Objective Gambogic acid ( GA) can suppress the growth of multiple tumor cells , including gastric carcinoma , hepatoma , hematologic neoplasms and breast carcinoma , but there have been few reports about its effect on urologic neoplasms .This study was to investigate the possible mechanisms of GA inducing bladder cancer cell apoptosis and cell cycle arrest . Methods We cultured human bladder cancer BIU8-7 cell lines in vitor and treated the cells in the logarithmic growth phase with isotonic saline solu-tion (negative control)or GA at the concentrations of 1.0, 2.0, and 3.0μmol/L, respectively.We determined the expression of the Caspase-3 protein in the tumor tissue using the immunohistochemical S-P method and detected GA-induced apoptosis of the bladder cancer cells and cell cycle changes by flow cytometry . Results The expressions of the Caspase-3 protein were 4.28 ±1.86, 5.03 ± 0.78, and 6.47 ±1.31 in the 1.0, 2.0, and 3.0μmol/L GA groups, respectively, significantly higher than 2.13 ±1.27 in the nega-tive control (P<0.05).Flow cytometry showed a gradual decrease of the cells in the G 0/G1 phase and a gradual increase in the G2/M phase , but no obvious change in the S phase . Conclusion Gambogic acid can promote the apoptosis , arrest the cell cycle , and in-hibit the proliferation of bladder cancer cells by increasing the expression of the Caspase -3 protein.

Objective To investigate the prevalence of clonorchiasis among residents of Huachuan Country,Heilongjiang Province and to provide a basis for development of control strategies.Methods From 2011 to 2012,cluster random sampling was performed to survey the incidence of clonorchiasis in Huachuan Country.Fecal specimens were collected and examined the clonorchis sinensis eggs by Kato-Katz method.A questionnaire survey was conducted to collect related information such as age,gender,occupation and eating habits.The infection characteristic was analyzed.Results Totally 884 patients with clonorchiasis were found among 2248 residents,and the infection rate was 39.32％.The infection rate in male[47.15％(611/1296)] was significantly higher than females [28.68％(273/952),x2 =34.55,P ＜ 0.01].The infection rate increased with age,which was higher in the 20-69 years old people,with the highest infection rate in the 50-59 years old groups[45.34％ (219/483)].Of the occupational distribution,farmers had the highest infection rate [47.24％ (420/889)],followed by cadres and staffs[38.38％(190/495)].Of residents with fresh fish eating history,the prevalence of clonorchiasis was 53.38％(150/281).Conclusions The prevalence of clonorchiasis is still high in Huachuan County.To reduce the prevalence of clonorchiasis,comprehensive prevention measures,health education and group chemotherapy should be carried out.

Objective To investigate the infect capability of recombinant adenovirus mediated PDGF to human umbilical cord derived mesenchymal stem cells (MSCs). Methods The PDGF cDNA sequence was amplified with RT-PCR and constructed recombinant adenovirus vector AdPDGF. The human umbilical cord derived MSCs were isolated and cultured. In vitro AdPDGF infected MSCs with various MOI,and determine the efficiency using FACS and fluorescence microscope. Trypan blue and MTT assay was used to detect cell viability and proliferation. ELISA was used to quantify PDGF secretion. Results Recombinant adenovirus AdPDGF was successfully and constructed,in which infection efficiency to MSCs was dose dependent. The highest efficiency was around 87.36 % when MOI =50,at which cell viability and proliferation wasn' t impaired. Cell viability of uninfected,AdPDGF infected and control virus infected MSCs was (97.8 ±2.3) %,(91.9±4.0) % and (92.8±4.0) %,separately. The cell proliferation was (100±16.8) %,(95.9±12.0) % and (87.5±9.7) %,separately. There was no statistic significance among AdPDGF infected,control virus infected and uninfected MSCs. 48 hrs post infection,PDGF secretion can be measured from infectious MSCs' supernatant. The secretion level was (1.53±0.37),(3.03±0.68) and (5.25±0.92) ng/ml,separately,when MOI= 10,30,50. No PDGF can be detected from MSC-GFP and control MSC group. Conclusion Recombinant adenovirus AdPDGF was constructed and can infect human umbilical cord derived MSC at high efficiency.

An electrochemical L-lactate biosensor was fabricated by combining Platinum nanoparticles (Pt-nano) with multi-walled carbon nanotubes(MWCNTs).L-lactate oxidase(LOD) was immobilized on the surface of the glassy carbon electrode (GCE) modified with MWCNTs and Pt-nano.The surface of resulting LOD/MWCNTs/Pt-nano electrode was covered by a thin layer of sol-gel to avoid the loss of LOD and to improve the anti-interference ability.The cyclic voltammetric results indicated that MWCNTs/Pt-nano catalyst displayed a higher performance than MWCNTs.Under the optimized conditions, i.e., applied potential of 0.5 V, pH 6.4, 25 ℃, the proposed biosensor's determination range was 0.2-2.0 mmol/L, response time was within 5 s, and the sensitivity was 6.36 (A/(mmol/L).It still kept 90% activity after 4 weeks.The fabricated biosensor had practically good selectivity against interferences.The results for whole blood samples analyzed by the present biosensor showed a good agreement with those analyzed by spectrophotometric method.

Nine compounds were isolated from an ethanol extract of the roots of K. roxburghii by using a combination of various chromatographic techniques including column chromatography over silica gel, MCI gel, Sephadex LH-20, and reversed-phase HPLC. On the basis of physical-chemical properties and spectroscopic data analysis, their structures were identified as munjistin (1), 1-methoxy-3,6-dihydroxy-2-hydroxymethyl-9,10-anthraquinone (2), 1,2,3-trihydroxy-9,10-anthraquinone (3), arjunolic acid (4), hyptatic acid-A (5), hyptatic acid-B (6), 2α,3β,24-trihydroxyurs-12-en-28-oic acid (7), 2α,3β,23-trihydroxyurs-12-en-28-oic acid (8), and daucosterol (9). Compounds 1-9 were obtained from this genus for the first time.
Anthraquinones ; chemistry ; isolation & purification ; Rubiaceae ; chemistry ; Triterpenes ; chemistry ; isolation & purification

AIMTo identify the hydroxylate metabolites and its sulfate conjugates of daidzein in rat urine.

METHODSUrine samples from 0 - 24 h were collected after single ig dose of 500 mg x kg(-1) daidzein to each of six rats. The urine samples were purified by SPE column (SPE C18) and analyzed with liquid chromatographic-tandem electrospray ionization ion trap mass spectrometry (LC-ESI/MS(n)) for potential metabolites.

RESULTSSeveral new hydroxylate metabolites and its sulfate conjugates were found and identified in rat urine.

CONCLUSIONLC-ESI/MS(n) is proved to be a simple, rapid, sensitive and specific technique for identification of the hydroxylate metabolites and its sulfate conjugates of daidzein in rat urine.

Animals ; Chromatography, Liquid ; methods ; Hydroxylation ; Isoflavones ; chemistry ; metabolism ; urine ; Male ; Molecular Structure ; Phytoestrogens ; chemistry ; metabolism ; urine ; Rats ; Rats, Sprague-Dawley ; Reproducibility of Results ; Seeds ; chemistry ; Soybeans ; chemistry ; Spectrometry, Mass, Electrospray Ionization ; methods ; Sulfates ; metabolism ; Tandem Mass Spectrometry ; methods

Objective To investigate image quality and clinical value of dual-source dual energy virtual non-contrast (VNC) CT of the head. MethodsSixty-two patients suspected of cerebrovascular diseases underwent conventional non-contrast (CNC) CT and dual energy CTA examination of the head with dual-source CT. Virtual non-contrast images were reconstructed using dual energy software. The CT values of gray matter, white matter, cerebrospinal fluid, hyperdense hemorrhagic lesion and hypodense ischemic lesion were compared between CNC and VNC images. A four-score scale was used to assess image quality subjectively. Image noise, radiation dosage and detection rate were compared between CNC and VNC images. Paired t test, Wilcoxon signed ranks test and Chi-square test (McNemar test and Kappa test) were used. Results The CT value on CNC and VNC images, were (43. 3 ± 1.5) and (33. 2 ± 1.3) HU for gray matter (t = 46.98, P ＜ 0. 01), (32. 9 ± 1.3) and (28.8 ± 1.6) HU for white matter(t = 16. 28, P ＜0.01), (9.0 ± 1.4) and (5.3 ± 1.9) HU for cerebrospinal fluid (t=12.41, P＜0.01),(62.8 ±10.0) and (51.3 ± 11.5) HU for hyperdense lesion (Z = -4.37, P ＜ 0.01), (20.7 ±4.7) and (18.0 ±6. 9) HU for hypodense lesion (t = 3. 84, P ＜ 0. 01), respectively. VNC images[(1.63 ±0.34) HU]had more noise than CNC images[(0.99±0.18) HU](Z= -6.41, P＜0.01). VNC [(0. 53 ± 0. 08) mSv]had less effective dose than CNC[(1.37 ± 0. 23) mSy](Z= - 6. 45, P ＜ 0. 01).In subjective assessment, VNC images had more noise (2. 7 ± 0. 5 for VNC and 3.9 ± 0. 3 for CNC,Z = -6. 84, P ＜ 0. 01) and skull base-related artifacts (2. 4 ± 0. 9 for VNC and 3.7 ± 0. 5 for CNC,Z = -6. 15, P ＜0. 01) than CNC images. The gray/white matter contrast (1.3 ± 0. 5 for VNC and 3.3 ±0. 6 for CNC, Z = - 7. 01, P ＜ 0. 01), hyperdense lesion display (3.0 ± 0. 4 for VNC and 4. 0 ± 0. 0 for CNC,Z = -4. 52, P ＜ 0. 01) and hypodense lesion display (3.2 ± 0. 8 for VNC and 3.9 ± 0. 3 for CNC,Z= -3. 12, P ＜0. 01) on VNC images were lower than those on CNC images. In per-patient analysis,29 cases of hyperdense lesion (hemorrhage) were found on VNC images without misdiagnosis. The sensitivity, specificity, positive predictive value and negative predictive value were all 100. 0% (29/29,33/33, 29/29, 33/33). VNC images had the same detection rate of hyperdense lesions as CNC images (P ＞0. 05, Kappa = 1. 000) at per-patient level. Twenty-two patients with hypodense ischemic lesions were found on VNC images with one false positive case and two false negative cases. The sensitivity,specificity, positive predictive value and negative predictive value were 91.3% (21/23), 97.4%(38/39), 95.5% (21/22) and 95.0% (38/40) respectively. No statistical difference was found in detecting hypodense lesions between VNC and CNC images (χ2 = 0. 00, P ＞ 0. 05, Kappa = 0. 895). In per-lesion analysis, 53 hemorrhage lesions were found on VNC images with false negative results of four lesions and no false positive result. The sensitivity, specificity, positive predictive value and negative predictive value were 93.0% (53/57), 100. 0% (38/38), 100. 0% (53/53) and 90. 5% (38/42)respectively. There was no significant difference in detection rate of hyperdense lesion between VNC and CNC images (χ2 =2. 25, P ＞0. 05, Kappa =0. 914). Thirty-eight hypodense lesions were found on VNC images with 2 false positive lesions and 13 false negative lesions. The sensitivity, specificity, positive predictive value and negative predictive value were 73.5% (36/49), 96.4% (53/55), 94. 7% (36/38)and 80. 3% (53/66) respectively. The detection rate of hypodense lesion on VNC images was lower than that on CNC images (χ2 = 6. 67 ,P ＜ 0.01, Kappa = 0. 707). Conclusion Compared with CNC images,head VNC images have reduced image quality and radiation dosage. VNC images can replace CNC images potentially in detecting intracranial hemorrhage and provide information for ischemic cerebrovascular diseases to some extent.

OBJECTIVETo explore the protective effects of hirudin on acute experimental intracerebral hemorrhage (ICH) by observing the changes of histologic pathology and brain water content as well as GFAP-positive cells in the perihematomal brain regions.

METHODThe models of rat ICH were made with infusion of autologous blood into the right neucleus caudatus. The rats were divided randomly into control group, intracerebral hemorrhage group and treating group with hirudin. Brain water content was measured, and pathological and GFAP changes were observed.

RESULTThe pathological impairation after ICH were gradually deteriorated and peaked at the third day. Brain water content after ICH was gradually increased and obviously after one day(P < 0.05) and peaked at the third day. GFAP-positive cells were gradually increased and peaked at the seventh day after ICH. In the treating groups, the pathological impairation and brain water content as well as the GFAP-positive cells were decreased as compared to those in the intracerebral hemorrhage group and the control group. And the positive correlation between GFAP-positive cell numbers and brain water content were shown by linear regression.

CONCLUSIONThe local administration of hirudin, a special inhibitor of thrombin, has protective effects within the first week after ICH.

Acute Disease ; Animals ; Brain ; metabolism ; pathology ; Cerebral Hemorrhage ; metabolism ; pathology ; Female ; Glial Fibrillary Acidic Protein ; metabolism ; Hirudins ; pharmacology ; Male ; Neuroprotective Agents ; pharmacology ; Random Allocation ; Rats ; Rats, Wistar

AIM:To explore the regulatory effect of chemokine CCL 3 on exosome secretion from human bone marrow mesenchymal stem cells(hBMSCs).METHODS: hBMSCs were stimulated with chemokine CCL 3 at different concentrations in vitro.The proliferation of hBMSCs was measured by CCK-8 assay and viable cell counting.Exosome se-cretion from hBMSCs was qualitatively analyzed by transmission electron microscope(TEM)and flow cytometry, and the quantitative analysis was carried out by flow cytometry and nanoparticle tracking analysis(NTA).RESULTS:Compared with control group,the viability of the hBMSCs detected by CCK-8 assay was increased when hBMSCs were treated with CCL3(P<0.05).The results of viable cell counting demonstrated that the number of hBMSCs was raised in CCL 3 group in a dose-dependent manner(P<0.05).The results of flow cytometry showed that hBMSCs expressed 3 CCL3-related spe-cific receptors,CCR1,CCR5 and CCR9.Compared with control group,the fluorescence intensity of CCR9 in CCL3 group was obviously enhanced.However,no significant difference of fluorescence intensity for CCR 5 and CCR1 was observed be-tween the 2 groups.The results of NTA demonstrated that the secretion capacity of CCL 3-induced hBMSCs was far less than that in control group(P<0.05).However, the microvesicles larger than 100 nm in CCL3 groups were increased(P<0.05).The above results indicated that the higher concentration of CCL 3 induced the lower secretion of exosomes.In addi-tion,the results of flow cytometry demonstrated that CCL 3-induced hBMSCs showed lower quantity of CD 9 +exosomes than those in control group(P<0.01).CONCLUSION:CCL3 promotes the proliferation of hBMSCs but depresses the secre-tion of exosomes in a dose-dependent manner.CCL3 affects the size distribution of exosomes and increases the number of nonfunctional microvesicles of larger than 100 nm in size.CCL3 induces the expression of CCR9 in hBMSCs.

Objective To investigate the effects of dexmedetomidine preventive on chronic postsurgical pain (CPSP) in patients undergoing hysterectomy.Methods Eighty patients scheduled for elective abdominal hysterectomy,aged 18-65 years,ASA physical status Ⅰ or Ⅱ were recruited,and randomly divided into dexmedetomidine group (group D) and the control group (group C).All patients received total intravenous anesthesia with propofol and remifentanil.The patients in group D were administered intravenously dexmedetomidine 0.5μg·kg-1 ·h-1 from anesthesia induction to extubation at the end of surgery,while the patients in group C were administered normal saline 0.125 ml·kg-1· h-1.All patients received patient-controlled analgesia with fentanyl postoperatively.Intraoperative vital signs,the dose of analgesic and sedatives,and adverse reactions were recorded.CPSP and neuropathic pain (NP) were evaluated through the telephone follow-up in 3,6 and 12 months postoperatively.Results The peri-operative vital signs of both groups were stable,and no obvious adverse reaction were observed.The dosage of tramadol used for resue analgesia in group D was lower than that in group C [(58.8±15.4) mg vs (78.9±24.5) mg,P＜0.05].Seventy-one of eighty patients completed all follow-up (37 in group D,34 in group C).The incidence of CPSP in postoperative 3,6 and 12 months were 10.8％,5.4 ％,2.7 ％ in group D,significantly lower than 35.3 ％,26.5 ％,17.6％ in group C,respectively (P＜0.05).The incidence of NP in postoperative 3 and 6 months were 2.7％,0％,significantly lower than 17.6％,14.7％ in group C,respectively (P＜0.05).Conclusion Dexmedetomidine preventive analgesia alleviate chronic post-hysterectomy pain.