Objective To investigate the mechanism of Sut melanocytes growth inhibition in response to xCT -deficiency. Methods TTotal proteins were extracted from xCT-deficient Sut melanocytes and wild melanocytes, respectively, and were separated by 2-dimensional electrophoresis. Altered expression profile of proteins of Sut melanocytes was analyzed by PDQuest software and compared with that of wild melanocytes.Proteins with significant change were chosen to be identified by mass spectrometry and database query.Results Twenty proteins in Sut melanocytes altered significantly compared with wild melanocytes. Ten of the proteins were up-regulated, while the other tens were down-regulated. Four proteins from both up-regulated and down-regulated were identified respectively: up-regulated proteins were Tubulin alpha-1b, S100-A6,Nucleoside, S-formylglutathione, and down-regulated proteins were Calumenin,NDRG1 ,DPYSL2, 14kDa unknown protein. Conclusion The identification of the xCT-deficiency related proteins may provide supporting evidence for the mechanism research of Sut melanocytes' growth inhibition caused by xCT-deficiency.

Objective:To optimize the extraction technology of radix scutellariae. Methods: The extraction of radix scutellariae was scanned by ultraviolet spectrophotometry from 200 to 400nm. The content of baicalin was determined by HPLC. The ultraviolet ab-sorption, baicalin content and extraction rate were used as the indices, and the optimal extraction conditions were investigated by single factor experiments and orthogonal design tests. Results: The optimal extraction conditions were as follows: the ethanol concentration was 60%, the solid-liquid ratio was 1∶40, ultrasound extraction time and temperature was 40 min and 60℃, respectively. Conclusion:The extraction of radix scutellariae has good sunscreen with promising ultraviolet absorption in UVB. Ultrasound extraction has high ex-traction yield with short time, which can be used to extract sun-screening constituents from radix scutellariae.

Objective To investigate the mechanism of endotoxin tolerance (ETT) through observing the expression of OX40 in liver tissues of ETT rats.Methods SD male rats were randomly divided into three groups:normal group (n=6),acute liver failure (ALF) group (n=24) and ETT group (n=24).Lipopolysacharide (LPS) 0.1 mg/kg (ETT groups) or 0.9 ％NaC1 (ALF groups) was administered by five consecutive intraperitoneal injections at 24 h intervals,and at the sixth day,all animals were treated with intraperitoneal injections of D-galactosamine (D-GalN) 800 mg/kg and LPS 8 μg/rat.Blood and liver tissue were collected at 6,12,24 and 48 hours after the injection of D-GalN/LPS.The gene expression of OX40 in the liver was measured by reverse transcription-polymerase chain reaction (RT-PCR).The protein expression of OX40 was estimated by Western blot.The tumor necrosis factor (TNF)-α and interleukin (IL)-6 levels were determined by enzyme-linked immunosorbent assay (ELISA).The data analysis was performed by one-way ANOVA,lease significant difference (LSD) and Dunnett's T3 test.Results The expressions of TNF α and IL-6 were both significantly lower in ETT group compared to ALF group,but still higher than that of control group.The gene expressions of OX40 peaked at 12 hours and decreased gradually in ALF group.The gene expressions of OX40 were significantly lower in ETT group compared to ALF group (6 h:F=5.027,24 h:F=5.539,48 h:F=5.011,all P＜0.05; 12 h:F=36.688; P＜0.01),but still higher than that of normal group.The tendency of OX40 protein expression in ALF group was peaked at 12 hours and decreased at 24 hours.In ETT group,the expression of OX40 was lower,and the difference between ETT group and ALF group had statistical significance (6 h:F=8.658,P＜0.05; 12 h:F=34.611,24 h:F=28.176,48 h:F=16.747; all P＜0.01).Conclusions The level of OX40 is increased in ALF group,while the expressions of OX40,TNF-α and IL-6 are lower in ETT group,which suggested that OX40 may play an important role in the process of ETT.

Objective To study the effect of endotoxin tolerance (ETT) on chemokine receptor 7 (CXCR7) in the liver tissue of rats with acute liver failure (ALF).Methods SD male rats were randomly divided into three groups:normal group,ALF group and ETT group.The rats in the ETT group and ALF group were injected with lipopolysacharide (LPS) 0.1 mg/kg or saline respectively,one time / day for 5 days.At 24 hours after the 5th-day injection,all rats were injected with D-GalN 800 mg/kg and LPS 8μg/rat.Blood sample and liver tissue were collected on 2,6,12,24 and 48 hours after injection.The gene expressions of CXCR7 in the liver were measured by RT-PCR,and the protein expressions of CXCR7 were determined by Western Blot.The data analysis was performed by LSD,Dunnett's t test.Results The histological damage in the liver tissue was significantly mider in ETT group compared to ALF group.The gene expressions of CXCR7 were significantly milder in ETT group compared to ALF group (2 h:F =29.222,6 h:F=166.892,12 h:F=38.975,24h:F=34.603,48 h:F=18.929,allP＜0.01),but still severer than that of normal group.The CXCR7 protein expression in ALF group and ETT group peaked at 24 hours,but the expression of CXCR7 in ETT group was lower compared with that in in ALF group (2h:F=11.155,6 h:F=42.553,12h:F=17.082,all P＜0.01; 24 h:F=7.242,P＜0.05).Conclusions During the process of endotoxin tolerance,LPS pretreatment and D-GalN can decrease the liver injury,down-regulate the expressions of CXCR7mRNA and CXCR7.This suggests that CXCR7 may play an important role in the ETT.

Objective To observe the consistency of T-cell receptor (TCR) genes mutation in lymphocytes in rats after irradiation in vivo and in vitro.Methods A total of 48 female rats were randomly divided into 6 equal groups.Peripheral blood samples from them were collected to separate the lymphocytes and then irradiated to X-ray irradiation with the dose rate of 200 cGy/min at the doses of 0,0.5,0.75,1.0,2.0,and 3.0 Gy,respectively.Then all the lymphocyte samples were cultured for 7 days.Flow cytometry with direct immunofluorescence was used to detect the TCR gene mutation.The levels of TCR gene mutant frequency (TCRMF) of different groups were calculated.Results The TCRMF levels of different groups after irradiation in vivo and in vitro all displayed a dose-dependent manner and there were no significant differences in the TCRMF between different dose irradiation groups(t = -1.1-0.3 ,P ＞0.05).Conclusions A consistency of TCRMF after irradiation in vivo and in vitro is proven.The results of TCRMF of peripheral blood lymphocytes irradiated in vitro by flow cytometry can precisely reflect the TCR genes mutation after whole-body irradiation.

To investigate the changes of serum ghrelin levels and their relationship with body mass index (BMI) ,waist-to-hip ratio(WHR) ,thyroid function, blood glucose, insulin, and insulin resistance parameters in different thyroid functional status. The fasting serum ghrelin, insulin, glucose, and thyroid hormone levels were determined, BMI, WHR, and homeostasis model assessment for insulin resistance index (HOMA-IR) were calculated in 50 hyperthyroid and 30 hypothyroid patients at diagnosis and after normalization of thyroid function. 30 euthyroid subjects served as control. The ghrelin levels in hyperthyroid patients before treatment were lower than that in control group [(63.2±9.6) ng/L vs (80.9±13.9) ng/L,P<0.01]. Multiple regression analysis showed that HOMA-IR was an independent factor related to fasting ghrelin levels (r = -0.314, P = 0. 027). The ghrelin levels were similar in hypothyroid patients and the controls before and after the treatment. It suggests that insulin resistance might be an important factor in regulating serum ghrelin levels in different thyroid functional status.

BACKGROUND:Previous studies have suggested that the risks for coronary atherosclerotic plaque progression and in-stent restenosis are increased in patients with coronary heart disease combined with type 2 diabetes. OBJECTIVE:To explore the predictive factors for in-stent late loss and non-culprit coronary lesion progression in patients with type 2 diabetes mel itus. METHODS:A total of 399 stenting patients were enrol ed, including 179 diabetic patients and 220 non-diabetic patients. The clinical materials, angiography parameters and biochemical markers were col ected. The difference between the two groups was compared, and also we conducted subgroup analysis in the diabetic patients. Low-density lipoprotein cholesterol, hemoglobin A1c, fibrinogen and high-sensitivity C-reactive protein were detected at days 3, 120, 210 and 360 after stenting. RESULTS AND CONCLUSION:Compared with non-diabetic patients, the stent length (P=0.18) was longer and the stent diameter (P=0.002) was smal er in the diabetic patients. The minimal lumen diameters of post-procedure and fol ow-up angiography in the diabetic group were significantly decreased (P=0.001, P=0), and the diabetic patients also showed severe coronary artery stenosis instantly and within the fol ow-up after stenting (P=0.038, P=0.004). The fol ow-up angiography showed that the diabetic patients had more late loss and restenosis (P=0, P=0.097). Furthermore, in the subgroup analysis of diabetic patients, the levels of hemoglobin A1c, fibrinogen and high-sensitivity C-reactive protein were significantly increased in the patients with restenosis and non-culprit lesion progression. These findings indicate that diabetic patients appear to have the higher incidence of restenosis and non-culprit lesion progression. Moreover, hemoglobin A1c, fibrinogen and high-sensitivity C-reactive protein are effective predictors for in-stent late loss and non-culprit coronary lesion progression.

Objective To investigate the changes of serum cardiac troponin I(cTnI)and brain natriuretic peptide(BNP)in heart failure of children with pneumonia and their relationship with heart function.Methods Thirty healthy children aged from 5 months to 3 years old were randomly selected with 17 male and 13 female(healthy group).Thirty children with severe heart failure aged from 3 months to 2 years old were selected at the same time with 21 male and 9 female(heart failure group).Thirty children with ordinary pneumonia aged from 3 months to 3 years old were also sampled with 16 male and 14 female(ordinary pneumonia group).The peripheral bloods of 2-3 mL of all children were taken.The BNP level were measured by enzyme-linked immunosorbent assay and the cTnI level was determined by micro-particle enzyme immunoluminescent.Left ventricular ejection fraction(LVEF) and left ventricular fractional shor-tening(LVFS)were detected by echocardiography.SPSS 11.0 software was used to analyze the data.Results The levels of cTnI [(0.389?0.030) ?g/L] and BNP [(0.572?0.090) ?g/L] of heart failure group increased significantly compared with healthy and ordinary pneumonia group,while their LVEF and LVFS decreased significantly(Pa

OBJECTIVETo investigate the effect of andrographolide derivative Yanhuning (YHN) on Candida albicans biofilms in rats.

METHODThe rat C. albicans biofilms subcutaneous catheter model was established by intraperitoneally injecting YHN (40, 20, 10, 5, 2.5 mg x kg (-1)), with the FLC (80 mg x kg(-1)) positive group as the control group. After 7 d, CFU counting and XTT assay were used to evaluate the effect of YHN on C. albicans biofllms in vivo. Scanning electron microscopy (SEM) was applied to observe the morphological changes in rat biofilms intervened by YHN. The real-time fluorescence quantification PCR was adopted to detect expressions of C. albicans adhesion-related genes, such as ALS1, ALS3, HWP1, EAP1 and MP65.

RESULTThe YHN group showed much less CFUs on catheter pieces and lower XTT metabolic activity than the blank group, with dosage dependence. SEM also showed that YHN could obviously decrease C. albicans adhesion on subcutaneous catheters in rats. According to qRT-PCR's results, YHN can down-regulate expressions of ALS1, ALS3, HWP1, EAP1 and MP65.

CONCLUSIONYHN could inhibit C. albicans biofilms in rats.

Animals ; Biofilms ; drug effects ; growth & development ; Candida albicans ; cytology ; drug effects ; physiology ; Catheters ; microbiology ; Cell Adhesion ; drug effects ; Diterpenes ; chemistry ; pharmacology ; Dose-Response Relationship, Drug ; Rats

OBJECTIVETo investigate the effects of ethyl acetate extract of Huanglian Jiedu decoction (EAHD) , alone and in combination with fluconazole (FLZ) on FLZ-resistant Candida albicans.

METHODThe minimum inhibitory concentrations (MIC) and sessile MIC80 (SMIC80) of EAHD and FLZ to FLZ-resistant C. albicans were determined by CLSI M27-A3 microdilution method, and the synergy of EAHD combined with FLZ were examined by the checkerboard microdilution assay. Agar plate-method was adopted to observe the rate of antifungal activity according to time-kill curve. HPLC and qRT-PCR were utilized to evaluate the changes of ergosterol content and expressions of related genes, respectively.

RESULTMICs of EAHD ranged from 156 to 1,250 mg · L(-1), those of FLZ from 256 to above 2,048 mg · L(-1) with FICI approximate 0.066 in combination; SMIC80 of EAHD were higher than 1,250 mg · L(-1), SMIC80 of FLZ were higher than 512 mg · L(-1) and up to above 2,048 mg · L(-1). Combination group also showed synergy effect except one group showing addition effect. The results of T-K experiment also confirmed obviously fungicidal effect when treated for 12 h. When compared with control groups, the ergosterol was reduced 85% and 50% in the treatments of combination and EAHD alone by HPLC, respective- ly. The expressions of ERG1, ERG2, ERG6, ERG7 and ERG11 were upregulated, and ACS1, ACS2, MET6 were downregulated when exposed to FLZ. The expressions of the above genes were downregulated by treatment of EAHD. The expressions of ERG2, ERG6, ERG11 were upregulated, while ERG1, ERG7, ACS1, ACS2, MET6 were downregulated in combination group.

CONCLUSIONThe combination of EAHD and FLZ exhibited synergy against FLZ-resistant C. albicans through decreasing the synthesis of ergosterol, and resulting in the breakage of cell membrane.

Antifungal Agents ; pharmacology ; Candida albicans ; drug effects ; growth & development ; metabolism ; Drug Resistance, Fungal ; drug effects ; Drug Synergism ; Drugs, Chinese Herbal ; pharmacology ; Ergosterol ; biosynthesis ; Fluconazole ; pharmacology ; Microbial Sensitivity Tests