الملخص
<p><b>BACKGROUND</b>Acute abdominal pain is a common symptom of emergency patients. The severity was always evaluated based on physicians' clinical experience. The aim of this study was to establish an early risk stratification method (ERSM) for addressing adults with acute abdominal pain, which would guide physicians to take appropriate and timely measures following the established health-care policies.</p><p><b>METHODS</b>In Cohort 1, the records of 490 patients with acute abdominal pain that developed within the past 72 h were enrolled in this study. Measurement data and numeration data were compared with analysis of variance and Chi-square test, respectively. Multiple regression analysis calculated odd ratio (OR) value. P and OR values showed the impacts of factors. ERSM was established by clinical experts and statistical experts according to Youden index. In Cohort 2, data from 305 patients with acute abdominal pain were enrolled to validate the accuracy of the ERSM. Then, ERSM was prospectively used in clinical practice.</p><p><b>RESULTS</b>The ERSM was established based on the scores of the patient's clinical characteristics: right lower abdominal pain + 3 × diffuse abdominal pain + 3 × cutting abdominal pain + 3 × pain frequency + 3 × pain duration + fever + 2 × vomiting + 5 × stop defecation + 3 × history of abdominal surgery + hypertension history + diabetes history + hyperlipidemia history + pulse + 2 × skin yellowing + 2 × sclera yellowing + 2 × double lung rale + 10 × unconsciousness + 2 × right lower abdominal tenderness + 5 × diffuse abdominal tenderness + 4 × peritoneal irritation + 4 × bowel sounds abnormal + 10 × suspicious diagnosis + white blood cell count + hematocrit + glucose + 2 × blood urea nitrogen + 3 × creatine + 4 × serum albumin + alanine aminotransferase + total bilirubin + 3 × conjugated bilirubin + amylase. When the score was <18, the patient did not need hospitalization. A score of ≥18 and <38 indicated that the patient should be under observation or hospitalized. A score of ≥38 and <50 indicated the need for an emergent operation. A score of ≥50 indicated the need for admission to the Intensive Care Unit. The area under the receiver operating characteristic curve of the ERSM in Cohorts 1 and 2 were 0.979 and 0.988, respectively.</p><p><b>CONCLUSIONS</b>This ERSM was an accurate and reliable method for making an early determination of the severity of acute abdominal pain. There was the strong correlation between scores of ERSM and health-care decision-making.</p>
الموضوعات
Adult , Aged , Aged, 80 and over , Female , Humans , Male , Middle Aged , Young Adult , Abdominal Pain , Diagnosis , Chi-Square Distribution , Cohort Studies , Intensive Care Units , ROC Curve , Risk Assessment , Methods , Risk Factorsالملخص
<p><b>OBJECTIVE</b>To isolate antifungal compound from Paeonia suffruticosa, and to find the antifungal mechanisms by observing the ultrastructural modifications of yeasts in growth phase produced by 1,2,3,4,6-penta-O-galloyl-beta-D-glucose (PGG).</p><p><b>METHODS</b>Peony (Paeonia suffruticosa) root bark (PRB) was separated by solvent extraction and purified by high performance liquid chromatography (HPLC) method using analytical and preparative reversed phase C18 column on the basis of bio-assay method. In order to investigate the antifungal mechanism of PGG, Yeasts were submitted to different concentrations [3 × minimum inhibition concentration (MIC), 0.3 × MIC] for 1 h under constant stirring at 30 °C, and transmission electron microscopy was performed.</p><p><b>RESULTS</b>Based on the antifungal activity of PRB on Candida glabrata CBS138, the antifungal compound were isolated in ethyl acetate layer of PRB and identified as PGG by mass spectrometry, 1H nuclear magnetic resonance (NMR) analyses, with molecular weight of 940 and molecular formular as C41H32O26. Transmission electron microscopy showed that PGG degraded the cell wall envelope.</p><p><b>CONCLUSION</b>The results suggest that PGG may be responsible for the antifungal activity of PRB by disrupting the structure of cell wall directly.</p>
الموضوعات
Antifungal Agents , Chemistry , Pharmacology , Candida , Chromatography, High Pressure Liquid , Hydrolyzable Tannins , Chemistry , Pharmacology , Mass Spectrometry , Microbial Sensitivity Tests , Paeonia , Chemistry , Plant Bark , Chemistry , Plant Extracts , Pharmacology , Plant Roots , Chemistry , Proton Magnetic Resonance Spectroscopyالملخص
<p><b>OBJECTIVE</b>To investigate the effects of Herbal Compound 861 (Cpd 861) on collagen synthesis and degradation in rat mesangial cells exposed to high glucose.</p><p><b>METHODS</b>The third to fifth passage of rat mesangial cells were exposed to high glucose and Cpd 861 at a concentration of 0.25-4.00 g/L for 24, 48 and 72 h, respectively. Benazepril (10(-7)-10(-3) mmol/L) was selected as positive control. The methyl thiazolyl tetrazolium colorimetric assay was used to evaluate the effect of Cpd 861 on cell proliferation. After incubation with Cpd 861 at a concentration of 2.00 g/L for 48 h, the protein secretions of collagen type IV, matrix metallopeptidase 9 (MMP-9), tissue inhibitor of metalloproteinase-1 (TIMP-1), transforming growth factor beta 1 (TGF-β1), and hepatocyte growth factor (HGF) were detected by enzyme-linked immunosorbent assay method. And rat mesangial cells were harvested to determine MMP-9, TIMP-1, TGF-β1 and HGF mRNA expression by reverse transcription polymerase chain reaction.</p><p><b>RESULTS</b>Cpd 861 inhibited cell proliferation induced by high glucose in a dose- and time-dependent manner. Compared with high glucose, collagen type IV production was decreased significantly by Cpd 861 (P<0.01). Cpd 861 increased the protein secretions and mRNA expressions of MMP-9 and HGF, whereas the protein secretions and mRNA expressions of TIMP-1 and TGF-β1 were reduced markedly (P<0.05). The ratio of MMP-9 to TIMP-1 was enhanced by Cpd 861 significantly. There was no significant difference in all above-mentioned effects between Cpd 861 (2.00 g/L) and benazepril (10(-5) mmol/L).</p><p><b>CONCLUSION</b>The anti-glomerulosclerosis mechanisms of Cpd 861 were partly attributed to its effects of inhibiting mesangial cell proliferation, decreasing collagen synthesis and enhancing collagen degradation.</p>
الموضوعات
Animals , Rats , Cell Proliferation , Cells, Cultured , Collagen Type IV , Bodily Secretions , Drugs, Chinese Herbal , Pharmacology , Fibrosis , Glucose , Toxicity , Hepatocyte Growth Factor , Bodily Secretions , Matrix Metalloproteinase 9 , Metabolism , Mesangial Cells , Cell Biology , Metabolism , Polymerase Chain Reaction , Proteolysis , RNA, Messenger , Genetics , Metabolism , Tissue Inhibitor of Metalloproteinase-1 , Metabolism , Transforming Growth Factor beta1 , Bodily Secretionsالملخص
To analyze the characteristics of serum sodium in decompensated cirrhosis and evaluate the prognostic ability of the model for end-stage liver disease (MELD) in Na-containing models. Patients diagnosed with decompensated cirrhosis at our hospital were enrolled for study between June 2005 and October 2010. Patients were classified among three groups, according to serum sodium concentration: less than 125 mmol/L, 125 to 135 mmol/L, and more than 135 mmol/L. Mortality rates among the three groups were compared by Kaplan-Meier survival analysis. In addition, the different serum sodium concentrations were analyzed for correlations between Child-Pugh score and complication incidence rates of portal hypertension. The area under the receiver operating characteristic (ROC) curve (AUC) was used to compare the predictive abilities of MELD, MELD-Na, and the integrated (i) MELD scores for 3-month, 6-month and 1-year mortalities. A total of 467 patients were analyzed, and 50.54% had hyponatremia ( less than 135 mmol/L). Sodium concentration was correlated with mortality, and Kaplan-Meier survival analysis indicated that mortality was significantly higher in each subgroup with lower sodium concentration (all, P = 0.000). Likewise, sodium concentration decreased in conjunction with increased severity of decompensation, as classified by Child-Pugh scoring (sodium: A more than B more than C; mortality: A less than B less than C). With the exception of digestive tract bleeding, complication incidence rates of hepatic encephalopathy, ascites, spontaneous bacterial peritonitis, and hepatorenal syndrome increased when sodium concentration decreased. For predicting 3-month mortality, the AUC scores of MELD were not significantly different from the MELD-Na and iMELD scores (P more than 0.05). For predicting 6-month and 1-year mortality, the AUC scores of MELD-Na and iMELD were significantly higher than those of MELD (P less than 0.05). Hyponatremia is correlated with mortality and complications in decompensated cirrhosis patients. Incorporation of Na into the MELD may enhance it's prognostic ability.
الموضوعات
Adult , Aged , Female , Humans , Male , Middle Aged , End Stage Liver Disease , Liver Cirrhosis , Blood , Liver Failure , Diagnosis , Predictive Value of Tests , Prognosis , Serum , Chemistry , Severity of Illness Index , Sodium , Bloodالملخص
<p><b>OBJECTIVE</b>To determine the relationship of thick greasy tongue fur formation and permeability of vascular endothelial cells (ECs) with the protein expression of zonula occludens-1 (ZO-1).</p><p><b>METHODS</b>Sprague Dawley rats were randomly divided into a model group of severe acute pancreatitis (SAP) and a sham-operated (SO) group. The SAP rats were further divided into two subgroups on the basis of tongue-coating status: a thick greasy tongue fur group (SAP-TGF) and a normal tongue fur group (SAP-NF). Six rats were chosen randomly from every group mentioned above for an Evans blue assay 5 days after model establishment. For the histomorphology analysis, the expressions of ZO-1 protein and mRNA were studied by hematoxylin-eosin (H&E) staining, transmission electron microscope, Western blot, and Q-PCR using blood and tongue tissues, which were collected from 8 rats randomly chosen from each group.</p><p><b>RESULTS</b>The papillae density of the rat tongue surface and the caryocinesis frequency of the basal layer were significantly increased in the SAP-TGF group compared with the SO group (P<0.05). Evans blue levels in the tongue tissue of the SAP-TGF group were significantly higher than that of the SO and SAP-NF groups (P<0.05). Vascular ECs were wider and obviously fissured in the SAP-TGF group under transmission electron microscope observation. The protein and mRNA expression of ZO-1 in the SAP-TGF group were lower than those in the SAP-NF (P<0.05).</p><p><b>CONCLUSIONS</b>Reproductive activity enhancement of glossal epithelial cells was one of the main characteristics of thick greasy tongue fur formation. An increase in vasopermeability was closely associated with thick greasy tongue fur formation. Tight junction structural variation of vascular ECs might play an important role in the pathological and physiological process of thick greasy tongue fur formation.</p>
الموضوعات
Animals , Male , Rats , Blotting, Western , Cell Membrane Permeability , Endothelial Cells , Cell Biology , Evans Blue , Metabolism , Gene Expression Regulation , Membrane Proteins , Genetics , Metabolism , Phosphoproteins , Genetics , Metabolism , RNA, Messenger , Genetics , Metabolism , Rats, Sprague-Dawley , Tongue , Pathology , Zonula Occludens-1 Proteinالملخص
<p><b>BACKGROUND</b>The infection rate of methicillin-resistant Staphylococcus aureus (MRSA) is increasing yearly due to the overprescription of antibiotics. Traditional Chinese compound medicines are less inclined to induce bacterial resistance in the clinical setting because of their multi-acting mechanisms. However, most current research is limited to bacteriostasis in vitro using single extracts or formulations. Plasma pharmacology is an in vitro method, using what is called "medicine serum". The aim of this study was to investigate whether the medicine serum of compound Qingre granules (QRKL) alone or in combination with antibiotics may treat MRSA infection in the clinic.</p><p><b>METHODS</b>An animal model of MRSA resistance was created by injecting rabbits with the standard strain of MRSA ATCC43300. Infected rabbits were treated with QRKL by intragastric administration. Sixty minutes after the last intragastric administration, serum was obtained from the rabbits by heart puncture to obtain what is termed "medicine serum". The minimum inhibitory concentration (MIC) of QRKL, medicine serum alone, or serum combined with antibiotics was assessed by agar dilution.</p><p><b>RESULTS</b>were compared with the growth of sixteen isolates of MRSA.</p><p><b>RESULTS</b>The MIC of QRKL to the standard strain ATCC43300 was 10.00 mg/ml. The MIC(90) of vancomycin was 1.00 microg/ml, which, when combined with QRKL, dropped to 0.50 microg/ml. The MIC(90) of cefuroxime alone was 512.00 microg/ml. This level also decreased to 256.00 microg/ml when combined with QRKL. The addition of QRKL thus significantly reduced the MIC of both cefuroxime and vancomycin compared with antibiotics alone (P < 0.01). The MIC(90) of vancomycin with medicine serum decreased to 0.50 microg/ml, and the MIC of vancomycin with medicine serum also descended compared with using vancomycin alone (P < 0.01).</p><p><b>CONCLUSIONS</b>The growth of MRSA can be inhibited by QRKL or medicine serum of QRKL in vitro. The addition of QRKL results in increased sensitivity of MRSA to vancomycin and this may provide a novel treatment for patients with MRSA infection.</p>
الموضوعات
Drugs, Chinese Herbal , Pharmacology , Methicillin-Resistant Staphylococcus aureus , Microbial Sensitivity Tests , Vancomycin , Pharmacologyالملخص
<p><b>OBJECTIVE</b>To investigate the effects of transforming growth factor beta (TGF ) on c-fos gene expression in hepatic stellate cells.</p><p><b>METHODS</b>Hepatic stellate cells (HSC-T6) were cultured in the medium containing different concentrations of TGF (0.2, 1, and 5 ng/ml), and cells were collected at different time points of incubation (8, 24, 48, and 72 h). The total RNA of the HSCs was isolated and c-fos gene expression level were measured by reverse transcription polymerase chain reaction.</p><p><b>RESULTS</b>c-fos gene expression levels of HSCs cultured in the presence of low (0.2 ng/ml), moderate (1 ng/ml) and high (5 ng/ml) concentrations of TGF for 8, 24, 48 and 72 h were significantly greater than those of control group. The c-fos gene expression levels of HSCs increased gradually with the increment of TGF concentration, and significant differences in c-fos gene expression were found between the 3TGF groups.</p><p><b>CONCLUSION</b>TGF strongly up-regulates c-fos gene expression in hepatic stellate cells.</p>
الموضوعات
Animals , Rats , Cells, Cultured , Gene Expression , Genes, fos , Hepatic Stellate Cells , Proto-Oncogene Proteins c-fos , Genetics , Metabolism , Transforming Growth Factor beta , Pharmacologyالملخص
<p><b>OBJECTIVE</b>To develop diagnostic criteria of multiple organ dysfunction syndrome (MODS) by a prospective and multi-center clinical investigation.</p><p><b>METHODS</b>The data of 1087 MODS cases obtained from ICU of 37 hospitals from March 2002 to January 2005 in 11 provinces in China were analyzed in order to derive the diagnostic criteria of MODS.</p><p><b>RESULTS</b>This MODS diagnostic criteria involved 7 organs. To diagnose MODS, the original cause of MODS should be identified, then there should be two or more organs showing signs of dysfunction. The criteria for organ dysfunction were as follows. (1) Cardiovascular system: SBP < 90 mm Hg (1 mm Hg = 0.133 kPa), MAP < 70 mm Hg, signs of shock, ventricular tachycardia, ventricular fibrillation, or myocardial infarction; (2) Respiratory system: oxygenation index < 300 mm Hg; (3) Nervous system: indifference, restlessness, lethargy, light coma, or deep coma, Glasgow score < or = 14; (4) Blood system: PLT < 100 x 10(9)/L; CT, APTT, and PT prolonged or shortened; positive plasma protamine paracoagulation; (5) Liver: TBIL > 20. 5 micromol/L, ALB < 28 g/L; (6) Kidney: Cr > 123.8 micromol/L, urinary volume < 500 ml/24 h; (7) Gastro-intestine: bowel sounds decreased or disappeared; retention in the stomach, or positive occult blood feces with dark stools or haematemesis; intraabdominal pressure (intravesical pressure) > or = 11 cm H2O (1 cm H2O = 0.098 kPa). Any organ function met with one of the above conditions was considered to have dysfunction.</p><p><b>CONCLUSIONS</b>This diagnostic criterion of multiple organ dysfunction syndrome has been developed by this research, but it needs to accumulate experience by clinical practice and to revise the diagnosis criteria.</p>
الموضوعات
Adolescent , Adult , Aged , Aged, 80 and over , Female , Humans , Male , Middle Aged , Young Adult , Multiple Organ Failure , Diagnosis , Prospective Studiesالملخص
<p><b>BACKGROUND</b>Hepatitis B is at particularly high risk of fibrosis progression. Unfortunately, the mechanism of hepatic fibrogenesis induced by hepatitis B virus (HBV) has not been fully understood to date. The aim of this study was to observe whether HBV can infect hepatic stellate cells (HSCs), and to examine the effects of HBV or HBV S protein (HBs) on the proliferation and collagen type I expression of HSCs.</p><p><b>METHODS</b>The supernatants of HepG2.2.15 cells which contained HBV-DNA or HBs were added to LX-2 cells for 72 hours. Cell survival was determined by MTT assay. HBV particles in LX-2 cells were detected by transmission electron microscopy. The expression of HBs and HBV C protein (HBc) was determined by confocal fluorescence microscopy. The expression levels of HBV-DNA were measured by real-time PCR. The cellular collagen type I mRNA and protein levels were quantified by reverse transcription-PCR and ELISA, respectively.</p><p><b>RESULTS</b>High concentrations of HBV (1.2 x 10(5) - 5.0 x 10(5) copies/ml) or HBs (1.25 - 20 microg/ml) inhibited the proliferation of LX-2 cells, while low concentrations of HBV (1.0 x 10(3) - 6.2 x 10(4) copies/ml) or HBs (0.04 - 0.62 microg/ml) promoted the proliferation. After treating LX-2 cells with HBV for 72 hours, about 42 nm HBV-sized particles and strong expression of HBs and HBc were found in the cytoplasm of LX-2 cells. HBV-DNA in the culture medium of LX-2 cells decreased at 24 hours, rose at 48 hours and thereafter, decreased again at 72 hours. The mRNA and protein expression of cellular collagen type I in LX-2 cells were significantly increased by HBV infection but not by recombinant HBs.</p><p><b>CONCLUSIONS</b>HBV and HBs affect the proliferation of HSCs; HBV can transiently infect and replicate in cultured HSCs and express HBs and HBc in vitro. Furthermore, HBV can significantly increase the expression of collagen type I mRNA and protein in HSCs.</p>
الموضوعات
Humans , Cell Line , Cell Proliferation , Collagen Type I , Metabolism , Enzyme-Linked Immunosorbent Assay , Gene Expression Regulation , Hepatic Stellate Cells , Metabolism , Virology , Hepatitis B virus , Physiology , Microscopy, Confocal , Microscopy, Electron, Transmission , Polymerase Chain Reaction , Reverse Transcriptase Polymerase Chain Reactionالملخص
<p><b>OBJECTIVE</b>To evaluate the effect of portal hypertension on prognosis in patients with decompensated liver cirrhosis.</p><p><b>METHODS</b>The clinical data of decompensated cirrhosis patients in our hospital, between 2003 and 2006, were retrospected and followed up. Model for end-stage liver disease (MELD) score and Child-Turcotte-Pugh (CTP) classification was calculated using the standard formula. Kaplan-Meier survival analysis was used to compare the mortality in subgroups ranked by the syndromes. Cox proportional hazards regression was used to evaluate the effect of the syndromes on prognosis.</p><p><b>RESULTS</b>A cohort of 322 patients was admitted in this study at the end of the follow-up. The mortality of variceal bleeding, hepatic encephalopathy, a large volume ascites, spontaneous bacterial peritonitis, the type I and type II hepatorenal syndrome was 45.9%, 79.4%, 66.7%, 100%, 100% and 84.6% respectively. On the whole, the occurrence of all the syndromes was correlated with CTP classification and MELD score. Kaplan-Meier survival analysis showed that all of these syndromes, except for low to medium volume of ascites, significantly affected the survival rate (P<0.01). In Cox regression analysis, all the syndromes were the independent predictors of prognosis, the regression coefficient values of hepatic encephalopathy, spontaneous bacterial peritonitis, type I and type II hepatorenal syndrome, variceal bleeding and ascites were 0.973, 0.928, 0.935, 0.866, 0.464 and 0.369 respectively.</p><p><b>CONCLUSIONS</b>The portal hypertensive syndromes have significant effect on the prognosis of the patients with decompensated cirrhosis, hepatic encephalopathy is the worst one.</p>
الموضوعات
Adult , Aged , Female , Humans , Male , Middle Aged , Esophageal and Gastric Varices , Epidemiology , Follow-Up Studies , Gastrointestinal Hemorrhage , Epidemiology , Hepatic Encephalopathy , Epidemiology , Hepatorenal Syndrome , Epidemiology , Hypertension, Portal , Epidemiology , Liver Cirrhosis , Mortality , Pathology , Models, Statistical , Predictive Value of Tests , Prognosis , Severity of Illness Index , Survival Analysisالملخص
<p><b>OBJECTIVE</b>To explore the relationship of ascitic bacterial DNA and plasma endotoxin, intestinal permeability, gut flora in cirrhotic patients.</p><p><b>METHODS</b>Fifty-five decompensated cirrhotic patients with ascites were included in the study. A paracentesis was performed for every patient at admission and ascites fluid was collected for bacterial DNA isolation and amplification, plus other routine studies and cultured for aerobic and anaerobic bacteria. Plasma endotoxin, intestinal permeability, and gut flora were studied on the day following admission. Blood from the patients was obtained for routine hematologic, biochemical, and coagulation studies. Thirty healthy subjects were studied as normal controls.</p><p><b>RESULTS</b>No bacteria were found in the ascites cultures in the 55 patients, but bacterial DNA was found in 19 (34.55%). Compared with the bacterial DNA negative group, the bacterial DNA positive group showed a significantly lower level of PTA (t= -3.184, P=0.002), a higher Child-Pugh score (t=3.224, P=0.002) and a higher quantity of WBC in their ascitic fluid (t=4.088, P=0.001). Compared with normal controls, cirrhotic patients showed significantly higher levels of plasma endotoxin (t=13.705, P=0.000), lactulose/mannitol (L/M, t=28.568, P=0.000) in urine, and the quantity of enteric bacilli (t=2.912, P=0.005); the quantity of their intestinal bifidobacteria was significantly lower (t= -3.669, P=0.000). The variables correlative with the presence of bacterial DNA were the quantities of enteric bacilli (P=0.007) and PTA (P=0.011).</p><p><b>CONCLUSION</b>Intestinal bacterial overgrowth plays a key role in the pathogenesis of ascitic bacterial translocation (ABT) in cirrhotic patients. ABT is correlated with the degrees of the liver disease.</p>
الموضوعات
Adult , Aged , Aged, 80 and over , Female , Humans , Male , Middle Aged , Ascites , Microbiology , Ascitic Fluid , Microbiology , Bacterial Translocation , DNA, Bacterial , Enterobacteriaceae , Genetics , Liver Cirrhosis , Microbiologyالملخص
<p><b>OBJECTIVE</b>To explore the intestinal mucosal barrier protective effect of herbal medicine Compound Tongfu Granule (CTG) in patients with liver cirrhosis of decompensation stage.</p><p><b>METHODS</b>Fifty patients enrolled were randomly assigned to the control group (26 cases) and the CTG group (24 cases), and 30 healthy adults were set up as normal control. After 2-week treatment, the intestinal permeability (IP, represented by urinary lactulose/mannitol excretion rate), plasma endotoxin (EDT) level, and change of enteric bacteria (EB) in patients were observed before and after treatment, and compared with those in the normal control.</p><p><b>RESULTS</b>Before treatment, cirrhotic patients showed significantly higher levels of IP, EDT, and intestinal bacilli, but a lower amount of enteric bifidobacteria as compared with those the normal control. After 2-week treatment, levels of EDT and urinary excretion rate of lactulose in the CTG group were lowered more significantly than those in the control group (P < 0.05), while the amount of bifidobacteria in the CTG group increased accompanied with intestinal bacilli significantly lowered to near the levels in the normal control (P < 0.05, P < 0.01).</p><p><b>CONCLUSION</b>CTG can improve the intestinal barrier function, correct the intestinal bacteria disturbance, and significantly reduce the entero-derived endotoxemia in cirrhotic patients of decompensation stage.</p>
الموضوعات
Adult , Aged , Aged, 80 and over , Humans , Male , Middle Aged , Bifidobacterium , Metabolism , Cell Membrane Permeability , Drugs, Chinese Herbal , Therapeutic Uses , Endotoxins , Metabolism , Intestinal Mucosa , Metabolism , Microbiology , Pathology , Lactulose , Metabolism , Liver Cirrhosis , Drug Therapy , Metabolism , Microbiology , Pathology , Treatment Outcomeالملخص
<p><b>OBJECTIVE</b>To test the efficiency of infection of recombinant adeno-associated virus (rAAV) carrying hepatitis B virus S, C or X antigen, rAAV-HBV-S, C, X to human dendritic cells.</p><p><b>METHODS</b>Recombinant AAV plasmids containing HBV-S, C or X gene were constructed and packaged into rAAV in 293 cells. Monocytes were isolated from healthy donor and pulsed by rAAV-HBV-S, X, C or 293 lysate as control at the first day of isolation, then the dentritic cells were cultured for 7 days in vitro. The transcription and expression of HBV-S, C or X gene were analyzed by reverse transcription-polymerase chain reaction (RT-PCR) or intracellular staining with fluorescence activated cell sorter (FACS), respectively.</p><p><b>RESULTS</b>The titer of rAAV-HBV-S, C, X virus was approximately 10(-7) copies per ml. After infection the HBV-S, C or X transcription expression could be seen by RT-PCR in the infected dendritic cells, the efficiency was about 90 percent by FACS.</p><p><b>CONCLUSION</b>rAAV-HBV-c can effectively infect and pulse dendritic cells.</p>
الموضوعات
Humans , Cell Line , Cells, Cultured , Dendritic Cells , Cell Biology , Virology , Dependovirus , Genetics , Metabolism , Gene Expression , Genetic Vectors , Genetics , Metabolism , Hepatitis B Surface Antigens , Genetics , Metabolism , Transduction, Genetic , Methodsالملخص
<p><b>OBJECTIVE</b>To investigate the anti-fibrotic effects of Qishen Yiqi Dropping Pills (QYDP) in rats with liver fibrosis (LF).</p><p><b>METHODS</b>The LF model was induced by intraperitoneal injection with dimethylnitrosamine (DMN). Sixty Wistar rats were randomly divided into the normal group, the model group A, the QYDP intervened group , the model group B , and the QYDP treated group B. The degree of LF was evaluated according to 6-phase grading method. The expressions of collagen type I and III and tissue inhibitor of metalloproteinase-1 (TIMP-1) in liver tissues were determined by immunohistochemistry and the levels of collagen type I and III and TIMP-1 mRNA determined by semi-quantitive RT-PCR.</p><p><b>RESULTS</b>Compared with the model group A and B, the degree of LF, the positive expressions of TIMP-1 mRNA and the content of collagen type I and III in liver tissue in the QYDP intervened and treated groups were significantly lower.</p><p><b>CONCLUSION</b>QYDP could reduce the pathological changes and degree of LF in rats, which may be partially through inhibiting the expressions of collagen type I and III and TIMP-1.</p>
الموضوعات
Animals , Male , Rats , Collagen Type I , Genetics , Collagen Type III , Genetics , Drugs, Chinese Herbal , Therapeutic Uses , Immunohistochemistry , Liver Cirrhosis, Experimental , Drug Therapy , Phytotherapy , RNA, Messenger , Genetics , Random Allocation , Rats, Wistar , Reverse Transcriptase Polymerase Chain Reaction , Tissue Inhibitor of Metalloproteinase-1 , Geneticsالملخص
<p><b>OBJECTIVE</b>To compare the complication rates induced by three liver biopsy methods.</p><p><b>METHODS</b>One thousand five hundred fifty-seven liver biopsies were performed from 1995 to 2006 by the senior author of the present article. The patients were grouped into a non-ultrasound-guided biopsy group (783 cases), a partially ultrasound-guided group (485 cases) and an ultrasound-guided group (289 cases). The observed complications such as death, hemorrhage, pneumothorax, pain, choleperitoneum and vasovagal syncope were compared between these three groups.</p><p><b>RESULTS</b>Only 1 patient in the non-ultrasound-guided group died. Seven patients (45%), 4 in non-ultrasound-guided group, 2 in partially ultrasound-guided group and 1 in ultrasound-guided group, had hemorrhage complications. The hemorrhage complication rates between partially ultrasound-guided group and ultrasound-guided group had no significant differences and both were lower than those of the non-ultrasound-guided group. Thirty-five patients, 23 in non-ultrasound-guided group and 12 in partially ultrasound-guided group, suffered from pneumothorax; but the rates between the two groups were not significantly different. Forty-eight patients (45%), 28 in non-ultrasound-guided group, 17 in partially ultrasound-guided group and 3 in ultrasound-guided group, had severe pain and the rate in the ultrasound-guided group was significantly lower than those of the other two groups.</p><p><b>CONCLUSION</b>Liver biopsy performed under ultrasound guidance is the safest and most reliable way to do the procedure. It should be recommended in clinical practices.</p>
الموضوعات
Adolescent , Adult , Aged , Child , Humans , Middle Aged , Young Adult , Biopsy , Methods , Liver , Diagnostic Imaging , Pathology , Ultrasonographyالملخص
<p><b>OBJECTIVES</b>To analyze the frequency and the clinical and virological features of HBeAg-negative and HBeAg-positive chronic hepatitis B.</p><p><b>METHODS</b>Four hundred and seventeen chronic hepatitis B patients, 286 males and 131 females seen in our center were studied. Liver biopsies were taken from 83 patients.</p><p><b>RESULTS</b>The cases with HBeAg-negative chronic hepatitis B were 241 (57.8%), with an average age of 43.7+/-10.8 and a history of 16.8+/-8.5 years. HBeAg-positive chronic hepatitis B cases were 176 (42.2%), with an average age of 36.95+/-11 and a history of 12.3+/-8.0 years. HBeAg-negative patients were significantly older (P < 0.01) in age and had a longer disease history. ALT levels and the percentage of HBV DNA were higher than 10(5) copies/ml in HBeAg-negative patients and were significantly lower than those in the HBeAg-positive patients [(37.66+/-32.93) U/L vs. (82.09+/-107.57) U/L, 38.2% vs. 94.3%, P < 0.01]. Liver biopsies from 47 HBeAg-negative patients showed that the number of cases with inflammation scores of G1, G2, G3 and G4 were 5, 27, 14, 1 and the number of cases with fibrosis scores of S1, S2, S3 and S4 were 10, 12, 5, 20, respectively. In the 36 HBeAg-negative patients the respective number of cases with inflammation scores of G1, G2, G3 and G4 were 5, 14, 15, 2, and with fibrosis scores of S1, S2, S3, S4 were 8, 12, 6, 10. Although histopathological inflammation and fibrosis scores had no statistical difference between HBeAg-negative and positive patients (P > 0.05), 53.2% patients of HBeAg-negative group and 44.5% patients of HBeAg-positive group had a fibrosis score of >or= S3.</p><p><b>CONCLUSION</b>Despite lower serum ALT and HBV DNA, HBeAg-negative chronic hepatitis B still has a significant disease progression. This observation may help to develop better clinical management in HBeAg-negative chronic hepatitis B patients.</p>
الموضوعات
Adult , Female , Humans , Male , Middle Aged , Hepatitis B e Antigens , Blood , Hepatitis B, Chronic , Blood , Pathology , Liver , Pathologyالملخص
<p><b>OBJECTIVE</b>To develop a scoring system to measure the severity of the multiple organ dysfunction syndrome (MODS).</p><p><b>METHODS</b>The clinical data of patients with MODS were collected and analyzed prospectively. Seven indicators were screened out to assess the functions of seven organs. Each indicator was scored from 0 to 4 points, with the 0 point representing the normal organ function, and 1 point to 4 points representing the dysfuntion to failure of the organ. Acording to the index and points a severity scoring system were developed.</p><p><b>RESULTS</b>Seven indicators including the systolic pressure, oxygenation index, conscious state, peripheral blood platelet count, blood total bilirubin, and serum creatinine were used to represent the functions of seven organs. The seven organs included cardiovascular system, lung, brain, coagulative system, liver, kidney, and gastrointestinal tract. The severity scoring system were composed of seven indicators and their points. The total score was 24 points. The mortality increased along with the increase of the points (P<0.001).</p><p><b>CONCLUSION</b>This scoring system can be used to assess the severity of the MODS.</p>
الموضوعات
Humans , Multiple Organ Failure , Diagnosis , Mortality , Severity of Illness Indexالملخص
<p><b>OBJECTIVES</b>Elevated tissue inhibitor of metalloproteinase-1 (TIMP-1) expression contributes to excess extracellular matrix in liver fibrosis. This study was designed to construct two recombinant adeno-associated viruses (AAV) carrying antisense RNA and small interfering RNA (siRNA) of TIMP-1 (rAAV/ANTI-TIMP-1/neo and rAAV/siRNA-TIMP-1/neo), and then to compare the suppression of TIMP-1 gene expression on rat hepatic stellate cell (HSC)-T6 cells infected by these two types of viruses in vitro.</p><p><b>METHODS</b>Antisense RNA amplified by rat HSC-T6 and U6 promoter followed by the annealing siRNA were cloned into the AAV vector (pdl6-95/neo) and packed in 293 cells to construct the recombinants rAAV/ANTI-TIMP-1/neo and rAAV/siRNA-TIMP-1/neo. Rat HSC-T6 cells were infected with these recombinant AAVs and selected by using G418, and real-time PCR after reverse transcription and Western blot were performed to detect the transcription and expression level of TIMP-1 gene in these cells.</p><p><b>RESULTS</b>The results of PCR, restrictive enzyme digestion and gene sequencing confirmed that the pdl6-95/ANTI-TIMP-1/neo and pdl6-95/siRNA-TIMP-1/neo had been reconstructed successfully. After they had been packed in 293 cells to form rAAV/ANTI-TIMP-1/neo and rAAV/siRNA-TIMP-1/neo, they were used to infect HSC-T6. Thirty days after the infection, the transcription level of TIMP-1 in HSC-T6 cells infected by rAAV/siRNA-TIMP-1/neo decreased dramatically compared with the mock control and normal HSC-T6 cells (P less than 0.01), and the expression level of TIMP-1 gene in HSC-T6 cells decreased significantly (60%), while the transcription and expression level of TIMP-1 in HSC-T6 cells infected by rAAV/ANTI-TIMP-1/neo had no significant difference with mock control and normal HSC-T6 cells (P more than 0.05).</p><p><b>CONCLUSION</b>RNA interference can exert a suppression of TIMP-1 gene in rat HSC, and when this function combines with AAV infection, it can suppress the specific gene expression for a long time by chromosomal integration.</p>
الموضوعات
Animals , Rats , Cells, Cultured , Dependovirus , Genetics , Genetic Vectors , Hepatic Stellate Cells , Metabolism , RNA, Antisense , Genetics , RNA, Small Interfering , Tissue Inhibitor of Metalloproteinase-1 , Metabolismالملخص
<p><b>OBJECTIVE</b>To develop a diagnostic model comprising clinical and serum markers for assessing HBV-related liver fibrosis.</p><p><b>METHODS</b>270 chronic hepatitis B patients were randomly allocated to either an estimation group (195 cases) or a validation group (75 cases). Liver biopsies were done and staging of fibrosis was assessed. Twenty-six common clinical and serum markers were analyzed initially in the estimation group to derive a predictive model to discriminate the stages of fibrosis. The model created was then assessed with ROC analysis. It was also applied to the validation group to test its accuracy.</p><p><b>RESULTS</b>Among 13 variables associated with liver fibrosis selected by univariate analysis, age, gamma glutamyltranspeptidase (GGT), hyaluronic acid (HA), and platelet count (PLT) were identified by multivariate logistic regression analysis as independent factors of fibrosis. A fibrosis index constructed from the above four markers was established. In ROC analysis, the AUC was 0.889 for the estimation group and 0.850 for the validation group for discriminating > or =S3 from < or=S2. Using the optimal cutoff score 3.0, the sensitivity of the index was 90.2%, the specificity 76.1%, and the accuracy was 82%. There was a positive linear relationship between the index scores and the fibrosis stages (r = 0.731, P<0.001). The AUC for identifying > or=S2 was 0.873 with sensitivity/specificity of 79%/82%, cutoff score 2.2; The AUC for identifying S4 was 0.872 with sensitivity/specificity of 83%/75%, cutoff score 5.4. There were no significant differences in diagnostic efficacy in the model between the estimation and the validation group (P>0.05).</p><p><b>CONCLUSION</b>A model for assessment of liver fibrosis was established with easily accessible markers. It appears to be sensitive, accurate and reproducible, suggesting it could be used to assist or replace liver biopsy to detect dynamic changes of HBV-related liver fibrosis.</p>
الموضوعات
Adolescent , Adult , Aged , Female , Humans , Male , Middle Aged , Forecasting , Hepatitis B, Chronic , Diagnosis , Liver Cirrhosis , Diagnosis , Logistic Modelsالملخص
<p><b>OBJECTIVE</b>To investigate the malignant tendency of liver basophilic cells in rats by examining the liver dynamic pathological changes during the hepatocarcinogenesis induced by diethylnitrosamine (DEN).</p><p><b>METHODS</b>One hundred forty male Sprague-Dawley rats, about 200 g each, were randomly divided into a normal group and a model group. The model group rats were administered 1% DEN intragastrically once a week for 14 weeks. The normal control group rats were given saline instead of DEN. Seven to ten rats of the model group were sacrificed at 2, 3, 5, 8, 10, 12, 14, 18 weeks. The remaining rats were followed to the end of the experiment at 26 weeks. Histopathological changes of the livers were analyzed, and the localization of GST-P and PCNA in the livers were detected in situ by immunohistochemistry.</p><p><b>RESULTS</b>According to the characteristics of the lesions in the model group, histological liver change patterns were categorized into three phases: (1) liver injury phase (2 to 5 weeks) with centrilobular necrosis, a small amount of collagen deposition in the necrotic regions with fibrous septa development and cell proliferation; (2) the cirrhosis phase (8 to 12 weeks) with significant hepatocellular regeneration and collagen deposition. As the regenerative nodules and fibrous septa formed, cirrhosis with uniform sized nodules developed in all the rats at 12 weeks. In the regenerative nodules, significant hepatocellular metamorphosis was seen; (3) In the carcinomatous transformation and nodular remodeling phase (after 14 weeks), two types of cancer, namely hepatocellular carcinoma and cholangiocarcinoma were found. Incidence of the cancer was 62.5% at 18 weeks. Basophilic cell lesions appeared beginning at 10 weeks. Pale bodies were seen in some basophilic cells. Small cell changes appeared starting at 12 weeks. Some of these cells containing droplets like lipid vacuoles, invaded into the surrounding liver tissues. Both basophilic cell lesions and small cell changes were all positive for proliferating cell nuclear antigen (PCNA).</p><p><b>CONCLUSION</b>Development of foci of basophilic small cells, with cells containing lipid vacuoles and pale bodies, and invading into the surrounding liver tissues are the changes highly suggestive of an early hepatocellular carcinoma transformation.</p>