[Current status of animal experiment report of acupuncture intervention in chronic fatigue syndrome based on ARRIVE guidelines and GSPC list].

OBJECTIVE: To analyze the current status of animal experiments of acupuncture in intervening chronic fatigue syndrome, so as to search formethods to improve the quality of animal experiment reports. METHODS: From the databases, such as CNKI, VIP, Wanfang, SinoMed, PubMed, Web of Science, Embase and Cochrane Library, the literature of animal experiment on acupuncture treatment for chronic fatigue syndrome was searched from January 1st, 2011 to April 2nd, 2022. Data were extracted according to the animal research reporting in vivo experiment (ARRIVE) guidelines 2.0 and gold standard publication checklist (GSPC), and statistical analysis was performed using Excel 2019. RESULTS: A total of 16 studies were finally included. The satis-faction rate of essential items in the ARRIVE guidelines 2.0 is 41.76%，while the satisfaction rate of recommended items is only 27.73% and of the GSPC is 25.89%. Out of 16 studies, 13 of them explained the reasons for animal exclusion in the experiment, 8 provided specific randomized methods, 8 described detailed information on animal species, strains, and quantities, 3 basically indicated that they had passed ethical review, 7 explained the limitations of the research. All 16 studies reported the main findings and elucidated their potential clinical or scientific value. CONCLUSION: Current animal studies on acupuncture in intervening the chronic fatigue syndrome are of certain limitation. Descriptions of multiple items are incomplete or missing, which prevents rea-ders from assessing reliability and authenticity of the animal experiment. It is recommended that in future research, experimental design, execution and report should be carried out according to the report guidelines for animal experiment to improve research quality.

Mechanism of Key Ingredient of Astragalus membranaceus on Lung Adenocarcinoma via PI3K/AKT Signaling Clarified by Utilizing Network Pharmacology Approach and Experimental Validation.

OBJECTIVE: To investigate the mechanism of the effect of Astragalus membranaceus (A. membranaceus) on lung adenocarcinoma at the molecular level to elucidate the specific targets according to the network pharmacology approach. METHODS: The active components of A. membranaceus and their potential targets were collected from the Traditional Chinese Medicine Systems Pharmacology Database. Lung adenocarcinoma-associated genes were acquired based on GeneCards, Online Mendelian Inheritance in Man (OMIM), PharmGKB, and Therapeutic Targets databases. The PI3K/AKT signaling pathway-related genes were obtained using Reactome portal. Networks of "ingredient-target" and "ingredient-target-pathway-disease" were constructed using the Cytoscape3.6.0 software. The relationships among targets were analyzed according protein-protein interaction (PPI) network. Finally, molecular docking was applied to construct the binding conformation between active ingredients and core targets. Cell counting kit 8 (CCK8) and Western blot assays were performed to determine the mechanism of the key ingredient of A. membranaceus. RESULTS: A total of 20 active components and their 329 targets, and 7,501 lung adenocarcinoma-related genes and 130 PI3K/AKT signaling pathway-related genes were obtained. According to Venn diagram and PPI network analysis, 2 mainly active ingredients, including kaempferol and quercetin, and 6 core targets, including TP53, MAPK1, EGF, AKT1, ERBB2, and EGFR, were identified. The two important active ingredients of A. membranaceus, kaempferol and quercetin, exert the therapeutic effect in lung adenocarcinoma partly by acting on the 6 core targets (TP53, MAPK1, EGF, AKT1, ERBB2, and EGFR) of PI3K/AKT signaling pathway. Expressions of potential targets in lung adenocarcinoma and normal samples were analyzed by using UALCAN portal and found that ERBB2 was overexpressed in lung adenocarcinoma tissues and upregulation of it correlated with clinicopathological characteristics. Finally, quercetin repressed viabilities of lung adenocarcinoma cells by targeting ERBB2 on PI3K/AKT signaling confirmed by CCK8 and Western blot. CONCLUSION: Our finding unraveled that an active ingredient of A. membranaceus, quercetin, significantly inhibited the lung adenocarcinoma cells proliferation by repressing ERBB2 level and inactivating the PI3K/AKT signaling pathway.

[Progress in the application of evaluation criteria for clinical efficacy of acupuncture in the treatment of knee osteoarthritis].

Knee osteoarthritis is a chronic joint disease characterized by degeneration of knee cartilage and secondary osteoproliferation, pain and dysfunction, with a high morbidity.Clinical evaluation of efficacy was mainly based on scales, including pain scales, knee function scales, quality of life scales. In order to fully present comprehensive evaluation criteria of acupuncture effect of knee osteoarthritis, this paper reviewed the scales, contrasted their characteristics and scope of application, analyzed the existing problems, and offered proposals to develop and choose efficacy evaluation criteria. Expecting to provide refe-rence frame to evaluate the clinical efficacy of acupuncture in treating knee osteoarthritis in the future.

[Current state about researches on selection of experimental indexs mechanisms of acupuncture underlying improvement of chronic fatigue syndrome].

Acupuncture therapy is effective in the treatment of chronic fatigue syndrome (CFS) and has its own unique advantages. In the present paper, we reviewed the progress of experimental researches on the underlying mechanisms of acupuncture treatment of CFS in recent 10 years from: 1) regulating the immune system including the peripheral immune organ, immune cells and immune cytokines, proinflammatory and anti-inflammatory cytokines, and lowering the increase of positive rate of multiple mycoplasma infection; 2) regulating the neuroendocrine system including the hypothalamus-pituitary-adrenal axis and stress hormones, monoamine neurotransmitters, and opioid peptides; 3)raising the anti- oxidative stress ability by reducing malondiadehyde, and upregulating activities of antioxidant enzymes superoxide dismutase and glutathione peroxidase; and 4) regulating multiple cellular molecule signaling pathways revealed by genomic and proteomic technologies. In conclusion, acupuncture can relieve CFS through multiple ways and systems, which may provide some ideas for further studies on the biological mechanisms.

[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.

[Research and practice on basic skill training and technique improving of acupuncture and tuina major].

In order to summarize and perfect the basic skill practicing method in acupuncture teaching and form a characteristic teaching, the research on skill training and technique improving of acupuncture and tuina major was carried out. The undergraduate students in acupuncture and tuina major from College of Acupuncture-Moxibustion and Tuina were randomly divided into an experimental group and a control group. Students from both groups received the basic courses of acupuncture-moxibustion and tuina as requested in the syllabus and the special teaching was added to the experimental group, namely "three practice" (strength practice, coordination practice, targeted practice of acupuncture basic skills) and "three enlightenment" (technique enlightenment, mind and qi enlightenment, efficacy enlightenment). During and after the courses, the teaching effectiveness was evaluated on the basis of needling manipulation. The peacetime score in the experimental group was superior to that in the control group (38.03 +/- 1.14 vs 33.25 +/- 1.31, P < 0.05), the total score in the experimental group was superior to that in the control group (84.03 +/- 6.30 vs 78.05 +/- 6.55, P < 0.05). The special teaching method of "three practice" and "three enlightenment" can improve the effect of skill training teaching method. It highlights the unique feature of central plains by integrating the essence of Shaolin internal qigong and Chen-style Taiji, it can also provide new ideas for the practical teaching of TCM professional courses and the construction of characteristic professionals.

[Design and application of hand-wrist caring moxibustion box].

A hand-wrist moxibustion box is designed to effectively solve difficulty to have moxibustion at handwrist joint and problem of bad moxibustion effect as well as improve heat efficiency of moxibustion. The device is consisted of a box and protective screening. The box is hollow with opening on top and bottom. A reversible cover is fixed on the top of the box and support frames are put in the inner-middle. On the side wall there is a hole that is at the same horizontal level of support frames, and horizontal protective screening is put on the bottom. The design of this moxibustion box is novel and unique, simple and reasonable, which could give moxibustion on hand and wrist at the same time. Also it is easily made with low cost, easy to use and favorable effect. It is an innovation on moxibustion box.

[Study on the method of two dimensional polycrylamide gel electrophoresis on rat condylar chondrocyte].

PURPOSE: To investigate the protein profile by two dimensional polycrylamide gel electrophoresis on the rat condylar chondrocyte in vitro. METHODS: The third-passage chondrocytes were harvested from the mandibular condyles of 2-day-old rats in this study. The protein profile of the rat mandibular condylar chondrocytes was examined by two dimensional polycrylamide gel electrophoresis (2-DE-PAGE). The 2-DE gel maps on different pH gradients were obtained. The result of modified coomassi blue-sliver staining and sliver staining was compared using Pdquest 7.1 image analysis software. RESULTS: The results showed that the good protein profile of the condylar chondrocytes was obtained by standard Bio-Rad manual. The protein was mainly in the field from pH4 to pH7. The 1203±86 protein points were examined on 2-DE gel map by modified coomassi blue-sliver staining, and 1769±97 protein points was examined by sliver staining. The silver staining map showed more distinctly but higher background than modified coomassi blue-sliver staining. CONCLUSIONS: The protein profile of the condylar chondrocytes enriches the proteomic database and gives evidence to further proteomic research. The 2-DE map obtained by modified coomassi blue-sliver staining is more suitable for MALDI-TOF mass identification. Supported by National Natural Science Foundation of China (Grant No. C30700963), China Postdoctoral Science Foundation(Grant No.20090461088), Jiangsu Provincial Postdoctoral Science Foundation (Grant No.0802003C) and Nanjing City's Science and Technology Foundation (Grant No.200905011).

Proteomic analysis of early-response to mechanical stress in neonatal rat mandibular condylar chondrocytes.

The objectives of this study were to investigate the early response to mechanical stress in neonatal rat mandibular chondrocytes by proteomic analysis. To evaluate its molecular mechanism, chondrocytes were isolated and cultured in vitro, then loaded mechanical stress by four-point bending system on different patterns. Morphological observation, flow cytometric analysis, and MTT assays indicated that 4,000 microstrain loading for 60 min was an appropriate mechanical stimulus for the following proteome analysis, which produced a transient but obvious inhibitory effect on the cell cycle. Therefore, we took a proteomic approach to identify significantly differential expression proteins in chondrocytes under this mechanical stress. Using 2-DE and MALDI-TOF, we identified seven differentially expressed proteins including the MAPK pathway inhibitor RKIP, cytoskeleton proteins, actin and vimentin, and other selected proteins. Some differentially expressed proteins were validated by both Western blot analysis and fluorescent staining of cytoskeleton at different loading times. The vimentin and RKIP responsive expression were also proven in vivo in oral orthopedic treatment rats, which was in line with the result in vitro. The histological changes in cartilage also showed the inhibition effect. Furthermore, the expressional level of phosphorylated ERK was increased, which demonstrates the changes in MAPK activity. Taken together, these data indicate that mechanical stress resulted in vimentin expression changes first and then led to the subsequent changes in actin expression, MAPK pathway regulated by RKIP and heat shock protein GRP75. All those changes contributed to the cytoskeleton remolding and cell cycle inhibition, finally led to condylar remodeling.

[Effects of acupuncture at "Sishencong" (EX-HN 1) on physiological functions in the sleep disorder model mouse].

OBJECTIVE: To probe into the effect of acupuncture at "Sishencong" (EX-HN 1) on sleep and the mechanism. METHODS: Forty-eight white mice were randomly divided into a Sishencong group, a Zusanli group, a model group and a blank control group, 12 mice in each group. In the mice of the first 3 groups, sleep rhythm disorders was induced by repeatedly regular intraperitoneal injection of small dose of Caffeine, and the mice of the first two groups were treated by acupuncture at "Sishencong" (EX-HN 1) and "Zusanli" (ST 36), respectively. Living state and, circadian activities were observed, and nitric oxide synthase (NOS) activity and nitric oxide(NO) content in the brain were detected. RESULTS: Acupuncture at "Sishencong" (EX-HN 1) or "Zusanli" (ST 36) could significantly increase NOS activity and NO content in the brain, with acupuncture at "Sishencong" (EX-HN 1) being significantly better than acupuncture at "Zusanli" (ST 36) (P < 0.01). CONCLUSION: Acupuncture at Sishencong (EX-HN 1) treats insomnia and regulates sleep and many mental and nervous symptoms, which are carried out partially through increasing NOS activity and NO content, promoting normal function of brain tissues.

[Multiple central clinical studies on the needling method for regulating wei and strengthening brain for treatment of insomnia].

OBJECTIVE: To observe the clinical therapeutic effect of the needling method for regulating wei and strengthening brain on insomnia. METHODS: Two hundred cases of insomnia were randomly divided into a test group and a control group, 100 cases in each group. The test group were treated with the needling method for regulating wei and strengthening brain with Baihui (GV 20), Dazhui (GV 14), Shenmai (BL 62), Zhaohai (KI 6) and ear points Yuanzhong, Shenmen selected; and in the control group, Sishencong (EX-HN 1), Shenmen (HT 7) and Sanyinjiao (SP 6) were selected. Acupuncture was given once daily for 15 days. Pittsburgh sleep quality index (PSQI) was used for scoring before and after treatment. RESULTS: The total effective rate was 89.0% in the test group and 65.0% in the control group with a very significant difference between the two groups (P<0.01); the difference of PSQI scores before and after was -9.15+/-5.68 in the test group and -5.64+/-5.73 in the control group, with a very significant difference before and after treatment in the two groups (P<0.01). CONCLUSION: The therapeutic effect of the needling method for regulating wei and strengthening brain on insomnia is better than that of normal 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.

[Contributions of Yang Ji-zhou to moxibustion methods].

YANG Ji-zhou, a famous scientist of acupuncture and moxibustion in the Ming Dynasty. He was a great master not only in manipulation methods of acupuncture, but also in moxibustion therapy, with unique and profound deep cognition. His main contributions include the three aspects: (1) collected a great deal of methods and experience of moxibustion treatment from moxibustion physicians and gathered and mustered a great works about moxibustion methods before the Ming Dynasty; (2) combined with his own clinical experiences, systematized the methods and techniques of moxibustion, including the relationship between posture and point-fathoming, the treatment orders, the size of moxa cone, the number of moxa cones, the techniques of burning moxa, the methods for vesiculation moxibustion, etc. forming a more systematic manipulation standard; (3) invented and formed YANG's characteristics of clinical moxibustion treatment, i. e selecting a few points to dredge the meridian-qi, grasping opportunity of moxibustion, combining acupuncture and moxibustion with medicine, and consolidating the therapeutic effect by proper diet after treatment.