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】目的探討重癥急性胰腺炎(SAP)時胰腺組織的誘導型一氧化氮合成酶(iNOS)、內皮素（ET1） mRNA表達狀態, 以及與血漿中NO、ET1濃度和腸道損傷的關系及丹參治療的影響。方法Wistar大鼠45只隨機分為3組:SAP模型組(A組),SAP丹參治療組(B組),假手術 組(C組),進行不同治療和觀察分析。結果A組血中淀粉酶(AML)、ET1、NO、內毒素(LPS)含量、125 I白蛋白累積指數及腹水量均顯著高于C組(Plt;0.01);與A組比較,B組胰腺ET1和iNOS mRNA表達較弱,血中AML、ET1、NO、LPS及腹水量顯著下降(Plt;0.01),125 I白蛋白累積指數較A組也有下降,但無差異(Pgt;0.05)。結論SAP時存在腸道損傷,胰腺組織ET1、iNOS mRNA的過度表達,使血中ET1、NO濃度升高,造成腸道屏障功能受損,腸通透性增加,引起內毒素血癥。丹參注射液通過減輕SAP時胰腺的病理損害程度,下調胰腺ET1和iNOS mRNA的表達,使血中ET1、NO濃度下降,對SAP及其腸道損傷有一定治療作用。
Objective To explore the effects of heat-inactivated Lactobacillus gasseri TMC0356 on liver lipid metabolism in rats with metabolic syndrome (MS) and its possible mechanism. Methods Sixty male Sprague-Dawley rats were selected. Rats were randomly divided into 5 groups, including control group, MS model group and three TMC0356 test groups (low-, medium- and high-dose groups). The rats in each group were fed with different diets for 7 days, and the liver was dissected and removed after 15 weeks. The mRNA and protein expression levels of peroxisome hyperbioactive receptor-α (PPAR-α), sterol regulatory element binding protein-1c (REBP-1c), fatty acid synthase (FAS) and carnitine lipoacyltransferase-1 (CPT-1) genes in liver were detected. Results There was no significant difference in the mRNA expression of PPAR-α, SREBP-1c or CPT-1 among the five groups (P>0.05). The mRNA expression of FAS in low-dose TMC0356 test group was lower than that in MS model group (P=0.011), medium-dose TMC0356 test group (P=0.042) and high-dose TMC0356 test group (P=0.009). There was no significant difference in the expression of FAS mRNA between other groups (P>0.05). There was no significant difference in the protein expression of PPAR-α, SREBP-1c or FAS among the five groups (P>0.05). The protein expression of CPT-1 in low-dose TMC0356 test group was higher than that in control group (P=0.033) and high-dose TMC0356 test group (P=0.043). There was no significant difference in the protein expression of CPT-1 between the other groups (P>0.05). Conclusion Heat-inactivated Lactobacillus gasseri TMC0356 may improve the symptoms of metabolic disorder in rats by suppressing appetite, improving insulin resistance, and downregulating the expression of key fat metabolism genes such as FAS and SREBP-1c.
Objective To determine the genotype distribution of methylenetetrahydrofolate reductase (MTHFR) C677T, A1298C and methionine synthase reductase (MTRR) A66G involved in the folic acid biosynthetic pathway among Chinese Han women in Sichuan, so as to provide pregnant women with guidance of supplementing folic acid. Methods By means of Taqman-MGB, 2382 samples from Deyang region in Sichuan province were tested for detecting the genotype distributions and allele frequencies of MTHFR C677T, MTHFR A1298C and MTRR A66G polymorphisms, and then the results were compared with published data in Shandong, Henan and Hainan provinces. Results The allele frequencies of MTHFR C677T were 63.45 and 36.55, those of MTHFR A1298C were 78.20 and 21.80, and those of MTRR A66G were 72.81 and 27.19. There were significant differences in allele distribution of MTHFR C677T and A1298C between Sichuan Han women and other region population. Conclusion This study suggests that the polymorphism of MTHFR C677T and A1298C exhibits region heterogeneity. The polymorphisms of MTHFR may play a role in neural tube defects (NTDs) risk, so periconceptional folic acid supplementation and healthcare following gene polymorphism testing may be a powerful measure to decrease congenital malformations of the central nervous system.
ObjectiveTo investigate the effects of 3-(5'-hydroxymethyl-2'-furyl)-1-benzylindazole (YC-1), a hypoxia-inducible factor-1α (HIF-1α) inhibitor, on hypoxia induced rat pulmonary arterial adventitial fibroblasts (AFs) proliferation and collagen synthesis, and explore the molecular mechanism.MethodsUnder hypoxic condition, rat AFs were cultured in DMEM medium supplemented with 10% fetal bovine serum in vitro. The cells were divided into five groups, ie. a normoxia group, a hypoxia group and three hypoxia+YC-1 groups (treated with YC-1 at concentration of 0.01, 0.05 and 0.1 mmol/L, respectively). The cells proliferation was determined by MTT method. Collagen synthesis of AFs was measured by 3H-proline incorporation assay. The expression of HIF-1α in AFs in different conditions was measured by Western blot, and the mRNA expression of transforming growth factor-β1 (TGF-β1) was measured by reverse-transcription polymerase chain reaction.ResultsThe proliferation rate and the incorporation data of 3H-proline in the hypoxia group were significantly increased as compared with those in the control group (both P<0.01). YC-1 significantly reduced the proliferation rate and incorporation data of3H-proline induced by hypoxia in a dose-dependent manner. YC-1 could also down-regulate the expressions of HIF-1α and TGF-β1 mRNA significantly (both P<0.01). Compared with the hypoxia group, the expressions of HIF-1α and TGF-β1 mRNA decreased respectively by 65% and 61% in the hypoxia+YC-1 (0.1 mmol/L) group (bothP<0.01).ConclusionsYC-1 can inhibit hypoxia-induced AFs proliferation and collagen synthesis in a dose-dependent manner. The mechanism may relate to YC-1’s inhibitory effect on expressions of HIF-1α and TGF-β1 mRNA.
For treatment of pediatric inguinal hernia, we fabricated a device, i.e. so called "filling type pediatric hernia sac", which treats the problem from the abdominal cavity, through the abdominal and is a self-adaptive closer, using synthetic material. The device includes filling rack, self-adaptive umbrella support bar, bottom piece, outside pulling line and device fixing lines. The filling rack is composed of 2 concentric circles of 3.0 cm diameter with peripherally fixed together and can be pulled into the shapes of a ball or an olive. The supporting bar is structured of 3 pieces with 0.5 cm wide, 4.0 cm long, cross-fixed on top of the filling rack. The bottom piece is in a circular structure with a diameter of 3.0 cm, and it is connected to the filling rack bottom. Adjust positioning stay outside the fixed on the top of the device are connected at one end, and the other end free through filling the top frame connected with the bottom slice of central fixation. By using this device, we treated 37 pediatric inguinal hernia cases with 38 side-inguinal hernia successfully. The mean duration of post-operation follow-ups was 14.6±5.89 months, without hernia recurrence, obvious scar and hard sections of inguinal region. This device could provide a convenient, safe and effective plugging technology for children's pediatric hernia.
Objective To investigate the influence of enamel matrix proteins (EMPs) on the attachment, prol iferation and pre-mRNA of type I collagen synthesis of cultured human dermal fibroblast cells. Methods Human dermal fibroblast cells were obtained from human acrobystia and cultured in DMEM medium with 10% FBS. The 3rd to 6th passage cells were used. Ninety-six-well plates and 6-well plates were pre-coated with different concentrations of EMPs (50, 100, 150 and200 μg/ mL). ① The cell attachment experiment: 0.2 mL cells suspension at the concentration of 1 × 106/mL was added to the pre-coated 96-well plates as the experimental groups (groups A, B, C and D based on different concentrations of EMPs). At 1.5, 3.0, and 4.5 hours after inoculation, the attached cells were measured by MTT method. ② The cell prol iferation experiment: 0.2 mL cells suspension at the concentration of 5 × 104/mL was added to the pre-coated 96-well plates as the experimental groups (groups A1, B1, C1 and D1 based on the different concentrations of EMPs). At 2, 4, 6 and 8 days after inoculation, the cells were measured by MTT method. ③ The synthesis experiment of pre-mRNA: 2 mL cells at the concentration of 1 × 106/mL was added to the pre-coated 6-well plates as the experimental groups (groups A2, B2, C2 and D2 based on different concentrations of EMPs). At 5 days after inoculation, the synthesis of pre-mRNA was measured by RT-PCR method. Human dermal fibroblast cells were added to the un-coated plates as the control groups. Results ① The cell attachment experiment: There were significant differences in attachment cells between the control group, group A and the groups B, C and D (P lt; 0.05). There were no significant difference between group A and control group (P lt; 0.05). ② The cell prol iferation experiment: At 2 days, there were no significant differences in absorbance between the control group and the experimental groups (P gt; 0.05); at 4 days and 6 days, the absorbance of groups B1 (0.598 ± 0.020 and 0.639 ± 0.016 ), C1 (0.582 ± 0.017 and 0.641 ± 0.020) and D1 (0.574 ± 0.021and 0.635 ± 0.021) was significantly higher than that of the control group (0.548 ± 0.021 and 0.605 ± 0.019, P lt; 0.05); at 8 days, the absorbance of group B1 (0.629 ± 0.012) and group C1 (0.631 ± 0.014) was significantly higher than that of the control group (0.606 ± 0.031, P lt; 0.05). ③ The synthesis experiment of pre-mRNA: The synthesis of type I collage pre-mRNA of groups B2, C2 and D2 was significantly higher than that of the control group. Conclusion EMPs stimulate human dermal fibroblast cell attachment, prol iferation and synthesis of type I collage pre-mRNA, and its maximal effect can be achieved at the concentration of 100 μg /mL.
Objective To investigate the inhibition effect of silence of a disintegrin and metalloproteinase 17 (ADAM17) gene on proliferation and apoptosis of HT29 colon cancer cells and its possible mechanism. Methods HT29cells were divided into 3 groups: cells of interference group were transfected with recombinant lentivirus vector, cells of negative control group were transfected with negative recombinant lentivirus vector, and cells of blank control group were treated with PBS. The expression of ADAM17 mRNA was detected by real-time PCR, the expressions of ADAM17 protein, caspase3, protein kinase B (Akt), glycogen synthase kinase-3β (GSK3β), phospho-protein kinase B (P-Akt), phospho-glycogen synthase kinase-3β (P-GSK3β) protein were detected by Western blot method, the cell proliferation was detected by MTT assay, and the apoptosis rate was detected by Annexin V-FITC/PI cell death detection kit. Results Compared with the control group and the negative control group, the interference group was related to low expressions of ADAM17 mRNA and its protein, low optical density value at the same time point (24, 48, and 72 h), high apoptosis rate, high expression level of caspase3 protein, but low expression levels of P-Akt and P-GSK3β protein (P<0.05). Conclusion Silent ADAM17 gene could significantly induces apoptosis and inhibits the proliferation of HT29 cells, which maybe via inhibiting Akt/GSK3β signaling pathway.
【Abstract】ObjectiveTo explore the changes of colon motility of the rats in multiple organ dysfunction syndrome （MODS） induced bacterial peritonitis and the effects of IL6, TNFα and induce nitricoxide synthase (iNOS) on colon motility. MethodsWistar rats were divided into two groups, which were the control group and the MODS group. The number of stool, the amplitude changes of circular smooth muscle strip, the length of smooth muscle cell, and the changes of serum NO in two groups were observed. The expressions of IL6, TNFα and iNOS protein and IL6 mRNA, TNFα mRNA and iNOS mRNA in distal colon were investigated by using immunohistochemical methods and RTPCR. ResultsThe numbers of stool and the amplitude in the MODS group were lower than those of the control group （P＜0.05）. The expressions of IL6, TNFα and iNOS were negative in the control group, while they were positive in the MODS group. IL6 mRNA,TNFα mRNA and iNOS mRNA were negative expression in the control group, but they were positive expression in the MODS group. The concentration of serum NO and the length of smooth muscle cells in the MODS group were higher than those of the control group (P＜0.01). ConclusionColon motor dysfunction of the rats is related to the iNOS, IL6 and TNFα.