To investigate the effects and underlying molecular mechanisms of the interaction between the non-structural protein 1 (NS1) and nucleolar and coiled-body phosphoprotein 1 (NOLC1) on rRNA synthesis through nucleolar telomeric repeat-binding factor 2 (TRF2) under nucleolar stress in avian influenza A virus infection. The analysis of TRF2 ties into the exploration of ribosomal protein L11 (RPL11) and mouse double minute 2 (MDM2) because TRF2 has been found to interact with NOLC1, and the RPL11-MDM2 pathway plays an important role in nucleolar regulation and cellular processes. Both human embryonic kidney 293T cells and human lung adenocarcinoma A549 cells were transfected with the plasmids pCAGGS-HA and pCAGGS-HA-NS1, respectively. In addition, A549 cells were transfected with the plasmids pEGFP-N1, pEGFP-N1-NS1, and pDsRed2-N1-TRF2. The cell cycle was detected by flow cytometry, and coimmunoprecipitation was applied to examine the interactions between different proteins. The effect of NS1 on TRF2 was detected by immunoprecipitation, and the colocalization of NOLC1 and TRF2 or NS1 and TRF2 was visualized by immunofluorescence. Quantitative real-time PCR was conducted to detect the expression of the TRF2 and p21. There is a strong interaction between NOLC1 and TRF2, and the colocalization of NOLC1 and TRF2 in the nucleus. The protein expression of NOLC1 in A549-HA-NS1 cells was lower than that in A549-HA cells, which was accompanied by the upregulated protein expression of p53 in A549-HA-NS1 cells (all p < .05). TRF2 was scattered throughout the nucleus without clear nucleolar aggregation. RPL11 specifically interacted with MDM2 in the NS1 group, and expression of the p21 gene was significantly increased in the HA-NS1 group compared with the HA group (p < .01). NS1 protein can lead to the reduced aggregation of TRF2 in the nucleolus, inhibition of rRNA expression, and cell cycle blockade by interfering with the NOLC1 protein and generating nucleolar stress.

ETHNOPHARMACOLOGICAL RELEVANCE:Pilose antler is a traditional Chinese medicine used to improve kidney function, strengthen tendons and bones, and prolong life, among other uses. It is widely employed in the treatment of osteoporosis. However, the molecular mechanisms underlying the treatment of high turnover osteoporosis are not fully understood.AIM OF THE STUDY:The present study aimed to investigate the molecular mechanism underlying pilose antler polysaccharide and polypeptide extracts in inhibiting bone resorption in high turnover osteoporosis, and compare the effects of the two components alone and in combination to explore whether they could produce synergistic enhancement effects.MATERIALS AND METHODS:The quantitative and qualitative characteristics of pilose antler polysaccharide and polypeptide extracts were detected by UV-visible spectrophotometry and high-performance liquid chromatography. A rat model of retinoic acid-induced osteoporosis was used to evaluate the inhibitory effect of the extracts on bone resorption. Enzyme-linked immunosorbent assay (ELISA) was used to detect the activity of factors related to high turnover type osteoporosis in rat serum. Western blotting was used to detect the expression of proteins related to the MAKP and MMP-9 signaling pathways in rat femurs. Fluorescence quantitative PCR was used to detect the transcription levels of genes related to the MAKP and MMP-9 signaling pathways in rat femur tissues. Hematoxylin and eosin staining were used to observe the osteoprotective effects of pilose antler polysaccharides and polypeptides.RESULTS:The yield of pilose antler polysaccharides was 8.3%, and was mainly composed of mannose, glucosamine hydrochloride, glucuronic acid, Galacturonic acid, Galactose hydrochloride, glucose, and galactose. The yield of the polypeptides was 26.2%, and eighty percent of the molecular weight of the antler polypeptides was 1.6 kDa-7kD, among which, the molecular weight of 7kD peptide accounted for 52% of the total. Both polysaccharides and peptides could reduce the activities of TRACP, OCN, ERK1, JNK, and MMP-9 in rat serum and reduce both the protein expression and gene transcription levels of ERK1, JNK, and MMP-9 in rat femur tissue with significant differences compared with the model group. Both extracts exerted significant protective effects on rat femur tissue. The effect of pilose antler polypeptides alone was better than that of polysaccharides either alone or in combination.CONCLUSIONS:Pilose antler polysaccharides, polypeptides, and their mixtures could inhibit the occurrence of bone resorption of high turnover osteoporosis by stimulating the MAKP and MMP-9 signaling pathways to reduce the expression of the ERK1, JNK, and MMP-9 genes and proteins, and could help alleviate bone loss caused by retinoic acid. Pilose antler polypeptides had a stronger effect on inhibiting bone resorption. The combination of the two components did not show synergistic enhancement effect, and the polysaccharide tended to moderate the inhibitory enhancement effect of the polypeptide.

目的:基于电子鼻技术对不同品质鹿茸饮片气味特征进行分析与表征.方法:采用PEN3 型电子鼻系统,分析 22 批鹿茸样品的气味特征,以对传感器响应值为指标,进行传感器区别贡献率分析(Loadings)、主成分分析(PCA)、线性判别分析(LDA).结果:Loadings分析表明,5 个传感器对鹿茸饮片气味特征具有较好的响应,不同规格鹿茸饮片气味差异贡献率主要体现为氮氧化物类、甲烷等短链烷烃、有机硫化物、醇醚醛酮类、芳香成分、无机硫化物等成分;PCA表明不同品种与规格鹿茸饮片,其气味的差异性比较明显;LDA发现不同品种及规格鹿茸饮片(除梅花鹿白片与粉片外)样品的气味差异均较明显,表明构成气味的物质存在差异性.结论:电子鼻技术可阐明不同品质鹿茸饮片气味的物质基础;不同品质鹿茸饮片气味存在差异性可揭示鹿茸饮片气味的科学内涵并为其质量控制提供参考.

目的:对马鹿茸多肽A的表面结构进行分析,并探讨其结构对药效的作用.方法:采用原子力显微镜观察马鹿茸多肽A的表面结构.结果:马鹿茸多肽A表面呈三维的曲面、板状结构;其基部为不规则网格状结构,3D结构显示其网格边缘具有粗细不均、长短不等的支链形成椎体状突起,呈栅状或片状.质量浓度为 0.50、0.25、0.10 μg·mL-1 的马鹿茸多肽A的粗糙度分别为 0.667 2、0.700 0 和 0.790 0,表面高度分别为 14.510 5、39.458 8、6.139 7 nm.在相同波长下,一维各向同性功率谱密度越大,则二维各向同性功率谱密度越大,样品对于测定的响应越灵敏.结论:马鹿茸多肽A的曲面板状结构可增加与作用靶点的接触面积,其栅状或片状椎体支链结构则更有利于和作用靶点的相互接触或渗入而易发挥药效作用.随着质量浓度的增大,其粗糙度逐渐变小;而表面高度没有随质量浓度的变化呈现出规律性的变化.

目的:利用显微特征参数,分析比较鹿角与鹿角脱盘显微结构差异,为鹿角粉与鹿角脱盘粉鉴别,以及两者质地差异性提供显微结构基础.方法:采用光学显微镜观察鹿角脱盘、鹿角及鹿骨粉末显微特征,并测定和比较三者的骨陷窝长/短径比值和骨陷窝面积占比显微特征数.结果:鹿角脱盘骨陷窝长/短径比值为 2.04～2.38、骨陷窝面积比为 35.04%～45.82%;鹿角的骨陷窝长/短径比值为 1.69～2、骨陷窝面积比为 40.60%～61.77%;鹿骨骨陷窝长/短径比值为 1.54、骨陷窝面积比为 71.8%.结论:鹿角、鹿角脱盘及鹿骨中三者显微鉴定参数均具有明显的差异性;表明骨陷窝长/短径比与骨陷窝面积比,不仅可作为鹿角与鹿角脱盘显微鉴定参数;也阐释了鹿角脱盘角质地比鹿角更为坚硬的组织结构基础,即随着骨陷窝形态由圆变长、骨陷窝面积比降低,其骨化程度增加、质地更加坚硬.

无线数码互动系统作为实验教学新的辅助手段,已逐渐被应用到各个学科的实验教学中,它不仅具备显微镜技术优势,同时也兼具计算机网络技术和显微图像采集分析技术的重要特征,功能强大且灵活.文章探讨了无线数码显微互动实验系统的功能和优点以及在中药鉴定学实验教学中的应用效果,其促进学生创新思维、自主学习能力及分析解决问题能力的提高,实现了中药学专业创新型人才培养模式的实践与创新.

中药可通过多途径、多靶点干预糖尿病肾病发展进程.在糖尿病肾病的整个病程中,其基本病机由初期的阴虚燥热逐渐发展为气阴两虚,后期阴损及阳,导致阴阳两虚,其病位主要在肾,病久损伤肾络,气血瘀滞,"热"、"虚"、"瘀"贯穿了整个疾病发展过程.临床用药常以补益、清热、活血化瘀药为主.这些中药已被证实通过抗氧化应激、调节自噬、抗炎等减轻患者肾脏组织损伤,进而减缓糖尿病肾病进程.

目的 根据不同批次哈蟆油药材中氨基酸的含量测定结果对不同商品规格等级的哈蟆油进行质量评价及等级区分.方法 采用柱前衍生化-HPLC法同时测定不同规格等级哈蟆油中18种氨基酸的含量.应用系统聚类分析和主成分分析对不同商品规格等级哈蟆油进行类别划分;根据主成分分析结果计算不同批次样品的各主成分得分及综合得分.结果 不同规格等级样品氨基酸总量均在69％～ 82％.通过主成分分析综合得分结果可以得出不同规格哈蟆油药材以氨基酸成分进行分析时线油的评分高于块油;不同等级的块油进行比较得出虽然一等块油中有两个得分较高,但总体来说二等块油的综合得分较一等块油高.结论 应用该方法所测定氨基酸的含量可对不同商品规格等级的哈蟆油进行质量评价及等级区分.

目的 为提高原料利用率,优化鹿茸多肽酶解工艺条件.方法 采用响应面法对多肽酶解体系的酶解温度、pH、加酶量、时间等因素进行考察,以鹿茸多肽含量为评价指标优化鹿茸多肽酶解工艺.结果 鹿茸原料得到充分利用,多肽酶解工艺合理、可行.结论 在50 qC、pH8.0、加酶量3.5％的条件下酶解2h为鹿茸多肽的最佳酶解工艺.

目的 基于电子鼻技术,对九香虫生品和炮制品药材"气"特征进行分析与表征,为九香虫生品和炮制品的质量控制提供实验依据.方法 九香虫样品生品13批、炮制品14批,采用PEN3电子鼻系统,分析九香虫样品的"气"特征,并将所得数据分别进行Loadings、PCA及LDA等统计学分析.结果 在所选的10个传感器中,有5个传感器对九香虫生品及炮制品的"气"特征具有较好的响应,分别为有机硫类、芳香成分、甲烷等短链烃类、小分子氮氧化物类、醇醚醛酮类成分;其中对九香虫生品敏感性较强的为6号传感器,即甲烷等短链烃类成分;对九香虫制品敏感性较强的为9号传感器,即芳香类成分.结论 电子鼻技术可用来解析九香虫药材生制品中"气"特征;九香虫生品与制品的共性成分为机硫化合物,可能为其功效的主要物质;九香虫生品"气"的标志性成分为甲烷等短链烃类成分,九香虫制品"气"的标志性成分为芳香类成分;为揭示九香虫生品和制品"气"的科学内涵及其药材质量控制提供了实验依据.

目的 优化鹿茸糖肽颗粒的制备工艺条件.方法 采用响应面法,以颗粒成型率、流动性、溶化率和吸湿率的综合评分值为评价标准,优化混合辅料比例、辅料用量及乙醇浓度3个制备工艺参数,并加以验证.结果 响应面实验回归模型结果表明各因素对鹿茸糖肽颗粒制备工艺的影响顺序为:辅料用量＞乙醇浓度＞辅料比例.结论 颗粒的最佳制备工艺条件为混合辅料(糊精:微晶纤维素)比值为0.36,辅料用量为主药的3倍,乙醇浓度为50％,制得的颗粒临界相对湿度为76.8％.

在中药学专业教学中建设模拟中药房,是加强中药学专业课程实践教学体系的需要,也是中药学专业创新应用型人才培养目标的需要.模拟中药房是以中医药行业岗位需求为导向,并以学生为主体的模拟仿真实训平台,融教、学、践为一体,促进了中药学专业课程教学质量的提高.该文探讨了模拟中药房在中药鉴定学实践教学中的应用效果,指出了模拟中药房有利于调动学生学习的主动性和积极性,并有利于学生实践能力和综合能力的提高,同时也能在中药学专业创新应用型人才培养中起到积极的促进作用.

哈蟆油具有良好的补益作用,是常用的药食两用药材.由于古代本草文献中对哈蟆油的基原动物记载简略,造成了目前哈蟆油药材使用的混乱.为理清溯源,在查阅本草书籍、文献以及现代中药学专著的基础上,对哈蟆油的名称、基原动物、产地分布、采收加工、功效等方面进行考证.结果 表明哈蟆油因具有补益功效而最早被满族人应用,其基原动物的名称均由满文音译而来.通过分析各类本草书籍中对蛙科动物的描述得出哈蟆油基原动物的最早描述出现于清代阿桂编撰的《盛京通志》之中.在对林蛙属部分动物的分类变迁进行总结归纳后将哈蟆油的基原动物初步确定为东北林蛙Rana dybowskii,黑龙江林蛙R.amurensis和桓仁林蛙R.huanrenensis.根据形态大小可以排除桓仁林蛙,根据药材质量的优劣可以排除黑龙江林蛙,故认为哈蟆油的基原动物为东北林蛙,现主要产区为东北地区,最佳捕捉期为10月份.哈蟆油常炖制后食用.该研究可为哈蟆油基原动物的确定、产区分布和资源利用提供有效依据.

Background: Lotions containing a mixture of glycerin, petroleum jelly, and fatty acids have been shown to improve skin moisturization, alleviate visual dryness, and result in a healthier skin barrier over time. A fuller understanding of the molecular level changes which occur with such improvements has been lacking, particularly in the understanding of lipids, but would provide insights to allow the development of more efficacious products.

目的 分析新西兰鹿茸饮片的白片、骨纱片、红粉片、纱片、蜡片、特级粉片等6种规格饮片中游离、水解氨基酸的种类及含量.方法 采用柱前衍生化、氨基酸自动分析仪,测定样品中游离及水解氨基酸的种类及含量.结果 各样品除“蜡片、骨纱片”外,均检测出Asp、Glu、Gly、Ala、Tyr等5种游离氨基酸,其游离氨基酸总量在1％左右;各样品的水解氨基酸中均未检测出Met、Cys、Ile、Trp、Lys等5种氨基酸,各样品水解氨基酸总量在30％～63％,其中必需氨基酸总量在5％～15％.各规格饮片中水解及游离氨基酸的种类与含量均存在一定差异.结论 新西兰鹿茸饮片中的氨基酸类成分,无论游离氨基酸还是水解氨基酸,其种类及含量与国产鹿茸相比均较低,该方法可用于鹿茸饮片中氨基酸类成分的分析.

课程思政是一种教育理念,要求各类课程与思想政治理论课同向同行,形成协同效应.作者通过近些年来的教育和教学研究,对中药学专业核心课程——《中药鉴定学》课程中融入思政教育进行了实践探索,以挖掘教学内容中的"思政元素"以及针对学生的日常行为和学习行为等方面作为课程思政教育的切入点,提供了可供借鉴的教学经验;并提出了个人对于开展课程思政的思考.为中药学专业相关课程中融入思政教育提供了参考.

目的 考察鹿茸不同组分对去卵巢大鼠骨质疏松症的影响,明确鹿茸健骨作用的主要物质基础.方法 雌性SD大鼠56只,随机分为7组,分别为正常组、模型组、仙灵骨葆组、补佳乐组、鹿茸多糖组、鹿茸多肽组、鹿茸多糖多肽组.采用切除大鼠双侧卵巢的方法复制骨质疏松症模型,连续给药12周后,观察鹿茸不同组分对大鼠骨密度、血清生化指标的影响.结果 连续给药12周后,鹿茸不同组分均能显著提高去卵巢大鼠血清中ALP、OT、BMP-2、E2活性,抑制TRACP、PPARγ2活性,提高去卵巢大鼠的骨密度,鹿茸多肽效果优于鹿茸多糖.结论 鹿茸不同组分对去卵巢所致的大鼠骨质疏松症均有拮抗作用.

This study was carried out to evaluate the preventive and curative effects of Pilose antler against osteoporosis due to kidney deficiency, and investigate its potential mechanism of action. A model of osteoporosis due to kidney deficiency was established in rats using bilateral ovariectomy. Pilose antler polypeptide (PAP), Pilose antler polysaccharide (PAP'), and their mixture (PAP+PAP') were separately administered to the rats for 12 weeks, with progynova and xianlingubao tablets (XLGB) as the positive control groups. We determined the bone mineral density (BMD) and uterus Index of the rats. Osteoblastic bone metabolism-related indices in serum and bone tissue were measured with ELISA. Western blotting and RT-PCR were used to investigate the protein and mRNA expressions of Bmp-2, Smad1, Smad5, Runx2 in bone tissue. The morphology of bone tissue was determined using immunohistochemical methods. Compared with control group, PAP, PAP', PAP+PAP' increased BMD and regulated bone metabolism indices in serum and bone tissue. Treatment with Pilose antler up-regulated the mNRA and protein expressions of Bmp-2, Smad1, Smad5 and Runx2. Immunohistochemical staining showed that Bmp-2, Smad1, Smad5 and Runx2 were stained brown, indicating that all of them were positive. There were abnormal changes in the protein expressions of Bmp-2, Smad1, Smad5 and Runx2 in bone tissue, which may be an important mechanism underlying the development of kidney deficiency osteoporosis. Moreover, PAP, PAP' and PAP+PAP' had some preventive effects on osteoporosis, probably via the activation of the Bmp-2/Smad1 and Smad5/Runx2 signaling pathways through induction of high expressions of their mRNAs and proteins.

Peptides from Pilose antler aqueous extract (PAAE) have been shown to stimulate the proliferation and differentiation of bone marrow mesenchymal stem cells (BMSCs). However, the underlying molecular mechanisms are not well understood. Here, PAAE was isolated and purified to explore the molecular mechanisms underlying PAAE's effects on BMSCs as well as its osteoprotective effects in ovariectomized rats. Our results showed that PAAE promoted proliferation and differentiation of BMSCs to become osteoblasts by enhancing ALP activity and increasing extracellular matrix mineralization. The trabecular microarchitecture of ovariectomized rats was also found to be protected by PAAE. Quantitative reverse transcription-polymerase chain reaction (Quantitative RT-PCR) results suggest that PAAE also increased the expression of osteogenic markers including, alkaline phosphatase (ALP), runt-related transcription factor 2 (Runx2), osteocalcin (OCN), bone morphogenetic protein-2 (BMP-2), and collagen I (COL-I). Immunoblotting results indicated that PAAE upregulated the levels of BMP-2 and Runx2 and was associated with Smad1/5 phosphorylation. PAAE A at the concentration of 200 μg·mL−1 showed the strongest effect on proliferation and osteogenic differentiation of BMSCs after 48 h. Using matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF MS), we identified the molecular weight of PAAE A and found that it is less than 3000 Da and showed several significant peaks. In conclusion, PAAE activates the BMP-2/Smad1, 5/Runx2 pathway to induce osteoblastic differentiation and mineralization in BMSCs and can inhibit OVX-induced bone loss. These mechanisms are likely responsible for its therapeutic effect on postmenopausal osteoporosis.

目的:了解林蛙主产区的人工驯养现状及存在的问题,探究解决方法.方法:采用实地考察与文献梳理相结合的方法.结果:目前吉林、辽宁两省的林蛙养殖企业主要采用半人工驯养方式,其中在养殖品种、放养期管理以及产业链的升级上存在一定问题.结论:建议采用DArT多样性芯片技术标记或SSR微卫星分子标记以及膜基因芯片等技术,对人工驯养林蛙的优良品种繁育和遗传系谱进行确认;通过引进电子标识技术,对放养期林蛙种群的生长习性、生境特点以及病害、物种侵袭等实时监控;以“产、研、销”结合的方式,解决人工驯养林蛙品种来源、放养期监控及产业链发展等问题.