Humanized monoclonal antibodies(mAbs) are increasingly widely used in targeted therapy for cancer and some other major diseases.Complementarity-determining region(CDR) grafting makes quantities of humanized mAbs available.Herein,we provide an overview on the strategy and progress of CDR grafting.

A new flavonoid glycoside, (-)-2S-8-methyl-5,7,4'-trihydroxyflavanone-7-O-β-D-glucopyranoside (1), along with five known ones, quercetin-3-O-(2"-galloyl)-α-L-arabinoside (2), kaempferol-3-O-α-L-arabinoside (3), guaijaverin (4), trifolin (5) and hyperin (6), was isolated from the leaves of Eucalyptus robusta. Their structures with absolute configurations were elucidated by NMR, HR-ESI-MS, CD spectra data and physicochemical methods. In addition, 2-6 were isolated from E. robusta for the first time.
Eucalyptus ; chemistry ; Flavonoids ; chemistry ; isolation & purification ; Glycosides ; chemistry ; isolation & purification ; Plant Leaves ; chemistry

Objective To investigate the role of intestinal fatty acid binding protein(I-FABP)in evaluating intestinal dysfunction of septic rats and the effect of glutamine on I-FABP expression.Methods Rats were divided into 3 groups,control group were only fed with Peptisorb,model group were fed with Peptisorb after intraperitoneal injection with E.coli endotoxin lipopolysaccharidegiven and glutamine group were added glutamine on basis of model group.The correlation between serum I-FABP level and intestinal mucosa damage index were analyzed and the concentrations of serum I-FABP in each group were observed and compared. Results The serum level of I-FABP in rats were correlated with the Chiu’s score of intestinal mucosa,mucosa thickness and villus length(P<0.05 ).Compared with control group,the concentration of serum I-FABP in model group and glutamine group were significantly increased(P<0.05),but which in glutamine group was lower than that in model group(P<0.05).Conclusions Serum I-FABP could be an non-invasive diagnosis index for evaluating acute intestinal dysfunction in septic rats.In addition,dietary glutamine supplementation may ameliorate sepsis-induced intestinal epithelial injury in rats.

Objective To investigate the correlation between sTREM-1 and inflammatory factors expression in critical patients and its effect on severity of disease and clinical prognosis .Methods sTREM-1 ,TNF-α,IL-6 ,IL-10 levels were checked in serum of 54 patients who admitted to intensive care unit (ICU) on the first day and only sTREM-1 was checked again on the third day .At the same time ,APACHEⅡand survival situation in 28 days were recorded .Results sTREM-1 level in critical patients was positive correlated with TNF-α,IL-6 and APACHEⅡ(P<0 .01) .There was positive correlation between APACHE Ⅱ and TNF-α,IL-6 ,but pearson correlation coefficient between sTREM-1 and APACHE Ⅱ was higher than TNF-α,IL-6 .Compared with the survivor group ,the concentrations of serum sTREM-1 was significantly higer in non-survivor group on the first day and the third day after entering ICU(P<0 .05) .Conclusion sTREM-1 level is positive correlated with inflammatory reaction and the severity of disease .It also has prognostic value for outcome in patients with critical illness .

Objective To explore the role of synaptic plasticity on the formation of visceral hyper- sensitivity induced by acute restraint stress in rats.Methods Twenty male Sprague-Dawley rats were randomly divided into control group and acute restraint stress group(model group).Visceral hypersensi- tivity was made by acute restraint stress for 1 h.The colorectal distension(CRD) with different pressure were performed and the abdominal electromyography(EMG) was recorded.The visceral sensitivity was determined by the frequency of EMG.The ultrastrueture of synapse was observed with transmission electron microscope.The expression of synaptophysin was measured by RT-PCR and Western-blot. Results①The frequency of EMG was significantly correlated with CRD pressure(control group,r=0.992, P=0.008;model group,r=0.978,P=0.022).The frequencies of EMG in model group(at 40,60 and 80 mm Hg) were significantly more than that in control group(P value=0.043,0.024,0.038,respectively).②There were more synaptic vesicles accumulated in presynaptical terminal.The post synaptic density was increased in model group compared to control group.③In the proximal and distant colon,the expressions of rnRNA and protein of synaptophysin were higher in model group (P

The aim of the experiments was to develop and characterize a murine model for investigating the effects of prenatal cocaine exposure on the mother and fetus. Pregnant mice were separated into three groups: group 1 was treated with cocaine HCl at 10 mg/kg twice daily (COC); group 2 was treated with saline at 10 ml/kg twice daily (SAL); and group 3 was pair-fed with the COC dams and was injected with saline following the same schedule (SPF) from embryonic day (E) 8 to 17. We utilized high-pressure liquid chromatography (HPLC) with UV detector and electrochemical detector to test the concentrations of cocaine, dopamine and serotonin, as well as HE staining to observe morphological alterations of liver and placenta. Though less food intake and lower weight gain were observed in COC and SPF groups but not in SAL dams, lower fetal body weight and brain weight were only seen in COC offspring. Pharmacological analysis revealed that cocaine was found in fetal plasma at 15 min following intraperitoneal administration on E17, accompanied with elevated concentrations of dopamine (DA) and serotonin (5-HT) in fetal brain. We also observed morphological changes in liver and placenta of cocaine-exposed fetuses. The present study indicates that pregnancy cocaine exposure can lead to maternal undernutrition and developmental abnormality of the fetal brain, liver and placenta. It is suggested that the developmental abnormality of the fetuses induced by cocaine is due to the toxicological effect of cocaine but not to maternal undernutrition.
Animals ; Brain ; metabolism ; pathology ; Cocaine ; adverse effects ; blood ; Disease Models, Animal ; Dopamine ; metabolism ; Female ; Fetus ; drug effects ; pathology ; Liver ; pathology ; Malnutrition ; Maternal Exposure ; adverse effects ; Maternal Nutritional Physiological Phenomena ; Mice ; Mothers ; Placenta ; pathology ; Pregnancy ; Serotonin ; metabolism

Objective To screen down regulated genes and find down regulated novel genes in gastric cancer. Methods Genes mRNA expression were detected between gastric cancer and normal gastric mucous membrane of five patients using cDNA microarray. Genes mRNA expression signals on hybridization membranes were analysized with computer software. Down regulated genes in gastric cancer were screened. cDNA suppression subtraction library was established by counterpart normal gastric mucous membrane mRNA(Tester) subtracting gastric cancer tissues mRNA(Driver) of five patients. After identification of the subtraction library, positive clones choosen randomly were sequenced , and down regulated novel genes in gastric cancer were screened. Some of the genes were identified by RT PCR.Results Down regulated genes in gastric cancer consist of 60 genes including tumor suppressing genes, apoptosis related genes, DNA replication and transcript or translate related genes, cell cycle related genes, cell migration related genes, etc. Two unknown gene fragments in gastric cancer were cloned. Conclusions Sixty down regulated genes in gastric cancer are confirmed. They are involved in gastric tumorigenicity and metastasis. cDNA subtraction library of gastric cancer was constructed successfully. Two down regulated novel gene fragments in gastric cancer was found.

Objective To observe the effect of type 2 diabetes mellitus on serum platelet-derived endothelial cell growth factor (PD-ECGF) and vascular endothelial growth factor (YEGF) in patients with acute cerebral infarction.Methods Seventy-three patients with acute cerebral infarction (32 patients with non-concomitant diabetes were assigned to group A,and 41 patients with diabetes were assigned to group B),30 cases of type 2 diabetes mellitus patients with non-cerebrovascular disease hospitalized in the same period (group C),and 30 patients without cerebrovascular disease and diabetes mellitus (group D) were the subjects.The serum levels of PD-ECGF and VEGF in group A and group B were measured at 1,3,7,10 to 14 days after onset by double antibody sandwich enzyme-linked immunosorbent assay,which in group C and group D were measured at 1 st day..NIHSS scores were performed on group A and group B using the NIHSS Score (NIHSS) scale.Results The levels of PD-ECGF in group A were (5.93 ± 1.25),(5.93 ± 1.25),(4.19 ± 1.23),(3.67 ± 1.06) μg · mL-1,at 1,3,7,10 to 14 days after onset,The levels of PD-ECGF in group B were (2.88 ±0.54),(2.84 ±0.53),(2.81 ±0.41) and (2.86 ± 0.49) μg · mL-1.The VEGF levels in group A were (172.32 ± 31.91),(158.91 ± 31.84),(158.69 ± 29.27),(156.92±38.16),(159.64 ±27.21) and (159.91 ±40.25) pg· mL-1,The levels of VEGF in group B were (154.91 ±31.84),(158.69 ±29.27),(156.92 ±38.16),(159.64 ±27.21) pg · mL-1,the differences were statistically significant (all P ＜ 0.05).On the first day of admission,the PD-ECGF of group C and D were (2.25±0.49),(2.79±0.51) μg·mL-1,the VEGF were (94.90±19.85),(151.11±130.33) pg· mL-1,the differences were statistically significant (all P ＜ 0.05).The scores of NIHSS in group A and group B were (12.52 ±3.25) and (12.89 ±2.56).The difference was not statistically significant (P ＞ 0.05).On the 10 th to 14th day after onset,the NIHSS scores of group A and group B were (6.48 ± 2.15) and (4.24 ± 1.87) respectively,the difference was statistically significant (P ＜ 0.05).Conclusion Type 2 diabetes can reduce the expression of PD-ECGF and VEGF in patients with cerebral infarction and affect the prognosis of patients with type 2 diabetes mellitus.

BACKGROUNDBlocking the 4-1BB/4-1BB ligand (4-1BBL) signal may modulate the secretion of Th1/Th2 cytokines and prolong the survival of the grafts, which play a key role in organ transplantation tolerance. The aim of this study was to investigate the role of blockade of the 4-1BB/4-1BBL co-stimulatory pathway with 4-1BBL monoclonal antibody (mAB) in acute rejection of rat orthotopic liver transplantation.

METHODSThe orthotopic liver transplantation model was set up, while male Lewis rats were used as liver donors and Brown-Norway rats as recipients. The recipient rats were intravenously injected with anti 4-1BBL mAB or isotype control antibody. Groups were monitored for graft survival after transplantation. Plasma chemistry, including aspartate transaminase (AST), alanine aminotransferase (ALT), and bilirubin (BIL), was assayed. The concentrations of interleukin (IL)-2, IL-10 and interferon (IFN)-gamma in plasma were also measured by enzyme-linked immunosorbent assay. Allograft histology images were collected under light microscope and electron microscope.

RESULTSIsotype antibody treated recipients exhibited elevated plasma levels of liver injury markers including AST, ALT and BIL, progressive portal and venous inflammation and cellular infiltration of the liver allografts, and a mean graft survival time (MST) of 10.9 days. Administration of anti 4-1BBL mAB resulted in a decrease in plasma levels of liver injury markers and the concentrations of IL-2, IL-10 and IFN-gamma. The histological grade of rejection on day 7 decreased and MST (17.3 days) increased substantially.

CONCLUSIONSThese results demonstrate that attenuation of acute rejection follows the blockade of the 4-1BB/4-1BBL co-stimulatory pathway with 4-1BBL monoclonal antibody and strongly suggest it is a promising strategy to prevent progression of graft rejection by suppressing T cell-mediated immunity.

4-1BB Ligand ; immunology ; Alanine Transaminase ; metabolism ; Animals ; Antibodies, Monoclonal ; pharmacology ; therapeutic use ; Aspartate Aminotransferases ; metabolism ; Bilirubin ; metabolism ; Enzyme-Linked Immunosorbent Assay ; Graft Rejection ; immunology ; prevention & control ; Graft Survival ; drug effects ; Interferon-gamma ; blood ; Interleukin-10 ; blood ; Interleukin-2 ; blood ; Liver Transplantation ; adverse effects ; Male ; Rats ; Rats, Inbred Lew

BACKGROUNDThe antitumor role of Ras association domain family 1A (RASSF1A) gene and its potential molecular mechanisms are not well understood. The objective of this study was to observe the antitumor ability of RASSF1A in hepatocellular carcinoma, and study the mechanisms of cell apoptosis induced by RASSF1A.

METHODSAfter stably transfecting a RASSF1A (wild-type or mutant) expression vector into the BEL-7402 hepatocellular carcinoma cell line, RT-PCR and Western blotting was used to detect the RASSF1A expression levels in recombinant cells. The effects of wild-type RASSF1A on cell growth were observed in vitro by analyzing cell proliferation rate, cell colony formation, and in vivo by analyzing tumorigenesis in nude mice. In addition, the effect of RASSF1A gene expression on the chemosensitivity of human hepatocellular carcinoma cells to antitumor drugs was examined by inhibition of cell proliferation and the percentage of apoptotic cells.

RESULTSWild-type RASSF1A, not the mutant, suppressed cell growth in vitro and in vivo. Re-expression of wild-type RASSF1A could enhance the inhibition of cell proliferation and the percentage of apoptotic cells following cell treatment with mitomycin, but had no significant effect when combined with adriamycin, etoposide, 5-fluorouracil and cisplatin treatment.

CONCLUSIONWild-type RASSF1A inhibits cell growth and enhances cell chemosensitivity to mitomycin in hepatocellular carcinoma, suggesting that RASSF1A may serve as a new target for gene therapy in hepatocellular carcinoma patients.

Antineoplastic Agents ; pharmacology ; Apoptosis ; drug effects ; genetics ; Blotting, Western ; Carcinoma, Hepatocellular ; pathology ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Genetic Therapy ; methods ; Humans ; Mitomycin ; pharmacology ; Reverse Transcriptase Polymerase Chain Reaction ; Tumor Suppressor Proteins ; genetics ; metabolism ; physiology