摘要
Sanhan Huashi formula(SHF) is the intermediate of a newly approved traditional Chinese medicine(TCM) Sanhan Huashi Granules for the treatment of COVID-19 infection. The chemical composition of SHF is complex since it contains 20 single herbal medicines. In this study, UHPLC-Orbitrap Exploris 240 was used to identify the chemical components in SHF and in rat plasma, lung and feces after oral administration of SHF, and heat map was plotted for characterizing the distribution of the chemical components. Chromatographic separation was conducted on a Waters ACQUITY UPLC BEH C_(18)(2.1 mm×100 mm, 1.7 μm) using 0.1% formic acid(A)-acetonitrile(B) as mobile phases in a gradient elution. Electrospray ionization(ESI) source was used to acquire data in positive and negative mode. By reference to quasi-molecular ions and MS/MS fragment ions and in combination with MS spectra of reference substances and compound information in literature reports, 80 components were identified in SHF, including 14 flavonoids, 13 coumarins, 5 lignans, 12 amino-compounds, 6 terpenes and 30 other compounds; 40 chemical components were identified in rat plasma, 27 in lung and 56 in feces. Component identification and characterization of SHF in vitro and in vivo lay foundations for disclosure of its pharmacodynamic substances and elucidation of the scientific connotation.
Subject(s)
Rats , Animals , Tandem Mass Spectrometry , Chromatography, High Pressure Liquid/methods , Drugs, Chinese Herbal/chemistry , COVID-19 , Lignans摘要
Objective To investigate the value of superb micro-vascular imaging (SMI) in evaluating the relationship between neovascularization in carotid atherosclerotic plaque and clinical symptoms.Methods A total of 146 patients with carotid atherosclerotic plaque which were found by ultrasonography were selected from March 2015 to February 2017 in the Central Hospital of Jiading District of Shanghai.The patients were divided into clinical symptom group(n =96) and symptomless group (n =50) according to cerebrovascular events.The neovessels of carotid atherosclerotic plaque was observed and classified by SMI and contrast-enhanced ultrasound (CEUS).The relationship between the degree of neovascularization and clinical symptoms was analyzed by univariate analysis and multivariate analysis.SMI and GEUS were compared by consistency analysis,and the sensitivity and specificity of the two methods were analyzed.Results The average thickness of carotid atherosclerotic plaques in clinical symptoms group and symptomless group was (3.09 ± 0.79) and (2.89 ± 0.49) mm respectively,there was no significant difference between the two groups (P > 0.05).There were 80 (83.33%) patients with neovascularization in carotid atherosclerotic plaque in the clinical symptoms group,and there were 20 (40.00%) patients with neovascularization in carotid atherosclerotic plaque in the symptomless group,the proportion of patients with neovascularization in carotid atherosclerotic plaque in the clinical symptoms group was significantly higher than that in the symptomless group (Z =-6.488,P <0.05).The SMI classification was in good agreement with the CEUS classification in the two groups (P < 0.05).SMI showed the blood flow signal within carotid plaque in 100 cases,including 80 cases in the clinical symptoms group and 20 cases in the symptomless group.SMI showed no blood flow signal within the carotid plaques in 46 cases,including 16 cases in the clinical symptoms group and 30 cases in the symptomless group.GEUS showed the blood flow signal within carotid plaque in 109 cases,including 81 cases in the clinical symptoms group and 28 cases in the symptomless group.GEUS showed no blood flow signal within the carotid plaques in 37 cases,including 15 cases in the clinical symptoms group and 22 cases in the symptomless group.The sensitivity and specificity of SMI predicting the occurrence of clinical symptoms was 83.3% and 40.00% respectively.The sensitivity and specificity of GEUS predicting the occurrence of clinical symptoms was 84.4% and 56.00% respectively.Conclusions SMI can easily detect neovessels in carotid atherosclerotic plaques,and the degree of neovascularization is related to the occurrence of clinical symptoms.SMI technique plays an important role in evaluating the neovascularization in carotid plaques.
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To establish a HPLC-MS/MS quantitative method for analysis of 8 constituents in rat bile. The method was applied in the biliary excretion study after oral administration of Desmodii Styracifolii Herba extract. The rat bile samples were collected during 0-1, 1-2, 2-4, 4-6, 6-8, 8-12, 12-24 h. Diamonsil C₁₈column (4.6 mm×150 mm, 5 μm) was adopted and eluted with methanol and 0.01% acetic acid in a gradient mode. The flow rate was 0.8 mL•min⁻¹, and the column temperature was set at 40 ℃. The detection was carried out by a triple quadrupole linearion trap mass spectrometer in the negative ion mode with an electrospray source. Multiple reactions monitoring (MRM) mode was employed. The calibration curves showed a good linearity, with correlation coefficients (r≥0.991 5) for all of the analytes within the concentration range. The intra-day and inter-day precisions (RSD) were both less than 15%, and the accuracies (RE) ranged from -15% to 15%. The method had a good stability, and was suitable for the content determination of 8 constituents in rat bile. The extraction recoveries of the 8 analytes were more than 63.2%, and the RSD of IS-normalized matrix factors were no more than 15%, which met the requirements for analysis. According to the biliary excretion study, the bile excretion rates of the 8 analytes were relatively low, with great difference among the individuals. The results showed that the established method was simple, selective and specific, thus could be applied in the biliary excretion study of the 8 analytes in rat bile.
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<p><b>OBJECTIVE</b>To investigate the dynamic expression of TPM1 in rat model of hepatic fibrosis and hepatic stellate cells induced by TGFβ1.</p><p><b>METHOD</b>Thirty male SD rats were divided into control group (n = 6) and model group (n = 24). The rat model of hepatic fibrosis was established by intraperitoneal injection of dimethylnitrosamine(DMN). The sera were collected from portal vein and liver tissues were taken from animals 2, 4, 6, 8 weeks HSC-T6 cells were cultured and induced 48 hours by 5 ng/ml TGF-β1. The pathological changes of liver were observed by Hematoxylin-Eosin and Masson Staining. Reverse Transcription-polymerase chain reaction (RT-PCR), immunohistochemistry and Western-blotting were used to determine the mRNA and protein expressions of TPM1, TGFβ1 and α-SMA in rat models and HSC-T6 cells and the localization of TPM1 in rat models.</p><p><b>RESULTS</b>Rat models of hepatic fibrosis were successfully established. TPM1 was lowly stained in the wall of blood vessels in portal areas in normal livers, in fibrotic livers TPM1 was mainly stained along the fibrotic septum. The mRNA and protein expressions of TPM1 and α-SMA in rat models of hepatic fibrosis increased at the week 2 and peaked at week 6, which was statistical significance compared to control group, P < 0.05; TGF-β1 increased at week 2 and it was higher than the levels in other groups at week 4, which was statistical significance compared to control group P < 0.05; Correlation analysis showed that TPM1 positively correlated with α-SMA and TGF-β1, rs = 0.688, rs = 0.692, P < 0.01. In HSC-T6, the mRNA expressions of TPM1 and α-SMA increased after being induced by TGF -beta1. compare with control group, the differences were significant, P less than 0.05.</p><p><b>CONCLUSION</b>TPM1 may be playing an important role in the occurrence and development of liver fibrosis. Maybe it could become a potential therapeutic target for hepatic fibrosis.</p>
Subject(s)
Animals , Male , Rats , Hepatic Stellate Cells , Metabolism , Liver , Pathology , Liver Cirrhosis, Experimental , Metabolism , Pathology , Rats, Sprague-Dawley , Tropomyosin , Metabolism摘要
<p><b>OBJECTIVE</b>To observe the incidence of heparin-induced thrombocytopenia (HIT) in patients received unfractionated heparin (UFH) treatment, and explore the feasibility of monitoring HIT by platelet counts, as well as the significance of HIT-antibody test in HIT diagnosis.</p><p><b>METHODS</b>145 patients received UFH treatment in Vascular Surgery Department were studied. Before and after the UFH treatment, platelet counts, HIT-antibody ELISA test and heparin-induced platelet aggregation (HIPA) were tested.</p><p><b>RESULTS</b>Among the 145 patients, thrombocytopenia occurred in 40 (27.6%) cases, HIT-antibody ELISA test positive in 59 (40.7%) cases, HIPA test positive in 26 (17.9%) cases. The HIT was diagnosed in 24 (16.5%) cases, and heparin-induced thrombocytopenia and thrombosis (HITTS) occurred in 5 (3.4% in all cases, and 20.8% in HIT patients). In HIT patients, 15 patients (62.5%) were thrombocytopenia, HIT-antibody positive and HIPA test positive. Platelet counts in all of the 24 patients recovered to normal or level before UFH treatment in 3-6 days after heparin withdrawal therapy.</p><p><b>CONCLUSION</b>HIT can be early diagnosed by monitoring platelet counts, HIT-antibody ELISA test and HIPA test. Withdrawal of heparin therapy in time and use of alternative anticoagulant, HITTS rate might be expected to decline further.</p>
Subject(s)
Adult , Aged , Female , Humans , Male , Middle Aged , Anticoagulants , Enzyme-Linked Immunosorbent Assay , Heparin , Platelet Aggregation , Platelet Count , Thrombocytopenia , Diagnosis摘要
Stilbene glycoside (TSG) has been shown to have many beneficial properties. It is therefore essential to understand the absorption and metabolism of TSG in detail. We determined the recovery of TSG and its metabolites (TSG sulfate/glucuronides) in rat gastric contents, gastric mucosa, portal vein plasma, celiac arterial plasma, bile, and urine after administration of 15 mg of TSG in 0.5 mL physiological saline or incubation for 20 min in situ in the stomach of rats. Within 20 min, (64.0±9.8)% of the administered TSG disappeared from the stomach) later, TSG was recovered in both free and conjugated forms in plasma and bile, but not in urine. On the other hand, only free TSG was detected in the gastric contents and mucosa; it was also detected in the portal vein plasma as (48.1±3.5)% of the total TSG (all forms of TSG). However, the proportion of free TSG in the celiac arterial plasma and bile decreased to 4%-10%. In addition, the proportion of free TSG to total TSG in the liver microsome incubation mixture after TSG was incubated in liver microsome at 37°C for 30 min was very low [(10.6 ± 2.6)%]. These results indicate that TSG could be quickly absorbed from the rat stomach, conjugated in liver and excreted in bile. Such novel information would be helpful for the use of TSG as a beneficial natural product which may improve its proposed efficacy in preventing chronic diseases.
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OBJECTIVE To evaluate the use of antibiotics in general hospitals of Ningbo. METHODS Totally 4 391 case history records in April,2004 of 12 hospitals were investigated on the use of antibiotics. RESULTS Medicines incomes accounted for 55.16% in total revenue in a hospital,and antibiotics accounted for 32.66% among medicines incomes.Antibiotics using rate was 62.11% in internal medicine departments.The percentage of antibiotics using without evidence was 23.92%,the combined antibiotics using rate was 49.89%.The average duration for antibiotics using was 10.83 days.Antibiotics using rate in surgery departments before operation was 71.00%,during operation was 20.48%,the combined antibiotics using rate during operation was 14.50%,antibiotics using rate after operation was 96.55%.Antibiotics using for treatment accounted for 62.37% and for prevention was 24.17%. CONCLUSIONS Antibiotics using rate is high in hospitals in Ningbo.Income of medicine is also an important part of total revenue in a hospital.We should pay more attention to management on clinical use of antibiotics.
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<p><b>OBJECTIVE</b>To compare the ettect and side-ettect of fluoxetine and combination of fluoxetine and Chinese or Tibetan medicine in treating senile depression in plateau district. Methods Ninety patients with diagnosis of senile depression conformed to CCMD-3 standard, in plateau district of 2260 - 3200 m altitude were randomly divided into three groups and treated with fluoxetine (group A), fluoxetine plus Sanpu Xinnao Xin granule (group B) and fluoxetine plus Xiaoyao pill (group C), respectively, 30 cases in each group. Therapeutic effects were evaluated with Hamilton' s depressive scale (HAMD) and treatment emergent symptom scale (TESS) after 6 weeks treatment.</p><p><b>RESULTS</b>There was no significant difference in the therapeutic effects between the three groups. The adverse reaction in Group B and C was less than that in Group A (P<0.01). Conclusion Sanpu Xinnao Xin granule and Xiaoyao pill can raise the tolerance of patients with senile depression in plateau area against the adverse reaction of fluoxetine.</p>
Subject(s)
Aged , Humans , Middle Aged , Altitude , Antidepressive Agents, Second-Generation , Therapeutic Uses , Depressive Disorder , Drug Therapy , Drug Therapy, Combination , Drugs, Chinese Herbal , Therapeutic Uses , Fluoxetine , Therapeutic Uses , Phytotherapy , Selective Serotonin Reuptake Inhibitors , Therapeutic Uses摘要
<p><b>AIM</b>To study the metabolite of stilbene glucoside in the Chinese traditional medicine Polygonum multiflornm in mice and elucidate its chemical structure by liquid chromatography tandem-mass spectrometry.</p><p><b>METHODS</b>The stilbene glucoside was injected into the tail vein of mice. Blood samples were collected from artery in the eyepit. The methanol-protein-precipitated plasma sample was introduced into the liquid chromatography-tandem mass spectrometer directly. The analytical column was C18 column (250 mm x 4.6 mm ID, 5 microns). The mobile phase consisted of acetonitrile-methanol-water-formic acid (15:18:66:1) for ES+, acetonitrile-methanol-water (15:18:67) for ES-. The UV detection wavelength was set at 320 nm. The mass ion source type was ESI. HV capillary was 3 kV. The dry gas was nitrogen gas and the flow rate was set at 318 L.h-1. The ion source temperature was 150 degrees C.</p><p><b>RESULTS</b>The stilbene glucoside and its metabolite were separated completely under the chromatography condition. The ions at m/z 600 and m/z 605 were detected under positive ion polarity while the ions at m/z 581 and m/z 402 were detected under negative ion polarity.</p><p><b>CONCLUSION</b>It was proposed that the metabolite of stilbene glucoside injected in vein was its glucuronide conjugate.</p>