Primary CNS lymphoma (PCNSL) is a rare malignancy with peculiar clinical and biologic features,aggressive course,and unsatisfactory outcome.the particular microenvironment of this malignancy,and sanctuary sites where tumor cells grow undisturbed, strongly affects treatment efficacy. The execution of prospective trials is also difficult because of the rarity of the tumor and the impaired general condition and poor performance status of patients. Chemotherapy is indispensable for the treatment of PCNSL. High-dose methotrexate is the most effctive drug.PCNSL is sensitive to irradiation,but responses to radiotherapy alone are short-lived. MTX-based chemotherapy followed by whole-brain radiotherapy prolonged survival but is associated with delayed neurotoxicity especially in patients older than 60 years. A new approach is attempted to treat PCNSL with chemotherapy alone with defered radiotherapy in old patients with similar survival rencenfly. High-dose chemotherapy with autologous stem cell transplantation HDC/ASCT yielded promising results for recurent PCNSL.Clinical investigation with Temozolomide or rituximab have been reported.Further studies with these new drugs are needed.

Trypanosoma is a human parasite severely affecting poor tropical areas.However,current frontline drugs for Trypanosoma treatment have severe side-effects with decreased effectiveness.Based on the fact that aminoacyl-tRNA synthetase is a bonafide drug target for several microorganisms,including bacteria and fungi,it is plausible that it may also be effective target of Trypanosoma.The Trypanosoma brucei leucyl-tRNA synthetase(tbLeuRS)was cloned,expressed and purified to develop an in vitro enzymatic assay system.The assay conditions were further optimized for the effective screening of tbLeuRS inhibitors thus establishing an anti-Trypanosoma drug screening system targeting tbLeuRS.The results indicated that this system can be employed for the effective screening of anti-Trypanosoma drugs with satisfactory specificity.In addition,this system can also be used for compound optimization,as well as IC50 testing.Using this system a series of compounds are identified that are effective Trypanosoma inhibitors without toxicity to human cells.Therefore,targeting tbLeuRS may represent a new venue for the development of anti-Trypanosoma drugs.

OBJECTIVETo investigate the effect of augmentative locking compression plate combined with bone graft in treating aseptic femoral shaft nonunion after intramedullary nailing.

METHODSTwenty-one cases with aseptic femoral shaft nonunion after intramedullary nailing from January 2007 to January 2013 were treated,including 18 males and 3 females with a mean age of 37.7 years (ranged from 23 to 64 years). The mean period of nonunion after surgery was 23.9 months (ranged from 9 to 62 months). According to Weber-Cech classification,10 of those 21 cases were hypertrophic nonunion,7 were atrophic, and 4 had oligotrophic fracture nonunion. All patients retained the original intramedullary nail, and applied with augmentation plating of 6 to 8 holes locking compression plate, unicortical fixation with 2 to 3 locking screws in the proximal or distal end, with simultaneous autologous iliac bone grafting. After treatment,all patients were allowed to partial weight-bearing until full weight-bearing according to the radiological results. All patients were followed up and were evaluated with clinical and imaging results.

RESULTSAll patients were followed up from 8 to 24 months, averaged (13.5±3.5) months,which showed clinical union at 4 to 8 months, averaged (6.0±1.0) months and radiological solid union at 7 to 12 months, averaged (9.1±1.5) months. No such complications as infection,hardware loosening or breaking were found.

CONCLUSIONAugmentative locking compression plate(LCP) combined with bone graft for aseptic femoral shaft nonunion after intramedullary nail has a satisfied clinical efficacy. It's an useful and simple method.

Adult ; Bone Nails ; adverse effects ; Bone Plates ; Bone Transplantation ; Female ; Femoral Fractures ; complications ; surgery ; Follow-Up Studies ; Fracture Fixation, Intramedullary ; adverse effects ; Fractures, Ununited ; complications ; surgery ; Humans ; Male ; Middle Aged ; Postoperative Complications ; etiology ; surgery ; Treatment Outcome ; Young Adult

This study is to establish physiologically based pharmacokinetic (PBPK) models of famitinib in rat and monkey, and then to predict the pharmacokinetics and tissue distribution of famitinib in human based on the PBPK models. According to published paper, previous studies and the chemical properties of famitinib predicted by ACD/ADME suite and SimCYP, the PBPK models of rat and monkey were established and optimized using GastroPlus. And then, the PBPK models were applied to predict the pharmacokinetic and tissue distribution of famitinib in human. The results showed that the PBPK models of rat and monkey can fit the observed data well, and the AUC0-∞, ratios of observed and calculated data in rat and monkey were 1.00 and 0.97, respectively. The AUC0-∞, ratios of observed and predicted data in human were 1.63 (rat to human) and 1.57 (monkey to human), respectively. The rat and monkey PBPK models of famitinib were well established, and the PBPK models were applied in predicting pharmacokinetic of famitinib in human successfully. Hence, the PBPK model of famitinib in human could be applied in future drug-drug interaction study.
Animals ; Antineoplastic Agents ; pharmacokinetics ; Haplorhini ; Humans ; Indoles ; pharmacokinetics ; Models, Biological ; Pyrroles ; pharmacokinetics ; Rats ; Receptor Protein-Tyrosine Kinases ; antagonists & inhibitors ; pharmacokinetics ; Tissue Distribution

This study was aimed to clone human intercellular adhesion molecule-1 (ICAM-1) gene, to transfect the constructed eukaryotic expression vector ICAM-1-GFP into CHO cells, as well as to detect ICAM-1-GFP expression in CHO cells binding with Molt-4 cells. ICAM-1 cDNA gene was amplified by RT-PCR and inserted in PMD(18)-T vector. Then ICAM-1 cDNA from pMD18-ICAM-1 vector was subcloned into eukaryotic expression vector pEGFP-C1 to construct recombinant ICAM-1-pEGFP-C1 vector. Restriction analysis and DNA sequencing were used to confirm the recombinant vector. After stable transfection of CHO-K1 cells with the recombinant vector, the expression and subcellular localization of ICAM-1-GFP were detected by RT-PCR, flow cytometry and fluorescence microscopy. The function of ICAM-1-GFP fusion protein was assessed by the binding of ICAM-1-GFP/CHO cells to Molt-4 cells. The results showed that 1622 bp full-length ICAM-1 cDNA obtained and was successfully ligated with pMD(18)-T-vector, subcloned to construct recombinant ICAM-1-pEGFP-C1 vector. Restriction analysis and DNA sequencing indicated that recombinant ICAM-1-GFP was successfully constructed and ICAM-1-GFP was expressed stably in CHO cells. ICAM-1-GFP expression was only observed in the cytoplasm of ICAM-1-GFP/CHO cells by fluorescence microscopy. The ICAM-1-GFP/CHO cells were bound to PMA-treated Molt-4 cells. The expression of MEM-148 was very weak in PMA-treated Molt-4 cells. It is concluded that the ICAM-1-GFP eukaryotic expression vector has been constructed successfully and expresses stably in CHO cells. PMA can increase the binding of Molt-4 cells to ICAM-1-GFP/CHO cells by inducing specialized form of ICAM-1 clustering.
Animals ; CHO Cells ; Cricetinae ; Cricetulus ; DNA, Complementary ; genetics ; Genetic Vectors ; Green Fluorescent Proteins ; genetics ; Humans ; Intercellular Adhesion Molecule-1 ; genetics ; Recombinant Proteins ; genetics ; Transfection

Objective To examine the relationship between CCT 5 ,γδ T cell and autoimmune diseases .Methods Recombinant CCT5 protein was cloned , expressed and purified in E.coli.Three peptides of CCT5 protein were used to prepare for anti-CCT5 monoclonal antibodies .Purified CCT5 protein was used to expand γδT cells from pe-ripheral blood mononuclear cells (PBMC).Plasma level of CCT5 in healthy donors, patients with rheumatoid arthritis (RA) and systemic lupus erythematosus (SLE) were detected by ELISA assays .The correlation analysis between plasma CCT5 concentration and the percentage of different subtypes of γδT cells measured by flow cytometry was made . Results The CCT5 gene was amplified by PCR and the length of the target fragment was 1 750 bp.The expressed 65 ku CCT5 protein was purified and validated by SDS-PAGE.Two paired monoclonal anti-CCT5 antibodies were screened to detect CCT5 protein in plasma.Immobilized recombinant CCT5 protein was able to induce specific sig-nificant amplification of peripheral γδT cells.Correlation analysis of 10 healthy donors indicated significant corre-lation between the plasma CCT 5 concentration and the proportion of Vγ9 and Vδ2 γδ T cells.The plasma CCT5 concentration significantly decreased in autoimmune diseases patients , including RA and SLE .Conclusions These data suggest that CCT 5 could be a novel Vγ9δ2 γδT cell-related factor in autoimmune diseases , which deepen the understanding of Vγ9δ2 γδT cell function in autoimmune diseases .

Objective To report a three-generation Chinese family with freckle and to make a genetic linkage analysis in this family.MethodsGenetic linkage analysis was carried out in this family using microsatellite markers distributed over chromosome 4q and 1.Two-point logarithm of odds(LOD)scores were calculated using the Linkage program package(version 5.1),and haplotype was analyzed with Cyrillic version 2.01 software.Results Freckle was inherited in an autosomal dominant pattern with a penetrance of99.9% in this family;linkage to chromosome 4q was ruled out however,supportive evidence was obtained for linkage to microsatellite markers D1S2635 and D1S2844 in chromosome 1q with a maximum LOD score of 1.50.Haplotype analysis in this family localized the locus of freckle to a 12 Mb region flanked by D1S2624 and D1S2799.Conclusions Freckle is a genetically heterogeneous disorder.The causative gene may be located in a 21.2 cM region on chromosome 1q22-24.

Adenocarcinoma ; diagnosis ; Biomarkers, Tumor ; metabolism ; Carcinosarcoma ; diagnosis ; Gastrointestinal Stromal Tumors ; metabolism ; pathology ; surgery ; Humans ; Male ; Middle Aged ; Neoplasm Metastasis ; Prostate ; pathology ; Prostatic Neoplasms ; metabolism ; pathology ; surgery ; Stromal Cells ; pathology ; Treatment Outcome

BACKGROUND:Both salvia miltiorrhiza injection and adipose-derived stem cells have evident neuroprotection against cerebral infarction in rats.OBJECTIVE:To observe the effect of salvia miltiorrhiza injection combined with adipose-derived stem cell transplantation on cerebral infarction in rats and the levels of cyclooxygenase-2 (COX-2) and beta-amyloid protein (Aβ) in the serum and brain tissues.METHODS:A rat model of cerebral infarction was successfully established in 63 rats,and then the model rats were randomly divided into three groups (n=21 per group):model group (cerebral infarction group with 30 μL of PBS via the tail vein),stem cell group (transplantation of adipose-derived stem cells,30 μL,3x106/L,via the tail vein),salvia miltiorrhiza injection + adipose-derived stem cell transplantation(combined group with transplantation of adipose-derived stem cells,30 μL,3x106/L,via the tail vein and intraperitoneal injection of salvia miltiorrhiza injection,0.5g/(kg·d) for consecutive 7 days).The modified neurological severity scores of each group were evaluated before and at 1,2,3,and 4 weeks after treatment.At 3 weeks after treatment,MTT assay was used to detect the serum levels of COX-2 and Aβ;2,3,5-triphenyltetrazolium chloride staining was performed to detect infarction size;and RT-PCR and western blot assay were used to detect the mRNA and protein levels of COX-2 and Aβ in the brain tissues.RESULTS AND CONCLUSION:The modified neurological severity scores in the three groups were ranked as follows:combined group ＜ stem cell group ＜ model group,and there were significant differences between groups (P ＜ 0.05).Compared with the model group,the levels of COX-2 and Aβ in the serum and brain tissues at protein and mRNA levels were significantly lower in the stem cell group and combined group;compared with the stem cell group (P ＜ 0.05),the levels of COX-2 and Aβ were significantly lower in the combined group (P ＜ 0.05).The infarct size was smallest in the combined group,followed by the stem cell group,and biggest in the model group (P ＜ 0.05).To conclude,salvia miltiorrhiza injection combined with adipose-derived stem cell transplantation can improve the neurological function of rats after cerebral infarction,probably through reducing the levels of COX-2 and Aβ in the rat brain and serum.

BACKGROUND: The expression of exogenous genes in mesenchymal stem cells is beneficial to enhance the transplantation effect and improve the differentiation rate. OBJECTIVE: To verify whether the transplantation of amniotic mesenchymal stem cells (AMSCs) can improve the renal function and blood coagulation in rats with nephrotic syndrome by means of vascular endothelial growth factor (VEGF) modification. METHODS: The 18 of 72 male Sprague-Dawley rats were assigned to normal group (without any treatment), and the other 54 rats were intravenously injected with doxorubicin to establish adriamycin nephropathy rat models which were randomly divided into three groups:model group (tail vein injection of 10 μL of 0.9% PBS injection), AMSCs group (tail vein injection of 10 Ml of AMSCs suspension), VEGF modified group (tail vein injection of 10 μL of VEGF modified AMSCs suspension). The injection in each group began at 24 hours after modeling, once a day for 3 consecutive days. The levels of total cholesterol (TC), triglyceride (TG), total protein (TP), albumin (Alb), high density lipoprotein (HDL), low density lipoprotein (LDL), creatinine (Cr), blood urea nitrogen (BUN) and coagulation index were measured by an automatic biochemical analyzer. The 24-hour urinary protein was determined by trichloroacetic acid method. The mRNA and protein expression of heparanase (HPA) and VEGF in renal tissue were detected by RT-PCR and western blot, respectively. RESULTS AND CONCLUSION: (1) Compared with the normal group, the contents of 24-hour urinary protein, TC, TG, LDL, Cr, BUN in the model group were significantly increased (P < 0.05), while the contents of TP, Alb, HDL were significantly decreased (P < 0.01). The serum levels of TC, TG, LDL, Cr, BUN in the AMSCs group and VEGF-modified group were significantly lower than those in the model group (P <0.05), while TP, Alb and HDL were significantly higher in the model group (P < 0.05). Compared with the AMSCs group, these indexes were significantly improved in the VEGF-modified group (P < 0.05). (2) The coagulation indexes of the model group showed a tendency of hypercoagulability. Compared with the model group, the hypercoagulability in the AMSCs and the VEGF-modified group were improved, especially in the VEGF-modified group. (3) The expression of HPA gene and protein in the model group was significantly higher than that in the normal group. The expression of HPA gene and protein in the AMSCs group and VEGF-modified group was lower than that in the model group. The expression of HPA gene and protein in the VEGF modified group was the lowest, while the expression of VEGF gene and protein in the VEGF modified group was the highest among the groups (P < 0.05), and there were significant differences between groups (P < 0.05). To conclude, VEGF gene-modified AMSCs can be transplanted via tail vein to improve renal function and hypercoagulable state in rats with nephrotic syndrome.