OBJECTIVE: To explore the effect of traditional Chinese herbal medicine (TCM) for nourishing liver and kidney, clearing meridians and removing toxic substances, on the neurobehavioral manifestations and the activity of the dopamine D2 receptor in rat with levodopa-induced dyskinesias (LID). METHODS: The rat model of Parkinson's disease (PD) was established by injecting 6-hydroxydopamine (6-OHDA) into right substantia nigra of brain, then, the model of LID in rat was produced by injecting levodopa (LD) and benserazide for 4 weeks. The rats were divided into normal control group, 4-week LD treated group, 4-week LD plus TCM treated group, 8-week LD treated group, and 8-week LD plus TCM treated group, and the effect of the TCM on neurobehavioral manifestations was observed. The radioligand binding assay (RLBA) and Scatchard drawing were used to measure the maximal binding capacity of receptor (Bmax) and equilibrium dissociation constant (KD) of the dopamine D2 receptor in corpora striatum. RESULTS: Compared with the 4-week LD treated group and 8-week LD treated group, TCM could decrease abnormal involuntary movement scores of the rats with LID; the RLBA revealed that the dopamine D2 receptor Bmax significantly increased (P<0.05, P<0.01) and the KD significantly decreased (P<0.05). CONCLUSION: TCM can improve the activity of the dopamine D2 receptor and relieve the symptoms of LID.

Objective To observe the clinical efficacy of Zishen Pingchan Granules in treating non-motor symptoms (NMS) of Parkinson disease (PD). Methods A randomized, double-blind, placebo-controlled trial was used. 124 patients with PD were randomly divided into the treatment group (n=62) and the control group (n=62). Patients not yet taking Western medicine were suspended the application of Western medicine, receiving direct TCM treatment. Patients who had already taken Western medicine were added TCM on the basis of original medicine dosage and method. The treatment group was given Zishen Pingchan Granules, while the control group was treated with placebo granule, once a bag, twice a day. The treatment lasted for 12 weeks. The clinical efficacy was assessed by using unified PD rating scale (UPDRSⅡ–Ⅲ), the scale for outcomes in PD for autonomic symptoms (SCOPA-AUT), Parkinson disease sleep scale (PDSS) and dosage of levodopa. Results The UPDRSⅡ and UPDRS Ⅲscores of the treatment group and the control group after 4, 8, and 12 week of treatment had no difference at each time point compared with before treatment (P>0.05). There was no statistical significance in scores of UPDRSⅡ and UPDRSⅢ after treatment (P>0.05). Efficacy of SCOPA-AUT in treatment group after treatment was significantly better than the control group; There was statistical significance in constipation, salivation, nocturia and sweating in SCOPA-AUT between the two groups after treatment (P<0.05). Compared with before treatment, PDSS scores in treatment group after 8 and 12 weeks of treatment increased significantly (P<0.05). After 4, 8, and 12 weeks of treatment, PDSS scores in treatment group were much higher than control group (P<0.01). By comparing PDSS score in the two groups before and after treatment, it showed that treatment group had better efficacy in extending the time to sleep, reducing nocturia, nightmares, and hallucinations (P<0.05). After 12 weeks of treatment, dosage of levodopa in control group increased significantly, which treatment group was much lower than the control group (P<0.05). Conclusion Zishen Pingchan Granules can improve the autonomic dysfunctionin and sleep quality of PD patients, and can significantly reduce the dosage of levodopa.

Objective Construction of human IL‐4 recombinant expression vector and then conduct the expression ,purification and identification of human recombinant IL‐4 .Methods the open reading frame of IL‐4 was amplified by nest PCR with total RNA from PBMC of healthy volunteer .And then the amplified IL‐4 was inserted into pET101/D‐TOPO ,transformed into BL21 ,ex‐pressed ,purified and indentified .Results The size of amplified open reading frame of IL‐4 was about 460 bp and the sequence was correct .After transformed into BL21 ,the IL‐4 clone with higher expression level was selected by selection of different clones insert‐ed with IL‐4 and the size of expressed ,purified IL‐4 was about 28 × 103 .Western blot results showed that the size of single band was identical with the expected protein .Conclusion Human IL‐4 recombinant protein was got successfully .

Adult ; Bronchoalveolar Lavage ; methods ; Humans ; Male ; Middle Aged ; Pneumoconiosis ; therapy ; Quinuclidines ; therapeutic use

Objective:To single amino acid mutation of the full human scFvs against TSLP to enhance its affinity.Methods: The specific scFvs against TSLP was screened in our previous study and here the three-dimensional structures of TSLP and anti-TSLP scFvs were simulated by Discovery Studio system,then the molecular docking was made.The amino acids of binding epitope were randomly mutated and the mutated amino acids were selected which could remarkably improve the affinity of scFvs.The primers were designed based on the sequence of mutation amino acids and the scFv sequences were mutated by the overlapping extension PCR.The DNA of mutated scFvs was ligated with the expression vector pLZ16 and transformed into E.coli DH5αF′.Then the scFvs were expressed and the scFvs with improved affinity were selected by ELISA and BIAcore.Results: The five scFvs with single amino acid mutation were screened out by DS system,which could elevate the affinity of scFvs.The mutated anti-TSLP-scFvs were amplified by PCR,which size was about 1 000 bp.The mutated scFvs with correct sequence were expressed,and the mutated scFvs with improved affinity were detected by ELISA and BIAcore.The affinity of selected mutated scFv (M4) has been about 10 times higher than the scFv nonmutation.Conclusion: The affinity of anti-TSLP-scFv has been improved successfully.

Objective:Expression and purification of the α-HL of Staphylococcus aureus as antigen for making full human anti-α-HL antibody later ,providing of new treatment for Staphylococcus aureus infection.Methods:The total RNA of Staphylococcus aureus was extracted and the cDNA of α-HL was amplified by RT-PCR.The DNA of α-HL and pCold-TF plasmid was digested and ligated by T4 ligase and then transformed into E.coli TOPO 10.The recombinant plasmid α-HL/pCold-TF which verified by sequencing was trans-formed into E.coli BL21 for expression.The expression products was identified by SDS-PAGE and Western blot.Results: The size of amplified cDNA of α-HL was about 900 bp and the expressed soluble fusion protein of α-HL was about 90 kD(including the molecular chaperone in the vector ) after inducing expression for 24 h at 15℃.The Western blot results showed that the expressed protein was the fusion protein of α-HL.The purified α-HL was injected into BABL/c mice for making antiserum.The results showed that the antiserum had good binding activity with Staphylococcus aureus and the titer was greater than 10 000 times.Conclusion: The α-HL of Staphylococcus aureus was successfully cloned and the soluble fusion protein of α-HL was successfully expressed.

Interleukin family plays an important role in cleating the infection of Toxoplasma gondii in the body of host such as destroying the infected cells,constructing immunity system and regulating immunity balance,and so on.Here we reviewed the role of several important interleukin family members so as to provide scientific basis for the treatment of Toxoplasma gondii infection.

AIMTo prepare the micelles of stearic acid-grafted chitosan oligosaccharide and investigate the drug release from micelles.

METHODSMediated by a 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC), stearic acid (SA) was covalently attached to chitosan oligosaccharide (CSO), and the graft polymer (CSO-SA) was obtained. The critical aggregation concentration (CAC) of the CSO-SA was determined by measuring the fluorescence intensity of pyrene as a fluorescent probe. The effect of various pH dispersed media and concentration of tripolyphosphate sodium (TPP) on the micellar size distribution and zeta-potential measured by light scattering and electrophoretic mobility, was investigated. In buffers of different pH, the release profiles of methotrexate (MTX) from micelles were evaluated.

RESULTSThe CAC value of CSO-SA in deionized water was 0.05 g x L(-1). The mean diameter of CSO-SA micelles was 26.7 nm and the zeta potential was (55.9 +/- 0.1) mV. With the increase of TPP concentration, the size and MTX encapsulation of CSO-SA micelles increased, while the zeta-potential decreased. With the decrease of pH value of dispersed media, the size and zeta-potential of CSO-SA micelles increased, and the MTX encapsulation in CSO-SA micelles decreased. While the enhancement of drug release from the micelles was observed.

CONCLUSIONThe graft polymer of CSO-SA provides polymeric micelles, which possessed a low CAC value in aqueous media. The drug release in vitro from CSO-SA micelles was affected by the pH of delivery media.

Chitosan ; administration & dosage ; chemistry ; Drug Carriers ; Drug Delivery Systems ; Hydrogen-Ion Concentration ; Methotrexate ; administration & dosage ; chemistry ; Micelles ; Oligosaccharides ; administration & dosage ; chemistry ; Particle Size ; Polymers ; Polyphosphates ; Solubility ; Stearic Acids ; administration & dosage ; chemistry

Objective:To select specific and neutralizing scFv against IL-13 and soluble expression and identification them.Methods:In our previous study the scFv library were constructed,and here the scFv library was enriched and the positive scFv were screened from the enriched scFv library for three round.The specific positive scFvs with better affinity were ligated with expression vector for expression and identification.Results:After three rounds of enrichment,30%of clones were positive.The two specific scFvs with better affinity and neutralization were selected from almost 500 clones and then ligated with expression vector LZ16 for soluble ex-pression.The expressed scFvs were identified by western blot and biomolecular interaction analysis.Conclusion: The specific scFvs against IL-13 with better affinity and neutralization had been selected successfully.

Objective To find out the current situation of brucellosis infection and exposure status of family members of sheep farmers in the western pastoral areas of Jilin Province,and to provide a reference for control of human brucellosis spreading among family members.Methods On November 2012,Qianguo County was randomly selected using a multi-stage sampling method,and two townships,Chaganhua and Wulantala,were randomly selected in the county; half of the villages were selected from each township; all family members of the sheep farmers in these villages were investigated about their demographic characteristics (sex,age,education),high-risk behavior and information about brucellosis infection by using a questionnaire survey.Based on the principle of informed consent,respondents venous blood samples were collected.Brucellosis was confirmed with serum agglutination test (SAT).The effects of gender,age,education and other demographic data,high-risk behavior and high-risk behavior protection on the prevalence of brucellosis were studied.Results Out of the 403 copies of qualified questionnaires collected,84 people were found infected with brucellosis,and the infection rate was 20.84％ (84/403).Men infection [24.78％ (57/230)] was higher than that of women [15.61％ (27/173),x2 =5.038,P ＜ 0.05].The rates of eight kinds of high-risk behaviors were:helping feeding 86.85％ (350/403),cleaning sheepfold 80.40％ (324/403),holding lamp 71.71％ (289/403),delivering sheep 61.54％ (248/403),vaccinating sheep 53.85％ (217/403),apoblema 47.39％ (191/403),milking 22.08％ (89/403) and slaughtering sheep 14.89％ (60/403).The highest risk behavior was vaccinating sheep[24.40％(53/217)],and the lowest was milking [16.90％ (15/89)].The highest rate of basic protection was delivering a sheep [31.85％ (79/248)],the next was apoblema[27.23％ (52/191)],and the lowest was slaughtering sheep [8.33％ (5/60)].There was no statistical significant difference between brucellosis infection and the eight kinds of high-risk behaviors as well as the basic protective behaviors.Conclusions The prevalence of brucellosis infection among sheep farmers' family members is higher than others.Brucellosis infection-related contact manner is ubiquitous and the level of basic protection is low.We should carry out targeted health education on sheep farming household members to improve their level of protection,so as to reduce the risk of brucellosis infection.