Objective The differences of von Willebrand factor(vWF),hypersensitive C-reactive protein(hs-CRP) and soluble P-selectin CD62P were observed in coronary heart disease(CHD) patients with different syndrome patterns,and the correlation of traditional Chinese medical(TCM) syndrome patterns with endothelial damage,inflammation and platelet activation in CHD patients was investigated.Methods One hundred and eighty-six CHD patients were differentiated as the syndromes of blockage of heart-blood-stasis,phlegm blocking heart vessels,deficiency of heart and kidney yin,deficiency of both yin and Qi,and deficiency of yang and Qi.Thirty healthy volunteers served as normal control.The blood levels of vWF,hs-CRP and CD62P were detected in the CHD patients and the normal controls.Results Blood vWF level in CHD patients was higher than that in the normal controls(P0.05).Blood CD62P in the syndrome pattern of deficiency of yang and Qi was higher than the other patterns(P

Objective To determine the level of adiponectin (APN) in serum and induced sputum of patients with chronic obstructive pulmonary disease both during acute exacerbation (AECOPD) and silent stage, and investigate APN' s role as a marker of inflammation in the pathogenesis of COPD. Method From October 2008 to October 2009,30 male AECOPD patients in the emergency department, 30 male silent COPD patients in the department of respiratory diseases and 30 healthy nonsmoking male volunteers were included. All subjects' serum and induced sputum were collected, and they were all of normal weight(BMI range of 18.5～ 24.9 kg/m2). Patients were excluded if they suffered from severe bronchial asthma, bronchiectasis or autoimmune disease. The number of cells in induced sputum was counted and the cell type was classified. The concentrations of APN, IL-8, IL-6 and TNF-α in both serum and sputum were measured by using ELISA, and their pulmonary function was tested. The different groups were compared among them by using the t -tests, ANOVA analysis or nonparametric analysis, the relation between variables was assessed by using the Pearson or Spearman correlation test. Results The concentrations of APN in both serum and induced sputum of AECOPD patients were significantly higher than those in the silent COPD patients and the control subjects ( P ＜ 0.01 ). The concentrations of APN in the silent COPD patients were significantly higher than those in the control subjects ( P ＜ 0. 01 ). There were significant relationships between the concentrations of APN in serum and induced sputum and the levels of IL-8 and TNF-α in AECOPD patients ( r = 0.739, 0. 734,0.852 and 0. 857, respectively, P ＜ 0. 05) and in silent COPD patients ( r = 0.751,0.659, 0.707 and 0.867, respectively, P ＜0.05). There was significant relation betweenship between APN and neutrophil in induced sputum of AECOPD patients (r = 0.439, P ＜ 0.05). Conclusions APN was involved in the process of systemic and airway inflammation of COPD, and it was related with IL-8 and TNF-α. APN can be used as a new inflammation marker for COPD.

Objective To obtain the short peptides from phage-displayed random peptide library through screening the epitope of monoclonal antibody against tumor necrosis factor(TNF-?). Methods Anti-TNF-? was used to immunoscreen a phage random peptide library of 12 amino-acidresidues displayed as a fusion to protein Ⅲ of filamentous phage M13. The positive clones were obtained by three rounds of biopanning, and the reactivity of each clone binding to anti-TNF-? was examined by double-antibody sandwich ELISA and Dot-ELISA. Mixed positive phage clones were used to detect the serum from SLE patients and healthy persons by Dot-ELISA. Results The eluted phages were enriched nearly 100 fold through three rounds of biopanning, 7 phage clones from the third round biopanning were randomly selected and 5 clones of them could bind to the anti-TNF-?. The binding rate of mixed clones with SLE patients was significantly higher than that of healthy persons. Conclusion The phage display technique can be applied to study the anti-TNF-? antigenic peptides, and these epitopes provide the potential for developing immunodiagnostic reagents of vaccines.

Objective To investigate a rational treatment for severe gallstone pancreatitis(SGP) and evaluate therapeutic effect.Methods A retrospective study was made on the clinical data of 97 cases with SGP.(Results) Among 97 SGP patients,54 cases were in severe grade I, 43 cases were Grade II;77(79.4%) cases were cured and 20(20.6%)died.The morbidity rates of endoscopic therapy(EST),delayed surgery and early surgery were 24.1%,25.0% and 65.5% respecticely,while the mortality rates in the 3 groups were 10.3%,13.9% and 37.5% respectively.The morbidity and morality rates in the first 2 groups were significanty lower than the early surgery group(P

Objective To discuss the value of Visfatin in severity evaluation in patients with severe pneumonia via observation on the variations of the plasma level of Visfatin. Method Seventy subjects including 40 patients with severe pneumonia ( group A) and 30 patients with non-severe pneumonia (group B) admitted to the ICU of emergency department and general wards from June 2009 to June 2010, were enrolled in this prospective study, and another 30 healthy individuals from physical examinees were included as subjects in control group (group C). Patients with severe diseases of heart, brain and kidney, cancers, autoimmune disease, or under special treatment in latest one month were excluded. For the subjects of all three groups, the plasma levels of Visfatin, IL-6, IL-8 and TNF-α were measured by using ELISA, while the level of CRP was assayed by using immunoturbidimetry, and the routine blood test was performed as well. The blood gas analysis and Acute Physiology and Chronic Health Evaluation Ⅱ ( APACHE Ⅱ) were carried out in patients with pneumonia. Comparisons between groups were made by t-tests, ANOVA or nonparametric test. Correlation analysis was carried out by Pearson correlation coefficient or Spearman rank correlation test. Results The plasma level of Visfatin in patients with severe pneumonia (group A) was significantly higher than that in patients with non-severe pneumonia (group B) and in the control subjects (group C) (P < 0. 01) , and the level of Visfatin in pneumonia ( group B) and in control group (group C) , and that in group B was significantly higher than that in the controls (group C) (P <0. 01). In group A, the plasma level of Visfatin was positively correlated with CRP, TNF-α, APACHE Ⅱ and PMN% (rha =0. 653, r = 0.554, r = 0.558, r= 0.484, P <0. 05), while negatively correlated with PaO2 and PaO2/FiO2 ( rha = -0.422, r= -0.543, P <0. 05). Conclusions Visfatin may be involved in the systemic inflammation response in severe pneumonia as a pro-inflammatory cytokine which is valuable in assessing the severity of pneumonia.

Objective To investigate prophylaxis and therapy of early complications following relatives' partial live small bowel transplantation.Methods Four relatives' partial live small bowel transplantations were carried out.Among the 4 patients,there were 3 cases of short intestine syn- drome and one case of non-function of small bowel caused by the absence of nerve ganglion of small in- testine.More than 4 antigens of HLA were completely matched between donators and receptors.In- testines of donators were got from terminal ileum with the length of (150?10) cm.After operations, tacrolimus (FK506),mycophenolate mofetil (MMF),and methylprednisolone were used to prevent rejections.Measures such as use of anticoagulation,improving microcirculation and albumin infusion, aimed at regulating the function of blood coagulation and preventing bleeding and formation of thrombus at anastomotic stoma;famotidine and omeprazole were used to prevent irritable ulcer;use of the third generation of cephalosporins antibiotics,ganciclovir and fluconazol could prevent bacteria,vi- rus and eumycete infections;disinfection and care of easily-infected organs were emphasized;receptors were encouraged to get out of their beds to move frequently;glutamine and enteral nutrition were used early to promote recovery of intestinal function.Results Three days after operation,one patient's lung was infected with baumanii,and the infection had been under control after being treated with the third generation cephalosporins antibiotics;five days after operation,haematoma was detected on an- other patient and was cleared through the second operations growth of eumycete was found in 2 pa- tients' excretion and secretion from enteron,and their situations were improved with fluconazol;acute rejections of the 4 patients were detected 20 days after operation and reversed by the increased use of FK506 combined with methylprednisolone.Among the 4 patients,2 of them have survived for a long time,and the first patient has survived for 6 years and 8 months till now and the other one for 3 years and 2 months;furthermore,other 2 patients respectively died of infections 5 months and 35 days after the operations.Conclusion Because of special constitution of intestine,early complications of rela- tives' partial live small intestine transplantation are frequent and complicated.Therefore,prophylaxis and therapy of early complications are crucial to the success of the transplantation.

Primary liver adenosquamous carcinoma is a rare kind of liver malignant tumor with very poor prognosis. Clinical data of 5 cases were analyzed retrospectively, four male and one female, whose average age was 60.2 years (50-67 years) old. Adenosquamous carcinoma contains both adenocarcinoma and squamous carcinoma histologically, and squamous carcinoma cells are dominant (>80%). Four cases underwent surgical resection and one received liver transplantation. The postoperative follow-up period ranged from 6 to 35 months and the mean survival time of these cases was about 15 months. Three patients died during the follow-up period. The survival rates 1 and 2 years after operation were 60% and 20%, respectively.

[ ABSTRACT] AIM:To cultivate stem-like spheres from SW620 cell line in the specific serum-free medium and evaluate the features of the cancer stem cells, and to investigate the effects of docosahexaenoic acid ( DHA) and eicosapen-taenoic acid ( EPA) on the growth of SW620 stem cell-like cells.METHODS: Human colon cancer stem cell-like cells ( CSCLC) were obtained from SW620 spheres cultured in serum-free medium.These cells were tested for the expression of SSEA-1 and TRA-1-81 by immunofluorescence staining.The mRNA expression of Sox-2 and Oct-4 was detected by real-time PCR.The efficiency of colony formation on a soft agar gel and tumor formation in the nude mice was compared between SW620 adherent cells and CSCLC.The inhibitory effects of 5-fluorouracil (5-FU) and mitomycin C on both types of cells were measured by MTS assay.MTS assay, Annexin V/PI staining and trypan blue staining were used to determine the effects of DHA and EPA on both types of cells.MTS assay was also used to analyze the combined effect of DHA or EPA with chemotherapeutic drugs on SW620 CSCLC.RESULTS:SW620 cells formed spheres in serum-free culture.The cells from spheres highly expressed SSEA-1 and TRA-1-81, transiently expressed Sox-2 and Oct-4 genes and were more resistant to 5-FU and mitomycin C treatments.These cells exhibited a greater ability in clone formation and tumorigenicity, indica-ting that these cells carried stem cell-like features, hence were considered SW620-derived CSCLC.DHA and/or EPA sup-pressed SW620 CSCLC by inhibiting cell growth, inducing cell apoptosis and sensitizing them to chemotherapeutic drugs. CONCLUSION:The cells with stem cell-like features, such as high efficiency in clonogenicity, tumorigenicity and resist-ance to chemotherapeutic drugs, can be obtained from SW620 spheres cultured in serum-free condition.DHA and EPA in-duce apoptosis in SW620-derived CSCLC and sensitize them to chemotherapeutic drugs.

Objective To explore in vitro mesenchymal stem cell (MSC) transfection of lentiviral vector mediated calcitonin gene‐related peptide(CGRP) gene and its effects on biological properties of MSC .Methods MSC were isolated ,cultured and identi‐fied .MSC were infected by lentivirus encoding recombinant enhanced green fluorescent protein (EGFP) gene and CGRP (Lv‐EG‐FP‐CGRP) .The transfection efficiency was determined by the inverted fluorescence microscope and flow cytometry .The expression levels of CGRP were detected in CGRP‐modified MSC by using real‐time PCR ,immunocytochemistry and enzyme‐linked immu‐nosorbent assay (ELISA) .The proliferation ,aging and differentiation ability of MSC were evaluated by MTT ,β‐galactosidase stai‐ning and inducing differentiation respectively .Results After 48 h of MSC transfection by Lv‐EGFP‐CGRP ,EGFP/CGRP could be expressed stably .When multiplicity of infection (MOI) was 30 ,the transfection efficiency reached more than 80% .Compared with the MSC group and the MSC‐EGFP group ,the mRNA and protein expression levels of CGRP in CGRP‐modified MSC(MSC‐CGRP group) were markedly increased(all P<0 .01) .The results of MTT ,β‐galactosidase staining and inducing differentiation assay dem‐onstrated that the transfected CGRP basically had no effect on the proliferation ,aging and endotheliocyte differerntiation of MSC . Conclusion MSC is a kind of ideal genetic vector cell ,which can serve as the target cell of CGRP gene transduction for the applica‐tion of gene therapy and lays the foundation for follow‐up in vitro and vivo experiments .

Objective To evaluate the usefulness of serum galactemannan(GM) for the diagnosis of invasive pulmonary aspergillosis (IPA) in critically ill patients. Methods Study was conducted between February 2007 and July 2008. Included patients on admission ICU who suffer from suspected IPA. GM test and culture were collected 2 weekly. Chnical feature, mycological evidence and optical density index (ODI) were noted. Clinically invasive fungal infection(IFI) were defined proven, probable and possible. The patients were classified into neutropenia, non-neutropenia and treated with immunosuppressive agents, non-neutropenia and non-immunosuppressive agents. To compared of the sensitivity and specificity of GM in different patients. Results 94 patients were included, 4 patients were proven, 29 patients were probable, 34 patients were possible IFI, 27 patients were non-IPA. The positive rate of the GM was 31.9% (30/94). The sensitivity and specificity of GM in proven cases and probable cases are 66.7% and 92.6%. GM assay tended to become positive earlier than the culture 2-10(5.33±2.17)d. We found that differences in patient diagnosis and selection might account for the disparities seen for positive rate for the GM test. There was positive in three of the four patients with proven, the positive rate of GM was 65.5% for probable cases, for possible cases was 17.6%, for non-IPA cases was 7.4% (P=0.001). For patient with neutropenia , treated with immunosuppressive agents and without immunosuppressive agents, the positive rate of GM was 52.9%vs 41.7% vs 34. 6% (P=0.015) ;the sensitivity was 80.0% vs 70. 0% vs 53.8% (P=0.011), the ODI was 1.365 (0.582-6.736) vs 1. 123 (0. 623-6.868) vs 0.554 (0.522-0.823), P=0. 005, respectively. Conclusion These results show that GM test is useful for early diagnosis IPA in critically ill patients. Differences in patient selection and diagnosis might account for the disparities seen for positive rate and sensitivity for the GM test. It has been higher sensitivity and ODI in the patient treated by immunosuppressive agents.