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ObjectiveTo explore the possible mechanism of Bimin Formula (鼻敏方) in treating lung-spleen qi deficiency syndrome of allergic rhinitis (AR) with high mucin secretion. MethodsThirty-four SD rats were randomly divided into a blank group (8 rats), a model group (8 rats), a low-dose Bimin Formula group (8 rats), and a high-dose Bimin Formula group (10 rats). Except for the blank group, the other groups were subjected to AR lung-spleen qi deficiency rat models induced by smoking, gavage of Ginkgo biloba leaf extract, and ovalbumin. After modeling, rats in the low- and high-dose Bimin Formula groups were given Bimin Formula concentrate (concentration of 2.16 g/ml) by gavage at doses of 1.08 g/100 g and 2.16 g/100 g, respectively, while rats in the model group were given 0.5 ml/100 g of normal saline by gavage, once daily for 28 days; the blank group was not intervened. Behavioral assessments were performed after intervention. ELISA was used to detect the levels of peripheral blood total immunoglobulin E (IgE). HE staining was used to observe the pathological changes of nasal mucosa epithelium in rats, while immunohistochemistry was used to detect the expression of transmembrane protein 16A (TMEM16A) and mucin 5AC (MUC5AC) protein in nasal mucosa. Western Blot was used to detect the expression of nuclear factor kappa B (NF-κB) protein, and RT-PCR was used to detect the expression of TMEM16A, MUC5AC, and NF-κB mRNA in nasal mucosa. ResultsHE staining showed that the nasal mucosa epithelial cell structure in the blank group was intact without shedding, swelling, or necrosis; the nasal mucosa epithelial tissue of rats in the model group was thickened and partially shed, with infiltration of eosinophils and lymphocytes visible; the pathological changes in nasal mucosa tissue of rats in the high- and low-dose Bimin Formulagroups were improved, and more improvement was showen in the high-dose group. Compared with those in the blank group, the behavioral scores and peripheral blood total IgE levels of rats in the model group significantly increased, as well as the expression of TMEM16A, MUC5AC, and NF-κB proteins and mRNA in nasal mucosa (P<0.05 or P<0.01). Compared with those in the model group, the behavioral scores and peripheral blood total IgE levels of rats in the high-dose Bimin Formula group decreased, and the expression of TMEM16A, MUC5AC, and NF-κB proteins and mRNA in nasal mucosaalso decreased (P<0.05 or P<0.01); the behavioral scores and peripheral blood total IgE levels of rats in the low-dose Bimin Formula group were reduced, and the expression of TMEM16A and MUC5AC proteins and mRNA in nasal mucosa, as well as the expression of NF-κB protein decreased (P<0.05 or P<0.01), but the difference in NF-κB mRNA expression was not statistically significant (P>0.05). Compared with the low-dose Bimin Formula group, the expression of NF-κB protein in the high-dose group decreased (P<0.01). ConclusionBimin Formula may improve the symptoms and high mucus secretion of AR lung-spleen qi deficiency by regulating the TMEM16A/NF-κB/MUC5AC signaling pathway in nasalmucosa.
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OBJECTIVE@#To observe the therapeutic effect of needling technique (acupuncture for regulating spleen and stomach) on diabetic gastroparesis (DGP), and to explore its possible mechanism.@*METHODS@#A total of 128 patients with DGP were randomized into an observation group (64 cases, 4 cases dropped off) and a control group (64 cases, 4 cases dropped off). On the basis of intervention on controlling blood glucose by western medication, needling technique was adopted at Zhongwan (CV 12), Zusanli (ST 36), Yinlingquan (SP 9), Xuehai (SP 10), Sanyinjiao (SP 6), Diji (SP 8), etc. in the observation group, once a day. Mosapride citrate dispersible tablet 5 mg was given orally 3 times a day in the control group. The treatment was given 6 times a week in the both groups, and totally 4-week treatment was required. Before and after treatment, the DGP symptom score, serum content of transmembrane protein 16A (ANO1) were observed, and the clinical therapeutic effect and the safety were evaluated in the both groups.@*RESULTS@#After treatment, the each subitem score (belching, abdominal distension, inappetence, nausea and vomiting, epigastric pain, abnormal defecation) and the total score of DGP symptom were decreased in both groups (<0.05), the subitem scores of belching, abdominal distension, inappetence, nausea and vomiting and the total score in the observation group were lower than those in the control group (<0.05). After treatment, the serum contents of transmembrane protein 16A were reduced in both groups (<0.05), and that in the observation group was lower than the control group (<0.05). The total effective rate was 86.7% (52/60) in the observation group, which was superior to 70.0% (42/60) in the control group (<0.05). Subcutaneous hematoma occurred in 5 cases in the observation group, which was improved after cold compress without other particular intervention.@*CONCLUSION@#The therapeutic effect of needling technique on improving symptoms in patients with diabetic gastroparesis is superior to mosapride citrate dispersible tablet, its mechanism may be related to alleviating the damage of interstitial cells of Cajal (ICC).
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Objective To investigate the expression of transmembrane protein 16A (TMEM16A)in the cochlea of guinea pigs and its relationship with the age-related hearing loss.Methods We used auditory brainstem response (ABR)to explore the changes of hearing in guinea pigs of different age (groups of 2 w,3 m,1 y,and D-galactose).The distribution and expression of TMEM16A in the cochlea were detected by immunofluorescence and Western blot.Results ABR threshold was gradually increased,with significant difference between D-gal and the other three groups (P <0.01).TMEM16A was expressed in the cochlear striae vascularis at different ages,and the expression increased with age before 1 y (P <0.05). However, its level was increased in D-gal group and significantly differed from that in 3 m and 1 y groups (P < 0.05 ).Conclusion The change in TMEM16A expression in the cochlear striae vascularis of guinea pigs may be related to age-related hearing loss.
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Objective To investigate the expression of transmembrane protein 16A (TMEM16A)in the cochlea of guinea pigs and its relationship with the age-related hearing loss.Methods We used auditory brainstem response (ABR)to explore the changes of hearing in guinea pigs of different age (groups of 2 w,3 m,1 y,and D-galactose).The distribution and expression of TMEM16A in the cochlea were detected by immunofluorescence and Western blot.Results ABR threshold was gradually increased,with significant difference between D-gal and the other three groups (P <0.01).TMEM16A was expressed in the cochlear striae vascularis at different ages,and the expression increased with age before 1 y (P <0.05). However, its level was increased in D-gal group and significantly differed from that in 3 m and 1 y groups (P < 0.05 ).Conclusion The change in TMEM16A expression in the cochlear striae vascularis of guinea pigs may be related to age-related hearing loss.
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Calcium-activated chloride channels(CaCCs) play an important role in cardiovascular system,including participating in a variety of physiological functions and being associated with the pathology progress of many cardiovascular diseases.With the development of research,transmembrane protein 16a (TMEM16A) has recently been identified as the molecular basis of CaCCs.Since the study of TMEM16A has got some achievement,especially in the section of cardiovascular system,its main research progress and key points from previous researches are summarized,in which the expression and physiological function of TMEM16A,as well as its clinical pathological correlations with some cardiovascular diseases are told about.Besides,the research prospect of TMEM16A in targeting therapy of cardiovascular disease is also discussed.
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AIM:To investigate the expression of transmembrane protein 16A(TMEM16A) in Fischer rat thy-roid follicular epithelial ( FRT) cells and its electrophysiologic properties .METHODS: The eukaryotic expression vector of pUB6/V5-TMEM16A was constructed and transfected into FRT cells by liposome-mediated transfection .In order to ob-tain the high efficiency of gene transfection and expression , the quantity and ratio of lipid/DNA complexes were optimized . The FRT cells stably expressing TMEM16A were gained by the selection with blasticidin and confirmed by the techniques of RT-PCR and immunofluorescence .The expression and location of TMEM 16A in the FRT cells were observed under an in-verted fluorescence microscope .TMEM16A protein was associated with calcium-dependent chloride current , as measured with halide-sensitive fluorescent protein and patch-clamp technique .RESULTS: The results of double digestion and se-quencing indicated that TMEM16A was cloned into pUB6/V5.The results of RT-PCR and immunofluorescence confirmed that TMEM16A was expressed in the FRT cells after transfection with TMEM16A.The classical calcium-activated chloride channel currents were recorded in the FRT cells stably expressing TMEM 16A by the technique of patch-clamp and halide-sensitive fluorescent protein YFP-H148Q/I152L.CONCLUSION:The protein expression of TMEM16A in the FRT cells was observed.TMEM16A is the molecular identity of calcium-activated chloride channels .