Acute Disease ; Adolescent ; Child ; Child, Preschool ; Drug Resistance, Multiple ; genetics ; Drug Resistance, Neoplasm ; genetics ; Female ; Gene Expression Regulation, Neoplastic ; Humans ; Leukemia ; physiopathology ; Male ; Neoplasm Proteins ; genetics ; RNA, Messenger ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Vault Ribonucleoprotein Particles ; genetics

Objective To identify the polyreactivity of a monoclonal natural anti-keratin autoantibody 3B4 and to analyze its possible molecular me chanism.Methods enzyme-linked immunosorbent assay (ELISA)and immunohistoche mistry were applied to test the binding reactivity of 3B4 against different anti gens and tissues.The variable region genes and their amino acid composition wer e sequenced.Results 3B4 could reacted with a range of antigens and tissues,i n addition to keratin and skin.The variable region genes of its light chain and heavy chain showed high homology with germline genes VK1 am4 and VH1 J558.42.H CDR3 region,which mainly composed of short side chain amino acids(from 294 to 324 nucleotides around the heavy chain),was the only motif that differs from ot her highly homologous immunoglobulin genes.Conclusions The monoclonal natural anti-keratin autoantibody 3B4,with its variable region genes highly homologo us to germline genes,is highly polyreactive.The flexibility of HCDR3 may contr ibute to the polyreactivity.

The present study sought to investigate the anti-inflammation and immunoloregulation effect of 17 Guizhi Fuling capsule ingredients. The anti-inflammatory ingredients on LPS-induced RAW264. 7 cell injury were assessed with ELISA and immunofluorescence. The release of IL-1β, TNF-α, PGE2 were detected with ELISA and the expression of COX-2 was detected with immunofluorescence. The effects of them on promoting splenic lymphocyte proliferation were assessed with MTT and Hoechst 33342 staining method. The results showed that 15 ingredients had obviously anti-inflammatory activity on LPS- induced injury and play the immunoloregulation roles. This study suggested that the 15 ingredients may be the active ingredients on pelvic infection.
Animals ; Anti-Inflammatory Agents ; pharmacology ; Capsules ; pharmacology ; Cyclooxygenase 2 ; immunology ; Drugs, Chinese Herbal ; administration & dosage ; Immunologic Factors ; pharmacology ; Inflammation ; drug therapy ; Interleukin-1beta ; immunology ; Macrophages ; drug effects ; enzymology ; immunology ; Male ; Mice ; Mice, Inbred BALB C ; Spleen ; cytology ; drug effects ; immunology ; Tumor Necrosis Factor-alpha ; immunology

Objective To observe the effect of allogenic bone marrow mononuclear cells(BM-MNCs) transplantation on myocardial apoptosis after acute myocardial infarction(AMI) in rats.Methods 40 Wistar rats were randomly divided into control group(n=20) and transplantation group(n=20).Myocardium around the infarcted left ventricular area of the rats in transplantation group were injected with BM-MNCs suspension beneath the epicardium.Myocardium the area of control group was injected with culture solution.Results After 4 weeks of the operation,the myocardial apoptosis index,the TNF-? content and the PDCD5 mRNA of transplantation group were all notably less than those of control group(P

The fibrous roots are the residues of production of cut crude drug of Danshen (Salvia miltiorrhiza). Enzymatic pretreatment and ultrasonic extraction are beneficial to extract effective constituents from fibrous roots more effectively. The present research was to optimize the enzymatic parameters by the central composite design and response surface method. Under the best conditions, the yields of total tanshinones and total salvianolic acids in the extracts of enzymatic pretreatment increased by 113.92% and 30.64%, comparing with the non-enzymatic extraction, respectively. TLC analysis also showed that the types of effective constituents in the two samples were not affected by enzymatic hydrolysis. Meanwhile, the complex correlation coefficients of the mathematical models were high, which provided a good prediction.
Diterpenes, Abietane ; Drugs, Chinese Herbal ; chemistry ; Phenanthrenes ; chemistry ; Plant Roots ; chemistry ; Salvia miltiorrhiza ; chemistry

Objective To investigate the effects of different doses of pituitary adenylate cyclase- activating polypeptide(PACAP)on the functional and morphological outcome in a mice model of Parkinson' s disease(PD)rendered by 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine(MPTP).Methods Male mice were treated with PACAP 0.02, 0.20 or 2.00 ?g by iv bolus for 7 days after MPTP was administered, and were compared with the saline-treated mice.The immunohistochemistry and Western blot were used to detect the alterations of PD biomarker including tyrosine hydroxylase(TH), dopamine transporter(DAT)and vesicular monoamine transporter2(VAMT2).In addition, monoamine neurotransmitters in the striatum of mice were measured by the high performance liquid chromatography (HPLC).Results TH immunohistochemistry indicated that the number of TH-positive neurons in the substantia nigra was increased in all PACAP-treated mice(PACAP(0.02 ?g/d)group was 93.33?4.87, F=85.85,P

Objective To explore the conditions for high expression of anti-HBsAg scFv A-15 in E. coli, increase the production of the scFv in the culture medium. Methods By changing induction occasion, concentration of inductor IPTG and induction time, influence of various conditions on expression of anti-HBsAg scFv A-15 was analyzed through ELISA. In addition, the effects of sucrose, glycine and Triton X-100 at different concentrations on the scFv excretion into culture medium was evaluation. Results The optimal expression conditions were as follows: the induction was started after culturing for 4 h, the concentration of IPTG was 0.5 mmol/L,and the induction lasted for 8 h .The scFv affinity in culture medium with 0.3 mol/L sucrose,2% glycine, 1% Triton X-100,16.78-fold higher, respectively than that without the three chemicals. The final yield of anti-HBsAg scFv A-15 was estimated to be 7.4 mg/L. Conclusion The conditions for production of anti-HBsAg scFv A-15 were optimized,which provides a practical method for more efficient production of the scFv in E. coli for further studying structure and function.

Objective To explore the antivirus function in vitro of earthworm coelomic fluid(ECF) by researching its effect on inhibiting respiratory syncytial virus(RSV).Methods By the method of Hep-2 cell culture and using ribavirin as a positive control,the anti-RSV effect of ECF was investigated by observing cytopathic effect (CPE) with MTT colorimetric assay.Results In Hep-2 cells,the CC50 of ECF and ribavirin were 3.11 mg/ml and 1.35 mg/ml separately.In the experiment of ECF directly killing RSV,the IC50 of ECF was 184.1 μg/ml,SI was 16.87;In the experiment of ECF preventing RSV invasion,no antiviral function of ECF within the experimental concentration range was observed;In the experiment of ECF inhibiting RSV replication,the IC50 of ECF was 1555.8 μg/ml,SI was 1.99.Conclusion ECF couldn't prevent virus from invading into host cell,but showed direct killing-virus function and inhibition of the virus replication.

OBJECTIVETo optimize a pretreatment method of urine proteomics in children with primary nephrotic syndrome.

METHODSUrine from children with primary nephrotic syndrome was treated in different pH and isolated by cold acetone precipitation for different durations. Then the amounts and kinds of proteins were compared by quantify, SDS-polyacrylamide gel electrophoresis (SDS-PAGE) and two-dimensional electrophoresis (2-DE) in order to optimize a way to deal with urine protein.

RESULTSMost proteins were obtained at pH 2.7. The amounts of protein precipitated by acetone for 0.5 hr was obviously less than those precipitated for 1 and 2 hrs (P<0.05), while there was no significant difference between the amount of protein precipitated for 1 and for 2 hrs. Protein precipitated by cold acetone for 1 hr at pH 2.7 was selected as the best pretreatment method. Satisfactory 2-DE maps can be acquired.

CONCLUSIONSUrine protein can be best obtained at pH 2.7 and precipitated by cold acetone for 1 hr.

Child ; Electrophoresis, Gel, Two-Dimensional ; Humans ; Hydrogen-Ion Concentration ; Nephrotic Syndrome ; urine ; Proteinuria ; urine ; Proteomics ; methods

Objective To observe the clinical efficacy of Tiao Shen Jie Jing needling (regulating the mind and release spasm) in treating post-stroke upper-limb spastic palsy. Method Ninety patients with post-stroke upper-limb spastic palsy were randomized into an observation group and a control group, 45 cases each. The observation group was intervened by Tiao Shen Jie Jing needling, while the control group was given ordinary acupuncture. The Modified Ashworth Scale (MAS), short-form Fugl-Meyer Assessment (SFMA) and Barthel Index (BI) were evaluated and recorded prior to the treatment, after 2-week and 4-week treatment and 3 months later after the intervention; the serumγ-aminobutyric acid (GABA) was detected before and after the treatment. Result The total effective rate was 83.3% in the observation group versus 61.0% in the control group, and the efficacy of the observation group was superior to that of the control group (P<0.05). The scores of each scale showed improvements after the treatment in both groups (P<0.05), and the improvements in the observation group were more significant (P < 0.05). The follow-up showed continuing improvements in the scales in the observation group, which were superior to those in the control group (P<0.05); after the treatment, the content of GABA in the observation group was significantly higher than that in the control group (P<0.05). Conclusion Tiao Shen Jie Jing needling can effectively ameliorate the upper-limb spasm,as well as the motor function of the upper extremities, improve the activities of daily living and maintain a long-term efficacy, better than ordinary acupuncture. The action mechanism is related to the regulation of GABA content in serum.