Identification and establishment of sorting method for isolating CD11b+ myeloid cells in human hepatic metastases from colorectal cancer

AIM: To establish a method for obtaining specific cells in solid tumor tissue by sorting of CD11b⁺ myeloid cells in hepatic metastases from colorectal cancer.METHODS: Tumor tissues were prepared into single cell suspension by mechanical method combined with enzyme digestion, and then the CD11b⁺ myeloid cells were isolated by flow cytometry. The sorted cells were identified by immunocytochemistry. The viability and morphologiy of the sorted cells were evaluated by Giemsa and Typan blue staining. The cell purity was evaluated by flow cytometry.RESULTS: Sufficient numbers of CD11b⁺ cells with high purity were isolated by sorting with flow cytometry from the single cell suspension prepared by mechanical and enzyme digestion. The purity of the cells was confirmed by statistical analysis (P<0.05). The positive rates of the cells before and after sorting were significantly different (P<0.01). The positive cells were verified by immunocytochemical method. Meanwhile, the sorted cells had complete morphology and good activity.CONCLUSION: The CD11b⁺ myeloid cells in solid tumor tissue can be isolated by flow cytometry from the machine-enzyme digestion suspension with high purity, good activity and complete morphology.     

Advances on studies of miR-21 in vascular endothelial function and angiogenesis

MicroRNAs (miRNAs)are a class of non-coding, endogenous, single-stranded small RNA molecules composed of 19~25 nucleotides. miRNAs are widely involved in the process of human life activities. Recent studies have shown that part of miRNAs regulate the vascular endothelial function and angiogenesis. High expression of miRNA-21 is found to play important roles in the cell proliferation, cell apoptosis, cell growth and death of vascular endothelial cells. This review will focus on the recent progress related to miRNAs in vascular endothelial function and angiogenesis, providing a new insight in cardiovascular disease prevention, clinical diagnosis, prognosis and target therapeutics.     

Effect of eleutheroside B/E on proliferation and expression of PPARγ and TGF-β1 in HBZY-1 cells cultured with high glucose

AIM: To explore the effects and mechanism of eleutheroside (ETS) B or E on the proliferation of HBZY-1 cells treated with high glucose. METHODS: The HBZY-1 cells were cultured under high glucose condition. The 4th generation of HBZY-1 cells was used for determining the optimal cell density, which was consistent with the growth regulation curve of the cells. The cells were divided into 6 groups: low glucose (LG) group, high glucose (HG) group, high glucose plus ETS-B/E (low dose, medium dose and high dose) groups, and high glucose plus losartan (LTG) group. After all cells were treated with the corresponding drugs at 24 h, 48 h and 72 h, the inhibitory rate of the proliferation was measured, and the expression of TGF-β1 and PPARγ was detected by immunocytochemistry and Western blotting. RESULTS: The best cell density was 2 000 cells/well, which was complied with the basic rules of the cell growth, and high glucose significantly promoted the HBZY-1 cell proliferation. At each time point, the inhibitory effects of ETS-B/E were significantly different between HG group and LTG group on the proliferation of the HBZY-1 cells (P<0.05). The expression of TGF-β1 was significantly inhibited, and the expression of PPARγ was significantly promoted by ETS-B/E (P<0.05). ETS-E showed stronger effect than ETS-B (P<0.05) in a concentration- and time-dependent manner. CONCLUSION: ETS-B/E significantly inhibits the proliferation of HBZY-1 cells under high glucose condition by decreasing TGF-β1 expression and promoting PPARγ expression.     

Gonadal morphogenesis and sex differentiation in cultured chub mackerel,Scomber japonicus

The timing of primordial germ‐cell (PGC) migration with regard to the gonadal anlagen, gonad formation and sex differentiation was examined histologically in the chub mackerel (Scomber japonicus) at 5–190 days post hatching (dph). At 5 dph, PGCs appeared on the peritoneal epithelium surface or in the mesentery, on the dorsal side of the abdominal cavity. By 10 dph, stromal cells around the PGCs proliferated. The gonadal primordium was formed by 15 dph. The gonadosomatic index was 0.01% at 30 dph and increased thereafter (0.32% in females and 0.04% in males at 160 dph). Ovarian differentiation occurred at 30–40 dph, indicated by ovarian cavity formation (elongation and fusion of the upper and lower ovarian edges). Meiosis was subsequently initiated. A few meiotic oocytes surrounded the cavity at 50 dph; most were in the perinucleolus stage at 60 dph and attained a diameter of 60–70 μm at 190 dph. Testicular differentiation occurred at 30 dph, indicated by the formation of the sperm duct primordium. Spermatogonia gradually proliferated, developing into spermatocytes at the chromatin–nucleolus stage (after 90 dph) and subsequently into spermatids and spermatozoa (160 dph). These data could aid the development of seeding and cell‐engineering technologies for scombrid fish.     

铅和（或）镉对体外培养肾细胞的毒性损伤

在培养液中加入不同浓度的Cd（10、20、40μmol／L）、Pb（10、20、40μmol／L）和Pb-Cd联合（分别为5、10、20μmol／L），来探讨铅镉对肾细胞的作用及可能机制。在显微镜下观察细胞形态、测定细胞凋亡率和细胞周期及上清液脂质过氧化指标。结果表明：20μmol／L以上染毒组细胞皱缩、体积变小，表面有细胞碎片，甚至呈泡沫状或树枝状；铅、镉单独和联合染毒组GSH—Px、SOD活性随染毒剂量的增加，呈逐渐下降趋势（P〈0．05）．而MDA含量和凋亡率则呈升高趋势（P〈0．05），并存在剂量和时间效应。镉和铅也能使细胞周期停滞。Pb、Cd联合可加剧细胞损伤。     

厚颌鲂外周血细胞形态及细胞化学特征研究

采用血细胞常规染色和细胞化学染色对厚颌鲂(Megalobrama pellegrini)外周血细胞形态及细胞化学特征进行观察。结果显示:厚颌鲂外周血细胞可分为红细胞、血栓细胞、单核细胞、淋巴细胞和三种类型粒细胞。红细胞有幼稚、成熟和衰老三种形态,其中幼稚和衰老红细胞较少见;血栓细胞数量最多,其次是淋巴细胞。粒细胞中Ⅰ型和Ⅱ型较常见,Ⅲ型粒细胞少有。细胞化学染色显示幼红细胞、血栓细胞和淋巴细胞均呈ACP弱阳性;单核细胞呈ACP和ANAE阳性;Ⅰ型粒细胞是嗜中性粒细胞,POX、ACP和ANAE阳性;Ⅱ型粒细胞是PAS-GL,PAS强阳性和ACP阳性;Ⅲ型粒细胞是Ⅰ型粒细胞的晚期形态。厚颌鲂的PAS-GL可能是嗜酸性/嗜碱性粒细胞,而不是嗜碱性粒细胞的前体。     

激素对奶牛乳腺上皮细胞乳脂肪合成的调控作用

乳脂肪是高质量的天然脂肪，其可为人类提供营养和能量，在各种膳食脂肪和油类中，是最容易被消化吸收的。乳脂肪是在乳腺中由从头合成或外源摄取的脂肪酸与甘油酯化形成的一种脂类物质，其含量的高低关系着牛奶品质的优劣和乳制品的加工特性。在奶牛的泌乳周期中，乳腺泌乳功能受多种因素影响，其中内分泌腺分泌的多种激素对奶牛乳腺上皮细胞（BMECs）乳脂的合成具有积极的调控作用。综上所述，作者介绍了氢化可的松、催乳素、胰岛素和生长激素4种泌乳相关激素对BMECs乳脂肪合成的调控机理，即从乳脂合成适宜的激素添加量、激素对乳脂球形态的影响方面初步阐释其调控作用，并从乳脂合成的关键酶及转录因子、激素对乳脂合成相关基因表达量方面深入阐释其作用机理，旨在为研究泌乳相关激素对奶牛乳腺内乳脂肪合成的调控机理提供参考。     

超氧化物歧化酶模拟物对氧化应激IPEC-J2细胞抗氧化性能的影响

试验旨在探究在H₂O₂诱导的氧化应激状态下，超氧化物歧化酶模拟物（SODm）对仔猪空肠上皮细胞系（IPEC-J2）的保护作用。利用MTT法筛选出构建氧化应激模型H₂O₂的适宜浓度；将IPEC-J2细胞分别用0、0.05、0.5、5、50、500、2 000 U/mL SODm进行培养，利用MTT法分别在2、4、8、12 h时测定各组细胞存活率，筛选出SODm作用的适宜浓度和时间；根据构建的氧化应激模型和SODm适宜浓度和时间，将IPEC-J2细胞随机分为空白组、模型组、SODm处理组（SODm_0.5、SODm₅、SODm₅₀）和超氧化物歧化酶（SOD）处理组（SOD_0.5、SOD₅、SOD₅₀），分别测定各组细胞存活率、活性氧（ROS）含量及细胞内SOD、谷胱甘肽过氧化物酶（GSH-Px）活性及丙二醛（MDA）含量和总抗氧化能力（T-AOC）。结果表明：①H₂O₂构建细胞氧化应激模型的适宜浓度是1.0 mmol/mL。②当不同浓度SODm分别作用4、8、12 h时，0.5~50 U/mL SODm组细胞存活率显著高于空白组（P<0.05）；且8 h时存活率最高，在12 h时处于下降趋势。③除空白组外，SODm₅和SOD₅预处理的IPEC-J2存活率显著高于其他组（P<0.05）；且SODm和SOD组中细胞ROS含量显著低于模型组（P<0.05）；模型组与空白组相比，均显著降低了SOD、GSH-Px活性和T-AOC水平，提高了MDA含量（P<0.05）；与模型组相比，所有SODm和SOD处理组均显著提高了SOD、GSH-Px活性和T-AOC水平，降低了MDA含量（P<0.05），同时SODm₅₀组T-AOC水平显著低于SOD₅₀组（P<0.05）。综上，SODm对H₂O₂诱导氧化损伤状态下IPEC-J2具有保护作用。     

Effect of xeroderma pigmentosum group D gene on proliferation of human umbilical arterial smooth muscle cells induced by Ox-LDL

AIM: To investigate the effects of xeroderma pigmentosum group D (XPD) gene on the proliferation of human umbilical arterial smooth muscle cells (HUASMCs) induced by oxidized low-density lipoprotein (Ox-LDL). METHODS: The recombinant plasmid pEGFP-N2/XPD was transfected into HUASMCs by liposome. The cells were divided into blank control group, pEGFP-N2 group, pEGFP-N2/XPD group, Ox-LDL group, Ox-LDL+pEGFP-N2 group and Ox-LDL+pEGFP-N2/XPD group. The proliferation rate of the cells was detected by MTT and EdU assays. The apoptotic rate and cell cycle distribution were analyzed by flow cytometry. The protein levels of XPD, caspase-3, Bcl-2 and Bax were determined by Western blot. RESULTS: Compared with blank control group, the expression of XPD was increased in pEGFP-N2/XPD group (P<0.05). According to the results of MTT and EdU assays, the cell proliferation in pEGFP-N2/XPD group was reduced compared with blank control group (P<0.05). Compared with Ox-LDL group, the cell proliferation in Ox-LDL+pEGFP-N2/XPD group was significantly inhibited (P<0.05). According to the results of flow cytometry, the cell proportion of S phase decreased and the G₀/G₁-phase cell proportion increased significantly in pEGFP-N2/XPD group and Ox-LDL+pEGFP-N2/XPD group compared with blank control group and Ox-LDL group, repectively (P<0.05). Compared with blank control group and Ox-LDL group, the protein level of Bcl-2 decreased and the protein levels of Bax and cleaved caspase-3 increased in pEGFP-N2/XPD group and Ox-LDL+pEGFP-N2/XPD group, respectively (P<0.05). CONCLUSION: XPD inhibits the proliferation of HUASMCs and promotes their apoptosis, and reduces the promoting effect of Ox-LDL on the proliferation of HUVSMCs. XPD may be the target for treatment of atherosclerosis.     

蜂王浆对尾静脉注射4T1细胞小鼠抗氧化能力的影响

探讨蜂王浆对尾静脉注射4T1细胞小鼠的抗氧化和免疫力的影响.将4T1细胞或其与蜂王浆的混合物通过尾静脉注射入小鼠体内,ELISA法检测蜂王浆对小鼠血清、肝脏和肾脏中与抗氧化能力相关的指标及血清中和免疫力相关细胞因子水平的影响.结果表明,和模型对照组相比,蜂王浆(50、100 mg·mL~(-1))可以显著提高小鼠血清内T-AOC、IFN-α(P0.05)水平,降低IL-4的水平(P0.05);蜂王浆剂量为50 mg·m L~(-1)时,IFN-γ的含量显著增加(P0.05).蜂王浆(100 mg·mL~(-1))显著提高肾脏中T-AOC的水平(P0.05).可见,蜂王浆可以改善尾静脉注射4T1细胞小鼠的抗氧化能力和免疫力.