SKF96365 impedes spinal glutamatergic transmission-mediated neuropathic allodynia

Spinal nerve injury causes mechanical allodynia and structural imbalance of neurotransmission, which were typically associated with calcium overload. Storeoperated calcium entry (SOCE) is considered crucial elements-mediating intracellular calcium homeostasis, ion channel activity, and synaptic plasticity. However, the underlying mechanism of SOCE in mediating neuronal transmitter release and synaptic transmission remains ambiguous in neuropathic pain. Neuropathic rats were operated by spinal nerve ligations. Neurotransmissions were assessed by whole-cell recording in substantia gelatinosa. Immunofluorescence staining of STIM1 with neuronal and glial biomarkers in the spinal dorsal horn. The endoplasmic reticulum stress level was estimated from qRT-PCR. Intrathecal injection of SOCE antagonist SKF96365 dose-dependently alleviated mechanical allodynia in ipsilateral hind paws of neuropathic rats with ED 50 of 18 μg. Immunofluorescence staining demonstrated that STIM1 was specifically and significantly expressed in neurons but not astrocytes and microglia in the spinal dorsal horn. Bath application of SKF96365 inhibited enhanced miniature excitatory postsynaptic currents in a dosage-dependent manner without affecting miniature inhibitory postsynaptic currents. Mal-adaption of SOCE was commonly related to endoplasmic reticulum (ER) stress in the central nervous system. SKF96365 markedly suppressed ER stress levels by alleviating mRNA expression of C/ EBP homologous protein and heat shock protein 70 in neuropathic rats. Our findings suggested that nerve injury might promote SOCE-mediated calcium levels, resulting in long-term imbalance of spinal synaptic transmission and behavioral sensitization, SKF96365 produces antinociception by alleviating glutamatergic transmission and ER stress. This work demonstrated the involvement of SOCE in neuropathic pain, implying that SOCE might be a potential target for pain management.

Establishment of HPLC Fingerprints and Content Determination of 7 Components of Jingu Tongxiao Pill / 中国药房

OBJECTIVE：To establish the fingerprint of Jingu tongxiao pill ，and to determine the contents of 7 components. METHODS：HPLC method was adopted. The determination was performed on Cosmosil 5C18-MS-Ⅱ column with acetonitrile- 0.02％ phosphoric acid solution as mobile phase （gradient elution ）at the flow rate of 1.0 mL/min. The detection wavelength was 230 nm and the column temperature was set at 25 ℃. The sample size was 10 μL. HPLC fingerprint of 10 batches of Jingu tongxiao pill was established by using Similarity Evaluation System of TCM Chromatographic Fingerprint （2012 edition），and common peak was identified by comparing with the mixed reference substance. The contents of corresponding components of the identified common peak were determined by the same HPLC method. RESULTS ：There were 20 common peaks in HPLC fingerprint of 10 batches of samples ，and the similarity with the control fingerprint was not less than 0.980. By comparing with the mixed reference substance， 7 components were identified ， which were loganic acid ，gentiopicroside，paeoniflorin，salvianolic acid B ， cryptotanshinone，tanshinone Ⅰ and tanshinone Ⅱ A. The linear range of the above 7 components were 4.509-45.090， 15.090-150.900，14.985-149.850，14.982-149.820，2.967-29.670，1.944-19.440，3.094-30.940 μg/mL（all r＞0.999），respectively. The limits of detection were 0.060 1，0.161 0，0.399 6，0.159 8，0.031 6，0.051 8，0.082 5 μg/mL，respectively. The limits of quantitation were 0.200 4，0.503 0，0.999 0，0.399 5，0.079 1，0.259 2，0.412 6 μg/mL，respectively. RSDs of precision ，stability （24 h） and reproducibility tests were all lower than 3.0％ （n＝6）. Average recoveries were 98.81％ -100.28％ ，RSDs were 0.20％-1.21％（n＝6）. In 10 batches of samples ，the contents of the above 7 components were 0.441 0-0.969 4，3.283 4-4.733 4， 1.947 7-3.674 9，1.336 6-2.270 9，0.293 2-0.372 1，0.190 2-0.293 9 and 0.352 8-0.518 8 mg/g，respectively. CONCLUSIONS ：In this study，HPLC fingerprint and content determination method of Jingu tongxiao pill are successfully established and can be used for quality control.

Preparation and Quality Control of Aspirin-β-cyclodextrin-PLGA Microspheres / 中国药房

OBJECTIVE:To prepare aspirin-β-cyclodextrin-PLGA microspheres,and control its quality. METHODS:Aspirin-β-cy-clodextrin inclusion complexes were firstly prepared,and then aspirin-β-cyclodextrin-PLGA microspheres were prepared by emul-sion-solvent evaporation method. The morphology and particle size of microspheres were detected,and entrapment efficiency and accu-mulative release rate were calculated. With entrapment efficiency as index,orthogonal test was adopted to optimize stirring speed,PVA concentration,PVA volume and feed ratio. RESULTS:The optimal formulation was as follows as stirring speed of 4 000 r/min,PVA concentration of 3%(g/100 ml),PVA volume of 30 ml,feed ratio of 1∶10. Prepared microspheres were round and smooth in appear-ance. Entrapment efficiency of the microspheres was (41.79 ± 1.09)%. The diameter were regular and ranged 0.5-127.5 μm. As drug-loaded microspheres degraded,the release of aspirin was slow and its accumulative release rate was 83%within 600 h. CONCLU-SIONS:Aspirin-β-cyclodextrin-PLGA microspheres are prepared successfully with regular morphology and good sustained-release.

Clinical Study of Low Molecular Weight Heparin Combined with Alteplase for Thrombolysis / 昆明医科大学学报

Objective The aim of this study was to investigate the application of low molecular weight heparin combined with alteplase for thrombolysis in patients with acute myocardial infarction(AMI),and to compare its thrombolysis effect with traditional anticoagulant-general heparin. Methods 35 AMI patients with alteplase intravenous thrombolytic therapy were divided into two groups,among which 15 cases in the study group were treated with 30 mg enoxaparin instant intravenous injection before thrombolysis,40 mg enoxaparin hypodermic injection 15 minutes after thrombolysis,40 mg enoxaparin hypodermic injection every 12 hours for 5 days. The other 20 cases were assigned to the control group and treated with 4 000 u(or 60 u/kg)general heparin instant intravenous injection,then pumped general heparin intravenously in the rate of 800-1 000 u/h. Meanwhile,detected coagulation convention every 3 hours and adjusted the dose of heparin according to the activated partial thromboplastin time(APTT)to keep APTT value within 1.5 to 2.0 times(50-70 s)the standard value. 48 hours later,patients in the control group was given hypodermic injection of 40 mg enoxaparin every 12 hours for 5 days. All cases were given regular treatment as guidelines recommended. Results The patency rate of infarct-related coronary artery assessed by unified clinical criteria was 86.66% and 65.00%,and the average recanalization time was(1.62±0.36)h and(1.81±0.33)h in the study and control groups,respectively. There were no significant differences on patency rate and recanalization time between the two groups(P > 0.05). One case of bleeding complication occurred in the study group(6.66%),while five cases occurred in the control group(25.00%). In the two groups,no cases appeared intracranial hemorrhage and other serious or fatal bleeding occurred. Conclusion The clinical efficacy of low molecular weight heparin was non-inferior to general heparin in AMI patients with alteplas thrombolysis. The application of low molecular weight heparin was simple and easy to operate. It can optimize the procedure of thrombolysis,reduce repetitive coagulation index monitoring and lessen clinical workload,which is worth being promoted to primary hospital and the emergency systems.

Study on Quality Standard for Huamoyan Tablet / 中药新药与临床药理

Objective To establish the quality standard for Huamoyan Tablet. Methods Radix Salviae MiltioiThizae and Radix Cyathulaewere were identified by TLC. The content of protocatechuic aldehyde was determined by HPLC. Results The spots on TLC plates were clear without the interference of negative control. Protocatechuic aldehyde showed a good linear relationship in the range of 0. 021～0. 105 μg, r=0. 999 9. The average recovery was 98. 13 % ,and RSD=2. 41%. Conclusion The method is simple, sensitive and accurate, and can be used to control the qual-ity of Huamoyan Tablet.

Pharmacodynamical research of Jingu Tongxiao granule / 中国中药杂志

<p><b>OBJECTIVE</b>To study functions of Jingu Tongxiao granule (JGTXG, treatmenting ache of bones and muscles) in antiphlogistic and antalgic aspect, invigorating the circulation of blood and absorbing clots and antitraumatic soft tissue.</p><p><b>METHOD</b>Animal models of inflammation, ache, gore and traumatic soft tissue were adopted, and pharmacodynamic actions of Jingu Tongxiao granule were observed.</p><p><b>RESULT</b>JGTXG could conspicuously restrain inflammatory reactions of mouse ear tumid model treated by croton oil tumid and rat foot metatarsus tumid model treated by carrageenan, and restrain pain responses of mouse caused with whipping back end method by heat stimulating and of mouse caused with wriggling body method by acetic acid being injected in its abdominal cavity. It could significantly improve petechia degree in traumatic rat blood stasis model, and prominently improve raumatized limb's tumefaction degree and alleviate blood stasis, swelling and phlogistic cell soakage in traumatic rat soft tissue model. At the same time, it could prominently restrain platelet aggregation and improve whole blood viscosity.</p><p><b>CONCLUSION</b>Jingu Tongxiao granule has antiphlogistic and antalgic functions, invigorating the circulation of blood and absorbing clots and antitraumatic soft tissue, and it could keep curative effect of original dosage form.</p>

Studies on Quality Standard of Wanbile Wan / 中药新药与临床药理

Objective To set up the quality standard of Wanbile Wan. Method s Fructus Psoraleae, Radix Puerariae, Radix paeoniae Alba and Radix Glycyrrhiza e Preparatae in Wanbile Wan were identified by TLC .Isopsoralen was determined b y TLC scanning. The chromatographic conditions were: high performance silica gel G plate as lamellated plate, n- hexane - ethyl acetate (8 ∶ 2)being the exp ander, serrated scanning with single- wavelength reflection, beam stricture of 0.4? 0.4 mm, linear parameter SX=3 and scanning wavelength at 300 nm. Results The TLC spots were highly specific, clear and concentrated without interferen ce of the presence of negative controls. Isopsoralen showed a good linearity wit hin the range of 0.064～ 1.536 ? g, r=0.9980, average recovery rate was 100.7 % , and RSD was 2.0% .Conclusion This method can be used for the quality co ntrol of Wanbile Wan.

GC/MS fingerprint of Jingu Tongxiao Pills / 中成药

AIM:To establish a GC/MS method for determining the fingerprint of Jingu Tongxiao Pills.METHODS:GC/MS condition was comprised of column:DB-1(0.25 mm?30 m,0.25 ?m);injector temperature of 250 ?C;interface temperature of 250 ?C;column temperature of 70?C(1min)4?C/min→130?C(15min)1?C/min→150?C10?C/min→250?C(7min);carrier gas:He;column pressure:60 kPa;ionization souse:EI;detector volts:1.00 kV;mass range:41～350 aum.RESULTS:17 peaks existed on the GC/MS fingerprint of Jingu Tongxiao Pills.CONCLUSION:The method can provide more information for the quality control of Jingu Tongxiao Pills.

Effect of Dangui Huoxue Zhitong Granule on Analgesia,Anti-inflammation and Anti-injury of Soft Tissues / 中医杂志

Objective To observe the effect of Dangui Huoxue Zhitong Granule(Granule of Radix Salviae Miltiorrhizae and Ramulus Cinnamomi for activating blood circulation to stop pain)on pain,inflammation and injury of soft tissues.Methods Animal models of inflammation,pain and traumatic injury of soft tissues were adopted for observing the twisting reaction after intraperitoneal injection of acetic acid,pain threshold of hot-plate test,auricular selling of mice and metatarsal swelling of rats.Results Dangui Huoxue Zhitong Granule could decrease conspicuously the twisting reactions,increase the pain threshold(P