Effect of the activation of P2X7 receptor on D-serine release in ventrolateral periaqueductal gray in rats with bone cancer pain / 医学研究生学报

Objective　The activation of P2X7 receptor in ventrolateral periaqueductal gray (vlPAG) is involved in the formation and maintenance of bone cancer pain (BCP). This study will establish a rat model of BCP and observe the effect of the activation of P2X7 receptor in vlPAG on D-serine level through brain microdialysis combined with ELISA.　Methods　Forty-two female SD rats were divided into four groups by random number table: normal control group (n=12), sham group (n=12), BCP group (n=12) and P2X7 receptor antagonist group (n= 6). The model of metastatic BCP in the tibias of the rats was established in the BCP group, and 20μL of RPMI-1640 medium cell suspension containing SHZ-88 breast cancer cells was injected (1×107 cancer cells/0.5 mL). The sham group was injected with treated cancer cells of the same volume (SHZ-88 breast cancer cells were kept in boiling water at 90 ℃ for 20 min), and the rest of the operation was the same as the BCP group. The normal control group received no treatment. The P2X7 receptor antagonist group was treated the same as the BCP group, except that the P2X7 receptor-specific antagonist A-438079 was added to the perfusion solution. The thermal pain threshold and mechanical pain threshold were detected at the same time in the normal control group, the sham group and the BCP group. The positive expression of P2X7 receptor in vlPAG of rats was detected by immunohistochemistry in each group in 21 days. The changes of D-serine in vlPAG dialysate were detected by ELISA in each group.　Results　The mechanical pain threshold and thermal pain threshold of the rats in BCP group on Day 5, 7, 10, 14, 18 and 21 were lower than those of the normal control group and sham group (P<0.01). The positive expression of P2X7 was scattered in vlPAG in normal control group and sham group. The number of P2X7 receptor positive cells in the BCP group was significantly higher than that in the control group and sham group (P<0.01). The content of D-serine in vlPAG of the rats in BCP group [(220.28±63.38)ng/mL] was significantly higher than that in the control group [(148.09±46.89)ng/mL] and the sham group [(147.32±51.44)ng/mL] (P<0.05). The content of D-serine in vlPAG [(134.20±41.77)ng/mL] in P2X7 receptor antagonist group was significantly lower than that in BCP group (P<0.05).　Conclusion　The activation of the P2X7 receptor in ventrolateral periaqueductal gray promotes D-serine release and participates in the mechanisms of BCP in rats .

The effect of activation of P2X7 receptor on Ca2+ level in lateral midbrain periaqueductal gray neuron / 医学研究生学报

Objective The main intracellular signal of P2X7 receptor activation is the increasing of Ca2+, which then presents the diversity of its physiological and pathological functions through multiple intracellular signal transduction. To observe the effect of activation of P2X7 receptor on intracellular calcium（Ca2+）level in lateral midbrain periaqueductal gray (lPAG) neurons of primary cultured rat. Methods The primary cultured lPAG neurons were randomly divided into 4 groups: control group(no drug added), only for control; BzATP group(100 μmol/L); A-740003+ BzATP group(incubate with 100 nmol/L A-740003 for 10 min, then add 10 μmol/L ofBzATP); BzATP control group(add in Ca2+-free solution for 20 min, then add BzATP). The incubation solution of control group, BzATP group and A-740003+ BzATP group are DMEM/F12 medium, and the BzATP control group is Ca2+-free. The laser scanning confocal microscopy (LSCM) was used to detect : the changes of cultured neuron Ca2+ levels by different concentrations of BzATP; the effects of A-740003 and Ca2+-free medium preincubation on BzATP-induced Ca2+ level alterations in cultured neurons. Results BzATP dose-dependently increased the Ca2+ levels in cultured lPAG neurons; A-740003 and Ca2+-free medium inhibited the BzATP-induced increasing of Ca2+ level in cultured lPAG neurons. LSCM showed: The intracellular calcium fluorescence insensity(2.48±1.05) in the BzATP group was significantly higher than that in the blank control group, BzATP control group and A-740003+ BzATP group[(1.12±0.03), (1.09±0.03), (1.14±0.08)](P<0.01). Conclusion The activation of P2X7 receptor can increase the level of lPAG neurons Ca2+ , and is associated with the extracellular Ca2+ influx.

HPLC determination of the content and dissolution of isophenylcyclopentylamine hydrochloride capsules / 国际药学研究杂志

Objective To establish a method for the determination of the content and dissolution of isophenylcyclopentyl?amine hydrochloride capsules. Methods The HPLC analysis was performed on a Diamonsil C18 column(150 mm×4.6 mm,5μm). A mixture of 0.02 mol/L KH2PO4 solution containing 0.1%triethylamine with pH adjusted to 3.0 by phosphoric acid-methanol-acetonitrile (30:35:35)was used as the mobile phase with the flow rate at 1.0 ml/min. The detection wavelength was 224 nm. Dissolution was de?termined by the basket method,using the 500 ml of 0.1 mol/L hydrochloric acid solution,pH 4.5 acetic acid buffer,water and pH 6.8 phosphoric acid buffer as dissolution media under the 50,75 and 100 r/min rotation speeds to select the dissolution condition. Re?sults This method had high specificity. The linear range for the quantitative determination was 20.74-155.58 μg/ml(r=1.0000), and the average recovery was 100.1%. The linear range for the determination of dissolution was 2.08-24.90μg/ml(r=0.9998),with the average recovery of 98.9%. The method of dissolution tests was established:0.1 mol/L hydrochloric acid solution was used as disso?lution medium and rotation speed was 50 r/min. The determined content and dissolution of three batches of capsules fulfilled the re?quirements. Conclusion The method is simple,accurate and reproducible for the determination of the content and dissolution of iso?phenylcyclopentylamine hydrochloride capsules.

Electroacupuncture Ameliorates Cerebral Ischemia-Reperfusion Injury by Regulation of Autophagy and Apoptosis.

Background. The therapeutic mechanisms of cerebral ischemia treatment by acupuncture are yet not well addressed. Objective. We investigated the effects of electroacupuncture (EA) at GV26 observing the expression of autophagy-related proteins Beclin-1 and LC3B and proportion of apoptotic cells and Bcl-2 positive cells in MCAO/R model rats. Methods. Sprague-Dawley (SD) male rats were randomly assigned to 7 groups: model groups (M6h, M24h, and M72h), EA treatment groups (T6h, T24h, and T72h), and sham operation group (S). Neurological deficit and cerebral infarction volume were measured to assess the improvement effect, while the expression of Beclin-1 and LC3B and proportion of Tunel-positive and Bcl-2 positive cells were examined to explore EA effect on autophagy and apoptosis. Results. EA significantly decreased neurological deficit scores and the volume of cerebral infarction. Beclin-1 was significantly decreased in T24h, while LC3B-II/LC3B-I ratio markedly reduced in 6th hour. EA groups markedly reduced the number of Tunel positive cells, especially in T24h. Meanwhile, the number of Bcl-2 positive cells obviously increased after EA treatment, especially in T6h and T24h. Conclusions. The alleviation of inadequate autophagy and apoptosis may be a key mechanism involved in the reflex regulation of EA at GV26 to treat cerebral ischemia.

Wrist-ankle acupuncture (WAA) for precompetition nervous syndrome: study protocol for a randomized controlled trial.

BACKGROUND: Precompetition nervous syndrome comprises an excessive nervous and anxiety response to the high-pressure environment preceding a sporting competition. The use of acupuncture as a treatment option for anxiety, and wrist-ankle acupuncture (WAA) specifically in this instance, has been identified as a growing trend within the Western world. In our previous study, we have confirmed the efficacy of WAA for pre-examination anxiety. In this paper, we present a randomized controlled single-blind trial evaluating the use of WAA for precompetition nervous syndrome, comparing it with the intervention of sham acupuncture. METHODS/DESIGN: The study was designed as a randomized controlled single-blind trial to evaluate the effects of WAA for precompetition anxiety. The trial will be conducted in annual track and field events of Shanghai University of Sport. A total of 100 participants who meet inclusion criteria are randomly assigned by computerized randomization to receive WAA therapy or sham acupuncture. The group allocations and interventions are concealed to participants and statisticians. The Competition State Anxiety Scale (CSAI-2) is used as the primary outcome measure, while heart rate, blood pressure, respiratory frequency, tension syndrome curative effect evaluation and participants' feeling of acupuncture questionnaire are applied as secondary outcome measures. DISCUSSION: The results of this trial will confirm whether WAA is effective to treat precompetition anxiety in annual track and field events. TRIAL REGISTRATION: Chinese Clinical Trial Registry (identifier: ChiCTR-TRC-13003931; registration date: 22 October 2013).

[Efficacy of ginsenosides combined with prednisone in patients with systemic lupus erythematosus: a prospective, randomized, double-blind, placebo-controlled trial].

BACKGROUND: The side effects of glucocorticoid in treatment of systemic lupus erythematosus (SLE) have been the focus of debate, and our preliminary study indicates that ginsenosides can enhance the efficacy of dexamethasone. OBJECTIVE: To observe the effects of ginsenosides combined with prednisone in SLE patients. DESIGN, SETTING, PARTICIPANTS AND INTERVENTIONS: A total of 60 SLE patients from Department of Rheumatology and Immunology, Changhai Hospital, Second Military Medical University, were randomly divided into treatment group and control group, with 30 patients in each group. Patients in the treatment group were given routine treatment with prednisone plus ginsenosides, while those in the control group were given routine treatment with prednisone plus placebo. They were all treated for 3 months. MAIN OUTCOME MEASURES: After three-month treatment, syndrome score in traditional Chinese medicine (TCM), total response rate and symptom improvement rate were measured and evaluated. RESULTS: Twenty-eight cases in treatment group and twenty-seven cases in control group were included in analysis. The total response rates in the treatment group and control group were 89.28% and 66.67% respectively, and there was a significant difference between the two groups (P<0.05). After treatment, the TCM syndrome scores in the two groups were lower than those before treatment (P<0.01), and prednisone plus ginsenosides was better in decreasing the TCM syndrome score than prednisone plus placebo (P<0.05). The symptoms were improved in the treatment group as compared with the control group (P<0.05). CONCLUSION: Prednisone combined with ginsenosides can increase the clinical effective rate and improve the clinical symptoms of SLE patients.

[Efficacy of combined therapy with ginsenosides and prednisone in treating systemic lupus erythematosus--a randomized, controlled and double-blinded trial].

OBJECTIVE: To observe the therapeutic efficacy of combined therapy with ginsenosides (GS) and prednisone on systemic lupus erythematosus (SLE). METHODS: Sixty patients with SLE were assigned to 2 groups randomly by a randomizing digital table, the treated group and the control group, 30 cases in each group. All patients were treated with routine administration of prednisone, but to the treated group GS Capsule (50 mg) was given additionally twice every day, while to the control group placebo capsule of equal dosage was given instead. The total clinical efficacy, and changes of systemic lupus erythematosus disease activity index (SLEDAI), erythrocyte sedimentation rate (ESR), complement 3 (C3) and anti-ds-DNA were observed after 3-month-treatment. RESULTS: The total clinical efficacy rate was 89.28% in the treated group and 66.67% in the control group, showing a significant difference between them (P<0.05). The improvements of SLEDAI, ESR and C3 in the treated group were more significant than those in the control group (P<0.01, P<0.05). CONCLUSION: The combined therapy of GS and prednisone could enhance the clinical efficacy, reduce the SLEDAI and promote the recovery of laboratory indices in SLE patients.