Ultrastructural impairment of islet microvascular endothelial cells in STZ-induced type 1 diabetic mice

AIM: To investigate the ultrastructural changes of islet microvascular endothelial cells in STZ-induced type 1 diabetic mice. METHODS: BALB/c mice were randomly divided into diabetic group and control group. The expression of insulin and platelet-endothelial cell adhesion molecule-1 (CD31) in islet microvessels was detected by immunohistochemical staining. The ultrastructural changes of islet β cells and islet microvessels were observed under transmission electron microscope. RESULTS: Compared with control group, the number of islet β cells, ratio of β cells/α cells, average number of secretory granules in β cells and insulin expression area per islet in diabetic group were significantly decreased (P<0.01). Besides, diabetic group had fewer microvessels with lower expression of CD31 (P<0.01). Mitochondria in islet microvascular endothelial cells and pericytes in diabetic group were swelling. The basement membrane of islet microvessels became thicker in diabetic group (P<0.01). CONCLUSION: Islet microvascular endothelial cells were impaired in type 1 diabetic mice.     

Vitamin C enhanced myocardial differentiation of dedifferentiated fat cells

AIM: In order to observe the myocardial differentiation capacity of the dedifferentiated fat (DFAT) cells treated with vitamin C in vitro. METHODS: DFAT cells were dedifferentiated from the mature rat adipocytes with ceiling adherent culture. The DFAT cells of passage 3 were used in the study. Vitamin C and/or neonatal rat heart tissue lysate were added into the culture medium to induce myocardial differentiation for 3 weeks. The cell morphology was observed under microscope. The myocardial-specific markers, such as cTnT, GATA-4 and NKx2.5, were examined by the methods of immunofluorescence, PCR and Western blot. RESULTS: Mature rat adipocytes dedifferentiated into fibroblast-like DFAT cells after ceiling adherent culture. The DFAT cells spontaneously differentiated into cardiomyocyte-like cells under normal culture condition with a low incidence. After treated with neonatal rat heart cell lysate, the DFAT cells became cardiomyocyte-like cells that had bigger size, longer shape and myotubule-structure. The expression of cTnT, GATA-4 and NKx2.5 was remarkably increased at both mRNA and protein levels as compared with the normal cultured DFAT cells. The expression of cTnT, GATA-4 and NKx2.5 was further increased in DFAT cells after treating with vitamin C. No spontaneous beating cell was observed. CONCLUSION: Vitamin C enhances the differentiation of DFAT cells into cardiomyocyte-like cells.     

Model construction of rat coronary artery smooth muscle cell endoplasmic reticulum stress induced by thapsigargin

AIM: To investigate the primary culture method for coronary artery smooth muscle cells (CASMCs), and to establish the endoplasmic reticulum stress (ERS) model in CASMCs of SD rats. METHODS: CASMCs were cultured by tissue explant method. The morphological characteristics were observed under optical microscope. The marker proteins of CASMCs, including α-SMA and SM-MHC, were identified by immunofluorescence technique. The protein expression levels of BiP and CHOP, the marker molecules of ERS, were determined by Western blot. RESULTS: The spindle-shaped CASMCs climbed out from the edge of coronary artery tissues after 6 d, and formed the typical "hill and valley" growth pattern of CASMCs at 9~10 d. The result of immunofluorescence technique showed that α-SMA and SM-MHC were positively expressed. The results of Western blot showed that the protein expression of BiP and CHOP in TG (1 and 2 μmol/L) treatment groups was increased compared with control group. Compared with control group, the protein expression of BiP and CHOP was significantly increased after 1 μmol/L TG treatment for 24 and 48 h. CONCLUSION: CASMCs can be successfully cultured by tissue explant method. ERS model of CASMCs was established by 1 μmol/L TG treatment for 24 h.     

Periplaneta americana peptide decreases apoptosis of pig-ovary granulosa cells induced by H2O2 through FoxO1

Oxidative stress can induce apoptosis of granulosa cells and lead to follicular atresia, thereby reducing the number of pigs giving birth. The aim of this study was to investigate the protective effect of Periplaneta americana peptide (PAP) on the apoptosis of the granulosa cells of pig ovaries (PGCs) induced by hydrogen peroxide (H₂O₂) via FoxO1. PGCs were treated with H₂O₂ to establish a cell apoptosis model. Cell viability was measured using the cell counting kit-8 (CCK-8) assay, and cell apoptosis was detected using flow cytometry. The malondialdehyde (MDA) level and nitric oxide (NO) content were detected to reflect the oxidative stress. Western blotting, qRT-PCR and overexpression were undertaken to determine the expression of FoxO1 and caspase-3, and immunofluorescence was used to detect FoxO1 in the nucleus and cytoplasm. PGCs were treated with 100 μM H₂O₂ for 6 hr, which resulted in oxidative damage and apoptosis and an apoptosis rate for PGCs of 32.95%. Next, PGCs were treated with 400 μg/ml PAP for 24 hr to repair the apoptosis induced by H₂O₂. PAP improved cell viability in H₂O₂-stimulated PGCs, the increased MDA level and NO content caused by H₂O₂ stimulation were reversed and the apoptotic rate of PGCs was reduced. The qRT-PCR and Western blotting results indicated that PAP decreased the H₂O₂-induced apoptosis and the expression of FoxO1 and caspase-3 in PGCs. The effect of PAP was the same following FoxO1 overexpression. FoxO1 was expressed in the nucleus when stimulated by H₂O₂ or overexpression; however, it migrated to the cytoplasm following PAP treatment. PAP decreased the apoptosis of PGCs induced by H₂O₂ by regulating FoxO1 expression and nuclear translocation.     

猪lncRNA TCONS_00791383对骨骼肌卫星细胞增殖分化的影响

为了探究lncRNA TCONS_00791383对猪骨骼肌卫星细胞增殖和分化的影响。本研究利用qRT-PCR技术检测出生7 d内大白仔猪6种组织（心、脾、肺、肾、背肌和腿肌）以及猪骨骼肌卫星细胞增殖分化前后TCONS_00791383的表达水平；通过设计反义核苷酸（antisense oligonucleotides，ASO）片段在猪骨骼肌卫星细胞中对TCONS_00791383进行敲低，检测敲低TCONS_00791383之后增殖分化标志基因的表达量变化；通过trans （co-expression）对TCONS_00791383进行靶基因预测，使用DAVID对其进行GO富集和KEGG通路分析。结果显示，TCONS_00791383在猪心脏中表达量最高，在脾和肾组织中不表达。在骨骼肌卫星细胞从增殖到分化的过程中，TCONS_00791383的表达量逐渐上升，且在分化后30 h表达量达到最高。在使用ASO片段敲低TCONS_00791383之后，与对照组相比，在分化24 h，增殖标志基因Pax3、Pax7表达量显著或极显著降低（P<0.05，P<0.01），分化标志基因MyoG表达量极显著降低（P<0.01），在分化48 h，增殖标志基因Pax3表达量极显著降低（P<0.01），Pax7表达量显著降低（P<0.05），分化标志基因MyHC表达量显著降低（P<0.05）。预测得到的相关靶基因富集到AMPK、ATP等多个与骨骼肌卫星细胞增殖和分化过程相关的重要信号通路。本研究表明，lncRNA TCONS_00791383可能促进猪骨骼肌卫星细胞的增殖和分化。     

密集烤房不同循环风机烘烤烟叶的色素降解及感官质量

采用三速和单速循环风机,研究密集烤房不同循环风机对烤烟色素及感官质量的影响.结果表明:采用三速循环风机烤后烟叶的叶绿素a、叶绿素b含量分别为5.10、167.74 μg/g,分别为单速循环风机的72.11％和93.30％;叶黄素、β-胡萝卜素、类胡萝卜素含量分别为0.84、155.56、156.40 μg/g,分别为单...     

番茄绿果与橙果间果实颜色及主要色素含量的遗传研究

对番茄组合绿樱(绿果)×金珠1号(橙果)的6个世代遗传群体(P1、P2、F1、BC1、BC2和F2)进行果色性状、番茄红素含量、叶绿素含量和胡萝卜素含量等的遗传规律分析。结果表明:正反交F1的果色性状无明显差异,而色素含量存在显著差异;说明番茄果色性状受核基因控制,而色素含量遗传除受核基因控制外还可能存在胞质效应。采用多世代联合分析法的分析结果表明,番茄绿果与橙果间的果色性状符合2对加性主基因+加性-显性多基因(MX2-A-AD)遗传模型,其BC1、BC2和F2主基因遗传率分别为73.42%、78.25%和61.41%,多基因遗传率分别为22.87%、15.35%和34.94%,即果色性状遗传的主基因遗传力较强;叶绿素含量符合1对负向显性主基因+加性-显性多基因(MX1-AEND-AD)遗传模型,其BC1、BC2和F2主基因遗传率分别为0、1.73%和0.65%,多基因遗传率分别为45.47%、0和37.82%,即主基因遗传力在BC2群体中最高,多基因遗传力在BC1群体中最高;番茄红素含量与胡萝卜素含量均符合2对加性-显性-上位性主基因+加性-显性多基因(MX2-ADI-AD)遗传模型,其BC1、BC2和F2主基因遗传率分别为75.74%、1.79%、84.26%和61.53%、87.21%、81.05%,多基因遗传率分别为20.32%、74.12%、12.68%和0.68%、0、0,表明番茄红素含量和胡萝卜素含量的主基因遗传力较强。     

Inhibitory effect of poncirin on viability of gastric cancer cells and underlying mechanism

AIM: To explore the effect of poncirin on the growth of AGS gastric cancer cells and the underlying mechanism.METHODS: The effect of poncirin on AGS cell viability was measure by MTT assay. The cell cycle distribution and cell apoptosis were analyzed by flow cytometry. Nuclear staining with DAPI was used to reflect the morphological change of the AGS cells treated with poncirin. The protein levels of extrinsic apoptosis pathway-related proteins such as FasL, caspase-8, caspase-3 and PARP, and mitochondria-mediated intrinsic apoptosis pathway-associated proteins such as Bak, Bcl-xL, Bax and caspase-9 were determined by Western blot.RESULTS: Poncirin inhibited the viability of AGS gastric cancer cells in a time-and concentration-dependent manner (P<0.05). Poncirin induced accumulation of G₁ DNA content and significantly increased total apoptosis in the AGS cells. Nuclear staining showed a dose-dependent increase in the number of apoptotic cells after treated with poncirin.The protein level of FasL was upregulated in a dose-dependent manner by treatment with poncirin. Poncirin significantly activated caspase-8 and caspase-3. Moreover, poncirin significantly induced the cleavage of PARP in a dose-dependent manner (P<0.05). In addition, the protein levels of Bcl-xL, Bax and Bak were unchanged after treated with different doses of poncirin. Furthermore, caspase-9 was not activated by poncirin treatment in the AGS cells.CONCLUSION: Poncirin has the anti-cancer effect via extrinsic apoptosis pathway to inhibit the growth of AGS gastric cancer cells, possibly making it a therapeutic agent for human gastric cancer treatment.     

消化酶及胎牛血清对奶牛乳腺上皮细胞培养的影响

试验研究了不同浓度及组合消化酶对奶牛乳腺组织分离的影响以及不同FBS浓度培养基对奶牛乳腺上皮细胞纯化和生长的影响。试验结果表明,0.25%胰酶＋0.20%Ⅱ胶原酶对组织块分离效果最好,乳腺上皮细胞生长最多,5%与10%浓度FBS对奶牛乳腺上皮细胞纯化效果最好,5%与10%浓度FBS对细胞生长没有明显的差异（P〉0.05...