In order to enhance the specificity of TNF-α monoclonal antibody to inflamed site, a bispecific antibody BsDb that targets TNF-α and the extra-domain B (ED-B) of fibronectin (FN) was constructed by covalently linking the anti-TNF-α single chain Fv antibody (TNF-scFv) and the anti-ED-B scFv L19 via a flexible peptide linker deriving from human serum albumin (HSA). ED-B is an antigen specifically expressed at the inflamed site. BsDb is expressed in E. coli, identified by immunoblot, and purified with affinity chromatography. This was followed by further examination of its bioactivities and pharmacokinetics. We demonstrated that BsDb retained the immunoreactivity of its original antibodies as it could simultaneously bind to TNF-α and ED-B and neutralize the biological action of TNF-α. In the collagen-induced arthritis mice model, BsDb selectively accumulate in the inflamed joint with a maximal uptake of (12.2 ± 1.50)% ID/g in a single inflamed paw and retain in the inflamed paw for at least 72 h. In contrast, BsDb showed a short serum half-life of (0.50 ± 0.05) h and a rapid clearance from normal tissues. The findings reported herein indicate that BsDb has good specificity to the inflamed site and low toxicity to normal tissues. BsDb is therefore likely to have greater clinical applications in the treatment of rheumatoid arthritis and other autoimmune diseases. This laid a stable basis for its preclinical study.
Animals ; Antibodies, Bispecific ; chemistry ; Antibodies, Monoclonal ; chemistry ; Arthritis, Experimental ; Escherichia coli ; Fibronectins ; chemistry ; Half-Life ; Humans ; Mice ; Single-Chain Antibodies ; chemistry ; Tumor Necrosis Factor-alpha ; chemistry

BACKGROUNDPeritoneal dissemination is the most common pattern of metastasis in advanced gastric carcinoma with serosal invasion. In the present study, we reported the clinical relevance of a new diagnostic method involving RT-PCR, using survivin as the target gene, for the detection of free cancer cells in peritoneal washes.

METHODSIntraoperative peritoneal washes were obtained from 48 patients who underwent surgery for gastric cancer. RT-PCR analysis with primers specific for survivin and conventional cytological examinations were both performed.

RESULTSSurvivin mRNA was not detected in any peritoneal wash samples from patients with benign disease, but was detected in 28 of 48 samples taken from patients with gastric cancer and in all metastatic nodules. Survivin expression in the peritoneal cavity significantly correlated with depth of cancer invasion, lymph node metastasis, and TNM stage. There were 92% of clinically evident peritoneal metastasis cases showed detectable survivin expression. The combination of survivin RT-PCR and cytological examination yielded positive results in 66.7% (32/48) of patients with gastric cancer, much higher than the results produced by cytological method alone.

CONCLUSIONSSurvivin mRNA detected in peritoneal lavage fluid might indicate the presence of free cancer cells in the peritoneal cavity. The high sensitivity of the RT-PCR-based survivin assay suggests that survivin serves as a molecular marker for detecting peritoneal micrometastasis. Its ubiquitous expression in peritoneal cancer cells and metastatic nodules also suggests a promising future therapeutic strategy based on survivin inhibition for cases of gastric cancer involving peritoneal metastasis.

Adult ; Aged ; Aged, 80 and over ; Ascitic Fluid ; metabolism ; Female ; Humans ; Inhibitor of Apoptosis Proteins ; Male ; Microtubule-Associated Proteins ; genetics ; Middle Aged ; Neoplasm Proteins ; Neoplasm Staging ; Peritoneal Neoplasms ; secondary ; RNA, Messenger ; analysis ; Reverse Transcriptase Polymerase Chain Reaction ; Stomach Neoplasms ; metabolism ; pathology

Objective To study the prevalence and grade of cerebral microbleeds (CMB) among patients with different subtypes of acute ischemic cerebrovascular diseases, and investigate the clinical significance of CMB.Methods Consecutive 259 patients with acute ischemic cerebrovascular diseases, admitted to our hospitals from September 2009 to July 2010, were included; according to the stroke subtypes, these patients were classified into groups of atherothrombotic infarction (n=146),cardioembolic infarction (n=28), small artery infarction (n=50), infarction of unknown origin (n=19) and transient ischemia attack (TIA, n=16). The patients without cerebral vascular diseases were served as controls (n=96). The baseline data were registered and all patients were performed gradient echo-T2*weighted imaging (GRE-T2*WI); the prevalence and grade of CMB between each 2 different subtypes of acute ischemic cerebrovascular diseases were compared; the prevalence of CMB in patients with acute ischemic infarction for the first time and patients with recurrent cerebral infarction was compared.Results The prevalence and grade of CMB between each 2 different infarction subtypes varied with a statistical difference (P＜0.05). Apart from that of TIA group, the prevalence of all infarction groups was statistically higher than that of the controls (P＜0.05) with small artery infarction group being the highest (68.0%). The prevalence of CMB in patients with recurrent infarction was statistically higher than that in patients with primary infarction (P＜0.05).Conclusion The prevalence of CMB among different subtypes of infarction is high with the subtype of small artery infarction enjoying the highest rate; the prevalence of CMB in recurrent infarction goes higher as compared with that in primary infarction; the relapse of the cerebral infarction is possiblely related to the presence of CBMs.

Female ; Humans ; Lymphatic Metastasis ; Male ; Microdissection ; Neoplasm Invasiveness ; RNA, Messenger ; analysis ; Reverse Transcriptase Polymerase Chain Reaction ; Stomach Neoplasms ; genetics ; pathology ; rho GTP-Binding Proteins ; genetics ; rhoC GTP-Binding Protein

OBJECTIVEProbe the effects of rhGH on severe degree burned patients' blood sugar in different age of years.

METHODSElected 210 patients hospitalized in the Third Affiliated Hospital of Wenzhou Medical College from January 2005 to December 2008, who were burned in 48 h, older than 18 years, ever had no diabetes and tumor history and placidly pull through shock stage. Among the patients there were 132 males and 78 females. The age was from 18 to 65 years old, average (40.7 +/- 7.2) years old. The extent of burn were form TBSA 25% to TBSA 86%, average TBSA (40.4 +/- 12.5)%. The depths of burn were from superficial second degree to third degree. All of the total divided into A (18 - 44 years old) and B (> 45 years old)groups. Each group had 105 patients. Two groups were randomly divided into A(1), A(2), A(0) and B(1), B(2), B(0) groups. Each group had 35 patients. The A(1) and B(1) groups were used 0.15 U/(kg.d) growth hormone (Somatropin, S19990021), A(2) and B(2) groups were used 0.2 U/(kg.d) growth hormone, A(0) and B(0) groups were used NS as control. Observed and analyzed the change of blood sugar and insulin amount used in 210 patients.

RESULTSOf all the patients in 6 groups, there were 190 patients finished the experimentation in four weeks. The insulin amount of A(1), A(2), A(0) groups used were (2123.3 +/- 152.3), (2885.6 +/- 148.5), (724.1 +/- 31.1) U, B(1), B(2), B(0) group were (2715.1 +/- 95.3), (3652.2 +/- 198.1), (801.8 +/- 22.2) U. The consequence showed that the number need insulin to control blood sugar in B group was more than A group, as well as using 0.2 U/(kg.d) does to 0.15 U/(kg.d) does, and using growth hormone to no using(P < 0.01). The time that blood sugar of A(1), A(2), B(1), B(2) group recovered to normal range without using insulin were (5.11 +/- 0.82), (4.93 +/- 0.89), (5.2 +/- 0.65), (5.13 +/- 1.02) d (P > 0.05).

CONCLUSIONSThe blood sugar's alteration has positive correlation with the age of years and the does of rhGH. As long as normative using rhGH it doesn't induce diabetes.

Adolescent ; Adult ; Age Factors ; Aged ; Blood Glucose ; drug effects ; metabolism ; Burns ; blood ; drug therapy ; Dose-Response Relationship, Drug ; Female ; Human Growth Hormone ; administration & dosage ; therapeutic use ; Humans ; Male ; Middle Aged ; Young Adult

OBJECTIVETo propose a new method for dynamic positron emission tomographic (PET) image reconstruction using low rank and sparse penalty (L&S).

METHODSThe L&S reconstruction model was established and the split Bregman method was used to solve the optimal cost function. The one-tissue compartment model was used to simulate a set of PET 82Rb myocardial perfusion image. The L&S reconstruction method was compared with maximum likelihood expectation maximization (MLEM) method, low-rank penalty method and sparse penalty method.

RESULTSThe L&S reconstruction method had the smallest MSE and well maintained the feature information. The polar map created by L&S method was the most similar with the reference actual polar map.

CONCLUSIONL&S reconstruction method is better than the other three methods in both visual and quantitative analysis of the PET images.

Algorithms ; Humans ; Likelihood Functions ; Positron-Emission Tomography ; methods

Objective:To investigate the renal protective effect of Tangshenping capsule (Tangshenping) on diabetic nephropathy (DN) KKAy mice and its effect on Wnt/β-catenin signaling pathway. Method:Sixty female Sprague-Dawley KKAy mice aged 10 weeks old were induced with KKAy rat feed for 10 weeks. The DN animal model was successfully determined with blood glucose (>16.7 mmol·L^-1) and 24 hour urine protein (>0.4 mg). The model mice were randomly divided into a model group, an irbesartan group, and low, medium and high-dose Tangshenping group, with 10 female C57BL/6J mice as a control group. The treatment groups were given the corresponding drugs by gavage. The normal group and the model group were given an equal volume of deionized water by gavage. The intragastric dose was 0.01 mL·g^-1 body weight coefficient once a day. The general conditions of the mice were observed, the body mass was weighed every 4 weeks, and 24 h urine protein was quantified. At the 26^th week, the blood was collected from eyeballs, and the mice were put to death. The quality of the kidneys, serum blood urea nitrogen (BUN), serum creatinine (SCr), triglyceride (TG), malondialdehyde (MDA), nitric oxide (NO) and superoxide dismutase (SOD) content were measured. In situ hybridization and immunohistochemistry were used to detect the expressions of Wnt4, glycogen synthase kinase 3β(GSK3β) and β-catenin in kidney tissues. Result:Compared with model group, body mass, kidney mass/body mass, and 24 h urine protein were significantly lower in high-dose Tangshenping group (P<0.01). The pathological damage of the kidney was significantly reduced, the serum BUN, SCr, TG and MDA contents were significantly decreased (P<0.01), the contents of NO and SOD were significantly increased (P<0.01), and the protein and mRNA expression levels of Wnt4, Gsk-3β and β-catenin were decreased (P<0.01). Conclusion:Tangshenping may inhibit the activation of Wnt/β-catenin signaling pathway, reverse the transdifferentiation of renal tubular epithelial cells in DN KKAy mice, delay the progression of renal interstitial fibrosis, and then exert renal protection.

OBJECTIVE: To develop an automated chimeric analysis and reporting platform based on short tandem repeat (STR) and capillary electrophoresis methods for allogeneic hematopoietic stem cell transplantation (allo-HSCT) so as to improve work efficiency. METHODS: Apache, MySQL, PHP and HTML5 were used to build the database and interface. The STR locus geno typing and chimeric analysis logic and flow were set up on the basis of STR rules and capillary electrophoresis. STR genotyping and 194 times of chimeric testing data of 100 patients after allo-HSCT were used to test the platform for automatic STR locus genotyping, chimeric calculation and report generation. RESULTS: The established platform could realize the functions of STR locus customization, STR genotype determination, automatic chimeric analysis, and detection information database management, which can automatically generate an integrated report including multiple sequential chimeric results and trend graphs for the same patient and can be accessed and used simultaneously by different users through different browser interfaces. The results of automated analysis by the platform are completely consistent with that of manual analysis by experienced technicians, and the possibility of manual analysis error is reduced through automation. The time required for automatic analysis using this platform is approximately 1/6-1/5 of manual analysis. CONCLUSION The automatic analysis platform built in this study is operation stable and reliable in analysis results, which can improve work efficiency and report connotation, thus worthing popularized and applicable.
Electrophoresis, Capillary ; Genotype ; Hematopoietic Stem Cell Transplantation ; Humans ; Microsatellite Repeats ; Tissue Donors

Adult ; Aged ; Alanine Transaminase ; blood ; Cardiovascular Diseases ; blood ; epidemiology ; China ; epidemiology ; Cross-Sectional Studies ; Female ; Humans ; Male ; Metabolic Syndrome ; blood ; epidemiology ; Middle Aged ; Risk Factors

OBJECTIVE: The association between lipoprotein (a) [Lp(a)] levels and metabolic syndrome (MetS) remains uncertain, especially in the Asian population. The purpose of this study was to demonstrate the association between Lp(a) levels and MetS in a middle-aged and elderly Chinese cohort. METHODS: A cross-sectional study of 10,336 Chinese adults aged 40 years or older was conducted in Jiading District, Shanghai, China. Logistic regression analysis was used to evaluate the association between serum Lp(a) levels and MetS. RESULTS: In the overall population, 37.5% of participants had MetS. Compared with individuals in the lowest quartile of serum Lp(a) levels, those in the highest quartile had a lower prevalence of MetS (30.9% vs. 46.9%, P for trend < 0.0001). Multivariate logistic regression analyses showed that compared with participants in the bottom quartile of serum Lp(a) levels, those in the top quartile had decreased odds ratio (OR) for prevalent MetS [multivariate-adjusted OR 0.45 (95% confidence interval 0.39-0.51); P < 0.0001]. Additionally, Lp(a) level was conversely associated with the risk of central obesity, high fasting glucose, high triglycerides, and low HDL cholesterol, but not with hypertension. Stratified analyses suggested that increasing levels of Lp(a) was associated with decreased risk of MetS in all the subgroups. CONCLUSION Serum Lp(a) level was inversely associated with the risk of prevalent MetS in a middle-aged and elderly Chinese cohort.
Aged ; Asian Continental Ancestry Group ; China ; epidemiology ; Cross-Sectional Studies ; Female ; Humans ; Lipoprotein(a) ; blood ; Male ; Metabolic Syndrome ; blood ; epidemiology ; Middle Aged