Preparation of chicken IgY against recombinant E2 protein of bovine viral diarrhea virus (BVDV) and development of ELISA and ICA for BVDV detection

Bovine viral diarrhea virus (BVDV) infects cattle and may lead to persistent infection (PI). The PI animals harbor BVDV throughout their life and become immune tolerant against BVDV. Thus, diagnosis of this virus in herd is highly important. Recombinant E2 protein expression (using pET-32a in Escherichia coli) was confirmed by SDS-PAGE and Western blotting; then purified by Ni⁺ affinity chromatography. Chickens were immunized with BVDV-E2 protein, and IgY antibodies were extracted from egg yolk by PEG-6000. The peak titer of anti-BVDV-E2-IgY was 1:128,000 after the fifth immunization. IgY-based enzyme-linked immuno sorbent assay (ELISA) and immunochromatographic assay (ICA) were further developed. Coincidence of ELISA and ICA test with RT-PCR was 95.45 and 90.91%, respectively. The anti-BVDV-E2 IgY could be used in routine screening of BVDV infection. Besides, it can also be applicable while licensing and/or using live vaccines; screening of imported products containing bovine serum and strong surveillance of BVDV outbreaks.     

江西金盆山自然保护区种子植物区系研究

江西金盆山自然保护区位于江西省龙南县、全南县与信丰县的交汇处,为武夷山脉与南岭山脉之间的过渡区域,保护区植物种类丰富、区系地理成分复杂。经野外实地调查、查阅相关文献资料,对保护区内种子植物的科、属、种分布区类型进行统计分析。结果表明：（1）金盆山保护区共有野生种子植物161科659属1 474种,其中裸子植物6科7属7种,占江西裸子植物总种数（83种）的8.43%,被子植物155科652属1 467种,占江西被子植物总种数（4 369种）的33.58%。（2）区系分析显示,金盆山保护区内科的分布区类型中热带分布和温带分布分别占总科数（除去世界分布）的64.22%和35.77%,热带分布占优势地位;属的分布区类型中热带分布和温带分布分别占总属数（除去世界分布）的50.95%和47.55%,二者接近平衡;种的分布区类型中热带分布种和温带分布分别占总种数（除去世界分布）的33.98%和37.15%,而且中国特有种419种,占总种比的28.88%。（3）金盆山保护区种子植物具有单种属、寡种属多,植物区系起源古老的特点,其中有寡种属212属,占总属比的32.17%;单种属374属,占总属比的56.75%;其中寡种属所含种数537种,占总种比的36.43%。（4）与邻近6个保护区的植物区系比较表明,金盆山保护区与武夷山脉的联系较南岭山脉更近,而且金盆山保护区表现出比邻近的阳际峰等武夷山脉地区热带性质更明显。     

Role of Phosphotyrosine Interaction Domain Containing 1 in Porcine Intramuscular Preadipocyte Proliferation and Differentiation

Phosphotyrosine interaction domain containing 1 (PID1), a recently identified gene involved in obesity-associated insulin resistance, plays an important role in fat deposition. However, its effect on porcine intramuscular preadipocyte proliferation and differentiation remains poorly understood. In this study, the plasmid pcDNA3.1(+)-pPID1 was transfected into porcine intramuscular preadipocytes with Lipofectamine 3000 reagent to over-express porcine PID1 (pPID1). Over-expression of pPID1 significantly promoted porcine intramuscular preadipocyte proliferation. Expression of pPID1 mRNA was significantly increased upon porcine intramuscular preadipocyte differentiation. Indirect fluorescent immunocytochemistry demonstrated that pPID1 protein was localized predominantly in the nucleus of porcine intramuscular preadipocyte. The mRNA levels of peroxisome proliferators-activated receptor γ, CCAAT/enhancer binding protein α and lipoprotein lipase were significantly increased by pPID1 over-expression. Over-expression of pPID1 also led to an increase in lipid accumulation which was detected by Oil Red O staining, and significantly increased the intramuscular triacylglycerol content. These results indicate that pPID1 may play a role in enhancing porcine intramuscular preadipocyte proliferation and differentiation.     

鼻中隔成形术中鼻腔填塞法及鼻中隔“8”字缝合法的疗效比较研究

目的:探讨鼻中隔成形术中鼻腔填塞法及鼻中隔"8"字缝合法的疗效。方法:选取2017年7月~2018年7月期间我院收治的拟行鼻中隔成形术的鼻中隔偏曲患者60例。采用乱数表法将患者分为对照组、实验组两组,各30例患者,两组患者均行鼻内镜下鼻中隔成形术,术后对照组给予鼻腔填塞法处理,实验组给予鼻中隔"8"字缝合法处理。比较两组术后临床疗效、鼻粘膜水肿情况,观察两组不同时间点视觉疼痛模拟评分(VAS)情况,记录两组患者术后鼻中隔并发症发生情况。结果:实验组术后7d临床总有效率为96.67%(29/30),显著高于对照组患者的80.00%(24/30),差异有统计学意义(P0.05)。与术前相比,两组患者术后12hVAS评分升高,术后24h、术后48hVAS评分下降(P0.05);与术后12h相比,两组患者术后24h、术后48hVAS评分均下降,且术后48hVAS评分低于术后24h(P0.05);实验组术后12h、术后24h、术后48hVAS评分均低于对照组(P0.05)。实验组鼻粘膜水肿0级患者数量显著高于对照组,鼻粘膜水肿1级患者数量显著低于对照组(P0.05);两组鼻粘膜水肿2级患者数量比较差异无统计学意义(P0.05);两组患者鼻中隔并发症总发生率比较差异无统计学意义(P0.05)。结论:相较于鼻腔填塞法,鼻中隔成形术中采用鼻中隔"8"字缝合法疗效显著,可减轻患者疼痛,改善患者鼻粘膜水肿情况,且不会增加并发症发生率,具有一定的临床应用价值。     

Strategy for efficient cloning of biosynthetic gene clusters from fungi

Filamentous fungi are excellent sources for the production of a group of bioactive small molecules which are often called secondary metabolites(SMs). The advanced genome sequencing technology combined with bioinformatics analysis reveals a large number of unexplored biosynthetic gene clusters(BGCs) in the fungal genomes. To unlock this fungal SM treasure, many approaches including heterologous expression are being developed and efficient cloning of the BGCs is a crucial step to do this.Here, we present an efficient strategy for the direct cloning of fungal BGCs. This strategy consisted of Splicing by Overlapping Extension(SOE)-PCR and yeast assembly in vivo. By testing 14 BGCs DNA fragments ranging from 7 kb to 52 kb, the average positive rate was over 80%. The maximal insertion size for fungal BGC assembly was 52 kb. Those constructs could be used conveniently for the heterologous expression leading to the discovery of novel natural products. Thus, our results provide an efficient and quick method for the low cost direct cloning of fungal BGCs.     

Melatonin protects vertebral endplate chondrocytes against apoptosis and calcification via the Sirt1‐autophagy pathway

Melatonin is reportedly associated with intervertebral disc degeneration (IDD). Endplate cartilage is vitally important to intervertebral discs in physiological and pathological conditions. However, the effects and mechanism of melatonin on endplate chondrocytes (EPCs) are still unclear. Herein, we studied the effects of melatonin on EPC apoptosis and calcification and elucidated the underlying mechanism. Our study revealed that melatonin treatment decreases the incidence of apoptosis and inhibits EPC calcification in a dose‐dependent manner. We also found that melatonin upregulates Sirt1 expression and activity and promotes autophagy in EPCs. Autophagy inhibition by 3‐methyladenine reversed the protective effect of melatonin on apoptosis and calcification, while the Sirt1 inhibitor EX‐527 suppressed melatonin‐induced autophagy and the protective effects of melatonin against apoptosis and calcification, indicating that the beneficial effects of melatonin in EPCs are mediated through the Sirt1‐autophagy pathway. Furthermore, melatonin may ameliorate IDD in vivo in rats. Collectively, this study revealed that melatonin reduces EPC apoptosis and calcification and that the underlying mechanism may be related to Sirt1‐autophagy pathway regulation, which may help us better understand the association between melatonin and IDD.     

High Thermoelectric Performance in p‐type Polycrystalline Cd‐doped SnSe Achieved by a Combination of Cation Vacancies and Localized Lattice Engineering

Herein, a high figure of merit (ZT) of ≈1.7 at 823 K is reported in p‐type polycrystalline Cd‐doped SnSe by combining cation vacancies and localized‐lattice engineering. It is observed that the introduction of Cd atoms in SnSe lattice induce Sn vacancies, which act as p‐type dopants. A combination of facile solvothermal synthesis and fast spark plasma sintering technique boosts the Sn vacancy to a high level of ≈2.9%, which results in an optimum hole concentration of ≈2.6 × 10¹⁹ cm^?3 and an improved power factor of ≈6.9 µW cm^?1 K^?2. Simultaneously, a low thermal conductivity of ≈0.33 W m^?1 K^?1 is achieved by effective phonon scattering at localized crystal imperfections, as observed by detailed structural characterizations. Density functional theory calculations reveal that the role of Cd atoms in the SnSe lattice is to reduce the formation energy of Sn vacancies, which in turn lower the Fermi level down into the valence bands, generating holes. This work explores the fundamental Cd‐doping mechanisms at the nanoscale in a SnSe matrix and demonstrates vacancy and localized‐lattice engineering as an effective approach to boosting thermoelectric performance. The work provides an avenue in achieving high‐performance thermoelectric properties of materials.     

Effects of Proton Pump Inhibitors on the Gastrointestinal Microbiota in Gastroesophageal Reflux Disease

Proton pump inhibitors(PPIs) are commonly used to lessen symptoms in patients with gastroesophageal reflux disease(GERD). However, the effects of PPI therapy on the gastrointestinal microbiota in GERD patients remain unclear. We examined the association between the PPI usage and the microbiota present in gastric mucosal and fecal samples from GERD patients and healthy controls(HCs) using 16 S rRNA gene sequencing. GERD patients taking PPIs were further divided into short-term and long-term PPI user groups. We showed that PPI administration lowered the relative bacterial diversity of the gastric microbiota in GERD patients. Compared to the non-PPIuser and HC groups, higher abundances of Planococcaceae, Oxalobacteraceae, and Sphingomonadaceae were found in the gastric microbiota from the PPI-user group. In addition, the Methylophilus genus was more highly abundant in the long-term PPI user group than in the short-term PPI-user group. Despite the absence of differences in alpha diversity, there were significant differences in the fecal bacterial composition of between GERD patients taking PPIs and those not taking PPIs. There was a higher abundance of Streptococcaceae, Veillonellaceae, Acidaminococcaceae,Micrococcaceae, and Flavobacteriaceae present in the fecal microbiota from the PPI-user group than those from the non-PPI-user and HC groups. Additionally, a significantly higher abundance of Ruminococcus was found in GERD patients on long-term PPI medication than that on shortterm PPI medication. Our study indicates that PPI administration in patients with GERD has a significant effect on the abundance and structure of the gastric mucosal microbiota but only on the composition of the fecal microbiota.