AIMTo clone the gene of staphylococcal enterotoxin C2 and express it in the form of a soluble fusion protein in E. coli. Then the activation of SEC2 on mice lymphocyte and its lethal effects on tumor cells were studied.

METHODSStaphylococcus aureus SEC2 gene was cloned into GST gene fusion vector pGEX-4T-1. The resultant plasmid pGEX-4T-SEC2 was used to transform E. coli BL21, where the GST-SEC2 fusion protein was expressed efficiently. The rSEC2 protein was purified with Glutathione Sepharose 4B affinity column and digested with thrombin. The in vitro culture system was utilized to observe the activation of the SEC2 on mice lymphocyte and the lethal effects on tumor cells of the activated mice lymphocyte.

RESULTSThe proper gene of SEC2 was cloned and purified rSEC2 was obtained. The MTT results indicated that rSEC2 have strong ability to stimulate mice lymphocyte to proliferate with a dose-dependent manner. With the proliferation of mice splenic lymphocyte, rSEC2 has a strong lethal effect on tumor cells B16, K562 and K562-AD.

CONCLUSIONIn this study, the gene of SEC2 was cloned and the rSEC2 protein was obtained, which had strong lethal effect on tumor cells B16, K562 and K562-AD.

Animals ; Cell Line, Tumor ; drug effects ; Cell Proliferation ; Cloning, Molecular ; Enterotoxins ; genetics ; metabolism ; pharmacology ; Escherichia coli ; genetics ; metabolism ; Female ; Genetic Vectors ; Glutathione Transferase ; genetics ; Lymphocyte Activation ; drug effects ; Lymphocytes ; cytology ; immunology ; Male ; Mice ; Mice, Inbred ICR ; Recombinant Fusion Proteins ; genetics ; metabolism ; pharmacology ; Spleen ; cytology ; Transfection

OBJECTIVETo evaluate the roles of plasma mast cell carboxypeptidase and chymase in the diagnosis of allergic diseases by measuring the contents of both in children.

METHODSA total of 59 children with allergic diseases and 53 healthy children were recruited into the study. Plasma levels of mast cell carboxypeptidase and chymase were measured using ELISA.

RESULTSThe plasma levels of mast cell carboxypeptidase and chymase in children with allergic children were 1.089 ± 0.752 ng/mL and 0.905(0.375-2.318) ng/mL, respectively, which were significantly higher than those in healthy children [0.593 ± 0.380 ng/mL and 0.454 (0.097-1.077) ng/mL respectively; P<0.05]. There was a significantly positive correlation between plasma mast cell carboxypeptidase and chymase levels in children with allergic diseases (r=0.684, P<0.01).

CONCLUSIONSPlasma levels of mast cell carboxypeptidase and chymase increase in children with allergic diseases, suggesting that mast cell carboxypeptidase and chymase may serve as the indexes for the diagnosis of allergic diseases.

Adolescent ; Carboxypeptidases ; blood ; Child ; Child, Preschool ; Chymases ; blood ; Female ; Humans ; Hypersensitivity ; diagnosis ; enzymology ; Infant ; Infant, Newborn ; Male ; Mast Cells ; enzymology

OBJECTIVETo investigate the effects of microencapsulated Chinese hamster ovary (CHO) cells modified with maspin gene on the motility and adhesiveness of breast carcinoma cells Bcap37 and to explore the possibility and feasibility of its clinical application in treatment of malignant tumors.

METHODSAfter the Bcap37 cells were co-cultured with the microencapsulated CHO cells modified with maspin gene, their motility and adhesion to vascular endothelial cells (ECV304), changes in CD44v6 and E-cadherin expression were examined.

RESULTSAfter the treatment, the motility of Bcap37 cells, their adhesion to vascular endothelial cells ECV304 and the CD44v6 expression were significantly reduced. The adhesiveness of Bcap37 cells and their E-cadherin expression were significantly enhanced.

CONCLUSIONThe microencapsulated CHO cells modified with maspin gene decrease motility and adhesiveness of breast carcinoma cells Bcap37, which help explain the anti-metastatic effects of maspin.

Animals ; Breast Neoplasms ; pathology ; CHO Cells ; Capsules ; Cell Adhesion ; Cell Movement ; Coculture Techniques ; Cricetinae ; Cricetulus ; Female ; Glycoproteins ; biosynthesis ; genetics ; Humans ; Hyaluronan Receptors ; biosynthesis ; genetics ; Serpins ; genetics ; Tumor Cells, Cultured

OBJECTIVETo investigate the association between the C311S polymorphism of paraoxonase 2 (PON2) gene and ischemic stroke in Chinese type 2 diabetes mellitus (T2DM) patients.

METHODSA case-control study of 279 Chinese subjects (including 162 T2DM with or without ischemic stroke and 117 non-diabetic control) was performed. Genotype frequencies of C311S polymorphism were studied by polymerase chain reaction-restriction fragment length polymorphism(PCR-RFLP) analysis with DdeI digestion.

RESULTSC311S polymorphism of PON2 gene was detected in Chinese with the C/S allele frequencies 0.145 and 0.855. The frequency distribution showed significant difference between Chinese and Asian Indian. Furthermore, the genotype distribution (SS, CS and CC) of the PON2 C311S gene polymorphism exhibited a significant difference between T2DM patients complicated with ischemic stroke and T2DM without ischemic stroke, the former had a significantly higher C allele frequency(P<0.05).

CONCLUSIONThe above data indicate that the polymorphism at codon 311(Cys --> Ser)in the PON2 gene is associated with ischemic morbidity in Chinese T2DM patients and C allele might be a risk factor.

Adult ; Aryldialkylphosphatase ; genetics ; Asian Continental Ancestry Group ; genetics ; Diabetes Mellitus, Type 2 ; complications ; genetics ; Female ; Humans ; Male ; Middle Aged ; Polymorphism, Genetic ; Stroke ; etiology ; genetics

Female ; Humans ; Manipulation, Orthopedic ; Middle Aged ; Radius Fractures ; surgery ; Ulna Fractures ; surgery ; Ulnar Nerve Compression Syndromes ; surgery

OBJECTIVETo study the distribution and expression of fibromodulin, decorin and biglycan in developing normal periodontal tissues, so as to understand its role in periodontal tissue formation.

METHODSThirty six BALB/c mice in different developing stages were killed and their bilateral mandibular first molars with surrounding alveolar bones and gingival tissues were taken out, Power Vision two steps immunohistochemical method with anti-fibromodulin, anti-decorin and anti-biglycan was used to detect the tissue distribution and cellular localization of fibromodulin and related proteoglycans, decorin and biglycan.

RESULTSFibromodulin was strongly expressed in the subcutaneous gingival connective tissue, periodontal ligament, mainly in gingival and periodontal fibroblasts as well as their matrices. Strong expression was also noted in the area close to the interfaces of periodontal ligament-alveolar bone and periodontal ligament-cementum. Decorin was strongly expressed in the area of gingival connective tissue, periodontal ligament and the surface of alveolar bone, while biglycan was stained evidently in gingival connective tissue throughout the period of investigation, but negative in the surface of alveolar bone and osteoblasts.

CONCLUSIONSFibromodulin may interact with decorin and biglycan to regulate the network formation of gingival connective tissues and periodontal collagen fibers, and may be involved in mineralization of the alveolar bone and cementum.

Alveolar Process ; cytology ; growth & development ; Animals ; Biglycan ; Decorin ; Extracellular Matrix Proteins ; analysis ; Fibromodulin ; Gingiva ; chemistry ; growth & development ; Immunohistochemistry ; Mice ; Mice, Inbred BALB C ; Osteoblasts ; chemistry ; Periodontal Ligament ; chemistry ; growth & development ; Proteoglycans ; analysis ; Tooth Germ ; chemistry

OBJECTIVE Acetylated modification was applied to modify the structure of polysaccharide from stems and leav-es of Abelmoschus manihot,and the in vitro immunoregulatory activity of polysaccharides were evaluated,which was expected to provide the scientific evidence for the resource utilization of A.manihot disused parts.METHODS The crude polysaccha-ride(SLAMP)was obtained by water extraction and alcohol precipitation,and neutral polysaccharide(SLAMP-a)was further prepared by DEAE-52 ion exchange column.Acetic anhydride was used as acetylating agent to synthesize three acetylated de-rivatives (SLAMP-a1 ,SLAMP-a2 and SLAMP-a3).Their structures were preliminarily identified by chemical composition a-nalysis,pre-column derivatization HPLC method and IR spectrum.In addition,in vitro immunoregulatory activity of SLAMP-a and three acetylated derivatives was evaluated by splenocyte proliferation together with its effects on NO production of RAW264.7.RESULTS SLAMP-a showed little immunomodulatory activity and the total sugar content was 99.76%;Among the three derivatives,only SLAMP-a1 could significantly stimulate the proliferation of spleen cells and activate RAW264.7 to product NO.SLAMP-a1 mainly contained glucose with a few of mannose,galactose and arabinose,and the total sugar and DS was 82.50% and 0.62 respectively.CONCLUSION In vitro immunomodulatory activity of polysaccharide SLAMP-a from stems and leaves of A.manihot is significantly enhanced by acetylation and SLAMP-a1 possesses the potential to develop as immunoregulatory agents.The acetylation modification is an effective way for resource utilization of polysaccharide from the disused stems and leaves of A.manihot.

Objective To reveal the relationships between epileptic seizure in patients with tuberous sclerosis complex (TSC) and their intelligence.Methods A retrospective analysis was performed on the clinical data of 110 patients with seizure secondary to TSC,admitted to our hospitals from May 2007 to May 2012; the prognostic factors for mental retardation were analyzed.Results There were 67 males and 43 females in the TSC patients with seizures.The age of seizure onset ranged from 10 days to 28 years; the onset age was less than 1 year in 30％ and less than 3 years in 65％ of TSC patients.Twelve patients presented epileptic spasms; 40 had normal intelligence and 51 had mental retardation.In patients having onset age younger than 1 year,80％ had mental retardation; 92％ patients with epileptic spasms had mental retardation; 65％ patients with two seizure patterns had mental retardation; 62％ TSC patients with seizure frequency exceeding 4 times per month and 63％ patients with seizure history exceeding 5 years both had mental retardation.Conclusion The patients with seizure onset age less than 1 year old and having a history of epileptic spasms are more prone to mental retardation; so early control of seizure should be done to improve the intelligence.

Adolescent ; Adult ; Female ; Humans ; Male ; Middle Aged ; Surgical Flaps ; Thumb ; surgery ; Toes

Adolescent ; Adult ; Aged ; Child ; Endoscopes ; Female ; Humans ; Laser Therapy ; methods ; Male ; Microsurgery ; methods ; Middle Aged ; Prognosis ; Skull Base Neoplasms ; mortality ; pathology ; surgery