OBJECTIVETo investigate the ideal approach in creating rabbit model of hepatic fibrosis and to evaluate the feasibility and value of dynamic whole-liver 3D magnetic resonance (MR) perfusion-weighted imaging (PWI) in the quantitative study on the staging of hepatic fibrosis.

METHODSRabbit model of hepatic fibrosis was created by intraperitoneal injection of 5% and 100% carbon tetrachloride (0.1 ml/kg, once a week) respectively. MR perfusion weighted imaging was performed at the 6th, 8th, 10th and 12th week since injection. The time of peak (TOP), the time to peak (TTP), the maximum slope of increase(MSI) and the maximal relative signal increase (MRSI) of portal vein and hepatic parenchyma were analyzed quantitatively, and were compared with pathological results. Comparison of different concentrations of CCl4 was analyzed using chi-square test. Inter-group comparison of perfusion parameters was analyzed using one-way ANOVA P less than 0.05 was regarded as statistically significant.

RESULTS40% of the rabbits treated with 5% carbon tetrachloride developed hepatic fibrosis, while 75% of the rabbits treated with 100% carbon tetrachloride developed hepatic fibrosis; the mortality rate is significantly different between these two groups (X2=5.013, P less than 0.05). PWI examination was successfully achieved in 31 rabbits, liver perfusion baseline was stable, and good TIC curve was obtained. With the progress of hepatic fibrosis, TOP and TTP of portal vein and hepatic parenchyma were increased, and MSI and MRSI were decreased. There were significant differences among stage of S0-S2, S3 and S4.

CONCLUSIONSThe method (100% carbon tetrachloride intraperitoneal injection, 0.1 ml/kg, once a week) has high success rate of creating rabbit model of hepatic fibrosis. The stage of hepatic fibrosis could be evaluated quantitatively with dynamic whole-liver 3D MR perfusion-weighted imaging.

Animals ; Carbon Tetrachloride ; administration & dosage ; Disease Models, Animal ; Image Interpretation, Computer-Assisted ; methods ; Imaging, Three-Dimensional ; Liver ; blood supply ; diagnostic imaging ; pathology ; Liver Circulation ; Liver Cirrhosis, Experimental ; diagnosis ; diagnostic imaging ; Magnetic Resonance Angiography ; methods ; Male ; ROC Curve ; Rabbits ; Radiography ; Sensitivity and Specificity

BACKGROUNDCerebral hyperperfusion syndrome is an important complication of carotid endarterectomy (CEA). An >100% increase in middle cerebral artery velocity (MCAV) after CEA is used to predict the cerebral hyperperfusion syndrome (CHS) development, but the accuracy is limited. The increase in blood pressure (BP) after surgery is a risk factor of CHS, but no study uses it to predict CHS. This study was to create a more precise parameter for prediction of CHS by combined the increase of MCAV and BP after CEA.

METHODSSystolic MCAV measured by transcranial Doppler and systematic BP were recorded preoperatively; 30 min postoperatively. The new parameter velocity BP index (VBI) was calculated from the postoperative increase ratios of MCAV and BP. The prediction powers of VBI and the increase ratio of MCAV (velocity ratio [VR]) were compared for predicting CHS occurrence.

RESULTSTotally, 6/185 cases suffered CHS. The best-fit cut-off point of 2.0 for VBI was identified, which had 83.3% sensitivity, 98.3% specificity, 62.5% positive predictive value and 99.4% negative predictive value for CHS development. This result is significantly better than VR (33.3%, 97.2%, 28.6% and 97.8%). The area under the curve (AUC) of receiver operating characteristic: AUC(VBI) = 0.981, 95% confidence interval [CI] 0.949-0.995; AUC(VR) = 0.935, 95% CI 0.890-0.966, P = 0.02.

CONCLUSIONSThe new parameter VBI can more accurately predict patients at risk of CHS after CEA. This observation needs to be validated by larger studies.

Aged ; Blood Pressure ; physiology ; Cerebrovascular Circulation ; physiology ; Cerebrovascular Disorders ; physiopathology ; Endarterectomy, Carotid ; Female ; Hemodynamics ; physiology ; Humans ; Male ; Middle Aged ; Prospective Studies

OBJECTIVETo investigate the cell viabilities of vascular smooth muscle cells and vascular endothelial cells stimulated by cigarette smoke extract(CSE) .

METHODSThe CSE was prepared by smoke-bubbled phosphate buffered saline(PBS) generation.After culturing cells with different concentrations of CSE, we used the cell counting kit-8 to determine the cell viability.The expression levels of c-jun and cyclinD1 were analyzed through Western blot.The c-jun plasmid was transfected to detect the change of cyclinD1 expression.

RESULTSThe smooth muscle cell viability increased when the CSE concentration ranged 0.625%-10%, whereas the endothelial cells viability decreased when exposed to the CSE concentration. After exposure to CSE for 48 hours, there was no difference in c-jun expression between toxin group and PBS group;however, the expression of p-c-jun in the smooth muscle cells significantly increased in the toxin groups than in the PBS group(P<0.05) and the expression of p-c-jun in the vascular endothelial cells significantly decreased(P<0.05) . The level of cyclinD1 significantly increased after exposed to CSE, and its expression level also increased in respond to the c-jun overexpression.

CONCLUSIONCSE can enhance the proliferation of vascular smooth muscle cells and decrease in the activity of endothelial cells proliferation, which may be explained by the phosphorylation of c-jun and the expression of cyclinD1.

Cell Proliferation ; drug effects ; Cell Survival ; Cells, Cultured ; Cyclin D1 ; metabolism ; Endothelial Cells ; drug effects ; metabolism ; Humans ; Myocytes, Smooth Muscle ; drug effects ; metabolism ; Proto-Oncogene Proteins c-jun ; metabolism ; Tobacco ; adverse effects

OBJECTIVETo analyze the impact of efforts of community-based organizations (CBO) in HIV testing mobilization and case finding among men who have sex with men(MSM).

METHODSResults of HIV testing mobilization among MSM through CBOs in 15 program areas were collected and compared with corresponding HIV case reporting data to demonstrate the contribution of CBO-based HIV testing in HIV case finding among MSM from July 2008 to December 2011. Meanwhile,the proportion of screened HIV positives who received testing results notification,confirmatory test, following up and CD4 cell tests were analyzed and compared with those identified in medical institutions.

RESULTSA total of 196 075 HIV tests were performed for MSM, as a result of mobilization efforts of CBOs. Cumulatively 7704 new HIV cases were identified, accounting for 51.7% (7704/14 914) of all newly diagnosed HIV cases infected via homosexual sex in the program areas.Among the newly diagnosed MSM HIV infections in the program areas,the proportion of infections detected through the mobilization of CBOs increased from 35.4% (609/1722) in 2008 to 63.7% (2371/3722) in 2010, and 58.3% (3024/5189) in 2011. Compared with those identified through medical institutions, newly diagnosed MSM infections detected though CBOs testing mobilization have higher rates of receiving screening testing results notification (97.3% (4441/4563) vs 92.8% (13 140/14 153)) , (84.6% (2559/3024) vs 79.8% (5589/7002)) and CD4 cell tests (66.1% (1999/3024) vs 52.9% (3705/7002)), and a lower rate of receiving confirmatory test (78.6% (3588/4563) vs 85.6% (12 115/14 153)).

CONCLUSIONCBOs can take their advantages in mobilizing MSM to receive HIV test, and MSM HIV cases detected through CBOs have become the main source of MSM HIV case finding in program areas.

Community Health Services ; HIV Infections ; prevention & control ; HIV Seropositivity ; Health Promotion ; Homosexuality, Male ; Humans ; Male ; Mass Screening

BACKGROUNDOverexpression of G-protein coupled receptor 34 (GPR34) affects the progression and prognosis of human gastric adenocarcinoma, however, the role of GPR34 in gastric cancer development and progression has not been well-determined. The current study aimed to investigate the effect of GPR34 knockdown on the proliferation, migration, and apoptosis of HGC-27 gastric cancer cells and the underlying mechanisms.

METHODSThe expression of GPR34 in gastric cancer cell line HGC-27 was detected by quantitative real-time reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting. HGC-27 cells were employed to construct the stable GPR34 knockdown cell model in this study. Real-time RT-PCR and Western blotting were applied to validate the effect of short hairpin RNA (ShRNA) on the expression of GPR34 in HGC-27 gastric cells. The proliferation, migration of these cells were examined by Cell Counting Kit-8 and transwell. We also measured expression profile of PI3K/PDK1/AKT and ERK using Western blotting.

RESULTSThe ShRNA directed against GPR34 effectively inhibited both endogenous mRNA and protein expression levels of GPR34, and significantly down-regulated the expression of PIK3CB (P < 0.01), PIK3CD (P < 0.01), PDK1 (P < 0.01), phosphorylation of PDK1 (P < 0.01), Akt (P < 0.01), and ERK (P < 0.01). Furthermore, GPR34 knockdown resulted in an obvious reduction in HGC-27 cancer cell proliferation and migration activity (P < 0.01).

CONCLUSIONSGPR34 knockdown impairs the proliferation and migration of HGC-27 gastric cancer cells in vitro and provides a potential implication for therapy of gastric cancer.

Apoptosis ; genetics ; physiology ; Blotting, Western ; Cell Line, Tumor ; Cell Proliferation ; genetics ; physiology ; Humans ; RNA, Small Interfering ; genetics ; Real-Time Polymerase Chain Reaction ; Receptors, Lysophospholipid ; genetics ; metabolism ; Stomach Neoplasms ; genetics ; metabolism

OBJECTIVETo investigate the effect of interleukin-10 (IL-10) on the expression of transforming growth factor-beta(1) (TGFbeta(1)) and platelet-derived growth factor (PDGF) in hepatic stellate cells (HSC) during liver injury.

METHODSThe adenovirus vector (the titer was 1 x 10(7) efu/ml) encoded IL-10 gene was used to transfect the rat via the vein of caudal. At the same time, CCl(4) was injected into rat by a hypodermic injection. These processes went on twice a week. After eight weeks, the liver were perfused with collagenase IV and purified by density gradient centrifugation with Nycodenz for separate HSC. The level of IL-10 was measured by ELISA method; The expression of PDGF and TGFbeta(1) in HSC was detected by semi-quantitative RT-PCR and Western-blot methods.

RESULTSThe level of IL-10 in therapy group (adenovirus vector encoding IL-10 gene group) was higher than that in non-therapy group (adenovirus vector without IL-10 gene and PBS group); The expression of TGFbeta(1) mRNA, TGFbeta(1) protein and PDGF mRNA, PDGF protein in therapy group were significantly lower than that in non-therapy group (P < 0.05).

CONCLUSIONDownregulating the TGFbeta(1) and PDGF expression could be the passageway by which IL-10 alleviate the degree of proliferation and activation in hepatic stellate cells.

Animals ; Down-Regulation ; drug effects ; Genetic Therapy ; Hepatocytes ; drug effects ; physiology ; Interleukin-10 ; pharmacology ; Liver Cirrhosis, Experimental ; metabolism ; pathology ; therapy ; Male ; Platelet-Derived Growth Factor ; biosynthesis ; RNA, Messenger ; biosynthesis ; Rats ; Rats, Sprague-Dawley ; Stromal Cells ; drug effects ; physiology ; Transfection ; Transforming Growth Factor beta ; biosynthesis ; Transforming Growth Factor beta1

BACKGROUNDTransforming growth factor-beta1 (TGF-beta1) exerts strong fibrogenic potential in culture-activated HSCs. Smad4 is a key intracellular mediator for the transforming growth factor-beta (TGF-beta) superfamily of growth factors. The aim of this study was to assess the effects of the antisense Smad4 gene on Ito cell line, LI90.

METHODSThe recombinant retroviral vector pLXSN-Smad4 was constructed by cloning the rat antisense Smad4 cDNA into the retroviral vector pLXSN. Retroviruses with or without the antisense gene were obtained by transfecting pLXSN-Smad4 and pLXSN vectors into PA317 cells. Human hepatic stellate cells (HSCs) LI90 were infected with these retroviruses followed by selection with G418. The expression of Smad4 was detected by Northern and Western blots. Cell biological characteristics, including cell growth curve, 3H-TdR and 3H-proline uptake by HSCs and the production of extracellular matrix were assessed.

RESULTSmRNA and protein expressions of Smad4 in LI90 cells transfected with retrovirus containing the antisense Smad4 gene were much lower than those in LI90 cells transfected with empty vector or parental LI90 cells. Cells hypoexpressing the Smad4 gene exhibited a slower rate of growth, a lower uptake of 3H-TdR and 3H-proline (P < 0.01), and smaller production of th extracellular matrix, compared with parental LI90 cells and cells transfected with empty retrovirus.

CONCLUSIONSThe antisense Smad4 gene can suppress the expression of the Smad4 gene, reduce endogenous production of Smad4 mRNA and protein, block TGF-beta1 signaling pathway, inhibit activation of Ito cells, obstruct the growth of Ito cells, decrease the production of the extracellular matrix (ECM). Our results may provide a basis for the development of antifibrotic gene therapy.

Cell Line ; DNA, Antisense ; pharmacology ; DNA-Binding Proteins ; antagonists & inhibitors ; genetics ; Genetic Therapy ; Genetic Vectors ; genetics ; Humans ; Liver Cirrhosis ; therapy ; Retroviridae ; genetics ; Smad4 Protein ; Trans-Activators ; antagonists & inhibitors ; genetics ; Transfection ; Transforming Growth Factor beta ; physiology ; Transforming Growth Factor beta1

Animals ; Cells, Cultured ; Hepatocytes ; cytology ; metabolism ; Male ; Rats ; Rats, Sprague-Dawley ; Receptors, Somatostatin ; biosynthesis ; classification ; genetics

OBJECTIVETo evaluate the feasibility and safety of operative laparoscopy for ectopic pregnancy with hypovolemic shock.

METHODSTwo hundred and fifteen women with ectopic pregnancy underwent operative laparoscopy. These patients were divided into two groups. The study group included 21 patients with shock and intraperitoneal hemorrhage more than 1000 mL, and control group included 194 patients, hemodynamically stable, with blood loss less than 1000 mL. Clinical data of perioperative periods in two groups were retrospectively analyzed.

RESULTSAll patients were tubal pregnancies. The occurrence rate of tubal rupture was higher in study group than in control group (80.95% vs. 15.98%, P < 0.001). Intraabdominal blood loss was significantly higher in study group than in control group (1900 mL vs. 300 mL, P < 0.001), and autologous blood transfusions were given to 95.24% and 9.3% of patients in study and control group, respectively (P < 0.001). Laparoscopic salpingectomy was performed on 85.7 % and 50.5% of patients in study and control group (P < 0.001). The operative time was somewhat longer in study group than that in control group (60 minutes vs. 45 minutes), but with no significant difference. All patients had no perioperative complications.

CONCLUSIONOperative laparoscopy in patients with hopovolemic shock can be safely and effectively performed by experienced laparoscopists with the aid of optimal anesthesia, advanced cardiovascular monitoring, and autologous blood transfusion.

Adult ; Blood Loss, Surgical ; Blood Transfusion ; Fallopian Tubes ; surgery ; Female ; Gynecologic Surgical Procedures ; methods ; Humans ; Laparoscopy ; Pregnancy ; Pregnancy, Ectopic ; surgery ; Pregnancy, Tubal ; surgery ; Shock ; etiology ; surgery

OBJECTIVETo investigate the effect of somatostatin analogue-octreotide (OCT) on expression of connective tissue growth factor (CTGF) gene of murine hepatic stellate cells (HSCs) in vitro.

METHODSHSCs separated from Sprague Dawley rats by in situ perfusion and Nycodenz gradient were divided into 5 groups. HSCs in 4 out of 5 groups were co-cultured with octreotide at different dosages, and the remaining group served as control. The expression of CTGF and TGF-beta mRNA were assessed by reverse transcription-polymerase chain reaction (RT-PCR).

RESULTSOCT down-regulates the expression of CTGF and TGF-beta mRNA in HSCs. The effect is increased with a dose dependent manner.

CONCLUSIONSOCT could exert the inhibitory effect on HSCs by down-regulating the expression of CTGF and TGF-beta. This provides a potential for the prevention and management of hepatic fibrosis.

Animals ; Cells, Cultured ; Connective Tissue Growth Factor ; Gene Expression ; drug effects ; Hepatocytes ; drug effects ; metabolism ; Immediate-Early Proteins ; genetics ; Intercellular Signaling Peptides and Proteins ; genetics ; Male ; Octreotide ; pharmacology ; RNA, Messenger ; genetics ; Rats ; Rats, Sprague-Dawley ; Reverse Transcriptase Polymerase Chain Reaction ; Somatostatin ; analogs & derivatives ; Transforming Growth Factor beta ; genetics