人工培育蝉花虫草的抗肿瘤活性

以天然蝉花虫草生药为对照,以中国仓鼠卵巢肿瘤（Chinese hamster ovary,CHO）细胞株为模型,采用刃天青染色法检测人工培育蝉花虫草的抗肿瘤活性。结果表明：人工培育蝉花虫草和天然蝉花虫草生药都具有抗肿瘤活性,且活性成分均为中等极性化合物。对人工培育蝉花虫草中的抗肿瘤活性成分进行分离制备,得到两种化合物,命名为化合物1和化合物2。化合物1经鉴定为虫草素,化合物2经纯度验证为纯品,活性验证结果发现当其质量浓度为50 μg/mL时,对CHO细胞抑制率高达（94.55±2.33）%。     

桂枝茯苓胶囊联合达英-35和枸椽酸氯米芬治疗多囊卵巢综合征合并不孕症的临床观察

目的: 观察桂枝茯苓胶囊联合达英-35和枸椽酸氯米芬(CC)治疗多囊卵巢综合征(PCOS)合并不孕症的临床疗效。方法: 选择2009年2月至2011年2月,在浙江中医药大学附属第二医院妇产科不孕不育门诊就诊的PCOS不孕患者60例,随机分为对照组30例(达英-35和CC)和治疗组30例(桂枝茯苓胶囊联合达英-35和CC),治疗3疗程,促排卵3个周期,观察患者治疗前后的体重指数(BMI)、腰臀比(WHR)、多毛评分、痤疮评分、卵巢体积、血清黄体生成素(LH)、LH/卵泡刺激素(FSH)、总睾酮(TT),性激素结合球蛋白(SHBG)、空腹血糖(FPG)、空腹胰岛素(FINS)的变化,同时观察排卵率和妊娠率。结果: (1)两组患者WHR、多毛评分、痤疮评分统计学上无明显变化(P>0.05),卵巢体积均明显缩小,统计学上有显著意义(P<0.05)。治疗组BMI显著下降(P<0.05),对照组无显著变化(P>0.05)。(2)治疗后两组患者的LH、LH/FSH比值、TT均下降,SHBG均升高,均有统计学意义(P均<0.01),治疗组FINS显著下降(P<0.05),对照组无明显下降(P>0.05)。(3)治疗组的周期排卵率及周期妊娠率均高于对照组,但统计学上均无显著差异(P>0.05)结论:桂枝茯苓胶囊联合达英-35和CC能显著改善PCOS患者的生殖内分泌情况,改善糖脂代谢,增加CC的敏感性,改善排卵率及妊娠率。     

史氏鲟卵巢营养成分分析

目的分析评价鲟鱼卵巢的营养成分和利用价值。方法以采卵后史氏鲟卵巢为研究对象,分别采用常压干燥法、凯氏定氮法、索氏提取法、高温灼烧法测定水分、粗蛋白、粗脂肪、灰分等基本营养成分;采用氨基酸自动分析仪测定氨基酸组成;采用气相色谱串联质谱检测脂肪酸组成;采用电感耦合等离子体质谱法测定常量、微量无机元素。结果史氏鲟卵巢具有高脂肪、低蛋白的特点,这与鱼类肌肉高蛋白、低脂肪的一般特征明显不同;粗蛋白含量为5.17g/100g,必需氨基酸的比例相对较低,但鲜味氨基酸和药效氨基酸比例较高;粗脂肪含量高达29.80 g/100 g,且富含二十碳五烯酸和二十二碳六烯酸等多不饱和脂肪酸;常量无机元素中, K和Na含量较高,微量无机元素中, Zn和Fe含量较高。结论史氏鲟卵巢可用作鱼油、风味制品、微量元素补充型保健品等的生产原料,具有较好的开发利用前景。     

Autoschizis of human ovarian carcinoma cells: scanning electron and light microscopy of a new cell death induced by sodium ascorbate: menadione treatment

Human ovarian carcinoma (MDAH 2774) cells were treated with sodium ascorbate (VC), menadione (VK3), or a combination of both in a ratio 100:1 for 1h and then examined with scanning electron microscopy (SEM) and light microscopy (LM). Light microscopy data corroborated SEM observations, which demonstrated that death of VC+VK3-treated tumor cells occurred primarily by autoschizis. This type of cell death is characterized by a decrease in cell size, cytoplasmic self-excisions, and nuclear and nucleolar morphologic degradations without the formation of apoptotic bodies. Ultimately, cell death results from karyorrhexis and karyolysis. This study illustrates that plasma membrane damage (branching filopodia, blisters, blebs) results from VC treatment; cytoskeletal damage and self-morsellation are caused by VC, VK3 and VC+VK, treatments. The VC treatment results in a 23% decrease in cell diameter while VK3-treated cells decrease cell diameter by 66%. After 1h of VC+VK3 treatment, a heterogenous cell population is found. This population can be resolved into one population whose diameters are 23% smaller than those of sham-treated cells, and a second population whose diameters are approximately twice those of sham-treated cells. This second population is indicative of doublet formation in which the cells appear to be dividing (an early stage of autoschizic cell death). One half of the doublet contains the cell nucleus while the other half consists of cytoplasm and membrane only. The enucleate portion of this doublet will then be excised. When the types of cell death are enumerated following VC+VK3 treatment, 43% of the cells die by autoschizis, 3% by apoptosis, and 1.9% by oncosis. These results confirm that autoschizis is the principal form of cell death that results from the in vitro treatment of human ovarian carcinoma cells with the vitamin combination.     

Occasional transsynaptic viral labeling in the central nervous system from the polycystic ovary induced by estradiol valerate

Increased density of catecholaminergic nerves in the human polycystic ovary has been observed. The aim of the present study was to investigate the distribution of transsynaptically virus-labeled neurons in the central nervous system from the rat polycystic ovary to see whether is it different or not from that of cycling control rats. To induce a polycystic ovary, a single injection of estradiol valerate was given to adult female rats and 30 days later a neurotropic virus was injected into the right ovary. Rats were sacrificed 72 or 96 hours after viral infection. Weight of the ovaries of the estradiol valerate-treated rats was significantly lower compared to controls, and the histology of the ovaries of the treated rats displayed severely atretic large antral follicles. There was almost no viral labeling in the central nervous system from the ovaries showing precystic morphology, in spite of the fact that such altered organs are rich in nerve fibres. It is assumed that presently unidentified factors in the precystic ovary, presumably related to the link between the immune and the nervous system, might be involved in the infectivity of the virus, and thus be responsible for the lack of viral labeling from such an ovary.     

Enhancement of hepatitis B surface antigen production by adenosine 5'-monophosphate in culture of recombinant Chinese hamster ovary cells

Adenosine 5'-monophosphate was found effective in enhancing hepatitis B surface antigen (HBsAg) production in the culture of recombinant Chinese hamster ovary (CHO) cells, but it markedly inhibited cell growth. A culture strategy with periodical medium changes was established to increase HBsAg production and avoid the AMP-induced inhibition of cell growth. The enhanced energy supply might be one of the reasons for the increase in HBsAg production in the presence of AMP.     

3D NIR‐II Molecular Imaging Distinguishes Targeted Organs with High‐Performance NIR‐II Bioconjugates

Greatly reduced scattering in the second near‐infrared (NIR‐II) region (1000–1700 nm) opens up many new exciting avenues of bioimaging research, yet NIR‐II fluorescence imaging is mostly implemented by using nontargeted fluorophores or wide‐field imaging setups, limiting the signal‐to‐background ratio and imaging penetration depth due to poor specific binding and out‐of‐focus signals. A newly developed high‐performance NIR‐II bioconjugate enables targeted imaging of a specific organ in the living body with high quality. Combined with a home‐built NIR‐II confocal set‐up, the enhanced imaging technique allows 900 µm‐deep 3D organ imaging without tissue clearing techniques. Bioconjugation of two hormones to nonoverlapping NIR‐II fluorophores facilitates two‐color imaging of different receptors, demonstrating unprecedented multicolor live molecular imaging across the NIR‐II window. This deep tissue imaging of specific receptors in live animals allows development of noninvasive molecular imaging of multifarious models of normal and neoplastic organs in vivo, beyond the traditional visible to NIR‐I range. The developed NIR‐II fluorescence microscopy will become a powerful imaging technique for deep tissue imaging without any physical sectioning or clearing treatment of the tissue.     

A rapid and simple technique for correlating light microscopy,transmission and scanning electron microscopy of fixed tissues in Epon blocks

A simplified and standardized technique for close correlation between light microscopy (LM), transmission electron microscopy (TEM) and scanning electron microscopy (SEM) is described. Perfusion and immersion fixed tissue specimens were embedded in Epon 812 and cut for conventional LM and TEM. The Epon blocks with remaining tissue were thereafter treated with epoxy solvent (ethanol-NaOH solution) for partial epoxy resin removal only (dissolving rate approx 33μm/h). The blocks with partially blotted tissue specimens were then critically point dried and gold coated for SEM. This method, in an easy way, allows repeated observations with LM, TEM and SEM with preserved fine structure and exact correlation. Since the technique is so simple and there is no need for special equipment the method can easily be adopted in all laboratories with basic SEM standards.     

Functional Activity of Recombinant Forms of Amh and Synergistic Action with Fsh in European Sea Bass Ovary

Although anti-Müllerian hormone (AMH) has classically been correlated with the regression of Müllerian ducts in male mammals, involvement of this growth factor in other reproductive processes only recently come to light. Teleost is the only gnathostomes that lack Müllerian ducts despite having amh orthologous genes. In adult teleost gonads, Amh exerts a role in the early stages of germ cell development in both males and females. Mechanisms involving the interaction of Amh with gonadotropin- and growth factor-induced functions have been proposed, but our overall knowledge regarding Amh function in fish gonads remains modest. In this study, we report on Amh actions in the European sea bass ovary. Amh and type 2 Amh receptor (Amhr2) are present in granulosa and theca cells of both early and late-vitellogenic follicles and cannot be detected in previtellogenic ovaries. Using the Pichia pastoris system a recombinant sea bass Amh has been produced that is endogenously processed to generate a 12–15 kDa bioactive mature protein. Contrary to previous evidence in lower vertebrates, in explants of previtellogenic sea bass ovaries, mature Amh has a synergistic effect on steroidogenesis induced by the follicle-stimulating hormone (Fsh), increasing E2 and cyp19a1a levels.     

Cardiac and Renal SARS-CoV-2 Viral Entry Protein Regulation by Androgens and Diet: Implications for Polycystic Ovary Syndrome and COVID-19

The susceptibility and the severity of coronavirus disease 2019 (COVID-19) caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) are associated with hyperandrogenism, obesity, and preexisting pulmonary, metabolic, renal, and cardiac conditions. Polycystic ovary syndrome (PCOS), the most common endocrine disorder in premenopausal women, is associated with obesity, hyperandrogenism, and cardiometabolic dysregulations. We analyzed cardiac, renal, circulatory, and urinary SARS-CoV-2 viral entry proteins (ACE2, TMPRSS2, TMPRSS4, furin, cathepsin L, and ADAM17) and androgen receptor (AR) expression, in a peripubertal androgen exposure model of PCOS. Peripubertal female mice were treated with dihydrotestosterone (DHT) and low (LFD) or high (HFD) fat diet for 90 days. HFD exacerbated DHT-induced increase in body weight, fat mass, and cardiac and renal hypertrophy. In the heart, DHT upregulated AR protein in both LFD and HFD, ACE2 in HFD, and ADAM17 in LFD. In the kidney, AR protein expression was upregulated by both DHT and HFD. Moreover, ACE2 and ADAM17 were upregulated by DHT in both diets. Renal TMPRSS2, furin, and cathepsin L were upregulated by DHT and differentially modulated by the diet. DHT upregulated urinary ACE2 in both diets, while neither treatment modified serum ACE2. Renal AR mRNA expression positively correlated with Ace2, Tmprss2, furin, cathepsin L, and ADAM17. Our findings suggest that women with PCOS could be a population with a high risk of COVID-19-associated cardiac and renal complications. Furthermore, our study suggests that weight loss by lifestyle modifications (i.e., diet) could potentially mitigate COVID-19-associated deleterious cardiorenal outcomes in women with PCOS.