黄芪多糖对C3H10T1/2细胞棕色脂肪分化中长链非编码RNA表达谱的影响

本试验旨在探究黄芪多糖(APS)对小鼠间充质干细胞C3H10T1/2细胞棕色脂肪化过程中长链非编码RNA(lncRNA)表达谱的影响。试验以C3H10T1/2细胞为研究对象,在成脂诱导分化培养液中添加0.4 g/L的APS,以诱导分化1.5 d的细胞构建文库后测序,筛选差异m RNAs和差异lncRNAs,并对差异m RNAs和差异lncRNAs的顺式及共表达作用预测的靶基因进行了功能分析。通过荧光定量PCR随机分析了3个lncRNAs和3个m RNAs的表达水平,验证了测序结果的准确性。研究结果表明,本次测序共得到13 450个lncRNAs和57 776个m RNAs。通过对其表达量、长度、外显子个数和成对重复性检验分析证明了测序数据的可靠性较好。筛选共得到153个差异表达的lncRNAs和1 238个差异表达的m RNAs。结果表明,差异m RNAs主要基因本体(GO)注释富集在53个功能分类中,京都基因与基因组百科全书(KEGG)分析富集在343条通路中。差异lncRNAs顺式及共表达作用预测的靶基因GO注释分别富集在34和33个功能分类中,在分子功能中条目一致。KEGG分析显示,多个基因富集在脂肪代谢和脂肪分化的信号通路中,尤其在胰高血糖素及cAMP信号通路中富集显著。综上表明,APS导致了C3H10T1/2细胞成脂分化中lncRNA表达谱的变化。本研究结果可为进一步解析APS对干细胞的分化调节提供科学依据。     

核磁共振T2-star-mapping成像软骨定量分析联合血清COMP、PⅡANP对膝骨关节炎的诊断价值研究

摘要 目的：探讨核磁共振T2-star-mapping成像软骨定量分析联合血清软骨寡聚基质蛋白（COMP）、ⅡA型前胶原氨基端肽（PⅡANP）对膝骨关节炎（KOA）的诊断价值。方法：选择2021年4月至2022年6月本院收治的KOA患者138例为KOA组,同期在本院体检的健康志愿者126例为对照组。采用西门子1.5T核磁共振扫描仪,行膝关节T2-star-mapping成像,获取股骨、胫骨内外侧髁关节面及髌骨软骨面T2*值,采用酶联免疫吸附法（ELISA）检测血清COMP、PⅡANP水平,比较对照组与KOA组、不同病情严重程度KOA患者T2*值及血清COMP、PⅡANP水平,采用受试者工作特征曲线（ROC曲线）分析T2*值联合血清COMP、PⅡANP对KOA的诊断价值。结果：对照组血清COMP水平、股骨内侧T2*值、股骨外侧T2*值、胫骨内侧T2*值、胫骨外侧T2*值、髌骨软骨面T2*值低于KOA组（P＜0.05）,血清PⅡANP水平高于KOA组（P＜0.05）。重度组血清COMP水平、股骨内侧T2*值、股骨外侧T2*值、胫骨内侧T2*值、胫骨外侧T2*值、髌骨软骨面T2*值高于中度组,且中度组高于轻度组（P＜0.05）;重度组血清PⅡANP水平低于中度组,且中度组低于轻度组（P＜0.05）。血清COMP水平、血清PⅡANP水平、T2*值、联合检测诊断KOA的ROC曲线下面积分别为0.873、0.843、0.898、0.981。结论：KOA患者血清COMP水平、T2*值升高,血清PⅡANP水平降低,其升高或降低程度与病情严重程度有关,联合检测血清COMP、PⅡANP水平及T2*值对KOA早期诊断价值较高。     

基于Sentinel-1和Sentinel-2A的西小山林场平均树高估测

森林资源调查对于我国森林生态系统可持续发展具有重要意义,森林平均高度是森林资源调查的主要结构参数,也是获取难度最大的关键参数之一.为探究联合主被动遥感技术在估测森林平均高度方面的潜力,本研究以吉林省临江市西小山林场为研究区,利用Senti-nel-1 SAR和Sentinel-2A数据,通过提取Sentinel-1的2...     

SV40 T抗原与人U_1和U_2 snRNA基因的特异结合

利用DNA结合免疫分析证明,编码人U_1和U_2 snRNA基因的5′端区域含有与SV40 T抗原特异结合的序列。SV40 T抗原与U_2基因的亲和力大于U_1基因。DNasel足印(footprinting)分析取得与DNA结合免疫分析一致的结果。U_1和U_2基因的5′端区域含有能被T抗原所保护的,免于DNasel降解的序列。这两个基因的两条链上都含有约30bp长的DNA被T抗原所保护。U_1基因被保护的区域在-11bp—-42bp之间,而U_2基因被保护区域是在-58bp—-90bp之间。     

应用ELISPOT方法筛选确定HIV-1B'/C亚型疫苗六种抗原的H-2d限制的T细胞表位

为了筛选和确定用于检测表达HIV-1 B’/C亚型病毒6种抗原(gp160、gag、polr、evt、at和nef)的艾滋病疫苗免疫小鼠后H-2d限制的特异性T细胞表位,本研究使用表达上述6种抗原的复制型DNA疫苗和非复制型重组痘苗病毒疫苗联合免疫BALB/c小鼠,通过矩阵设计将HIV-1 B(C)亚型6种相应抗原全序列肽库分别混合成肽池,使用肽池对免疫小鼠进行IFN-γELISPOT检测,根据检测结果确定肽库中特异反应的优势表位肽。结果显示:筛选到七条针对Gag的特异表位肽,其中有5条与文献报道相同,另2条为新表位肽;筛选到3条针对Pol蛋白特异表位肽,其中一条为新表位肽;筛选到2条针对gp160特异表位肽,其中一条为新表位肽;在Nef肽库中筛选到一条新的表位肽;从Tat肽库中筛选到3条表位肽,这三条肽在肽库中是连续的序列,都包含(或部分包含)网上公布的表位序列;在Rev肽库中没有筛选到能够产生阳性反应的特异性表位肽。本研究使用IFN-γELISPOT方法筛选和确定了可用于检测表达HIV-1 B’/C亚型病毒6种抗原(gp160、gag、pol、revt、at和nef)的艾滋病疫苗免疫小鼠后H-2d限制的特异性T细胞表位。     

2型糖尿病患者血清鸢尾素、摄食抑制因子-1、3-硝基酪氨酸水平与糖脂代谢和阻塞性睡眠呼吸暂停低通气综合征的关系研究

摘要 目的：探讨2型糖尿病（T2DM）患者血清鸢尾素（Irisin）、摄食抑制因子-1（Nesfatin-1）、3-硝基酪氨酸（3-NT）水平与糖脂代谢和阻塞性睡眠呼吸暂停低通气综合征（OSAHS）的关系。方法：选择2020年4月~2021年9月期间中国人民解放军总医院京南医疗区收治的T2DM患者80例作为研究对象,根据多导睡眠图（PSG）检查结果,合并OSAHS的51例患者列为T2DM合并OSAHS组,剩余的29例纳为T2DM未合并OSAHS组。选择同期来中国人民解放军总医院京南医疗区体检的40例健康志愿者作为对照组。对比T2DM患者、对照组的Irisin、Nesfatin-1、3-NT水平,采用Pearson相关性分析显示Irisin、Nesfatin-1、3-NT与糖脂代谢指标的相关性。T2DM患者发生OSAHS的影响因素采用多因素Logistic回归分析。结果：T2DM合并OSAHS组、T2DM未合并OSAHS组Irisin低于对照组,且T2DM合并OSAHS组低于T2DM未合并OSAHS组（P<0.05）。T2DM合并OSAHS组、T2DM未合并OSAHS组Nesfatin-1、3-NT高于对照组,且T2DM合并OSAHS组高于T2DM未合并OSAHS组（P<0.05）。T2DM合并OSAHS组、T2DM未合并OSAHS组糖化血红蛋白（HbAlc）、空腹血糖（FBG）、餐后2 h血糖（2hPG）高于对照组,且T2DM合并OSAHS组高于T2DM未合并OSAHS组（P<0.05）。T2DM合并OSAHS组、T2DM未合并OSAHS组三酰甘油（TG）、低密度脂蛋白胆固醇（LDL-C）、总胆固醇（TC）较对照组高,高密度脂蛋白胆固醇（HDL-C）低于对照组（P<0.05）。Pearson相关性分析结果显示,Irisin与HbAlc、FBG、2hPG呈负相关,Nesfatin-1、3-NT与HbAlc、FBG、2hPG呈正相关（P<0.05）。T2DM合并OSAHS组、T2DM未合并OSAHS组的年龄、合并高血压、体质量指数、AHI、空腹C肽、合并冠心病对比有差异（P<0.05）。Irisin、Nesfatin-1、3-NT、HbAlc、FBG、2hPG、年龄、合并高血压是T2DM患者发生OSAHS的影响因素（P<0.05）。结论：T2DM合并OSAHS患者的Irisin、Nesfatin-1、3-NT水平表达异常,参与着机体的糖脂代谢过程及OSAHS发生,且OSAHS发生同时还受到HbAlc、FBG、2hPG、年龄、合并高血压的影响,可考虑对上述因素进行早期监测,以进行相关干预。     

五灵胶囊联合富马酸丙酚替诺福韦片对慢性乙型肝炎患者氧化应激、CD4+CD25+调节性T细胞和血清MMP-1、MMP-2、TIMP-1的影响

摘要 目的：探讨五灵胶囊联合富马酸丙酚替诺福韦片对慢性乙型肝炎（CHB）患者氧化应激、CD4⁺CD25⁺调节性T细胞和血清基质金属蛋白酶-1（MMP-1）、基质金属蛋白酶-2（MMP-2）、基质金属蛋白酶组织抑制因子-1（TIMP-1）的影响。方法：纳入苏州大学附属传染病医院2021年6月~2022年12月期间收治的CHB患者122例,采用随机数字表法分为对照组（n=61,富马酸丙酚替诺福韦片）和研究组（n=61,五灵胶囊联合富马酸丙酚替诺福韦片）。对比两组疗效、氧化应激指标、CD4⁺CD25⁺调节性T细胞、肝功能指标[总胆红素（TBIL）、谷丙转氨酶（ALT）、谷氨酰转肽酶（GGT）]、血清MMP-1、MMP-2、TIMP-1水平和不良反应发生情况。结果：研究组的临床总有效率高于对照组（P<0.05）。两组治疗后丙二醛（MDA）下降,且研究组低于对照组同时间点;超氧化物歧化酶（SOD）升高,且研究组高于对照组同时间点（P<0.05）。两组治疗后CD4⁺CD25⁺调节性T细胞下降,且研究组低于对照组同时间点（P<0.05）。两组治疗后MMP-1、MMP-2、TIMP-1下降,且研究组低于对照组同时间点（P<0.05）。两组治疗后TBIL、ALT、GGT下降,且研究组低于对照组同时间点（P<0.05）。两组不良反应总发生率组间对比未见差异（P>0.05）。结论：五灵胶囊联合富马酸丙酚替诺福韦片治疗CHB患者,可有效减轻机体氧化应激,调节CD4⁺CD25⁺调节性T细胞和血清MMP-1、MMP-2、TIMP-1水平。     

Noninvasive In Vivo Demonstration of 2-Fluoro-2-Deoxy-d-Glucose Metabolism Beyond the Hexokinase Reaction in Rat Brain by 19F Nuclear Magnetic Resonance Spectroscopy

The metabolism of 2-fluoro-2-deoxy-D-glucose (FDG) in vivo was observed noninvasively in rat brain using 19F nuclear magnetic resonance (NMR) spectroscopy following an intravenous injection of FDG (400 mg/kg). At 3 h after infusion, four resonances with discrete chemical shifts were resolved. Chemical shift analysis of these resonances suggested the chemical identity of two of the resonances to be FDG and/or FDG-6-phosphate and 2-fluoro-2-deoxy-delta-phosphogluconolactone and/or 2-fluoro-2-deoxy-6-phosphogluconate. The chemical identities of the other two resonances remain to be elucidated. The present study indicates that the metabolism of FDG in vivo is more extensive than is previously recognized and demonstrates the feasibility of using 19F NMR spectroscopy to follow the 19F-containing metabolites of FDG in vivo.     

Detection of Mobile Proteins by Proton Nuclear Magnetic Resonance Spectroscopy in the Guinea Pig Brain Ex Vivo and Their Partial Purification

Abstract: Proton nuclear magnetic resonance (¹H NMR) spectroscopy was used to study metabolites of the brain cortex ex vivo. The superfused brain cortex preparation was judged to be metabolically viable on the basis of the ³¹P NMR spectrum (intracellular pH of 7.23 ± 0.03 and phosphocreatine/ ATP ratio of 1.21 ± 0.09). Using'H NMR a group of previously unidentified signals was detectable at 0.94, 1.22, and 1.40 ppm with a water-suppressed spin-echo sequence. These signals had shorter spin-spin relaxation times (51-54 ms) than TV-acetylaspartate and lactate (84-93 ms) and also smaller saturation factors, an indication of shorter spin-lattice relaxation times than the latter two low-molecular-weight metabolites. The unidentified signals also displayed homo-nuclear coupling to other spins in the methine region of the spectrum. Acid extraction of the brain slices or cortex from animals that were killed yielded a mixture of proteins that exhibited NMR properties matching the ¹H NMR signals in the brain cortex. The molecular mass of these thermoresistant, "mobile' proteins, which contained proline plus hydroxy-proline (9-16% of all amino acids), ranged between 8 and 40 kDa. These "new' assigμMents of¹H NMR-detectable compounds may influence interpretation of NMR data of some metabolites, as their signals are in the vicinity of the -CH3 ¹H NMR peaks of lactate and alanine.     

核磁共振波谱法在蛋白质三维结构解析中的应用

蛋白质特定的三维结构与其生物功能密切相关,因此,研究蛋白质的三维结构有助于揭示其生物功能机制。将核磁共振（NMR）波谱法应用于研究溶液状态下蛋白质的三维结构,能够更加准确地揭示蛋白质结构与生物功能之间的关系。本文综述了NMR解析蛋白质三维结构的理论和技术方法,以及NMR结合其他生物物理手段,并辅以分子建模计算法研究蛋白质三维结构的研究进展和最新方法,为精准解析蛋白质的三维结构提供思路及策略。     

High-Resolution Proton Magnetic Resonance Spectroscopy of Rabbit Brain: Regional Metabolite Levels and Postmortem Changes

The changes in 16 cerebral metabolites produced by cardiac arrest and subsequent room temperature autolysis were studied using high-resolution proton nuclear magnetic resonance spectroscopy. Biopsies of rabbit cerebral cortex, cerebral white matter, and cerebellum were quantitatively analyzed for acetate, alanine, gamma-aminobutyric acid, creatine, glutamate, glycine, inositol, lactate, N-acetylaspartate, phosphocreatine, succinate, taurine, and threonine. Of these, N-acetylaspartate and the total creatine pool are the best candidates for use as concentration reference standards linking in vitro to in vivo 1H nuclear magnetic resonance measurements. Both changed little immediately after death, and they varied in a distinctive way among cortex, white matter, and cerebellum.     

Evaluation of a Quantitative Magnetic Resonance Method for Mouse Whole Body Composition Analysis

Objective: To evaluate applicability, precision, and accuracy of a new quantitative magnetic resonance (QMR) analysis for whole body composition of conscious live mice. Research Methods and Procedures: Repeated measures of body composition were made by QMR, DXA, and classic chemical analysis of carcass using live and dead mice with different body compositions. Caloric lean and dense diets were used to produce changes in body composition. In addition, different strains of mice representing widely diverse populations were analyzed. Results: Precision was found to be better for QMR than for DXA. The coefficient of variation for fat ranged from 0.34% to 0.71% compared with 3.06% to 12.60% for DXA. Changes in body composition in response to dietary manipulation were easily detected using QMR. An increase in fat mass of 0.6 gram after 1 week (p < 0.01) was demonstrated in the absence of hyperphagia or a change in mean body weight. Discussion: QMR and DXA detected similar fat content, but the improved precision afforded by QMR compared with DXA and chemical analysis allowed detection of a significant difference in body fat after 7 days of consuming a diet rich in fat even though average body weight did not significantly change. QMR provides a very precise, accurate, fast, and easy‐to‐use method for determining fat and lean tissue of mice without the need for anesthesia. Its ability to detect differences with great precision should be of value when characterizing phenotype and studying regulation of body composition brought about by pharmacological and dietary interventions in energy homeostasis.     

Fingerprinting Analysis of Rhizoma Chuanxiong of Commercial Types using 1H Nuclear Magnetic Resonance Spectroscopy and High Performance Liquid Chromatography Method

The 1H nuclear magnetic resonance (1H NMR) fingerprints of fractionated non-polar extracts (control substance for a plant drug (CSPD) A) from Rhizoma chuanxiong, the rhizomes of Ligusticum chuanxiong Hort., of seven specimens from different sources were measured on Fourier Transform (FT)-NMR spectrometer and assigned by comparing them with the 1H NMR spectra of the isolated pure compounds. The 1H NMR fingerprints showed exclusively characteristic resonance signals of the major special constituents of the plant. Although the differences in the relative intensity of the 1H NMR signals due to a discrepancy in the ratio of the major constituents among these samples could be confirmed by high performance liquid chromatography analysis, the general features of the 1H NMR fingerprint established for an authentic sample of the rhizomes of L. chuanxiong exhibited exclusive data from those special compounds and can be used for authenticating L. Chuanxiong species.     

二维核磁共振谱在多糖结构研究中的应用

二维核磁共振谱（2D NMR）是获取多糖结构信息,尤其是在多糖序列分析方面的有力工具。本文重点介绍了在多糖结构解析中常用的几种2D NMR谱以及2D NMR解析多糖结构的方法。     

Fingerprinting Analysis of Rhizoma Chuanxiong of Commercial Types using ^1 H Nuclear Magnetic Resonance Spectroscopy and High Performance Liquid Chromatography Method

The ^1 H nuclear magnetic resonance （^1 H NMR） fingerprints of fractionated non-polar extracts （control substance for a plant drug （CSPD） A） from Rhizoma chuanxiong, the rhizomes of Ligusticum chuanxiong Hort., of seven specimens from different sources were measured on Fourier Transform （FT）-NMR spectrometer and assigned by comparing them with the ^1 H NMR spectra of the isolated pure compounds. The ^1 H NMR fingerprints showed exclusively characteristic resonance signals of the major special constituents of the plant. Although the differences in the relative intensity of the ^1H NMR signals due to a discrepancy in the ratio of the major constituents among these samples could be confirmed by high performance liquid chromatography analysis, the general features of the ^1H NMR fingerprint established for an authentic sample of the rhizomes of L. chuanxiong e. hibited exclusive data from those special compounds and can be used for authenticating L. Chuanxiong species.     

Proton Nuclear Magnetic Resonance Spectroscopy of Primary Cells Derived from Nervous Tissue

Abstract: Cell culture techniques, high-resolution in vitro ¹H nuclear magnetic resonance (NMR) spectroscopy, and chromatographic analyses were used to compare the properties of purified cell populations derived from the PNS and cortical neurones. Cell cultures were immunocytochemically characterised with specific antibodies to ensure purity of the individual cultures. Spectra of perchloric acid extracts of cultured Schwann cells, perineural fibroblasts, dorsal root ganglion neurones, and cortical neurones displayed several common features. However, statistically significant differences were found by ¹H NMR spectroscopy in most metabolites among the cell types studied. In addition, cells could be distinguished by the presence or absence of certain amino acids. For example, N -acetylaspartate was present in dorsal root ganglion neurones and cortical neurones, γ-aminobutyric acid was present in large amounts in cortical neurones, and Schwann cell spectra displayed a large signal from glycine. These results extend our earlier findings that different cell types of the CNS exhibit highly characteristic metabolite profiles to now include the major cell types of the PNS. These latter cell types also exhibit characteristic metabolite compositions, such that even Schwann cells and oligodendrocyte type 2 astrocyte (O-2A) progenitor cells—precursors of the myelinating cells of the CNS and PNS, respectively—can be readily distinguished from each other.     

Conformations of P2 Protein of Peripheral Nerve Myelin by Nuclear Magnetic Resonance Spectroscopy

High-resolution 1H NMR spectra of P2 protein from bovine peripheral nerve myelin indicate that the protein contains a high degree of tertiary structure in aqueous solution. Denaturation of the protein in urea solutions is a multi-step process. Binding of lysophosphatidylcholine micelles to the protein causes a conformational change and a broadening of NMR peaks from side chains of aromatic amino acid and methionine residues, with much less effect on upfield methyl resonances.