OBJECTIVE:To establish an HPLC method for simultaneous determination of Naringin,Hesperidin and Neohesperidin in the effective fraction of flavone in Fructus Aurantii Immaturus.METHODS:Hypersil C18 column(250 mm?4.6 mm,5 ?m) was used with the mobile phase consisted of acetonitrile -0.1% phosphoric acid solution(20∶80) at a flow rate of 1.0 mL?min-1,and the detection wavelength was set at 283 nm and the column temperature was maintained at 30 ℃.RESULTS:The calibration curves of Naringin,Hesperidin and Neohesperidin were in good linearity over the ranges of 0.44～2.20 ?g(r=0.999 5),0.025 6～0.128 0 ?g(r=0.999 3),0.54 ～ 2.70 ?g(r=0.999 9),respectively.And the average recoveries for the three constituents were 99.41%,100.33% and 99.69%,respectively with RSD at 1.14%,1.47%,and 1.16%,respectively.CONCLUSION:The method is simple,accurate and reliable,and can be used for the quality control of Fructus Aurantii Immaturus.

BackgroundThere are a lot of studies about the wound healing charateristics of cornea after SBK with femtosecond laser.In our study,mechanical microkeratome was preferred for corneal flap.We observed the proliferation of keratocytes by investigating the myofibroblast ( MF) activity.Objective The study was to compare the morphologic and histological changes in the cornea after sub-Bowmans keratomileusis ( SBK) with photorefractive keratectomy ( PRK) and laser in situ keratomileusis ( LASIK)by investigating the express of transforming growth factor-β( TGF-β)and α-smooth muscle actin( α-SMA)and to investigate the wound healing characteristiCs of cornea after SBK.MethodsTwenty-seven adult New Zealand white rabbits were randomly assigned into group A,B and C.SBK waa performed on the right eyes of each rabbit in group A,LASIK for group B and PRK for group C.All the left eyes were used as the normal control group.Histological examinations by light microscopy were performed on day 7,1 months,3 months after surgery.The expression of α-SMA and TGF-β and the number of activated MF were assessed by immunohistochemiatry.ResultsIn SBK group,corneal epithelium cells proliferation around the wound was seen and the numbers of active fibroblasts were increased after surgery.The expression of α-SMA or TGF-β around the corneal flap and the corneal 8troma started at day 7 postoperatively and peaked at 1 months and decreaaed around the corneal flap and the corneal 8troma started at day 7 postoperatively and peaked at 1 months and decreaaed t3mnh.TFβep( SBK:t=2.226,2.158,2.330,P＜0.05;PRK:t=4.745,6.524,6.293,P＜O.05).The numbersof activ( SBK:t=2.226,2.158,2.330,P＜O.05;PRK:t=4.745,6.524,6.293,P＜0.05).The numbersof activatedMFs were different fromLASIKstatisticallytoobetweenSBKgroupandPRKgroup ( SBK:t =4.439.5.692,4.175,P＜0.05 ; PRK:t=6.330,6.723,5.267,P＜0.05 ).Theα-SMAand TCF-βexpressionsin SBKgroupwerelessthan PRK group but more thanLASIKgroup( TCF-β:t =4.691,5.527,t =4.399,P＜0.05 ; α-SMA:t =9.637.10.282,8.197,P＜0.05).The numhers of MFs in SBK group was less than PRK group before 3 months and were same at 3 months ( t =5.188,4.529,P＜0.05 ).Conclusions ComparedwithconventionalLASIK,SBKcanup-regulatethe expression of α-SMA.TGF-β,activated MFs in the corneal flap.which enhance corneal biomechanics and promote healing.However,most of the disadvantages caused by wound healing in SBK still remain compared to PRK.

Objective To study the relationship between coronary artery disease (CHD ) and interleukin-6 (IL-6), TNF-α and IL-10. Methods Patients with CHD were included in CHD group (n=628) and patients without CHD con?firmed by coronary angiography were selected in control group (n=540) . The recorded data included age, body mass index (BMI) and serum levels of glucose (GLU), total cholesterol (TC), triglyceride (TG), high density lipoprotein-cholesterol (HDL-C) and low density lipoprotein-cholesterol (LDL-C). Meanwhile serum levels of IL-6, TNF-α, IL-10 were determined by ELISA. Results Serum levels of BMI, LDL-C, TG were higher in CHD group than those in control group(P<0.01). The serum level of HDL-C was significantly lower in CHD group than that in control group(P<0.01). Serum levels of IL-6(P<0.05)and TNF-α(P<0.01)were higher in CHD group than those in control group. The serum level of IL-10 was significant?ly lower in CHD group than that in control group(P<0.01). Conclusion IL-6 and TNF-αare involved in the develop?ment of CHD. IL-10 can inhibit inflammation and protect vessel integrity.

Objective To observe the lung protective effect of Tongfu Xiefei method (TFXF) in rats with sepsis, and to discuss its possible mechanism.Methods Forty-two Sprague-Dawley (SD) rats were randomly divided into blank control group (n = 6), model group (n = 18) and TFXF group (n = 18). Sepsis model was reproduced by cecal ligation and puncture (CLP) in rats of model group and TFXF group. After the reproduction of sepsis model, rats in TFXF group received Tongfu Xiefei granules 0.01 mL/g by gavge, while those in model group were given equal dose of normal saline by the same way. The rats in blank control group received no treatment. At 3, 6, 12 hours after CLP, abdominal aorta blood was collected for blood gas analysis and inferior vena cava blood was collected for determination of the concentrations of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6). Bronchoalveolar lavage fluid (BALF) was collected for measurement of concentrations of total protein (TP), total phospholipid (TPL), and desaturated phosphatidyl choline (DSPC). The ratio of wet/dry lung weight ratio (W/D) was measured, and malondialdehyde (MDA) and myeloperoxidase (MPO) in lung tissues were determined. The pathologic changes in their lungs were observed with light microscopy.Results Compared with those in blank control group, the levels of pH value, arterial oxygen partial pressure (PaO2), HCO3-, base excess (BE) were lowered, and partial pressure of carbon dioxide of arterial blood (PaCO2) was increased in model group. The serum concentrations of TNF-α and IL-6 were gradually increased after the reproduction of sepsis model. Compared with those in blank control group, the levels of TP, TPL, and DSPC/TPL in model group were decreased, while the levels of W/D, MDA and MPO were increased. Compared with those in model group, pH value was elevated in TFXF group at 3 hours (7.27±0.04 vs. 7.18±0.07,P < 0.05). PaO2 (mmHg, 1 mmHg = 0.133 kPa) was improved at 3, 6, 12 hours (3 hours: 128.00±16.05 vs. 106.78±10.73, 6 hours: 98.46±15.97 vs. 72.80±16.33, 12 hours: 90.70±9.31 vs. 74.28±12.19, allP < 0.05). The serum concentrations of TNF-α (ng/L) in TFXF group were significantly lower than those in model group at 12 hours (508.20±94.08 vs. 756.60±138.77,P < 0.05), and the serum concentrations of IL-6 (ng/L) in TFXF group were significantly lower than those in model group at 6 hours and 12 hours (6 hours: 687.80±35.00 vs. 849.40±148.28, 12 hours: 728.80±214.41 vs. 917.00±245.96, bothP < 0.05). Compared with those of model group, the levels of TP (g/L) in BALF in TFXF group were significantly decreased at 12 hours (1.01±0.23 vs. 1.60±0.47,P < 0.05), and the levels of TPL (mg/L) in TFXF group were significantly increased at 12 hours (86.40±11.33 vs. 62.40±16.33,P < 0.05). The levels of DSPC/TPL in TFXF group were significantly higher than those in model group at 6 hours and 12 hours (6 hours: 0.58±0.13 vs. 0.38±0.10, 12 hours: 0.45±0.13 vs. 0.24±0.07, bothP < 0.05). The levels of W/D in TFXF group were significantly higher than those in model group at 3 hours (3.84±0.25 vs. 2.99±0.50,P < 0.01), but lower than those in model group at 12 hours (3.21±0.53 vs. 4.89±1.14,P < 0.05). The levels of MDA (nmol/mg) in TFXF group were significantly lower than those in model group at 6 hours and 12 hours (6 hours: 4.04±2.58 vs. 8.89±2.61, 12 hours: 11.31±3.60 vs. 20.60±8.10, bothP < 0.05), while the levels of MPO (U/g) in TFXF group were lower than those in model group at 12 hours (4.79±0.66 vs. 7.22±1.76,P < 0.05). Compared with model group, the lungs in TFXF group showed less morphological changes under light microscopy, such as pulmonary edema, congestion, effusion and fibrosis.Conclusions The method of Tongfu Xiefei may improve hypoxemia and metabolic acidosis, alleviate lung edema and ameliorate pulmonary pathological changes in rat sepsis model. Tongfu Xiefei method shows a protective effect in sepsis by the way of reducing peroxidative damage, inhibiting the release of proinflammatory factors and abating degradation of lung surfactant.

Child ; Humans ; Leukemia, Myeloid, Acute ; complications ; Male ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; complications

Objective Comparison of three different molecular assays for the detection and molecular characterization of circulating tumor cells in breast cancer .Methods The retrospective study compared three different molecular assays to detect CTC in the peripheral blood of 30 healthy individuals and 71 benign breast disease patients and 83 early and 84 metastatic breast cancer patients .All samples were collected at the outpatient , inpatient and physical examination department of Sichuan Provincial People ′s Hospital from January 2011 to June 2014.The same cDNAs were analyzed by:singleplex RT-qPCR assay for BCL-2, multiplex RT-qPCR for BCL-2, HER-2, HMAM, and a commercially available molecular assay (AdnaTest BreastCancer ) for GA733-2, MUC-1, HER-2.The positive of CTC were compared among healthy individuals and benign breast disease patients and breast cancer patients .Chi square test was used to compare the expression of gene markers among the three groups , and the agreement of Kappa test was used to evaluate the method.Results (1) Detection rates of early breast cancer by single RT-qPCR, Adna kits and multiple RT-qPCR were 13.3%, 16.9% and 18.1%, respectively , and the detection of metastatic breast cancer were 31.0%, 42.9%and 35.7%, respectively.There were significant differences in the positive of CTC by three molecular assays between healthy individuals and benign breast disease patients and early breast cancer patients ( The test values were 4.235 and 4.301, 5.367 and 5.474, 5.894 and 6.023 respectively, P<0.05).There were no differences between benign breast disease patients and early breast cancer patients (The test values were 0.891,0.748 and 0.701 respectively,P >0.05) .There were significant differences between metastasis breast cancer patients and healthy individuals and benign breast disease patients and early breast cancer patients ( The test values were 8.429,7.553 and 7.061;10.24, 9.025 and 8.745; 9.658, 8.417 and 8.201 respectively,P<0.05).(2) In early breast cancer: The concordance between AdnaTest and single RT-qPCR was 79.5%while between AdnaTest and multiplex RT-qPCR was 77.1%.No agreement was found among them ( The test values were 1.065 and 1.871, P were 0.371 and 0.258 ) .The concordance between single RT-qPCR and multiplex RT-qPCR was 80.7%.No agreement was found between them (The test values was 2.814, P was 0.156).(3) In patients with overt metastasis:The concordance between AdnaTest and single RT-qPCR was 78.6%( The test values was 10.986).While between AdnaTest and multiplex RT-qPCR was 80.9%( The test values was 9.251 ) . Agreements were found among them ( P was 0.002 and 0.005 respectively ) .The concordance between single and multiplex RT-qPCR was 88.1%( The test values was 12.364 ) .Agreement was found between them (P was 0.001).Conclusions No correlations were found among different molecular methods to detect CTC in the early primary breast cancer , but correlations were found in the metastatic breast cancer , suggesting that different rate of CTC caused by the number of CTC and its heterogeneity should be considered to the clinical diagnosis and treatment of breast cancer while molecular method is used .

Objective To investigate the effect of pseudolaric acid B(PLAB) on growth of human gastric carcinoma cells in vitro.Methods The expression of PPAR? was detected by RT-PCR;the effect of PLAB on cell growth was tested by MTT;Hoechst33342/PI and DNA gel electrolysis were employed to examine apoptosis;cell cycle was checked by flow cytometry.Results When treated with 0.1～10 ?mmol/L PLAB for 72,the proliferation of MGC803 cells was significantly inhibited.The proportion of MGC803 cells at G2 phase was significantly increased when treated with 10 ?mmol/L PLAB after 48 h,and showed an apparent G2 phase arrest.After treatement with PLAB for 72,typical apoptotic changes were observed.The expression of PPAR? was at a low level in MGC803 cells and up-regulated when treated with 10 ?mmol/L PLAB for 48 h(P

OBJECTIVETo investigate the effect of Moutan Cortex on mesangial proliferation and basement membrane thickening induced by advanced glycation end products (AGEs).

METHODThe glomerular mesangial cells (MC) injury model was established by inducing by AGEs. The cell were divided into 6 groups: the blank group ( BSA, 200 mg L-1) , the model group (AGEs, 200 mg L-1), the positive control group (AG, 10 mmol L L-1), and drug administration groups, namely the Moutan Cortex-treated high-dose group (2 x 10(-4) g mL(- 1)), the Moutan Cortex-treated medium-dose group (1 x 10(-4) g mL-1 ), and the Moutan Cortex-treated low-dose group (0. 5 x 10(-4) g . mL(-1)). The MTT method was performed to observe the effect of Moutan Cortex on the proliferation of MC. The content of fibronectin (FN) and collagen secretion 1V (Col IV) in cell supernatant were detected by ELISA kits. The western blot analysis was carried out to observe the FN expression. The Real-time PCR analysis was applied to examine the Col IV mRNA expression.

RESULTAGEs significantly increased AGEs-induced MC proliferation and FN and Col 1V secretion. The western blot analysis showed that MC could down-regulate the FN expression of MC secretion. According to the results of the real-time PCR assay, MC could down-regulate AGEs-induced MC secretion Col IV mRNA expression.

CONCLUSIONMC had a certain protective effect on MC cultured under AGEs conditions. MC could remarkably inhibit the composition and secretion of Col IV and FN in matrix and the basement membrane thickening, and provide an experimental basis for the treatment of diabetic nephropathy.

Animals ; Basement Membrane ; drug effects ; metabolism ; Cell Line ; Cell Proliferation ; drug effects ; Collagen Type IV ; genetics ; secretion ; Drugs, Chinese Herbal ; pharmacology ; Fibronectins ; biosynthesis ; Gene Expression Regulation ; drug effects ; Glycation End Products, Advanced ; adverse effects ; Mesangial Cells ; cytology ; drug effects ; metabolism ; secretion ; Paeonia

Objective To study the changes of serum level of high mobility group box-1(HMGB- 1)in patients with multiple trauma in order to forecast organ dysfunction(OD)and deaths.Methods The optical densities of HMGB-1 in serum of 35 patients with multiple trauma were determined on 1st,3rd, and 7th days after trauma,and the incidence of organ dysfunction and deaths were evaluated,then analyzed statistically to learn the relation between the serum levels of HMGB-1 and deaths with an attempt of predic- ting the incident of organ dysfunction and deaths.Results (1)As OD was concerned,there was a statis- tically significant difference in optical density of HMGB-1 on 1st and 3rd days between the two groups of multiple injury patients(t=4.411,P

20(Z= -2.117, P=0.034), and between the patients with OD and without OD (Z=-3.089, P=0.002), but PCT was not so between the non-surviror and survivor (Z=-1.307, P=0.191). The serum PCT level correlated with the incidence of organ dysfunction (x~2=14.82, P=0.033) and APACHEII (x~2=12.83, P