Objective To investigate the expression of anoctamin 1 in Chinese hamster ovary cells (CHO)and to analyze the functional properties of its ion channel,and to provide experimental basis for study on the physiological function of calcium-activated chloride channel.Methods The whole sequence of anoctamin 1 was obtained by PCR technique and was subcloned into pcDNA3.1 to construct the expression vector pcDNA3.1-anoctamin 1 was transfected into CHO by liposome-mediated transfection and the CHO stably expressing anoctamin 1 were aquired by selection with zeocin. The expression and distribution of anoctamin 1 in CHO were measured by RT-PCR technique and inverted fluorescence microscope.The functional properties of anoctamin 1 were measured with halide-sensitive fluorescence proteins YFP-H148Q/I152L.The PBS buffer solution with calcimycin and high concentration of iodine ion was used as experimental group,andthe PBS buffer solution without calcimycin and high concentration of iodine ion was used as control group.Results The results of digestion and sequencing confirmed that anoctamin 1 was cloned into pcDNA3.1 by electrophoresis and blast. The results of RT-PCR and inverted fluorescence microscope indicated that anoctamin 1 was expressed in CHO. The results of I- influx as measured by halide-sensitive fluorescence proteins YFP-H148Q/I152L showed that anoctamin 1 had the more functional properties of trans-epithelial transporting I-,and the transporting speed in experimental group was higher than that in control group (P<0.05).Conclusion Anoctamin 1 can be expressed highly in the CHO;Anoctamin 1 expressed in CHO has the characteristics of calcium-activated chloride channel.

Objective To observe and compare the clinical efficacies between electroacupuncture and warm needling in treating low back pain.Method Seventy-eight eligible low back pain patients were randomized into group A of 28 cases, group B of 26 cases, and group C of 24 cases. Group A was intervened by electroacupuncture, group B was by warm needling, and group C was by medication. The short-form McGill Pain Questionnaire, Japanese Orthopaedic Association Scores (JOA), and Oswestry Disability Index were observed before and after treatment, and the therapeutic efficacies were compared.Result In group A, the McGill item scores [Sensory Pain Rating Index (S-PRI), Affective Pain Rating Index (A-PRI)] respectively after 1-week and 2-week treatment as well as in the 1-month and 3-month follow-up were significantly different from that before treatment (P<0.01,P<0.05). In group B and C, the McGill item scores after 2-week treatment and in the 1-month and 3-month follow-up were significantly different from that before treatment in the same group (P<0.01,P<0.05). The JOA and Oswestry scores were significantly changed respectively after 1-week and 2-week treatment and in the 1-month and 3-month follow-up in the three groups compared with that before treatment (P<0.05,P<0.01). After 1-week and 2-week treatment and in the 1-month and 3-month follow-up, the JOA and Oswestry scores in group A were significantly different from that in group C (P<0.05,P<0.01). In the 1-month and 3-month follow-up, the JOA scores in group B were significantly different from that in group C (P<0.05). The total effective rate was 85.7% in group A and 73.1% in group B, both significantly higher than 58.3% in group C (P<0.05). Conclusion Electroacupuncture and warm needling both can produce a significant efficacy in treating low back pain, but warm needling acts comparatively slowly and is less safe.

OBJECTIVE To predict the quality marker(Q-Marker) of Aurantii Fructus Immaturus in Wendan decoction (WDD) based on fingerprint and network pharmacology. METHODS Establish the fingerprint of Aurantii Fructus Immaturus and WDD reference samples, use Chinese Medicine Chromatographic Fingerprint Similarity Evaluation Software(2012A Edition)for similarity analysis and common component identification; use network pharmacology to screen the related targets and key pathways of common components, and establish the composition-target-pathway network. Predict the potential Q-marker of expectorant stop vomiting, clear gallbladder and stomach of Aurantii Fructus Immaturus in WDD, and determine its content. RESULTS The fingerprints of 15 batches of Aurantii Fructus Immaturus and 15 batches of WDD reference samples were established, and the similarity of fingerprints between 15 batches of Aurantii Fructus Immaturus and WDD reference sample >0.9. Five chemical components were identified, which were naringin, hesperidin, neohesperidin, nobiletin and tangerine. Network pharmacologic screen analysis to getTP53, MAPK8, EGFR, JUN, MMP9 and other 32 core targets of the five compounds and Endocrine resistance, pathways in cancer, and 10 other key pathways. Construct the "component-target-pathway" network diagram. It was predicted that naringin, hesperidin, neohesperidin, nobiletin and tangerine were Q-markers of Aurantii Fructus Immaturus in WDD. The contents of naringin, hesperidin, neohesperidin, nobiletin, tangerine in WDD were not less than 0.011 7%, 0.025 3%, 0.025 7%, 0.010 4%, 0.010 7%. CONCLUSION Q-Marker prediction analysis of Aurantii Fructus Immaturus in WDD provides a reference for establishing a complete WDD quality evaluation system, and also lays a foundation for elucidating the mechanism of its pharmacodynamic material basis.

Migraine is the most common type of primary headache with disabling brain dysfunction. The prevalence of the migraine in Chinese population is 9.3%, and the ratio of female to male is 3∶1, which seriously affect people′s quality of life. Depression is one of the most common psychiatric disorders in migraine comorbidity. Compared with non-migraine patients, the risk of depression comorbidity in migraine patients is more than 2.5 times higher. The frequency and severity of migraine are closely related to depressive symptoms. Depressive symptoms have different effects on headache-related pain signal transduction, which is susceptible to neuroendocrine network disorders in the process of transmission from thalamus to cortex. Neuroendocrine network plays an important role in the depression of migraine comorbidity. Therefore, exploring the pathogenesis of neuroendocrine network in comorbidity provides a theoretical basis for screening more suitable depressive drugs for migraine comorbidity.

Liver is the central hub regulating energy metabolism during feeding-fasting transition. Evidence suggests that fasting and refeeding induce dynamic changes in liver size, but the underlying mechanisms remain unclear. Yes-associated protein (YAP) is a key regulator of organ size. This study aims to explore the role of YAP in fasting- and refeeding-induced changes in liver size. Here, fasting significantly reduced liver size, which was recovered to the normal level after refeeding. Moreover, hepatocyte size was decreased and hepatocyte proliferation was inhibited after fasting. Conversely, refeeding promoted hepatocyte enlargement and proliferation compared to fasted state. Mechanistically, fasting or refeeding regulated the expression of YAP and its downstream targets, as well as the proliferation-related protein cyclin D1 (CCND1). Furthermore, fasting significantly reduced the liver size in AAV-control mice, which was mitigated in AAV Yap (5SA) mice. Yap overexpression also prevented the effect of fasting on hepatocyte size and proliferation. Besides, the recovery of liver size after refeeding was delayed in AAV Yap shRNA mice. Yap knockdown attenuated refeeding-induced hepatocyte enlargement and proliferation. In summary, this study demonstrated that YAP plays an important role in dynamic changes of liver size during fasting-refeeding transition, which provides new evidence for YAP in regulating liver size under energy stress.