1.Multivariate quantitative combined with chemometrics for evaluating the quality of Sophora flavescens from different producing areas
Jiahui CHEN ; Qiong LUO ; Junli ZHAO ; Yan HAI ; Chengdong LIU ; Tuya BAI ; Jun LI ; Yuewu WANG
China Pharmacy 2025;36(19):2404-2408
OBJECTIVE To establish a content determination method for multiple components in Sophora flavescens from different origins and to evaluate its quality by combining with chemometrics. METHODS Thirteen batches (No. K1-K13) of S. flavescens from different origins were selected as test samples. A high-performance liquid chromatography-tandem triple quadrupole mass spectrometry (HPLC-MS/MS) method was established to determine the contents of 12 components, including matrine, oxymatrine, betaine, cytisine, N-methylcytisine, sophoridine, genistein, sophoricoside, sophorone, formononetin, sophorolone Ⅰ and norkurarinone in S. flavescens. Chromatographic separation was performed on a Shim-pack GIST-HP C18 column with a mobile phase consisting of methanol (A) and water containing 0.1% formic acid (B), using gradient elution at a flow rate of 0.25 mL/min, column temperature of 35 ℃, and an injection volume of 3 μL. Mass spectrometry was conducted using an electrospray ionization source with positive and negative ion scanning. Data were collected in segments using the multiple reaction monitoring mode. Technique for order preference by similarity to ideal solution (TOPSIS) and grey relational analysis (GRA)methods were employed to compare and comprehensively evaluate the 13 batches of S. flavescens from different origins. RESULTS The methodological validation for the content determination met the relevant regulatory requirements. The contents of the 12 components were 490.66-1 231.00, 11 088.10- 18 021.50, 7.91-25.38, 903.97-1 713.64, 336.08-1 485.54,1 065.33-2 075.50, 27.52-71.80, 109.36-517.83, 6 034.55-10 632.73, 21.26-145.35, 814.84-1 911.32, 1 040.87-3 446.37 μg/g), respectively. TOPSIS results showed that the top 7 samples in Euclidean distance ranking were K6, K12, K11, K3, K5, K10, K13. The GRA results showed that the top 7 samples in the relative correlation ranking were K12, K11, K10, K6, K13, K5, K3. CONCLUSIONS The established HPLC-MS/MS method is rapid, accurate, highly sensitive, stable and reliable. Combined with chemometrics methods, it can be used for the quality control and evaluation of S. flavescens. The comprehensive quality of samples K3, K5, K6( from Hebei), K10( from Sichuan), K11-K13( from Shanxi), etc. is relatively superior.
2.Exploration of Spectrum-effect Relationship of Zhuriheng Dropping Pills Against Macrophage Foaming Based on UPLC-Q-Exactive Orbitrap MS
Qiong ZHAI ; Fangyuan LIANG ; Yuewu WANG ; Ren BU ; Xin DONG ; Jingkun LU ; Peifeng XUE
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(10):176-186
ObjectiveThrough the correlation analysis between intestinal absorption profile and inhibition of macrophage foaming, the pharmacodynamic components of Zhuriheng dripping pills(ZRH) were explored to provide a basis for establishing its quality standard. MethodIntestinal absorption fluids with 0, 5, 10, 15, 20 times clinical equivalent doses were prepared by a rat everted gut sac(EGS), and the oxidized low density lipoprotein(ox-LDL)-induced RAW264.7 macrophage foaming model was used to investigate the effect of intestinal absorption fluid with different doses on the accumulation of lipids in RAW264.7 cells by oil red O staining and cholesterol content determination, and to screen for the optimal dose. Ultra performance liquid chromatography-quadrupole-electrostatic field orbitrap high-resolution mass spectrometry(UPLC-Q-Exactive Orbitrap MS) was used to analyze and identify intestinal absorption fractions of ZRH intestinal absorption fluids, and partial least squares-discriminant analysis(PLS-DA) and orthogonal partial least squares-discriminant analysis(OPLS-DA) were performed on different doses of ZRH intestinal absorption fluids using SIMCA 13.0 with peak area as the independent variable and the pharmacodynamic indicators as the dependent variables to screen the compounds with variable importance in the projection(VIP) value>1.0 as contributing components, and Pearson correlation analysis was used to determine the spectral effect relationship, determined the compounds and positive correlation with pharmacodynamic were as active ingredients. Molecular docking was used to verify the binding energy of peroxisome proliferator-activated receptor α(PPARα), PPARγ, PPARβ, human retinoid X receptor α(RXRA) and nuclear transcription factor-κB(NF-κB) with the active ingredients in ZRH intestinal absorption fluids. Real-time fluorescence quantitative polymerase chain reaction(Real-time PCR) was performed to detect the mRNA levels of PPARγ, scavenger receptor A1(SRA1) and adenosine triphosphate-binding cassette transporter A1(ABCA1) in RAW264.7 cells, Westen blot was used to detect the expression level of PPARγ protein in RAW264.7 cells, and enzyme-linked immunosorbent assay(ELISA) was used to detect the levels of interleukin(IL)-1β and NF-κB in RAW264.7 cells. ResultAccording to the results of oil red O staining and cholesterol content determination, the ZRH intestinal absorption fluids could significantly reduce macrophage foaming, and intestinal absorption fluids with 15, 20 times clinical equivalent doses had the best effect, the 15-fold ZRH intestinal absorption fluid was finally determined as the study subject. Spectral effect relationship showed that 52 corresponding peaks in the ZRH-containing intestinal fluid were positively correlated with the efficacy, including organic acids, phenylpropanoids, iridoids, flavonoids, bile acids, coumarins and chromones. Target validation results showed that 86.9%-96.2% of the total components processed good binding activities with the key targets of PPARα, PPARγ, PPARβ, RXRA and NF-κB, and the docking energy values were all less than -6.0 kcal·mol-1(1 cal≈4.19 J). The results of validation showed that, compared with the normal group, the model group showed a significant increase in the levels of SRA1 and PPARγ mRNA expression, a significant decrease in ABCA1 mRNA expression, a significant increase in the level of PPARγ protein expression, and a significant increase in the levels of IL-1β and NF-κB(P<0.01), compared with the model group, the 15-fold intestinal absorption fluid group showed a significant decrease in the levels of SRA1 and PPARγ mRNA expression(P<0.05, P<0.01), ABCA1 mRNA expression level was significantly up-regulated, the levels of IL-1β and NF-κB were significantly reduced(P<0.01), and PPARγ protein expression level was significantly reduced(P<0.05). ConclusionThis study identifies 52 components and their metabolites in ZRH intestinal absorption fluid that are positively correlated with the inhibition of macrophage foaming, which may be related to the regulation of the PPARs pathway in cells and the reduction of the levels of inflammatory factors, and can provide a reference for the quality control and clinical application of ZRH.
3.Method establishment of component identification and content determination of 7 components in Mongolian medicine Ixeris chinensis
Lijun SUN ; Jun LI ; Qiutong WANG ; Huimin XIA ; Yuewu WANG ; Qian ZHANG ; Huiwen ZHANG ; Huanyun WANG
China Pharmacy 2023;34(24):3006-3011
OBJECTIVE To establish the methods to identify the chemical components of Ixeris chinensis, and determine the contents of 7 components (chlorogenic acid, luteolin, quercetin, rutin, protocatechuic acid, isochlorogenic acid A, luteoloside). METHODS HPLC-Q-Exactive-MS was used to identify the chemical components of I. chinensis. The contents of 7 components in I. chinensis, including chlorogenic acid, were determined by HPLC-MS/MS. RESULTS A total of 45 components were identified in I. chinensis, including 20 organic acids, 13 flavonoids, 4 fatty acids, 4 amino acids, 3 nucleosides, and 1 coumarin. The linear range of chlorogenic acid, luteolin, quercetin, rutin, protocatechuic acid, isochlorogenic acid A and luteoloside were 503.00- 25 150.00, 42.00-2 100.00, 5.05-252.50, 20.05-1 002.50, 25.10-1 255.00, 750.00-37 500.00, 196.00-9 800.00 ng/mL (r≥0.999 2), respectively. RSDs of precision, stability and reproducibility tests were all less than 3.00% (n=6), and average recovery ranged from 96.72% to 105.84% (all RSD<4.00%, n=6). The contents of 7 components in 3 batches of I. chinensis were 1 145.77- 3 261.25, 23.75-97.90, 0.92-2.12, 1.06-23.18, 9.35-21.85, 833.25-1 045.58, 199.56-1 869.78 μg/g, respectively. CONCLUSIONS The established methods for identification and content determination are rapid and simple, and can be used for the identification of chemical components and the content determination of 7 components in I. chinensis.
4.Rapid analysis and identification of chemical components of ethanol extract from Mongolian medicine Sanzi san by HPLC-Q-Exactive-MS
Jun LI ; Yuxia HU ; Mengdi ZHANG ; Yuewu WANG ; Qian ZHANG ; Gejihu HU ; Feng GAO ; Fuhou CHANG
China Pharmacy 2022;33(11):1348-1354
OBJECTIVE To systematic ally stu dy the chemic al components of ethanol extract from Sanzi san ,and to provide reference for clarifying the pharmacodynamic material basis of the formulation. METHODS HPLC-Q-Exactive-MS technology was adopted. The determination was performed on Shim-pack GIST-HP C 18 column with mobile phase consisted of acetonitrile- 0.1% formic acid aqueous solution for gradient elution at the flow rate of 1 mL/min. The column temperature was 40 ℃,and sample size was 10 μL. Mass spectrometry conditions included the electrospray spray ion source was used for detection in positive and negative ion detection modes. Full MS/dd-MS 2 detection mode was adopted ,the resolution of Full MS was 70 000 and the resolution of dd-MS2 was 17 500. The scanning range was m/z 110-1 200. The ion peaks were identified by comparing with the information of control substances ,literature references and self-built database. RESULTS A total of 64 components were identified in the ethanol extract of Mongolian medicine Sanzi san , including 9 flavonoids,13 iridoids,14 organic acids ,18 tannins,3 triterpenes,3 amino acids and 4 fatty acids. CONCLUSIONS The ethanol extract of Mongolian medicine Sanzi san mainly include iridoids ,tannins and flavonoids ,which might be the pharmacodynamic material basis of Sanzi san.
5. Hypertrophic port-wine stain: a clinicopathological analysis of 24 cases
Qiuyu LIU ; Enchao JIA ; Guiming HU ; Ying WANG ; Yubin GONG ; Dongge LI ; Yuewu ZHAO
Chinese Journal of Pathology 2019;48(11):878-883
Objective:
To investigate the clinical and pathologic features, diagnosis and differential diagnosis of hypertrophic port-wine stain (PWS).
Methods:
Cases of hypertrophic PWS, collected from Henan Provincial People′s Hospital between 2012 and 2018, were retrospectively analyzed for their clinical and pathologic features, immunophenotype and histochemical data, and the relevant literature was reviewed.
Results:
Twenty-four cases of PWS were included in this cohort, located in the head and neck region (20 cases), limbs (2 cases), and trunk (2 cases). The clinical presentations were mainly red or purple-red plaques or slow growing, painless nodules, or thickened and raised above the skin surface. Microscopically, deformed blood vessels showed honeycomb-like, plexiform or cluster-like growth pattern, and diffusely involved the dermis, skin appendages, subcutaneous fat tissue, and deep skeletal muscles; The vascular lumen of the deformed vessels was dilated (≥100 μm in diameter), and in 18 cases the lumen was greater than 400 μm. The superficial dermis mainly contained few deformed capillaries. The deep wall showed thickening of blood vessel wall and fibrous tissue hyperplasia. Elastic fiber and Masson staining indicated abnormal venous vessel, which in some cases contained small amount of abnormal arterioid vessel,without vascular endothelial cell proliferation in all cases. In 24 cases, 19 cases had epidermal atrophy, 6 with vascular chronic inflammation or epidermal ulcer, 4 with capillary hemangioma, 4 with sebaceous gland hyperplasia, 2 with epidermal papillary hyperplasia and 2 with vascular keratomas.
Conclusions
PWS is a common congenital capillary malformation. The number of histologically deformed capillaries is reduced and they usually locate in the superficial part. The deep vascular wall is increased with thick venous malformation, diffusely involving the dermis and deep skeletal muscle. Furthermore, PWS needs to be differentiated from infantile hemangioma, cavernous hemangioma and vascular keratomas.
6.The mediating effect of self-consciousness between peer relationship and social anxiety in HIV-infec-ted children
Ruopeng WANG ; Xiuxian YANG ; Xiaohui QIU ; Zhengxue QIAO ; Lin WANG ; Erying ZHAO ; Wenbo WANG ; Jiawei ZHOU ; Enpeng ZHOU ; Yunjia XIE ; Yuewu CHENG ; Yanjie YANG
Chinese Journal of Behavioral Medicine and Brain Science 2019;28(3):266-269
Objective To explore the relationship among peer relationship,self-consciousness and social anxiety in HIV-infected children. Methods Peer Relationship Scale,Piers-Harris Children's Self-con-cept Scale and Social Anxiety Scale for Children were applied to 126 HIV-infected children. Results The score of peer relationship was 53. 0(45. 0,62. 0),the score of self-concept was 52. 0(42. 0,60. 0),and the score of social anxiety was 6. 0(2. 0,9. 0). Peer relationship was positively correlated with social anxiety( r=0. 196,P=0. 029) and negatively correlated with self-consciousness(r=-0. 628,P<0. 01). Self-conscious-ness was negatively correlated with social anxiety(r=-0. 504,P<0. 01). Bootstrap analysis showed that self-consciousness played a complete mediating role between peer relationship and social anxiety,and the media-ting effect value was 0. 377. Conclusion Peer relationship,self-consciousness and social anxiety are closely related. Furthermore,self-consciousness plays a complete mediating role between peer relationship and social anxiety in HIV-infected children.
7.Hypertrophic port?wine stain: a clinicopathological analysis of 24 cases
Qiuyu LIU ; Enchao JIA ; Guiming HU ; Ying WANG ; Yubin GONG ; Dongge LI ; Yuewu ZHAO
Chinese Journal of Pathology 2019;48(11):878-883
investigate the clinical and pathologic features, diagnosis and differential diagnosis of hypertrophic port?wine stain (PWS). Methods Cases of hypertrophic PWS, collected from Henan Provincial People′s Hospital between 2012 and 2018, were retrospectively analyzed for their clinical and pathologic features, immunophenotype and histochemical data, and the relevant literature was reviewed. Results Twenty?four cases of PWS were included in this cohort, located in the head and neck region (20 cases), limbs (2 cases), and trunk (2 cases). The clinical presentations were mainly red or purple?red plaques or slow growing, painless nodules, or thickened and raised above the skin surface. Microscopically, deformed blood vessels showed honeycomb?like, plexiform or cluster?like growth pattern, and diffusely involved the dermis, skin appendages, subcutaneous fat tissue, and deep skeletal muscles; The vascular lumen of the deformed vessels was dilated (≥100 μm in diameter), and in 18 cases the lumen was greater than 400 μm. The superficial dermis mainly contained few deformed capillaries. The deep wall showed thickening of blood vessel wall and fibrous tissue hyperplasia. Elastic fiber and Masson staining indicated abnormal venous vessel, which in some cases contained small amount of abnormal arterioid vessel, without vascular endothelial cell proliferation in all cases. In 24 cases, 19 cases had epidermal atrophy, 6 with vascular chronic inflammation or epidermal ulcer, 4 with capillary hemangioma, 4 with sebaceous gland hyperplasia, 2 with epidermal papillary hyperplasia and 2 with vascular keratomas. Conclusions PWS is a common congenital capillary malformation. The number of histologically deformed capillaries is reduced and they usually locate in the superficial part. The deep vascular wall is increased with thick venous malformation, diffusely involving the dermis and deep skeletal muscle. Furthermore, PWS needs to be differentiated from infantile hemangioma, cavernous hemangioma and vascular keratomas.
8.Diagnosis and Clinical Analysis of Primary Thyroid Lymphoma.
Yong XIE ; Wenjing LIU ; Yuewu LIU ; Wenze WANG ; Mengyi WANG ; Hongfeng LIU ; Xiaoyi LI ; Weisheng GAO
Acta Academiae Medicinae Sinicae 2017;39(3):377-382
Objective To summarize our experiences in the diagnosis and prognosis of different subtypes of primary thyroid lymphoma (PTL). Methods The clinical data of 27 PTL patients who were treated in our hospital from January 1998 to December 2014 were retrospectively analyzed. The pathological types of these patients included B cell lymphoma unclassifiable (BCLU) (n=5),mucosa-associated lymphiod tissue lymphoma (MALT) (n=9),diffuse large B cell lymphoma (DLBCL) (n=12),and T cell lymphoma (n=1). Results Of all these 27 cases,the most common clinical symptom was painless swelling of the neck (n=21,77.8%). Of 7 patients who had received preoperative fine needle biopsy,lymphoma was suspected in 2 cases (28.6%). Among these 7 cases,the positive rate of suspicious lymphoma was 66.7% in 3 DLBCL patients,0 in 3 MALT patients,and 0 in 1 BCLU paitent. Also,25 patients underwent intraoperative frozen pathological examination,which revealed lymphoma or suspicious lymphoma in 16 cases (64.0%); in these patients,the positive rate was 66.7% for BCLU,77.8% for MALT,58.3% for DLBCL,and 0 for T-cell lymphomas. The overall survival was (89.3±12.4) months,and the overall 5-year survival rate was 61.6%. The estimated survival in symptomatic group was 31.6 months,which was significantly shorter than that in asymptomatic group (97.9 months) (P=0.032). Other factors including age,sex,tumor size,tumor stage,international prognostic index,tracheal compression,lactate dehydrogenase,residual tumor,and pathological type showed no significant effect on survival(all P>0.05). Conclusions DLBCL has the highest fine needle biopsy positive rate,MALT has the highest frozen pathological positive rate,and intraoperative frozen pathology has more malignant results than the preoperative fine needle biopsy in the diagnosis. The accompanying lymphoma symptoms may be an adverse prognostic factor.
9. Clinicopathologic analysis of micronodular thymoma with lymphoid stroma
Xiaoyan WANG ; Mei XU ; Ligang SHI ; Yanzhi DING ; Qiong CHENG ; Yuewu ZHAO
Chinese Journal of Pathology 2017;46(12):837-840
Objective:
To investigate the clinicopathologic features of micronodular thymoma with lymphoid stroma(MNT).
Methods:
Five cases of MNT diagnosed from January 2007 to December 2016 in Henan Provincial People′s Hospital were collected.Hematoxylin-Eosin staining and immunohistochemistry were used to evaluate the histological and immunophenotypic characteristics in 5 MNT cases. Epstein-Barr virus (EBV) status was detected by in situ hybridization for EBV-encoded small RNA (EBER). Polymerase chain reaction was used to detect the rearrangement of immunoglobulin genes.
Results:
Five cases were MNT, including 3 male and 2 female patients, mean aged 59 years (from 43 to 63 years). All patients had ananterior mediastinal mass, with no myasthenia gravis and autoimmune diseases, and underwent surgical resection.Half to ten years follow-up showed no recurrence.Grossly, the tumors were solid in 4 cases, and cystic and solid in 1 case; the border was clear. Histologically, the tumors presented as a distribution of micronodules separated by abundant lymphoid stroma with prominent germinal centers. The nodules were composed of neoplastic spindle, oval cells containing bland, oval nuclei.Immunohistochemical study showed strong positivity of the tumor cells for CKpan, CK19, CK5/6 and p63. Stains for EMA, CD117, calretinin, TTF1 were negative in the tumor cells.Scattered CD3, CD1a, and TdT positive immature T lymphocytes were noted in and around tumor nodules. Many lymphocytes in the stroma, including germinal centers, were positive for CD20.The bcl-2 was also detected in lymphocytes in the stroma, mantle and marginal zone of lymphoid follicles, and in part of tumor cells. Tumor cells and lymphocytes were negative for EBER. Immunoglobulin genes rearrangement analysis showed that B lymphocytes were polyclonal.
Conclusions
MNT is a rare thymoma, which occurs in the elderly and has no obvious symptom. After complete resection, the prognosis is very good. The diagnosis should be based on a combination of clinicopathologic features, and other types of thymoma should be excluded.
10.Compatibility Stability of Salvianolic Acid for Injection Combined with 8 Kinds of Common Solvents
Yan ZHOU ; Dekun LI ; Dazheng ZHOU ; Yuewu YANG ; Aichun JU ; Xiaoyi WANG ; Zhengliang YE
China Pharmacy 2017;28(17):2350-2356
OBJECTIVE:To investigate the compatibility stability of Salvianolic acid for injection (SAFI) combined with 8 kinds of common solvents. METHODS:Referring to package inserts,SAFI was collected and combined with 0.9% Sodium chlo-ride injection,5% Glucose injection,10% Glucose injection,Compound sodium chloride injection,Mannitol injection,Glycerol fructose and sodium chloride injection,Sodium lactate Ringer's injection,Glucose and sodium chloride injection,250 mL each re-spectively,and then sealed in the dark at the room temperature. The appearance of mixtures were observed,pH value,the number of insoluble particles,maximal absorption wavelength and maximal absorbance were detected,and the contents of salvianolic acid B,rosmarinic acid,lithospermic acid and salvianolic acid Y in mixtures were determined by HPLC at 0,1,2,4,8 h after mix-ing. RESULTS:Under above condition,no obvious change was found in appearance or pH values of the mixtures within 8 h. Maxi-mal absorption wavelength ranged 284.5-286.0 nm. After mixed with Mannitol injection,the number of particles≥10μm(1-8 h af-ter mixing)and particles ≥25 μm(4-8 h after mixing)exceeded the scope of Chinese Pharmacopoeia(2015 edition);the maxi-mal absorbance changed significantly(RSD=9.17%,n=5);the relative content of salvianolic acid B,rosmarinic acid,lithosper-mic acid and salvianolic acid Y decreased by more than 10%(RSD=14.65%,6.45%,8.97%,12.49%,n=5);after mixed with Sodium lactate Ringer's injection,the relative content of rosmarinic acid and lithospermic acid changed greatly (RSD=14.57%, 7.28%,n=5);after mixed with 5% Glucose injection(4-8 h after mixing)and Glycerol fructose and sodium chloride injection(8 h after mixing),the relative content of rosmarinic acid were less than 90%(RSD=6.30%,4.86%,n=5);and the number of particles ≥25μm exceeded the scope of phamcopoeia after mixing with Glycerol fructose and sodium injection(0 h). The number of insoluble particles in other mixtures were in line with the standard of pharmacopoeia;maximal absorbance had no significant change(RSD<5%,n=5),and the relative content change of analytes were all less than 10%. CONCLUSIONS:Clinical appli-cation of SAFI combined with Mannitol injection,Sodium lactate Ringe's injection and Glycerol fructose and sodium injecrion should be avoided. After mixed with 5% Glucose injection,SAFI should be used within 4 h. SAFI can be compatible with 0.9%Sodium chloride injection,10% Glucose injection,Compound sodium chloride injection and Glucose and sodium chloride injec-tion.

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