Objective To explore the efficacy of intraductal chilled saline perfusion (ICSP) to reduce the thermal bile duct injury during the treatment of radiofrequency ablation (RFA) associating with occlusion of hepatic blood supply in rabbits.Methods 16 healthy New Zealand white rabbits were divided into 2 groups.Rabbits of the ICSP group were placed tubes in the common bile duct after laparotomy,and ICSP was performed during the RFA procedure accompanied with hepatic blood occlusion.While for rabbits of the non-ICSP group,hepatic blood occlusion and RFA were performed without ICSP.RFA electrodes were placed about 5 mm away from the hilus hepatis approximately.Contrast-enhanced ultrasonography (CEUS) was administrated to evaluate the sizes of the ablative zones after the procedure.On post-procedure 6 week,ultrasonography was prerformed to evaluate the changes of the biliary structure,and liver specimens of rabbits wcrc obtained for histopathologic observation of main bile ducts.Results Post-procedure CEUS examination showed that there was no significant difference in the size of the ablative zone between the groups (P ＞0.05).On post-procedure 6 week,rabbits of the ICSP group appeared with biliary dilatation more frequently by ultrasonography (P ＜0.05),and a higher degree of the injury of main bile duct by histopathologic observation (P ＜0.05).Conclusions In treatment of RFA accompanied with hepatic blood occlusion,RFA-induced bile duct injury may be decreased significantly with ICSP.

Objective To establish a method to simultaneously determine contents of alantolactone and isoalantolactone in Sanwei Ganlusan. Methods Contents of alantolactone and isoalantolactone was determined by HPLC was used with PDAD detector;the column was CAPCELL PAK MG C18 (4.6 mm×250 mm, 5μm);the mobile phase consisted of acetonitrile-0.4%phosphoric (58∶42);the flow rate was 1.0 mL/min;the detection wavelength was set at 225 nm;the column temperature was maintained at 30 ℃. Results The linear range of alantolactone was 0.083-0.517 μg (r=0.999 9), and isoalantolactone was 0.108-0.672 μg (r=0.999 9). The mean recovery of alantolactone was 97.95%, RSD=1.31%. The mean recovery of isoalantolactone was 97.69%, RSD=1.24%. Conclusion The method is accurate and simple in operation, which can be used to simultaneously determine contents of alantolactone and isoalantolactone in Sanwei Ganlusan.

Objective To observe the changes of the intercellular spaces of squamous epithelium of lower esophagus in gastroesophageal reflux disease(GERD).Methods Eleven outpatients with GERD symptoms more than 3 months [6 with nonerosive reflex disease(NERD)and 5 with erosive esophagitis(EE)]and 5 healthy volunteers were recruited.All of them underwent endoscopy and 24-hr ambulatory pH monitoring.Biopsies were taken in lower esophagus(2 cm above Z-line)for electron microscope examination.Results Intercellular spaces of esophageal epithelial cell in volun teers,NERD patients and EE patients were (0.374?0.073)?m,(1.308?0.079)?m and (1.332?0.144)?m respectively,with significant differences between the control group and the NERD or EE group.There was no difference between NERD group and EE group.Conclusions Dilated intercellular spaces were seen in both NERD and EE cases,which was significantly different from the control cases.

Dengue virus (DENV) envelope [E] protein is the major surface protein of the virions that indued neutralizing antibodies. The domain III of envelope protein (EDIII) is an immunogenic region that holds potential for the development of vaccines; however, the epitopes of DENV EDIII, especially neutralizing B-cell linear epitopes, have not been comprehensively mapped. We mapped neutralizing B-cell linear epitopes on DENV-1 EDIII using 27 monoclonal antibodies against DENV-1 EDIII proteins from mice immunized with the DENV-1 EDIII. Epitope recognition analysis was performed using two set of sequential overlapping peptides (16m and 12m) that spanned the entire EDIII protein from DENV-1, respectively. This strategy identified a DENV-1 type- specific and a group-specific neutralizing epitope, which were highly conserved among isolates of DENV-1 and the four DENV serotypes and located at two regions from DENV-1 E, namely amino acid residues 309-320 and 381-392(aa 309-320 and 381-392), respectively. aa310 -319(310KEVAETQHGT319)was similar among the four DENV serotypes and contact residues on aa 309 -320 from E protein were defined and found that substitution of residues E309 , V312, A313 and V320 in DENV-2, -3, -4 isolates were antigenically silent. We also identified a DENV-1 type-specific strain-restricted neutralizing epitope, which was located at the region from DENV-1 E, namely amino acid residues 329-348 . These novel type- and group-specific B-cell epitopes of DENV EDIII may aid help us elucidate the dengue pathogenesis and accelerate vaccine design.
Amino Acid Sequence ; Animals ; Antibodies, Neutralizing ; immunology ; Dengue ; virology ; Dengue Virus ; chemistry ; genetics ; immunology ; Epitope Mapping ; Epitopes, B-Lymphocyte ; chemistry ; genetics ; immunology ; Humans ; Mice ; Molecular Sequence Data ; Viral Envelope Proteins ; chemistry ; genetics ; immunology

BACKGROUND: Ephedra, a Chinese medicine, is often used to treat obesity with relatively satisfying results recently. However, the effects of Ephedra on the perimenopausal and postmenopausal obese women remain unclear.OBJECTIVE: To observe the effects of oral Ephedra decoction on body mass and the levels of blood lipids, blood glucose and hormone in ovarietomized obese rats.DESIGN: A completely randomized and controlled experiment.SETTING: Institute of Physiology and Psychology, School of Basic Medical Sciences, Lanzhou University.MATERIALS: The experiment was performed in the Key Laboratory of Pre-clinical Study for New Drugs of Gansu Province and the Laboratory of Institute of Physiology and Psychology, School of Basic Medical Sciences,Lanzhou University from February 2006 to June 2006. Forty-four healthy female SD rats were randomly divided into four groups with 11 rats in each group, namely sham-operated group, ovariectomized group, estrogen replacement therapy group and Ephedra group.METHODS: ① After having been narcotized by cloraminone (110 mg/kg),rats were underwent a bilateral ovariectomy except those in the sham-operated group, which were also operated, but their ovaries were not cut off. ②Rats in the sham-operated group and ovariectomized group were subcutaneously injected with sesame oil (0.2 mL/each rat) every day postoperatively till the end of the experiment. ③ The rats in the estrogen replacement therapy group were given estradiol (1 mg/kg) by subcutaneous injection every day postoperatively till the end of the experiment. ④ The rats in the Ephedra group freely drank 1% water extracts from Ephedra postoperatively, later the concentration of Ephedra gradually increased to 8% on the sixth day, which lasted until the end of the experiment. ⑤ The food intake was monitored daily, and body mass was measured every ten days. ⑥ At the end of the experiment, all the rats were fasted for 12 hours and collected blood samples for the measurement of serum indexes. The body mass and body length were measured to calculate the Lee's index [(g)×103/body length (cm)] at the same time.MAIN OUTCOME MEASURES: ① Body mass and Lee's index at different time points in each group. ② Food intake at different time points in each group. ③ Levels of blood lipids and blood glucose in each group. ④Levels of estrogen, progesterone and insulin in each group.RESULTS: Forty-four rats all entered the analysis of results. ① Result of body mass and Lee's index at different time points: The body masses on the 20th, 30th, 40th and 50th days in the ovariectomized group were (256.4±14.3),(271.3±16.1), (276.4±12.7), (285.7±24.2) g, which were significantly higher than those in the sham-operated group [(226.5±11.5), (241.8±12.6),(243.1±13.5), (251.1±22.4) g, P ＜ 0.05-0.01], and the Lee's index in the ovariectomized group was greater than that in the sham-operated group(317.2±13.5, 280.4±11.2, P ＜ 0.01). The body masses on the 40th and 50th days in the estrogen replacement therapy group were (243.7±14.8) and(246.2±11.9) g, which were significantly lower than those in the ovariectomized group (P ＜ 0.05-0.01), and the Lee's index (289.9±13.5) was lower than that in the ovariectomized group (P ＜ 0.01). The body masses on the 40th and 50th days in the Ephedra group were (245.4 ±14.1) and(252.4±14.9) g, and the Lee's index was 294.4±11.0, which were all lower than those in the ovariectomized group (P ＜ 0.05). ② Result of Food in take at different time points: The food intakes on the 30th, 40th and 50th days in the Ephedra group were (17.8±2.4), (22.3±3.9), (26.1±3.5) g per day,which were decreased as compared with those in the ovariectomized group[(25.9±4.7), (28.5±5.3), (32.8±5.5) g per day, P ＜ 0.05]. ③ Levels of blood lipids and blood glucose: The levels of triglyceride, cholesterol and low density lipoprotein cholesterol (LDL-C) in the ovariectomized group were (1.73±0.32), (1.45±0.50), (0.78±0.19) mmol/L, which were higher than those in the sham-operated group [(0.94±0.29), (1.05±0.30), (0.08±0.11) mmol/L, P ＜ 0.01]. After the estrogen replacement therapy, the levels of triglyceride, cholesterol, LDL-C and blood glucose were (1.10±0.34),(1.14±0.30), (0.17±0.05), (5.88±1.21) mmol/L, which were lower than those in the ovariectomized group (P ＜ 0.05-0.01), but the level of high density lipoprotein cholesterol (HDL-C) was higher than that in the ovariectomized group [(1.11±0.31), (0.88±0.21) mmol/L, P ＜ 0.05]. The levels of triglyceride, cholesterol, LDL-C and HDL-C in the Ephedra group were (0.97±0.16), (1.11±0.20), (0.59±0.07) and (0.45±0.061) mmol/L, which were lower than those in the ovariectomized group (P ＜ 0.05-0.01). ④ The serum levels of estrogen, progesterone and insulin in each group: The serum levels of estrogen and progesterone in the ovariectomized group were lower than those in the sham-operated group [(17.09±9.00), (28.51 ±7.99) μg/L;(58.69±12.11), (62.73±10.93) μg/L, P ＜ 0.01], the serum level of insulin was higher than that in the sham-operated group [(31.74±6.69),(23.75±6.66) mU/L, P ＜ 0.01]. The serum levels of estrogen in the estro gen replacement therapy and Ephedra group were (36.03±8.83) and (30.18±8.61) ng/L, which were higher than those in the ovariectomized group(P ＜ 0.05-0.01), the level of insulin were (21.34±4.57), (24.86±6.20) mU/L,which were lower than those in the ovariectomized group (P ＜ 0.05-0.01).The serum level of progesterone in the Ephedra group [(17.68±6.19) μg/L]was lower than that in the ovariectomized group (P ＜ 0.01).CONCLUSION: Ephedra can promote loss of body mass, reduce levels of the blood lipids and insulin, and increase the serum levels of hormones in ovariectomized obese rats.

Objective To discuss the clinical significance of serum neutrophil gelatinase associated lipocalin(NGAL)detection in the diagnosis of contrast induced nephropathy .Methods A total of 299 inpatients with contrast medium in department of inten‐sive care unit(ICU) ,department of cardiovascular disease ,department of urology from Guangdong Province Hospital of Traditional Chinese Medicine were enrolled ,and the fasting blood glucose and lipids were detected .Sera were collected before examination of contrast medium and 1 ,2 and 6 d after examination of contrast medium respectively .Serum creatinine(SCr)and cystatin C(CysC) were detected ,the concentration of NGAL was determined .CIN was defined as of increased by 44 μmol/L or 25% from baseline in SCr within 24-48 h .The NGAL detection variation tendency was analyzed in different time point .Results In 299 cases ,CIN oc‐curred in 28 patients ,the incidence rate was 9 .36% .The diabetes patients in the incidence of CIN was 16 .21% (12/74) ,no inci‐dence of diabetic patients was 7 .11% (16/225) .When compared with that of before contrast ,CIN in serum of patients with Cr and CysC coronary angiography in 2 d significantly increased after contrast medium ,returned to the level before angiography on 6 d .But the serum NGAL in angiography 1 d after it was significantly increased (P<0 .05) ,statistically significant differences compared with pre contrast ,and was still at a high level on sixth day .Conclusion Prediction of CIN level of serum NGAL in contrast after 1 d could be a good predictor for CIN ,and the prediction time is earlier than the serum levels of Cr and CysC .The level of serum NGAL can be used for early diagnosis of CIN .

Objective To construct a recombinant fusion protein with pneumococcal surface pro-tein A (PspA) of Stretococcus pneumonia (SPN) familyⅠclade 1 and 2, and to analyze the immunogenici-ty of the fusion protein.Methods The gene fragments encoding theα-helix of PspA of the two clades were amplified by PCR and then inserted into the expression vector pET-27b(+) to construct the recombinant ex-pression plasmid.The transformed Escherichia coli BL21 strains carrying expression plasmid were induced by IPTG to express the recombinant protein.The titers and affinity of antibodies against PspA protein were measured by ELISA.An opsonophagocytic assay and an animal experiment were performed to evaluate the immunogenicity of the recombinant protein.Results Double enzyme cutting and gene sequencing confirmed the two purpose gene fragments were correctly expressed in the expression vector pET-27b(+).The titers of anti-PspA antibody in the serum of Kunming ( KM) mice immunized with the fusion protein were 1 ×104 . The affinity of anti-PspA antibody reached to 2×105 .The rates of recombinant PspA6B-PspA05 protein me-diated phagocytosis for SPN6B, SPN05 and SPN01 strains were 20%, 15% and 8.8%, respectively.No SPN23F strain was engulfed by macrophages upon the stimulation with PspA6B-PspA05 protein.The survival rates of mice injected with SPN05, SPN6B, SPN01 and SPN23F strains were respectively 75%, 92%, 75%and 33%upon the immunization of PspA6B-PspA05 protein.Conclusion The recombinant fusion protein PspA6B-PspA05, constructed with the PspA proteins of Stretococcus pneumonia familyⅠclade 1 and 2, was successfully expressed in the E.coli prokaryotic system with the advantage of high immunogenicity.High ti-ters of anti-PspA antibodies with high specificity were induced in KM mice upon the stimulation with Ps-pA6B-PspA05 protein.Moreover, a cross-protective immunity was induced in KM mice upon the immuniza-tion with PspA6B-PspA05 protein.

Objective To study the predictive value of preoperative blood CD4 +/CD8 + ratio in postoperative recurrence after hepatectomy for patients with hepatocellular carcinoma.Methods The clinical data of 67 patients who underwent hepatectomy for hepatocellular carcinoma at The Fifth Hospital Affiliated to Sun Yat-sen University were analyzed retrospectively.Using the preoperative blood CD4 +/CD8 + ratio,these patients were divided into 2 groups,the CD4 +/CD8 + ＜ 1 group and the CD4 +/CD8 + ≥ 1 group.These patients were followed up at the outpatient clinic and/or by telephone till June 2013.The Cox ratio risk pattern analysis was used to determine the significant risk factors of tumor recurrence.Results On univariate analysis,preoperative AFP ≥400 μg/L,maximum diameter of tumor ＞ 5 cm,number of tumor ＞ 3,CD4 +/CD8 + ＜ 1,vascular invasion,positive resection margin,and portal vein tumor thrombus were risk factors of poor disease-free survival (P ＜ 0.05).On multivariant analysis,CD4 +/CD8 + ＜ 1,number of tumor ＞ 3,and portal vein tumor thrombus were independent predictors of poor disease-free survival after hepatectomy for hepatocellular carcinoma (P ＜ 0.05).Conclusions Preoperative CD4 +/CD8 + ＜ 1 was an independent adverse predictor of poor disease-free survival.It was valuable in predicting postoperative recurrence of hepatocellular carcinoma.