Tumour necrosis factor stimulated gene 6 intrinsically regulates PD-L1 expressions in breast cancer cells,leading to modulation of tumour microenvironment

Recent studies have shown that tumour cells express tumour necrosis factor-inducible gene 6 (TSG-6) and its protein, which is known to play a key role in regulating excessive immune responses and proliferation and growth of mesenchymal stem cells (MSCs). It has not been confirmed whether the inhibition of TSG-6 for tumour cells can suppress tumour cell growth and regulate the activation of immune cells in the tumour microenvironment (TME). TSG-6-specific small interfering RNA was transfected into canine and human breast cancer cells (CIPp, CIPm and BT-20). TSG-6-down-regulated (siTSG-6) cells showed decreased cell proliferation, migration, and invasion abilities. Decreased mRNA expressions of NF-κB, STAT3 and Sox2, confirming that TSG-6 is an upper factor governing tumour growth and metastasis. Notably, siTSG-6 cells showed significantly decreased expression levels of CD44 and PD-L1. Direct and indirect co-culture of canine peripheral blood mononuclear cells (cPBMCs) and the siTSG-6 cells showed significant activation in M1 type macrophages and cytotoxic T cells. They also showed a tendency to decrease in the expression of CTLA-4 and increase in the expression of PD-1. In conclusion, this study suggests that the down-regulation of TSG-6 in breast cancer cells could not only suppress tumour growth and metastasis, and but also regulate TME. Since modulation of immune checkpoint proteins occurs in both tumour cells and immune cells, inhibiting TSG-6 and its protein within the TME could be novel therapeutic target for anticancer treatment.     

DA-6对野大麦幼苗光合作用和生长的影响

研究了不同浓度的植物生长促进剂--DA-6对野大麦Hordeum brevisubulatum幼苗光合作用和生长的影响.在二叶期喷施较低质量浓度(5 mg/L,10 mg/L)的DA-6,可显著促进幼苗生长和叶绿素含量的提高;四叶期喷施较高质量浓度(20 mg/L,30 mg/L, 40 mg/L)的DA-6,可显著提高野大麦幼苗的生物量,其中20 mg/L和30 mg/L的处理,幼苗生物量分别提高55.1%和59.6%;光合作用能力分别提高12.8%和18.1%.     

结核分枝杆菌分泌蛋白ESAT-6、CFP10基因的克隆、鉴定及其原核表达

以人型结核分枝杆菌 H37RV株基因组 DNA为模板 ,应用 PCR法对 ESAT- 6和 CFP10基因进行扩增 ,产物经纯化后与载体 PMD18- T连接、转化及酶切鉴定 ,亚克隆到原核表达载体 PGEX- 6 P- 1,构建原核重组表达质粒 ,转化入大肠杆菌 BL2 1中 ,以 1mmol/L IPTG诱导 ,进行 SDS- PAGE电泳。结果表明 ,ESAT- 6和 CFP10基因表达的融合蛋白相对分子质量分别为 32 0 0 0和 36 0 0 0 ,与实测相符。重组结核杆菌分泌蛋白 ESAT- 6和 CFP10的成功表达为结核病诊断及重组疫苗的构建打下了基础     

Involvement of the ERK1/2 MAPK pathway in insulin-induced S6K1 activation in avian cells

In mammals, insulin regulates S6K1, a key enzyme involved in the control of protein synthesis, via the well-documented phosphoinositide-3'kinase (PI3K) pathway. Conversely, S6K1 is activated by insulin in avian muscle despite the relative insulin insensitivity of the PI3K pathway in this tissue. Mitogen-activated protein kinase (MAPK) cascade is another insulin sensitive pathway. The aim of this study was to explore the potential involvement of the ERK1/2 MAPK pathway in the control of p70 S6 kinase (S6K1) in avian species. Firstly, we characterized ERK1/2 MAPK in various chicken tissues. ERK2 was the only isoform detected in avian species whatever the tissue studied. We also showed that ERK2 is activated in vivo by insulin in chicken muscle. The regulation and the role of ERK2 in insulin signaling were next investigated in chicken hepatoma cells (LMH) and primary myoblasts. Insulin stimulation led to ERK2 and S6K1 phosphorylation, and concomitantly increased kinase activity. U0126, an inhibitor of the ERK MAPK pathway, completely abolished insulin-induced S6K1 phosphorylation and activity in chicken myoblasts, whereas its effect was only partial in LMH cells. In conclusion, these results show that ERK1/2 MAPK is involved in the control of S6K1 by insulin in chicken cells, particularly myoblasts.     

生长激素释放肽-6缓释微球的制备及其对动物生长的影响

以生物降解材料乳酸乙醇酸共聚物(PLGA,LA-GA 8515)为载体,采用复乳-液中蒸发法制备含生长激素释放肽-6的乳酸乙醇酸共聚物微球,并通过体外药物释放试验评价载药微球的释药特征.结果得到收率、粒径大小和分布及含药量均满意的微球,其中包封率为100%,平均体积径为3.45μm,收率为79%.在37℃、pH7.0的生理等渗缓冲液中,首日释药19.19%,其后以平均日释药3.23%的速度释药25 d.小鼠肌肉注射微球30 d后,累积增重比注射生长激素释放肽-6、生理盐水分别高22.56%(P<0.05)和53.17%(P<0.01).结果表明,生长激素释放肽-6乳酸乙醇酸共聚物微球有望发展成为新型长效制剂.     

6-BA和保水剂对匍匐翦股颖种茎耐贮性的影响

用不同浓度的6-BA、保水剂和6-BA＋保水剂配合处理粤选1号匍匐翦股颖种茎,以清水做对照（CK）,以探讨它们对匍匐翦股颖种茎采后贮藏2～16d的影响。结果表明：经6-BA、保水剂处理后后种茎保水能力提高,但保水剂处理易造成种茎贮藏期间萌动。进一步试验表明,贮藏期问7．5和10mg／L6-BA以及7．5mg／L6-BA＋2g／L保水剂处理种茎出苗势、成活率均有所提高,尤其是7．5和10mg／L6-BA处理的种茎在贮藏第16d成活率仍高速909,6以上,与对照相比差异显著（P〈0.05）;各处理对种茎萌发后的地上和地下生长均有一定促进作用;单施保水剂处理种茎在贮藏期间易萌动,不利于种茎贮藏。     

Adaptive evolutionary expansion of the ribonuclease 6 in Rodentia

Ribonuclease 6 (RNase6 or RNase K6) is a protein that belongs to a superfamily thought to be the sole verte‐brate‐specific enzyme known for a wide range of physiological functions, including digestion, cytotoxicity, angiogenesis, male reproduction and host defense. In our study, 51 functional genes and 11 pseudogenes were identified from 27 Rodentia species. Intriguingly, in the 3 main lineages of rodents there were multiple RNase6s identified in all species of Ctenohystrica, whereas only a single RNase6 was observed in other Rodentia species examined except for 2 species in the mouse‐related clade. The evolutionary scenario of “birth (gene duplication) and death (gene deactivation)” and gene sorting have been demonstrated in Ctenohystrica. In addition, bursts of positive selection, diversification of isoelectric point and positive net charge have been identified in Ctenohystrica, especially at two key sites that are involved in antimicrobial function. Site Trp30 has undergone positive selection and Ile45 has changed into other residues in Group B and Group C of the Ctenohystrica. Our results demonstrated a complex and intriguing evolutionary pattern of rodent RNase6, and indicated that functional modification may have occurred, which establishes an important theoretical foundation for future functional assays in rodent RNase6.     

仔猪雷极氏普罗菲登斯菌的分离鉴定及系统进化分析

为了解猪雷极氏普罗菲登斯菌(P.rettgeri)的生物学特性、致病性及16SrRNA基因系统进化关系,从发生严重腹泻、血便的哺乳仔猪群的内脏器官中分离到1株病原菌,根据形态特征、培养特性、生化特性、16SrRNA基因序列分析鉴定为P.rettgeri。小鼠攻毒试验证实,该分离菌株对试验小鼠有较强致病力,哺乳仔猪回归试验可复制出与临床上相似的典型症状,并从发病死亡小鼠及哺乳仔猪中分离到攻毒菌株。系统进化分析表明,分离菌株与雷极氏普罗菲登斯菌系统进化关系最为密切,其16SrRNA基因序列与雷极氏普罗菲登斯菌代表菌株的同源性在97.3%~99.6%之间。药敏试验结果表明,分离菌株对头孢哌酮钠、头孢噻肟钠、头孢曲松钠、头孢唑啉、头孢呋肟、头孢吡肟、阿莫西林、链霉素、氨曲南、诺氟沙星、左氟沙星、恩诺沙星、卡那霉素、氨苄青霉素、米诺环素、链霉素、阿米卡星等多数药物敏感。首次报道了雷极氏普罗菲登斯菌可以引起哺乳仔猪严重腹泻,提示在仔猪腹泻中应注意该菌感染的诊断、监测和防控。     

绵羊DUSP6基因多态性及其与产羔数关联分析

文章旨在探究绵羊DUSP6基因g.125589716G>A、g.125587728C>T、g.125589714C>T、g.125589006G>A四个位点多态性及其与产羔数之间的关系,以期找到与绵羊高繁殖力相关的分子标记。利用全基因组重测序结合Sequenom MassARRAY誖SNP技术对多羔绵羊品种(小尾寒羊、湖羊、策勒黑羊)和单羔绵羊品种(滩羊、苏尼特羊、萨福克羊、草原型藏羊)DUSP6基因上述4个多态位点进行检测,并与小尾寒羊产羔数进行关联分析。结果表明:绵羊g.125589716G>A位点在多羔绵羊品种中存在AA、GA、GG三种基因型,在单羔绵羊品种中存在GA、GG两种基因型;g.125587728C>T、g.125589714C>T位点在单、多羔绵羊品种中均存在CC、CT、TT三种基因型;g.125589006G>A位点在单、多羔绵羊群体中均只存在GA、GG两种基因型。绵羊DUSP6基因g.125589716G>A位点基因型频率在单、多羔绵羊品种之间差异显著(P<0.05),等位基因频率在单、多羔绵羊品种间差异极显著(P<0.01);g.125587728C>T、g.125589714C>T位点基因型频率和等位基因频率在单、多羔绵羊品种间差异均达极显著水平(P<0.01);g.125589006G>A位点基因型频率和等位基因频率在单、多羔绵羊品种间差异不显著(P>0.05)。关联分析表明,DUSP6基因各SNPs多态性与小尾寒羊不同胎次产羔数之间无显著关联(P>0.05)。综上说明,绵羊DUSP6基因g.125589716G>A、g.125587728C>T、g.125589714C>T以及g.125589006G>A等4个位点的多态性与小尾寒羊各胎次产羔数之间均无显著关联(P>0.05),不适用于小尾寒羊多羔性状选育。