花生种子老化相关蛋白质的初步研究

本文以粤油 116花生(Arachis hypogaea L.)为材料,对不同处理种子的除子叶“种胚”(以下简称“种胚”)的蛋白质进行了研究.实验结果表明,当花生种子活力下降到一定程度时,其“种胚”内出现一种新蛋白质( pI6.2、MW 10 KD),随种子老化程度加深,含量逐渐增多.我们认为该蛋白质与花生种子老化存在着一定的相关关系,可作为该种子老化的标志.     

低温对氯化钠胁迫下蓝藻固氮活性的影响

低温加剧氯化钠对蓝藻固氮的抑制,营养液中氯化钠浓度增高时,抑制程度更甚.能源受限(暗处理和加抑制剂时的光合受抑,N_2和Ar的厌氧下呼吸代谢受阻)和氧下固氮酶受到伤害时,低温处理使氯化钠对蓝藻固氮的抑制进一步加剧.在能源和还原剂供应,合成固氨酶蛋白的物质基础(如CO_2和N_2的加合).光合作用正常进行的条件得到改善和保证,以及供应CO_2、外源蔗糖和氮氧加合时,低温加剧氯化钠对蓝藻固氮的抑制程度明显变小.     

UreB immunodominant epitope-specific CD8+ T-cell responses were beneficial in reducing gastric symptoms in Helicobacter pylori-infected individuals

Background and aims

Although Helicobacter pylori is recognized as an extracellular infection bacterium, it can lead to an increase in the number of CD8⁺ T cells after infection. At present, the characteristics of H. pylori antigen-specific CD8⁺ T cells and the epitope response have not been elucidated. This study was focused on putative protective antigen UreB to detect specific CD8⁺ T-cell responses in vitro and screen for predominant response epitopes.

Methods

The PBMCs collected from H. pylori-infected individuals were stimulated by UreB peptide pools in vitro to identify the immunodominant CD8⁺ T-cell epitopes. Furthermore, their HLA restriction characteristics were detected accordingly by NGS. Finally, the relationship between immunodominant responses and appearance of gastric symptoms after H. pylori infection was conducted.

Results

UreB-specific CD8⁺ T-cell responses were detected in H. pylori-infected individuals. Three of UreB dominant epitopes (A-2 (UreB_443–451: GVKPNMIIK), B-4 (UreB_420–428: SEYVGSVEV), and C-1 (UreB_5–13: SRKEYVSMY)) were firstly identified and mainly presented by HLA-A*1101, HLA-B*4001 and HLA-C*0702 alleles, respectively. C-1 responses were mostly occurred in H. pylori-infected subjects without gastric symptoms and may alleviate the degree of gastric inflammation.

Conclusions

The UreB dominant epitope-specific CD8⁺ T-cell response was closely related to the gastric symptoms after H. pylori infection, and the C-1 (UreB_5-13) dominant peptides may be protective epitopes.     

福州地区桑白蚧发生动态和药剂防治试验

桑白蚧在福州地区一年发生4代．以雌成虫在寄主枝干上越冬．越冬代(第4代)一雌虫产卵量多的达278粒．少的36粒,平均171粒,比第2代产卵量多2．6倍．比第3代多4．5倍．药剂防治试验结果,在2龄幼蚧高峰期,用25％扑虱灵可温性粉剂1500倍液,40％氧化乐果乳油800-1000倍液和95%机油乳剂50—100倍液喷雾．防治效果可达90％左右．用25％扑虱是可湿性粉剂1000-1500倍液．喷酒幼蚕触杀试验和喷洒桑叶喂蚕胃毒试验结果．对幼蚕安全．用扑虱灵防治桑树上的桑白蚧,对养蚕业无不良影响。     

Bisphenol S induces oxidative stress-mediated impairment of testosterone synthesis by inhibiting the Nrf2/HO-1 signaling pathway

Bisphenol S (BPS) is an environmental endocrine disruptor widely used in industrial production. BPS induces oxidative stress and exhibits male reproductive toxicity in mice, but the mechanisms by which BPS impairs steroid hormone synthesis are not fully understood. Nuclear factor erythroid 2-related factor 2(Nrf2)/HO-1 signaling is a key pathway in improving cellular antioxidant defense capacities. Therefore, this study explored the effects of exposure to BPS on testosterone synthesis in adult male mice and its mechanisms with regard to the Nrf2/HO-1 signaling pathway. Adult male C57BL/6 mice were orally exposed to BPS (2, 20, and 200 mg/kg BW) with sesame oil as a vehicle (0.1 ml/10 g BW) per day for 28 consecutive days. The results showed that compared with the control group, serum testosterone levels were substantially reduced in the 20 and 200 mg/kg BPS treatment groups, and testicular testosterone levels were reduced in all BPS treatment groups. These changes were accompanied by a prominent decrease in the expression levels of testosterone synthesis-related enzymes (STAR, CYP11A1, CYP17A1, HSD3B1, and HSD17B3) in the mouse testis. In addition, BPS induced oxidative stress in the testis by upregulating the messenger RNA and protein levels of Keap1 and downregulating the levels of Nrf2, HO-1, and downstream antioxidant enzymes (CAT, SOD1, and Gpx4). In summary, our results indicate that exposure of adult male mice to BPS can inhibit Nrf2/HO-1 signaling and antioxidant enzyme activity, which induces oxidative stress and thereby may impair testosterone synthesis in testicular tissues, leading to reproductive damage.     

恒河猴(Macaca mulatta)卵细胞采集及体外受精的研究

对13头成年母猴,用黄体酮 前列腺索(PGF_(2a))处理,诱发月经周期,并用三种不同的处理方案,注射外激性促性腺激素诱发等超数排卵。对171个大滤泡(直径≥4mm)实行外科手术采卵,滤泡穿刺吸卵共获147个卵细胞,平均每头猴采卵11.31±6.47个,卵细胞回收率为85.96％。精液用直肠电刺激法采集,精子经TALP培养基洗涤二次,并在含10％灭活猴血清的TALP液中,加入1mM CAMP和lmM咖啡因,预孵育6～3h。卵细胞在含10％灭活猴血清的TALP或Ham'sF-10培养基中作体外成熟的培养,经约4～12h孵育,卵细胞体外成热率为28.57％。成熟的卵细胞(n=42,中期Ⅱ)和已预孵育的精子一起12～24h,并有部分在前次输精后5～12h再次输精,在42～56h后有9个受精卵继续发育至2细胞期和桑椹期阶段,体外受精率为35.71％(15/42)。     

人巨细胞病毒对单纯疱疹病毒1型抗原表达的影响

我们用免疫胶体金色埋前标记技术和免疫荧光技术研究了人胚肺细胞(HEL)内,人巨细胞病毒(HCMV-AD_(169))对单纯疱疹病毒1型(HSV-ⅠSM_(44))抗原表达的影响,旨在探讨在细胞这一微生境内,一病毒对另一病毒可能发生的影响。电镜下计数HSV-1组和HCMV HSV-1组特异性结合金颗粒数得HSV-1组为657个,HCMV HSV-1组的总数为283个。t检验P<0.01,差别非常显著。并且HSV-1组细胞的胞浆中的病毒颗粒,比HCMV HSV-1组明显多。荧光显微镜下:HSV-1组阳性细胞数为689个HCMV HSV-1组只有484个,经poisson分布u检验,P<0.01,差别非常显著。免疫荧光实验还表明:HSV-1组,抗血清在1:320时仍有荧光清晰的阳性细胞,而HCMV HSV-1组,抗血清在1:160时,却无荧光阳性细胞。细胞病变效应(CPE)动态观察显示:HSV-1组8小时即有细胞病变,24小时蔓延整个单层;而HCMV HSV-1组超感染14小时才有细胞病变。24小时约有75％细胞受累。结果表明HCMV对HSV-1的抗原表达有明显的抑制作用。对抑制作用的可能机理及其在分子生态学中的意义,进行了讨论。     

Circular RNA SLTM as a miR-421-competing endogenous RNA to mediate HMGB2 expression stimulates apoptosis and inflammation in arthritic chondrocytes

Osteoarthritis (OA) is the most common age-related joint disease characterized by chronic inflammation, progressive articular cartilage destruction, and subchondral sclerosis. Accumulating evidence suggests that circular RNAs (circRNAs) play key roles in OA, but the function of circSLTM in OA remains greatly unknown. Therefore, this study focused on interleukin-1β (IL-1β)-treated primary human chondrocytes as well as a rat model to investigate the expression pattern and functional role of circSLTM in OA in vitro and in vivo. CircSLTM and high mobility group protein B2 (HMGB2) were upregulated in IL-1β-induced chondrocytes, whereas miR-421 was downregulated. Knockdown of circSLTM or overexpression of miR-421 ameliorated IL-1β-induced chondrocyte apoptosis and inflammation. The regulatory relationship between circSLTM and miR-421, as well as that between miR-421 and HMGB2, was predicted by bioinformatics and then verified by the RNA immunoprecipitation experiment and dual-luciferase reporter gene assay. Furthermore, silencing of circSLTM increased cartilage destruction and decreased cartilage tissue apoptosis rate and inflammation in a rat model of OA. Taken together, our findings demonstrate the fundamental role of circSLTM in OA progression and provide a potential molecular target for OA therapy.     

抗人IgG Fc片段及Fab段单抗的鉴定及应用

本实验采用木瓜酶水解,SPA柱亲合层析等手段得到人IgGFc段及Fab段,以Sigma抗人IgGfFc段和抗人IgG Fab段单抗为标准品,鉴定了细胞库中抗人IgG系列的部分细胞株,得到特异性分泌抗人IgG Fc段和抗人IgG Fab段单抗的细胞各一株。 在上述实验基础上,用抗人IgG Fc及抗人IgG Fab单抗分别制备了Sepharose4B亲合层析柱,提纯了酶解人IgG Fc、Fab片段,经ELISA法鉴定,相互之间无交叉反应。同时用此方法制备了人抗HBe Fab片段,并将该片段进行了过氧化物酶标记,用来配制HBe ELISA诊断盒,证明其生物活性未受影响,而且消除了类风湿因子引起的HBe Ag假阳性现象。因抗HBe单抗来源困难,如采用HBe多抗制备ELISA试剂,本法将是提高质量的一个好方法。     

Dieckol,a natural polyphenolic drug,inhibits the proliferation and migration of colon cancer cells by inhibiting PI3K,AKT, and mTOR phosphorylation

This study investigated that dieckol (DKL), a natural drug, inhibits colon cancer cell proliferation and migration by inhibiting phosphoinositide-3-kinase (PI3K), protein kinase B (AKT), and mammalian target of rapamycin (mTOR) phosphorylation in HCT-116 cells. The cells were treated with DKL in various concentrations (32 and 50 μM) for 24 h and then analyzed for various experiments. MTT (tetrazolium bromide) and crystal violet assay investigated DKL-mediated cytotoxicity. Dichlorodihydrofluorescein diacetate staining was used to assess the reactive oxygen species (ROS) measurement, and apoptotic changes were studied by dual acridine orange and ethidium bromide staining. Protein expression of cell survival, cell cycle, proliferation, and apoptosis protein was evaluated by western blot analysis. Results indicated that DKL produces significant cytotoxicity in HCT-116, and the half-maximal inhibitory concentration was found to be 32 μM for 24-h incubation. Moreover, effective production of ROS and enhanced apoptotic signs were observed upon DKL treatment in HCT-116. DKL induces the expression of phosphorylated PI3K, AKT, and mToR-associated enhanced expression of cyclin-D1, proliferating cell nuclear antigen, cyclin-dependent kinase (CDK)-4, CDK-6, and Bcl-2 in HCT-116. In addition, proapoptotic proteins such as Bax, caspase-9, and caspase-3 were significantly enhanced by DKL treatment in HCT-116. Hence, DKL has been considered a chemotherapeutic drug by impeding the expression of PI3K-, AKT-, and mTOR-mediated inhibition of proliferation and cell cycle-regulating proteins.