Erratum: Author correction to 'Cardamonin from a medicinal herb protects against LPS-induced septic shock by suppressing NLRP3 inflammasome' Acta Pharmaceutica Sinica B 9 (2019) 734-744

[This corrects the article DOI: 10.1016/j.apsb.2019.02.003.].

Content Determination of 10 Kinds of Chemical Components in Ligusticum chuanxiong under Different Storage Conditions by HPLC / 中国药房

OBJECTIVE： To establish the method for the content determination of chlorogenic acid， ligustrazine， ferulic acid， senkyunolide I， senkyunolide H， coniferly ferulate， senkyunolide A， n-butylphtalide， Z-ligustilide and n-butylidenephthalide in Ligusticum chuanxiong under different storage conditions. METHODS： HPLC method was adopted. The determination was performed on Boston C18 column with mobile phase consisted of acetonitrile-0.5％ acetic acid solution （gradient elution） at the flow rate of 1.0 mL/min. The detection wavelength was set at 285 nm， and the column temperature was 30 ℃. The sample size was 5 μL. RESULTS： The linear range of chlorogenic acid， ligustrazine， ferulic acid， senkyunolide I， senkyunolide H， coniferly ferulate， senkyunolide A， n-butylphtalide， Z-ligustilide n-butylidenephthalide were 0.030 53-0.519 01 μg （r＝0.999 5）， 0.001 02-0.017 34 μg （r＝0.999 9）， 0.012 83-0.218 11 μg （r＝0.999 5）， 0.007 63- 0.129 71 μg （r＝0.999 7）， 0.001 76-0.029 92 μg （r＝0.999 5）， 0.054 74-0.930 58 μg （r＝0.999 9）， 0.215 80-3.668 60 μg （r＝0.999 9）， 0.018 02-0.306 34 μg （r＝0.999 7）， 0.232 50- 3.952 50 μg （r＝0.999 9）， 0.002 40-0.040 80 μg （r＝0.999 5）.The limits of quantitation were 2.073 7， 0.556 6， 0.753 8， 0.231 5， 0.306 9， 0.925 2， 2.295 3， 4.624 0， 3.215 3， 0.910 5 ng， respectively. The detection limits were 0.622 1， 0.167 0， 0.226 1， 0.069 4， 0.092 1， 0.277 6， 0.688 6， 1.387 2， 0.964 6， 0.273 1 ng， respectively. RSD of precision， stability and repeatability tests were all less than 5％ （n＝6）. The recovery rates were 95.90％-103.28％ （RSD＝2.99％， n＝6）， 88.24％-107.84％ （RSD＝4.89％， n＝6）， 95.06％-102.08％ （RSD＝3.97％， n＝6）， 93.67％-101.05％ （RSD＝1.02％， n＝6）， 94.81％-104.33％ （RSD＝2.34％，n＝6）， 94.41％-105.59％ （RSD＝4.32％， n＝6）， 92.76％-104.83％ （RSD＝1.95％， n＝6）， 87.22％-102.56％ （RSD＝2.89％， n＝6）， 94.04％-99.52％ （RSD＝0.92％， n＝6）， 88.51％-103.83％ （RSD＝4.89％， n＝6）， respectively. At 5 ℃ and 15 ℃， no obvious deterioration was observed in medicinal materials. At room temperature， some samples were moth-eaten and mildewed. The content ranges of 6 batches of samples were 0.047 7％-0.160 8％， 0.006 1％- 0.022 7％， 0.048 2％-0.172 2％， 0.023 3％-0.145 2％， 0.004 6％-0.030 7％， 0.085 2％-0.835 4％， 0.182 6％-2.112 7％， 0.009 9％- 0.098 3％， 0.614 9％-3.176 2％ and 0.005 7％-0.036 9％， showing decreasing trend； the decrease rate was in descending order 5 ℃＜15 ℃＜room temperature； at the same storage temperature， the decrease rate of polyethylene plastic bag was lower than that of polypropylene woven bag. CONCLUSIONS： This method is accurate and feasible， and can be used for simultaneous determination of 10 kinds of components in L. chuanxiong. It is suggested that L. chuanxiong medicinal materials should be sealed and packed in dry and cool places and should not be stored for a long time.

Cardamonin from a medicinal herb protects against LPS-induced septic shock by suppressing NLRP3 inflammasome

Aberrant activation of NLRP3 inflammasome has been implicated in the pathogenesis of diverse inflammation-related diseases, and pharmacological molecules targeting NLRP3 inflammasome are of considerable value to identifying potential therapeutic interventions. Cardamonin (CDN), the major active ingredient of the traditional Chinese medicinal herb , has exerted an excellent anti-inflammatory activity, but the mechanism underlying this role is not fully understood. Here, we show that CDN blocks canonical and noncanonical NLRP3 inflammasome activation triggered by multiple stimuli. Moreover, the suppression of CDN on inflammasome activation is specific to NLRP3, not to NLRC4 or AIM2 inflammasome. Besides, the inhibitory effect is not dependent on the expression of NF-B-mediated inflammasome precursor proteins. We also demonstrate that CDN suppresses the NLRP3 inflammasome through blocking ASC oligomerization and speckle formation in a dose-dependent manner. Importantly, CDN improves the survival of mice suffering from lethal septic shock and attenuates IL-1 production induced by LPS , which is shown to be NLRP3 dependent. In conclusion, our results identify CDN as a broad-spectrum and specific inhibitor of NLRP3 inflammasome and a candidate therapeutic drug for treating NLRP3 inflammasome-driven diseases.

Effects of Zhige Oral Liquid on Lipid Metabolism in Rats with Alcoholic Liver Disease / 世界科学技术-中医药现代化

Objective: To observe the effects of Zhige oral liquid on lipid metabolism in rats with alcoholic liver disease.Methods: Alcoholic liver disease rats were induced by alcoholic gavage plus normal diet. The intervention group was given different doses of Zhige oral liquid at the same time. The control group was given alcohol and oral administration of Jiejiuling oral liquid. After 12 weeks, the rats were sacrificed and serum ALT, AST, TC, TG and liver TC and TG levels were measured. Results: Compared with the normal group, the liver index, serum ALT, AST, TC, TG and liver TC and TG levels were significantly higher in the groups except the high dose group (P ＜ 0.01 or 0.05), among which the model group and the low dose group had the most significant increase (P ＜ 0.01) . Compared with the model group, the above indicators were decreased in the high, medium and low dose groups and the control group, and the decreasing trend was the same.That is, the high dose of the Zhige oral liquid was the most significant (P ＜ 0.01), followed by the middle dose group and the control group (P ＜ 0.05) . The decrease in the low dose group was the least significant. Compared with the control group, there were significant differences in the high-dose groups (P ＜ 0.05) . Conclusion: A certain dose of Zhige oral liquid can inhibit or reduce the occurrence of alcoholic liver disease by improving lipid metabolism disorder.

Separation and Identification of Surface Preponderant Fungi on Citri Reticulatae Pericarpium and Its Influence on Effective Substance / 世界科学技术-中医药现代化

This paper studied the influence ofsurface preponderantfungi on main effective substances of CitriReticulatae Pericarpium (CRP).Spreading plate method was used to isolatepredominant strains from the surface of CRP.Besides,the method combining microscopic and molecular identification was also adopted.HPLC and UVspectrophotometric methods were used to determine the main effective substances in CRP.From thesurface of CRP,the advantage strain fungi were Aspergillusniger and A.Flavus.After the inoculation of A.Niger and A.flavus against CRP,the effective compositionwas changed.And different A.niger strains had differenceeffectiveness oneffective chemical components,especially one strain of A.niger.Compared withthe control group,contents of total flavonoids and hesperidin were significantly increased (P＜0.01);and five types of obvious new chemical compositions were produced.It was concluded that the metabolic transformation of fungi was related tochanges ofeffective substances of CRP,which played a significant role in the aging process of CRP.The growth and metabolism of fungi consumeeffeetive substances and producechanges of composition.From the perspective of microorganisms,"the older,the better" of CRP hasa better explanation.

Research Progress on Fungi and Mycotoxin Infection of Chinese Medicinal Materials / 世界科学技术-中医药现代化

Chinese medicinal materials are likely to be polluted by various kinds of fungal toxins during the plantation,collection,transportation and storage,which results in the possibility that quality,safety and treatment effect fails to meet the requirement of safe medicine.This paper reviewed the domestic and overseas studies on the pollution of fungus and fungal toxins in herbs,and made an introduction to the limit standard of fungus and fungal toxins.This paper discussed the importance to establish the inspection standard of various fungal toxins in Chinese medicinal materials and their limit standards to provide references to medication safety and improvement of quality control standard system of Chinese medicinal materials.

The role of suppression of p38 MAPK in cellular vacuole formation / 重庆医学

Objective To investigate the role of the p38 MAPK pathway in the formation of cytoplasmic vacuoles .Methods Af-ter treated with Anisomycin ,SB203580 or SP600125 ,images of HepG2 ,LM3 ,QBC939 ,Hela and A549 cells were recorded by light microscopy and taken at a magnification of 400 × .The effects of anisomycin ,SB203580 and SP600125 on the activity of p38 and JNK were measured by Western blot .LM3 and A549 cells were stained with the ER-tracker red and the lyso-tracker red and subjec-ted to confocal microscopy analysis .Results (1)Anisomycin could abolish cytoplasmic vacuolization of HepG2 cells .(2)p38 MAPK activation was responsible for anisomycin-induced cytoplasmic vacuolization abolishment .(3)p38 MAPK blocking initiated cytoplas-mic vacuoles formation in various cancer cell lines .(4)p38 MAPK blocking-induced cytoplasmic vacuoles disrupted the integrity of endoplasmic reticulum .(5)p38 MAPK blocking reversibly induced cytoplasmic vacuoles formation .Conclusion These observations provide direct evidence for a role of p38 MAPK signaling in regulating the formation of cytoplasmic vacuoles .

Comparative pharmacokinetics of berberine, palmatine and jatrorrhizine in rat plasma after oral administration ofrhizoma coptidis and zuojinwan using liquid chromatography-tandem mass spectrometry

A selective and sensitive liquid chromatography-tandem mass Spectrometry [LC-MS/ MS] method was developed for the determination of berberine, palmatine and Jatrorrhizine in rat plasma. Target compounds, together with the internal standard [metronidazole], were extracted from rat plasma samples by protein precipitation with acetonitrile-methanol [1:2, v/v]. Chromatography was carried out using a C[1g] column [150 x 4.6mm, 5microm] under isocratic elution with water [containing 0.3% formic acid]-acetonitrile [30:70, v/v]. The mass spectrometric detection was performed by selected reaction monitoring [SRM] mode via electrospray ionization [ESI] source operating in positive ionization mode. The method was linear over the concentration range of 0.2-100 microg/mL for all components. The intra- and inter-day precision values were less than 14.7% and the deviations were within +/- 9.0%. The validated method was applied to the comparative pharmacokinetic studies of berberine, palmatine and Jatrorrhizine after oral administration ofRhizoma coptidis and Zuojinwan. The results indicated that the pharmacokinetics of berberine, palmatine and Jatrorrhizine were significantly different between different groups

Relative determination approach to the metabolites of protoberberine alkaloids in rat urine by liquid chromatography tandem mass spectrometry for the comparative studies on rhizome coptidis and zuojinwan preparation

The lack of authentic standards has limited the quantitative analysis of herbal drugs in biological samples. The present work demonstrated a practicable strategy for the assay of herbs and their metabolites independent of authentic standards. A liquid chromatography- electrospray ionization-mass spectrometry [LC-ESI-MS] method for the qualitative and quantitative determination of the metabolites after oral administration of Rhizome coptidis and Zuojinwan preparation in rat urine has been developed. Urine samples, extracted with a protein precipitation procedure were separated on a C[18] column using a mixture of water [containing 0.1% formic acid] and acetonitrile [30:70, v/v] as mobile phase. The detection was performed via MS with electrospray ionization interface in positive selected reaction monitoring [SRM] mode. One urine sample after administration was selected as ‹standard›. The method validation was carried out according to a conventional method which was calibrated by authentic standards. The fully validated method was applied to the pharmacokinetic study of 2,9-demethyljateorhizine-3-sulfate, 13-methoxyjateorhizine-3- glucoronide and 6-methyljateorhizine-5-glucoronide in rat urine. The results could provide evidence to explain the combination of Rhizome coptidis and Evodiae fructus in terms of elimination

Study on fingerprint chromatograms of extract of cimicifugae rhizoma / 中国中药杂志

<p><b>OBJECTIVE</b>To establish the fingerprint chromatograms of the extract of Cimicifugae Rhizoma firstly.</p><p><b>METHOD</b>Phenolic acids and triterpenoid saponins were analyzed by HPLC. Hypersil BDS C18 (4.6 mm x 250 mm, 5 microm) column was used, the mobile phase was composed of acetonitrile -0.1% H3PO4 with gradient elution, flow rate was 1.0 mL x min(-1), column temprature was 30 degrees C, and the detection wavelength was set at 316 nm and 210 nm.</p><p><b>RESULT</b>In the fingerprint of phenolic acids, thirteen feature peaks were found and the RSD of relative retention time and relative peak area were all less than 3% in the precision and repeation experiments. The similarity of ten batches of samples were all more than 0.90. In the fingerprint of triterpenoid saponins, fourteen feature peaks were found and the RSD of relative retention time and relative peak area were all less than 4% in the precision and repeation experiments. The similarity of ten batches of samples were all more than 0.90.</p><p><b>CONCLUSION</b>This method is comprehensive, stable, reliable and can be used to evaluate the quality of the extract of Cimicifugae Rhizoma. It has provided a reference to the analysis on pharmacodynamic deferences of Cimicifuga extracts and also laid the foundation for its further development.</p>

Discussion on forming pattern of dao-di herbs Ligusticum chuanxiong / 中国中药杂志

As a famous-region Dao-di Herbs, Ligusticum chuanxiong which mainly grows in the west of the upper reaches of Jinma River in Dujiangyan for a long time. In recent years, the history, species and origin of L. chuanxiong were researched by many scholars. However, the forming pattern of Dao-di herbs of L. chuanxiong has not been reported systematically. Basing on the general principles of the formation of Dao-di herbs, it can be concluded that the forming pattern of L. chuanxiong is the type of two determinants, which are combined both unique ecological environment of genuine regions and advanced cultivation techniques.

Dynamic phosphoproteomics of insulin and epidermal growth factor signaling in mouse hepatocytes / 中国组织工程研究

BACKGROUND: Both epidermal growth factor (EGF) and insulin transfer their signals into cells by two primary signal transduction pathways,including phosphatidylinositol 3-kinase (PI3K) pathway and mitogen activated protein kinases (MAPK) pathway.But they have different physiological functions.OBJECTIVE: To comparatively assay the dynamic behaviors of phosphoproteomes between EGF and insulin signal transductions in mouse hepatocytes and find key signal proteins.DESIGN,TIME AND SETTING: Randomized grouping controlled observation experiment was performed in the laboratory of Molecular Biology,Luzhou Medical College between July 2005 and April 2006.MATERIALS: Hepatocytes were from Kunming mice of closed population.METHODS: The primarily cultured mouse hepatocytes were labeled with 32p isotope and then randomly divided into three groups: control,EGF-stimulated (received 10 μg/L EGF),and insulin-stimulated (received 100 nmol/L insulin) groups.MAIN OUTCOME MEASURES: After mouse hepatocytes were treated with EGF and insulin for 0,5,20,60 and 120 minutes,the dynamic behaviors of phosphoproteomes(I.e,phosphorylated level) between EGF and insulin signal transductions were comparatively analyzed by two-dimensional electrophoresis method.RESULTS: The categories of all phosphorylated proteins between EGF and insulin-stimulated phosphoproteomes had no apparent difference.The dynamic behaviors of phosphoproteomes of most proteins during EGF signal transduction are parallel with those during insulin stimulation,except the dynamic behaviors of 4 proteins are different significantly.CONCLUSION: Aforementioned 4 phosphorylated proteins were most probably the key members that could distinguish between two signal transduction pathways ornetworks,and determined their major physiological functions respectively.

Study on Chemical Constitutes of Ligusticum chuanxiong and Content Determination of Ligustilide / 中国药房

OBJECTIVE:To study the chemical constitutes of Ligusticum chuanxiong and to establish the method for the content determination of ligustilide. METHODS:The compounds were extracted and percolated by ethanol. Then the samples were separated using silica gel and identified by 1H-NMR,13C-NMR data.HPLC was used to assay the contents of ligustilide. RESULTS:Three compounds were isolated and their structures were identified as Z-ligustilide,Z-6,8',7,3'-diligustilide and Z,Z'-6,6',7,3'a-diligustilide. The contents of ligustilide were no less than 0.70%.CONCLUSION:The separation method can be used to prepare high-purity ligustilide reference substance. And the determination method can be used for quality control of L. chuanxiong.

Determination of Vitamin B_1 and Vitamin B_6 in Gengnianling Capsules by HPLC Simultaneously / 中国药房

OBJECTIVE:To determinate vitamin B1 and vitamin B6 in Gengnianling capsules by HPLC simultaneously .METHODS: The separation was performed on Hypersil-ODS C18 column, methanol - sodium hexanesulfonate solution(20 : 80) was used as mobile phase with a flow rate of 0.8ml/ min and detection wavelength of 280nm.RESULTS: Linear correlations with peak area scores were achieved when the sample size of vitamin B1 and vitamin B6 were with a range of 0.884?g-2.652?g (r = 0.9 999) and 0.714?g-2.142?g(r = 0.9 999) .respectively, the average recovery of which were 95.87%(RSD = 0.82%) and 101.96% (RSD = 0.86%), respectively .CONCLUSION: The method is simple, accurate and it can be used for quality control of Gengnianling Capsule.

Identification of variations of gene expression of visceral adipose and renal tissue in type 2 diabetic rats using cDNA representational difference analysis / 中华医学杂志(英文版)

<p><b>OBJECTIVES</b>To identify differences in gene expression in renal and visceral adipose tissue in type 2 diabetic rats using cDNA representational difference analysis (RDA) and to explore the molecular pathogenesis of type 2 diabetes and its chronic vascular complications.</p><p><b>METHODS</b>A rat model of type 2 diabetes was generated by administration of a high fat and calorie diet combined with a low dose of streptozocin (STZ) injected into the tail vein. The difference bands were generated by cDNA representational difference analysis (cDNA RDA). The final difference products were ligated into the pUC-18 vector and sequenced. A bioformatics analysis was performed on the obtained expressed sequence tags (ESTs), and then the expression levels of known and novel genes were verified by semi-quantitative reverse transcription-PCR (RT-PCR). At the same time, full-length cDNA of a novel gene was cloned in silico.</p><p><b>RESULTS</b>The type 2 diabetic rats in this experiment experienced hyperglycemia, lipidemia, lower insulin sensitivity and normal body weight. We obtained 9 novel ESTs and 2 novel genes from renal tissue of rats and 6 novel ESTs and 1 known gene, the rat lipoprotein lipase (LPL) gene from their visceral adipose tissue. The 2 novel genes (RS91 and RS2) from the renal tissue were both very similar to serine (or cysteine) proteinase inhibitor, clade F and eukaryotic translation initiation factor 3 and subunit 5 (EIF-3 epsilon). The expression of both novel genes and the LPL gene were upregulated in renal and visceral adipose tissue of type 2 diabetic and fat-enriched rats. Full-length cDNA of the novel gene RS91 was cloned in silico.</p><p><b>CONCLUSIONS</b>(1) The rat model of type 2 diabetes generated in this study was ideal because the disease in the animals closely mimicked type 2 diabetic patients. (2) cDNA RDA is a flexible, inexpensive, more accurate, sensitive and highly effective technique for identifying differences in gene expression. (3) Six novel ESTs and 1 known gene were obtained from rat visceral adipose tissue. The LPL gene was upregulated in adipose tissue of type 2 diabetic and fat-enriched rats, a result possibly related to the diabetic animals' high fat and calorie diet, lipidemia, insulin resistance (RI) and molecular pathogenesis. (4) Nine novel ESTs and 2 novel genes were obtained from the renal tissue of rat. We believe the 2 novel genes to be the serine proteinase inhibitors clade F and EIF-3 epsilon in rats. The upregulation of the 2 novel genes in renal tissue of type 2 diabetic rats and may have been related to their renal impairment.</p>

Isolation and Identification of Coumarin in Dendrobium denneanum Cultivated in Sichuan and Its Quality Standard Study / 中国药房

OBJECTIVE:To establish the quality creteria of Dendrobium denneanum cultivated in Sichuan.METHODS: The reference substance had been isolated and identified.HPLC and TLC were applied for determination and identification respectively.RESULTS: The coumarin control had been successfully isolated and the TLC spots were clear.The linear range of coumarin was 0.096～0.480 ?g with an average recovery of 99.1%(RSD=1.44%,n=6).CONCLUSION: The results can overall reflect the internal quality of D.denneanum cultivated in Sichuan and provide basis for quality evaluation and development of D.denneanum from Sichuan.

Cloning and studying on the expression and function of thyroid hormone-response protein-1 gene, a novel thyroid hormone-response gene from neonatal rat brain / 中华内分泌代谢杂志

Objective To is ol ate novel thyroid hormone-response genes, to study the characterizations of the ir expressions and to predict their possible functions in neonatal rats. Methods A neonatal rat model with congenital hypothyr oidism was established and cDNA fragments of novel thyroid hormone-response gen es from cerebral cortex of neonatal rats were obtained by fluorescence-labeled DD-PCR analysis, subcloning and sequencing. Complete cDNAs of novel thyroid hor mone-response genes were cloned by the techniques of electronic clone, RT-PCR and sequencing, their expressions regulated by thyroid hormone were confirmed b y Northern blot analysis, their distributions, transcription levels in different tissues and different brain areas were further observed by semiquantitative RT -PCR analysis, and their possible functions were postulated through bioinformat ic techniques. Results A novel complete cDNA of thyroid hormone-response protein-1 (TRP-1) gene is cloned. It is 973 bp in f ull-length (Gene Bank accession no. AF348365), and its transcription was enhanc ed in cerebral cortex in neonatal hypothyroidism rats. The expression of its mRN A was very extensive, but more abundant in brain. Its transcriptional level in d ifferent brain areas was not uniform, much higher in olfactory bulb. Its encodin g protein had some significant domains and motifs. Conclusion TRP-1 gene is a new thyroid hormone-response gene and may play an important role during normal brain development. Its abnormal expression may b e partially responsible for neurological defects in brain arising from thyroid h ormone deficiency during critical period for perinatal rats.

Preparation and Clinical Application of Yousaiqing Suppository / 中国药房

OBJECTIVE： To prepare Yousaiqing suppository for treating ulcerative colitis and observe its clinical therapeutic effect.METHODS： The ingredients of suppository， PASA and berberine HCl， were determined with RP-HPLC and the therapeutic effect was observed in comparison with sulfasalazine suppository.RESULTS： This method of quality control was feasible and the effective rate for ulcerative colitis amounted to 70.7％ .CONCLUSION： The quality control method is simple， reliable and suitable to routine analysis and the therapeutic effect is satisfactory.

Association of single nucleotide polymorphism in glucagon-like peptide-1 receptor gene with type 2 diabetes in Shanghai / 中华内分泌代谢杂志

Objective To study the association of glucagon like peptide 1 receptor (GLP1R) gene polymorphism with type 2 diabetes in Han population in Shanghai. Methods In the study, 360 type 2 diabetic patients and 313 normal control subjects were enrolled. Diabetic patients were further subdivided into insulin treated non obese patients (BMI28, 192 subjects). A single nucleotide polymorphism (SNP) rs 2268657 was genotyped in all the subjects enrolled in the study using allele specific real time PCR and its association with type 2 diabetes was examined. Results The frequencies of AA,AG, GG genotype incontrol group were0.086,0.447, 0.466 respectively, 0.155, 0.375, 0.470 in non obese diabetic patient group respectively, and 0.109, 0.500, 0.391 in obese diabetic patient group respectively. There was significant difference of the frequency of genotype AA between control group and non obese diabetic patient group (OR=1.939, P

Determination of dehydroxypresenegenin in Polygalae tenuifolia by HPLC / 中草药

Object To establish an HPLC method for the determination of dehydroxypresenegenin for the quality control of Polygalae tenuifolia Willd. Methods Dehydroxypresenegenin was separated by ODS column (125 mm ? 4 mm, 5 ?m) with a mobile phase of methanol 0 05% phosphoric acid (74∶26) and detected at a wavelength of 210 nm. Results The calibration curve was linear in the range of 0 585～11 7 ?g,with a correlation coefficient of 0 999 99 . The average recovery was 99 21% (n=5), and RSD=0 25% (n=5). Conclusion The method is simple and accurate, with good repeatability and can be used for the quality control of Polygalae tenuifolia Willd and compound preparations of Radix Polygalae.