Aim To establish and evaluate Sprague-Dawley (SD) rats model with yang-deficiency and constipation at the direction of traditional Chinese medicine theories. Methods Rats model with yang-deficiency and constipation were induced by orally administration of white vinegar and active carbon-ice water. Changes of model rats in common biological characteristic, the number and time of excretion stool, colonic electromyography, gastrointestinal hormones and intestinal neurotransmitters were observed. Results Compared to the control group: changes of the model group in common biological characteristic including abdominal distension, puffed and withered hair, sluggish response, falling body weight, black and dry stool; Excretion time of the first granule stool became longer; Number of excretion stool within six hours became shorter; Intestine pushing rate decreased, and secretion of gastrointestinal hormones and intestinal neurotransmitters disordered, etc. After being treated with Tongbianling, the syndrome and indexes of model rats markedly improved. After 6 days′ resume, the model group in the experiment of excreting stool had markedly signification in comparison with the control group. Conclusion SD rats model with yang-deficiency and constipation could be stably established by administering white vinegar and active carbon-ice water.

A simple and rapid gas chromatography/mass spectrometry (GC/MS) analysis method was developed for the determination of fat-soluble parts of sinapis semina. Four major compounds were chosen as marker compounds to evaluate the method. Various extraction techniques were evaluated and the greatest efficiency was observed with sonication extraction using diethyl ether. The method was valuated as follows:acceptable apparatus suitability was obtained by testing the resolutions, tailing factors and theoretical plate number of the marker compounds; the precision and reproducibility, expressed as relative standard deviation (RSD), fell within the prescribed limits. Eight samples of sinapis semina collected from markets in Xi’an were monitored by using the method. The fingerprints of those samples were analyzed by hierarchical cluster analysis (HCA) similarity analysis. The result indicated that the combination of fingerprint and HCA could be used to analyze sinapis semina from different habitats.

Objectiye To study the mechanism by which transcranial magnetic stimulation (rTMS) affects cognitive dysfunction in vascular dementia (VD). Methods Thirty-six male Wistar rats were randomly divided into a control group, a VD group, a low frequency rTMS group and a high frequency rTMS group. Two-vessel occlusion was employed to induce VD models. Low frequency rTMS group rats were given 0.5 Hz rTMS for six weeks. High frequency rTMS group rats were given 5 Hz rTMS for six weeks. Morris' water maze test was used to measure their spatial learning ability and memory. The ultrastructures of the synapses in the four groups were detected with transmission electron microscopy. The expression of synaptophysin (SYN), brain derived neurotrophic factor (BDNF) and Nmethyl-D-aspartate receptor 1 ( NMDAR1 ) mRNA and protein in the hippocampus were determined by real-time PCR and Western blotting. Results The behavior and morphology of the rats treated with rTMS improved. The average expression of SYN, BDNF and NMDAR1 mRNA and protein in the low frequency rTMS group and the high frequency rTMS group were significantly higher than in the VD group. Conclusion rTMS can provide a rehabilitative effect for VD. The mechanism might be associated with enhancing the expression of SYN, BDNF and NMDAR1 in the hippocampus.

Objective To study the effects of repetitive transcranial magnetic stimulation(rTMS) of the supplementary motor area(SMA)on the cortical excitability in patients with Parkinson's disease(PD).Methods Sixteen patients with PD were included in this study.The motor evoked potentials(MEP)and the N30 component of somatosensory evoked potentials(SEP) were assessed for each patient before and after 1200 pulses of rTMS of the SMA at 5 Hz and an intensity of 100% of relaxed motor threshold (RMT) for the abductor pollicis brevis.Results Ten minutes after the rTMS intervention,the peak-to-peak amplitude of the SEP component P20-N30 increased significantly(P<0.05),with the P/F index decreased simultaneously(P<0.05).The MEP amplitude increased significantly,and reached the highest value at 10min after the rTMS intervention. Conclusion 5 Hz rTMS of the SMA can improve the excitability of the SMA itself temporarily.Meanwhile,it can induce a short-lasting facilitation of the excitability of M1 connected with SMA.

Objective To construct the eukaryotic vector that expressing hepatitis B virus (HBV) S and the single fragment of variety chain (ScFv) of monoclonal antiboy against cytotoxic Tlymphocyte-associated antigen-4 (CTLA-4) and to analyze the immunological activity of recombinant S-ScFv protein.Methods The oringially constructed pSect2/ScFv4F10 and pSect2/S were double enzyme digested by Sfi I and Hind Ⅲ,respectively.Then the HBV S gene was cloned into the pSect2/ScFv4F10 vector.The pSect2/ScFv4F10 and pSect2/S-ScFv4F10 were expressed in Chinese hamster ovary (CHO) cells,and the expressed proteins were verified through sodium dodecyl sulfatepolyacrylamide gelelectrophoresis(SDS-PAGE)andWesternblotting.Afterultrafiltration concentration and affinity chromatography,the biological affinity of the expressed ScFv4F10 and S-ScFv4F10 proteins were examined by competitive enzyme-linked immunosorbent assay (ELISA) and surface plasmon resonance (SPR) technology.The comparison between groups was done by One-way ANOVA.ResultsThe eukaryotic expression vector of pSect2/S-ScFv4F10was successfully constructed,and relative molecular mass of the expressed protein of S-ScFv4 F10 was about 52 000 that analyzed by SDS-PAGE and Western blotting.With the fixed concentration of 4F10-mAb against CTLA-4,the A570 value of the mixed reaction with purified CTLA-4 antigen gradually increased with the decrease of ScFc fusion protein proportion; when the molar ratio of ScFv,S-ScFv4F10∶4F10=2∶1,the competitiveinhibitionratesagainst 4F10conjugatedantigenwere72.6％and64.5％,respectively.The affinity constants of association kinetics for CTLA-4 mAb,ScFv4F10 and S-ScFv4F10 with CTLA-4 antigen were 7.29 × 108 mol/L,9.52 × 106 mol/L and 2.04 × 106 mol/L,respectively,and the dissociation constants of KD were 1.40 × 10-9 mol/L,1.05 × 10-7 mol/L and 4.91 × 10-7 mol/L,respectively.ConclusionsThe eukaryotic expression vector of pSect2/S-ScFv4F10is successfully constructed,and the recombinant protein of S-ScFv4 F10 has a fairly high affinity with CTLA-4 antigen.

OBJECTIVETo observe the clinical efficacy and safety on moderate and severe persistent allergic rhinitis treated with acupuncture.

METHODSSixty-six patients of moderate and severe persistent allergic rhinitis were randomized into an acupuncture group (34 cases) and a western medication group (32 cases). In the acupuncture, group, acupuncture was applied to Dazhui (GV 14), Feishu (BL 13), Pishu (BL 20), Ganshu (BL 18) and Shenshu (BL 23) in the prone, retained for 20 min; then in the supine, at Baihui (GV 20), Yintang (GV 29), yingxiang (LI20) Taichong (LR 3) and Hegu (LI 4), retained for 20 min. Acupuncture was given once every two days, three times a week, continuously for 8 weeks. In the western medication group, cetirizine hydrochloride was taken orally, 10 mg each time, once every day, continuously for 8 weeks. Separately, before treatment, after the treatment of 1 and 2 months and in 1 month after treatment, the total nasal symptom score (TNSS), the scores in the emotion rating scale for Ganzangxiang of TCM (ERSG) and the rhinoconjunctivitis quality of life questionnaire (RQLQ) were observed in the patients of the two groups. The clinical efficacy was compared between the two groups.

RESULTS(1) For TNSS, the results after 1 and 2 months treatment and in 1 month after treatment were all, reduced as compare with that before treatment separately in the two groups (P < 0.05, P < 0.01) The result after 2 months treatment was lower than that after 1 month treatment in the acupuncture group (P < 0.05). In 1 month after treatment, the result in the acupuncture group was lower than that in the western medication group (P < 0.05). (2) For ERSG, the score after 2 months treatment was lower than that before treatment in the two groups (both P < 0.05). The score after 2 months treatment in the acupuncture group was lower than that in the western medication group (P < 0.05). (3) For RQLQ, the score after 1 month treatment was lower than that before treatment and the score after 2 months treatment was lower than that after 1 month treatment in the two groups (all P < 0.05). The score after 1 and 2 months treatment and in 1 month after treatment in the acupuncture group was lower than that in the western medication group separately (all P < 0.05). (4) The total effective rate was 91.2% (31/34) in the acupuncture group and was 90.6% (29/32) in the western medication group, without significant difference between the two groups (P > 0.05).

CONCLUSIONAcupuncture is the safe and effective intervention on moderate and severe persistent allergic rhinitis. Compared with the western medicine group, the efficacy in the acupuncture group presents much more advantageous at its durability.

Acupuncture Points ; Acupuncture Therapy ; Adult ; Female ; Humans ; Male ; Middle Aged ; Quality of Life ; Rhinitis, Allergic ; diagnosis ; therapy ; Young Adult

Objective To investigate the induction and function of circulating CD4+ CD25+ regulatory T cells(CD4+CD25+ Treg cells)in patients with chronic hepatitis B,and their role in the progress of chronic HBV infection.Methods Twenty two patients with chronic hepatitis B(CHB),11 patients with acute hepatitis B(AHB)and 20 healthy blood donators were enrolled in the study.After stimulating with recombinant HBV antigen or anti-CD3 mAb,the fraction of CD4+ CD25+Treg cells in peripheral blood mononuclear cells(PBMC)from different subjects was analyzed using flow cytometry.CD4+CD25+ Treg cells and effector cells(PBMC-Treg)were sorted out and cultured alone or together in vitro,and the cell proliferation and cytokines production were measured. Serum HBV titers,HBV markers and liver function were also evaluated.Results The fraction of circulating CD4+CD25+ Treg cells induced in PBMC from CHB patients under the stimulation of recombinant HBsAg was much higher than that in AHB patients(F=3.47,P＜0.05)and normal controls(F=4.98,P＜0.05).With HBsAg stimulation,cell proliferation and cytokines production (IFN-γand IL-10)of CD4+ CD25+ Treg cells either from patients or from normal controls was very low when cultured alone,and there was no significant difference between two groups(P＞0.05).Cell proliferation and IFN-γproduction of effector cells from AHB patients were higher than those from CHB patients,however,when effector cells were cultured with CD4+Cd25+Treg cells at a ratio of 5:1,the cell proliferation and IFN-γ production were significantly inhibited (F=4.66,5.24,P＜0.05).Conclusion Circulating nificantly suppress the production and IFN-γ production of effector T Cells,which may be associated with the progress of chronic HBV infection.

Objective To explore the expression and function of miR-125b, miR-30b and miR-424 in endometrial cells. Methods Human endometrial samples were obtained in natural cycles and stimulating cycles. Endometrial epithelial cells (EECs) and endometrial stromal cells (ESCs) were isolated and confirmed by immunofluorescence. The expressions of miR-125b, miR-30b and miR-424 were detected by real-time PCR. Results The expression levels of miR-125b, miR-30b and miR-424 were higher in proliferative phase in ESCs than those in EECs. And in EECs, the expression levels of miR-125b, miR-30b and miR-424 were significantly up-regulated in secretory phase than in proliferative phase, while it was stable in ESCs. In addition, the expressions of miR-125b in EECs and miR-30b were increased in ESCs in women with elevated progesterone on the day of HCG administration than those of the control. The target genes of miR-125b, miR-30b and miR-424 mainly participated in cell migration and motion, cell-cell adherens junction and Wnt signaling pathway. Conclusion miR-125b, miR-30b and miR-424 were differently expressed in endometrial cells in different phases, and may participate in regulation of endometrial receptivity.

BACKGROUND:Upright rat intervertebral disc degeneration model can prove that intervertebral fusion can aggravate the adjacent-segment intervertebral disc degeneration, but cannot affirm that spinal non-fusion technology has more advantages. OBJECTIVE:Base on the affirmed intervertebral disc degeneration models of Beagle dog, we performed intervertebral fusion to affirm whether or not the intervertebral fusion can aggravate the adjacent-segment intervertebral disc degeneration. METHODS:Twelve Beagle dogs were randomly divided into two groups. The control group animals were stabbed percutaneous to injure L5/6 disc. The experimental group animals were stabbed percutaneous to injur L5/6 disc. One month later, we made L4/5 intervertebral fusion. At 3 and 6 months postoperatively, lumbar MRI was conducted. L5/6 discs were harvested and subjected to histological observation and PCR. RESULTS AND CONCLUSION:(1) MRI:in the experimental group, 3 and 6 months after fusion, different degrees of intervertebral disk herniation appeared. In the control group, no obvious intervertebral disk herniation appeared. (2) Histological observation:6 months after fusion, in the experimental group, intervertebral disc annulus and nucleus pulposus were disordered;there was a gap in the fold, and the number of nucleus pulposus cel s decreased. In the control group, nucleus pulposus and anulus fibrosus boundary was clear;col agen was loose and arranged regularly. Cel number was more. Anulus fibrosus was close to normal. Col agen fibers were regular and dense. In the experimental group, 3 and 6 months after fusion, type II col agen-positive cel s were less than the control group (P<0.05). Moreover, type II col agen-positive cel s were less at 6 months than at 3 months in the experimental group (P<0.05). (3) PCR:Bone morphogenetic protein 15 and tissue inhibitor of matrix metal oproteinase 1 gene expression was higher in the experimental group than in the control group at 3 and 6 months after fusion (P<0.05). Bone morphogenetic protein 15 and tissue inhibitor of matrix metal oproteinase 1 gene expression was higher in the experimental group at 6 months than that at 3 months (P<0.05). (4) These results suggest that intervertebral fusion can aggravate the adjacent-segment intervertebral disc degeneration.