therapeutic group was more enhanced since 6 weeks after transplantation (P<0.05). Compared to that in radiation brain injury group, the expression of NSE protein was enhanced in NSC group and NSC/VEGF group since 3 weeks after transplantation (P<0.05). Compared to that in NSC group, the expression of NSE protein in NSC/VEGF group was more enhanced since 6 weeks after transplantation (P<0.05). CONCLUSION: Transplantation of NSC transfected with VEGF may increase the content of NSE in rat brain of radiation encephalopathy model.     

VPA alleviates bleomycin-induced pulmonary fibrosis in rats

AIM: To investigate the effect of sodium valproate (VPA) on bleomycin-induced pulmonary fibrosis (PF) and its mechanism. METHODS: SD rats (n=42) were randomly divided into model group and treatment group. Bleomycin at dose of 5 mg/kg was intratracheally injected to establish a rat PF model. The rats in treatment group were given normal saline (NS, 0.5 mL/d), VPA (300 mg·kg^-1·d^-1) or dexamethasone (DEX, 0.6 mg·kg^-1·d^-1) via intraperitoneal injection from the 14th day to the 28th day after modeling. The rats in model group were sacrificed on 0 d, 14 day and 28 d after modeling . The rats in treatment group were killed at 14th day after treatment. The effects of VPA on PF were evaluated by HE and Masson staining. The hydroxyproline (HYP) content in the rat lung tissues was detected, and the expression of α-smooth muscle actin (α-SMA) and E-cadherin was determined by Western blotting. RESULTS: HE staining showed that the alveolar structure and interstitial morphology in VPA group were better than those in NS group and DEX group. The level of collagen in VPA group was significantly lower than that in DEX group and NS group by determining the HYP content and Masson staining. VPA reduced the expression of α-SMA, a mesenchymal marker protein of PF, while increased the expression of epithelial marker protein E-cadherin. CONCLUSION: VPA reduces collagen content and distribution, and up-regulates the expression of the epithelial marker protein E-cadherin, while down-regulates mesenchymal marker protein α-SMA, thereby preventing the rat lung tissues from bleomycin-induced PF.     

Protective effect of dexmedetomidine-ulinastatin combination on lipopolysaccharide-induced acute lung injury in rats

AIM：To investigate the effects of dexmedetomidine-ulinastatin combination on acute lung injury induced by lipopolysaccharide (LPS) in rats. METHODS：Male Wistar rats were randomly divided into 5 groups: saline control group (NS group) was given saline (5 mL/kg, iv) alone; LPS group (L group) was given LPS (10 mg/kg, over 10 min); dexmedetomidine+LPS group (L+D group) was treated with the additional administration of dexmedetomidine (1 μg·kg -1·h -1) immediately after LPS injection; ulinastatin+LPS group (L+U group) was treated with the addi-tional administration of ulinastatin (50 000 U/kg, ip) immediately after LPS injection; dexmedetomidine+ulinastatin+LPS group (L+D+U group) received dexmedetomidine (1 μg·kg -1·h -1) and ulinastatin (50 000 U/kg) immediately after LPS injection. The animals were sacrificed at 6 h after LPS or NS administration. Partial pressure of arterial oxygen (PaO 2), pH and base excess (BE) were measured, and the lungs were removed for evaluation of histological characteristics and determining the concentrations of TNF-α, IL-1β, macrophage inflammatory protein 2 (MIP-2), malondialdehyde (MDA), nitric oxide (NO), prostaglandin E 2 (PGE 2) and myeloperoxidase (MPO) in lung tissues, lung wet/dry weight ratio (W/D), and albumin in brochoalveolar lavage fluid (BLAF). The pulmonary expression of nuclear factor kappa B (NF-κB) p65 was evaluated by Western blotting. RESULTS：Compared with NS group, PaO 2, pH and BE was lower in L group, which was increased by treatment with dexmedetomidine-ulinastatin combination but not by dexmedetomidine or ulinastatin alone. Compared with NS group, LPS induced marked lung histological injury, which was less pronounced in the animals treated with dexmedetomidine-ulinastatin combination but not dexmedetomidine or ulinastatin alone. The levels of IL-1β, IL-6, MIP-2, MDA, NO and PGE 2 in the lung tissues increased in L group compared with NS group, which were reduced by dexmedetomidine-ulinastatin combination but not by dexmedetomidine or ulinastatin alone. The MPO activity, MDA level and W/D increased in the lung tissues in L group compared with NS group, which was reduced by dexmedetomidine-ulinastatin combination but not by dexmedetomidine or ulinastatin alone. Compared with NS group, the albumin concentration in the BLAF increased, which was reduced by dexmedetomidine-ulinastatin combination but not by dexmedetomidine or ulinastatin alone. Compared with NS group, the expression of NF-κB p65 increased in L group, which was reduced by dexmedetomidine-ulinastatin combination but not by dexmedetomidine or ulinastatin alone.CONCLUSION：Dexmedetomidine-ulinastatin combination has a protective effect on LPS-induced acute lung injury in the rats.     

Evaluation of the inflammatory response in two-hit acute lung injury model of rats using ［8F］ FDG microPET

AIM: To investigate whether two-hit acute lung injury (ALI) model is better than one-hit, and to evaluate the inflammatory response in the lungs during these models by using ［8F］FDG microPET. METHODS: Thirty three, adult, male Sprague-Dawley rats weighing 180-210 g were used and divided into 4 groups. Rats in LPS group (n=10) and LPS-HCl group (n=10) were challenged with intraperitoneal administration of LPS at the dose of 5 mg/kg, while rats in NS group (n=3) and HCl group (n=10) received normal saline solution intraperitoneally at the dose of 1 mL/kg, after 16 h, all animals were anesthetized with an intraperitoneal injection of sodium pentobarbital (40 mg/kg) and placed in a 60°inclined position, the femoral artery was cannulated and connected to a pressure transducer to record the arterial pressure on a polygraph recorder, the trachea was surgically exposed. Rats in HCl group and LPS-HCl group received direct intratracheal injection of HCl (pH=1.2) at the dose of 0.5 mL/kg while rats in NS group and LPS group received the same volume of normal saline solution. Blood gas samples (each 0.3 mL) were obtained at 30, 90 and 240 min after the instillation and replaced by the same volume of saline solution, the samples were analyzed using a blood gas analyzer. 240 min after HCl or NS administration, the rats underwent a microPET scanning, then, all the rats were sacrificed and the lungs were obtained for histological analysis. RESULTS: Blood gas analysis showed that rats in LPS-HCl group had higher PaO2 and lower PaCO2 than the other groups. MAP decreased markedly in LPS-HCl group, while MAP in other groups remained stable. The results of microPET showed that the ratio of ROI between the right lung and the muscle tissue of the right arm in LPS-HCl group was 9.00±1.41, and was significantly higher than that in LPS group (4.01±0.60) and HCl group (3.33±0.55). Histological examination showed that the mean lung injury score in LPS-HCl group was 12.70±0.95, while that was 8.40±1.26 in HCl group and 7.00±0.82 in LPS group, and there were significant differences (both P<0.01). CONCLUSION: LPS pretreatment significantly magnifies and prolongs the inflammatory response to the subsequent acid instillation in both lungs. When compared with “one-hit”, “two-hit” is easier to induce the ALI, and ［8F］FDG microPET is a useful tool to evaluate the inflammatory reaction during ALI.     

Effect of hydroxylamine on pulmonary arterial hypertension induced by chronic hypoxic hypercapnia in rats

AIM: To explore the effects of hydroxylamine on the pulmonary arterial pressure in chronic hypoxic hypercapnic rats. METHODS: Twenty-four male Sprague-Dawley rats were randomly divided into 3 groups (8 rats in each group): the normal control group (NC), hypoxic hypercapnia+normal saline group (NS), hypoxic hypercapnia+hydroxylamine group (HA). The animals in NS and HA groups were kept in the O₂ (9%-11%) and CO₂ (5%-6%) cabin, 8 h a day and 6 days a week for 4 weeks. Before entering the cabin, the rats in HA group were administered with 1 mL hydroxylamine (12.5 mg/kg) by intraperitoneal injection, while the rats in NS group were given intraperitoneal injection of 1 mL saline solution. The mean pulmonary arterial pressure (mPAP) was measured by external jugular vein cannulation. The heart was removed, and the right ventricle (RV) and the left ventricle plus the septum (LV+S) were dissected. The ratio of the wet weight of the RV to that of the LV+S was calculated. The changes of the pulmonary vascular construction were observed under optical microscope. The concentration of H₂S in the plasma was measured with a spectrometer. The expression of cystathionine-γ-lyase (CSE) in the pulmonary arterioles and bronchi was measured by immunohistochemistry and RT-PCR. RESULTS: The values of mPAP, RV/(LV+S),vessel wall area/total area (WA/TA) and media thickness of pulmonary arterioles (PAMT) in NS group and HA group were significantly higher than those in NC group (P<0.05). The level of H₂S in the plasma, the content of CSE protein and the expression of CSE mRNA in NC group were significantly lower than those in NS group (P<0.05). The values of mPAP, RV/(LV+S), WA/TA and PAMT in HA group were significantly lower than those in NS group (P<0.05). The level of H₂S in the plasma, the content of CSE protein and the expression of CSE mRNA in HA group were significantly higher than those in NS group (P<0.05). CONCLUSION: Hydroxylamine may decrease the pulmonary arterial hypertension induced by chronic hypoxic hypercapnia in rats by increasing the level of H₂S in the plasma, the content of CSE protein and the mRNA expression of CSE, thus improving the pulmonary vascular structural remodeling.     

NK cells from donor liver inhibit NF-κB activity and IL-15 expression after liver transplantation in rats

AIM:To investigate the mechanism that donor liver natural killer (NK) cells alleviate acute rejection after liver transplantation by observing the secretion level of interleukin 15 (IL-15) in peripheral blood, the protein expression of IL-15 in transplanted liver tissues and the activity of NF-κB in spleen tissues in rat acute liver graft rejection model. METHODS:An acute rejection model of liver transplantation in rats was established by the modified two-cuff method, in which Lewis rats were used as donors and BN rats as recipients. The donor leukocytes were depleted by whole body irradiation of ［60Co］ source and the donor liver immunity was reconstituted by transfusion of liver NK cells from the same type of donor (donor type liver NK cells, dtlNKs) via portal vein immediately after grafting the irradiated liver. The rats were divided into the following groups: group A, acute rejection group; group B, BN rats receiving the liver of Lewis rats with ［60Co］ irradiation; group C, BN rats receiving the liver of ［60Co］-irradiated Lewis rats and treated with dtlNKs via the portal vein. The recipients were sacrificed at 1 d, 3 d and 7 d after transplantation. IL-15 level in peripheral blood was detected by ELISA. The expression of IL-15 in the liver grafts was determined by Western blotting. NF-κB activity in the spleen tissues of recipient rats was identified by electrophoretic mobility shift assay. The survival quality and living time in crude survival subgroup were observed. RESULTS:Acute rejection in group B was severer than that in group A and group C. The rats in group B showed significantly shorter average survival time compared with group A and group C. At 3 d and 7 d after transplantation, the IL-15 content in peripheral blood was significantly higher in group B than that in group A and group C. The expression of IL-15 in transplanted liver tissues was significantly higher in group B than that in group A and group C. The activity of NF-κB in the spleen tissues was higher in group B. CONCLUSION: IL-15 might be a significant indicator for monitoring acute rejection after liver transplantation. The donor liver NK cells modulate the immunity of liver transplantation by inhibiting the expression of IL-15 via the suppression of NF-κB activity.     

Effect of microcapsulated catechin on expression of vascular endothelial growth factor in rats with adriamycin induced-nephrotic syndrome

AIM: To study the effect of microcapsulated catechin on vascular endothelial grower factor (VEGF) expression in rats with adriamycin induced-nephrotic syndrome.METHODS: 120 female SD rats were randomly distributed in control group,nephrotic group,dexamethason group,vitamin E group,catechin group and microcapsule group.Rat with nephrotic syndrome were induced by injection of adriblastine (5 mg/kg BW).VEGF concentrations in serum and urine were detected by ELISA assay.VEGF expression in kidney was measured by immunohistochemistry assay.RESULTS: At the end of 4th week and 6th week,VEGF concentration in other groups in kidney,serum and urine were higher than that in control (all P<0.01),and were lower than that in nephritic group (exclude Vit E group,all P<0.01).Serum and urine VEGF concentrations in microcapsule group were lower than those in catechin group (P<0.01,P<0.05,respectively).VEGF expression in microcapsule group in kidney was lower than that in catechin group,but it was not significant different.Urinary protein excretion at 24 h in microcapsule group was lower than that in catechin group (P<0.05),there was positive correlation among urine protein,serum VEGF level,urine VEGF concentration and renal VEGF expression at 24 h (all P<0.01).CONCLUSION: Catechin decreases urinary protein excretion in the rats with nephrotic syndrome through reducing the expression and secretion of VEGF.Microcapusulated catechin is benefit in decreasing the expression and secretion of VEGF.     

Survival and neuronal differentiation of transplanted stem cells derived from embryonic hippocampus in adult rats with stroke lesions

AIM: To study whether exogenous neural stem cells (NSCs) derived from embryonic hippocampus differentiates into the neurons after transplanted into the infarct periphery of the brain in a stroke model and to further investigate the behavioral improvement in the rats.METHODS: The NSCs were prepared after isolated from the embryonic hippocampus of green fluorescent protein (GFP)-transgenic rats and cultured. The NSCs were identified using nestin and doublecortin(DCX) as markers. The cortical infarction in rats was induced using photochemical method, named photothrombotic cortical injury (PCI). Twenty adult rats were randomly divided into NSC transplantation group (NSC group) and control group. The cultured NSCs were transplanted into the infarct periphery of the rats in NSC group, and nothing in control group at first day was applied after PCI. The locomotor behavior of animals was checked using the rotarod test at 1st, 7th, 14th and 21st d after PCI. The survival and differentiation of transplanted NSCs were evaluated by immunocytochemical method at 12th week after PCI.RESULTS: The cells derived from the embryonic hippocampus significantly expressed the markers of NSCs. The grafted cells survived at least 12 weeks in the infarct periphery of adult rats and differentiated into mature glial cells and neurons. The density of NeuN+/GFP+ and volume of grafts at 12th week were less than those at 3rd week after transplantation (P<0.05). The time standing on the rod was longer in NSC group than that in control group at 7th, 14th and 21st d after PCI (P<0.01).CONCLUSION: The NSCs derived from embryonic hippocampus survive in the infarct periphery of adult rats up to 12 weeks and differentiate into mature neurons, which might be associated with the improvement of locomotor behavior of stroke animals. The neuronal replacement of neurons differentiated from NSCs may be the underlying mechanism.     

龙牙蕉组织培养技术

龙牙蕉吸芽茎尖经MS＋6-BA5．0mg／L＋AD20mg／L培养基诱导出不定芽后,在MS＋BA4mg／L＋NAA0．1mg／L培养基上继代培养对不定芽的分化较好,平均分化倍数为2．6倍以上,无根小苗在1／2MS＋NAA0．5mg／L＋AC1g／L培养基上进行生根培养,1周后长出新根。组培小苗移栽假植成活率94％,大田栽培表现正常。     

Effect of ischemia or hypoxia on vascular endothelial growth factor production in rat myocardium and its significance

AIM:To determine the relationship between ischemia, hypoxia and the production of vascular endothelial growth factor in rat myocardium and its basic mechanism. METHODS:(1) 28 Wistar rats were randomly divided into 4 groups: group A, normal control;group B, 1 day's acute myocardial infarction;group C, 3 day's acute myocardial infarction;group D, 7 day's acute myocardial infarction. (2) Rat cardiac myocytes cultured were primarily divided into some groups, hypoxia incubated 24 hours; PMA groups, hypoxia incubated 24 hours with PKC activator (PMA), A 0 ng/mL; B 10 ng/mL; C 100 ng/mL; D 1 000 ng/mL; Chelerythrine groups, hypoxia incubated 24 hours with PKC inhibitor (chelerythrine), A 0 nmol/L; B 10 nmol/L. (3) By computer scanned and quantitated, vascular endothelial growth factor (VEGF) protein was detected with immunohistochemical technique. RESULTS:The longer time of ischemia and hypoxia was, the higher the VEGF production.The relat ionship was found between the time of ischemia or hypoxia and the production of VEGF.The product ion of VEGF protein was further promoted by PMA with different concentrat ion, decreased by chelerythrine.CONCLUSION: Ischemia or hypoxia strongly stimulated the production of VEGF in myocardium, which played an important role in autoprotecting of ischemic or hypoxic myocardium. Hypoxia-induced PKC activation is one kind of basic mechanisms in this course.     

Effect of sorafenib on liver regeneration after partial hepatoectomy in liver cirrhotic rats

AIM: To study the effect of sorafenib on the liver regeneration after partial hepatectomy (PH) in cirrhotic rats. METHODS: Thirty Wistar rats with liver cirrhosis induced successfully with diethylnitrosamine (DEN) underwent 30% PH and then were randomly divided into 2 groups (n=15). The rats in experimental group were fed with sorafenib at dose of 30 mg·kg^-1·d^-1 from the 1st day to the 10th day after PH, while those in control group were fed with vehicle by gavage. The blood and liver tissues of the rats were collected after PH and at the end of the experiment. Liver regeneration rate (LRR) and proliferating cell nuclear antigen (PCNA) expression were assessed for determining the hepatocyte proliferation. The content of alanine transaminase (ALT), albumin (ALB), total bilirubin (TBIL), direct bilirubin (DBIL), angiogenesis related factors including vascular endothelial growth factor (VEGF), vascular endothelial growth factor receptor 2 (VEGFR-2), platelet-derived growth factor receptor β (PDGFR-β) and micro-vessel density (MVD) were measured in both groups. RESULTS: LRRs on day 10 after PH were 45.43%±3.36% and 44.21%±2.77% in experimental group and control group, respectively (P>0.05), and the expression of PCNA in hepatic tissues of the rats was not found by the method of immunohistochemistry in both groups. Liver function index had no significant difference between the 2 groups (P>0.05). However, other than VEGF, sorafenib resulted in inhibition of VEGFR-2 and PDGFR-β expression and reduction of MVD in experiment group, and significant difference between the 2 groups was observed (P<0.01). CONCLUSION: Sorafenib does not influence live regeneration after PH in liver cirrhotic rats.     

姬松茸发酵液对小鼠抗运动疲劳及肝组织抗氧化能力的影响

研究姬松茸(Agaricus blazei)发酵液灌胃小鼠,对小鼠抗运动疲劳及肝组织抗氧化能力的影响.试验结果表明,与对照组(10 mg/kg·d ddH2O)相比,姬松茸发酵液中剂量组(20mg/kg·d)、高剂量组(30 mg/kg·d)小鼠的负重游泳、爬杆、耐缺氧时间均显著延长,血清尿素氮和血乳酸含量显著降低,肝糖原储存量显著升高,肝组织中SOD、GSH-Px活性显著提高,MDA值显著下降.说明灌胃姬松茸发酵液,能提高小鼠抗运动疲劳功能和加强肝组织抗氧化能力.     

Effects of spermine on myocardial ischemia/ reperfusion injury in rats

AIM: To observe the effect of exogenous spermine (low concentration) on myocardial ischemia/reperfusion injury in rats.METHODS: 40 Wistar rats were randomly divided into 4 groups: sham- operation group (Sham), ischemic reperfusion group (I/R), spermine group (Sp) and natural saline group (NS). The model of ischemic/reperfusion injury was established by ligating rat coronary artery. In Sp group, spermine (0.5 mmol/L, 2 mL/kg) was injected slowly into rat vein. During the process, we recorded the electrocardiogram and the LV functional parameters, assayed the levels of SOD, LDH, NO and MDA in serum, and examined the ultrastructure of the myocardium. RESULTS: In I/R group, the incidence of arrhythmia was 90%, myocardial ultrastructure was injured seriously, values of LVSP and ±dp/dtmax decreased, levels of LDH, NO and MDA increased while SOD activity decreased (P<0.05 or P<0.01, compared with Sham group). Compared with I/R and NS group, all those indexes in Sp group changed significantly (P<0.05 or P<0.01). CONCLUSION: Exogenous spermine alleviates myocardial ischemia/ reperfusion injury in rats. The mechanism may be related to its antioxidant effect and relieving the injury caused by oxygen free radical.     

Effect of rhSOD on pulmonary nuclear factor-kappa B and MIP-1α expression in meconium-induced acute lung injury

AIM: To evaluate the role and mechanisms of recombinant human superoxide dismutase (rhSOD) in meconium-induced acute lung injury (ALI) by evaluating pulmonary MIP-1α and NF-κB expression. METHODS: 24 health male Sprage-Dawley rats were randomized to 3 groups (8, each group), followed by intratracheal (IT) administration with (1) saline at 1 mL/kg (control group); (2) 20% human newborn meconium suspension at 1 mL/kg, followed by saline at 1 mL/kg (Mec/saline group); (3) 20% human newborn meconium suspension at 1mL/kg, followed by rhSOD at 20 mg/kg (Mec/rhSOD group). The animal was killed 24 h after treatment. The measurements included the bronchoalveolar lavage (BAL) cell count, RT-PCR analysis of pulmonary MIP-1α mRNA expression, Western blotting analysis of pulmonary NF-κB expression. RESULTS: Meconium-induced ALI was characterized by increased BAL cell count, increased expressions of pulmonary MIP-1α mRNA and NF-κB protein ［(4.68±1.40)×109 cells/L vs (0.53±0.19)×109 cells/L, 3.60±0.75 vs 1.56±0.33, 0.72±0.31 vs 0.23±0.12, respectively in control rats, all P<0.01］. IT administration of rhSOD early in the ALI rat significantly decreased meconium-induced BAL cell count ［(3.13±0.77)×109 cells/L vs (4.68±1.40)×109 cells/L in Mec/saline rats, P<0.01］, inhibited the expression of pulmonary MIP-1α mRNA (2.20±0.39 vs 3.60±0.75, in Mec/saline rats, P<0.01) and NF-κB protein (0.44±0.21 vs 0.72±0.31 in Mec/saline rats, P<0.05). CONCLUSION: The early IT administration of rhSOD in ALI rat following meconium aspiration protects lung from inflammatory injury through inhibiting meconium-induced pulmonary MIP-1α mRNA and NF-κB protein expression.     

Effects of WJMSC transplantation on cesarean scar defect in a rat model

AIM: To evaluate the effects of transplantation of Wharton jelly-derived mesenchymal stem cells (WJMSCs) on SD rats with previous cesarean scar defect (PCSD). METHODS: SD rats (n=50) were randomly divided into 3 groups:10 in normal operation group (N-O group), 20 in model control group (M-C group) and 20 in model treatment group (M-T group). On the 19th day of pregnancy, thread was used for continuous suture after the caesarean section (C-S) in N-O group. After C-S, LPS (1 g/L, 1 mL/kg) was injected in uterine muscle walls and electrocoagulation (power=10 W) was used in bilateral uterine incision with interrupted suture in both M-C and M-T groups. The rats in M-T group was further divided into A and B subgroups, with 10 rats each. The rats in group A were treated by intravenous injection and vaginal perfusion, both with 0.5 mL of 1×10⁹/L WJMSCs, while the rats in group B were treated by vaginal perfusion with 1 mL of WJMSCs, once a week, totally 3 times. The rats in M-C group were also divided into A and B subgroups (10 rats per group) and treated by the same way as M-T group, only except that saline was used instead of WJMSCs. At the 1st and 4th weeks after the treatment was finished, 6 and 4 rats were executed in every group, respectively. The uterine specimens with Masson and immunohistochemical staining were observed for determining the expression of actin and cytokeratin to reveal the repair of endometrial epithelium. RESULTS: The thickness of uterine muscle and volume density of actin in M-C group were significantly lower than those in N-O group, but the muscle collagen volume density and defect rate of endometrium were higher than those in N-O group. At the 1st and 4th weeks after treatment with WJMSCs, the muscular thickness and muscular actin concentration in M-T group (A and B) were significantly higher than those in M-C group, while the number of muscular collagen fibers were significantly lower than that in M-C group. The endometrial insufficiency in M-C group was higher than that in the other groups (P<0.05). CONCLUSION: WJMSCs promote the repair of the structure and function of myometrium in PCSD rats. No obvious difference between the effects of intravenous injection combined with vaginal perfusion, and vaginal perfusion alone on the repairing is observed.