1.Toxicity Attenuation Mechanism on Processing Method for Aconiti Lateralis Radix Praeparata in Guilingji Based on Urine Metabolomics
Jiayun XIN ; Jia CHEN ; Xike XU ; Xingrui QI ; Meixin YANG ; Tiantian LIN ; Huibo LEI ; Xianpeng ZU
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(4):166-174
ObjectiveMetabolomics was used to reveal the mechanism of Aconiti Lateralis Radix Praeparata(ALRP) in attenuating toxicity by processing from the aspects of amino acid metabolism, oxidative stress and energy metabolism by analyzing multiple metabolic pathways. MethodTwenty-four rats were randomly divided into control group, raw group and processed group, 8 rats in each group. The raw and processed group were given with 0.64 g·kg-1 of raw ALRP and processed ALRP respectively every day, the control group was given with an equal amount of normal saline once a day. After continuous administration for 7 days, the urine, serum and heart tissue of rats were collected. Pathological examination of the heart was carried out using hematoxylin-eosin(HE) staining, and the activities of lactate dehydrogenase(LDH) and creatine kinase-MB(CK-MB) in serum and cardiac tissues were detected by microplate assay and immunoinhibition assay. The effects of ALRP on rat heart before and after processing were compared and analyzed. Ultra performance liquid chromatography-quadrupole-time-of-flight mass spectrometry(UPLC-Q-TOF-MS) was used to perform urine metabolomics analysis, and multivariate statistical analysis was used to screen for differential metabolites related to ALRP in attenuating toxicity by processing, and pathway enrichment analysis was carried out to explore the processing mechanism. ResultHE staining showed that no obvious pathological changes were observed in the heart tissue of the control group, while obvious infiltration of inflammatory cells such as plasma cells and granulocytes was observed in the heart tissue of the raw group, indicating that the raw ALRP had strong cardiotoxicity. There was no significant difference in HE staining of heart tissue between the processed group and the control group, indicating that the toxicity of ALRP was significantly reduced after processing. Compared with the control group, the activities of LDH and CK-MB were significantly increased in serum and heart tissue of the raw group, and those were significantly decreased in serum and heart tissue of the processed group, suggesting that the myocardial toxicity of processed ALRP was reduced. A total of 108 endogenous differential metabolites associated with the raw ALRP were screened using multivariate statistical analysis in positive and negative modes, of which 51 differential metabolites were back-regulated by the processed ALRP. Biological analysis of the key regulatory pathways and associated network changes showed that the pathways related to toxicity of ALRP mainly included tryptophan metabolism, arginine and proline metabolism, phenylalanine metabolism, aminoacyl-tRNA biosynthesis, alanine, aspartate and glutamate metabolism, etc. The metabolic pathways related to the attenuation of processed ALRP mainly included aminoacyl-tRNA biosynthesis, tryptophan metabolism, phenylalanine, tyrosine and tryptophan biosynthesis, phenylalanine metabolism and caffeine metabolism. ConclusionThe processing technology of ALRP in Guilingji can significantly attenuate the cardiotoxicity of raw products, the mechanism mainly involves amino acid metabolism, oxidative stress and energy metabolism, which can provide experimental bases for the research related to the mechanism of toxicity reduction of ALRP by processing and its clinical safety applications.
2.Clinical Significance of Peripheral Blood Immune Cell Population Detection in Diagnosis and Treatment of Unexplained Recurrent Spontaneous Abortion
Yang YANG ; Huibo RU ; Kun XU ; Chun WU ; Shusong WANG ; Lirong DU
Journal of Practical Obstetrics and Gynecology 2024;40(9):745-750
Objective:To detect the expression levels of various immune cells in peripheral blood,and to ex-plore their relationship with unexplained recurrent spontaneous abortion(URSA)and its clinical significance in di-agnosis and treatment.Methods:A total of 351 patients with URSA who visited Hebei Reproductive Health Hospi-tal between January 2020 and September 2022 were enrolled as the URSA group,while a control group compri-sing 30 healthy women who had experienced uncomplicated term delivery once and had no history of adverse pregnancy outcomes during the same period was selected.The levels of various immune cell subsets in peripheral blood during the luteal phase were measured and compared between the two groups.Receiver operating charac-teristic(ROC)curve analysis was performed to determine the optimal cutoff values of immune cell levels that could assist in URSA diagnosis and treatment,while logistic regression analysis was conducted to identify factors influ-encing URSA occurrence.Results:The number and percentage of CD3+CD8+cells in the peripheral blood of the URSA group were significantly lower compared to those in the control group(P<0.05).Conversely,the per-centage of NK cells was significantly higher in the URSA group than in the control group(P<0.05).Logistic re-gression analysis revealed that the increase in CD3+CD8+cell count in peripheral blood during the luteal phase was a protective factor of URSA(OR<1,P<0.05).ROC curve analysis determined optimal cutoff values for im-mune cell levels to assist in the diagnosing and treating URSA:702.82 cells/μl for CD3+CD8+cell count,28.39%for CD3+CD8+cell percentage,and 12.33%for NK cell percentage.After using the optimal cutoff value to convert continuous variables into binary variables,Logistic regression analysis showed that a CD3+CD8+cell count<702.82 cells/μl and an NK cell percentage>12.33%were independent risk factors for URSA(OR>1,P<0.05).Conclusions:The decrease of CD3+CD8+cells in peripheral blood during midluteal phase is closely related to the occurrence of URSA.The number of CD3+CD8+cells(702.82 cells/μl)and the percentage of NK cells(12.33%)can be used as reference values for the diagnosis and treatment of URSA.
3.Clinical Significance of Peripheral Blood Immune Cell Population Detection in Diagnosis and Treatment of Unexplained Recurrent Spontaneous Abortion
Yang YANG ; Huibo RU ; Kun XU ; Chun WU ; Shusong WANG ; Lirong DU
Journal of Practical Obstetrics and Gynecology 2024;40(9):745-750
Objective:To detect the expression levels of various immune cells in peripheral blood,and to ex-plore their relationship with unexplained recurrent spontaneous abortion(URSA)and its clinical significance in di-agnosis and treatment.Methods:A total of 351 patients with URSA who visited Hebei Reproductive Health Hospi-tal between January 2020 and September 2022 were enrolled as the URSA group,while a control group compri-sing 30 healthy women who had experienced uncomplicated term delivery once and had no history of adverse pregnancy outcomes during the same period was selected.The levels of various immune cell subsets in peripheral blood during the luteal phase were measured and compared between the two groups.Receiver operating charac-teristic(ROC)curve analysis was performed to determine the optimal cutoff values of immune cell levels that could assist in URSA diagnosis and treatment,while logistic regression analysis was conducted to identify factors influ-encing URSA occurrence.Results:The number and percentage of CD3+CD8+cells in the peripheral blood of the URSA group were significantly lower compared to those in the control group(P<0.05).Conversely,the per-centage of NK cells was significantly higher in the URSA group than in the control group(P<0.05).Logistic re-gression analysis revealed that the increase in CD3+CD8+cell count in peripheral blood during the luteal phase was a protective factor of URSA(OR<1,P<0.05).ROC curve analysis determined optimal cutoff values for im-mune cell levels to assist in the diagnosing and treating URSA:702.82 cells/μl for CD3+CD8+cell count,28.39%for CD3+CD8+cell percentage,and 12.33%for NK cell percentage.After using the optimal cutoff value to convert continuous variables into binary variables,Logistic regression analysis showed that a CD3+CD8+cell count<702.82 cells/μl and an NK cell percentage>12.33%were independent risk factors for URSA(OR>1,P<0.05).Conclusions:The decrease of CD3+CD8+cells in peripheral blood during midluteal phase is closely related to the occurrence of URSA.The number of CD3+CD8+cells(702.82 cells/μl)and the percentage of NK cells(12.33%)can be used as reference values for the diagnosis and treatment of URSA.
4.Clinical Significance of Peripheral Blood Immune Cell Population Detection in Diagnosis and Treatment of Unexplained Recurrent Spontaneous Abortion
Yang YANG ; Huibo RU ; Kun XU ; Chun WU ; Shusong WANG ; Lirong DU
Journal of Practical Obstetrics and Gynecology 2024;40(9):745-750
Objective:To detect the expression levels of various immune cells in peripheral blood,and to ex-plore their relationship with unexplained recurrent spontaneous abortion(URSA)and its clinical significance in di-agnosis and treatment.Methods:A total of 351 patients with URSA who visited Hebei Reproductive Health Hospi-tal between January 2020 and September 2022 were enrolled as the URSA group,while a control group compri-sing 30 healthy women who had experienced uncomplicated term delivery once and had no history of adverse pregnancy outcomes during the same period was selected.The levels of various immune cell subsets in peripheral blood during the luteal phase were measured and compared between the two groups.Receiver operating charac-teristic(ROC)curve analysis was performed to determine the optimal cutoff values of immune cell levels that could assist in URSA diagnosis and treatment,while logistic regression analysis was conducted to identify factors influ-encing URSA occurrence.Results:The number and percentage of CD3+CD8+cells in the peripheral blood of the URSA group were significantly lower compared to those in the control group(P<0.05).Conversely,the per-centage of NK cells was significantly higher in the URSA group than in the control group(P<0.05).Logistic re-gression analysis revealed that the increase in CD3+CD8+cell count in peripheral blood during the luteal phase was a protective factor of URSA(OR<1,P<0.05).ROC curve analysis determined optimal cutoff values for im-mune cell levels to assist in the diagnosing and treating URSA:702.82 cells/μl for CD3+CD8+cell count,28.39%for CD3+CD8+cell percentage,and 12.33%for NK cell percentage.After using the optimal cutoff value to convert continuous variables into binary variables,Logistic regression analysis showed that a CD3+CD8+cell count<702.82 cells/μl and an NK cell percentage>12.33%were independent risk factors for URSA(OR>1,P<0.05).Conclusions:The decrease of CD3+CD8+cells in peripheral blood during midluteal phase is closely related to the occurrence of URSA.The number of CD3+CD8+cells(702.82 cells/μl)and the percentage of NK cells(12.33%)can be used as reference values for the diagnosis and treatment of URSA.
5.Clinical Significance of Peripheral Blood Immune Cell Population Detection in Diagnosis and Treatment of Unexplained Recurrent Spontaneous Abortion
Yang YANG ; Huibo RU ; Kun XU ; Chun WU ; Shusong WANG ; Lirong DU
Journal of Practical Obstetrics and Gynecology 2024;40(9):745-750
Objective:To detect the expression levels of various immune cells in peripheral blood,and to ex-plore their relationship with unexplained recurrent spontaneous abortion(URSA)and its clinical significance in di-agnosis and treatment.Methods:A total of 351 patients with URSA who visited Hebei Reproductive Health Hospi-tal between January 2020 and September 2022 were enrolled as the URSA group,while a control group compri-sing 30 healthy women who had experienced uncomplicated term delivery once and had no history of adverse pregnancy outcomes during the same period was selected.The levels of various immune cell subsets in peripheral blood during the luteal phase were measured and compared between the two groups.Receiver operating charac-teristic(ROC)curve analysis was performed to determine the optimal cutoff values of immune cell levels that could assist in URSA diagnosis and treatment,while logistic regression analysis was conducted to identify factors influ-encing URSA occurrence.Results:The number and percentage of CD3+CD8+cells in the peripheral blood of the URSA group were significantly lower compared to those in the control group(P<0.05).Conversely,the per-centage of NK cells was significantly higher in the URSA group than in the control group(P<0.05).Logistic re-gression analysis revealed that the increase in CD3+CD8+cell count in peripheral blood during the luteal phase was a protective factor of URSA(OR<1,P<0.05).ROC curve analysis determined optimal cutoff values for im-mune cell levels to assist in the diagnosing and treating URSA:702.82 cells/μl for CD3+CD8+cell count,28.39%for CD3+CD8+cell percentage,and 12.33%for NK cell percentage.After using the optimal cutoff value to convert continuous variables into binary variables,Logistic regression analysis showed that a CD3+CD8+cell count<702.82 cells/μl and an NK cell percentage>12.33%were independent risk factors for URSA(OR>1,P<0.05).Conclusions:The decrease of CD3+CD8+cells in peripheral blood during midluteal phase is closely related to the occurrence of URSA.The number of CD3+CD8+cells(702.82 cells/μl)and the percentage of NK cells(12.33%)can be used as reference values for the diagnosis and treatment of URSA.
6.Clinical Significance of Peripheral Blood Immune Cell Population Detection in Diagnosis and Treatment of Unexplained Recurrent Spontaneous Abortion
Yang YANG ; Huibo RU ; Kun XU ; Chun WU ; Shusong WANG ; Lirong DU
Journal of Practical Obstetrics and Gynecology 2024;40(9):745-750
Objective:To detect the expression levels of various immune cells in peripheral blood,and to ex-plore their relationship with unexplained recurrent spontaneous abortion(URSA)and its clinical significance in di-agnosis and treatment.Methods:A total of 351 patients with URSA who visited Hebei Reproductive Health Hospi-tal between January 2020 and September 2022 were enrolled as the URSA group,while a control group compri-sing 30 healthy women who had experienced uncomplicated term delivery once and had no history of adverse pregnancy outcomes during the same period was selected.The levels of various immune cell subsets in peripheral blood during the luteal phase were measured and compared between the two groups.Receiver operating charac-teristic(ROC)curve analysis was performed to determine the optimal cutoff values of immune cell levels that could assist in URSA diagnosis and treatment,while logistic regression analysis was conducted to identify factors influ-encing URSA occurrence.Results:The number and percentage of CD3+CD8+cells in the peripheral blood of the URSA group were significantly lower compared to those in the control group(P<0.05).Conversely,the per-centage of NK cells was significantly higher in the URSA group than in the control group(P<0.05).Logistic re-gression analysis revealed that the increase in CD3+CD8+cell count in peripheral blood during the luteal phase was a protective factor of URSA(OR<1,P<0.05).ROC curve analysis determined optimal cutoff values for im-mune cell levels to assist in the diagnosing and treating URSA:702.82 cells/μl for CD3+CD8+cell count,28.39%for CD3+CD8+cell percentage,and 12.33%for NK cell percentage.After using the optimal cutoff value to convert continuous variables into binary variables,Logistic regression analysis showed that a CD3+CD8+cell count<702.82 cells/μl and an NK cell percentage>12.33%were independent risk factors for URSA(OR>1,P<0.05).Conclusions:The decrease of CD3+CD8+cells in peripheral blood during midluteal phase is closely related to the occurrence of URSA.The number of CD3+CD8+cells(702.82 cells/μl)and the percentage of NK cells(12.33%)can be used as reference values for the diagnosis and treatment of URSA.
7.Clinical Significance of Peripheral Blood Immune Cell Population Detection in Diagnosis and Treatment of Unexplained Recurrent Spontaneous Abortion
Yang YANG ; Huibo RU ; Kun XU ; Chun WU ; Shusong WANG ; Lirong DU
Journal of Practical Obstetrics and Gynecology 2024;40(9):745-750
Objective:To detect the expression levels of various immune cells in peripheral blood,and to ex-plore their relationship with unexplained recurrent spontaneous abortion(URSA)and its clinical significance in di-agnosis and treatment.Methods:A total of 351 patients with URSA who visited Hebei Reproductive Health Hospi-tal between January 2020 and September 2022 were enrolled as the URSA group,while a control group compri-sing 30 healthy women who had experienced uncomplicated term delivery once and had no history of adverse pregnancy outcomes during the same period was selected.The levels of various immune cell subsets in peripheral blood during the luteal phase were measured and compared between the two groups.Receiver operating charac-teristic(ROC)curve analysis was performed to determine the optimal cutoff values of immune cell levels that could assist in URSA diagnosis and treatment,while logistic regression analysis was conducted to identify factors influ-encing URSA occurrence.Results:The number and percentage of CD3+CD8+cells in the peripheral blood of the URSA group were significantly lower compared to those in the control group(P<0.05).Conversely,the per-centage of NK cells was significantly higher in the URSA group than in the control group(P<0.05).Logistic re-gression analysis revealed that the increase in CD3+CD8+cell count in peripheral blood during the luteal phase was a protective factor of URSA(OR<1,P<0.05).ROC curve analysis determined optimal cutoff values for im-mune cell levels to assist in the diagnosing and treating URSA:702.82 cells/μl for CD3+CD8+cell count,28.39%for CD3+CD8+cell percentage,and 12.33%for NK cell percentage.After using the optimal cutoff value to convert continuous variables into binary variables,Logistic regression analysis showed that a CD3+CD8+cell count<702.82 cells/μl and an NK cell percentage>12.33%were independent risk factors for URSA(OR>1,P<0.05).Conclusions:The decrease of CD3+CD8+cells in peripheral blood during midluteal phase is closely related to the occurrence of URSA.The number of CD3+CD8+cells(702.82 cells/μl)and the percentage of NK cells(12.33%)can be used as reference values for the diagnosis and treatment of URSA.
8.Clinical Significance of Peripheral Blood Immune Cell Population Detection in Diagnosis and Treatment of Unexplained Recurrent Spontaneous Abortion
Yang YANG ; Huibo RU ; Kun XU ; Chun WU ; Shusong WANG ; Lirong DU
Journal of Practical Obstetrics and Gynecology 2024;40(9):745-750
Objective:To detect the expression levels of various immune cells in peripheral blood,and to ex-plore their relationship with unexplained recurrent spontaneous abortion(URSA)and its clinical significance in di-agnosis and treatment.Methods:A total of 351 patients with URSA who visited Hebei Reproductive Health Hospi-tal between January 2020 and September 2022 were enrolled as the URSA group,while a control group compri-sing 30 healthy women who had experienced uncomplicated term delivery once and had no history of adverse pregnancy outcomes during the same period was selected.The levels of various immune cell subsets in peripheral blood during the luteal phase were measured and compared between the two groups.Receiver operating charac-teristic(ROC)curve analysis was performed to determine the optimal cutoff values of immune cell levels that could assist in URSA diagnosis and treatment,while logistic regression analysis was conducted to identify factors influ-encing URSA occurrence.Results:The number and percentage of CD3+CD8+cells in the peripheral blood of the URSA group were significantly lower compared to those in the control group(P<0.05).Conversely,the per-centage of NK cells was significantly higher in the URSA group than in the control group(P<0.05).Logistic re-gression analysis revealed that the increase in CD3+CD8+cell count in peripheral blood during the luteal phase was a protective factor of URSA(OR<1,P<0.05).ROC curve analysis determined optimal cutoff values for im-mune cell levels to assist in the diagnosing and treating URSA:702.82 cells/μl for CD3+CD8+cell count,28.39%for CD3+CD8+cell percentage,and 12.33%for NK cell percentage.After using the optimal cutoff value to convert continuous variables into binary variables,Logistic regression analysis showed that a CD3+CD8+cell count<702.82 cells/μl and an NK cell percentage>12.33%were independent risk factors for URSA(OR>1,P<0.05).Conclusions:The decrease of CD3+CD8+cells in peripheral blood during midluteal phase is closely related to the occurrence of URSA.The number of CD3+CD8+cells(702.82 cells/μl)and the percentage of NK cells(12.33%)can be used as reference values for the diagnosis and treatment of URSA.
9.Clinical Significance of Peripheral Blood Immune Cell Population Detection in Diagnosis and Treatment of Unexplained Recurrent Spontaneous Abortion
Yang YANG ; Huibo RU ; Kun XU ; Chun WU ; Shusong WANG ; Lirong DU
Journal of Practical Obstetrics and Gynecology 2024;40(9):745-750
Objective:To detect the expression levels of various immune cells in peripheral blood,and to ex-plore their relationship with unexplained recurrent spontaneous abortion(URSA)and its clinical significance in di-agnosis and treatment.Methods:A total of 351 patients with URSA who visited Hebei Reproductive Health Hospi-tal between January 2020 and September 2022 were enrolled as the URSA group,while a control group compri-sing 30 healthy women who had experienced uncomplicated term delivery once and had no history of adverse pregnancy outcomes during the same period was selected.The levels of various immune cell subsets in peripheral blood during the luteal phase were measured and compared between the two groups.Receiver operating charac-teristic(ROC)curve analysis was performed to determine the optimal cutoff values of immune cell levels that could assist in URSA diagnosis and treatment,while logistic regression analysis was conducted to identify factors influ-encing URSA occurrence.Results:The number and percentage of CD3+CD8+cells in the peripheral blood of the URSA group were significantly lower compared to those in the control group(P<0.05).Conversely,the per-centage of NK cells was significantly higher in the URSA group than in the control group(P<0.05).Logistic re-gression analysis revealed that the increase in CD3+CD8+cell count in peripheral blood during the luteal phase was a protective factor of URSA(OR<1,P<0.05).ROC curve analysis determined optimal cutoff values for im-mune cell levels to assist in the diagnosing and treating URSA:702.82 cells/μl for CD3+CD8+cell count,28.39%for CD3+CD8+cell percentage,and 12.33%for NK cell percentage.After using the optimal cutoff value to convert continuous variables into binary variables,Logistic regression analysis showed that a CD3+CD8+cell count<702.82 cells/μl and an NK cell percentage>12.33%were independent risk factors for URSA(OR>1,P<0.05).Conclusions:The decrease of CD3+CD8+cells in peripheral blood during midluteal phase is closely related to the occurrence of URSA.The number of CD3+CD8+cells(702.82 cells/μl)and the percentage of NK cells(12.33%)can be used as reference values for the diagnosis and treatment of URSA.
10.Determination of 34 prohibited and restricted pesticide residues in Atractylodes by ultra-performance liquid chromatography-tandem mass spectrometry
Chenglin WANG ; Huibo XU ; Xuebin ZHANG ; Haibo GAO ; Zhaokui LI
China Pharmacist 2024;27(7):1125-1133
Objective To establish an ultra-performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS)method for the detection of 34 prohibited and restricted pesticide residues in Atractylodes,and 21 batches of commercially available Atractylodes were detected to preliminarily investigate the pesticide residues in commercial Atractylodes.Methods The samples were extracted with pure acetonitrile,the extracts were purified by Waters-HLB 3cc(60 mg)solid phase extraction column,separated by Waters-C18 column,0.1%formic acid water(containing 10 mmol/L ammonium formate)and acetonitrile were used as mobile phases(gradient elution),and quantitatively analyzed by external standard method in multi-reaction monitoring positive ion mode according to the retention time.Results The linear relationship of each component was good in their respective concentration ranges,the correlation coefficients were all greater than 0.998 0.The limits of detection were within 0.8-4.0 μg/kg,and the limits of quantification were within 2.0-10.0 μg/kg.The average recovery rate was 74.1%-97.4%,and RSD was 0.6%-6.4%(n=9).Seven pesticide residues were found in 13 batches of 21 Atractylodes macrocephalus samples.Conclusion This method is simple to operate,accurate in quantification,has high recovery rate and good repeatability,and is suitable for the detection of multiple residues of prohibited and restricted pesticides in Atractylodes.

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