Inhibitory effect of tanshinone IIA, resveratrol and silibinin on enterovirus 68 production through inhibiting ATM and DNA-PK pathway.

BACKGROUND: Human enterovirus 68 (EV68) is a primary etiological agent for respiratory illnesses, while no effective drug has yet used in clinics largely because the pathogenesis of EV68 is not clear. DNA damage response (DDR) responds to cellular DNA breaks and is also involved in viral replication. Three DDR pathways includes ataxia telangiectasia mutated (ATM), ATM and Rad3-related (ATR), and DNA-dependent protein kinase (DNA-PK). Natural products proved to be an excellent source for the discovery and isolation of novel antivirals. Among them, tanshinone IIA, resveratrol, silibinin, rutin and quercetin are reported to target DDR, therefore their roles in anti-EV68 are investigated in this study. PURPOSE: This study investigated the anti-EV68 ability of various natural compounds related to DDR. STUDY DESIGN AND METHODS: The methods include cell counting, flow cytometry, western blot, Immunofluorescence staining, comet assays, quantitative real-time RT PCR and short interfering RNAs (siRNAs) for analysis of cell number, cell cycle, protein expression, protein location, DNA damage, mRNA level and knock down target gene, respectively. RESULTS: EV68 infection induced DDR. Down-regulation or inhibition of ATM or DNA-PK lowered DDR in EV68-infected cells and mitigated viral protein expression, however, down-regulation or inhibition of ATR unexpectedly up-regulated DDR, and promoted viral protein expression. Meanwhile tanshinone IIA, resveratrol, and silibinin inhibited ATM and/or DNA-PK activation and decreased viral proliferation, while rutin and quercetin inhibited ATR activation and promoted viral production. The role of them in ATM, DNA-PK and ATR activation was consistent with previous reports. CONCLUSION: Tanshinone IIA, resveratrol and silibinin inhibited EV68 proliferation through inhibiting ATM and/or DNA-PK activation, and they were effective anti-EV68 candidates.

[Body acupuncture combined with auricular acupressure for menstrual headache: a randomized controlled clinical trial].

OBJECTIVE: To observe the therapeutic effect of body acupuncture combined with auricular acupressure for treatment of menstrual headache of hyperactivity of "liver fire". METHODS: A total of 85 menstrual headache patients with hyperactivity of "liver-fire" were randomly divided into control group (n = 42) and treatment group (n = 43). Patients of the control group were treated by oral administration of Flunarizine Hydrochloride Capsules (Sibelium, 5 mg/time, twice daily) for 5 days beginning at the headache attack in the first menstrual cycle, and for two weeks (except weekends) in the second and third menstrual cycles. Patients of the treatment group were treated by manual acupuncture stimulation of body acupoints Baihui (GV 20), Fengchi (GB 20), Sanyinjiao (SP 6), Taichong (LR 3), Xiaxi (GB 43), Taiyang (EX-HN 5), Hanyan (GB 4), Xuanlu (GB 5) and Shuaigu (GB 8) beginning from the headache attack, and otopoint-pellet pressure of otopoints unilateral Endocrine, Ovaries, Shenmen, Cortex, Liver, Spleen and Kidney (beginning 5 days before menstrual onset) and bilateral Shenmen, Nie, Liver, Gallbladder, Subcortex and Jiaogan during headache attack. Clinical symptom scores were assessed according to the "Guiding Principles for Clinical Research on New Chinese Herbal Drugs" issued by the Chinese Ministry of Health. RESULTS: After the treatment, of the 42 and 43 cases in the control and treatment groups, 9 (21.43%) and 20 (46.51%) were cured, 12 (28.57%) and 14 (32.56%) had a marked improvement, 13 (30.95%) and 7 (16.28%) were effective, 8 (19.05%) and 2 (4.65%) invalid, with the effective rates being 80.95% and 95.35% , respectively. The curative effect of the treatment group was significantly better than that of the control group (P<0.05). The symptom score and VAS score after the treatment and 3 months post-treatment in the treatment group were significantly lower than those of the control group (P<0.01). The headache scores and VAS scores were significantly reduced following the treatment and 3 months after the treatment in both control and treatment groups (P<0.01). After the treatment for 2-3 therapeutic courses, the headache attack duration during menstruation was markedly and gradually shortened in both groups (P<0.01), and therapeutic effect of the treatment group was significantly better than that of the control group (P<0.01). CONCLUSION: Body acupuncture combined with auricular acupressure treatment can effectively relieve menstrual headache in menstrual women with hyperactivity of "liver fire".

[Effect of electroacupuncture on lipid metabolism in male and female obesity rats].

OBJECTIVE: To investigate the influence of electroacupuncture (EA) on lipid metabolism in both sexes of obesity model rats. METHODS: A total of 30 male and 30 female SD rats were randomized into normal control, model and EA groups respectively, with 10 rats in each group. EA (2 Hz, 1-2 mA) was applied to the ipsilateral "Zusanli" (ST 36)-"Sanyin-jiao"(SP 6) and bilateral "Fenglong" (ST 40) for 30 min, once daily for 28 days. In addition,"Quchi" (LI 11),"Zhongwan" (CV 12) and "Guanyuan" (CV 4) were punctured with filiform needles and stimulated by manipulating the needle for 1 min, separately. Obesity model was duplicated by subcutaneous injection of 15% glutamic sodium (0.2 mL/10 g) once daily for 5 days and by feeding the animal with high fat forage for 36 days. Lee's index was calculated, and plasma triglyeride (TG) concentration detected by glycerol-phosphoric acid oxidase-peroxydase method, total cholesterol (TC) detected by cholesterol oxidase method, and plasma low-density lipoprotein-cholesterol (LDL-C) and high-density lipoprotein-cholesterol (HDL-C) contents were assayed by one-step method, respectively. RESULTS: In both male and female rats, compared to their own normal groups, the Lee's index, plasma TG, TC and LDL-C contents were increased significantly and plasma HDL-C contents decreased obviously in the model groups (P<0.01), while compared to their own model groups, the Lee's index, plasma TG, TC and LDL-C levels in the EA groups were down-regulated considerably and plasma HDL-C contents up-regulated remarkably after the treatment (P<0.01, P<0.05). The efficacies of the EA intervention were obviously better in reducing Lee's index for male rats, and in down-regulating plasma TG and TC contents in female rats (P<0.05). No significant differences were found between male and female rats in EA intervention for plasma LDL-C and HDL-C levels. CONCLUSION: EA intervention can effectively regulate obesity rats' Lee's index and plasma TG, TC, LDL-C and HDL-C levels. The effects are different in male and female rats.

[Therapy of hook needle and manipulations].

With the unique structure and manipulation techniques of blood-letting and cutting, hook needle serves as a role in dredging meridians, removing blood stasis, purging heat for resuscitation and relaxing synechial tissues. There are three needling techniques of hook needle: swift pricking, bleeding and pricking. Six manipulations are commonly used as lifting-thrusting, plucking, pulling, pushing-scraping, vibration and massage. The mechanism and manipulations of hook needling are summarized systematically in the present paper. In this way, its manipulations, indications and contraindications are expounded in detail.

[The history and development of new nine-needle].

This paper introduces the old nine-needle including its name, shape, function, manipulation and origination, as well as systematically introduces the new nine-needle as a new product which has been developed under the guidance of modern science and technology based on the old nine-needle since 1949. As a new tool, the new nine-needle provide much more choice for acupuncture treatment and obtained satisfied clinical effect. During the development of research and clinical practice on the new nine-needle, its theory has been consummated successively. Combining various kinds of needles for treatment is not only paying attention to the specificity of one kind of needles, but also emphasizing the whole function of all kinds of needles, which makes the new nine-needle formed the characteristic and distinct theory of its own.

Sesquiterpene lactones from the root tubers of Lindera aggregata.

Phytochemical investigation of the root tubers of Lindera aggregata resulted in the isolation of five new sesquiterpene lactones, linderagalactones A-E (1-5), along with eight known sesquiterpenoids, 3-eudesmene-1beta,11-diol, hydroxylindestenolide, strychnistenolide, hydroxyisogermafurenolide, atractylenolide III, linderane, neolinderalactone, and linderalactone. The structures and relative configurations of 1-5 were determined by spectroscopic methods, especially HRESIMS and 2D NMR techniques. The absolute configurations of 1-4 were defined by comparison of quantum chemical TDDFT calculated and experimental ECD spectra. Linderagalactone A (1) is a halogenated sesquiterpene lactone possessing a unique rearranged carbon skeleton. Linderagalactone E (5), linderane, hydroxylindestenolide, and linderalactone showed hepatoprotective activity against H2O2-induced oxidative damages on HepG2 cells with EC(50) values of 67.5, 167.0, 42.4, and 98.0 microM, respectively.

[Standardized manipulations of sharp hooked needling for treating migraine and investigation on its mechanisms].

A comparison of recognition to migraine and treatment methods for it was made between traditional Chinese medicine and western medicine in this paper. Emphasis was made on standardized manipulation processes and techniques of sharp hooked needling for treating the disease, based on both the organic conception of the human body, the theory held by TCM, and on the principle of treatment according to pathogenesis obtained through differentiation of symptoms and signs. The necessary preparation before operations, methods for its application and announcements in the standardized manipulations of sharp hooked needling were all explained in detail. Based on its needling, pricking, dissecting and relaxing effects, the mechanisms of sharp hooked needling in treating the disease were investigated. It is concluded that sharp hooked needling is very effective in treating migraine.

[Randomized controlled clinical study on ginger-partitioned moxibustion for patients with cold-damp stagnation type primary dysmenorrhea].

OBJECTIVE: To observe the effect of ginger-partitioned moxibustion in the treatment of patients with primary dysmenorrhea (PD) of cold-damp stagnation type. METHODS: A total of 209 PD patients were randomized into moxibustion group (n=105) and control group (medication group, n=104). Patients of the former group were treated with ginger-partitioned moxibustion at Shenque (CV 8) and Guanyuan (CV 4), once daily for 3 days in the first menstrual cycle, and 3 days before menstruation and once daily for 6 days in the 2nd and 3rd menstrual cycles, and those of control group were asked to take Yueyueshu Granules (a Chinese herbal patent drug for relieving PD). Clinical symptom scores were assessed before and after the treatment. RESULTS: After the treatment, of the 105 and 104 cases in the moxibustion and control groups, the cured, markedly effective, effective and failed cases were 58 (55.24%) and 32 (30.77%), 37 (35.24%) and 33 (31.73%), 5 (4.76%) and 24 (23.08%), and 5 (4.76%) and 15 (14.42%), respectively. The total therapeutic effect of moxibustion group wassignificantly better than that of control group (P<0.01). The effects of moxibustion for relieving mild, moderate and severe dysmenorrhea were evidently superior to those of medication (P<0.01). The symptom scores after the treatment and 3 months of post-treatment in moxibustion group were significantly lower than those of control group (P<0.01). CONCLUSION: Ginger-partitioned moxibustion therapy was effective for PD of cold-damp stagnation type.

[Effects of substance-partitioned moxibustion on plasma beta-EP content in the patient with primary dysmenorrhea of cold-damp stagnation type in the menstrual period].

OBJECTIVE: To probe into the mechanism of substance-partitioned moxibustion in treatment of primary dysmenorrhea (PD) of cold-damp stagnation type. METHODS: The treatment group (105 cases of PD) were treated with substance-partitioned moxibustion and the control group (104 cases) were treated with Chinese drug Yueyue-shu. Their therapeutic effects were observed. Plasma beta-endorphin contents in menstrual period were determined before and after treatment in 40 patients of each group. RESULTS: The total effective rate of 95.2% in the substance partitioned moxibustion group was better than 85.6% in the control group (P < 0.05); after treatment, plasma beta-endorphin content significantly increased in the substance-partitioned moxibustion group (P < 0.01). CONCLUSION: Substance-partitioned moxibustion has obvious therapeutic effect on primary dysmenorrhea of cold-damp stagnation type, which is carried out possibly through regulating the plasma beta-endorphin content as one of the mechanisms.

[Influence of ginger-partitioned moxibustion on serum NO and plasma endothelin-1 contents in patients with primary dysmenorrhea of cold-damp stagnation type].

OBJECTIVE: To explore the mechanism of ginger-partitioned moxibustion in the treatment of cold-damp stagnation type primary dysmenorrhea (PD) patients. METHODS: A total of 209 PD outpatients of cold-damp stagnation type from 3 hospitals were randomized into moxibustion group (n = 105) who were treated with ginger-partitioned moxibustion, and control group (n = 104) who were asked to take Yueyue Shu Granules (an effective patent drug for PD). Ginger-partitioned moxibustion was applied to Shenque (CV 8) and Guanyuan (CV 4) from the menstrual onset on for the first course and 3 days before the onset for the second and third course, continuously for 3 days. Before and after the treatment, plasma endothelin-1 (ET-1) and serum nitric oxide (NO) contents in those patients (n = 40/group) and normal women (n = 20) experiencing menstrual period were determined by radioimmunoassay and nitrate reductase method. RESULTS: After the treatment, of the 105 and 104 cases in moxibustion and control groups, 58 and 32 cured, 37 and 33 markedly effective, 5 and 24 effective, 5 and 15 failed, with the effective rates being 95.24% and 85.58% respectively. The therapeutic effect of moxibustion group was significantly better than that of control group (P<0.05). Before treatment, in comparison with normal group, plasma ET-1 contents in both moxibustion and control groups were significantly higher (P<0.01), while serum NO contents in these two groups markedly lower (P<0.01). After the treatment, self-comparison of both moxibustion and control groups showed that plasma ET-1 decreased significantly and serum NO levels in increased considerably (P<0.05, P<0.01), and the therapeutic effect of moxibustion group was markedly superior than that of control group in lowering plasma ET-1 (P<0.01). CONCLUSION: Ginger-partitioned moxibustion is effective in relieving primary dysmenorrhea patients' abdominal pain, which may be closely associated with its actions in lowering plasma ET-1 level and raising serum NO contents.