Ginsenoside improves papillary thyroid cancer cell malignancies partially through upregulating connexin 31

Connexin 31 (Cx31) is considered a suppressor for many tumors. Ginsenoside (Rg1) is a traditional Chinese herb that is widely acknowledged due to its anti-tumor characteristics. However, limited studies have focused on the role of Rg1 in papillary thyroid cancer (PTC) cells. In the current study, we found that the expression of Cx31 in thyroid cancer tissues and thyroid cancer cell lines was significantly lower than that in normal thyroid epithelial tissues and cell lines. Overexpression of Cx31 reduced thyroid cancer cell proliferation, migration and invasion. Furthermore, we found that Rg1 significantly enhanced the expression of Cx31. Moreover, the proliferation and migration of IHH-4 and BCPAP cells were significantly reduced by Rg1 treatment. In contrast, the silencing of Cx31 enhanced the expression of Ki67 and proliferating cell nuclear antigen (PCNA). Meanwhile, treatment with Rg1 significantly decreased the protein levels of Ki67 and PCNA, but these effects could be abolished by transfection with si-Cx31. In summary, we provide novel evidence that the expression of Cx31 was decreased in thyroid cancer cells, but Rg1 treatment could significantly enhance the expression of Cx31 thereby suppressing thyroid cancer cell proliferation and migration.     

抑郁症大鼠海马-杏仁核pMAP-2表达改变及人参皂苷Rb1的干预作用

目的 研究人参皂苷Rb1对抑郁大鼠海马-杏仁核组织微管相关蛋白-2（MAP-2）磷酸化的影响,探讨人参皂苷Rb1抗抑郁作用的可能机制。方法 选用30只成年Wistar大鼠,随机分为对照组、模型组和治疗组。采用慢性不可预性温和应激结合孤养方法建立大鼠抑郁症模型,治疗组同时给予人参皂苷Rb1。采用Western blot方法检测海马、杏仁核组织MAP-2及pMAP-2表达,Real-time PCR检测mRNA表达。结果 模型组大鼠海马和杏仁核组织MAP-2磷酸化程度和mRNA表达水平明显低于对照组（P < 0.05）,治疗组表达明显升高,但仍低于对照组（P < 0.05）。结论 人参皂苷Rb1抗抑郁作用可能与影响海马、杏仁核MAP-2磷酸化有关     

Simultaneous determination of ginsenosides Rg1, Re and Rb1 in rat plasma by HPLC and its application to pharmacokinetic study of SHENMAI injection

In the present study, we developed a sensitive and efficient high performance liquid chromatography (HPLC) method for the simultaneous determination of three ginsenosides (Rg₁, Re, Rb₁) in rat plasma. Chromatographic separation was performed on a C₁₈ (150 mm×4.6 mm) column utilizing gradient elution profile and a mobile phase consisting of (A) water and (B) acetonitrile. The calibration curve, with a great correlation coefficient greater than 0.998, was linear over the range of 1.0–30.0 μg/mL for ginsenoside Rg₁, 0.5–15.0 μg/mL for ginsenoside Re, and 0.5–200.0 μg/mL for ginsenoside Rb₁. The intra- and inter-day precisions for three ginsenosides (Rg₁, Re, Rb₁) were all less than 6.0%, and average recovery, examined at three concentration levels, ranged from 96.1% to 118.6%. The samples was stable within 24 h at 4 ºC storage, and 30 d at –20 ºC storage with three freeze-thaw-assay cycles. The low limits of quantification (LOQ) were 1.0, 0.5 and 0.5 μg/mL for Rg₁,Re and Rb₁, respectively. Taken together, the newly developed method was successfully applied to study the pharmacokinetics of ginsenoside Rg₁, Re and Rb₁ in rat plasma after intravenous administration of SHENMAI injection (SMI).     

Acetylation of Retinoblastoma Like Protein2 (Rb2/p130) in Tumor Tissues

The activity of Rb proteins is controlled by post-translational modifications, especially through phosphorylation.Acetylation of Rb2/p130 was reported recently in NIH3T3 cells but its physiological relevance in cell cycle controland tumorigenesis is still unknown. Efforts are underway to investigate possible interplay between Rb2/p130phosphorylation and acetylation. Here we hypothesized that Rb2/p130 acetylation, like p53 acetylation, may playa role in development of the tumor phenotype. The proposed hypothesis regarding acetylation of Rb2/p130 intumor VS normal cells was found to be true in our case study of 36 tumor samples. Statistical analysis of resultssuggest strong correlation among Rb2/p130 acetylation and cancer phenotype.     

肺癌细胞株Rb蛋白的表达

目的探讨Rb蛋白的表达状况与肺癌细胞分类的关系及肺癌细胞癌变的机制。方法采用Westernblotting方法,检测人类13种小细胞肺癌、9种非小细胞肺癌细胞株Rb蛋白的表达。结果和结论10种小细胞肺癌细胞株存在Rb蛋白的缺失或变异,而非小细胞肺癌细胞株则无此改变,对具有较宽Rb带纹的3种小细胞肺癌和9种非小细胞肺癌细胞株进行了Rb蛋白分型,发现2种小细胞肺癌和7种非小细胞肺癌细胞株以磷酸化型Rb蛋白为主。     

Protective effects of ginsenoside Rg3 on human osteoarthritic chondrocytes

Objectives

To explore whether Rg3, a major and especially potent ginsenoside, modulates human osteoarthritic (OA) chondrocyte senescence.

Methods

Isolated chondrocytes were cultured in medium containing interleukin-1 beta (IL-1β) with or without Rg3. The expression levels of mRNAs encoding aggrecan (ACAN), a major structural proteoglycan, type II collagen (COL2A1), and metalloproteinases (MMP) -1, -3, and -13, respectively, were determined using real-time PCR. Cellular senescence was detected by measuring senescence-associated β-galactosidase (SA-β-Gal) activity. Chondrocyte telomerase activity also served as a senescence marker.

Results

Chondrocytes stimulated by IL-1β showed increased MMP-1, MMP-3, and MMP-13 levels, whereas the expression of COL2A1 and ACAN decreased. However, in cells co-treated with IL-1β and Rg3, the levels of MMP-1 and MMP-13 were lower than in cells treated with IL-1β alone, and COL2A1 and ACAN expression levels recovered from the low values seen when cultured only in the presence of IL-1β. Also, compared to vehicle-treated controls, IL-1β stimulation alone resulted in an increased number of SA-β-Gal-positive cells, while co-incubation with IL-1β and Rg3 significantly suppressed the expression of this senescence marker. Chondrocytes cultured with Rg3 showed significantly higher proliferative and telomerase activities than did control cells.

Conclusions

These findings indicate that Rg3 protects the cell against the development of chondrocyte senescence in osteoarthritis.     

光谱加贝复济治疗化疗引起的难治性口腔溃疡效果观察

目的：评价光谱加复济治疗化疗引起的难治性口腔溃疡的疗效。方法：42例化疗后口腔溃疡患曾接受光谱治疗，效果不佳，再次化疗后又出现口腔溃疡，将其随机分成两组，治疗组接受光谱加贝复济治疗，对照组给予光谱治疗。结果；治疗组总有效率90．48％（19／21），对照组有效率47．62（10／21），两组有效率经秩和检验，差异有显性（P＜0．05），结论：光谱治疗加贝复济对化疗引起的难治性口腔溃疡疗效好，无不良反应，值得推广。     

Differential expression of cell cycle regulators in HCV-infection and related hepatocellular carcinoma

AIM: To investigate cell cycle proteins in chronic hepatitis C virus infection in order to analyze their role in the process of hepatocyte transformation and to characterize their prognostic properties.METHODS: Subjects of the current study included 50 cases of chronic hepatitis C (CHC) without cirrhosis, 30 cases of CHC with liver cirrhosis (LC), and 30 cases of hepatitis C-related hepatocellular carcinoma (HCC) admitted to the Department of Hepato-Gastroenterology, Theodor Bilharz Research Institute (TBRI), Giza, Egypt. Fifteen wedge liver biopsies, taken during laparoscopic cholecystectomy, were also included as normal controls. Laboratory investigations including urine and stool analysis, liver function tests and prothrombin concentration; serologic markers for viral hepatitis and ultrasonography were done for all cases of the study together with immunohistochemical analysis using primary antibodies against Cyclin D1, Cyclin E, p21, p27 and Rb/p105 proteins.RESULTS: Normal wedge liver biopsies didn’t express Cyclin E or Rb/p105 immunostaining but show positive staining for Cyclin D1, p21 and p27. Cyclin D1 expressed nuclear staining that was sequentially increased from CHC to LC (P < 0.01) to HCC (P < 0.001) cases; meanwhile, Cyclin E revealed nuclear positivity only in the case of HCCs patients that was directly correlated to Rb/p105 immuno-reactivity. The expression of p21 and p27 was significantly increased in CHC and LC cases compared to normal controls and HCCs with no significant difference between well- and poorly-differentiated tumors. p21 showed only a nuclear pattern of staining, while, p27 presented with either cytoplasmic and/or nuclear reactivity in all studied cases. Correlation analysis revealed a direct relation between Cyclin D1 and p21 in CHC cases (P < 0.001), between Cyclin D1 and Cyclin E in HCCs (P < 0.01); however, an inverse relationship was detected between Cyclin D1 and p21 or p27 (P < 0.001) and between p21 and Rb/p105 (P < 0.05) in HCCs.CONCLUSION: Upregulation of Cyclin D1 in CHC plays a vital role in the development and differentiation of HCC; while, Cyclin E may be a useful marker formonitoring tumor behavior. p21 and p27 can be used as predictive markers for HCC. Furthermore, higher expression of Rb/p105 as well as inverse relation with p21 and histologic grades suggests its important role in hepatic carcinogenesis.