1.A reporter gene assays for bioactivity determination of human chorinonic gonadotropin
Ying HUANG ; Xiao-ming ZHANG ; He-yang LI ; Lü-yin WANG ; Hui ZHANG ; Ping LÜ ; Jing LI ; Xiang-dong GAO ; Cheng-gang LIANG
Acta Pharmaceutica Sinica 2024;59(2):432-438
This study constructed a LHCGR-CRE-luc-HEK293 transgenic cell line according to the activation of the cAMP signaling pathway after recombinant human chorionic gonadotropin binding to the receptor. The biological activity of recombinant human chorionic gonadotropin was assayed using a luciferase assay system. The relative potency of the samples was calculated using four-parameter model. And the method conditions were optimized to validate the specificity, relative accuracy, precision and linearity of the method. The results showed that there was a quantitative potency relationship of human chorinonic gonadotropin (hCG) in the method and it was in accordance with the four-parameter curve. After optimization, the conditions were determined as hCG dilution concentration of 2.5 μg·mL-1, dilution ratio of 1∶4, cell number of 10 000-15 000 cells/well, and induction time of 6 h. The method had good specificity, relative accuracy with relative bias ranging from -8.9% to 3.4%, linear regression equation correlation coefficient of 0.996, intermediate precision geometric coefficient of variation ranging from 3.3% to 15.0%, and linearity range of 50% to 200%. This study successfully established and validated a reporter gene method to detect hCG biological activity, which can be used for hCG biological activity assay and quality control.
2.Data Mining of Medication Rules for the Treatment of Atopic Dermatitis the Children by Chinese Medical Master XUAN Guo-Wei
Jin-Dian DONG ; Cheng-Cheng GE ; Yue PEI ; Shu-Qing XIONG ; Jia-Fen LIANG ; Qin LIU ; Xiu-Mei MO ; Hong-Yi LI
Journal of Guangzhou University of Traditional Chinese Medicine 2024;41(3):752-758
Objective Data mining technology was used to mine the medication rules of the prescriptions used in the treatment of pediatric atopic dermatitis by Chinese medical master XUAN Guo-Wei.Methods The medical records of effective cases of pediatric atopic dermatitis treated by Professor XUAN Guo-Wei at outpatient clinic were collected,and then the medical data were statistically analyzed using frequency statistics,association rule analysis and cluster analysis.Results A total of 242 prescriptions were included,involving 101 Chinese medicinals.There were 23 commonly-used herbs,and the 16 high-frequency herbs(frequency>100 times)were Glycyrrhizae Radix et Rhizoma,Saposhnikoviae Radix,Glehniae Radix,Perillae Folium,Ophiopogonis Radix,Cynanchi Paniculati Radix et Rhizoma,Microctis Folium,Dictamni Cortex,Scrophulariae Radix,Coicis Semen,Cicadae Periostracum,Lilii Bulbus,Rehmanniae Radix,Kochiae Fructus,Sclerotium Poriae Pararadicis,and Euryales Semen.The analysis of the medicinal properties showed that most of the herbs were sweet and cold,and mainly had the meridian tropism of the spleen,stomach and liver meridians.The association rule analysis yielded 24 commonly-used drug combinations and 20 association rules.Cluster analysis yielded 2 core drug combinations.Conclusion For the treatment of pediatric atopic dermatitis,Professor XUAN Guo-Wei focuses on the clearing,supplementing and harmonizing therapies,and the medication principle of"supporting the healthy-qi to eliminate the pathogen,and balancing the yin and yang"is applied throughout the treatment.
3.Research advances in exosomal proteins,mRNA and non-coding RNA regulation of Hepatocellular Carci-noma
Yuxin CHENG ; Liang LIU ; Shiyu DONG ; Shengchao LI ; Meng ZHANG
The Journal of Practical Medicine 2024;40(6):748-755
Exosomes are extracellular vesicles commonly detected in numerous body fluids and contain a variety of components such as proteins,nucleic acids,lipids,and metabolites.These components enable exosomes to mediate intercellular communication and impact diverse cellular processes.Recently,research has highlighted that exosomes have a significant regulatory role in numerous aspects of hepatocellular carcinoma(HCC)occur-rence,development,and drug resistance.Non-coding RANs,a crucial component of exosomes,can regulate the HCC tumour microenvironment with a direct impact on biological behaviours such as tumour growth,metastasis,angiogenesis,and immunomodulation.To this end,exosomes present an interesting avenue for further research in the field of HCC therapy.It is anticipated to become a novel diagnostic,prognostic marker,or therapeutic target for hepatocellular carcinoma(HCC).Our review of the role played by exosomal components in HCC progression over the last five years aims to furnish references and innovative perspectives for early diagnosis,prognosis,and treatment of HCC.
4.Bioequivalence study of compound lidocaine cream in healthy Chinese subjects
Meng-Qi CHANG ; Yu-Qi SUN ; Qiu-Jin XU ; Xi-Xi QIAN ; Ying-Chun ZHAO ; Yan CAO ; Liu WANG ; Cheng ZHANG ; Dong-Liang YU
The Chinese Journal of Clinical Pharmacology 2024;40(9):1321-1326
Objective To study the pharmacokinetic characteristics of the test formulation of compound lidocaine cream and reference formulation of lidocaine and prilocaine cream in Chinese healthy subjects and to evaluate whether there is bioequivalence between the two formulations.Methods A single-center,single-dose,randomized,open-label,two-period,two-sequence,crossover design was used.This study included 40 healthy subjects,and in each period,test formulation or reference formulation 60 g was applied to the skin in front of both thighs(200 cm2 each side,a total of 400 cm2)under fasting conditions,and the drug was left on for at least 5 h after application.The concentrations of lidocaine and prilocaine in plasma were determined using liquid chromatography-tandem mass spectrometry(LC-MS/MS)method.Pharmacokinetic parameters were calculated using WinNonlin 8.0 software to evaluate the bioequivalence of the two formulations.Results After the application of the test formulation compound lidocaine cream and the reference formulation lidocaine and prilocaine cream on both thighs of the subjects,the pharmacokinetic parameters of lidocaine in plasma were as follows:Cmax were(167.27±91.33)and(156.13±66.86)ng·mL-1,AUC0-t were(1 651.78±685.09)and(1 636.69±617.23)ng·mL-1·h,AUC0-∞ were(1 669.85±684.65)and(1 654.37±618.30)ng·mL-1·h,the adjusted geometric mean ratios were 104.49%,101.88%and 101.89%,respectively,with 90%confidence intervals of 98.18%-111.20%,97.80%-106.13%and 97.87%-106.07%,all within the range of 80.00%-125.00%.The pharmacokinetic parameters of prilocaine in plasma were as follows:Cmax were(95.66±48.84)and(87.52±39.16)ng·mL-1,AUC0-t were(790.86±263.99)and(774.14±256.42)ng·mL-1·h,AUC0_m were(807.27±264.67)and(792.84±254.06)ng·mL-1 h,the adjusted geometric mean ratios were 107.34%,103.55%and 102.98%,respectively with 90%confidence intervals of 101.69%-113.31%,99.94%-107.30%and 99.65%-106.43%,all within the range of 80.00%-125.00%.Conclusion The test formulation compound lidocaine cream and the reference formulation lidocaine and prilocaine cream are bioequivalent.
5.Variation rules of main secondary metabolites in Hedysari Radix before and after rubbing strip
Xu-Dong LUO ; Xin-Rong LI ; Cheng-Yi LI ; Peng QI ; Ting-Ting LIANG ; Shu-Bin LIU ; Zheng-Ze QIANG ; Jun-Gang HE ; Xu LI ; Xiao-Cheng WEI ; Xiao-Li FENG ; Ming-Wei WANG
Chinese Traditional Patent Medicine 2024;46(3):747-754
AIM To investigate the variation rules of main secondary metabolites in Hedysari Radix before and after rubbing strip.METHODS UPLC-MS/MS was adopted in the content determination of formononetin,ononin,calycosin,calycosin-7-glucoside,medicarpin,genistein,luteolin,liquiritigenin,isoliquiritigenin,vanillic acid,ferulic acid,γ-aminobutyric acid,adenosine and betaine,after which cluster analysis,principal component analysis and orthogonal partial least squares discriminant analysis were used for chemical pattern recognition to explore differential components.RESULTS After rubbing strip,formononetin,calycosin,liquiritigenin and γ-aminobutynic acid demonstrated increased contents,along with decreased contents of ononin,calycosin-7-glucoside and vanillic acid.The samples with and without rubbing strip were clustered into two types,calycosin-7-glucoside,formononetin,γ-aminobutynic acid,vanillic acid,calycosin-7-glucoside and formononetin were differential components.CONCLUSION This experiment clarifies the differences of chemical constituents in Hedysari Radix before and after rubbing strip,which can provide a reference for the research on rubbing strip mechanism of other medicinal materials.
6.Efficacies of proximal femoral nail anti-rotation internal fixation in different body positions on elderly unstable femoral intertrochanteric fractures
Ling-Yan ZHAO ; Hong-Bo ZHAO ; Dong-Hai YANG ; Hui LIANG ; Cheng-Ming CAO ; Xiao-Ning LIU
Journal of Regional Anatomy and Operative Surgery 2024;33(3):239-243
Objective To investigate the efficacies of proximal femoral nail anti-rotation(PFNA)internal fixation in traction bed supine position and non-traction bed lateral position in the treatment of elderly unstable femoral intertrochanteric fractures.Methods The clinical data of patients with unstable femoral intertrochanteric fractures treated with PFNA internal fixation in our hospital were retrospec-tively analyzed,41 patients received treatment in traction bed supine position were included in the supine position group,and 55 patients treated received treatment in non-traction bed lateral position were included in the lateral position group.The perioperative related indicators,surgical reduction,hip Harris score,and incidence of complications in the two groups were analyzed.Results The operation time and incision length of patients in the lateral position group were shorter than those in the supine position group,and the intraoperative blood loss and fluoroscopy times were less than those in the supine position group,with statistically significant differences(P<0.05).There was no significant difference in the anesthesia mode,blood transfusion or hospital stay of patients between the two groups(P>0.05).There was no significant difference in the incidence of postoperative complications of patients between the two groups(P>0.05).There was no significant difference in neck-shaft angle,tip-apex distance or hip Harris score of patients between the two groups(P>0.05).Conclusion PFNA internal fixation in traction bed supine position and non-traction bed lateral position have the same effect in the treatment of elderly unstable femoral intertrochanteric fractures,while the non-traction bed lateral position for treatment has more advantages in shortening operation time,decreasing intraoperative blood loss,and reducing radiation exposure.
7.Curative effect of percutaneous microwave ablation therapy on hepatocellular carcinoma survival: a 15-year real-world study
Yanchun LUO ; Manlin LANG ; Wenjia CAI ; Zhiyu HAN ; Fangyi LIU ; Zhigang CHENG ; Xiaoling YU ; Jianping DOU ; Xin LI ; Shuilian TAN ; Xuejuan DONG ; Ping LIANG ; Jie YU
Chinese Journal of Hepatology 2024;32(4):332-339
Objective:To evaluate the long-term efficacy of percutaneous microwave ablation (MWA) therapy for hepatocellular carcinoma.Methods:2054 cases with Barcelona Clinic Liver Cancer (BCLC) stage 0~B at the Fifth Medical Center of the Chinese People's Liberation Army General Hospital from January 2006 to September 2020 were retrospectively collected. All patients were followed up for at least 2 years. The primary endpoint of overall survival and secondary endpoints (tumor-related survival, disease-free survival, and postoperative complications) of patients treated with ultrasound-guided percutaneous MWA were analyzed. Kaplan-Meier method was used for stratified survival rate analysis. Fine-and-Gray competing risk model was used to analyze overall survival.Results:A total of 5 503 HCC nodules [mean tumor diameter (2.6±1.6) cm] underwent 3 908 MWAs between January 2006 and September 2020, with a median follow-up time of 45.6 (24.0 -79.2) months.The technical effectiveness rate of 5 375 tumor nodules was 97.5%. The overall survival rates at 5, 10, and 15-years were 61.6%, 38.8%, and 27.0%, respectively. The tumor-specific survival rates were 67.1%, 47.2%, and 37.7%, respectively. The free tumor survival rates were 25.8%, 15.7%, and 9.9%, respectively. The incidence rate of severe complications was 2.8% (108/3 908). Further analysis showed that the technical effectiveness and survival rate over the passing three time periods from January 2006-2010, 2011-2015, and 2016-September 2020 were significantly increased, with P ?0.001, especially for liver cancer 3.1~5.0 cm ( P ?0.001). Conclusion:Microwave ablation therapy is a safe and effective method for BCLC stage 0-B, with significantly enhanced technical efficacy and survival rate over time.
8.Mechanism of andrographolide alleviates lipopolysaccharide-induced fer-roptosis in renal tubular epithelial cells
Yi-Xin ZHANG ; Ming HUANG ; Guo-Dong CAO ; You-Cheng ZENG ; Liang LIN ; Xiao-Yue WANG ; Qing-Hong CHENG
Chinese Journal of Infection Control 2024;23(5):568-573
Objective To investigate the effect and mechanism of andrographolide(AG)on lipopolysaccharide(LPS)-induced ferroptosis in renal tubular epithelial cells(HK-2 cells).Methods HK-2 cells were treated with LPS to simulate the in vitro HK-2 injury model of sepsis.The cells were further treated with AG of 5,10,20,40 μmol/L and randomly divided into control group,LPS group,LPS+dimethyl sulfoxide group(DMSO group),and AG group.Cell viability was detected by the CCK-8 method,and the optimal concentrations of LPS and AG were screened.Cell morphological change,the levels of kidney injury markers,including neutrophil gelatinase-associated lipocalin(NGAL),kidney injury molecule-1(KIM-1),malondialdehyde(MDA),glutathione(GSH)and reactive oxygen species(ROS),as well as the expression levels of ferroptosis regulatory proteins such as solute carrier family 7 member 11(SLC7A11),glutathione peroxidase 4(GPX4)and ferritin in each group were compared,and the pro-tective effect of AG treatment on the cells was evaluated.Results Compared with the control group,the cell viabi-lity and GSH content decreased significantly in HK-2 cells treated with 10 μg/mL LPS;cell shrinkage and adhesion ability were poor;the contents of oxidative products MDA and ROS,as well as the levels of kidney injury markers NGAL and KIM-1 increased significantly,while expression levels of SLC7A11 and GPX4 protein decreased;ferritin expression level increased;differences were all statistically significant(all P<0.05).Compared with LPS group,the cell viability,GSH content,as well as protein expression levels of SLC7A11 and GPX4 increased significantly after AG intervention,while ferritin expression level decreased,differences were all significant(all P<0.05).MDA content,ROS fluorescence intensity,and the levels of kidney injury markers NGAL and KIM-1 decreased sig-nificantly,difference were all significant(all P<0.05).Conclusion AG has a protective effect on LPS-induced HK-2 cell injury,possibly by activating SLC7A11/GPX4 pathway,reducing oxidative stress,up-regulating antioxi-dant enzyme activity,and alleviating ferroptosis.
9.Effect of injection volume on clinical efficacy after minimally invasive puncture and drainage in the treatment of moderate basal ganglia cerebral hemorrhage underlying controlling the dosage of drugs in the elderly
Liang WANG ; Wei DONG ; Lian ZHENG ; Jinmiao ZHANG ; Ding YUAN ; Guo CHENG
Chinese Journal of Nervous and Mental Diseases 2024;50(5):268-273
Objective To investigate the effect of injection volume on clinical efficacy after minimally invasive puncture and drainage in the treatment of moderate basal ganglia cerebral hemorrhage underlying controlling the dosage of drugs in the elderly.Methods After successfully inserting the puncture needle into the center point in the largest layer long axis,the liquid hematoma was aspirated,and 40,000 units urokinase was injected into the hematoma cavity twice a day(2 mL in the small volume Group(SG),and 4 mL in the large volume group(LG),respectively).The needle were then adjusted and removed according to the hematoma clearance.The clearance rate of hematoma,intracranial pressure,edema volume,mortality,incidence of complications,neurological deficit score(NDS)and activities of daily living(ADL)were compared and analyzed.Results There were no significant differences in the first hematoma clearance rate,postoperative ICP and mortality between the two groups(P>0.05).Comparing to the SG,there was a relative higher postoperative hematoma clearance rate and lower volume of perihematoma edema in the LG(P<0.05).Meanwhile,the needle retention time,urokinase usage and cumulative complications at 28 days after operation were lower than in the SG(P<0.05).For two parameters representing long-term outcome,the NDS and ADL scores were much better in LG than in SG(P<0.05).Conclusion Large volume injection can effectively drain the hematoma,shorten the needle retention time,reduce the usage of urokinase,decrease the incidence of postoperative complications and improve the prognosis of patients,thereby providing effective and safe method in treating moderate basal ganglia cerebral hemorrhage in the elderly.
10.Clinical value of immature granulocyte percentage in pre-dicting severity of acute appendicitis in children
Xin-Li ZHANG ; Kai-Jiang LI ; Pu-Yu ZHAO ; Liang ZHAO ; Bing LIANG ; Dong-Fang LU ; Yu-Cheng SHI
Chinese Journal of Current Advances in General Surgery 2024;27(7):533-537
Objective:To investigate the clinical value immature granulocyte percentage(IG%)and other inflammatory indicators in the severity of acute appendicitis.Methods:A total of 201 pediatric patients undergoing appendicitis surgery admitted to Zhoukou Central Hospital from June 2022 to August 2023 were included.Patients with pathologically confirmed actue appendici-tis were divided two subgroups:actue simple appendicitis(ASA)group and actue complicated ap-pendicitis(ACA)group,The variables that included IG%,white blood cell(WBC)count,absolute neutrophil count(ANC),absolute lymphocyte count(ALC),neutrophil to lymphocyte ratio(NLR),pro-calcitonin(PCT),C-reactive protein(CRP),platelet to lymphocyte(PLR)and other indexes were ana-lyzed between ASA and ACA group.The logistic regression model for diagnosis of ACA was es-tablished,and the diagostic value of this model and other inflammtory indicators for ACA was evaluated by receiver operating characteristic(ROC)curve analysis.Results:The levels of IG%,WBC,ANC,ALC,NLR,PCT and PLR were higher and the level of ALC was lower in ACA group than those in ASA group(all P<0.05).Logistic regression analysis showed that IG%,NLR and CRP were three diagnostic determinants of ACA(all P<0.05).The AUC of the established logistic model and IG%,NLR,CRP were 0.868,0.821,0.691 and 0.790(all P<0.001).The logistic model was vali-dated by independent cohorts,and the AUC was 0.872,the sensitivity was 90.0%and the speci-ficity was 75.6%.Conlusions:The IG%value can early indicator for pediatric ACA,and the es-tablished logistic regression model based on biomarkers including IG%,NLR and CRP has clinical value in diagnosing ACA in children.

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