3.Research advances in animal models of neonatal hyperbilirubinemia
Li YANG ; De WU ; Jiulai TANG
Chinese Journal of Comparative Medicine 2015;(9):78-80
Therearemanywaystoestablishanimalmodelsofneonatalhyperbilirubinemia,suchasintraperitoneal or intravenous injection , genetic defect animal models , the use of chemical drugs , and injection of bilirubin into cerebellomedullary cistern , and so on . In order to study the etiology , pathogenesis , and therapy of neonatal hyperbilirubinemia through animal models , we review the literature on rodent and primate models of neonatal hyperbilirubinemia ,including establishment of models and their applications , in order to provide reference for the research of neonatal hyperbilirubinemia .
5.Hyperbilirubinemia models caused by hemolysis in newborn rhesus monkeys
Baotian WANG ; Jiulai TANG ; Li YANG ; Keyin TIAN ; De WU
Chinese Journal of Applied Clinical Pediatrics 2016;31(15):1192-1195
Objective To establish the newborn rhesus monkey model of hemolytic hyperbilirnbinemia and provide an experimental basic model for research of hyperbilirubinemia.Methods Sixteen 3-day old newborn rhesus monkeys were divided into experimental group and control group,with 8 newborn rhesus monkeys in each group.Eight newborn rhesus monkeys in experimental group were treated with intravenous injection of l0 g/L phenylhydrazine hydrochloride (50 mg/kg) to establish model of homolytic hyperbilirubinemia.The newborn rhesus monkeys in control group were treated with intravenous injection of 9 g/L saline at the same time.Twenty-four hours and 48 hours after the experimental treatment,the bilirubin in blood was detected to evaluate the models,and the clinical manifestations of newborn rhesus monkeys with hyperbilirubinemia were recorded by using monitoring equipment.The brain slices were made to evaluate the model in 1 dead monkeys of experimental group.Results The newborn rhesus monkey of experimental group showed obvious skin,sclera jaundice and hemoglobinuria.The serum total bilirubin [(252.76 ± 63.42) μmol/L],unconjugated bilirubin[(165.85 ±44.93) pmol/L] and conjugated bilirubin [(87.16 ±21.22) μmol/L] in the experimental group were significantly higher than those [(20.62 ± 5.72) μmol/L,(7.93 ± 2.31) μmol/L,(12.51 ± 3.53) μmol/L] in the control group,and the differences were statistically significant (t =14.581,13.881,14.040,all P < 0.01).The level of hemoglobin [(47.18 ± 10.09) μmol/L] in the experimental group was significantly lower than that of the control group [(136.85 ± 13.48) μmol/L],and the difference was statistically significant (t =-21.308,P < 0.01).The results of pathological showed brain edema,rupture and eosinophilic and bilirubin deposition in the basal nuclei,and necrosis appeared in some severe parts.And there were different degrees of retardation and coordination disorders in the experimental group(s) newborn rhesus monkeys,but gradually returned to normal in 4 months later.Conclusion Intravenous injection of phenylhydrazine hydrochloride can be used to produce newborn rhesus monkey models of hemolytic hyperbilirubinemia.
6.Establishment and evaluation of a neonatal rat model of hyperbilirubinemia caused by hemolysis
Li YANG ; Baotian WANG ; Keying TIAN ; De WU ; Jiulai TANG
Acta Laboratorium Animalis Scientia Sinica 2015;(4):371-374
Objective To establish and evaluate a reliable and highly reproducible neonatal rat model of hyper-bilirubinemia and to provide an experimental basis for research of kernicterus and related mechanism of neuroinjury.Meth-ods Sixty 7-day old SD rats (28 male and 32 female) were used in this study.Three doses of phenylhydrazine hydrochlo-ride (25, 50, and 75 mg/kg) were intraperitoneally injected respectively to the neonatal rats to establish models of hyper-bilirubinemia induced by hemolysis.The control group was set up at the same time.48 hours after the experimental treat-ment, the bilirubin in blood and brain tissue, neuron-specific enolase (NSE) of brain tissue, and hemoglobin were detec-ted to evaluate the models.Results Compared with the control group, the bilirubin in the blood and brain tissue and the brain tissue NSE in the three experimental groups were significantly higher than that in the control group (P<0.05), while hemoglobin content was significantly lower than that in the control group (P<0.05).The bilirubin of blood and brain tis-sue and brain tissue NSE in the 50 mg/kg and 75 mg/kg dose phenylhydrazine hydrochloride groups were significantly high-er than that of the 25 mg/kg dose group ( P<0.05) , while hemoglobin content was significantly lower than that of the 25 mg/kg dose group ( P<0.05 ) .There were no significant differences between the 50 mg and 75 mg dose groups ( P>0.05).Conclusions Intraperitoneal injection of phenylhydrazine hydrochloride can be used to produce neonatal rat mod-els of hyperbilirubinemia, mimicking the clinical features of this disease, and 50 mg/kg of phenylhydrazine hydrochloride is the best concentration.It is an ideal method to establish newborn rat models of hyperbilirubinemia.
7.Establishment of SD neonatal rat hippocampal neuron models with human cytomegalovirus infection in vitro and preliminary study on calcium metabolism
Keyin TIAN ; De WU ; Li YANG ; Yuping CHEN ; Jiulai TANG
Chinese Journal of Applied Clinical Pediatrics 2014;29(6):455-458
Objective To investigate the influence of primary cultured neonatal rat hippocampal neurons caused by human cytomegalovirus (HCMV AD169) infection on intracellular calcium and its mechanism.Methods Twenty SPF SD rats born within 24 hours(10 cases of male and 10 cases of female) were assigned to establish the primary rat hippocampal neuronal monolayer cells; After cultured 8 days in vitro,the eligible cells were randomly divided into HCMV infection group,HCMV + MK-801 group,MK-801 group and control group,with 10 wells in each group.The fluorescence intensity values of the intracellular free calcium were detected after 24 hours of treatment with Fluo-3AM fluorescence staining.Results Inoculation of HCMV neurons after 24 h turned to round and swollen gradually,and 4days later,most of the cells disappeared; by immunohistochemistry in cultures of hippocampal neurons in HCMV,visible early proteins,brownish yellow granules,hematoxylin were found after being stained with brown pigment.The fluorescence intensity values of neuronal intracellular calcium (215.5 ± 14.9) in HCMV group was higher than that of control group (116.4 ± 5.9) (t =15.2,P < 0.01),whilerise,that in MK-801 group (88.1 ± 4.5) was significantly lower than that of control group,with decreased rate of (24.0 ± 6.7) % (t =-9.3,P < 0.01).The fluorescence intensity values of neuronal intracellular calcium in HCMV + MK-801 group (135.5 ± 8.6) was significantly decreased compared with that of HCMV group (215.5 ± 14.9),with decreased rate of (37.0 ± 3.4) % (t =11.3,P < 0.01).Conclusions Intracellular calcium overload of cultured rat hippocampal neurons in vitro with HCMV AD16 strains infection can be detected.One of its main mechanisms is the N-methyl-D-aspartic acid receptor channel-mediated calcium influx.
8.Preliminary Study on Serum Metabolites of Primary Biliary Cholangitis by Hydrogen Spectrum Nuclear Magnetic Resonance
Jing YANG ; De DUAN ; Huamei WU ; Lihong YANG ; Yan LI ; Jinhui YANG
Chinese Journal of Gastroenterology 2017;22(2):75-81
Hydrogen spectrum nuclear magnetic resonance (1 H-NMR ) is a commonly used method for metabolomics and has been applied in the study of liver cirrhosis and liver cancer,however,study on serum metabolites in patients with primary biliary cholangitis (PBC)is rare.Aims:To investigate the capability of 1 H-NMR for screening serum metabolites of PBC.Methods:Twenty PBC patients,20 HBV-related cirrhosis patients and 20 healthy controls were detected by 1 H-NMR.The 1 H-NMR spectra data were processed by principal component analysis (PCA)and orthogonal partial least squares-discriminant analysis (OPLS-DA)so as to create the diagnostic model.Based on the correlation coefficient P (corr),VIP value and non-paired t test of OPLS-DA model,the differential metabolites between normal group and the two cirrhosis groups were screened.Results:OPLS-DA model could effectively distinguish healthy controls and PBC patients,model interpretation ability and prediction ability were 81.9% and 44.8%,respectively (P=0.0293), glutamine,folic acid,urocanic acid,4-ethylbenzoic acid were differential metabolites.OPLS-DA model could also effectively distinguish healthy controls and HBV-related cirrhosis patients, urocanic acid, 1-methylhistamine, 1-methyladenine,glucose,L-acetylcarnitine were differential metabolites.OPLS-DA model could not effectively distinguish PBC patients and HBV-related cirrhosis patients.Conclusions:Serum glutamine and folic acid may be the potential biomarkers of PBC,which may be closely related to the immune damage mechanism and prognosis of PBC;1 H-NMR combined with OPLS-DA diagnostic model are expected to become a new method for studying liver cirrhosis.
9.Study of interaction between NS3 serine protease of HCV and wild type P53
Wu, OU ; E-De, QIN ; Cui-hong, YANG ; Pei-ying, YANG ; Man, YU
Bulletin of The Academy of Military Medical Sciences 2001;25(1):21-23
Objective:To investigate the molecular interaction between non-structural protein 3 serine protease of hepatitis C virus(HCV)and wild type P53,and to lay the basis for elucidating the mechanism of oncogenesis of hepatocellular carcinoma(HCC)after infection of HCV.Methods:The recombinant plasmids,pGAD424-NS3,pGAD424-NS315aa- and pGAD424-NS330aa-,were constructed and the interaction between NS3 serine protease and its cofactor NS4A,the interaction between wild type P53 and NS3 serine protease and its N-truncated mutants were dectected qualitatively and quantitatively in yeast two-hybrid system.Results:The results indicated that interaction existed not only between full-length NS3 serine protease and P53,but also between N-truncated mutants of NS3 serine protease and P53.Furthermore,the difference between enzyme activity unit(IU)of β-gal induced by these interactions was not significant(P>0.05).Conclusions:NS3 serine protease of hepatitis C virus and its N-truncated mutants can interact with wild type P53,and the region of NS3 serine protease involved in the interaction may be located in its C-terminal,but not in its N-terminal.
10.Magnetic resonance imaging characteristics of spinal cord lesions and brain abnormalities in Chinese patients with neuromyelitis optica.
Lei WU ; Yang YANG ; De-hui HUANG ; Hui XIAO ; Wei-ping WU
Journal of Southern Medical University 2010;30(12):2733-2737
OBJECTIVETo analyze the magnetic resonance imaging (MRI) characteristics of neuromyelitis optica (NMO) in Chinese patients.
METHODSWe retrospectively reviewed the MRI films of 61 patients with NMO (including 57 female and 4 male patients) admitted in our department.
RESULTSOf these patients, 39 (79.6%) showed positivity for serum aquaporin-4 (AQP4) antibody. On MRI, 18 patients showed involvement of the cervical cord alone, 27 had both cervical and thoracic segment involvement, and 16 displayed thoracic segment involvement. The lesions appeared linear (9 cases), diffuse (23 cases), or both (29 cases), mostly located axially with occasional lateral distribution. Thirty-nine of the 61 patients (63.9%) had brain abnormalities, 31 presented with supratentorial lesions (mostly in the juxtacortical, subcortical, deep white matter and lateral ventricle-adjacent regions, n=27), 15 showed infratentorial lesions (mostly in the preiaqueduct-fourth ventricular-central canal, n=13), and 7 had supra- and infratentorial lesions simultaneously.
CONCLUSIONNMO has complex MRI presentation, and linear lesions in the spinal cord and preiaqueduct-fourth ventricular-central canal lesions, where AQP4 is high expressed, can be characteristic for NMO. MRI and AQP4 antibody detection are suggested for suspected cases for early diagnosis.
Adult ; Aquaporin 4 ; immunology ; Brain ; pathology ; Female ; Humans ; Magnetic Resonance Imaging ; Male ; Neuromyelitis Optica ; pathology ; Retrospective Studies ; Spinal Cord ; pathology