OBJECTIVE: To study the terpene constituents from Cunninghamia lanceolata (Lamb.) Hook. METHODS: The ethanol crude extract was fractionalized by using EtOAc, then a series of chromatography methods were used to isolate the terpene compounds and the structures were elucidated by their physicochemical properties and spectroscopic data. RESULTS: Nine terpenes were isolated and identified as 8(17),12E,14-labdatrien-18-al(1),4S,5R,9S,10R-8(17),12,14-labdantrien-18-ol acetate(2),4S,5R,9S,10R-8(17),12,14-labdantrien-18-ol(3),4S,57R,9S,10R-8(17), 12,14-labdantrien-18-oic acid(4),3β-hydroxy-8(17),13E-labdandien-15-oic acid(5), hinokiol(6),8,15-pimaradien-18-oic acid(7),15-nor-14-oxo-8(17),12-labdandien-18-oic acid(8), and 15-nor-14-oxo-8(17),12-labdan-dien-18-ol(9). CONCLUSION: Compounds1,2,5-7 and 9 are isolated from this plant for the first time.

Objective To observe the protective effect of magnesium gluconate on myocardial apoptosis by ischemia reperfusion injury in isolated rat hearts, and study the possible mechanism. Methods The hearts of 48 Sprague-Dawely rats were isolated, linked to Lange-ndorff perfusion apparatus, and randomly divided into 3 equal groups(n = 16 each) : Control group, ischemia/reperfusion (I/R) group and magnesium gheonate group. 8 rats in each group were perfused. Control group was pedused with modified KH buffer for 110min. I/B group was perfuesd with modified KH buffer for 20 min, then exposed to iscbemia for 30 min, and then reperfused with modified KH buffer for 60 min. Magnesium gheonate group was perfumed with modified KH buffer with magnesium gluconate for 20 min, then exposed to isohemia for 30 min and then reperfused with modified KH buffer with magnesium glueonate for 60 min. Lacate dehydrogenase (LDH) and ereatine kinase (CK) in the effluent liquid from the heart were measured after reperfusion. The concentration of Ca2+ and NO in the left ventricle were determined. The other 8 rats in each group were reperfused for 120 minutes as the method described before. After repeffusion, the myoeyte apoptosis was examined by Annexin-V-FITC/PI. After the two experiments the incidence of ventrieular arrhytlunias during reperfusion was assessed. Results Compared with I/R, magnesium glueonate decreased the incidence of ventricular an'hythmias(P <0. 01). The contents of CK and LDH in the effluent liquid from the heart in magnesium glueonate group was lower than that of I/R group (P <0. 01). The contents of Ca2+ and NO in the left ventricle in magnesium gluconate group was decreased than that of I/R group (P <0. 01). The index of myocyte apoptosis were significanfly lower in magnesium glueonate group than that of I/R group (apoptosis index :27.79±1.59 vs 33.61±2.10, P < 0. 01) . Conclusion Magnesium glueonate has protective effect on myocardial isohemia reperfusion injury in rats. The protective effect may be related to decreasing myocyte apoptosis by increasing the content of NO and relieving calcium overload.

OBJECTIVE: To compare different pretreatment methods for determination of 10 metallic elements including Pb, Cd, As etc in Ganoderma extract. METHODS: Microwave digestion, automatic wet digestion and electrical heating wet digestion methods were used to conduct pretreatment of Ganoderma extact, then the contents of Pb, Cd, Hg, As, Cu, Fe, Mn, Zn, Al, Zn and Cr were determined by inductively coupled plasma mass spectrometry (ICP-MS). RESULTS: The contents of metallic elements determined by the three kinds of digestion methods were not statistically different except for Hg (P>0.05). The average recoveries of Hg in the Ganoderma samples were 65.28% and 70.39% when processed by automatic digestion and electrical heating digestion method, respectively, much lower than the value of microwave digestion method (98.31%). The recoveries of other metallic elements ranged from 92.19% to 103. 26% by the three kinds digestion methods, with RSDs less than 5%. CONCLUSION: Automatic digestion and electrical heating digestion methods are unable to meet the requirement for simultaneous determination of the 10 kinds of metallic elements in Ganoderma extract, however, microwave digestion method can. As the microwave digestion method is simple and rapid, so it is more suitable for the determination of heavy metals in Ganoderma. It is suggested to choose appropriate sample digestion method according to specific determination requirement on metallic elements in actual situation.

Objective: To establish microwave digestion-inductively coupled plasma mass spectrometry (ICP-MS) method for the determination of 13 kinds of metals (Pb, Cd, Hg, As, Cu, Fe, Mn, Zn, Ba, Al, Co, Cr, and Ni) in 10 kinds of Chinese materia medica (CMM) injections which are Carthamin yellow for injection, Breviscapine for injection, Urokinase for injection, Lugua Polypeptide for injection, Xuesetong for injection, Shuanghuanglian for injection, Tanreqing Injection, Salvia miltiorrhiza Injection, Yinxing Damo Injection, and Danhong Injection). Methods: HNO3 and H2O2 are as acid digestion system; Samples were conducted pretreatment by microwave digestion instrument; Sc, In, Rh, and Re were used as internal standard elements, 13 kinds of elements were determined by ICP-MS at the same time. Results: Different kinds of the elements had a good linear relationship (r>0.999); The limit of detection was range from 0.008 to 0.069 μg/L, the average recovery rate was range from 93.61% to 100.79%; RSD values of the repeatability and precision were all less than 5%, The As and Cd in 10 kinds of CMM injections were all less than the limit of detection, Pb was ranged from 0.109 to 0.017 mg/kg, Cu was ranged from 0.016 to 0.135 mg/kg, Hg was ranged from 0.005 to 0.018 mg/kg, Fe was ranged from 0.335 to 2.081 mg/kg, Mn was ranged from 0.021 to 0.699 mg/kg, Ba was ranged from 0.009 to 0.028 mg/kg, Zn was ranged from 0.005 to 3.186 mg/kg, Al was ranged from 0.518 to 10.012 mg/kg, Co was ranged from 0.003 to 0.028 mg/kg, Cr was ranged from 0.006 to 0.179 mg/kg, Ni was ranged 0.005 to 1.187 mg/kg, five kinds of heavy metals which were needed to control were safe according to Chinese Pharmacopoeia (2010), the contents of Fe, Mn, Zn, and Al were not too high, but their safe limits were needed to be further in-depth researched and analyzed. Conclusion: This method is simple and fast, and has good accuracy and high precision, so it can be recommended for the determination of metal elements in CMM injections and provide the reference for its quality control.

The report presents a case with Enterobius vermicularis infections in Guiyang City, Guizhou Province, aiming to strengthen the attention to parasitic infections.

OBJECTIVE: Recent investigations have demonstrated that Polygonum perfoliatum L. can protect against chemical liver injury, but the mechanism behind its efficacy is still unclear. Therefore, we studied the pharmacological mechanism at work in P. perfoliatum protection against chemical liver injury. METHODS: To evaluate the activity of P. perfoliatum against chemical liver injury, levels of alanine transaminase, lactic dehydrogenase, aspartate transaminase, superoxide dismutase, glutathione peroxidase and malondialdehyde were measured, alongside histological assessments of the liver, heart and kidney tissue. A nontargeted lipidomics strategy based on ultra-performance liquid chromatography quadrupole-orbitrap high-resolution mass spectrometry method was used to obtain the lipid profiles of mice with chemical liver injury and following treatment with P. perfoliatum; these profiles were used to understand the possible mechanisms behind P. perfoliatum's protective activity. RESULTS: Lipidomic studies indicated that P. perfoliatum protected against chemical liver injury, and the results were consistent between histological and physiological analyses. By comparing the profiles of liver lipids in model and control mice, we found that the levels of 89 lipids were significantly changed. In animals receiving P. perfoliatum treatment, the levels of 8 lipids were significantly improved, relative to the model animals. The results showed that P. perfoliatum extract could effectively reverse the chemical liver injury and significantly improve the abnormal liver lipid metabolism of mice with chemical liver injury, especially glycerophospholipid metabolism. CONCLUSION Regulation of enzyme activity related to the glycerophospholipid metabolism pathway may be involved in the mechanism of P. perfoliatum's protection against liver injury. Please cite this article as: Peng L, Chen HG, Zhou X. Lipidomic investigation of the protective effects of Polygonum perfoliatum against chemical liver injury in mice. J Integr Med. 2023; 21(3): 289-301.
Animals ; Mice ; Polygonum/chemistry* ; Lipidomics ; Liver ; Lipids/pharmacology* ; Glycerophospholipids/pharmacology* ; Chemical and Drug Induced Liver Injury/metabolism*

Aim To investigate the hepatotoxicity of rutecarpine(RUT)by using high-content screening technology.Methods HepG2 cells were exposed to different concentrations of RUT for different time, then cell viability was detected by MTT method.Cell count, nucleus injury, mitochondrial membrane potential(MMP), reactive oxygen species(ROS), internal flow of calcium, cell membrane integrity(DIR)were measured by high-content screening technology.The activation of MAPK, NF-κB and JAKs-STATs was assayed by high-content screening technology.The apoptosis was detected by flow cytometry.Results The viability was significantly reduced by 100 μmol·L-1 RUT(P<0.01)after HepG2 cell exposure to RUT for 24 h, the nuclear area decreased and the nuclear morphology was uneven, and after 48 h, the cell count was significantly reduced(P<0.01), the early apoptosis was detected(P<0.01).After HepG2 cell exposure to RUT for 6 h, the levels of ROS and internal flow of calcium significantly increased(P<0.01), and the cell membrane integrity was obviously damaged(P<0.01).After exposure to 100 μmol·L-1 RUT for 24 h, the phosphorylation of ERK, JNK, STAT3 and p38 significantly increased(P<0.01, P<0.05), but there was no significant change in total protein level.The expression of c-Jun and c-Fos was up-regulated at 3 h(P<0.01), and at 3h time point, the phosphorylation of NF-κB p65 significantly increased(P<0.01), but nuclear translocation was not significant.Conclusions Under certain conditions, RUT shows cytotoxicity on HepG2 cells, and its toxic mechanism is mainly related to injury caused by oxidative stress and inflammatory response.

Aim To investigate the protective effect of baicalin on inflammatory response of human microvascular endothelial cells ( HMECs) induced by lipopo- lysaccharides ( LPS) and its mechanism. Methods LPS was applied to establish inflammatory model on HMECs in this work. HMECs were pretreated with different concentrations of baicalin ( 1. 0 x 10 "6 , 1. 0 x 10 ~7 and 1. 0 x 10 "8 mol • L"1) , and then exposed to LPS. The supernatant was removed and assayed for expression of the adhesion molecule ICAM-1, the inflam- atory mediator IL-6 and MCP-1 by using ELISA reagent kits. The inward flow of calcium ion was observed by fluorescence microscope, and the intracellular calcium ion level was measured by flow cytometry. The fluorescence intensity of nucleus NF-kB p65 was detected by immunoflourescent technique. The nucleus transcriptional activity of NF-kB was measured by the dual lu- ciferase reporter assay system. Moreover, the expression of protein NF-kB p65, phospo-NF-kb p65 ( p p65 ) and Toll like receptor 4 (TLR4 ) were detected by Western blot. Results The internal flow of calcium, the phosphorylation of NF-kB p65 and the transcription activity significantly increased, and the expression of ICAM-1, IL-6 and MCP-1 up-regulated after HMECs were exposed to LPS. Baicalin inhibited the inward flow of calcium ion, the nuclear translocation of NF-kB p65 and the up-regulation of transcriptional activity, decreased the expression of ICAM-1, IL-6 and MCP-1, and down-regulated the expression of NF-kB p65, p- p65 and TLR4 in a dose-dependent manner. Conclusions Baicalin possesses a protective effect on inflammatory response of endothelial cells induced by LPS, and its mechanism may be highly related to the inhibition of the internal flow of calcium and the activation of NF-kB signaling pathway, and thus reduce the level of inflammatory response.

Candida albicans is one of the major causes of invasive fungal infections and a serious opportunistic pathogen in immunocompromised individuals. The antimicrobial peptide AMP-17 has prominent anti-Candida activity, and proteomic analysis revealed significant differences in the expression of cell wall (XOG1) and oxidative stress (SRR1) genes upon the action of AMP-17 on C. albicans, suggesting that AMP-17 may exert anti-C. albicans effects by affecting the expression of XOG1 and SRR1 genes. To further investigate whether XOG1 and SRR1 genes were the targets of AMP-17, C. albicans xog1Δ/Δ and srr1Δ/Δ mutants were constructed using the clustered regulatory interspaced short palindromic repeats-associated protein 9 (CRISPR/Cas9) system. Phenotypic observations revealed that deletion of two genes had no significant effect on C. albicans growth and biofilm formation, whereas XOG1 gene deletion affected in vitro stress response and mycelium formation of C. albicans. Drug sensitivity assay showed that the MIC₈₀ values of AMP-17 against xog1Δ/Δ and srr1Δ/Δ mutants increased from 8 μg/mL (for the wild type C. albicans SC5314) to 16 μg/mL, while the MIC₈₀ values against srr1Δ/Δ: : srr1 revertants decreased to the level of the wild type SC5314. In addition, the ability of AMP-17 to inhibit biofilm formation of both deletion strains was significantly reduced compared to that of wild type SC5314, indicating that the susceptibility of the deletion mutants to AMP-17 was reduced in both the yeast state and during biofilm formation. These results suggest that XOG1 and SRR1 genes are likely two of the potential targets for AMP-17 to exert anti-C. albicans effects, which may facilitate further exploration of the antibacterial mechanism of novel peptide antifungal drugs.
Humans ; Candida albicans ; Antimicrobial Peptides ; Proteomics ; Peptides/pharmacology* ; Transcription Factors/metabolism* ; Antifungal Agents/pharmacology*

OBJECTIVE：To study the effects of chrysophanol on the activa tion of microglia and the expression of inflammatory factors in cerebral ischemia model rats. METHODS ：SD rats were randomly divided into sham operation group ， model group and chrysophanol high ，medium，low dose groups [7.88，3.94，1.97 mg/（kg·d）]，with 20 rats in each group （the number was complemented in cases of death or unsuccessful modeling during modeling process ）. Except for sham operation group ， middle cerebral artery occlusion model was established in other groups by improved thread method. After 2 hours of ischemia ， sham operation group and model group were intraperitoneally injected with 1 mL normal saline ，and each administration group was intraperitoneally injected with 1 mL corresponding drug ，once a day ，for 7 consecutive days. After last medication ，the score of neurological impairment was recorded ；cerebral infarction of rats was observed by TTC staining ，and the percentage of cerebral infarction area was calculated. The expression of Iba- 1 positive cells in ischemic penumbra of rats was observed by immunofluorescence staining. The expression of Notch- 1，TNF-α and ICAM-1 in the ischemic penumbra of rats were detected by Western blotting assay. RESULTS ：In sham operation group ，there was no infarction area in the brain tissue ，and the Iba- 1 positive cells in the ischemic penumbra were few and branched. Compared with sham operation group ，the infarction area of cerebral tissue in rats was obvious in model group ； the 052）number of Iba- 1 positive cells in ischemic penumbra were 〔ZQ2017003〕） increased significantly ，and they were in amoeba or round shape；the neurological impairment score ，the percentage of cerebral infarction area ， relative expression of Notch- 1， TNF-α and ICAM-1 protein in ischemic penumbra were increased significantly （P＜0.05）. Compared with m odel rats ，the infarction area of cerebral tissue in each dose group of chrysophanol was reduced to different extent ；the number of Iba- 1 positive cells in ischemic penumbra was decreased ；neurological impairment score ，the percentage of cerebral infarction area ，relative expression of Notch- 1，TNF-α and ICAM-1 protein were significantly decreased （P＜0.05 or P＜0.01）. CONCLUSIONS ：Chrysophanol has a certain protective effect on the brain tissue of cerebral ischemia model rats ，and can relieve the nerve injury. Its mechanism may be associated with inhibiting the activation of microglia and expression of inflammatory factors mediated by Notch pathway.