Objective To study the expression and its significance of Platelet-derived growth factor(PDGF) and Collagen type Ⅰ (Col- Ⅰ) mRNA in hepatic tissue in mice exposed to arsenic in drinking water. Methods Fifty mice were divided into control group, sodium arsenite group(iAs3+ group, 300 mg/L) and sodium arsenate group (iAs5+ group, 300mg/L) at random. The mice were sacrificed after 10 months for liver function test (ALT, AST and GLB examined by automatic biochemical analyzer) and pathologic examinations. Masson staining and immunohistochemical SABC methods were used. Through the result of Masson staining fibrosis semi-quantitative analysis of the fibrosis number was carried out in the per unit area. Using immuonhistochemical SABC methods, the expression of PDGF and Col- Ⅰ was detected. Results ①Serum ALT was statistically significant among control group [(36.67±3.51) U/L], iAs3+ group[(61.46±13.85) U/L] and iAs5+ group[(43.31±4.21) U/L, F= 6.56, P < 0.05], especially between the control and iAs3+ group (P < 0.05) ;Serum AST had a statistical significance among control group [(135.00 ± 20.42)U/L], iA3+ group [(510.86 ± 59.01)U/L] and iAs5+ group[(258.93 ± 22.40) U/L, F= 83.36, P < 0.05] ;Serum GLB had no statistical significance among control group [(20.86 ± 0.61)g/L], iAs3+ group [(26.94± 3.73)g/L] and iAs5+ group[(24.59 ± 5.27)g/L, F = 2.80, P < 0.05]. ②Masson staining revealed notable liver cell necrosis, regeneration, infiltration of inflammatory cells in portal area. Masson staining Semi-quantitative results showed significantly increasing size of fibrosis in model group. Compared with control group(0.069 ± 0.013) and iAs5+ group(0.143 ± 0.122), fiber hyperplasia of iAs3+ group(0.192 ± 0.108) had statistical significance (F = 1.91, P < 0.05). ③Immunohistochemistry results showed higher expression of PDGF in the portal area, sepetal and preisinus cells in model group. The heavy expression of Col- Ⅰ distributed in the surrounding of blood vessels and bile duet and along portal wall extensting to the liver in model group. Semi-quantitative results showed that PDGF among control group(202.79 ± 7.46), iAs3+ group(174.38 ± 7.71) and iAs5+ group(177.64 ± 7.81) had a statistical significance(F= 102.91, P < 0.05);Col-Ⅰ had a statistical significance among control group(200.11 ± 7.46), iAs3+ group (176.47±10.20) and iAs5+ group (177.378 ±7.948, F = 78.51, P < 0.05). Conclusions Exposed in oral drinking arsenic, mice can develop hepatic injury and hepatic fibrosis. The PDGF and Col-Ⅰ play significant effects on the process of hepatic fibrosis caused by oral drinking arsenic exposed mice.

OBJECTIVE:To study the advantage of microwave technology in the preparation of plastic beads. METHODS:Three kinds of representative processing methods were applied to prepare Equus asinus,Colla Cornus Cervi and Colla Carapax et Plastri Testudinis. The processing methods and the quality of plastic beads were compared. RESULTS:Study results showed mic-rowave technology was superior to other processing methods in the preparation of plastic beads and quality. CONCLUSION:The microwave technology has its specific advantages in the preparation of plastic beads.

Objective To study epidemiology, clinical findings, diagnosis and treatment of sepsis caused by Vibrio vulnificus. Method Patientss with Vibrio vulnificus sepsis were collected from 1995 to 2008. The medical records including epidemiological and clinical data were analyzed. Results The male-to-female ratio of 34cases was 4.7:1 and 76. 5% of these patients suffered from chronic liver disease. Most patients occurred from April to October with signs of abrupt fever, characteristic cutaneous lesions, hypotension and progressive multiple organ disfunction syndrome (MODS). The mortality was over 47.1% . The criteria proposed for early diagnosis of Vibrio vulnificus sepsis were abrupt onset with fever during the period from April to November, characteristic cutaneous lesions, such as the most commonly occurred haemorrhagic bullae on the extremities or even extensive necrosis of skin and muscular tissue, progressive hypotension or shock accompanied by MODS, pre-existing liver disease or chronic abuse of alcohol, and consumption of raw seafood or exposure to seawater within 12 week. Early administration of the third-generation cephalosporins with the quinolones in full dosage, aggressive wound debridement,appropriate dermoplasty and supportive care contribute to a better outcome. Conclusions Vibrio vulnificus sepsis progresses rapidly with high mortality. Early diagnosis, rapid treatment with prompt antibiotics and aggressive surgery treatment are very important to improve the outcome.

Objective To investigate the effect and mechanism of liver X receptor ( LXR ) agonist on expression of fatty acid synthase( FAS) in diabetic kidney. Methods In the part of in vivo study, immunostaining was used to detect the FAS protein expression in kidney. 16-week-old male db/db mice on C57BL/6 background were administered via gavage a LXR synthetic agonist, TO901317, at a dose of 3 mg · kg-1 · d-1 or vehicle ( 0. 5%Carboxymethyl Cellulose Sodium, CMC-Na) alone for 7 d;Quantitative RT-PCR and Western blot were used to detect mRNA and protein levels of FAS and SREBP-1. In the part of in vitro study, MCT cell(a mouse murine proximal tubule cell line)was treated with 10μmol/L TO901317 for 24 h or transfected with active SREBP-1c expression vector (SREBP-1cN). HEK293 cells(a human renal tubule cell line)were transfected with mFAS-(1. 7 kb)-luc, LXR expression vector or SREBP-1cN for 12 h. Quantitative RT-PCR and luciferase reproter assay were utilized to examine FAS mRNA level and FAS promoter activity. Results FAS was abundantly expressed in renal cortex, with low expresson in renal glomeruli. The mRNA and protein expressious of FAS in kidney of db/db mice were lowered compared with db/m mice. TO90137 treatment increased FAS mRNA expression by 1. 3-fold. TO901317 increased expression of SREBP-1 in kidneys of db/m and db/db mice by 5. 1-fold and 17-fold, respectively. TO901317 and overexpression of SREBP-1c increased expression of FAS in MCT cells by 1. 5-fold and 1. 8-fold. Transcription activity of FAS were induced by TO901317, LXR, and SREBP-1cN overexpressions in HEK293 cells. Conclusions Both direct(LXRE)and indirect(SREBP-1c)mechanisms may contribute to the up-regulation of FAS expression by LXR in renal proximal tubule cells.

Objective To evaluate the early clinical efficaoy of hybrid procedure for infants less than six months old with ventricular septal defect and coarctation of aorta.Methods From January 2010 to July 2011,20 patients with ventricular septal defect and coarctation of aorta received hybrid procedure in our center.The body weight was (4.5 ± 1.6) kg ( ranged from 1.9 kg to 6.5 kg) and the age was ( 56 ± 45 ) days ( ranged from 18 days to 6 months).The pressure gradient of the coarctation of the aorta ranged from 30 mm Hg to 56 mm Hg,5 patients of them were diagnosed as hypoplasty of aortic arch.The size of the ventricular septal defect ranged from 8 mm to 16 mm.Results The mortality was zero in all the 20 cases during the surgery,and the mobidity was 20％ (4/20).The complications were pneumonia in 2 cases,infective endocarditis in 1 case and pneumothorax in 1 case.The diameter of coarctation of the aorta ranged from 1.5 mm to 3.4 mm,and the size of the balloon ranged from 4 mm to 12 mm.The pressure gradient of the coarctation of the aorta decreased to 0 to 27 mm Hg.The bypass time ranged from 40 minutes to 87 minutes,and the crossclamp time of the aorta ranged from 20 minutes to 41minutes.The atrial septal defects were repaired and the patent ductuses were ligated during the surgery without leaving the sternum open.The total operation time was (4.0 ± 0.7 ) hours ( ranging from 3.0 hours to 5.2 hours).The mean ventilation time was (2.2 ± 1.4) days and mean ICU stay time was (5 ± 3 ) days.All the patients were followed up for ( 10.0 ± 3.6) months without aneurysm in arch and obstruction in airway.The residual obstructive pressurc gradicnt in the aortic arch ranged from 12 mm Hg to 35 mm Hg and 2 patients received reintervention.One patient received re-balloon dilation and the other received surgery.The cardiac function reached NYHA Ⅰ - Ⅱ in all eases.Conclusion The early outcome of the hybrid procedure (balloon dilation of the coartation of the aorta and surgical repair of ventricular septal defect) for infants with ventricular septal defect and coarctation of aorta was satisfying,which could avoid from circulatory arrest.It is a relatively safe procedure which could be the optional method for one-stage surgical repair.

A total of 41 patients with aspiration pneumonia after nasopharyngeal carcinoma radiotherapy were retrospectively selected from January 2005 to December 2010.They were divided into early bronchoscopy group (n =24) and conventional therapy group (n =17) to analyze the therapeutic effects of early bronchoscopy on temperature,white blood cell (WBC),absorption of chest radiography and mortality rates.The temperature and WBC were at the same level between both groups at pre-treatment [(38.7 ±0.7)℃ vs.(38.5 ±0.7)℃,P=0.633; (15.8 ±4.2) × 109/L vs.(16.2 ±3.4) × 109/L,P =0.430]while the temperature declined obviously after a 3-day treatment [(37.3 ±0.9)℃ vs.(38.4 ± 1.4)℃,P =0.015] and also WBC after a 5-day treatment[(10.6±4.2) × 109/L vs.(15.3 ±6.9) × 109/L,P=0.045].The bronchoscopy group had a faster absorption of chest radiography (Z =-3.515,P =0.00).The mortality rate showed no statistically significant difference between both groups.

Objective To identify the differential serum proteins in patients with hepatic injury resulting from coal-burning type of arsenism. Methods Six serum samples were collected from patients with liver injury resulting from coal-burning type of arsenism and healthy subjects(control gruop) in endemic arsenism area. Twodimensional gel electrophoresis(2-DE) was performed to separate serum proteins, after silver staining, the differential expression of proteins were analyzed and then identified by matrix-assisted laser desorption/ionization time of flight mass spectrometry(MALDI-TOF-MS). Results The 2-DE map of serum protein patterns of patients and normal control were established successfully. The results showed that there were an average of (824 ± 31 ) spots and (782 ± 42) spots on 2-DE matching of the patients and control groups and the matching rate was 94.9%(782/824). From these two groups 49 differential protein spots were identified, of which over 3 times the difference in the expression of 30 protein spots were singled out and MALDI-TOF-MS analysis was carried out. Ten proteins were identified. Upregulated expression was observed in alpha-2-macroglobulin, B-cell receptor-associated protein, keratin 1,apolipoprotein A-I, and down-regulated expression was observed in haptoglobin, α2-heremans-schimid-glycoprotein,mitogen-activated protein kinase 4, zinc finger protein 323, ZAP-70 and SP40 in the patient group. Conclusions The well-resolved and reproducible 2-DE serum patterns of patients are established and some differentially expressed proteins are characterized. Whether these proteins of differential expression are serum markers for liver injury resulting from coal-burning type of arsenism need to be further verified.

Objective To isolate and identify exosomes from serum samples of the patients with polymyositis / dermatomyositis (PM/ DM),and analyze their protein composition preliminarily.Methods Exosomes from serum samples of the patients with PM/DM were isolated and purified by the ExoQuickTM kit.The morphological characteristics and particle size of exosomes were determined by transmission electron microscope (TEM) and NanoSight analyzer,respectively.The surface markers of exosomes such as CD9,CD81 and Flotillin-2 were identified by western blot.The concentration and composition of exosome protein were determined by the BCA method and SDS-PAGE,respectively.Results The exosomes from serum samples of PM/DM patients displayed round or oval vesicles with membrane structure under TEM,and their diameter range was about (92 ± 67) nm.western blot showed that these exosomes expressed CD9,CD81 and Flotillin-2.The total protein concentrations of exosomes in the patients with PM/DM and healthy controls were 14.68 (6.00,32.55) μg/μL and 14.09 (8.00,23.28) μg/μL,respectively.SDS-PAGE showed that high-abundance proteins enriched in 55-70 kD in both PM/DM patients and healthy controls,and that there were different bands in 40-55 kD between them.Conclusion Exosomes are isolated from serum samples of the patients with PM/DM successfully,and their protein concentration and composition are analyzed preliminarily,which provides the experimental evidences for further finding differential proteins.

[ ABSTRACT] AIM:To study the effect of livin gene-modified bone marrow mesenchymal stem cells ( BM-MSCs) transplantation on the cardiac function following acute myocardial infarction in a rat model and the expression of livin , caspase-3, caspase-7 and caspase-9 in the livin gene-modified BM-MSCs.METHODS: The MSCs were obtained by the whole bone marrow culture method , and the apoptosis of the MSCs after infection with adenovirus vector carrying enhanced green fluorescent protein ( EGFP) gene and livin recombinant vector ( rAd-livin) were detected by flow cytometry .The ex-pression of livin, caspase-3, caspase-7 and caspase-9 was detected by Western blot .After permanent left anterior descend-ing artery occlusion , the rats were randomized to receive intramyocardial injection of DMEM without cells ( vehicle group ) , or containing MSCs ( MSCs group ) , MSCs ( EGFP ) ( rAd-control/MSCs group ) or MSCs ( livin ) ( rAd-livin/MSCs group).Left ventricular systolic pressure (LVSP), left ventricular end-diastolic pressure (LVEDP), the maximum in-creased rate of left ventricular pressure ( -dp/dtmax ) and the maximum decline rate of left ventricular pressure ( +dp/dtmax ) were recorded for evaluating the cardiac functions .RESULTS: The apoptosis of rAd-livin/MSCs was significantly decreased as compared with MSCs and rAd-control/MSCs (P<0.05).Meanwhile, the expression of caspase-3, caspase-7 and caspase-9 was significantly downregulated as compared with the other 2 groups ( P<0.05 ) .The cardiac function in rAd-livin/MSCs group was significantly improved as compared with DMEM group , and those in the other 2 groups got the similar results, but the function in rAd-livin/MSCs group was better improved .Meanwhile, the number of surviving cells in rAd-livin/MSCs group was significantly improved as compared with the other 2 groups .CONCLUSION:The apoptosis of MSCs is decreased after rAd-livin transfection, and the expression of caspase-3, caspase-7 and caspase-9 is also significant-ly downregulated while the expression of livin is significantly upregulated .Transplantation of livin-modified BM-MSCs by lentiviral vector results in better prognosis for treating myocardial infarction by enhancing cell survival .

OBJECTIVETo construct a recombinant plasmid pIRES-GM-CSF-IL-21, and to investigate its antitumor effect on tumors in the mice.

METHODSFifty Bal b/c mice were included in this study. Cultured hepatoma H22 cells were inoculated in the left lobe of the liver to develop orthotopically transplanted liver tumor models. The mice with orthotopically transplanted liver tumor were randomly divided into 5 groups (n = 10): (1) Each mouse received injection of recombinant plasmid pIRES-GM-CSF-IL-21; (2) Received injection of plasmid pIRES-GM-CSF; (3) pIRES-IL-21; (4) Received injection of ampty plasmid pIRES (H22/neo group); (5) Received injection of PBS (H22 group) via the tail vein, respectively. Therefore, the anti-tumor effect was induced by both GM-CSF and IL-21, or by either of them alone. The serum levels of IFN-γ and IL-2 were detected by ELISA, and the cytotoxicity of spleen NK and CTL cells were tested by MTT colorimetry.

RESULTSComparing with the H22 and H22/Neo groups, the tumor weight in the mice of H22/GM-CSF group was (0.603 ± 0.223) g, H22/IL21-treated group (0.583 ± 0.290) g and H22/GM-CSF-IL21-treated group (0.303 ± 0.323) g, significantly lower than that in the H22 group [(1.591 ± 0.280) g] and H22/Neo group [(1.489 ± 0.155) g]. Among them the tumor growth was most significantly inhibited in the H22/GM-CSF-IL-21 group (0.303 ± 0.323) g, compared with that of H22 and H22/neo groups (P < 0.01). But there was no significant difference between the tumor weights of the H22/GM-CSF group and H22/IL-21 group, and between the tumor weights of the H22 and H22/Neo groups (P > 0.05). The levels of IFN-γ and IL-2 in peripheral blood of mouse models treated with H22/GM-CSF-IL-21 were significantly increased than that in the H22/GM-CSF group and H22/IL-21 group (all P < 0.01), but significantly decreased in the H22group and H22/Neo group (P < 0.01). The anti-tumor activity of splenic NK cells and CTLs in the H22/GM-CSF-IL21 group was significantly enhanced (P < 0.01), compared with the significantly decreased in the H22 and H22/Neo groups.

CONCLUSIONSOur results demonstrate apparent antitumor effect of the plasmid pIRES-GM-CSF-IL-21 on the orthotopically transplanted liver tumor in mice. The combination of both pIRES-GM-CSF and IL-21 is more effective than that of pIRES/IL21 or pIRES/GM-CSF treatment alone. In addition, the plasmid pIRES-GM-CSF-IL-21 can also promote the secretion of IFN-γ and IL-2 in vivo, and enhance the cytotoxic activity of splenic NK and CTLs against the transplanted liver tumor.

Animals ; Carcinoma, Hepatocellular ; blood ; pathology ; therapy ; Cell Line, Tumor ; Female ; Granulocyte-Macrophage Colony-Stimulating Factor ; genetics ; Immunotherapy ; Interferon-gamma ; blood ; Interleukin-2 ; blood ; Interleukins ; genetics ; Killer Cells, Natural ; immunology ; Liver Neoplasms ; blood ; pathology ; therapy ; Mice ; Mice, Inbred BALB C ; Neoplasm Transplantation ; Plasmids ; therapeutic use ; Random Allocation ; Recombinant Proteins ; therapeutic use ; T-Lymphocytes, Cytotoxic ; immunology ; Tumor Burden