Objective To investigate the effects of FKBP12.6 gene transfection on the contractility of ventricular myocytes of rats with heart failure.Methods Rat model of heart failure was established by a surgical operation of abdominal aortic stenosis.The enzymatic hydrolysis was employed to isolate the rats' ventricular myocytes.Mediated by adenovirus the FKBP12.6 gene was transfected into ventricular myocytes.Ad-GFP infected cardiac myocytes from heart failure rats and normal rats were used as control.Western blotting and reverse transcriptase-polymerase chain reaction(RT-PCR) were used to detect the specific overexpression of FKBP12.6.Transfected ventricular myocytes were loaded with the membrane-permeant Ca2+ dye X-rhod-1-AM.Under the condition of field stimulation,the laser confocal microscope in line-scan and plane-scan mode was used to detect the Ca2+ transients and myocytes contraction.Results The mRNA and protein levels of FKBP12.6 in the myocytes of rats with heart failure were significantly lower than that in normal myocytes(0.47?0.08 vs 0.96?0.12,0.25?0.04 vs 0.48?0.07,P

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 .

Humans ; Infant ; Male ; Mandibulofacial Dysostosis

Adult ; Female ; Hematopoietic Stem Cell Transplantation ; Humans ; Lymphoma, Extranodal NK-T-Cell ; surgery ; Male ; Middle Aged ; Retrospective Studies ; Young Adult

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.

Objective To observe the inhibitory effects of local co-transfection of tissue-type plasminogen activator(tPA) gene and proliferating cell nuclear antigen antisense oligodeoxynucleotides(PCNA-ASODN) on the intima proliferation and restenosis of autograft artery in rabbits. Methods One hundred and twenty male Zelanian rabbits were randomly divided into four groups(n=30, in each group): control group, PCNA-ASODN group, tPA group and tPA+PCNA-ASODN group. The left and right external iliac arteries (length 1.0 cm) were transplanted reciprocally. The transplanted arteries were respectively soaked in lipofection, PCNA-ASODN, pBudCE4.1/tPA and pBudCE4.1/tPA+PCNA-ASODN solution about 15 minutes. The transplanted arteries were sutured with 9-0 sutures soaked in PCNA-ASODN and pBudCE4.1/tPA solution. Each group were divided into five subgroups(n=6, in each subgroup) according to the sacrifice time (3 d, 7 d, 14 d, 28 d and 56 d after operation). On every sacrifice time point, the vascular specimens were harvested. The thrombocyte assembling and thrombus forming lining vessel wall were observed by scanning electron microscope. The pathological morphology of transplanted arteries were observed under microscope(HE). The intimal areas and stenosis ratio(%) of transplanted arteries were calculate and analyzed statistically among groups by computer system. The mRNA expression of tPA gene in transplanted ressel wall was detected with vevere transcription-PCR(RT-PCR). The number of PCNA positive cells in transplanted vessel wall was counted by SP immunochemisty. Results The mRNA expression of tPA gene in the transplan-ted vessel wall in tPA and tPA+PCNA-ASODN groups was higher than that of the other two groups (P

AIM: To observe the effects of local transfection of vascular endothelial growth factor 165 (VEGF165) gene on inhibiting intimal hyperplasia and restenosis of artery in rabbits after operation injury, and its possible mechanisms. METHODS: Microsurgery injury was used to establish the intimal injury model of right external iliac artery in rabbits. 105 male New Zealand rabbits were randomly divided into 3 groups (35 rabbits in each group). Group A was physiological saline control group, group B was pBudCE4.1-transfected group, group C was pBudCE4.1/VEGF165-transfected group. The physiological saline, pBudCE4.1 and pBudCE4.1/VEGF165 transfection solutions were injected into injured vessel walls of above-mentioned groups. The injured vascular specimen was harvested for pathologic examination, electric microscope observation, RT-PCR examining and immunohistochemical staining. RESULTS: Rabbit intimal thickness and area of vessel walls in group C at every time point after operation were significantly less than those in group A and group B (P

AIM:To study the effects of FK506 binding protein 12.6(FKBP12.6) overexpression on the performance of sarcoplasmic reticulum(SR) in ventricular myocytes of rats with heart failure.METHODS:The adenovirus(Ad)-mediated gene transfer was used to overexpress FKBP12.6 in ventricular myocytes of rats with heart failure.Western blotting and reverse transcriptase-polymerase chain reaction(RT-PCR) analysis were used to reveale specific overexpression of FKBP12.6.X-rhod-1-AM was used as the Ca2+ indicator,and cells were viewed under a confocal microscope.RESULTS:Adenovirus mediated overexpression of FKBP12.6 resulted in a 5-fold increase in relative FKBP12.6 mRNA levels and a 4-fold increase in relative FKBP12.6 protein levels at 48 h after transfection compared with control.The amplitude of the fluorescence [Ca2+]i transient was significantly increased in Ad-FKBP12.6-GFP cardiomyocytes compared with Ad-GFP myocytes(peak F/F0,3.16?0.42 vs 1.43?0.38,P

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.

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.