Preliminary study on the mechanism of anaphylactoid reaction induced by natural product germacrone / 药学学报

The Chinese medicine injections prepared by the natural products containing sesquiterpenoids caused various adverse reactions in clinical use, among which skin allergic reactions are the most common. However, whether the reason of allergic reaction was related to the three isoprene units contained in the sesquiterpenoids is not clear, so the evaluation of drug safety has important guiding significance. The sesquiterpenoids are small molecular substances, and they are not antigens or haptens. They may induce anaphylaxis reactions by acting mast cells directly. Current research confirmed that Mas-related G protein-coupled receptor-X2 (MRGPRX2) which is a 7-transmembrane G protein coupled receptor on mast cells was a key target mediated allergic reactions induced by many small molecular drugs. Unlike IgE-mediated allergic reactions, pseudo-allergic reaction is related to dosage and dosing rate, and occurs in the first exposure to the sensitizer. In this paper, a series of experiments in vitro found that not all sesquiterpenoids caused anaphylactoid reactions. Ginsenoside Re, ginsenoside Rb1 and germacrone were selected as representative of sesquiterpenoids for calcium imaging assay. The data confirmed that only germacrone activated calcium mobilization through MRGPRX2, causing an increase in intracellular calcium ion concentration in mast cells. Furthermore, the release rate of β-hexosaminidase and the release amount of histamine analysis confirmed that germacrone induced mast cells degranulation directly. Knockdown of MRGPRX2 expression by siRNA and competitive binding experiments against ciprofloxacin were used to prove the target of germacrone was MRGPRX2. The results indicated that germacrone could activate mast cells directly to induce anaphylactoid reaction via MRGPRX2, which might be the reason of skin allergic reactions caused by injections containing germacrone.

Acupuncture price in forty-one metropolitan regions in the United States: An out-of-pocket cost analysis based on OkCopay.com / 中西医结合学报

Few studies have focused on the cost of acupuncture treatments although acupuncture has become popular in the United States (U.S.). The purpose of the current study was to examine the out-of-pocket costs incurred from acupuncture services based on an online website, OkCopay.com. We examined descriptive statistics (range, median and 20% intervals) for the cost of acupuncture "first-time visits" and "follow-up visits" in 41 metropolitan regions in the U.S. The acupuncture prices of 723 clinics throughout 39 metropolitan regions were included, except for Birmingham, Alabama and Detroit, Michigan as there was no online data available at the time of the study for these two regions. The cost range for a first-time acupuncture visit was $15-400; the highest median was $150 in Charleston, South Carolina, while the lowest was $45 in St. Louis, Missouri. The top 10 cities for the highest median were: Baltimore, Washington, D.C., New York, San Francisco, San Jose, Boston, Atlanta, Seattle, Portland and Indianapolis, with the median $120, while the median for all 723 clinics was $112. For the follow-up visits, the cost range was $15-300; the highest median was $108 in Charleston, South Carolina, and the lowest $40 in Miami, Florida. The 10 cities with highest median follow-up acupuncture visit costs were: New York, Baltimore, New Orleans, Washington, D.C., Philadelphia, San Francisco, San Jose, Seattle, Boston and Atlanta, with the median $85, while for all 723 clinics the median price was $80. The estimation of the average gross annual income of each acupuncturist from the regions studied was $95,760, while the total annual cost of patients seeking acupuncture services in the U.S. was about $3.5 billion in 2018.

Effectiveness of two different acupuncture strategies in patients with vulvodynia: Study protocol for a pilot pragmatic controlled trial / 中西医结合学报

<p><b>BACKGROUND</b>Vulvodynia, or vulvar pain, is a common condition in women; however, there are few evidence-based clinical trials evaluating nonpharmacological therapies for this condition. Acupuncture is one complementary and integrative medicine therapy used by some patients with vulvodynia. This study evaluates two different acupuncture strategies for the treatment of vulvodynia and aims to evaluate whether either of the acupuncture protocols reduces vulvar pain, pain duration or pain with intercourse. The study also examines how long the effect of acupuncture lasts in women with vulvodynia.</p><p><b>METHODS/DESIGN</b>The study is designed as a randomized controlled trial, focused on two acupuncture protocols. Fifty-one patients who have had vulvodynia for more than 3 months will be recruited. Among them, 34 patients will be randomized into Groups 1a and 1b; those who are unwilling to receive acupuncture will be recruited into the standard care group (Group 2). Patients in Group 1a will have acupuncture focused on the points in the pudendal nerve distribution area, while patients in Group 1b will receive acupuncture focused on traditional (distal) meridian points. Patients in Group 2 will receive routine conventional treatments, such as using pain medications, local injections and physical therapies or other nonsurgical procedures. Acupuncture will last 45 min per session, once or twice a week for 6 weeks. The primary outcome measurement will be objective pain intensity, using the cotton swab test. The secondary outcome measurement will be subjective patient self-reported pain intensity, which will be conducted before cotton swab test. Pain intensities will be measured by an 11-point Numeric Pain Rating Scale. Pain duration and pain score during intercourse are recorded. Local muscle tension, tenderness and trigger points (Ashi points) are also recorded. All measurements will be recorded at baseline (before the treatment), at the end of each week during treatment and at the end of the 6 weeks. Follow-up will be done 6 weeks following the last treatment.</p><p><b>DISCUSSION</b>Results of this trial will provide preliminary data on whether acupuncture provides better outcomes than nonacupuncture treatments, i.e., standard care, and whether acupuncture focused on the points in pudendal nerve distribution, near the pain area, has better results than traditional acupuncture focused on distal meridian points for vulvodynia.</p><p><b>TRIAL REGISTRATION</b>Clinicaltrials.gov: NCT03481621. Register: March 29, 2018.</p>

Expression of miRNA-21 in colonic mucosa and peripheral blood CD4+ T cells of 49 patients with active inflammatory bowel disease / 中华消化杂志

Objective To investigate the role and significance of miRNA-21 in the pathogenesis and development of inflammatory bowel disease (IBD).Methods The expressions of microRNA-21 (miRNA-21) in colonic mucosa and peripheral blood CD4+T cells were analyzed by fluorescence quantitative polymerase chain reaction (PCR) in 26 patients with active Crohn's disease (CD), 23 patients with active ulcerative colitis (UC) and 19 healthy controls.The relative expression amount of miRNA-21 in colonic mucosal and peripheral blood CD4+ T cells in 11 patients with CD was compared before and after infliximab (IFX) treatment, and its correlation with tumor necrosis factor (TNF)-α was analyzed.t-test was performed for comparison between groups.The correlation between miRNA-21 expression in peripheral blood CD4+ T cells and serum TNF-α was analyzed by Spearman correlation analysis.Results The expression of miRNA-21 in colonic mucosa of CD patients and UC patients was 4.720 ± 0.397 and 4.993±-0.415, respectively, which were both higher than that of healthy control group (1.101±-0.284),and the differences were statistically significant (t =7.657 and 8.356, both P＜0.01).The expression of miRNA-21 in peripheral blood CD4+ T cells of CD patients and UC patients was 3.254 ± 0.323 and 3.450±0.311, which were both higher than that of healthy control group (0.891 ±0.220), and the differences were statistically significant (t=6.007 and 7.098, both P＜0.01).After 12 weeks of IFX treatment, simple endoscopic score for Crohn's disease (SES-CD) of CD patients was 8.019 ± 0.349,which was lower than that before treatment (21.502 ± 1.185), and the difference was statistically significant (t=10.910, P＜0.01).The serum TNF-α level was 61.570±6.252, which was lower than that before treatment (173.500 ± 12.670), and the difference was statistically significant (t=7.926, P＜0.01).The results of correlation analysis indicated that serum TNF-α level was positively correlated with miRNA-21 expression in peripheral blood CD4+ T cells (r =0.780 4, P ＜ 0.01).Conclusions The increasing of miRNA-21 expression in IBD was closely correlated with the serum TNF-α level and severity of inflammation.miRNA-21 might be a new target for IBD treatment.

Professor Qiu Mao-Liang's theory of "three benefits" of acupuncture for diseases of digestive system / 中国针灸

Professor QIU Mao-Iiang is a famous Chinese physician of acupuncture and Chinese medicine in modern time and is one of representative figure of Chengijang acupuncture schools. The theory of "three benefits" of acupuncture for diseases of disgestive system is established according to pathogenesis. He holds that acupuncture is favorable in removing the stagnated liver qi, reinforcing the functions of the spleen and the stomach in treatment of gastroduodenal ulcer. For problems due to spleen deficiency, reinforcing method should be given to the main points, Back-shu points and Yuan-primary points, while reducing method should be given to Front-mu points. Deficiency type of chronic gastritis is much more seen than excessive type. Therefore, the treating principle of acupuncture shoujd be tonifying the spleen and dispeling the cold. Removing the liver qi and regulating qi circulation can be applied in phases. And for special cases, method of tonifying the stomach yin should be used. For gastroptosis, reinforcing qi in the middle jiao should be applied generally. For those with qi stagnation, qi in middle jiao should be regulated. For retention of food and dampness, treatment should focus on removing phlegm and dampness. And the following points can be selected to lift the stomach by long needle, Juque (CV 14), Liangmen (ST 21) and the acupoints around stomach. For treatment of diarrhea, toward the three pathological factors of dampness, heat and cold, treatment of removing dampness, clearing heat and dispelling cold, especially the first one should be applied, and others are selected by differentiation of syndromes. And for gallstone, treating principle should be established according to the theory of qi stagnation, disharmony between the gallbladder and the sto-mach as well as obstruction by substantial pathogens. And regulation of qi, relieving pain, regulating the functions of the gallbladder and the stomach should be applied so as to removing calculus. Professor QIU Mao-Iiang 's theory of "three benefits" of acupuncture for diseases of digestive system is formed on the analysis of the primary and secondary pathologies, the degree of symptoms, progress of the diseases, characteristics of the acupuncture techniques and aims of the treatment. Thus, his theory has great significance in the clinic.

Applications of HPLC/MS in the analysis of traditional Chinese medicines / 药物分析学报

In China, traditional Chinese medicines （TCMs） have been used in clinical applications for thousands of years. The successful hyphenation of high-performance liquid chromatography （HPLC） and mass spectrometry （MS） has been applied widely in TCMs and biological samples analysis. Undoubtedly, HPLC/MS technique has facilitated the understanding of the treatment mechanism of TCMs. We reviewed more than 350 published papers within the last 5 years on HPLC/MS in the analysis of TCMs. The present review focused on the applications of HPLC/MS in the component analysis, metabolites analysis, and pharmacokinetics of TCMs etc. 50% of the literature is related to the component analysis of TCMs, which show that this field is the most popular type of research. In the metabolites analysis, HPLC coupled with electrospray ionization quadrupole time-of-flight tandem mass spectrometry has been demonstrated to be the powerful tool for the characterization of structural features and fragmentation behavior patterns. This paper presented a brief overview of the applications of HPLC/MS in the analysis of TCMs. HPLC/MS in the fingerprint analysis is reviewed elsewhere.

Controlling arachidonic acid metabolic network: from single- to multi-target inhibitors of key enzymes / 药学学报

Inflammatory diseases are common medical conditions seen in disorders of human immune system. There is a great demand for anti-inflammatory drugs. There are major inflammatory mediators in arachidonic acid metabolic network. Several enzymes in this network have been used as key targets for the development of anti-inflammatory drugs. However, specific single-target inhibitors can not sufficiently control the network balance and may cause side effects at the same time. Most inflammation induced diseases come from the complicated coupling of inflammatory cascades involving multiple targets. In order to treat these complicated diseases, drugs that can intervene multi-targets at the same time attracted much attention. The goal of this review is mainly focused on the key enzymes in arachidonic acid metabolic network, such as phospholipase A2, cyclooxygenase, 5-lipoxygenase and eukotriene A4 hydrolase. Advance in single target and multi-targe inhibitors is summarized.

A method for improving the accuracy and sensitivity of cell membrane chromatography / 南方医科大学学报

<p><b>OBJECTIVE</b>To improve the accuracy and sensitivity of cell membrane chromatography (CMC) and evaluate the feasibility of CMC in the study of subtype receptors.</p><p><b>METHODS</b>Plasmids were used to transfer alpha(1B)-AR cDNA into human embryonic kidney 293 (HEK293) cell lines to obtain cell lines stably overexpressing the subtype receptors. HEK293 alpha(1B) cell membrane stationary phase (CMSP) was prepared by immobilizing the cell membrane on silica. The retention time of 9 alpha(1)-adrenoceptor ligands and capacity factors(kappa'(HEK293 alpha1B)) were calculated. The capacity factors of rat liver tissue and primary cultured rat hepatocytes were also calculated for a correlation analysis.</p><p><b>RESULTS</b>The calculated capacity factors (kappa') were positively correlated to the published pKi values. The affinity rank orders were identical. The longest retention of the 9 alpha(1)-adrenoceptor ligands occurred on CMSP prepared with HEK293 alpha(1B) cell lines, while CMSP obtained from rat liver tissue showed the shortest retention of the ligands.</p><p><b>CONCLUSION</b>CMC proves practical in the study of the subtype adrenoceptors. The accuracy and sensitivity of CMC can be improved using HEK293 alpha(1B) cell membrane.</p>

Inhibitory effect of taspine on mouse S180 sarcoma and its mechanism / 中国中药杂志

<p><b>OBJECTIVE</b>To study the inhibition effect of taspine on mouse S180 sarcoma and its mechanism.</p><p><b>METHOD</b>The mouse S180 sarcoma model was established and used to observe the antitumor activity of taspine. The microvessel density and protein expressing of the VEGF, bFGF, Bcl-2 and Bax in the tumor were measured by immunohistochemistry.</p><p><b>RESULT</b>Taspine showed antitumor activity on the mouse S180 sarcoma in a good dose-dependent manner. The inhibition rates on tumor of taspine at low, middle and high concentrations were 39.08% , 43.99% and 48.60%, respectively. The microvessel density and protein expressing of the VEGF, bFGF, Bcl-2 and Bax in the tumor were decreased compared with the negative control. The ratio of Bax to Bcl-2 was increased.</p><p><b>CONCLUSION</b>Taspine has antitumor effect on the S180 sarcoma, and the mechanism may be through the way of decreasing the expressing of the VEGF, bFGF, Bcl-2 and Bax and inducing the vascular endothelial cell apoptosis.</p>

Analysis of supercritical fluid extracts of Radix caulophylli with gas chromatography-mass spectrometry / 药学学报

To analyze the constituents in supercritical fluid CO2 extraction (SFE-CO2) of Radix caulophylli, the Radix caulophylli was extracted with SFE-CO2, and analyzed by gas chromatography-mass spectrometry (GC-MS). The GC-MS analysis with a DB-5MS capillary column (30 mm x 0.32 mm ID, 0.25 microm film thickness) was used. The inlet temperature was maintained at 280 degrees C. The column oven was held at 80 degrees C for 2 min, then programmed from 80 to 280 degrees C at 5 degrees C x min(-1) and, finally, held for 4 min. Helium at a constant flow rate of 2.0 mL x min(-1) was used as the carrier gas. The mass spectrometry conditions were as follows: ionization energy, 70 eV; ion source temperature, 200 degrees C. The mass selective detector was operated in the TIC mode (m/z was from 40 - 500). For the first time 49 peaks were separated and identified, the compounds were quantitatively determined by normalization method, and the identified compounds represent 97.44% of total GC peak areas. Viz, n-hexadecanoic acid (31.4%), (E, E) -9, 12-octadecadienoic acid (26.54%), (Z)-7-tetradecenal (9.4%), hexadecenoic acid (3.23%), 10-undecenal (3.22%), octadecanoic acid (2.25%), and caulophylline (1.76%) etc. The results will provide important foundation for understanding the constituents and further exploitation of Radix caulophylli.

Inhibitory effects of ligustilide and butylidenephthalide on bFGF-stimulated proliferation of rat smooth muscle cells / 药学学报

<p><b>AIM</b>To investigate the bio-affinities of ligustilide and butylidenephthalide to rat aortic smooth muscle cells and the inhibitory effects of them on bFGF-stimulated proliferation of rat vascular smooth muscle cell (VSMC).</p><p><b>METHODS</b>VSMCs were cultured from rat aorta pectoralis and identified by an immunohistochemical method. The bio-affinities between solute (ligustilide or butylidenephthalide) and cell membrane were measured by rat aortic cell membrane chromatography (CMC). The inhibitory effects of ligustilide and butylidenephthalide on bFGF-stimulated VSMC proliferation were evaluated by MIT colorimetric method.</p><p><b>RESULTS</b>Both ligustilide and butylidenephthalide had selective affinities to rat aortic smooth muscle cell as the same as verapamil, one of the calcium ion antagonists. They could potently inhibit the bFGF-stimulated VSMC proliferation at the concentrations of 5.5 and 11.1 micromol x L(-1), separately (P < 0.05), but had no effects on the normal VSMC growth.</p><p><b>CONCLUSION</b>Both ligustilide and butylidenephthalide can inhibit the abnormal proliferation of VSMC induced by bFGF.</p>

Protecting effect of brevifolin and 8,9-single-epoxy brevifolin of Phyllanthus simplex on rat liver injury / 中国中药杂志

<p><b>OBJECTIVE</b>To investigate the effect of protecting liver of brevifolin and 8,9-single-epoxy brevifolin of Phyllanthus simplex.</p><p><b>METHOD</b>Rats were administered with CCl4 (ip) or alcohol (ig) to establish acute or chronic liver injured model, respectively. ALT, AST and TBIL in serum were measured using colorimetric analysis to evaluate liver function. MDA content or SOD activity in serum and liver tissue was measured by thiobarbituric acid chromatometry and xanthine oxidase methods, respectively. The hemorheological parameters were observed.</p><p><b>RESULT</b>Brevifolin and 8,9-single-epoxy brevifolin reduced the increase of ALT induced by CCl4, but they did not influence the increase of AST. And it could inhibit the pathologic increase of serum TBIL induced by alcohol. They could ameliorate the MDA increase or SOD decrease in serum and liver tissue in rats with liver injury, and decrease abnormal changed hemorheological parameters.</p><p><b>CONCLUSION</b>Brevifolin and 8,9-single-epoxy brevifolin show protective effective against acute and chronic liver injuries, and the mechanism is relevant to antagonizing the lipid peroxidation of free radical and improving the blood circulation.</p>

Concentration and determination of strychnine alkaloid in biological fluids / 法医学杂志

OBJECTIVE@#To establish a new method for determination of strychnine alkaloid in biological fluids based on molecularly imprinted polymers.@*METHODS@#A strychnine molecularly imprinted monolithic column was prepared by in-situ molecularly imprinted technique. The polymer was filled to a 1cm column, and a method was developed to concentrate and determine strychnine alkaloids in biological fluids.@*RESULTS@#the limit of detection of the method was 4.9 ng, and the recoveries were more than 92%. The relative standard deviations were smaller than 6.59%. The linear correlation coefficients of standard curves were 0.999 1 and 0.9966 respectively. This method was applied to concentrate and determine strychnine in plasma and urine of poisoned rabbit.@*CONCLUSION@#The new method could concentrate and simultaneously determine strychnine alkaloids in biological fluids, and it was applied to forensic toxicological analysis.

Studies on differences of pharmacokinetic behavior and tissue distribution of nimodipine and its two enantiomers in rats using achiral and chiral liquid chromatography / 药学学报

<p><b>AIM</b>To investigate the differences of pharmacokinetic behavior and tissue distribution of nimodipine and its two enantiomers in rats.</p><p><b>METHODS</b>A high-performance liquid chromatographic method with an ODS column (150 mm x 4.6 mm ID) and a mobile phase of methanol-water (70:30) was used for racemic nimodipine assay. Another method with a Chiralcel OJ column (250 mm x 4.6 mm ID) and a mixture of n-haxane-ethanol (85:15) as mobile phase was used to determine its two enantiomers. Nimodipine was monitored at 236 nm wavelength.</p><p><b>RESULTS</b>The linearity, recoveries and the detection limits of the methods were found to be suitable for the determinations. The average results of within-day and between-day RSDs were 5.64% and 7.85% respectively, the mean recovery was 97.66% for the concentration ranges studied. The pharmacokinetic parameters Tmax, Cmax, AUC and CLs were: S-(-)-nimodipine (2.1 +/- 0.3) h, (197 +/- 5) microgram.L-1, (656 +/- 18) mL.min-1, (0.30 +/- 0.03) microgram.h.L-1, and R-(+)-nimodipine (1.7 +/- 0.5) h, (128 +/- 4) microgram.L-1, (381 +/- 4) mL.min-1, (0.53 +/- 0.03) microgram.h.L-1, respectively. The S-(-)-nimodipine concentration was 2.23 and 1.97 times as high as that of R-(+)-nimodipine in heart and in cerebrum respectively and there was almost only S-(-)-nimodipine in cerebellum. But R-(+)-nimodipine concentration was 1.57, 3.69 and 4.20 times as high as that of S-(-)-nimodipine in major excretion organs such as kidney, spleen and liver respectively.</p><p><b>CONCLUSION</b>The experimental results obtained by using the achiral and chiral liquid chromatography showed that the differences between enantiomers were apparent for the pharmacokinetics in rat plasma, and very significant for the distributions in major target tissues: heart, cerebrum and cerebellum, and main elimination tissues: kidney, spleen and liver.</p>