Objective:To establish an HPLC method for the determination of chlorogenic acid in honeysuckle stem and honeysuck-le from different sources in different harvest periods. Methods:A Waters C18 column(250 mm × 4. 6 mm,5 μm)was used. The mo-bile phase consisted of acetonitrile-0. 4% H3 PO4(10:90)with the flow rate of 1. 0 ml·min-1 . The detection wavelength was 327 nm and the column temperature was 30℃. An external standard method was established for the determination of chlorogenic acid in honey-suckle stem and honeysuckle in six different harvest periods from three sources. Results:There was a good linear relationship within the range of 0. 065-1. 300 μg for chlorogenic acid(r=0. 999 8). The content of chlorogenic acid in honeysuckle was the highest in Sep-tember and October. The content of chlorogenic acid in Lonicera acuminata was the highest among the honeysuckle stem from three dif-ferent sources. Conclusion:The content of chlorogenic acid in honeysuckle has a certain relationship with the harvest time,which can provide theoretical basis for the choice of harvest time for honeysuckle stem.

OBJECTIVETo study the effect of ethyl acetate fraction of Blumea balsamifera (BBE) residue on treating rats of adjuvant arthritis (AA) and its mechanism.

METHODThe rats were immunized with the Freund's complete adjuvant (FCA). After modeling, 28 days' treatment with BBE was performed. During the experimental process, rat mass, toe girth, arthritic index (AI), proliferation of immune organs and pathological section were measured. After treatment, blood samples were collected through fossa orbitalis vein for detection of serum SOD, MDA, GSH, NO, OH*, ALP, AST, ALT, NAG and SA content using colorimetric method and IL-1, IL-6, TNF-α content using ELISA method.

RESULTAdministration with BBE (high dose) could significantly ameliorate joint swelling and arthritis index, effectively inhibit synovial hyperplasia, down-regulate the levels of MDA, NO, OH*, ALP, AST, ALT, NAG, SA, IL-1, IL-6, TNF-α and up-regulate the SOD and GSH levels in serum.

CONCLUSIONThe results suggested BBE possesses substantial anti-arthritis and antioxidant activities.

Acetates ; Animals ; Anti-Inflammatory Agents, Non-Steroidal ; administration & dosage ; isolation & purification ; Arthritis, Experimental ; blood ; drug therapy ; metabolism ; Asteraceae ; chemistry ; Drugs, Chinese Herbal ; administration & dosage ; isolation & purification ; Humans ; Interleukin-1 ; blood ; Interleukin-6 ; blood ; Male ; Malondialdehyde ; blood ; Oxidative Stress ; drug effects ; Rats ; Rats, Sprague-Dawley ; Tumor Necrosis Factor-alpha ; blood

Background Light-induced retinal damage models vary as many influence factors,herein the modeling method is difficult to copy.It is necessary to establish the grading standardization of retinal damage after retinal light exposure.Objective This study was to improve the modeling method and establish a grading standardization for light-induced retinal damage in rat.Methods Twenty-four SPF 8-10 week-old male SD rats were randomly divided into 4 groups and 6 eyes for each group.The rats were exposed to light intense of 5000 lx for 1,2,3 hours respectively in 3 groups,and other 6 rats served as the normal group.Full-field light exposure experiment was performed for each individual rat separately,and an annular illumination box was used tO ensure the experimental rat moving in a single direction and exposing the right eye in 5000 lx light surrounding during experimental duration.Ganzfeid electroretinogram(ERG)was recorded from the experimental rats at the fifth day after light exposure,and the animals were then sacrificed for histopathology observation to evaluate the retinal thickness change.All procedures which involved animals adhered to the ARVO Statement for the Use of Animals in Ophthalmic and Vision Research.Results After exposing to intensity light for 1,2,3 hours,the b-wave amplitudes of rod response,maximal mixed response,oscillatory potential in scotopic ERG as well as cone response,20 Hz flicker response of photopic ERG were significant declined as lapse of light exposure time(F=71.690,P=0.000；F=56.250,P=0.000；F=23.610,P=0.000：F=27.130,P=0.000；F=27.030,P=0.000)and lowed by 26.2％,52.5％,70.7％,24.4％,39.3％,58.1％respectively at the end of experiment.Meanwhile,the b-wave latencies of rod response,maximal mixed response in scotopic ERG as well as cone response of photopic ERG were evidently different among different groups (F=1.370,P=0.282；F：0.800,P=0.508；F=11.840,P=0.000；F=2.080,P=0.136).Light induced retinal damage located mainly at the temporal retina area.After intensity light exposure for 1,2,3 hours,the thickness of outer nuclear layer at the superior temporal retina attenuated by 11.3％,25.6％and 72.5％,respectively(P<0.05).A significant difference was seen in mean thickness of outer nuclear layer at superior temporal retina among different groups(F=410.27,P=0.000). Conclusion A standardized grading method for light-induced retinal damage is recommended.The continuous illumination in a intensity of 5000 Ix for 1,2,3 hours can induce the mild,moderate or severe retinal damage respectively at temporal retina.

Objective: To investigate the inhibitory effect of Rh2 on HepG2 cells and explore the underlying mechanism.Methods: We used lentivirus carrying β-catenin to infect HepG2 cell, and detected expression of β-catenin using fluorescence microscopy.The effect of Rh2 on proliferation of HepG2-β-catenin and HepG2 cells was measured by CKK-8 assay,and flow cytometry was used to detect cell cycle and apoptosis.The activity of Gsk-3βwas checked by ELISA kit.The expression of Gsk-3β,β-catenin,Bax,Bcl2,CyclinD1,MMP3 genes were measured by qRT-PCR.In order to checked the relationship between β-catenin and TCF4,CHIP assay kit was used,the expression of Bax,Bcl2,CyclinD1,MMP3 genes were measured by PCR.The expressions of Gsk-3β,β-catenin,Bax,Bcl2,CyclinD1,MMP3 proteins were examined by Western blot.Results:HepG2 cells were successfully infected by pLOV-EF1a-MCS-3FLAG-β-catenin lentivirus,named HepG2-β-catenin.CCK-8 showed that ginsenoside Rh2 could effectively inhibit the proliferation of HepG2 and HepG2-β-catenin cells in vitro,which exhibits a dose-dependent manner at range of 10-160 μmol/L Rh2.The IC50 of Rh2 exposure on HepG2 cell for 48,72 h were 100 μmol/L and 58.12 μmol/L,but the IC50 of Rh2 exposure on HepG2-β-catenin for 48,72 h were 129.2 μmol/L,83.33 μmol/L,respectively.The IC50 of Rh2 exposure on HepG2-β-catenin cell was higher than HepG2 cell, compared with HepG2 group the differences was statistically significant ( P<0.01 ).Flow cytometry indicated that Rh2 could arrest HepG2 and HepG2-β-catenin cells in G0/G1 phase;the cell population in G0/G1 phase of HepG2+Rh2 group was(64.57±0.65)%,HepG2-β-catenin+Rh2 group was(58.61±2.01)%.Flow cytometry indicated that Rh2 could induced early apoptosis in HepG2 and HepG2-β-catenin cells.The apoptosis rate of HepG2 +Rh2 group was (17.27 ±2.77)%,HepG2-β-catenin +Rh2 group(9.02 ±1.76)%.The ELISA results indicated that HepG2 cells was induced by Rh2 for 12,24,48,72 h,the activity of Gsk-3βgradually increased,peak in 48 h,then decreased.Compared with control group,Rh2 induced HepG2 and HepG2-β-catenin cells for 48 hours, Gsk-3βactivity were increased, and their activity reduced after adding Bio, there were no significant differences between HepG2+Rh2 and HepG2-β-catenin+Rh2 groups.The PCR,CHIP and WB results showed that the expression of Gsk-3β,Bax gene and proteins increased,while theβ-catenin,CyclinD1,Bcl2,MMP3 gene and proteins down-regulation in HepG2 and HepG2-β-catenin cell induced by Rh2.Compared with HepG2-β-catenin +Rh2 group, the expression of other gene and proteins changed significantly,however,Gsk-3βwas no significant difference.Conclusion:Over-expression of β-catenin may weaken the phar-macological effects of ginsenoside Rh2 on HepG2 cells.The activity of Gsk-3βwas increased by ginsenoside Rh2 to degradeβ-catenin, affecting the expression of downstream genes,promoting apoptosis of liver cancer cells and inhibiting metastasis.

Objective:To study the mechanism of ginsenoside Rh2 inhibiting HepG2 cells migration.Methods:HepG2 cells in logarithmic growth phase were cultured in 96-well plates,which were induced by different concentration Rh2,respectively for 24,48,72 hours.The cell inhibition was detected by Cell Counting Kit.Transwell chambers was used to checked HepG2 cell migration ability;luciferase was tested by Luciferase Reporter Assay system reagent;The expressions of P-ERK,ERK,P-P38,P-38,P-JUK,JUK,MMP3 proteins were detect by Western blot;the expression of AP1,MMP3 gene were detected by Quantitative PCR;The expression of AP1, MMP3 fluorescence protein were observed by fluorescence microscopy.Results:Administrated with different concentration of Rh2 after 24 ,48 ,72 h,the proliferation of HepG2 cells were inhibited ( P<0.05) ,and in dose-and time-dependent manner.Transwell assay showed Rh2 could significantly inhibited migration of HepG2 cells.The expressions of P-ERK , MMP3 proteins were significantly decreased,the expressions of P-JUK, P-P38 proteins were significantly increased, expression levels of ERK, P-38, JUK were no significant difference.Expression of AP1,MMP3 gene were significantly decreased,the expressions of AP1,MMP3 fluorescence proteins were significantly decreased.Conclusion:Ginsenoside Rh2 can activate MAPK pathway to inhibit the migration of HepG2 cells.

Objective To study the indication,feasibility and short-term efficacy of combined thoracoscopic and laparoscopic radical esophagectomy for the treatment of esophageal cancer.Methods Retrospective medical records analysis was conducted for 139 esophageal cancer patients who underwent combined thoracoscopic and laparoscopic esophagectomy in our department from December 2009 to August 2011.The tumors were located in upper esophagus in 16 cases,middle esophagus in 107 cases,and lower esophagus in 16 cases.The surgery started with the thoracoscopic mobilization of thoracic esophagus and lymph nodes dissection,which were followed by the laparoscopic stomach mobilization and gastroesophageal anastomosis in left neck.Postoperative pathological staging identified stage Ⅰ esophageal cancer in 25 cases ( stage Ⅰ a:13 cases,stage Ⅰ b:12 cases),stage Ⅱ esophageal cancer in 71 cases,stage Ⅲ esophageal cancer in 31 cases ( stage Ⅲ a:16 cases,stage Ⅲ b:15 cases) and stage Ⅳ esophageal cancer in 12 cases.Results Except for open conversions in 4 cases (2.9％),all surgical operations were completed smoothly.Postoperative anastomotic leak was found in 6 cases(4.3％ ),chylothorax in 1 case(0.7％ ),arrhythmia in 4 cases(2.9％ ),and dumping syndrome in 1 case( 0.7％ ).All of these complicated cases fully recovered after conservative treatments.Postoperative lung infection was found 11 cases (7.9％),3 of whom required tracheotomy and assisted ventilation and 1 case died as a result of the infection (mortality rate:0.7％ ).Ten cases(7.2％ ) presented with hoarseness postoperatively.Out of the 139 cases,130 cases were successfully followed up with durations ranged from 1 to 20 months,during of time the esophageal cancer spread to liver in 2 cases,celiac lymph nodes in 4 cases,lung in 2 cases,and bone in 1 case.Ten cases died,and all remaining cases remained alive during the follow up.The one-year survival rate was 88.9％ for these cases.Conclusion Combined thoracoscopic and laparoscopic radical esophagectomy is a technically safe and feasible treatment for esophageal cancer.The short-term efficacy results are satisfactory.This technique is indicated not only for early and middle stage esophageal cancer,but also for some of the advanced esophageal cancer cases.

Objective To investigate the adrenal steroidogenic function in genotype-proven heterozygotes carrying mutations in CYP17A1 gene in vivo. Methods Eight patients and 14 family members from 5 families with 17-hydroxylase/17,20-lyase deficiency (17OHD) were recruited. The mutations of the CYP17A1 gene in these individuals were screened by direct sequencing of PCR products. The hormonal response to ACTH was evaluated in the 14 genotype-proven carriers and 45 age- and sex-matched normal subjects. Results Three mutations were found in 5 unrelated families. 14 carriers with CYP17A1 mutation were identified, including 7 heterozygotes with D487_F489del, 6 with Y329fs, and 1 for H373L. Compared to the normal subjects, the carriers exhibited lower basal and ACTH-stimulated cortisol levels, but higher ACTH-stimulated corticosterone level. The ratios of corticosterone to cortisol in the genotype-proven heterozygotes were higher than those of normal individuals at baseline and following ACTH-stimulation. Similarly, progesterone level and ratios of progesterone to 17-hydroxyprogesterone in the male heterozygotes were also higher than that of normal individuals before and after stimulation. No significant differences were observed in the hormone levels between two genotypes (D487_F489del vs Y329fs). Conclusions Genotype-proven carriers of 17OHD without apparent clinical symptoms exhibit decreased enzyme activity,analogous to mildly impaired adrenal 21-hydroxylase activity in the carriers of CYP21 A2 gene mutation.

Objective To investigate the effects of mercury on T lymphocytes and serum immune indexes of workers with Methods occupational mercury exposure. A total of 45 workers with occupational mercury exposure were selected as the ， mercury exposure group and 47 workers without occupational mercury exposure were selected as the control group using the judgment sampling method. Cold atomic absorption spectrometry was used to detect the urinary mercury level of the two groups. （ ） +， + +， + + - + Flow cytometry was used to detect the proportion of cluster of differentiation CD 3 CD3CD4 CD3CD8 and CD3CD19 ， - （ - ） - （ - ） cells in peripheral blood and the levels of tumor necrosis factor α TNF α and interleukin 8 IL 8 in serum. The levels of （ ） ， Results immunoglobulin Ig A IgG and IgM in serum were measured by immune nephelometry. The urinary mercury level of （ ： vs ，P ） individuals in the mercury exposed group was higher than that of the control group median 92.7 13.2 μg/g Cr <0.01 . The +， + +， - + proportion of CD3 CD3CD4 CD3CD19 cells in peripheral blood and serum IgG level in the mercury exposed group （ P ）， - - （ P ） decreased all <0.05 and the serum TNF α and IL 8 levels increased all <0.01 compared with the control group. Urinary - + mercury level was negatively correlated with the proportion of CD3CD19 cells in peripheral blood and serum IgG level in the ［ （r） ， ， P ］， study subjects Spearman correlation coefficient S were −0.21 and −0.31 respectively all <0.05 and positively - - （r ， ， P ） ， correlated with serum TNF α and IL 8 levels S were 0.36 and 0.39 respectively all <0.05 . However the urinary mercury （ P ）， +， + +， level was neither correlated with IgA and IgM levels in serum all >0.05 nor with the proportion of CD3 CD3CD4 + + （ P ） Conclusion CD3CD8 cells in peripheral blood all >0.05 . Occupational exposure to mercury can lead to abnormal ， changes in peripheral blood T lymphocyte subsets B lymphocytes and serum immune factors in workers. The mercury load of occupational mercury exposure workers may impact their immune function.

Objective To explore the correlationships between microperfusion diffusion indexes derived from intravoxel incoherent motion(IVIM)and perfusion values measured by arterial spin labeling (ASL)in renal allograft. Methods A total of 76 renal allograft recipients and 26 age-matched volunteers (group 0)were included in this prospective study. All subjects were underwent conventional MRI, IVIM and ASL MRI which were performed in the oblique-sagittal plane. Seventy-six recipients were divided into two groups based on the estimated glomerular filtration rate(eGFR):recipients with good allograft function(group 1, eGFR≥ 60 ml · min-1 · 1.73m-2,n=44)and recipients with impaired allograft function(group 2, eGFR<60 ml · min-1 · 1.73m-2,n=32). Three IVIM indexes values, including true diffusion coefficient(ADCslow), pseudo-diffusion coef fi cient(ADCfast), perfusion fraction(PF), and one ASL index value of renal cortex(renal blood flow, RBF)were measured. One-way analysis of variance and the least significant difference were used to compare the different of each cortical index values among three groups. Correlations between the ADCslow, ADCfast, PF, RBF and eGFR as well as the correlation among the indexes were evaluated using Pearson correlation coefficients. Results For cortical ADCslow, ADCfast, PF and RBF values, allografts with good function and impaired function showed significantly differences compared healthy controls(all P<0.01). In allografts with good function, cortical ADCslow,ADCfast,PF showed no significantly differences compared with controls(all P>0.05), but RBF value was significantly lower(P<0.05). The ADCslow, ADCfast, PF and RBF values of renal cortex were significantly lower in allografts with impaired function compared to allografts with good function(all P<0.01). In renal allografts, there were significant positive correlations between cortical ADCslow, ADCfast, PF, RBF value and eGFR(r values were 0.604, 0.552, 0.579 and 0.673, all P<0.01). Cortical ADCfast and PF value exhibited a significant correlation with RBF for recipients(r values were 0.501 and 0.423, all P<0.01). Conclusion Cortical ADCfast and PF values derived from IVIM and RBF measured by ASL show a significant positive correlation in renal allografts.

OBJECTIVE: To analyze the echocardiographic abnormalities and the prevalence of anticardiolipin antibodies (aCL) in systemic lupus erythematosus (SLE) patients and to evaluate the relationship between aCL and cardiac valvular abnormalities in SLE patients. METHODS: Ninety SLE patients were performed M-mode, 2-dimensional and Doppler echocardiography and aCL IgG and IgM were measured by an enzyme-linked immunosorbent assay (ELISA). According to the abnormalities in the echocardiography, the patients were assigned into valvular abnormality group and non-valvular abnormality group. Chi-square method was used to compare the difference of aCL prevalence between the two groups. RESULTS: The prevalence of echocardiographic abnormalities was 53.33%, and valvular abnormality (38.89%) and pericardial effusion (34.44%) presented most frequently. The aCL prevalence was 32.56% in the 43 SLE patients. The prevalence of aCL in the valvular abnormality group was significantly higher than that in non-valvular abnormality group (52.94% vs 19.23%, P<0.05). CONCLUSION The incidence of echocardiographic abnormalities is high in SLE patients, most often in valves and pericardium. The aCL is probably related to valvular damage in SLE patients.
Adolescent ; Adult ; Antibodies, Anticardiolipin ; blood ; Echocardiography, Doppler ; Female ; Heart Valve Diseases ; etiology ; immunology ; physiopathology ; Humans ; Lupus Erythematosus, Systemic ; complications ; immunology ; physiopathology ; Male