1.Effect of arterial baroreflex on survival rate of rats with cecal ligation and puncture-induced sepsis
Keyong SHI ; Xiujuan MA ; Yinglin CAO ; Wei ZHANG ; Fuming SHEN
Academic Journal of Second Military Medical University 2000;0(07):-
Objective:To investigate the effect of arterial baroreflex(ABR)on survival rate of rats with cecal ligation and puncture(CLP)-induced sepsis.Methods:Male Sprague-Dawley rats were divided into 2 groups:sham-operated rats(n=22)and sinoaortic denervated(SAD)rats(n=22).Four weeks after SAD rats were subjected to CLP-induced sepsis,the blood pressure and heart period(HP)were monitored for 12 hours in conscious state and the survival of rats was observed.Results:Both the diastolic and systolic blood pressue gradually decreased after CLP;the HP shortened first and then drastically prolonged until the death of rats.At 12 h after CLP the survival rate of SAD rats was lower than that of the sham-operated rats(59% vs 86%).Significant differences were found between the Kaplan-Meier survival curves of the rats in 2 groups(P
2.Metabolism and Transport of 7,4'-dihydroxyflavone in Caco-2 Cell Model
Yinglin MA ; Yiping ZHOU ; Yu ZHOU ; Kunyu JIANG ; Shengnan MENG
Herald of Medicine 2017;36(2):127-131
Objective To study the effects of different transport protein on the transport of 7,4'-dihydroxyflavone (7,4'-DHF) and its metabolite (7,4'-DHF-S) in Caco-2 cell model.Methods Ultra performance liquid chromatography was employed to determinethe content of 7,4'-DHF and 7,4'-DHF-S incubation buffer,their structures were identified by LC-MS/MS.Bidirectional transport of Caco-2 cells model was used to investigate the influence of ko143 (the inhibitor of BCRP) and MK571 (the inhibitor of MRP2) on the transport of 7,4'-DHF and 7,4'-DHF-S,respectively.Results Metabolic product of 7,4'-DHF in Caco-2 monolayer cell was identified as one monosulfate;PDR of 7,4'-DHF was (1.43 ± 0.11),PDR of ko143 and MK571 on the apparent permeability of 7,4'-DHF was (1.59 ± 0.04) and (1.48 ± 0.07) (P > 0.05);PDR of 7,4'-DHF-S was (1.60 ± 0.06);ko143 could significantly reduce the apparent permeability of 7,4'-DHF-S,and the PDR was (0.23 ±0.03) (P < 0.01);MK571 had no significant effect on the apparent permeability of the 7,4'-DHF-S,and the PDR was (1.51±0.04) (P > 0.05).Conclusion Caco-2 cells can mediate the suffonated reaction of 7,4'-DHF;7,4'-dihydroxyflavone sulfonated combination product may be a substrate for BCRP.
3.Characteristics and Classification of Cerebral Glucose Metabolic Decreases in Adults with Hypoxic-ischemic Encephalopathy
Yusheng SU ; Yunchuan MA ; Jianwen SHANG ; Man WANG ; Yinglin ZHANG
Chinese Journal of Rehabilitation Theory and Practice 2012;18(8):721-723
Objective To study the characteristics of cerebral glucose metabolism in adults with hypoxic-ischemic encephalopathy (HIE) using positron emission tomography (PET) and statistical parameter mapping (SPM), and evaluate the degree of the cerebral glucose metabolism damaged. Methods 26 HIE patients and 20 healthy controls received 18F-FDG PET imaging. The scope and degree of the brain radioactivity decrease were observed with visual analysis. The three-dimensional projection images and the KE value were obtained by SPM analysis. Results The glucose metabolic decrease in HIE was primarily bilaterally. The bilateral basal ganglia and thalamus metabolism decreased most obviously. The brain cortical lobes varied degrees of metabolic decrease according to the order from high to low was the frontal, occipital, parietal and temporal lobes. The basal ganglia and thalamus were taken as the important target area of the evaluation of damage degree, the degree of damage of HIE was divided into Level Ⅰ(mild), Level Ⅱ(moderate) and Level Ⅲ (severe) combined with cortex damage. Conclusion The basal ganglia and thalamus are the target areas of metabolic damage, the classification combined with brain cortex damage degree and scope can be used to guide the clinical treatment and prognosis evaluation.
4.Study on kinectics characteristics of the sulfation of apigenin by SULTIA3
Kunyu JIANG ; Xiaoyue LV ; Yu ZHOU ; Yiping ZHOU ; Yinglin MA ; Shengnan MENG
Chinese Journal of Biochemical Pharmaceutics 2015;(3):153-155,158
Objective To investigate the kinectics characteristics of sulfation of apigenin mediated by SULTIA3.Methods After incubation of apigenin using in vitro SULT1A3 system, high-performance liquid chromatography was utilized to determine the sulfates of apigenin.Mass spectrum(MS) were employed to elucidate the structure of metabolite.The program GraphPad Prism 5 was used to perform the kinetic characterization of SULT1A3 catalyzed metabolism of apigenin.Results A liner calibration curve for the assay of apigenin was validated in the range of 0.15625 ~30 μM with the recoveries of at least 80% and intra-day and inter-day RSD of less than 15%.Metabolic product of apigenin and SULT1A3 in the incubated system was identified one monosulfate.The metabolic behavior of apigenin in SULT1A3 was followed substrate inhibition kinetics.Apparent kinetic parameters of metabolism of apigenin by SULT1A3, Kmwas(0.355 ±1.04) μM and Ksi was(23.62 ±0.06) μM,Vmax was(65.71 ±1.30) nmol/(min? mg),Vmax/Km was 185.10 mL/(min? mg).Conclusion SULT1A3 can mediate the binding of apigenin sulfonated reaction, and the character of enzymatic kinetics shows substrate inhibition.Sulfation of apigenin mediated by SULTIA3 may play an important role in phaseⅡmetabolic in vivo.
5.Metabolism profile of sulfation of chrysin in human small intestine S9
Kunyu JIANG ; Yiping ZHOU ; Yinglin MA ; Yu ZHOU ; Maofan ZHANG ; Shengnan MENG
Chinese Journal of Biochemical Pharmaceutics 2015;37(4):154-157
Objective To investigate the metabolic profile of chrysin in SULT1A3 and human small intestine S9.Methods After incubation of chrysin using in vitro SULT1A3 and human small intestine S9 system, high-performance liquid chromatography was utilized to determine the sulfates of chrysin.Mass spectrum(MS) were employed to elucidate the structures of metabolite.Results In the SULT1A3 with PAPS, Km were (3.06 ±1.04) and (0.41±0.06) μM, Vmax were (12.13 ±1.30) and (6.72 ±1.61) nmol/(min· mg), Vmax/Km were 3.96 and 16.39 mL/(min· mg), respectively.In the human small intestine S9 with PAPS, Km were (1.92 ±0.35) and (0.01 ±0.00) μM, Vmax were (0.52 ±0.02) and (0.08 ± 0.02) nmol/(min· mg), Vmax/Km were 0.27 and 8.00 mL/(min· mg).The metabolic behavior of chrysin in SULT1A3 and human small intestine S9 both were followed biphasic kinetics.The sulfation of chrysin in SULT1A3 showed a significant correlation with that in human small intestine S9(R2 =0.985).Conclusion The result indicates that SULT1A3 is the major enzyme to the metabolism of chrysin, human small intestine may be the main metabolic organs of chrysin.
6.Effect of TNF-related apoptosis inducing ligand on the biological activity of hepatocarcinoma cell line
Lihui HAN ; Wensheng SUN ; Suxia LIU ; Xiaoqing JIA ; Xiaoyan WANG ; Chunhong MA ; Lifen GAO ; Lining ZHANG ; Yinglin CAO
Chinese Journal of Pathophysiology 2000;0(08):-
AIM: To explore the effect of TNF-related apoptosis inducing ligand (TRAIL), a new apoptotic inducing molecule on the biological activity of hepatocarcinoma cell line. METHODS: The expression of membrane binding TRAIL on HepG2 cells was detected by immuno-cytochemistry. Quantity of secretory TRAIL was assayed by ELISA method. The cytotoxicity and apoptosis induced by TRAIL was detected by MTT and TUNEL method, respectively. The telomerase activity of HepG2 cells was detected by TRAP-PCR assay kit. The expression of hTERT, the catalytic subunit of telomerase, was detected by FCM. RESULTS: TRAIL was constitutively expressed on the membrane of HepG2 cell line. Soluble TRAIL was also expressed to a certain degree. Cytotoxicity assay showed that TRAIL significantly inhibited the growth of hepatocarcinoma cells. TUNEL assay indicated that TRAIL induced apoptosis in hepatocarcinoma cells. Detection of telomerase activity showed that TRAIL inhibited telomerase activity and the expression of telomerase catalytic subunit. CONCLUSION: TRAIL is an effective molecule to inhibit the growth of hepatocarcinoma through multiple pathways, such as inducing apoptosis and inhibiting the activity of telomerase.
7.Effect of preS2 antisense RNA on hepatocellular carcinoma with a novel delivery system.
Chunhong MA ; Wensheng SUN ; Peikun TIAN ; Xiaoyan WANG ; Suxia LIU ; Lining ZHANG ; Yinglin CAO ; Faliang ZHU ; Qiu ZHANG
Chinese Medical Journal 2003;116(5):717-720
OBJECTIVESTo construct a hepatoma directed gene delivery system which could transfer preS2 antisense RNA to liver cancer cells specifically, and to explore a new therapeutic strategy for hepatocellular carcinoma by blocking hepatitis B virus (HBV) with antisense RNA targeting hepatocellular carcinoma.
METHODSGE7 and HA20 were synthesized and mixed with pEBAF-as-preS2, a hepatocarcinoma specific HBV antisense expression vector, to construct a novel HBV antisense RNA delivery system named AFP-enhancing 4-element complex. Nude mice bearing hepatocelluar carcinoma cells HepG2.2.15 were injected with AFP-enhancing 4-element complex via a tail vein. Total RNA from tissues was extracted, and reversal transcription-ploymerase chain reaction (RT-PCR) was used to detect the expression of preS2. Different doses of AFP-enhancing 4-element complex was injected into nude mice at different time points, and tumor diameter was measured.
RESULTSAFP-enhancing 4-element complex was constructed successfully. RT-PCR showed preS2 antisense RNA delivered by AFP-enhancing 4-element complex only expressed in liver tumor HepG2.2.15 cells of the mice. After the treatment of AFP-enhancing 4-element complex with dose of 0.2 micro g per mouse (once a week for 4 weeks), the mean tumor diameter of nude mice was significantly shorter than that of the control groups (0.995 +/- 0.35 cm vs 2.125 +/- 0.25 cm, P < 0.01).
CONCLUSIONSAn HBV antisense RNA gene delivery system targeting hepatocellular carcinoma, AFP-enhancing 4-element complex, was constructed successfully. PreS2 antisense RNA expressed specifically in hepatocelluar carcinoma cells significantly inhibits tumor growth of mice bearing hepatocarcinoma HepG2.2.15 and may have therapeutic potential in HBV related hepatocarcinoma.
Animals ; Drug Delivery Systems ; Gene Transfer Techniques ; Genetic Therapy ; Genetic Vectors ; Hepatitis B Surface Antigens ; therapeutic use ; Hepatitis B virus ; genetics ; Liver Neoplasms, Experimental ; pathology ; therapy ; Male ; Mice ; Mice, Nude ; Protein Precursors ; therapeutic use ; RNA, Antisense ; therapeutic use
8.Study on the Mechanism of Yiqi Tongmai Powder Against Cerebral Ischemia-Reperfusion Injury Based on Network Pharmacology,Molecular Docking and Experimental Verification
Pengpeng SONG ; Yanke GUO ; Dongsheng GUAN ; Ming MA ; Yinglin CUI
Traditional Chinese Drug Research & Clinical Pharmacology 2024;35(7):1016-1027
Objective To investigate the mechanism of Yiqi Tongmai Powder(Ginseng Radix et Rhizoma,Salviae Miltiorrhizae Radix et Rhizoma,Notoginseng Radix et Rhizoma,Hirudo,Eupolyphaga Steleophaga,Rhei Radix et Rhizoma)against cerebral ischemia-reperfusion injury(CIRI)based on network pharmacology,molecular docking and experimental verification.Methods(1)The active components of Yiqi Tongmai Powder and their action targets were screened by TCMSP,TCMID and ETCM databases,the disease related targets of CIRI were screened by GeneCards,OMIM and TTD disease databases,and the intersection targets of the above targets were obtained through Venny 2.1 online platform,that is,the potential targets of Yiqi Tongmai Powder in the treatment of CIRI.The"drugs-active components-targets"network was constructed by Cytoscape 3.7.1 software,and the potential active components of Yiqi Tongmai Powder in the treatment of CIRI were screened.Protein-protein interaction(PPI)analysis of potential targets was carried out by STRING 11.0 database to screen core targets.The GO function and KEGG pathway enrichment of potential targets were analyzed by Metascape database.AutoDockTools 1.5.7 software was used to verify the molecular docking between the key active components and the core targets.(2)The rat model of CIRI was established by middle cerebral artery occlusion and reperfusion(MCAO/R).SD rats were randomly divided into sham operation group,model group,Yiqi Tongmai Powder low-dose group(0.27 g·kg-1),Yiqi Tongmai Powder high-dose group(1.08 g·kg-1)and Nimodipine group(30 mg·kg-1),with 10 rats in each group.Pre-administration began three days before the establishment of the model,once a day for seven days.The neurological deficit of MCAO/R rats was evaluated by modified neurological deficit score(mNSS).The volume of cerebral infarction was measured by TTC staining.Nissl staining was used to observe the damage of neurons in brain tissue.ELISA method was used to detect serum inflammatory factors and oxidative stress related indexes.TUNEL staining was used to detect brain tissue apoptosis.Western Blot method was used to detect the protein expressions of Bax and Bcl-2 in brain tissue.Results(1)A total of 46 active components and 178 potential targets of Yiqi Tongmai Powder in the treatment of CIRI were obtained.The key active components such as quercetin,digitalis flavonoids,kaempferol,valine and uracil were analyzed,and the core targets such as TNF,IL-6,STAT3,VEGFA,AKT1,IL-1β,CASP3,TP53,MAPK3 and EGFR were analyzed.The potential targets are involved in inflammation,oxidative stress,cell proliferation and differentiation,apoptosis and other biological processes,including cAMP,NF-κB,PI3K-Akt and other signal pathways.The main active components quercetin,flavonoids of digitalis,kaempferol and valine have good binding activity to target proteins such as TNF,IL-6,STAT3 and VEGFA.(2)Compared with the model group,the neurological deficit score of rats in each treatment group was significantly decreased(P<0.05,P<0.01),the area of cerebral infarction was significantly decreased(P<0.01),and the pathological changes of ischemic necrotic area of brain tissue were improved.The number of neurons in ischemic area of brain tissue was significantly increased(P<0.01),and the rate of neuronal apoptosis was significantly decreased(P<0.01).The levels of serum IL-1β,IL-6,TNF-α and MDA were significantly decreased,while the activities of SOD and GSH-Px were significantly increased(P<0.05).The protein expression of Bax in brain tissue were significantly decreased and the protein expression of Bcl-2 significantly increased(P<0.05).Conclusion Yiqi Tongmai Powder may play an anti-CIRI effect by reducing inflammation and oxidative stress,inhibiting cell apoptosis.
9.Multicenter evaluation of the diagnostic efficacy of jaundice color card for neonatal hyperbilirubinemia
Guochang XUE ; Huali ZHANG ; Xuexing DING ; Fu XIONG ; Yanhong LIU ; Hui PENG ; Changlin WANG ; Yi ZHAO ; Huili YAN ; Mingxing REN ; Chaoying MA ; Hanming LU ; Yanli LI ; Ruifeng MENG ; Lingjun XIE ; Na CHEN ; Xiufang CHENG ; Jiaojiao WANG ; Xiaohong XIN ; Ruifen WANG ; Qi JIANG ; Yong ZHANG ; Guijuan LIANG ; Yuanzheng LI ; Jianing KANG ; Huimin ZHANG ; Yinying ZHANG ; Yuan YUAN ; Yawen LI ; Yinglin SU ; Junping LIU ; Shengjie DUAN ; Qingsheng LIU ; Jing WEI
Chinese Journal of Pediatrics 2024;62(6):535-541
Objective:To evaluate the diagnostic efficacy and practicality of the Jaundice color card (JCard) as a screening tool for neonatal jaundice.Methods:Following the standards for reporting of diagnostic accuracy studies (STARD) statement, a multicenter prospective study was conducted in 9 hospitals in China from October 2019 to September 2021. A total of 845 newborns who were admitted to the hospital or outpatient department for liver function testing due to their own diseases. The inclusion criteria were a gestational age of ≥35 weeks, a birth weight of ≥2 000 g, and an age of ≤28 days. The neonate′s parents used the JCard to measure jaundice at the neonate′s cheek. Within 2 hours of the JCard measurement, transcutaneous bilirubin (TcB) was measured with a JH20-1B device and total serum bilirubin (TSB) was detected. The Pearson′s correlation analysis, Bland-Altman plots and the receiver operating characteristic (ROC) curve were used for statistic analysis.Results:Out of the 854 newborns, 445 were male and 409 were female; 46 were born at 35-36 weeks of gestational age and 808 were born at ≥37 weeks of gestational age. Additionally, 432 cases were aged 0-3 days, 236 cases were aged 4-7 days, and 186 cases were aged 8-28 days. The TSB level was (227.4±89.6) μmol/L, with a range of 23.7-717.0 μmol/L. The JCard level was (221.4±77.0) μmol/L and the TcB level was (252.5±76.0) μmol/L. Both the JCard and TcB values showed good correlation ( r=0.77 and 0.80, respectively) and agreements (96.0% (820/854) and 95.2% (813/854) of samples fell within the 95% limits of agreement, respectively) with TSB. The JCard value of 12 had a sensitivity of 0.93 and specificity of 0.75 for identifying a TSB ≥205.2?μmol/L, and a sensitivity of 1.00 and specificity of 0.35 for identifying a TSB ≥342.0?μmol/L. The TcB value of 205.2?μmol/L had a sensitivity of 0.97 and specificity of 0.60 for identifying TSB levels of 205.2 μmol/L, and a sensitivity of 1.00 and specificity of 0.26 for identifying TSB levels of 342.0 μmol/L. The areas under the ROC curve (AUC) of JCard for identifying TSB levels of 153.9, 205.2, 256.5, and 342.0 μmol/L were 0.96, 0.92, 0.83, and 0.83, respectively. The AUC of TcB were 0.94, 0.91, 0.86, and 0.87, respectively. There were both no significant differences between the AUC of JCard and TcB in identifying TSB levels of 153.9 and 205.2 μmol/L (both P>0.05). However, the AUC of JCard were both lower than those of TcB in identifying TSB levels of 256.5 and 342.0 μmol/L (both P<0.05). Conclusions:JCard can be used to classify different levels of bilirubin, but its diagnostic efficacy decreases with increasing bilirubin levels. When TSB level are ≤205.2 μmol/L, its diagnostic efficacy is equivalent to that of the JH20-1B. To prevent the misdiagnosis of severe jaundice, it is recommended that parents use a low JCard score, such as 12, to identify severe hyperbilirubinemia (TSB ≥342.0 μmol/L).