Objective To detect the doubling time of airway smooth muscle cells of human and rat by the xCELLigence in-strument,a real time cellular analyzer.Methods The airway smooth muscle cells were separated by collagenase-pancreatin digestion from the rat airway.Then,added to the different holes of E-plate of xCELLigence instrument with the planting destiny of 3 000 cells/well.and so were the human airway smooth muscle cells.The E-plate was then placed on the xCELLi-gence instrument to monitor cell proliferation for 100 hours to calculate the doubling time by using the RTCA Software Package 2.0 software.Results The doubling time of human airway smooth muscle cell calculated by the real time cellular analyzer was 23.96±0.47 h,which was consistent with the data provided by the reference.The doubling time of rat airway smooth muscle cell was 18.62±0.15 h,and the computational process was simple,time-saving and also effective.Conclusion The xCELLigence instrument can be used to calculate doubling time of airway smooth muscle cells of human and rat, which provides experimental methods and reference data for the basic respiratory disease research.

Objective: To observe the influence on IL-1β and IL-2 in rat models with rheumatoid arthritis after moxibustion on Shenshu (BL 23) and Zusanli (ST 36) points, and to discuss the mechanism of moxibustion. Methods: Fifty male Wistar rats were divided randomly into 5 groups,control group, model group, drug group, moxibustion group, and laser group, 10 for each. Four groups except the normal group were built on the model of rheumatoid arthritis. The changes of body weight and plantar circumference were measured and the level of IL-1β、 IL-2 in sera were examined by ELISA. Results: Compared with the model group, the weight and plantar circumference of rats in the moxibustion group were improved significantly after treatment (P＜0.01), and the improvement of plantar circumference also had significant differences compared with the drug group and the laser group (P＜0.05). The level of IL- 1β、 IL-2 in sera were down regulated in the moxibustion group and the laser group, which had statistical differences compared with the model group (P＜0.05), but no statistical differences were found when comparing with the drug group. Conclusion: Moxibustion obviously improves the toe tumefaction of the rats with rheumatoid arthritis, which is better than CO2 laser of 10.6μm. On the aspect of decreasing the amount of IL-1β、 IL-2, CO2 laser of 10.6 μm is similar with moxibustion.

Objective: To identify the differential proteins in the serum with anti-asthma activity of acupuncture to pinpoint the substance basis of anti-asthma activity of acupuncture by proteomics method. Methods: Two-dimensional electrophoresis of protein was employed to analyze the differential proteins in the serum with anti-asthma activity of acupuncture, and identify the differential proteins with mass spectrum technique. Results: In adrenalectomy (ADX)-asthma acupuncture group and asthma acupuncture group, two-dimensional electrophoresis showed that there was an obvious fuscous point (MW=44.1kD, PI=5.1), which was 60S ribosomal protein L13, or hypothetical protein FLJ32191 analyzed by mass spectrum technique, different from other groups. In ADX-asthma acupuncture group, there were two obviously fuscous points (MW=39.SkD, PI=4.9 and MW=39.SkD, PI=5.0), which were zinc finger protein 208, and thyroid hormone receptor-associated protein analyzed by mass spectrum technique, and two special proteins (MW=17.3kD, PI<4.5 and 16.9kD, PI<4.5), which were Cyclophilin A and zinc finger protein 91 analyzed by mass spectrum technique, different from other groups. Conclusion: There are multi-target effects regulating the whole body in acupuncture treating asthma relating to immunoloregulation, gene expression, and protein synthesization. The effects of acupuncture on the response proteins need further research.

The neuroanatomical basis of meridian effects is one of the focuses of acupuncture mechanism studies. Meridian effects depend mainly on the generation, conduction, integration and output of acupuncture information in the acupoint, spinal cord and brain.The neural tracer technique is a basic method in neurobiology. This article presents recent years' advances in the application of the neural tracer technique to acupuncture field and summarizes the types of neural tracer used in acupuncture studies and regularities in the distribution of neurons and their fibers related to labeled meridians and acupoints in spinal segments and brain regions to provide methodological reference for studies on neurobiological mechanism of acupuncture effects.

Point Dingchuan (Ex-B1) pertains to extraordinary points and often produces a good therapeutic effect on respiratory system diseases. Ancient Chinese medical books do not record point Dingchuan. Modern Chinese medical textbooks describe the location and efficacy of and indications for this point but do not state their provenances. In order to further popularize the clinical application of point Dingchuan, this article verifies the origin and development of the name and location of point Dingchuan so as to provide a certain reference for clinically correct selection and use of this point. The results show that point Dingchuan followed the course of development from ashi point to new point and then to extraordinary point. The development of point Dingchuan to the same acupoint name and location as now international use went mainly through four stages, that is, similar name and different location, similar name and location, same name and different location, and same name and location.

Objective To investigate the effect of acupuncture on the inflammatory changes of α7 nicotinic acetylcholine receptors(α7 nicotinic acetylcholine receptors,α7nAChRs)in rats with chronic migraine(CM).Methods Adult male SD rats were randomly divided into control group(VEH group),model group(NTG group),acupuncture group(TA group),α7nAChRs antagonist group(MLA group)and α7nAChRs agonist group(TA group)according to basic pain threshold.The rat model of CM was established by repeated subcutaneous injection of nitroglycerin(NTG)at the neck and back every other day.TA group and MLA group received acupuncture treatment one hour before NTG injection,20 minutes every day for 9 consecutive days.The MLA group was intraperitoneally injected with α7nAChRs antagonist MLA half an hours before acupuncture,and the PNU group was intraperitoneally injected with α7nAChRs agonist PNU-282987 half an hours before NTG injection,both groups were injected continuously for 9 days.The changes of paw withdrawal mechanical threshold(Paw Withdrawal Mechanical Threshold,PWMT)and tail-flick latency(Tail-Flick Latency,TFL)were detected by Von Frey and thermal radiation pain meter.The contents of inflammatory factors IL-1β,TNF-α and TGF-β in TNC and serum were determined by ELISA.The relative fluorescence intensity and co-expression of GFAP and α7nAChRs in TNC were detected by immunofluorescence dual labeling method.Results Compared with VEH group,PWMT and TFL in NTG group were significantly decreased(P<0.05 or P<0.01),the contents of IL-1β(P<0.01)and TNF-α(P<0.01,P<0.05)in serum and TNC were significantly increased;and in TNC,the astrocytes were activated significantly(P<0.01),while the relative fluorescence intensity of α7nAChRs and the co-expression of GFAP and α7nAChRs were significantly decreased(P<0.05,P<0.01).Compared with NTG group,PWMT and TFL in TA group were increased(P<0.05 or P<0.01),the contents of IL-1β(P<0.01)and TNF-α(P<0.01,P<0.05)in serum and TNC were increased;and in TNC,the relative fluorescence intensity of GFAP was significantly decreased(P<0.01),however the relative fluorescence intensity of α7nAChRs and the co-expression of GFAP and α7nAChRs were significantly increased(P<0.01).Compared with MLA group,PWMT and TFL in TA groups and PNU groups were significantly increased(P<0.05 or P<0.01).The contents of IL-1β(P<0.05)and TNF-α(P<0.01)in serum and TNC of TA group were increased,while the contents of TGF-β(P<0.05)were decreased.The contents of IL-1β in serum and TNC of PNU group were decreased(P<0.01,P<0.05),while the contents of TGF-β(P<0.01)were significantly increased.Conclusions Acupuncture can effectively relieve CM inflammatory response and hypersensitivity to pain,and its anti-inflammatory and analgesic effects may be related to the up-regulation of α7nAChRs expression.