Study on diagnosis criteria of fire-heat syndrome based on receiver operating characteristic curve and principal component analysis.

OBJECTIVE: To establish the diagnostic quantitative criteria for fire-heat syndrome (FHS) of Chinese medicine (CM) based on the receiver operating characteristic (ROC) curve and principal component analysis (PCA). METHODS: The symptoms and signs of FHS cases and healthy subjects from Guangzhou, Henan and Hunan of China were collected through questionnaire, and the diagnostic quantitative score tables were established for the three regions, respectively, with the method of maximum likelihood analysis. The homogeneity test was then performed on the diagnostic score tables for the three regions with ROC curve, and the diagnostic efficiency of diagnostic score tables for the three regions was compared with the prospective test and retrospective test. The method of PCA was adopted to obtain the analysis matrix for classifying the tapes of FHS. RESULTS: Twenty-seven elements of FHS were confirmed through Chi-square test, and the diagnostic score tables for the three regions were established with the method of maximum likelihood analysis on the basis of the collected case data. According to the ROC curve test, the areas under ROC curve of Guangzhou diagnostic score table assessment with candidates in Guangzhou, Henan and Hunan were 0.998, 0.961 and 0.956, respectively. It showed that the diagnostic efficiency of Guangzhou diagnostic score tables was the highest one. With the prospective test, the area under ROC of Guangzhou diagnostic score table was 0.949, and more than any other diagnostic score table. By PCA, FHS was classified into excess fire and deficiency fire, and then classified into syndrome of flaring up of Heart (Xin) fire, syndrome of Lung (Fei)-Stomach (Wei) excess fire, syndrome of deficiency of Liver (Gan)-yin and Kidney (Shen)-yin, and syndrome of deficiency of Lung-yin from the view of viscera. In the retrospective test, the consistency with clinicians' diagnosis was 69.4%, and in the prospective test, it was 70.1%. CONCLUSIONS: The Guangzhou diagnostic score table could be used as the recommended criteria for the diagnosis of FHS. The classification of FHS was basically in conformity with the clinical situation.

[Study on effects of curcumin on expressions of PDGF-BB, PDGFRbeta and ERK1 of HSC].

OBJECTIVE: To observe the effects of curcumin on expression of PDGF-BB, PDGFRbeta and ERK1 of rat Hepatic stellate cell (HSC-T6). METHODS: The cultured HSC were divided into control group, each curcumin treating group. The influence of curcumin on HSC proliferation was detected by MTT assay. The protein levels of PDGF-BB, PDGFRbeta and ERK1 was detected by immunhistochemical examination; The influence of curcumin on expression of PDGF-BB, PDGFRbeta and ERK1 mRNA was detected by RT-PCR. RESULTS: Immunohistiochemical results showed that normal cultured HSC-T6 cell expressed PDGF-BB, PDGFRbeta and ERK1 remarkably. After the intervention of different concentration of curcumin,the masculine degrees of PDGF-BB, PDGFRbeta and ERK1 and the masculine cell population decreased obviously. The RT-PCR results indicated that the expressions of PDGF-BB, PDGFRbeta and ERK1 mRNA could be obviously seen in control and TGF-beta1 stimulus group. After the intervention of different concentration of curcumin, the expression of PDGF-BB, PDGFRbeta and ERK1 mRNA decreased in a dose-dependent manner. CONCLUSION: Curcumin could inhibit the expression of PDGF-BB, PDGFRbeta and ERK1 which might be the mechanism of action curcumin on anti-fibrosis.

[Radioprotective effect of aloe polysaccharides on three non-tumor cell lines].

BACKGROUND & OBJECTIVE: Our previous study showed that aloe polysaccharides (AP) could evidently decrease the mortality of irradiated mice mainly through increasing the amount of hemocytes and ameliorating immune function of mice. Whether AP can protect the cells in vitro from irradiation damage is unknown. This study was to explore radioprotective effect of AP on 3 non-tumor cell lines, and its effect on cell cycle. METHODS: MTT assay was used to detect cytotoxicities of AP to normal human liver cell line Chang Liver (C. Liver), normal human embryo kidney cell line 293, and normal human umbilicus vein endothelial cell line ECV304. The 3 cell lines were treated with AP before or after irradiation. After 7-10 days normal culture, survival rate of cells was calculated by clone formation assay. Cell cycle was analyzed by flow cytometry (FCM) at different time points after irradiation. RESULTS: 293 cells were treated with AP at different time points before and after x-ray irradiation. Survival rate of 293 cells treated with AP 30 min before x-ray irradiation was the highest (64.2%) among all groups. Evident dosage-effect relationship of AP appeared in concentration range of 12.5-50 microg/ml. After treatment of 50 microg/ml of AP, survival rates of 293, ECV304, and C. Liver cells increased from 41.5%, 46.5%, and 40.9% to 49.4%, 72.1%, and 89.1%, respectively. Irradiation caused a distinct G(2)/M block and decreased G(0)/G(1) phase population in 293 and C. Liver cells. In C. Liver cells, pretreatment of 50 mug/ml of AP increased G(0)/G(1) phase population from 31.8% to 43.8%, decreased G(2)/M phase population from 38.5% to 13.8% 6 h after irradiation; and decreased G(2)/M phase population from 22.9% to 8.7% 24 h after irradiation. In 293 cells, the same pretreatment increased G(0)/G(1) phase population from 30.1% to 45.9% 6 h after irradiation, and from 40.4% to 45.2% 24 h after irradiation accompanied by decrease of G(2)/M population from 59.6% to 54.1%. CONCLUSIONS: AP has radioprotective effect on non-tumor cells. This effect might relate to alleviating of cell cycle turbulence.

Aloe polysaccharides mediated radioprotective effect through the inhibition of apoptosis.

Polysaccharides from aloe are always considered an effective radioprotector on irradiation-induced skin damage. The aim of this study was to determine if aloe polysaccharides (AP) have radioprotective effects on normal human cells in vitro and mouse survival in vivo and to explore the mechanism. Pretreatment with 50 microg/ml AP could improve the surviving fraction at 2 Gy (SF2) of three normal cell lines 293, ECV304, and C. liver from 41.5%, 46.5%, and 40.9% to 49.4%, 72.1%, and 89.1%, respectively. AP could also reduce the apoptotic rate of C. liver cells from 9.5% and 43.0% to 2.2% and 10.9% 48 h and 72 h after 2 Gy irradiation, respectively. Western blot analysis showed that pretreatment with AP could block the upregulation of pro-apoptotic p53, Bax, and Bad and the downregulation of Bcl-2 by irradiation. AP could lower thymocyte apoptosis of mice in vivo after 6 Gy irradiation and abrogate the cell cycle perturbation. Fifty mg/kg of AP treatment for 30 min before 7.5 Gy irradiation provided the best radioprotective effect and improved the 30-day survival rate of mice to 86.0%, from 10.0%. AP exerted radioprotective effects in vitro and in vivo through an inhibition of apoptosis.

Protective effects of polydatin against CCl(4)-induced injury to primarily cultured rat hepatocytes.

AIM:To investigate the protective effects of polydatin (PD) against injury to primarily cultured rat hepatocytes induced by CCl(4).METHODS:Rat hepatocytes were separated by methods of liver infusion in vivo and cultured medium (7.5X10(5) cells/mL). Two mL or 0.2mL was added into 24-well or 96-well plates respectively. Twenty-four hours after cell preculture, PD at concentrations of 10(-7) mol/L-10(-4)mol/L was added into each plate. At the same time injury to hepatocytes was induced by adding 10mmol/L CCl(4).Then, 0.1mL or 1mL culture solution was removed from the 96-well or 24-well plates at 6h, 12h, 24h and 48h after CCl14 intoxication respectively for the determination of GPT, GSH and MDA. At 48h, the survivability of rat hepatocytes was assayed by the MTT colormetric method.RESULTS:After CCl(4) challenge, the release of GPT and the formation of MDA in rat hepatocytes markedly increased and maintained at a high level in 48h, whereas PD with different concentrations could markedly inhibit this elevation with 10(-5)mol/L PD having the strongest effects and inhibiting rate was over 50%. PD could also improve the decreased content of GSH caused by CCl(4) in accordance with the doses used. CCl(4) evidently decreased the hepatocyte survivability from 91.0% ± 7.9% to 35.4% ± 3.8%. On the other hand, PD at 10(-7)mol/L-10(-4)mol/L could reverse this change and improve the cell survival rates to 56.1% ± 5.2%, 65.8% ± 5.0%, 88.7% ± 6.8% and 75.2% ± 7.3%, respectively.CONCLUSION: PD at 10(-7)mol/L-10(-4)mol/L could protect primarily cultured rat hepatocytes against CCl(4) induced injury.