消乳散结胶囊治疗兔乳腺增生的实验研究

目的:观察消乳散结胶囊(XRSJ)在治疗兔乳腺增生中的作用,探讨其治病机理。方法:将90只日本大耳兔随机分为6组,分别为XRSJ高、中、低剂量组,逍遥丸组,模型和正常对照组,每组15只,前5组用肌内注射己烯雌酚、黄体酮1个月建立兔乳腺增生模型。造模完成后,行XRSJ灌胃给药,分别取XRSJ给药前、XRSJ给药3个月后及XRSJ停药3个月后作为3个时间观测点。观察乳房高度变化;乳腺光、电镜形态学变化;放射免疫法监测血清雌激素水平。结果:与治疗前及模型对照组相比,中、高剂量XRSJ治疗能明显降低乳房高度(P<0.05);降低增生乳腺小叶腺泡数目、导管腺上皮细胞层数、结缔组织和毛细血管数量(P<0.05);减少乳腺上皮细胞胞质内线立体、高尔基体、粗面内质网数量(P<0.05),使部分增生细胞调亡;显著降低血清雌二醇(E2)含量(P<0.01),停药3个月后治疗效果稳定无反弹。结论:XRSJ治疗具有减轻乳腺组织增生病变、降低E2水平、对兔乳腺增生具有显著的治疗作用。     

乳腺增生组织蛋白质组的凝胶内差异显示电泳分析 `

目的：建立具有高分辨率和稳定性的乳腺增生组织蛋白质组的双向电泳图谱,并对其进行差异蛋白质组分析。方法：取乳腺增生病患者增生部位及正常部位乳腺组织,匀浆提取乳腺组织总蛋白,分别用Cy3或Cy5标记,每一对Cy3和Cy5标记样品都与一个Cy2标记的内标等量混合,上样于同一胶中进行电泳分离,经不同光激发下扫描得到不同样品的蛋白质组图谱。所获得的图谱经DeCyder软件进行分析。结果：在乳腺增生病增生的组织中,有12个蛋白质表达水平显著增加,另外3个蛋白质表达水平显著下降。结论：利用DIGE技术可以作胶内时比分析,也可以根据内标消除胶与胶之间的差异,提高统计的可信度;分析所得的15个差异蛋白质可能与乳腺增生疾病的发生与发展有关。     

疏肝健脾法对产后缺乳大鼠催乳素及其受体表达的影响研究

目的:观察疏肝健脾法对产后缺乳肝郁脾虚证大鼠乳腺组织形态学与催乳素(prolactin,PRL)及其受体(prolactin receptor,PRLR)表达的影响。方法:采用甲磺酸溴隐亭溶液+慢性束缚法复制大鼠产后缺乳肝郁脾虚病证结合动物模型,随机分为模型组、空白对照组、西药对照组、中药低、中、高剂量组,每组8只,共6组。药物干预后,采用HE染色及免疫组化法比较不同干预方法对产后缺乳大鼠乳腺组织形态与催乳素及其受体表达的影响,匀浆乳腺组织,采用ELISA法检测乳腺组织催乳素(PRL)、血清催乳素(PRL)、胃泌素(GAS)、血清下丘脑一垂体一肾上腺轴(The hypothalamic-pituitary-adrenal axis,HPA轴)水平变化:促肾上腺皮质激素释放激素(CRH)、促肾上腺皮质激素(ACTH)、皮质醇(COR)。结果:与模型组相比,西药对照组和中药低、中、高剂量组均可促进乳腺组织向泌乳活跃期发育,表现为腺泡腔充盈、腺泡上皮细胞数量增多、结缔组织和脂肪组织退化,同时增加乳腺组织中PRL(空白对照组(145.72±16.332)、中药中剂量组(127.39±10.854)西药对照组(80.79±18.112)、中药低剂量组(89.42±14.183)、中药高剂量组(88.02±12.064)模型组(54.32±7.128))(P0.05)及PRLR(空白对照组(139.76±14.199)、中药中剂量组(129.09±18.755)西药对照组(87.42±8.390)、中药低剂量组(81.54±11.090)、中药高剂量组(85.167±9.494)模型组(60.93±11.767))的表达(P0.05)。与模型组相比,疏肝健脾中药可降低过高的血清CRH、ACTH、COR水平,升高乳腺组织PRL、血清GAS及PRL水平(P0.05)。其中,中药中剂量组疗效优于其他用药各组(P0.05)。结论:疏肝健脾法促进产后缺乳大鼠的泌乳作用机制与增加PRL及其受体表达、调节胃肠功能、抑制HPA轴的亢进状态,从而促进乳腺组织形态向泌乳活跃期发育密切相关。     

蛹虫草菌丝体多糖对乳腺增生患者ER 和PR 表达的影响

目的:探究蛹虫草菌丝体多糖对良性乳腺增生患者增生乳腺组织中雌激素受体(ER)、孕激素受体(PR)表达的影响。方法:选取我院收治的良性乳腺增生患者50例,行随机数字表法随机分为两组,其中对照组行乳癖消临床常规治疗;实验组在乳癖消治疗基础上加用蛹虫草菌丝体多糖联合治疗,检测和比较两组患者治疗前后增生乳腺组织中ER和PR表达的变化情况。结果:治疗后,两组患者增生乳腺组织中ER和PR表达均明显低于对照组,且实验组显著低于对照组,差异均有统计学意义(P0.05)。结论:蛹虫草菌丝体多糖能有效抑制良性乳腺增生症患者增生乳腺组织中ER、PR是表达,为临床治疗良性乳腺增生症提供了新的思路和方法,值得临床应用和推广。     

近代关于垂体黄体化激素分泌调节的研究

好多年来,就已经认识到垂体促性腺激素的分泌受着环境刺激和性腺类固醇的影响。在某些动物种属中,性交可引起排卵、以及光线可改变动情周期就是环境影响的例证。性腺类固醇对促性腺激素分泌的影响是很复杂的,有人报告,随着情况不同,雌激素和黄体酮,可抑制也可刺激促性腺激素的分泌。这方面的进展,曾由于缺乏灵敏的方法而受到阻碍,近来由于 Parlow 发现了一种高度灵敏的黄体化激素(L.H.)的检定方法,卵巢抗坏血酸降低法,大大地便利了我们对控制这种促激素分泌因素的了解。在这篇讨论中,作者拟就本课题的近代知识加以综述。在评论所用的方法之后,就概述影响促黄体化激素分泌的因素,接着讨论此促激素的神经调节,最后,将对促黄体化激素的调节和其他促性腺激素,卵泡刺激素(FSH)和黄体刺激素(LTH)的调节之间加以比较。     

MUC1粘蛋白在乳腺肿瘤中的表达及其生物学意义

目的　探讨MIC1粘蛋白在乳腺肿瘤组织中的表达特征及其生物学和临床意义。方法　应用免疫组织化学ABC法对 15例乳腺非典型增生 ,15例乳腺良性肿瘤 ,44例乳腺癌 (2 1例含癌旁组织 )染色 ,观察MUC1在不同类型乳腺组织中的表达特征及其与雌激素受体和孕激素受体表达的关系。结果　MUC1在所检测的乳腺组织中均表达 ,但在不同病变乳腺组织中其表达的方式和强度不同。在癌旁和非典型增生组织中只表达在近管腔或腺腔侧的胞膜上 ,胞浆未见表达。在乳腺癌组织中整个胞膜和胞浆均表达 ,且表达强度与细胞的恶性化程度成正相关 (P <0 0 1)。浸润性癌中MUC1表达明显强于原位癌 (P <0 0 0 5 ) ;乳腺癌组织中MUC1的表达强度与雌激素受体的表达成正相关 (P <0 0 5 ) ,与孕激素受体表达无关 (P >0 0 5 )。乳腺癌组织中MUC1有旁分泌现象。结论　MUC1表达水平可反映乳腺肿瘤恶性化程度 ,并可能作为乳腺癌治疗效果判断和预后评价的参考指标。     

乳癖散结颗粒联合达那唑治疗乳腺增生老年妇女内分泌激素及体重指数的疗效分析

目的：本研究旨在探讨乳癖散结颗粒联合达那唑治疗乳腺增生老年妇女内分泌激素及体重指数的疗效分析。方法：以2020年4月-2023年4月在我院诊治的102例老年妇女乳腺增生患者,分为对照组(给予达那唑治疗)和研究组(给予乳癖散结颗粒联合达那唑治疗)各51例,对比两组治疗后临床疗效及不良反应,分析内分泌激素、体重指数、免疫功能、疼痛程度及生活质量。结果：(1)治疗后,研究组临床疗效高于对照组(P<0.05);(2)治疗前,两组T、P、E₂、PRL水平比较,差异不显著(P>0.05);治疗后,研究组T、P、E₂、PRL水平优于对照组(P<0.05);(3)治疗后,研究组CD4⁺、IgG、IgA水平优于对照组(P<0.05);(4)治疗后,研究组精神状况、日常生活、食欲、睡眠评分较对照组高(P<0.05);(5)治疗后,研究组不良反应低于对照组(P<0.05)。结论：乳癖散结颗粒联合达那唑能够有效治疗老年妇女乳腺增生患者,可调节内分泌激素水平,不会影响体重指数,值得临床推广应用。     

p16和cyclinD1在乳腺导管非典型增生癌变过程中的表达

目的 探讨乳腺导管非典型增生癌变过程中p16和cyclinD1的变化及其相互关系。方法 应用免疫组织化学方法检溯p16、cyclin成在乳腺各组病变中的蛋白表达情况。结果 乳腺导管单纯性增生、非典型增生组织中p16蛋白表达率明显高于乳腺癌组织,差异有显性。cyclinD1在中,重度非典型增生组表达最为明显,且与乳腺导管单纯性增生组及轻度非典型增生组比较,差异均有显性。p16、cyclinD1蛋白在乳腺各组病变中表达呈负相关关系。结论 p16、cyclinDl在乳腺增生性病变中呈现不同程度表达,其表达强度在一定程度上与细胞的恶性倾向有关,检测其表达水平可作为乳腺导管非典型增生组织恶性转化的一个客观检测指标。其中,cyclinD1蛋白可能是乳腺癌发生过程中的早期分子事件,可作为临床早期发现乳腺癌的免疫学指标。     

VEGF重组质粒pcDNA/V的构建及其在大鼠急性心肌缺血模型中的表达

构建人血管内皮细胞生长因子(VEGF165)真核表达载体,并研究其在细胞水平和大鼠急性心肌梗死动物模型中的表达。利用RT-PCR方法,从人扁桃体组织中扩增人VEGF165基因,构建真核表达载体pcDNA/V。应用脂质体介导的基因转移技术,将pcDNA/V转染至人胚肾细胞(293细胞)中,经G418筛选获得稳定表达的重组质粒细胞克隆。ELISA、Westernblot检测证实重组质粒pcDNA/V能在293细胞中高效表达外源VEGF基因,鸡胚绒毛尿囊膜血管生成实验证实表达产物具有促血管生成的活性。进一步的体内表达研究,建立大鼠急性心肌梗死模型,将重组质粒pcDNA/V、空质粒pcDNA3.1( )分三点注射于梗死交界处心肌内,四周后取材。经免疫组化染色检测,pcDNA/V组在梗死交界区有VEGF阳性表达;电镜观察显示,pcDNA/V组在梗死交界处心肌细胞间有大量毛细血管内皮细胞增生。实验结果表明成功克隆了人VEGF165基因,构建了其真核表达载体。体内、外表达研究证实重组质粒的表达产物具有促血管生成的生物学活性,为VEGF基因治疗缺血性心肌病的研究提供实验基础。     

凋亡相关基因p53、bcl-2在乳腺导管非典型增生中表达的研究

为观察凋亡相关基因-p53,bcl-2在乳腺导管非典型增生及乳腺癌中的表达,探讨其与乳腺癌组织发生的关系。本实验应用原位杂交方法检测凋亡相关基因p53,bcl-2mRNA,应用免疫组织化学方法检测p53蛋白在44例乳腺导管非典型增生组织中的表达,并与6例乳腺导管单纯性增生及26例乳腺癌对比分析,实验结果为,p53mRNA在乳腺导管单纯性增生组织中呈较强表达（66.7%),在乳腺导自欺欺人这非典型增生组织中阳性表达为40%（轻度;55.6%,中度：41.7%,重度;26.1%)。在癌组织中的表达率为19.2%(导管内癌：21.4%,浸润性导管癌：16.7%),p53蛋白在导管单纯性增生组无表达,在导管非典型增生组阳性表达为24%（轻度：11.1%,中度;25%,重度;34.8%)。在癌组织中的阳性表达为38.4%(导管内癌：35.7%,浸润性导管癌：41.7%),bcl-2mRNA在单纯性增生组无表达,在非典型增生组中阳性表达为轻度：11.1%,中度：16.7%,重度：39.1%,在乳腺癌组织中阳性表达为导管内癌;78.6%,浸润性导管癌83.3%。实验结果表明,在乳腺导管重度非典型增生组织中可检测到p53基因有较高的表达缺失,突变及bcl-2mRNA过表达。     

A Unique Protease Cleavage Site Predicted in the Spike Protein of the Novel Pneumonia Coronavirus (2019-nCoV) Potentially Related to Viral Transmissibility

正Dear Editor,In December 2019, a novel human coronavirus caused an epidemic of severe pneumonia(Coronavirus Disease 2019,COVID-19) in Wuhan, Hubei, China(Wu et al. 2020; Zhu et al. 2020). So far, this virus has spread to all areas of China and even to other countries. The epidemic has caused 67,102 confirmed infections with 1526 fatal cases     

Curcuminoids as inhibitors of thioredoxin reductase: A receptor based pharmacophore study with distance mapping of the active site

Curcumin is the yellow pigment of turmeric that interacts irreversibly forming an adduct with thioredoxin reductase (TrxR), an enzyme responsible for redox control of cell and defence against oxidative stress. Docking at both the active sites of TrxR was performed to compare the potency of three naturally occurring curcuminoids, namely curcumin, demethoxy curcumin and bis-demethoxy curcumin. Results show that active sites of TrxR occur at the junction of E and F chains. Volume and area of both cavities is predicted. It has been concluded by distance mapping of the most active conformations that Se atom of catalytic residue SeCYS498, is at a distance of 3.56 from C13 of demethoxy curcumin at the E chain active site, whereas C13 carbon atom forms adduct with Se atom of SeCys 498. We report that at least one methoxy group in curcuminoids is necessary for interation with catalytic residues of thioredoxin. Pharmacophore of both active sites of the TrxR receptor for curcumin and demethoxy curcumin molecules has been drawn and proposed for design and synthesis of most probable potent antiproliferative synthetic drugs.     

Comparative morphology of the carpel in the Liliaceae: Hewardieae, Petrosavieae, and Tricyrteae

The young pistils in the melanthioid tribes, Hewardieae, Petrosavieae and Tricyrteae, are uniformly tricarpellate and syncarpous. They lack raphide idioblasts. All are multiovulate, with bitegmic ovules. The Petrosavieae are marked by the presence of septal glands and incomplete syncarpy. Tepals and stamens adhere to the ovary in the Hewardieae and the Petrosavieae but not in the Tricyrteae. Two vascular bundles occur in the stamens of the Hewartlieae and Tricyrtis latifolia. Ventral bundles in the upper part of the ovary of the Hewardieae are continuous with compound septal bundles and placental bundles in the lower part. Putative ventral bundles occur in the alternate position in the Tricyrteae and putative placental bundles in the opposite. position in the Petrosavieae. The dichtomously branched stigma in each carpel of the Tricyrteae is supplied by a bifurcated dorsal bundle.     

Enterovirus 71 3C proteolytically processes the histone H3 N-terminal tail during infection

Highlights
1. The N-terminal tail of histone H3 is specifically cleaved during EV71 infection.
2. Viral protease 3C is identified as a protease responsible for proteolytically processing the N-terminal H3 tail.
3. Our finding reveals a new epigenetic regulatory mechanism for Enterovirus 71 in virus-host interactions.     

Detection of EBV and HHV6 in the Brain Tissue of Patients with Rasmussen’s Encephalitis

Rasmussen’s encephalitis (RE) is a rare pediatric neurological disorder, and the exact etiology is not clear. Viral infection may be involved in the pathogenesis of RE, but conflicting results have reported. In this study, we evaluated the expression of both Epstein-Barr virus (EBV) and human herpes virus (HHV) 6 antigens in brain sections from 30 patients with RE and 16 control individuals by immunohistochemistry. In the RE group, EBV and HHV6 antigens were detected in 56.7% (17/30) and 50% (15/30) of individuals, respectively. In contrast, no detectable EBV and HHV6 antigen expression was found in brain tissues of the control group. The co-expression of EBV and HHV6 was detected in 20.0% (6/30) of individuals. In particular, a 4-year-old boy had a typical clinical course, including a medical history of viral encephalitis, intractable epilepsy, and hemispheric atrophy. The co-expression of EBV and HHV6 was detected in neurons and astrocytes in the brain tissue, accompanied by a high frequency of CD8⁺ T cells. Our results suggest that EBV and HHV6 infection and the activation of CD8⁺ T cells are involved in the pathogenesis of RE.     

Perinatal Vertical Transmission of Chikungunya Virus in Ruili,a Town on the Border between China and Myanmar

正Dear Editor,Chikungunya virus (CHIKV), an arbovirus in the family of Togaviridae, genus Alphavirus, is transmitted by the A.aegyptii or A. albopictus mosquito, and causes disease in humans characterized by fever, rash, and arthralgia (Silva and Dermody 2017; Suhrbier 2019). It was first reported in 1953 in Tanzania, and caused only a few outbreaks and sporadic cases in Africa and Asia in last century. However, in the epidemic in 2004, CHIKV acquired mutations that conferred enhanced transmission by the A. albopictus mosquito(Schuffenecker et al. 2006). Since then, it has successively caused outbreaks in Africa, the Indian Ocean, South East Asia, the South America, and Europe (Zeller et al. 2016).     

Development of a Novel Reverse Transcription Loop-Mediated Isothermal Amplification Method for Rapid Detection of SARS-CoV-2

In conclusion, the novel visual RT-LAMP assay is a simple, rapid, and sensitive approach for detection of SARS-CoV-2, and it is ready for application in primary care and community hospitals or health care centers, and even patients' own houses in response to the current SARS-CoV-2 epidemic because the assay does not require sophisticated equipment and skilled personnel. Furthermore, it is also ready to be used in fields for screening samples from wild animals and environments to facilitate the identification of potential intermediate hosts that mediate the cross-species transmission of SARS-CoV-2 from bats to humans.