1.Optimization and Mechanism Exploration of Tusizi Prescription for Ovarian Reserve Function Based on Uniform Design Method
Yuan LI ; Hanqian DU ; Jiashan LI ; Li GUO ; Zehui LI ; Na LIN ; Ying XU
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(7):53-62
ObjectiveTo optimize Tusizi prescription for ovarian reserve function based on the uniform design method combined with in vitro experiments and explore the underlying mechanisms of this prescription. MethodsThe uniform design method was adopted to design a 5-factor 11-level experiment on the water extract of Tusizi prescription. The cell-counting kit-8 (CCK-8) assay was employed to measure the viability of human ovarian granulosa cells (KGN cells) treated with Tusizi prescription extracts 1-11, and multivariate regression analysis was performed to determine the optimal herb ratio in this prescription. The potential targets of active ingredients in the prescription were retrieved from traditional Chinese medicine systems pharmacology database and analysis platform (TCMSP) and encyclopedia of traditional Chinese medicine (ETCM). The common targets shared by Tusizi prescription and diminished ovarian reserve (DOR) were selected and imported into search tool for the retrieval of interacting genes/proteins (STRING) to construct a protein-protein interaction (PPI) network and into gene function annotation database (DAVID) for gene ontology (GO) analysis. The CCK-8 assay was used to measure the viability of ovarian germline stem cells treated with hyperoside. The CCK-8 assay, 5-ethynyl-2'-deoxyuridine (EdU) staining, terminal-deoxynucleoitidyl transferase mediated nick-end labeling (TUNEL), and enzyme-linked immunosorbent assay (ELISA) were employed to examine the proliferation, apoptosis, and estradiol (E2) secretion of KGN cells treated with the water extract 11 of Tusizi prescription (Cuscutae Semen-Lycii Fructus-Dioscoreae Rhizoma-Poria-Nelumbinis Semen 4∶4∶2∶1∶1) and the optimal prescription screened by uniform design. On this basis, the optimal prescription composition for maximizing the effect on ovarian reserve function was determined and preliminary insights into the underlying mechanisms of this prescription were gained. ResultsA total of 147 common targets were obtained from 278 targets of Tusizi prescription and 1 721 targets of DOR. GO analysis revealed 194 biological processes, primarily involving cellular responses to exogenous compound stimuli, negative regulation of apoptotic process, and positive regulation of cell proliferation. It identified 84 cellular components, including cell membrane, mitochondria, and neuronal cell body, as well as 144 molecular functions such as enzyme binding, estrogen response element binding, and nuclear estrogen receptor binding. The multivariate regression analysis revealed that when Tusizi prescription was composed of Cuscutae Semen, Lycii Fructus, Dioscoreae Rhizoma, Poria, and Nelumbinis Semen in a ratio of 27∶30∶17∶12∶14, the water extract of Tusizi prescription had the best effect of enhancing the viability of KGN cells. CCK-8 results showed that compared with the normal group, the hyperoside group demonstrated increased viability of ovarian germline stem cells (P<0.01). The CCK-8, EdU, and ELISA results showed that compared with the normal group, the optimal prescription screened by uniform design and the water extract 11 of Tusizi prescription increased the proliferation and reduced the apoptosis of KGN cells (P<0.05, P<0.01). ELISA results showed that compared with the normal group, the water extract 11 of Tusizi prescription promoted the E2 secretion of KGN cells (P<0.05), while the optimal prescription screened by uniform design had no significant effect on the E2 secretion. ConclusionBoth the water extract 11 of Tusizi prescription (Cuscutae Semen-Lycii Fructus-Dioscoreae Rhizoma-Poria-Nelumbinis Semen 4∶4∶2∶1∶1) and the optimal prescription screened by uniform design (Cuscutae Semen-Lycii Fructus-Dioscoreae Rhizoma-Poria-Nelumbinis Semen 27∶30∶17∶12∶14) can improve the ovarian reserve function, and the former has better effect. Tusizi prescription can modulate biological processes (such as cell proliferation and apoptosis) and molecular functions (such as enzyme binding and estrogen response element binding) through active components like hyperoside to promote the proliferation and E2 secretion and inhibit the apoptosis of KGN cells, thereby protecting the ovarian reserve function.
2.Optimization and Mechanism Exploration of Tusizi Prescription for Ovarian Reserve Function Based on Uniform Design Method
Yuan LI ; Hanqian DU ; Jiashan LI ; Li GUO ; Zehui LI ; Na LIN ; Ying XU
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(7):53-62
ObjectiveTo optimize Tusizi prescription for ovarian reserve function based on the uniform design method combined with in vitro experiments and explore the underlying mechanisms of this prescription. MethodsThe uniform design method was adopted to design a 5-factor 11-level experiment on the water extract of Tusizi prescription. The cell-counting kit-8 (CCK-8) assay was employed to measure the viability of human ovarian granulosa cells (KGN cells) treated with Tusizi prescription extracts 1-11, and multivariate regression analysis was performed to determine the optimal herb ratio in this prescription. The potential targets of active ingredients in the prescription were retrieved from traditional Chinese medicine systems pharmacology database and analysis platform (TCMSP) and encyclopedia of traditional Chinese medicine (ETCM). The common targets shared by Tusizi prescription and diminished ovarian reserve (DOR) were selected and imported into search tool for the retrieval of interacting genes/proteins (STRING) to construct a protein-protein interaction (PPI) network and into gene function annotation database (DAVID) for gene ontology (GO) analysis. The CCK-8 assay was used to measure the viability of ovarian germline stem cells treated with hyperoside. The CCK-8 assay, 5-ethynyl-2'-deoxyuridine (EdU) staining, terminal-deoxynucleoitidyl transferase mediated nick-end labeling (TUNEL), and enzyme-linked immunosorbent assay (ELISA) were employed to examine the proliferation, apoptosis, and estradiol (E2) secretion of KGN cells treated with the water extract 11 of Tusizi prescription (Cuscutae Semen-Lycii Fructus-Dioscoreae Rhizoma-Poria-Nelumbinis Semen 4∶4∶2∶1∶1) and the optimal prescription screened by uniform design. On this basis, the optimal prescription composition for maximizing the effect on ovarian reserve function was determined and preliminary insights into the underlying mechanisms of this prescription were gained. ResultsA total of 147 common targets were obtained from 278 targets of Tusizi prescription and 1 721 targets of DOR. GO analysis revealed 194 biological processes, primarily involving cellular responses to exogenous compound stimuli, negative regulation of apoptotic process, and positive regulation of cell proliferation. It identified 84 cellular components, including cell membrane, mitochondria, and neuronal cell body, as well as 144 molecular functions such as enzyme binding, estrogen response element binding, and nuclear estrogen receptor binding. The multivariate regression analysis revealed that when Tusizi prescription was composed of Cuscutae Semen, Lycii Fructus, Dioscoreae Rhizoma, Poria, and Nelumbinis Semen in a ratio of 27∶30∶17∶12∶14, the water extract of Tusizi prescription had the best effect of enhancing the viability of KGN cells. CCK-8 results showed that compared with the normal group, the hyperoside group demonstrated increased viability of ovarian germline stem cells (P<0.01). The CCK-8, EdU, and ELISA results showed that compared with the normal group, the optimal prescription screened by uniform design and the water extract 11 of Tusizi prescription increased the proliferation and reduced the apoptosis of KGN cells (P<0.05, P<0.01). ELISA results showed that compared with the normal group, the water extract 11 of Tusizi prescription promoted the E2 secretion of KGN cells (P<0.05), while the optimal prescription screened by uniform design had no significant effect on the E2 secretion. ConclusionBoth the water extract 11 of Tusizi prescription (Cuscutae Semen-Lycii Fructus-Dioscoreae Rhizoma-Poria-Nelumbinis Semen 4∶4∶2∶1∶1) and the optimal prescription screened by uniform design (Cuscutae Semen-Lycii Fructus-Dioscoreae Rhizoma-Poria-Nelumbinis Semen 27∶30∶17∶12∶14) can improve the ovarian reserve function, and the former has better effect. Tusizi prescription can modulate biological processes (such as cell proliferation and apoptosis) and molecular functions (such as enzyme binding and estrogen response element binding) through active components like hyperoside to promote the proliferation and E2 secretion and inhibit the apoptosis of KGN cells, thereby protecting the ovarian reserve function.
3.Factors influencing influenza vaccination coverage among kindergarten and primary school children in Zhejiang Province, 2023
Minchao LI ; Jing TAO ; Rui ZHANG ; Yumeng WU ; Zhaokai HE ; Chen WU
Shanghai Journal of Preventive Medicine 2025;37(1):23-28
ObjectiveTo investigate the influenza vaccination coverage among kindergarten and primary school children in Zhejiang Province in 2023 and analyze the influencing factors, and to provide the basis for improving the effect of influenza vaccination in children. MethodsA multi-stage random cluster sampling method was used to select 3 681 parents of children from 10 primary schools and kindergartens based on economic level and geographical distribution in Zhejiang Province, who participated in an online questionnaire survey, including basic information about the children and their parents, parents’ knowledge about influenza, and their willingness to vaccination. ResultsAmong the 3 681 parents surveyed, 33.82% (1 245/3 681) reported that their children received influenza vaccination in 2023. Multivariate logistic regression analysis showed that factors contributing to children’s influenza vaccination included both parents [adjusted OR (95%CI): 1.56 (1.32‒1.84)] and children [6.04 (5.04‒7.27)] having a history of influenza vaccination, parents’ conviction the influenza vaccine could protect children from severe diseases [1.43 (1.19‒1.74)], and the willingness of most parents would let their children get vaccinated [1.40 (1.13‒1.74)]. In contrast, vaccine hesitancy among parents [0.55 (0.43‒0.69)] and the belief that influenza is just a common cold [0.80 (0.65‒1.00)] were hindering factors. ConclusionThe influenza vaccination coverage among children is insufficient. Both the vaccination history of parents and children, as well as parents’ correct understanding of influenza and the effectiveness of influenza vaccine, significantly influence the influenza vaccination status in children. Efforts to address vaccine hesitancy and misconceptions about influenza are essential to improve vaccination rates.
4.Clinical features and genetic analysis of early-onset spinocerebellar ataxia type 5 caused by de novoSPTBN2 gene mutation
Guangjin LUO ; Shuping TANG ; Jiashan LI ; Yang LI ; Chong WANG ; Leihong ZHANG ; Jun CHEN ; Aiyun YUAN ; Mei HOU ; Dianrong SUN
Chinese Journal of Neurology 2024;57(6):607-615
Objective:To summarize the clinical and genetic characteristics of early-onset spinocerebellar ataxia type 5 (SCA5) caused by SPTBN2 gene mutation. Methods:The clinical and genetic data of a child with early-onset SCA5 diagnosed in the Department of Children′s Rehabilitation, Women and Children′s Hospital Affiliated to Qingdao University in February 2022 were retrospectively analyzed. The literatures related to early-onset SCA5 in major databases at home and abroad were retrieved and summarized.Results:The patient, a 4 years and 1 month old girl, was admitted to hospital because of "unable to stand independently at 2 years and 3 months", primarily presented with developmental delay, ataxia, hypotonia, and tendon hyperreflexia during infancy. Progressive cerebellar atrophy was observed on brain magnetic resonance imaging. A de novo heterozygous mutation of the SPTBN2 c.793G>C(p.Asp265His) was identified in the patient. Following hospitalization, the child received comprehensive rehabilitation therapy encompassing physical, occupational, language, educational interventions as well as bicycle ergometer training and transcranial magnetic stimulation. The patient was followed-up for more than 1 year to 4 years and 1 month old, whose motor function, cognitive abilities, and language skills were improved to some extent. A total of 13 English articles and 1 Chinese article were retrieved from the databases. A total of 20 early-onset SCA5 patients have been reported, with onset ages all within 12 months. Infants exhibited decreased muscle tone and delayed motor milestones, with the main clinical manifestations of ataxia, generalized developmental delay, and cerebellar atrophy. The previously reported cases involved 11 mutation sites in the SPTBN2 gene, and the main types of mutations were de novo missense mutations. The mutation site in this case has not been reported in the previous literature. Conclusions:Early-onset SCA5 is a rare autosomal dominant disorder caused by heterozygous mutations in the SPTBN2 gene. The main clinical manifestations include ataxia from infancy, developmental retardation and cerebellar atrophy. Early rehabilitation intervention can improve the degree of the dysfunction.
5.Long-term outcomes of lateral femoral notch after early anterior cruciate ligament reconstruction.
Ming LI ; Wu CAO ; Jun CUI ; Qi-Liang LOU ; Guang-Chen SUN
China Journal of Orthopaedics and Traumatology 2023;36(5):459-464
OBJECTIVE:
To investigate the outcome of lateral femoral notch (LFN) after early anterior cruciate ligament (ACL) reconstruction and evaluate the recovery of knee joint function after the operation.
METHODS:
The clinical data of 32 patients who underwent early ACL reconstruction from December 2015 to December 2019 were retrospectively analyzed. The study included 18 males and 14 females, aged 16 to 54 years old, with an average age of (25.39±2.82) years. The body mass index (BMI) of the patients ranged from 20 to 30 kg/cm2, with an average of (26.15±3.09) kg/cm2. Among them, 6 cases were caused by traffic accidents, 19 by exercise, and 7 by the crush of heavy objects. MRI of all patients showed LFN depth was more than 1.5 mm after injury, and no intervention for LFN was performed during surgery. Preoperative and postoperative depth, area, and volume of LFN defects were observed by MRI data. International Cartilage Repair Society (ICRS) score, Lysholm score, Tegner activity levels, and knee injury and osteoarthritis outcome score (KOOS) were analyzed before and after the operation.
RESULTS:
All patients were followed up from 2 to 6 years with an average of (3.28±1.12) years. There was no significant difference in the defect depth of LFN from (2.31±0.67) mm before the operation to (2.53±0.50) mm at follow-up (P=0.136). The defect area of LFN was decreased from (207.55±81.01)mm2 to (171.36±52.69)mm2 (P=0.038), and the defect volume of LFN was decreased from (426.32±176.54) mm3 to (340.86±151.54)mm3 (P=0.042). The ICRS score increased from (1.51±0.34) to (2.92±0.33) (P<0.001), the Lysholm score increased from (35.37±10.54) to (94.46±8.45) (P<0.001), and the Tegner motor score increased from (3.45±0.94) to (7.56±1.28), which was significantly higher than that of the preoperative data (P<0.001). The KOOS score of the final follow-up was 90.42±16.35.
CONCLUSION
With the increase of recovery time after anterior cruciate ligament reconstruction, the defect area and volume of LFN decreased gradually, but the defect depth remained unchanged. The knee joint function of the patients significantly improved. The cartilage of the LFN defect improved, but the repair effect was not good.
Male
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Female
;
Humans
;
Young Adult
;
Adult
;
Adolescent
;
Middle Aged
;
Anterior Cruciate Ligament Injuries/surgery*
;
Retrospective Studies
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Magnetic Resonance Imaging
;
Anterior Cruciate Ligament Reconstruction
;
Femur/surgery*
;
Treatment Outcome
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Knee Joint/surgery*
6.Effect and Mechanism of Cuscutae Semen Total Flavonoids Combined with Tripterygium wilfordii Polyglycoside Tablets on Ovarian Germline Stem Cells of Physiological Mice by Notch Signaling Pathway
Yibo YANG ; Ying XU ; Lu WANG ; Zhao ZOU ; Panyu XU ; Hanqian DU ; Jiashan LI ; Na LIN
Chinese Journal of Experimental Traditional Medical Formulae 2023;29(5):9-15
ObjectiveTo observe the effect of Cuscutae Semen total flavonoids combined with Tripterygium wilfordii polyglycoside tablets (TWPT) on ovarian germline stem cells of female physiological mice through neurogenic locus notch homolog (Notch) signaling pathway. MethodSixty female Kunming mice (5 weeks old) were randomly divided into normal group, Tripterygium wilfordii polyglycoside tablets low-, high-dose groups (13.65 mg·kg-1·d-1 and 27.3 mg·kg-1·d-1, 1 and 2 times clinical equivalent dose), Cuscutae Semen total flavonoids low- and high-dose groups (150 mg·kg-1·d-1 and 300 mg·kg-1·d-1), and combination group (13.65 mg·kg-1·d-1 TWPT and 150 mg·kg-1·d-1 Cuscutae Semen total flavonoids), with 10 in each group. After 3 weeks of continuous administration, the uterus/brain and ovarian/brain indexes were calculated, and the pathological changes of ovarian tissue were observed under light microscope. The content of estradiol in serum was determined by enzyme linked immunosorbent assay (ELISA). Immunofluorescence assay was performed to observe the expressions of germline stem cell markers in ovarian epithelium, including mouse vasa homologue (Mvh), octamer-binding transcription factor 4 (Oct4), tyrosine-protein kinase receptor (c-kit), Nanog, Notch signaling pathway molecules, neurogenic locus notch homolog protein 1 (Notch1), hes family BHLH transcription factor 1(Hes1), and jagged canonical Notch ligand 1 (JAG1). ResultCompared with the normal group, low and high doses of TWPT had no significant effect on the uterus/brain and ovary/brain indexes and the uterus and ovary morphologies of mice, while only the number of atretic follicles was increased (P<0.01). The expressions of ovarian germline stem cell markers and Notch signaling pathway molecules had a decreasing trend in TWPT low-dose group, while the expressions of Mvh, c-kit, and Nanog were down-regulated (P<0.05, P<0.01) and the expressions of Notch1 and Hes1 were also reduced (P<0.01) in TWPT high-dose group. However, the above indexes were increased in Cuscutae Semen total flavonoids low-dose group (P<0.05, P<0.01). Compared with the low does of TWPT group, the combination group had a decrease in the increased number of atretic follicles (P<0.01), an improvement in the down-regulated expressions of Mvh and Nanog (P<0.01), and an increase in the expressions of Notch1 and Hes1 (P<0.05, P<0.01). ConclusionOvarian germline stem cells are the source target of the reproductive toxicity of TWPT. Cuscutae Semen total flavonoids participate in the regulation of the germline stem cell pathways to alleviate the reproductive toxicity caused by TWPT, and its mechanism of action may be related to the Notch signaling pathway.
7.HbA1c comparison and diagnostic efficacy analysis of multi center different glycosylated hemoglobin detection systems.
Ping LI ; Ying WU ; Yan XIE ; Feng CHEN ; Shao qiang CHEN ; Yun Hao LI ; Qing Qing LU ; Jing LI ; Yong Wei LI ; Dong Xu PEI ; Ya Jun CHEN ; Hui CHEN ; Yan LI ; Wei WANG ; Hai WANG ; He Tao YU ; Zhu BA ; De CHENG ; Le Ping NING ; Chang Liang LUO ; Xiao Song QIN ; Jin ZHANG ; Ning WU ; Hui Jun XIE ; Jina Hua PAN ; Jian SHUI ; Jian WANG ; Jun Ping YANG ; Xing Hui LIU ; Feng Xia XU ; Lei YANG ; Li Yi HU ; Qun ZHANG ; Biao LI ; Qing Lin LIU ; Man ZHANG ; Shou Jun SHEN ; Min Min JIANG ; Yong WU ; Jin Wei HU ; Shuang Quan LIU ; Da Yong GU ; Xiao Bing XIE
Chinese Journal of Preventive Medicine 2023;57(7):1047-1058
Objective: Compare and analyze the results of the domestic Lanyi AH600 glycated hemoglobin analyzer and other different detection systems to understand the comparability of the detection results of different detectors, and establish the best cut point of Lanyi AH600 determination of haemoglobin A1c (HbA1c) in the diagnosis of diabetes. Methods: Multi center cohort study was adopted. The clinical laboratory departments of 18 medical institutions independently collected test samples from their respective hospitals from March to April 2022, and independently completed comparative analysis of the evaluated instrument (Lanyi AH600) and the reference instrument HbA1c. The reference instruments include four different brands of glycosylated hemoglobin meters, including Arkray, Bio-Rad, DOSOH, and Huizhong. Scatter plot was used to calculate the correlation between the results of different detection systems, and the regression equation was calculated. The consistency analysis between the results of different detection systems was evaluated by Bland Altman method. Consistency judgment principles: (1) When the 95% limits of agreement (95% LoA) of the measurement difference was within 0.4% HbA1c and the measurement score was≥80 points, the comparison consistency was good; (2) When the measurement difference of 95% LoA exceeded 0.4% HbA1c, and the measurement score was≥80 points, the comparison consistency was relatively good; (3) The measurement score was less than 80 points, the comparison consistency was poor. The difference between the results of different detection systems was tested by paired sample T test or Wilcoxon paired sign rank sum test; The best cut-off point of diabetes was analyzed by receiver operating characteristic curve (ROC). Results: The correlation coefficient R2 of results between Lanyi AH600 and the reference instrument in 16 hospitals is≥0.99; The Bland Altman consistency analysis showed that the difference of 95% LoA in Nanjing Maternity and Child Health Care Hospital in Jiangsu Province (reference instrument: Arkray HA8180) was -0.486%-0.325%, and the measurement score was 94.6 points (473/500); The difference of 95% LoA in the Tibetan Traditional Medical Hospital of TAR (reference instrument: Bio-Rad Variant II) was -0.727%-0.612%, and the measurement score was 89.8 points; The difference of 95% LoA in the People's Hospital of Chongqing Liang Jiang New Area (reference instrument: Huizhong MQ-2000PT) was -0.231%-0.461%, and the measurement score was 96.6 points; The difference of 95% LoA in the Taihe Hospital of traditional Chinese Medicine in Anhui Province (reference instrument: Huizhong MQ-2000PT) was -0.469%-0.479%, and the measurement score was 91.9 points. The other 14 hospitals, Lanyi AH600, were compared with 4 reference instrument brands, the difference of 95% LoA was less than 0.4% HbA1c, and the scores were all greater than 95 points. The results of paired sample T test or Wilcoxon paired sign rank sum test showed that there was no statistically significant difference between Lanyi AH600 and the reference instrument Arkray HA8180 (Z=1.665,P=0.096), with no statistical difference. The mean difference between the measured values of the two instruments was 0.004%. The comparison data of Lanyi AH600 and the reference instrument of all other institutions had significant differences (all P<0.001), however, it was necessary to consider whether it was within the clinical acceptable range in combination with the results of the Bland-Altman consistency analysis. The ROC curve of HbA1c detected by Lanyi AH600 in 985 patients with diabetes and 3 423 patients with non-diabetes was analyzed, the area under curve (AUC) was 0.877, the standard error was 0.007, and the 95% confidence interval 95%CI was (0.864, 0.891), which was statistically significant (P<0.001). The maximum value of Youden index was 0.634, and the corresponding HbA1c cut point was 6.235%. The sensitivity and specificity of diabetes diagnosis were 76.2% and 87.2%, respectively. Conclusion: Among the hospitals and instruments currently included in this study, among these four hospitals included Nanjing Maternity and Child Health Care Hospital in Jiangsu Province (reference instrument: Arkray HA8180), Tibetan Traditional Medical Hospital of TAR (reference instrument: Bio-Rad Variant Ⅱ), the People's Hospital of Chongqing Liang Jiang New Area (reference instrument: Huizhong MQ-2000PT), and the Taihe Hospital of traditional Chinese Medicine in Anhui Province (reference instrument: Huizhong MQ-2000PT), the comparison between Lanyi AH600 and the reference instruments showed relatively good consistency, while the other 14 hospitals involved four different brands of reference instruments: Arkray, Bio-Rad, DOSOH, and Huizhong, Lanyi AH600 had good consistency with its comparison. The best cut point of the domestic Lanyi AH600 for detecting HbA1c in the diagnosis of diabetes is 6.235%.
Pregnancy
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Child
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Humans
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Female
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Glycated Hemoglobin
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Cohort Studies
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Diabetes Mellitus/diagnosis*
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Sensitivity and Specificity
;
ROC Curve
8.Analysis of Pharmacodynamic Material Basis of Qingyan Formula Based on UPLC-Q-Orbitrap-MS and Molecular Docking
Zhao ZOU ; Jiashan LI ; Ying XU ; Yibo YANG ; Panyu XU ; Hanqian DU ; Weifeng ZHU ; Na LIN
Chinese Journal of Experimental Traditional Medical Formulae 2022;28(22):159-166
ObjectiveTo study the constituents migrating to blood of Qingyan formula by serum pharmacochemistry, and investigate the binding energy between these constituents and estrogen receptor (ER), so as to confirm the pharmacodynamic material basis of Qingyan formula in rats. MethodUltra-high performance liquid chromatography-quadrupole electrostatic field-orbital trap high resolution mass spectrometry (UPLC-Q-Orbitrap-MS) was used to determine the constituents migrating to blood of Qingyan formula in rats by comparing the fingerprint differences of 70% ethanol extract of Qingyan formula, 70% ethanol extract of each single drug in this formula, blank serum and serum after administration of 70% ethanol extract of Qingyan formula, according to the retention time, relative molecular weight and the primary and secondary ion fragments provided by MS. Mobile phase was 0.1% formic acid aqueous solution (A)-acetonitrile(B) for gradient elution (0-5 min, 2%-20%B; 5-10 min; 20%-50%B; 10-15 min, 50%-80%B; 15-25 min, 80%-95%B; 25-26 min, 95%-2%B; 26-30 min, 2%B), the flow rate was 0.3 mL·min-1 and the injection volume was 5 μL, electrospray ionization was used with detection range of m/z 150-2 000, positive and negative ion scanning modes. Molecular docking technology was used to characterize the binding energy of constituents migrating to blood with ERα and ERβ, and to confirm the material basis of this formula. ResultAfter oral administration of Qingyan formula, 30 components were detected in serum, of which 9 were prototype components and 21 were metabolites. Nine prototype components were identified as monotropein, asperuloside, verbascoside, β-ecdysone, allantoin, deacetyl asperuloside acid, echinacoside, betaine and caffeic acid, 21 metabolites mainly included organic acids, amino acids, cholines and so on. The binding energies of the above 9 prototype components with ERα were -6.7, -8.9, -6.0, -5.7, -5.3, -4.9, -7.3, -3.3, -6.3 kcal·mol-1 (1 kcal≈4 184 J), and the binding energies of them with ERβ were -6.6, -7.2, -7.7, 8.0, -7.4, -5.5, -6.9, -3.6, -6.4 kcal·mol-1, respectively. ConclusionThese nine prototype components into blood are the active ingredients of Qingyan formula that play estrogen-like role in the body, which can provide experimental basis for the formulation of quality standards and subsequent research and development of Qingyan formula.
9.Role of karyotyping combined with chromosomal microarray analysis in prenatal diagnosis of balanced translocation or inversion carriers
Wei ZHAO ; Siying LIANG ; Ning XIE ; Nan JIANG ; Yan MIAO ; Jiashan LI ; Shuo LI
Chinese Journal of Perinatal Medicine 2022;25(1):35-41
Objective:To explore the value of karyotyping and chromosomal microarray analysis (CMA) in the prenatal diagnosis of balanced translocation/inversion carriers.Methods:This was a retrospective study involving 117 balanced translocation/inversion carrier couples. Among them, 90 women had a history of spontaneous abortion(≥2 times), stillbirth, fetal multiple malformations, or giving birth to children with chromosome abnormality disease and the peripheral blood karyotyping and fluorescence in situ hybridization testing confirmed that one partner was balanced translocation/inversion carrier. The present pregnancies of these cases were spontaneous and lasted until 18-25 weeks. The other 27 cases were confirmed by chromosome examination at the present pregnancy after the indication of fetal structural abnormalities by fetal karyotyping due to advanced maternal age and abnormal ultrasound and prenatal screening results. The results of karyotyping and CMA by amniocentesis during 18 to 25 gestational weeks were all summarized and described. Results:The successful rate of both methods was 100.0% (117/117). Unbalanced and balanced translocation/inversion were detected in seven (6.0%) and 39 (33.3%) fetuses by karyotyping, respectively. CMA revealed 14 fetuses with pathogenic copy number variation (CNV) and one with variants of uncertain significance(VUS), with an anomaly detection rate of 12.8% (15/117). Among the 15 cases with CNV, 13 were related to the parental translocation/inversion, one with de novo mutation (22q11.2 microdeletion syndrome), and one Duchenne muscular dystrophy mutation carrier. Based on the results of karyotype and CMA, there were 12 fetuses with unbalanced chromosomal fragments (10.3%), 37 fetuses with balanced translocation/inversion (31.6%), and 68 fetuses with normal chromosomes (58.1%). Conclusions:The combination of karyotyping and CMA can provide more accurate prenatal genetic diagnosis when one of a couple carries balanced chromosomal translocations/inversion.
10.Influence of age on advanced neoplasia detection in colorectal cancer screening in population at high risk.
Jin Hua YANG ; Xing Lin FEI ; Qi Long LI ; Zeng Hao XU ; Kai GAO ; Hao BAI ; Jia Yu LI ; Meng Ling TANG ; Jian Bin WANG ; Ming Juan JIN ; Kun CHEN
Chinese Journal of Epidemiology 2022;43(8):1282-1287
Objective: To compare the detection rate of advanced neoplasia and the number of people needing endoscopy in colorectal cancer screening giving at different starting age in population at high risk. Methods: Based on the screening project of early diagnosis and treatment of colorectal cancer in Jiashan county, Zhejiang province, two rounds of colorectal cancer screening were conducted between January 2007 and December 2020. After excluding participants who were not at high risk or had incomplete information, 27 130 participants and 31 205 participants were finally enrolled in round one and in round two, respectively. The spline analysis based on the generalized additive model was used to describe the trend of detection rate of advanced neoplasia with age. The detection rate and number of people needing endoscopy for the groups with starting age at 50, 45 and 40 years were calculated, and the differences in the detection rate were tested by χ2 goodness of fit test. Results: A total of 21 077 (77.69%) participants in round one and 25 249 (80.91%) participants in round two received endoscopy, in whom 1 097 (detection rate=52.05‰) and 1 151 (detection rate=45.59‰) had advanced neoplasia (cancers and advanced adenomas), respectively. The detection rate increased significantly with age, and the detection rate in round one were significantly higher than that in round two (P<0.05). The overall detection rates of advanced neoplasia for the groups with starting age at 50, 45 and 40 years were 61.11‰, 56.14‰ and 52.05‰ in round one, and 49.10‰, 46.75‰ and 45.59‰ in round two, respectively. The rates were significantly higher for the group with starting age at 50 years than that with starting age at 40 years in both round one and round two (P<0.05). The numbers of people needing endoscopy of advanced neoplasia for the groups with starting age at 50, 45 and 40 years were 17, 18, and 20 in round one, and 21, 22 and 22 in round two. Conclusions: The detection rate of advanced neoplasia increased with age. Starting screening at lower age might contribute to decreased detection rate and increased number of people needing endoscopy. However, the difference was limited.
Adult
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Colorectal Neoplasms/epidemiology*
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Early Detection of Cancer
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Humans
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Mass Screening
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Middle Aged
;
Occult Blood

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