Objective To investigate the effect of hypoxia-inducible factor-1? gene on the proliferation and differentiation of neural stem cells after focal cerebral ischemia in rats,and to explore the mechanism of the effect. Methods Middle cerebral artery occlusion(MCAO) and reperfusion models were established and divided into sham group,NS group,AD group and Ad-HIF-l? group.NS,AD and Ad-HIF-l? were injected into the ischemic ventricle respectively.The mNSS was evaluated the expression of EPO was observed,and the number of BrdU positive cells in subventricular zone and that of BrdU/NF200,BrdU/GFAP double labeled cells in cortex were calculated by immumofluorescence method.Results The mNSS were statistically different between Ad-HIF-l? group and the other three groups;the expression of EPO was higher in Ad-HIF-l? group;the number of BrdU positive cells increased obviously in Ad-HIF-l? group;the cellular rebirth and differentiation demonstrated that there existed a significant difference(P

OBJECTIVE: To observe the effect of long-snake moxibustion on pain, functional disorder and body constitution in the patients with lumbago of cold-dampness type. METHODS: A total of 90 patients with lumbago of cold-dampness type were randomized into long-snake moxibustion, acupuncture and medication groups, 30 cases in each group. In the long-snake moxibustion group, long-snake moxibustion was exerted on the spinal (between Dazhui[GV14] and Yaoshu[GV2]) with the self-prepared moxa powder formula combined with fresh ginger and moxa wool, once a week, for 4 weeks. Regular acupuncture was given to the acupuncture group, 30 min each time, once a day, 8 times as a treatment course, at the interval of 2 days between the courses. The duration of treatment was 1 month. In the medication group, Diclofenac Sodium was prescribed for oral administration, 25 mg each time, three times a day, and Mecobalamine tablets, 0.5 mg each time, 3 times a day. The duration of medication was 1 month. Before and after treatment, the differences were evaluated in the scores of visual analog scale (VAS), the present pain intensity(PPI), Oswestry disability index (ODI) and the scale of yang-deficiency body constitution among the 3 groups. RESULTS: After the treatment, VAS and PPI score and ODI were significantly reduced in the patients of the 3 groups as compared with those before treatment (all P<0.05). Compared with the medication group, ODI score was significantly reduced in the long-snake moxibustion group and the acupuncture group (P<0.05). After the treatment, the scores of the scale of yang-deficiency body constitution were significantly reduced in the long-snake moxibustion group and the acupuncture group (P<0.05) as compared with those before the treatment, but not statistically significant in the medication group in comparison before and after the treatment (P>0.05). The score of the scale of yang-deficiency body constitution in the long-snake moxibustion group was significantly lower than that in the acupuncture group and the medication group (P<0.05). Regarding the clinical effect, the effective rate was 93.33% (28/30) in the long-snake moxibustion group and it was 86.67% (26/30) in the acupuncture group and 73.33% (22/30) in the medication group after the treatment. The effective rate in the long-snake moxibustion group was significantly higher than those in the acupuncture group and the medication group (P<0.05). CONCLUSION: The long-snake moxibustion therapy can achieve significant effect on lumbago of cold-dampness type. This therapy can alleviate pain, relieve the functional disorders and improve the conditions of body constitution in the patients. Hence, it is applicable for the clinical promotion.

Objective: To test the feasibility of drag-reducing polymers (DRP) for improving coronary microcirculation in a canine model in order to provide the experimental basis for treating myocardial microcirculation dysfunction. Methods: A total of 8 dogs received open-chest surgery and they had intravenous injections, in turn, with adenosine (ADN), DRP 250 mg/L and DRP+ADN. The function y=A × (1-e-βt) was used to calculate the myocardium capillary volume (A value), capillary velocity (β value) and myocardial blood lfow (A ? β value) by myocardial contrast echocardiography. Results: With DRP infusion, the A value in experimental canine was similar to the baseline condition,P>0.05; while theβ value and A ? β value were signiifcantly increased as (0.57 ± 0.10) 1/s vs (0.23 ± 0.03) 1/s,P<0.01 and (11.51 ± 1.96) VI/s vs (5.15 ± 0.86) VI/s,P <0.05 respectively. With combined infusion of DRP+ADN, the β value and A ? β value were similar to the baseline condition, bothP>0.05. Conclusion: DRP improved coronary microcirculation primarily by modulating the β value in experimental canine model, and hopefully, this unique hemodynamics could provide a new approach for treating myocardial microcirculation dysfunction.

OBJECTIVETo observe the effects of electroacupuncture (EA) at "Sanyinjiao" (SP 6) on urodynamics indices in rats with overactive bladder (OAB) after cystostomy, and to explore its regulation mechanism on bladder function.

METHODSForty-eight Sprague-Dawley female rats which received cystostomy were randomly divided into a blank group (group A), a blank Sanyinjiao group (group B), a blank non-acupoint group (group C), a model group (group D), a model Sanyinjiao group (group E) and a model non-acupoint group (group F), 8 rats in each one. The model of OAB was established with 1% acetic acid solution perfused into the bladder in the group D, group E and group F. No treatment was given to the group A and group D. Acupuncture was applied at bilateral "Sanyinjiao" (SP 6) in the group B and group E, followed by EA after the arrival of qi. Acupuncture was applied at bilateral non-acupoint in the group C and group F, followed by EA with continuous wave, 2 Hz of frequency for 30 min. The treatment was given for continuous 5 urination cycles. The BL-420 E+ biological function experiment system was used to measure and record the changes of indices of bladder pressure and urodynamics.

RESULTSCompared with the group A, the bladder capacity and urine output in the group B were significantly increased (both P<0.05), and the urination rate was increased in the group C (P<0.05); the differences of each index between group C and group B were not statistically significant (all P>0.05). Compared with the group D, the capacity pressure, bladder capacity, detrusor pressure, urinary output and urination rate in the group E were all increased (all P<0.05). Compared with the group F, the capacity pressure and detrusor pressure in the group E were increased (both P<0.05).

CONCLUSIONThe EA at "Sanyinjiao" (SP 6) could significantly improve urine function in rats with OAB after cystostomy, but its regulation effect on urination is not obvious in rats with non-OAB.

Acupuncture Points ; Animals ; Cystostomy ; Disease Models, Animal ; Electroacupuncture ; Female ; Humans ; Rats ; Rats, Sprague-Dawley ; Urinary Bladder ; physiopathology ; surgery ; Urinary Bladder, Overactive ; physiopathology ; surgery ; therapy

NDV strain ZJ1 strain , a highly virulent NDV strain, has been prevalent among the waterfowls in China mainland in the past years. Multi-basic amino acid sequence distribute in the protease cleavage site of F protein of this strain. Recombinant expressing plasmid pCI-FT, was generated by converting multi-basic amino acid sequence of 112, 115, 117 of the protease cleavage site of F_ 0 protein, to the non-basic amino acid sequence characteristic of avirulent NDV strain. The result from co-expression of mutant or parental F protein with homologous HN protein in COS-1 cells revealed that both mutant and parental F protein had fusion activity. The result from co-expression of mutant or parental F protein with homologous HN protein in CEF cells showed that the cleavage activity of mutant F protein was significantly reduced. The study built a foundation for mutagenesis of amino acid sequence of the protease cleavage site of F_ 0 protein at the full-length cDNA clone level, study on factors contributing to virulence and construction of candidate vaccine strain, and so on.

3T3-LI preadipocytes were induced to differentiate and 3T3-L1 adipocyte or preadipocytes were incubated with oleate or palmitate overnight.RT-PCR was used to measure adiponectin receptor(AdipoR)1 and AdipoR2 mRNA levels.The results showed that the AdipoRl and AdipoR2 expressions were differentiation- dependent.Oleate only suppressed AdipoR mRNA expression in preadipocyte but not in adipocyte.However,high concentration of palmitate reduced AdipoR mRNA expression in both 3T3-LI preadipocyte and adipocyte.

OBJECTIVE: Yang deficiency constitution may bring with different types of illnesses in many people. The present study was designed to observe the effect of ginger-separated snake moxibustion on yang deficiency symptom, and serum adrenocorticotropic hormone (ACTH) and cortisol (CORT) contents in yang deficiency constitution subjects, so as to explore its mechanism underlying improvement of yang deficiency constitution. METHODS: Ninety subjects with yang deficiency type constitution were equally and randomly assigned to control A group (19 men and 26 women, 18－60 years in age) and moxibustion group (16 men and 29 women, 18－60 years in age) and 45 normal subjects with mild constitution (18 men and 27 women, 18－60 years in age) were assigned to control B group. An snake-like moxa-cone was placed at a moxa-holder covering the middle back of human body from Dazhui (GV 14) to Yaoshu (GV 2) after paving a layer (about 1 cm thick) of fresh ginger in the holder, and then ignited, followed by the next moxa-cone, 3 cones altogether every time for each participant. The treatment was conducted once a week, 12 times in total. The subjects' constitution was scored according to the "Classification and Evaluation of Constitution of Traditional Chinese Medicine" formulated by China Association of Chinese Medicine. The serum ACTH and CORT contents were measured by radioimmunoassay before and at the end of the treatment and 6 months after the treatment. RESULTS: The constitation scores of the moxibustion group and control A group were significantly higher than that of the control B group before treatment (P<0.01). At the end of the treatment, the constitution score was significantly decreased in the moxibustion group (P<0.01) rather than in the control A group relevant to its own pre-treatment (P>0.05). The serum CORT and ACTH contents were significantly decreased in both the moxibustion and control A groups in comparison to the control B group before treatment (P<0.01), and obviously increased at the end of treatment in the moxibustion group compared with its own pre-treatment (P<0.01). No significant differences were found among the three groups in the constitution score, and serum CORT and ACTH levels at the 6th month after the treatment compared with their own post-treatment (P>0.05)．. CONCLUSION: The ginger-separated snake moxibustion treatment may improve the constitution of yang deficiency participants by increasing the levels of serum ACTH and CORT, which may be helpful in resisting diseases.

BACKGROUNDRecent studies have demonstrated that dexamethasone (DEX) interferes with immune responses by targeting key functions of dendritic cells (DCs) at the earliest stage. However, the cellular and molecular mechanisms are still incompletely understood. This study aimed to explore the possible mechanisms by investigating the roles of DEX on differentiation, maturation & function of murine DCs and the effects of DEX on DCs via Toll-like receptor 4 (TLR4)-nuclear factor (NF)-kappaB mediated signal pathway.

METHODSImmature DCs (imDCs) were cultured from murine bone marrow (BM) cells. We added DEX into culture medium at different time. The expression of CD11c, CD86 and I-A(b) (mouse MHC class II molecule) was determined by flow cytometry. We determined the expression of NF-kappaB and its inhibitory protein I-kappaBalpha by electrophoretic mobility shift assay (EMSA) and Western blotting, respectively. The productions of interleukin (IL)-12p70 and IL-10 in cell culture supernatants were determined by enzyme-linked immunosorbent assay (ELISA).

RESULTSDEX impaired differentiation of DCs from murine bone marrow progenitors, and inhibited lipopolysaccharide (LPS) induced maturation of DCs. DEX significantly inhibited NF-kappaB expression of normal DCs, the higher the DEX concentration or the longer the DEX treatment time, the more obvious the effect. However, DEX had little effect on LPS-induced NF-kappaB activation, and partially impaired LPS-induced I-kappaBalpha degradation. DEX significantly decreased LPS induced IL-12p70 production by DCs. Interestingly, our results showed a synergistic effect between DEX and LPS on the production of IL-10 by DCs.

CONCLUSIONSDEX inhibits the differentiation and maturation of murine DCs involved in TLR4-I-kappaB-NF-kappaB pathway, and also indirectly impairs Th1 development and interferes with the Th1-Th2 balance through IL-12 and/or IL-10 secretion by DCs.

Animals ; Blotting, Western ; Bone Marrow Cells ; cytology ; Cell Differentiation ; drug effects ; Cells, Cultured ; Dendritic Cells ; cytology ; metabolism ; Dexamethasone ; pharmacology ; Electrophoretic Mobility Shift Assay ; Enzyme-Linked Immunosorbent Assay ; Flow Cytometry ; Male ; Mice ; NF-kappa B ; metabolism ; Signal Transduction ; drug effects ; Toll-Like Receptor 4 ; metabolism

Eight fragments were amplified and cloned into pCR2.1 vector with the designed primers.The fragments,amplified with primer Ⅰ to Ⅶ,were subcloned into transcription vector to construct the plasmid pNDVZJI which contained the full-length cDNA of NDV ZJI strain.The eukaryotic expression vector pCI-L was constructed by subcloning the fragments,amplified with the primer Ⅴ,Ⅵ and Ⅷ,into the expression vector pCI-neo.The full-length cDNA clone,pNDVZJI,with three helper plasmids,pCI-NP、pCI-P and pCI-L,were cotranfected into BSR-T7/5 cell expressing T7 RNA polymerase.After inoculation of transfected cell culture into embryonated chicken eggs from specific pathogen free(SPF)flock,The NDV of ZJI strain was rescued successfully,which laid a good foundation for the further related research.

Based on the complete genome sequence of Newcastle disease virus (NDV) ZJI strain,seven pairs of primers were designed to amplify a cDNA fragment for constructing the plasmid pNDV/ZJI,which contained the full-length cDNA of the NDV ZJI strain.The pNDV/ZJI,with three helper plasmids,pCIneoNP,pCIneoP and pCIneoL,were then cotransfected into BSR-T7/5 cells expressing T7 RNA polymerase.After inoculation of the transfected cell culture supernatant into embryonated chicken eggs from specific-pathogen-free (SPF) flock,an infectious NDV ZJI strain was successfully rescued.Green fluorescent protein (GFP) gene was amplified and inserted into the NDV full-length cDNA to generate a GFP-tagged recombinant plasmid pNDV/ZJIGFP.After cotransfection of the resultant plasmid and the three support plasmids into BSR-T7/5 cells,the recombinant NDV,NDV/ZJIGFP,was rescued.Specific green fluorescence was observed in BSR-T7/5 and chicken embryo fibroblast (CEF) cells 48h post-infection,indicating that the GFP gene was expressed at a relatively high level.NDV/ZJIGFP was inoculated into 10-day-old SPF chickens by oculonasal route.Four days post-infection,strong green fluorescence could be detected in the kidneys and tracheae,indicating that the recombinant GFP-tagged NDV could be a very useful tool for analysis of NDV dissemination and pathogenesis.