1.Detection of 10 anti-rheumatic constituents illegally added in Chinese traditional medicine and health products by UPLC-MS/MS
shun Ting LUO ; lan Gui SHI ; yong Jian HU ; hong Ze LIU ; Gang SUN ; Jing YANG
Drug Evaluation Research 2017;40(11):1576-1580
Objective To establish a UPLC-MS/MS method for determination of 10 anti-rheumatic constituents illegally added in Chinese traditional medicine and health products preparation.Methods The column was ACQUITY UPLC BEHC18 (50 mm× 2.1 mm,1.7 μm),The mobile phase was acetonitrile-Ammonium acetate solution (containing 0.1% Acetic acid) with gradient elution at a flow rate of 0.3 mL/min.The ion source was electrospray ionization (ESI),Multiple-reaction monitoring (MRM) was performed to identify and quantify 10 anti-rheumatic constituents.Results 10 linear calibration curves were obtained with r ≥ 0.996 1.The recoveries were determinated at three concentration and ranged from 92.5% to 101.8%.The precision of the method was shown by RSD (n =5) ranged from 0.9% to 3.1%.The ranges of limit of detection were from 0.001 5 to 0.018 μg,and quantitation were from 0.004 5 to 0.55 μg.The illegally added chemicals were detected with 10 batches of 27 batches of samples.Conclusion The method were simple,sensitivity,accurate,and can be used to detect Anti-rheumatic constituents illegally added in Chinese traditional medicine and health products.
2.Differentiated miRNA expression and validation of signaling pathways in apoE gene knockout mice by cross-verification microarray platform.
Hui HAN ; Yu Hong WANG ; Guang Jin QU ; Ting Ting SUN ; Feng Qing LI ; Wei JIANG ; Shan Shun LUO
Experimental & Molecular Medicine 2013;45(3):e13-
The microRNA (miRNA) regulation mechanisms associated with atherosclerosis are largely undocumented. Specific selection and efficient validation of miRNA regulation pathways involved in atherosclerosis development may be better assessed by contemporary microarray platforms applying cross-verification methodology. A screening platform was established using both miRNA and genomic microarrays. Microarray analysis was then simultaneously performed on pooled atherosclerotic aortic tissues from 10 Apolipoprotein E (apoE) knockout mice (apoE-/-) and 10 healthy C57BL/6 (B6) mice. Differentiated miRNAs were screened and cross-verified against an mRNA screen database to explore integrative mRNA-miRNA regulation. Gene set enrichment analysis was conducted to describe the potential pathways regulated by these mRNA-miRNA interactions. High-throughput data analysis of miRNA and genomic microarrays of knockout and healthy control mice revealed 75 differentially expressed miRNAs in apoE-/- mice at a threshold value of 2. The six miRNAs with the greatest differentiation expression were confirmed by real-time quantitative reverse-transcription PCR (qRT-PCR) in atherosclerotic tissues. Significantly enriched pathways, such as the type 2 diabetes mellitus pathway, were observed by a gene-set enrichment analysis. The enriched molecular pathways were confirmed through qRT-PCR evaluation by observing the presence of suppressor of cytokine signaling 3 (SOCS3) and SOCS3-related miRNAs, miR-30a, miR-30e and miR-19b. Cross-verified high-throughput microarrays are optimally accurate and effective screening methods for miRNA regulation profiles associated with atherosclerosis. The identified SOCS3 pathway is a potentially valuable target for future development of targeted miRNA therapies to control atherosclerosis development and progression.
Animals
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Aorta/metabolism/pathology
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Apolipoproteins E/*deficiency/metabolism
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Atherosclerosis/genetics/pathology
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Down-Regulation/genetics
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Gene Expression Profiling
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*Gene Expression Regulation
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Gene Regulatory Networks/genetics
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Immunohistochemistry
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Male
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Mice
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Mice, Inbred C57BL
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Mice, Knockout
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MicroRNAs/*genetics/metabolism
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Models, Biological
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*Oligonucleotide Array Sequence Analysis
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RNA, Messenger/genetics/metabolism
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Reproducibility of Results
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Reverse Transcriptase Polymerase Chain Reaction
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Signal Transduction/*genetics
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Suppressor of Cytokine Signaling Proteins/genetics/metabolism
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Up-Regulation/genetics
3.Establishment of the PCR Amplification and Sequencing System of Hsa-miR-17 Gene Promoter Region
Kai LUO ; dan Meng LIXIE ; Qian WANG ; fang Shun WU ; ting Xiao JIA ; min Zhi HE
Journal of Modern Laboratory Medicine 2017;32(5):4-7,12
Objective To establish the PCR amplification and sequencing system of Hsa-miR-17 gene promoter region.Methods To establish the PCR amplification and sequencing system of Hsa-miR-17 gene promoter region and evaluate its performance by analyzing the sequence characteristics,designing PCR and sequencing primers,extracting sample genome DNA from different human cell lines and using optimization strategy of adding DMSO to the final concentration of 5 % and making use of 5' tailed PCR primers;Then the established system was further verified in 10 other cell line samples.Results The PCR amplification and sequencing system of hsa-miR-17 gene promoter region,which had good repeatability,good specificity and its detection limit was 10 ng/μl,was established successfully under the condition of using 5' tailed PCR primers and 5 % DMSO and that was confirmed by electrophoresis analysis and Sanger sequencing.And fragment of Has-miR-17 gene promoter region in 10 human cell lines were successfully achieved and verified by this system.Conclusion The PCR system of Hsa-miR-17 gene promoter region was established successfully.
4.Juvenile nasopharyngeal angiofibroma: endoscopic surgery and follow-up results.
Bing ZHOU ; Ting CAI ; Qian HUANG ; Xi-hong LIANG ; Xin NI ; Yong-xiang WEI ; Shun-jiu CUI ; Luo ZHANG ; Tong WANG ; Hua-chao LIU ; Ming LIU ; De-min HAN
Chinese Journal of Otorhinolaryngology Head and Neck Surgery 2010;45(3):180-185
OBJECTIVETo present the changes of surgical approaches for the resection of juvenile nasopharyngeal angiofibromas (JNA) and the follow-up results.
METHODSThe clinical records of 59 patients with JNA treated under endoscope between 2002 and 2009 were reviewed retrospectively. The patients were divided into two groups: Group A (Radkowski I a-II b) and Group B (Radkowsik II c-III b). The tumor stages, feeding vessels, operating time, complications and recurrence were observed and recorded.
RESULTSThere were 7 patients with stage I a, 3 patients with stage I b, 5 patients with stage II a, 6 patients with stage II b, 4 patients with stage II c, 23 patients with stage III a, 11 patients with stage III b. The average age at diagnosis in Group A (21 cases) was 23.9 years old. The average operating time was (106.0 +/- 43.7) min. The follow-up ranged from 3 months to 74 months (median 36 months) except for 3 missing cases. The average age at diagnosis in Group B (38 cases) was 16.2 years old and the average operating time was (152.9 +/- 58.0) min. The follow-up ranged from 3 months to 87 months (median 25 months) except for 5 missing cases. During follow-up, 6 patients in group B recurred. Infarction of thalamencephalon happened in 1 patient in group B who recovered totally afterwards. The difference in operating time between two groups was statistically significant (t = -3.232, P = 0.002). The life table was used to evaluate the survival curves and Log-rank test showed that the difference of recurrent time between two groups was statistically significant (P = 0.03).
CONCLUSIONSThe key techniques to remove tumor are bleeding control, drilling-out the bone that tumor invaded. Transpterygoid or posterolateral wall of maxillary sinus approach are recommended for tumors that extend to infratemporal fossa. Small and intermediate-sized JNA which have no deep invasion of skull base (RadkowskiIa-II b) have a low morbidity. JNA at Radkowski stage IIc and above have residual and recurrent risk.
Adolescent ; Adult ; Angiofibroma ; pathology ; surgery ; Child ; Endoscopy ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Nasopharyngeal Neoplasms ; pathology ; surgery ; Neoplasm Recurrence, Local ; Retrospective Studies ; Treatment Outcome ; Young Adult
5.Effect of Compatibility of Anemarrhenae Rhizoma-Phellodendri Chinensis Cortex Couplet Medicines on Glucolipid Metabolism in Type 2 Diabetic Rats Before and After Salt-processing
Shun-ming FAN ; Chun-ling ZHANG ; Jia-qi WANG ; Xing LI ; Ting LUO ; Ling-ying YU
Chinese Journal of Experimental Traditional Medical Formulae 2021;27(3):22-29
Objective:To investigate the effect of compatibility of Anemarrhenae Rhizoma-Phellodendri Chinensis Cortex couplet medicines on glucolipid metabolism in type 2 diabetic rats before and after salt-processing. Method:The type 2 diabetic rat model was induced by high-fat and high-glucose diet combined with low dose streptozotocin (STZ), the model rats were randomly divided into six groups, including the model group, metformin group (200 mg·kg-1), and different compatibility groups of raw and salt-processed of Anemarrhenae Rhizoma and Phellodendri Chinensis Cortex (6.48 g·kg-1). In addition, The same week old rats fed with normal diet were set as the blank group. After 30 d of continuous intragastric administration, changes of fasting blood glucose (FBG), fasting serum insulin (FINS), glycosylated serum protein (GSP), hepatic glycogen, blood lipid [total cholesterol (TC), triglyceride (TG), low density lipoprotein cholesterol (LDL-C), high density lipoprotein cholesterol (HDL-C)], nonesterified fatty acid (NEFA), adipocytokines [adiponectin (ADP) and leptin)], kidney function [blood urea nitrogen (BUN) and creatinine (CRE)] and other indicators of rats from different groups were detected, and the insulin sensitivity index (ISI) and insulin resistance index (HOMA-IR) were calculated, hematoxylin-eosin (HE) staining was used to observe the morphological changes of pancreas, liver and kidney of rats from different groups. Result:Compared with the model group, compatibility of Anemarrhenae Rhizoma-Phellodendri Chinensis Cortex couplet medicines before and after salt-processing all could decrease the levels of FBG, GSP, TC, TG, LDL-C, NEFA, leptin, BUN, CRE and HOMA-IR, and increase the contents of FINS, HDL-C, ADP, hepatic glycogen and ISI, among which the compatibility of salt-processed Anemarrhenae Rhizoma and salt-processed Phellodendri Chinensis Cortex had the most significant effect on regulating glucolipid metabolism in type 2 diabetic rats. The compatibility of all couplet medicines could improve the histopathological changes of pancreas, liver and kidney in type 2 diabetic rats, among which the compatibility of salt-processed Anemarrhenae Rhizoma and salt-processed Phellodendri Chinensis Cortex had the most prominent effect on repairing pathological damage. Conclusion:The compatibility of Anemarrhenae Rhizoma and Phellodendri Chinensis Cortex before and after salt-processing can improve glucolipid metabolism in type 2 diabetic rats, while the comprehensive effect of salt-processed Anemarrhenae Rhizoma and salt-processed Phellodendri Chinensis Cortex
6.Comparative analysis of pharmacokinetics and intestinal absorption of six alkaloids between Sanmiao Pills and Simiao Pills in rats.
Chang-Shun LIU ; Yan-Nan HU ; Zhen-Ye LUO ; Ting XIA ; Xiao-Mei TAN
China Journal of Chinese Materia Medica 2021;46(23):6271-6277
The present study investigated the differences in pharmacokinetics and intestinal absorption of six alkaloids in Sanmiao Pills and Simiao Pills in rats and explored the different efficacies of the two formulae. After oral administration of Sanmiao Pills and Simiao Pills in rats, blood samples were collected at different time points. Samples were prepared for the determination of six alkaloids in plasma by UPLC-MS/MS. The chromatography was performed on an ACE Excel 3 C_(18 )column with acetonitrile-0.1% formic acid in water as the mobile phase for gradient elution. Analytes were detected in the positive ion mode. Plasma concentrations and pharmacokinetic parameters were calculated. Intestinal absorption of alkaloids was investigated by single-pass intestinal perfusion and absorption parameters of ingredients were calculated. The results showed that the UPLC-MS/MS method for simultaneous determination of concentrations of six alkaloids in plasma was developed and validated by methodological investigations, such as specificity, calibration curves, precision, accuracy, recovery, matrix effect, and stability. The results of the pharmacokinetic assay revealed that C_(max) and AUC values of phellodendrine, berberine, magnoflorine, berberrubine, and jatrorrhizine in Simiao Pills were significantly increased, and CL/F values were reduced as compared with those in Sanmiao Pills, which indicated the increase in plasma concentrations of alkaloids. The intestinal absorption parameters K_(a )and P_(eff) values of phellodendrine, berberine, and jatrorrhizine in Simiao Pills were higher than those in Sanmiao Pills. The intestinal absorption and plasma concentrations of alkaloids in Simiao Pills were significantly higher than those in Sanmiao Pills, suggesting that the composition of Simiao Pills was more conducive to the alkaloids into the blood to resist inflammation and lower uric acid.
Alkaloids
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Animals
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Chromatography, High Pressure Liquid
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Chromatography, Liquid
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Drugs, Chinese Herbal
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Intestinal Absorption
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Rats
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Reproducibility of Results
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Tandem Mass Spectrometry
7.Excretion of three alkaloids from Simiao Pills in urine, feces, and bile between normal and type 2 diabetic rats.
Yan-Nan HU ; Zhen-Ye LUO ; Chang-Shun LIU ; Ting XIA ; Feng-Lin ZHANG ; Fei-Long CHEN ; Xiao-Mei TAN
China Journal of Chinese Materia Medica 2023;48(23):6509-6518
This study investigated the differences in excretion kinetics of three alkaloids and their four metabolites from Simiao Pills in normal and type 2 diabetic rats. The diabetes model was established in rats by injection of streptozotocin, and the alkaloids in urine, feces, and bile of normal and diabetic rats were detected by LC-MS/MS to explore the effect of diabetes on alkaloid excretion of Simiao Pills. The results showed that 72 h after intragastric administration of the extract of Simiao Pills, feces were the main excretion route of alkaloids from Simiao Pills. The total excretion rates of magnoflorine and berberine in normal rats were 4.87% and 56.54%, which decreased to 2.35% and 35.53% in diabetic rats, which had statistical significance(P<0.05). The total excretion rates of phellodendrine, magnoflorine, and berberine in the urine of diabetic rats decreased significantly, which were 53.57%, 60.84%, and 52.78% of those in normal rats, respectively. After 12 h of intragastric administration, the excretion rate of berberine in the bile of diabetic rats increased significantly, which was 253.33% of that of normal rats. In the condition of diabetes, the excretion rate of berberine metabolite, thalifendine significantly decreased in urine and feces, but significantly increased in bile. The total excretion rates of jateorrhizine and palmatine in the urine increased significantly, and t_(1/2) and K_e changed significantly. The results showed that diabetes affected the in vivo process of alkaloids from Simiao Pills, reducing their excretion in the form of prototype drug, affecting the biotransformation of berberine, and ultimately increasing the exposure of alkaloids in vivo, which would be conducive to the hypoglycemic effect of alkaloids. This study provides references for the clinical application and drug development of Simiao Pills in diabetes.
Rats
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Animals
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Bile/metabolism*
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Chromatography, Liquid/methods*
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Berberine
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Diabetes Mellitus, Experimental/metabolism*
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Chromatography, High Pressure Liquid/methods*
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Tandem Mass Spectrometry/methods*
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Feces
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Alkaloids/metabolism*
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Diabetes Mellitus, Type 2/metabolism*