Novel Compounds Identified by Structure-Based Prion Disease Drug Discovery Using In Silico Screening Delay the Progression of an Illness in Prion-Infected Mice

The accumulation of abnormal prion protein (PrP^Sc) produced by the structure conversion of PrP (PrP^C) in the brain induces prion disease. Although the conversion process of the protein is still not fully elucidated, it has been known that the intramolecular chemical bridging in the most fragile pocket of PrP, known as the “hot spot,” stabilizes the structure of PrP^C and inhibits the conversion process. Using our original structure-based drug discovery algorithm, we identified the low molecular weight compounds that predicted binding to the hot spot. NPR-130 and NPR-162 strongly bound to recombinant PrP in vitro, and fragment molecular orbital (FMO) analysis indicated that the high affinity of those candidates to the PrP is largely dependent on nonpolar interactions, such as van der Waals interactions. Those NPRs showed not only significant reduction of the PrP^Sc levels but also remarkable decrease of the number of aggresomes in persistently prion-infected cells. Intriguingly, treatment with those candidate compounds significantly prolonged the survival period of prion-infected mice and suppressed prion disease-specific pathological damage, such as vacuole degeneration, PrP^Sc accumulation, microgliosis, and astrogliosis in the brain, suggesting their possible clinical use. Our results indicate that in silico drug discovery using NUDE/DEGIMA may be widely useful to identify candidate compounds that effectively stabilize the protein.Electronic supplementary materialThe online version of this article (10.1007/s13311-020-00903-9) contains supplementary material, which is available to authorized users.     

基于UPLC-LTQ-Orbitrap高分辨质谱的葛根芩连汤的化学成分分析

目的基于超高效液相色谱-离子阱-静电场轨道阱质谱(UPLC-LTQ-Orbitrap-MS)的分析方法鉴定葛根芩连汤中化学成分。方法采用色谱柱Dikma Endeavorsil C18(100 mm×2.1 mm,1.8μm),流动相为0.1%甲酸水(A)-乙腈(B),体积流量0.3 mL/min,梯度洗脱。质谱采用ESI源,正负离子模式分别采集一级、二级质谱数据。结果通过对照品指认、软件预测分析,结合文献报道,从葛根芩连汤中鉴定出67个化合物,包括黄酮类36个、生物碱类12个、三萜类及三萜皂苷类4个及其他15个。结论利用UPLC-LTQ-Orbitrap-MS系统阐明葛根芩连汤中化学成分,并初步归纳其各类主要化学成分的质谱裂解特点,为葛根芩根芩连汤的质量控制和作用机制研究提供了参考依据。     

Content of HSP70 in rats with hereditary stress-induced arterial hypertension

Sensitivity of normotensive Wistar rats and NISAG rats (with hereditary arterial hypertension) to heat stress is compared at the organism and cell levels. High temperature sensitivity of NISAG rats correlates with a low content of the main heat shock protein HSP70. This relationship can serve as a biochemical marker of predisposition to arterial hypertension. Translated fromByulleten' Eksperimental'noi Biologii i Meditsiny, Vol. 124, No. 8, pp. 171–173, August, 1997     

肿瘤坏死因子β基因多态性在中国汉族SLE病人中的分布特点及与不同人种的比较研究

目的 :探讨中国江苏地区汉族人群TNFβ基因多态性在SLE病人中分布特点 ,并与不同人种进行比较研究。 方法 :收集江苏地区 16 8名无血缘关系健康个体及 6 6例SLE病人的静脉血提取DNA ,应用聚合酶链反应限制性片段长度多态性(PCR RFLP) ,分析TNFβ基因的多态性。 结果 :中国汉族SLE病人与白种SLE病人TNFβ基因频率差异无统计学意义 ;中国汉族健康个体TNFβ 1等位基因频率明显高于白种健康个体 ;SLE病人TNFβ 2基因频率较正常人明显升高 (SLE病人 6 7 4% ,正常人 5 5 1% ,P <0 0 5 ,R =1 6 8) ;中国汉族SLE病人与白种SLE病人TNFβ基因频率差异无统计学意义。 结论 :TNFβ等位基因频率在正常人分布具有种族差异 ,但在SLE病人分布无种族差异。     

一种简单、稳定、可靠的屏氧酶1基因多态性分析法

目的：建立一种简单、稳定、可靠的PON1基因多态性分析法。方法：取外周静脉血100μl，用ROSE法提取基因组DNA，用两对引物：P192F 5′-TAT TGT TGC TGT GGG ACC TGA G-3′,P192R 5′-CAC GCT AAA CCC AAA TAC ATC TC-3′；P55F 5′-GAA GAG TGA TGT ATA GCC CCA G-3′,P55R 5′-TTT AAT CCA GAG CTA ATG AAA GCC-3分别进行PCR扩增，用限制性内切酶AlwI,NlaⅢ对两种PCR产物分别进行酶切，3％琼脂糖凝胶电泳。结果：扩增的两个PCR产物在192、55多态性位点大小分别为99bp、170bp。Alw I酶切后，凝胶电脉得到完全酶切（66bp,33bp)，部分酶切（99bp、66bp、33bp)，未被酶切（99bp)三种类型DNA片段（RR，QR，QQ基因型）；NlaⅢ酶切后，凝胶电脉得到部分酶切（170bp,126bp,44bp)，未被酶切（170bp)两种类型DNA片段（LM，LL基因型）。经双盲重复检测，结果一致。应用此方法对胃癌患者血样标本检测，发现其PON1基因多态性在192位点频率较高。结论：采用一步PCR－RFLP技术可以建立简单、稳定、可靠的PON1基因多态性分析法。     

Differentiation of Helicobacter pylori isolates by polymerase chain reaction—restriction fragment length polymorphism

Objective: To investigate the association between the diversity of urease gene and urease activity of clinical isolates of Helicobacter pylori (H. pylori). Methods: Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) of urease gene and rapid urease activity test were used to study the urease activity of different clinical isolates of H. pylori. Results: H. pylori clinical isolates were divided into 4 types according to their PCR-RFLP results of urease gene and urease activity. Type Ⅰ , possessing strong urease activity (0. 11) and presented 1 fragment of 1. 7 kb by PCR-RFLP, had close relations with gastric ulcer; type Ⅱ , with the weakest urease activity (0. 07) and 2 fragments (1. 3 and 0. 4 kb respectively) , was associated with duodenal bulb ulcer; type Ⅲ , with the strongest urease activity (0. 12) and 2 fragments (0. 4 and 0. 17 kb) with or without 1 fragment (0. 23 or 0. 37 kb) , was responsible for gastritis; type Ⅳ , with weak urease activity (0. 09) and 2 fragments     

Adhesive fragment reattachment after orthodontic extrusion: a case report

Abstract –  In the treatment of crown fractures, adhesive fragment reattachment provides a good alternative to other restorative techniques, offering several advantages. The present paper reports a case in which the treatment of a cervical crown fracture was accomplished by reattaching the tooth fragment with a flowable resin composite. Orthodontic root extrusion was performed with a modified Hawley appliance prior to fragment reattachment. The clinical and radiographic results after 2.5 years were successful.     

STUDIES OF HLA-D REGION DNA POLYMORPHISM IN INSULIN-DEPENDENT DIBETES MELLITUS

Restriction fragment length polymorphisms (RFLPs) of peripheral leucocyte DNA in insulin-dependent diabetes mellitus (IDDM) patients and in controls were studied with Southern blot analysis. IDDM has been reportedly associated positively with the HLA-DR3 and-DR4 in Caucasians, with the HLA-DR3 and-DR9 in Chinese, and negatively with the HLA-DR2 in both Caucasians and Chinese, Using an HLA-DQβcDNA probe, an EcoRI 2.2 kb fragment is found associated with DR2 controls (r = 0.78, P=1×10~(-6)), but not with DR2 IDDM patients, their respective frequencies being significantly different (P=0.02). The authors also report here their new findings that the frequencies of the EcoRI3.0 kb and BamHI3.3 kb fragments are both decreased in the patient group (P=0.0032 and P=0.0018 respectively). This suggested certain kinds of mutation such as partial deletion or others in the IDDM patients might have caused the RFLP pattern change as compared to the normal DNA. RFLPs may provide valuable information in searching for the genetic basis of IDDM.     

RENIN GENE POLYMORPHISM ASSOCIATED WITH ALDOSTERONE RESPONSIVENESS TO THE RENINANGIOTENSIN SYSTEM IN PATIENTS WITH ALDOSTERONE-PRODUCING ADENOMAS

1. Aldosterone levels in patients with unilateral aldosterone-producing adenomas may be responsive or unresponsive to the renin-angiotensin system, with the former often previously misdiagnosed as bilateral adrenal hyperplasia. 2. In tumours from patients in the responsive subgroup, renin mRNA is expressed in greater amounts than in tumours from patients in the unresponsive subgroup, or in normal adrenals. 3. We compared the frequency of four renin gene polymorphisms in peripheral blood DNA from the two subgroups and found significant associations between BglI, TaqI and HinfI restriction fragment length polymorphisms (RFLP) and aldosterone responsiveness. 4. Allelic variation in the constitutive renin gene was associated with a specific cause of hypertension.