Quantitative Analysis of DNA Content of Adenocarcinoma Cells in Effusion

DNA content of cells in effusion of 33 cases of mesothelial and adenocarcinomacells was measured by microspectrophotometer.The results showed that compared withmesothelial cells the DNA content of the suspected malignant cells was obviously increa-sed (P<0.01),the DNA content of both highly and poorly differentiate adenocarcinomacells was obviously increased and statistically significant difference existed between them(P<0.01);besides,aneuploidy could be found.It is objective,accurate and sensitive todetermine the character and differentiation degrees of adenocarcinoma cells in effusion byquantitaive analysis of DNA content with microspectrophotometer.     

The leaf extract of Mallotus japonicus and its major active constituent,rutin,suppressed on melanin production in murine B16F1 melanoma

Objective: To find anti-melanogenesis materials used in whitening cosmetics.Methods: The ethanolic leaf extract of Mallotus japonicus(M. japonicus) having an anti-melanogenesis activity was separated by a sephadex LH-20 chromatography. Each fraction was measured for its tyrosinase inhibitory activity together with its polyphenol content using the Folin–Ciocalteu method. The anti-melanogenesis activity of the active fractions was determined by the melanin content in the murine B16F1 melanoma. The active fractions were put together due to similar constituents, and then separated by high performance liquid chromatography using a C-18 ODS column. The major antimelanogenesis compound was identified using1 H and13C-NMR and liquid chromatography-mass spectrometry.Results: The ethanolic leaf extract of M. japonicus showed an anti-tyrosinase activity with a high polyphenol content, resulting in suppression of melanin production in the B16F1 melanoma. The extract was separated and the active compound was identical as rutin based on the1 H,13C-NMR and liquid chromatography-mass spectrometry analysis data. In addition, the rutin treatment with cells reduced the melanin content in a concentration dependent manner without any cell toxicity. The leaf extract of M. japonicus containing rutin would be useful in whitening cosmetics for protection from UV-light exposure to be limiting the accumulation of melanin in skin.Conclusions: The leaf extract of M. japonicus and/or rutin isolated from the extract as a key whitening agent would be useful as a whitening cosmetic material for protecting against disorder skin due to melanin accumulation.     

Cloning,Eukaryotic Expression of Human ISG20 and Preliminary Study on the Effect of Its Anti-HBV

Human ISG20 gene was cloned and the effect of its anti-HBV was primarily studied. The ISG20 gene was amplified from HeLa cells by RT-PCR and recombinant vector expressing ISG20 was constructed by genetic engineering. The overexpression of ISG20 in HepG2 cells was detected by Western blot and the levels of secretion of HBs antigen and HBe antigen tested by ELISA. The results showed that： （1） Sequence of ISG20 cloned was consistent to that published in Genebank; （2） Recombinant vector expressing ISG20 could be expressed in HepG2 cells by transfection; （3） The overexpression of ISG20 protein could reduce the levels of the secretion of HBs antigen and HBe antigen in transfected HepG2 cells. It was suggested that the overexpression of recombinant ISG20 in culture cells could reduce the synthesis of HBV proteins.     

Inhibitory Effect of Melatonin on the Growth of H22 Hepatocarcinoma Cells by Inducing Apoptosis

Whether melatonin not only inhibits the growth of H22 hepatocarcinoma cells but also induces apoptosis in vitro was assessed. The anti-proliferative effects of melatonin on tumor cells was observed by MTT assay and tumor cells growth curve assay. And the apoptosis of the cells was studied by acridine orange fluorescence assay and flow eytometry. The cell cycle of the tumor cells was also observed by flow eytometry. It was found that melatonin could significantly inhibit the growth of H22 hepatoeareinoma cells. Incubated with melatonin, ehromatin condensation of the tumor cells was observed by fluorescence microscopy. Compared with control, the percentage of apoptotic cells was increased, and the proportion of G0/S increased but that of G0/M decreased. It was suggested that melatonin could directly inhibit the growth of H22 hepatoearcinoma cells by inducing apoptosis and extending the length of cell cycle of the tumor cells.     

烧伤后假上皮瘤肉芽肿样病变内毒素测量及其致病机制

AIM: To investigate the content of endotoxin in Pseudo-epitheliomatous hyperplastic granuloma-like lesions and its pathogenic mechanism. METHODS: Endotoxin concentrations in the tissue were measured with chromogenic limulus amebocyte lysate ( LAL), which was modified by perchloric acid (PCA) pretreatment for samples. To observe the effect of supernate from peripheral blood mononuclear cells by single or repeated endotoxin stimulation, on proliferation of cultured fibroblasts peripheral blood mononuclear cells. RESULTS: Endotoxin was detected in the injured skin but not detected in the normal skin. Supernate from rom peripheral blood mononuclear cells by single endotoxin stimulation has stronger effect on proliferation of fibroblats than repeated stimulation. CONCLUSION: Pseudo-epitheliomatous hyperplastic granuloma-like lesions were resulted from the bacteria in the injured shin, the bacteria produced endotoxin which stimulated the local inflammatory cells to produce cytokines, and the cytokines led to the overgrowth of repairing cells in the skin.     

Effects of Serum Containing Xinlikang (心力康) on Angiotensin Ⅱ Induced Hypertrophy in Cultured Neonatal Rat Cardiomyocytes

Objective: To evaluate the effects of Xinlikang (心力康,XLK) on angiotensinⅡ(AngⅡ) induced hypertrophic cultured neonatal rat's cardiomyocyte (CMC). Methods: Primary cultured neonatal rat's CMCs with the purity certified by immunohistochemical technique, were divided into three groups. Rats in the normal control group were untreated; those in the model group were established into hypertrophic models but underwent no treatment; and those in the XLK group were established to hypertrophic models and treated with XLK containing serum obtained from rats with aorta coarctation after 8 days of feeding with XLK. MTT and phase-contrast microscope were used to evaluate the effect of XLK on cell activity, pulsating rhythm and surface area; Atrial natriuretic peptide (ANP) expression was determined by radioimmunoassay; Protein content was determined by Bradford method; and DNA synthesis was detected by flow cytometric assay. Results: Immunohistochemistry results showed that more than 90% of the cells wereα-sarcometin actin stained positive cells. No significant effect of XLK on normal CMC was found. AngⅡcould significantly induce hypertrophy in CMCs, and XLK could significantly decrease the increased surface area and the accelerated pulsating rate in them. ANP expression was 780±38 Mg/L in the model group, and 430±23μg/L in the control group, and the elevated expression of ANP in model rats was significantly decreased in the XLK group;The DNA content in the G0/G1 and G2/M phases was significantly enhanced and at the same time it was accompanied with increase of total protein content in the model rats after being stimulated by AngⅡfor 24 h, showing that serum-containing XLK could also significantly suppress total protein synthesis (P<0. 05). Conclusion: XLK could improve AngⅡmediated pathological growth of CMCs without influencing the growth of normal CMCs, suggesting that XLK is probably an effective drug for treatment of myocardial hypertrophy and heart failure.     

Effect and mechanism of tacrolimus on melanogenesis on A375 human melanoma cells

Background Topical tacrolimus has been used for vitiligo as a common treatment option for more than ten years while the underlying mechanism is still uncertain.The aim of this study was to investigate the direct effects of tacrolimus on the melanogenesis and migration on human A375 melanoma cells.The expression of c-KIT mRNA and protein of human A375 cells were also investigated.Methods The cultured A375 human melanoma cells were randomly assigned to control and tacrolimus treatment groups （10,102,103and 104 nmol/L）.The cell proliferation was measured with Cell Counting Kit-8 assays.Melanin content was measured with NaOH method.Transwell migration assay was used to measure cell migration.The expression of c-KIT mRNA and protein were measured with real-time fluorescence quantitative polymerase chain reaction and immunohistochemistry respectively.Results The cell proliferation of the 103 and 104 nmol/L tacrolimus groups were significantly lower （0.666±0.062 and 0.496±0.038） as compared with the control （0.841±0.110,P 〈0.05）.The mean melanin content in all groups treated with different concentration of tacrolimus （10,102,103,104 nmol/L） increased compared with the control group （P 〈0.05）.Dosedependent increase in cell migration were seen in all tacrolimus-treated groups （P 〈0.01）.The expression of c-KIT mRNA level in A375 cells exposed to tacrolimus （103and 104 nmol/L） had significantly increased by 3.03-fold and 3.19-fold respectively compared with the control （P 〈0.05）.Conclusions Although tacrolimus had no effects on cell proliferation on A375 human melanoma cells,it could increase the melanin content and cell migration.The expression of c-KIT mRNA and protein increased dose-dependently in tacrolimus-treated groups as compared with the control.Our study demonstrated that tacrolimus could enhance the melanogenesis and cell migration on A375 cells.     

Effects of Hantaan virus on human endothelial cells and their significance in pathogenesis of hemorrhagic fever with renal syndrome.

The effects of Hantaan virus (HTNV) on human endothelial cells (HECs) were investigated both in vivo and in vitro. The 76-118 strain or SR-11 strain of HTNV were inoculated into HECs monolayers respectively, and the virus antigens could be detected on the seventh day of the first passage after inoculation by immunofluorescent technique. The HTNV could also be isolated through cultures of Vero E-6 cells. HTNV particles and inclusion bodies together with various changes in the organelles were observed in the infected cells by transmission electronic microscopy (TEM), and the immunoenzyme positive virus particles were seen by immunoelectronic microscopy. Samples of skin biopsy were individually obtained from 14 cases of hemorrhagic fever with renal syndrome on the second to the fifth day after the onset of the illness. It was found that HTNV antigens were widely distributed in the cytoplasm of endothelial cells of the samples from 5 out of 14 cases by avidin-biotin-peroxidase complex staining, and morphologic changes of the endothelial cells similar to those observed in vitro, were also seen by TEM. The results indicated that HEC is one of the target cells susceptible to HTNV. The virus could invade and propagate in HECs, and could induce damage to the latter.

     

The cellular and molecular mechanism of laminin-glycopeptides on anti-metastasis

Objective To further study the anti-metastasis mechanism of laminin-glycopeptides on carcinoma cell proliferation, apoptosis and the secretion of matrix metalloproteinases. Methods Human hepatocellular carcinoma cells in serum free medium were incubated on laminin-coated substrate with or without laminin-glycopeptides at a final concentration of 50 μg/ml. The total number of surviving cells after incubating for the indicated time was assayed by MTT assay. DNA synthesis of the incubated cells was detected by (3)H-TdR incorporation.Cell cycle was analysed by FACS.The mitotic index of Giemsa stained cells was assessed.Cell apoptosis was detected by both FACS and an acridine orange staining method.Matrix metalloproteinase secretion was analysed by gelatin zymography. Results The total number of surviving cells incubated on laminin in the absence of laminin-glycopeptides was significantly larger than that in the presence of laminin-glycopeptides. Laminin promoted (3)H-TdR incorporation of carcinoma cells, decreased the percentage of cells in G1 phase and increased the percentage of cells in S phase.In contrast, laminin-glycopeptides could inhibit the effect of laminin as shown by (3)H-TdR incorporation and cell cycle analysis. The percentage of cells in G2+M phase and the mitotic index among various groups showed no significant difference.Matrix metalloproteinases secretion from cells treated by laminin-glycopeptides was much less compared to that without the treatment by laminin-glycopeptides. Conclusion Laminin may stimulate cell proliferation, while laminin-glycopeptides could significantly inhibit the effect of laminin by inhibiting DNA synthesis and arresting the carcinoma cell cycle from G1 to S phase.These effects may inhibit not only tumor growth of the primary carcinoma, but also the establishment of metastases at ectopic tissues. Laminin-glycopeptides could also inhibit the secretion of matrix metalloproteinases from carcinoma cells and this may contribute to their decreased invasive and metastatic phenotype. This study further revealed the cellular and molecular mechanism of laminin-glycopeptides on anti-metastasis.     

竹节人参提取物对衰老模型小鼠的作用

目的研究竹节人参提取物对衰老模型小鼠的影响及抗衰老的作用机制.方法以D-半乳糖衰老模型小鼠和自然衰老模型小鼠为实验对象,检测D-半乳糖衰老模型小鼠心、肝组织中脂褐素(LF)及羟脯氨酸(Hyp)的含量、脑组织中单胺氧化酶(MAO)、谷胱甘肽过氧化物酶(GSH-Px)的活性和一氧化氮(NO)的含量,以及自然衰老模型小鼠脑组织凋亡细胞数.结果竹节人参提取物可减少D-半乳糖小鼠肝脏及心肌组织中LF的含量,升高肝脏组织中Hyp的含量,并可提高脑组织中GSH-Px的活性,降低脑组织中NO的含量和MAO的活性;能明显减少自然衰老模型小鼠脑神经细胞凋亡数目.结论竹节人参提取物有显著的延缓衰老作用,其作用机理与抗氧化、抗自由基、抑制脂质过氧化反应有关.     

竹节人参提取物对异丙肾上腺素致大鼠急性心肌缺血损伤的保护作用

目的：研究竹节人参提取物对异丙肾上腺素(ISO)致大鼠急性心肌缺血的保护作用及机制。方法：采用大剂量ISO皮下注射(SC)诱导产生大鼠急性心肌缺血模型(ISO30mg／kg,每日一次,连续2d),实验大鼠随机分为正常组、ISO模型组、阳性对照组、竹节人参提取物(0．15,0．3g/kg)给药组,测定心脏系数和血清中乳酸脱氢酶(LDH)、磷酸肌酸激酶(CK)和丙二醛(MDA)含量,观察心肌组织谷胱甘肽过氧化物酶(GSH-Px)、Na^ -K^ -ATP酶和Ca^2 -Mg^2 -ATP酶活性。结果:竹节人参提取物能显著降低血清中CK和LDH的释放;也能显著降低血清和心脏匀浆中MDA水平,心脏系数显示对心肌水肿有明显的抑制作用：保护心肌GSH-Px;Na^ -K^ -ATP酶和Ca^2 -Mg^2 -ATP酶活性。结论:竹节人参提取物对ISO所致大鼠急性心肌缺血有显著的保护作用。其机制与抗自由基、抑制脂质过氧化反应有关。     

用基因枪法将bar基因导入杜氏盐藻及转基因藻株的检测

目的：探讨基因枪转化方法不同轰击条件和启动子的选择对转化杜氏盐藻的影响。方法：以抗除草剂(bar)基因作为报告基因,用基因枪法将其导入杜氏盐藻。通过草丁膦筛选培养获得转化藻株,对转化藻株进行分析。结果：在氦气压力为100L／min条件下,微弹轰击2次比微弹轰击1次或3次的效果都好;杜氏盐藻碳酸酐酶(CA1)基因启动子可驱动bar基因在杜氏盐藻巾瞬时表达,而双拷贝碳酸酐酶(DCA1)基因启动子可驱动bar基因在杜氏盐藻中稳定表达。结论：在氦气压力为100L／min条件下微弹轰击2次,可能是基因枪法转化杜氏盐藻的较好转化条件。在转基因杜氏盐藻研究中,DCA1基因启动子可能是一种较为理想的启动子类型。     

珠子参地下及地上部分的显微鉴别研究

目的：通过对珠子参地上部分及地下部分的显微鉴别研究，为珠子参及其同属植物的鉴别提供依据。方法：采用显微鉴别方法对其地下及地上部分作了系统的研究，分别对地下部分的节和节间横切面、粉末，地上部分的茎、叶横切面、粉末及叶片的上下表面观等显微特征进行了描述。结果：详细描述了珠子参各部分的显微特征。结论：本研究不仅提高和完善了珠子参地下部分的显微研究，而且为珠子参地上部分的质量标准研究、开发利用、扩大药用资源提供了理论依据。     

不同麦冬假叶树皂甙元的含量比较

麦冬是常用滋阴中药。中国药典(1985年版)载麦冬为百合科沿阶草属植物麦冬Ophiopogon japonicus(L. f. )Ker-Gawl.的块根。据调查,目前国内麦冬主流商品是麦冬O. japonicus和山麦冬Lirlope spicata(Thunb.)Lour.的块根,也有为阔叶山麦冬     

杜氏盐藻核基质的制备

目的：制备盐藻核基质,进而分离核基质结合区(matrix attachment regions)。方法：采用体积分数0．5％ TritonX—100破碎细胞,体积分数15％Percoll分离盐藻细胞核,二碘水杨酸锂(1ithium diiodosalicylate,LIS)抽提核蛋白,SDS—PAGE电泳分析蛋白质成分。结果：分离出了高纯度的盐藻细胞核,电泳分析表明绝大部分核蛋白已被去除。结论：体积分数0．5％TritonX—100能有效破碎盐藻细胞,体积分数15％Percoll可分离高纯度的盐藻细胞核,25mmol／L LIS抽提可除去盐藻绝大部分核蛋白获得核基质。     

CT三维重建改良R.E.N.A.L.评分系统在50例腹腔镜肾部分切除术中的应用

目的 探讨基于CT三维重建技术改良R.E.N.A.L.评分系统在腹腔镜肾部分切除术中的应用价值。 方法 选取山东大学附属山东省立医院泌尿外科2018年9月至2019年11月间进行的T1期肾细胞癌腹腔镜肾部分切除术50例,所有纳入患者术前均行CT三维重建,并分别按照传统R.E.N.A.L.评分系统以及CT三维重建改良R.E.N.A.L评分系统进行术前评分,结果与手术实际难度进行比较,并利用 Spearman秩相关分析改良R.E.N.A.L.评分系统与患者手术时间、住院天数、术中热缺血时间(WIT)、术后血红蛋白下降、术后血肌酐及术后血尿素氮水平之间的相关性。 结果 CT三维重建改良R.E.N.A.L评分与患者的手术时间、WIT以及术后血红蛋白下降程度之间呈正相关(r_s分别为0.570、0.802、0.903,P<0.001),并且改良R.E.N.A.L评分系统的灵敏度、特异度以及约登指数均明显优于传统R.E.N.A.L.评分系统。 结论 CT三维重建改良R.E.N.A.L.评分系统对患者的手术难度的评估较传统R.E.N.A.L.评分系统更加准确,同时对患者围手术期指标有预测作用。     

竹节人参提取物对低氧模型小鼠的影响

目的 观察竹节人参提取物对低氧模型小鼠的保护作用.方法 分别采用异丙肾上腺素、亚硝酸钠、小鼠断头等方法制备低氧模型,观察竹节人参提取物对低氧小鼠耗氧量、存活时间和张口呼吸时间的影响.结果 灌胃给予竹节人参提取物0.3 mg/kg可降低异丙肾上腺素所致心肌低氧小鼠的耗氧量,延长低氧存活时间;增加离体鼠头的张口呼吸次数并延长呼吸维持时间.结论 竹节人参提取物对异丙肾上腺素所致心肌低氧小鼠具有保护作用.     

煮沸处理对毛蚶中甲型肝炎病毒存活的影响

本实验的目的在于了解毛蚶被污染了甲型肝炎病毒后经通常煮沸食用的方法能否全部杀灭病毒。由于病毒主要积聚在贝类的消化系统,本实验人工接种10~5TCID_(50)的甲型肝炎病毒于活毛蚶的胃肠区,经过煮沸5、10、15、20、25、30和45min的处理后,测定毛蚶中甲型肝炎病毒的存活情况。毛蚶中甲型肝炎病毒的提取采用经改良的Johnson方法。病毒检测采用SL_7细胞,培养4至5周后用间接免疫荧光法检测感染细胞中的甲型肝炎病毒抗原。结果表明,煮沸45min后毛蚶中小部分病毒仍能存活,尽管此时接种标本的8管细胞中只有2管阳性,阳性细胞所占百分比仅为3.8％。     

L5178Y和WTK1细胞微核试验在抗诱变测试中的应用

为了研究Ｌ５１７８Ｙ和ＷＴＫ１细胞应用于抗诱变测试系统的可靠性和有效性,按维生素Ｃ（Ｖｃ）和叶绿酸铜钠（Ｃｈｌ）与丝裂霉素Ｃ（ＭＭＣ）的加入顺序,分为同时处理、预处理三种处理方式,观察Ｖｃ和Ｃｈｌ对ＭＭＣ诱导两种细胞微核抑制作用,结果表明：（１）Ｖｃ和Ｃｈｌ本身无诱导两种细胞微核的作用。（２）Ｖｃ与ＭＭＣ同时处理和预处理,能显著降低ＭＭＣ诱导的两种细胞微核率;（３）０．１２ｍｇ／ｍｌ Ｃｈｌ与ＭＭ