MicroRNA-214 protects L6 skeletal myoblasts against hydrogen peroxide-induced apoptosis.

MicroRNAs (miRNAs) have been reported as key gene regulators, and they control many fundamental biological processes. Previously, we demonstrated that miR-214 had a protective effect against myocardial apoptosis and myocardial fibrosis. In this study, we sought to investigate the expression of miR-214 in L6 skeletal myoblast (SKM), the regulatory effect of miR-214 on hydrogen peroxide (H2O2) induced cell apoptosis and the underlying mechanisms of the antiapoptotic effect. MiR-214 expression was up-regulated by H2O2 in a dose and time-dependent manner in L6 SKMs. To investigate the regulatory effects of miR-214 on L6 SKM, both gain-of-function and loss-of-function approaches were applied. The results showed that miR-214 improved cell survival and inhibited cell apoptosis, and blockage of miR-214 abrogated the protective effect on cell survival and resistance to apoptosis. Phosphatase and tensin homolog (PTEN) was negatively regulated by miR-214, and PTEN inhibitor obviously reversed the effect of miR-214 blockage on enhancing cell apoptosis. In addition, miR-214 up-regulated antiapoptotic protein Bcl-2, down-regulated proapoptotic protein Bax, prevented release of cytochrome c and inhibited caspase-3 activation. In summary, H2O2-induced injury increases miR-214 expression in L6 SKM, and miR-214 contributes to the protection of L6 SKM against apoptosis via lowering PTEN and subsequently inhibiting the mitochondrial-mediated caspase-dependent apoptotic signaling pathway.

Transcription factors GATA4 and TBX5 promote cardiomyogenic differentiation of rat bone marrow mesenchymal stromal cells.

Bone marrow mesenchymal stromal cells (BMSCs) have potential applications in cell and gene therapies for cardiac disease. The cardiac-specific transcription factors GATA-binding protein 4 (GATA4) and T-Box protein 5 (TBX5) are considered to be pivotal in cardiogenesis. The aim of this study was to investigate the effects of GATA4 and TBX5 on cardiomyogenic differentiation of BMSCs. The BMSCs were initially isolated and identified. Vectors harboring cardiac transcription factor genes GATA4 and TBX5 or empty vectors were transferred into BMSCs. Cardiomyogenic cells differentiated from BMSCs were identified by expression of cardiac-specific markers including cardiac troponin T, connexin 43, ß-myosin heavy chain, and myosin light chain-2 using immunocytochemical staining, western blotting, and quantitative real-time PCR. The ultrastructures of the differentiated cells were examined by transmission electron microscopy, which were similar to those of fetal cardiomyocytes. The differentiated cells exhibited L-type calcium current activities reflective of the electrophysiological characteristics of cardiomyocytes. These findings indicate that exogenous expression of cardiac-specific transcription factors GATA4 and TBX5 enhance cardiomyogenic differentiation of BMSCs.

[Discussion on professor SHAO Jing-Ming's academic opinion of focusing spirit].

During professor SHAO Jing-ming's academic research and medical practice, his academic opinion of focusing spirit is gradually developed. In terms of nurturing the spirit, attention should be paid on persistence as well as everyday health maintenance and exercise to nurture the spirit of physician. In terms of clinical diagnosis and treatment, patients' psychology, employment and life status should be observed and experienced, which could bring more methods to take essential care of patients' spirit. The treatment should work with psychological counseling, advocating that based on patients' qi and spirit, various forms of treatment methods should be properly used, such as acupuncture or moxibustion or combination of acupuncture and medicine, along with simple acupoint selection and harmony medication. Before clinical treatment of acupuncture, calming the mind is critically emphasized to make a clear diagnosis. During the acupuncture, calming and focusing the mind is necessary as well as emphasizing the details, so acupuncture could be integrated with Chi Gong to create a new warming-sensation technique. In a word, the academic opinion of focusing spirit is shedding an inspiring light upon further study.

[Asthma at acute attack stage treated with "Shao's five needling therapy": a multi-central randomized controlled study].

OBJECTIVE: To evaluate the clinical efficacy of asthma at acute attack stage treated with "Shao's five needling therapy". METHODS: The randomized controlled method was applied to divide 210 cases into an observation group and a control group, 105 cases in each one. In the observation group, "Shao's five needling therapy" [Feishu (BL 13), Dazhui (GV 14), Fengmen (BL 12)] and the combined therapy were adopted, including oxygen uptake, aerosol inhalation and oral administration of prednisone. In the control group, the oral administration of theophylline sustained release tablet and the combined therapy were applied. The treatment was continued for 7 days. The clinical symptoms and physical signs such as wheezing, cough, expectoration, chest stuffiness, wheezing rale and shortness of breath, as well as lung function indices such as forced expiratory volume one second (FEV1) and peak expiratory flow (PEF) were observed before and after treatment in the two groups. METHODS: In the observation group, 69 cases were cured clinically, 20 cases effective remarkably, 7 cases effective and 0 case failed. In the control group, 49 cases were cured clinically, 31 cases effective remarkably, 15 cases effective and 0 case failed. The difference in the efficacy was significant in comparison of the two groups (P < 0.05). The therapeutic effect in the observation group was better than that in the control group. The total score of the symptoms and physical signs, FEV1 and PEF after treatment were all improved significantly in the two groups (all P < 0.01). And the results in the observation group were better than those in the control group (all P < 0.01). CONCLUSION: "Shao's five needling therapy" achieves the significant efficacy on asthma at acute attack stage. It significantly relieves the symptoms and physical signs of the patients and improves lung functions. The effect is better than that of theophylline sustained release tablet.

Diazoxide pretreatment enhances L6 skeletal myoblast survival and inhibits apoptosis induced by hydrogen peroxide.

Skeletal myoblast (SKM) transplantation is a promising approach to regenerate tissue and improve the function of the injured heart. However, the number of survival cells transplanted to host myocardium is quite poor due to high rate of apoptosis; diazoxide (DZ) is a highly selective mito-KATP channel opener that may reduce cell apoptosis by relieving reactive oxygen species (ROS) damage. The aim of this study is to explore the protective effects of DZ on L6 SKM damage induced by hydrogen peroxide (H(2)O(2) ) in vitro. Different dose and time of H(2)O(2) and DZ treatment were performed and only 24 hr of 1.00 mmol/L H(2) O(2) treatment and 200 µmol/L DZ pretreatment for 30 min were used for further experiment. L6 SKMs were cultured and divided into control group (no treatment), H(2)O(2) group (24 hr of 1.00 mmol/L H(2) O(2) treatment) and DZ + H(2)O(2) group (pretreated with 200 µmol/L DZ for 30 min before 24 hr of 1.00 mmol/L H(2) O(2) treatment). Compared with control group, H(2)O(2) treatment caused cell damage, increased lactate dehydrogenase release, cell apoptosis, and bax gene expression, while reduced cell proliferation and decreased bcl-2 expression. DZ pretreatment may protect cells from damage induced by H(2)O(2) and reduce cell apoptosis by increasing bcl-2 and decreasing bax expression. DZ pretreatment may also promote cell proliferation measured by both PCNA expression and flow cytometry method. These results suggest that DZ may protect L6 SKMs from damage induced by H(2)O(2) by maintaining integrity of cell membrane, reducing apoptosis and increasing proliferation in vitro.

[Professor Shao Jing-ming: a famous expert of acupuncture and moxibustion].

Professor SHAO Jing-Ming devotes his life in clinical practices for more than 80 years. He is an eminent specialist acupuncture with abundant clinical experiences. He establishes the unique method of needling of tri-points and penta-needles which achieves significant effectiveness for the treatment of asthma. He has taught for more than 50 years with unique educational method. A great amount of experts appear in the circle of TCM after receiving his unique training. He devotes his life to composing books on acupuncture and moxibustion. He devotes his life and his heart to promoting the development of acupuncture and moxibustion, and makes remarkable contribution in this field.

[Multi-central clinical valuation of SHAO's "five needling methods" for treatment of asthma of deficiency of lung and spleen].

OBJECTIVE: To make multi-central clinical valuation of SHAO's "five needling methods" for treatment of asthma of deficiency of lung and spleen. METHODS: Two hundred and ten cases were randomly divided into a test group and a control group, 105 cases in each group. The test group were treated with SHAO's "five needling methods", with Feishu (BL 13), Dazhui (GV 14), Fengmen (BL 12) selected; the control group were treated with routine needling method, with Dingchuan (EX-B 1), Gaohuang (BL 43), Feishu (BL 13), Taiyuan (LU 9), Pishu (BL 20), etc. selected. The treatment in the two groups was given once each day, for 4 weeks, with one-day interval each 6-consecutive day. RESULTS: Fourteen cases were clinically cured, 42 cases were markedly effective, 32 cases were effective and 6 cases were ineffective in the test group with an effective rate of 93.6%; and 8, 30, 41 and 13 cases in the control group, respectively, with an effective rate of 85.9%, with a significant difference between the two groups (P < 0.05). After treatment, the symptoms and signs, pulmonary function in the test group were better than those in the control group (P < 0.05). CONCLUSION: SHAO's "five needling methods" has significant therapeutic effect for treatment of asthma of deficiency of lung and spleen, which is better than that of the routine needling method.

[Study on professor Shao Jing-ming's academic thought].

Professor Shao Jing-ming has been engaged in medicine for over 70 years. He has original academic view in diagnosis of diseases, prescription of combined acupoints, needling manipulation and combination of acupuncture and moxibustion with medication, etc.. He holds that disease should be combined with syndrome, modern medical sciences are used for definite diagnosis of diseases; for acupoint prescriptions, fewer and better acupoints are selected, and treatment of difficult and complicated diseases with back-shu points usually can attain good results; correct application of needling manipulation and Deqi is the key of therapeutic effects; he creates and uses heat-sensation needling manipulation, the method of twirling-inserting needle with finger pressure, injection-type inserting needle method; combination of acupuncture-moxibustion with medicine can increase therapeutic effects. He writes a book Zhongyi Zhiyao of about 300,000 words.

[Study on the law of professor SHAO Jing-ming's clinical point selection].

Professor Shao Jing-ming forms an unique academic thought and technique specialty on the basis of carefully study on medical classics, widely extensive learning, and diligent practice. In several ten years' clinical work, he uses TCM theories as basis and channel theories as guidance, following the standards of treatment based on syndrome differentiation, and in clinical acupuncture and moxibustion treatment, he emphasizes "selection of primary and secondary acupoints, and needling in order of priority". In clinical treatment of various diseases, 3 points are commonly used as main points, with accurate and proper selection of acupoints.