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 .

Objective: To construct recombinant adenovirus vector of Antisense BTEB2 cDNA and study the effect of antisense BTEB2 on proliferation of vascular smooth muscle cells and neointimal hyperplasia.Methods:BTEB2 cDNA was prepared by RT PCR technique and was subcloned reversedly into shuttle plasmid pDC315 to constructed recombinant shuttle plasmid pDC315AS BTEB2.Then the recombinant shuttle plasmid and adenovirus genomic plasmid pBHGlox?E1,3Cre were cotransfected into 293 cell to obtain recombinant adenovirus. The PCR technique was used to detect target gene fragment and adenovirus genomic characteristic fragment. After the recombinant adenovirus infected vascular smooth muscle cells, antisense RNA expression of BTEB2 was detected by RT PCR. Results:There was recombinant adenovirus containing BTEB2 cDNA in the lysate of cotransfected 293 cells.The recombinant adenovirus infected 293 cells and replicated in 293 cells. The expression of BTEB2 antisense RNA was very obvious in vascular smooth muscle cells after infected by recombinant adenovirus. Conclusion:The construction of recombinant adenovirus vector of Antisense BTEB2 has been achieved, and Antisense RNA of BTEB2 can express in vascular smooth muscle cells. This study has paved the way for furthur research.

Objective To construct the recombinant adenovirus of Angiotensin Ⅱ type 2 receptor (AT2R), and study the effect of AT2R on prevention of neointimal hyperplasia in rat carotid arteries after balloon angioplasty. Methods The recombinant adenovirus of AT2R with green fluorescent protein was constructed by using the method of homogenous recombination in bacteria. AT2R gene was transduced into rat carotid arteries with pAdCMV/AT2R after the reproduction of rat carotid balloon injury restenosis model. The expression and the transfection efficiency of AT2R gene were evaluated by RT-PCR, immunofluorescence staining, and confocal microscope. The intimal/medial area ratio was measured by digital analysis system. Results The titre of purified recombinant adenovirus was 1.5?10 12pfu/ml, and it was proved by PCR. pAdCMV/AT2R transfection efficiency in carotid artery was about 40% at day14, and localized to the neointima, as well as to the media and the adventitia. pAdCMV/AT2R delivered into injured rat carotid arteries significantly up-regulated AT2RmRNA and protein expression in neointima from day 7 to 21 after injury. Compared with pAd-GFP transfection group, pAdCMV/AT2R transfection reduced I/M ratio significantly on day 21(0.78?0.06 vs. 1.36?0.21 respectively, P

Objective To study the effect of rat angiotensin Ⅱ receptor (AT2R) gene transfer mediated by adenoviral vector on neointimal proliferation after rat carotid artery injury. Methods AT2R gene was transducted into the carotid artery by adenoviral vector with GFP after carotid balloon injury. Restenosis models were established in Wistar rats. Immunostaining was performed to examin the expression of AT2R in local arteries. Neointima/media area ratio at day 21 after gene transfer was measured by morphometric analysis. Results The transduction rate of AT2R gene in rat carotid arteries was 40% and the gene expressed constantly in the vessel walls. Neointima/media area ratio of the gene transduction group was significantly reduced (48%) compared with the control group at day 21 after gene transfer. Conclusion The results suggest the possibility that AT2R gene transfer can inhibit proliferation of arterial intima after balloon injury and it is a potential therapeutic approach to prevent neointimal hyperplasia.

OBJECTIVETo analyze the clinical effects of short-segment posterior pedicle screw combined with transpedicle vertebral bone grafting in treating thoracolumbar burst fractures.

METHODSFrom March 2008 to March 2013, 62 patients with thoracolumbar burst fractures were treated with short-segment posterior pedicle screw combined with transpedicle vertebral bone grafting. Including 40 males and 22 females, the age from 17 to 65 years old with an average of 38 years. According to AO classification, 34 cases were type A3.1, 7 cases were type A3.2 and 21 cases were type A3.3. Load-sharing scores were from 4 to 6 points with an average of 5.4 points. According to ASIA grade, 2 cases were grade C, 5 cases were grade D and 55 cases were grade E. Preoperative, postoperative at 3 d and final follow-up, the Cobb angle, the relative height of anterior vertebral body and the encroachment rate of spinal canal were measured by X-ray films and computed tomography (CT) scan, meanwhile, the information of bone healing and spinal nerves recovery were observed.

RESULTSAll patients were followed up from 11 to 14 months with an average of 12.2 months. The duration of removing internal fixation were from 9 to 13 months (averaged, 11.5 months). One suffered from infection and was cured by debridement. Two cases had mild pain of back. At 6 months after operation, according to ASIA grade to evaluate never function, 1 case was grade C, 3 cases were grade D and 58 cases were grade E. X-ray and CT showed the fractures obtained good union at final follow-up. The Cobb angle, the relative height of anterior vertebral body and the encroachment rate of spinal canal had obviously improved at 3 days after operation (P<0.05); but there was no significant differences between postoperative at 3 d and final follow-up (P>0.05).

CONCLUSIONShort-segment posterior pedicle screw combined with transpedicle vertebral bone grafting is an effective method to treat thoracolumbar burst fractures. It can reduce the loss of postoperative correction and prevent the internal fixation failure.

Adolescent ; Adult ; Aged ; Bone Transplantation ; Female ; Fracture Fixation, Internal ; methods ; Humans ; Lumbar Vertebrae ; injuries ; surgery ; Male ; Middle Aged ; Pedicle Screws ; Spinal Fractures ; surgery ; Spinal Fusion ; Thoracic Vertebrae ; injuries ; surgery

AIM: To study the effects of BTEB2 antisense RNA on the proliferation of vascular smooth muscle cells(VSMC) and its mechanism. METHODS: After the recombinant adenovirus Ad.ASBTEB2 infected VSMC, antisense RNA and protein expression of BTEB2 were evaluated respectively by RT-PCR and Western blotting. Proliferation and cell cycle progress of VSMC were analyzed respectively by MTT test and flow cytometry. The expression of PCNA, AT1R and PDGF-BB were detected by immunocytochemistry. RESULTS: The expression of BTEB2 antisense RNA was demonstrated in VSMC after infected by recombinant adenovirus. Ad.ASBTEB2 infection significantly inhibited BTEB2 protein expression and proliferation of VSMC induced by serum, and resulted in G_0/G_1 blocking. The inhibitory effects of BTEB2 antisense RNA on the expression of PCNA, AT1R and PDGF-BB were demonstrated by immunohistochemistry. CONCLUSION: BTEB2 antisense RNA significantly inhibits the proliferation of VSMC, probably by suppressing the expression of VSMC proliferation related genes, such as PCNA, AT1R and PDGF-BB.

Objective To construct recombinant adenoviral vector expressing basic transcription element binding protein 2 (BTEB2) antisense RNA and study the effects of BTEB2 antisense RNA on the proliferation of vascular smooth muscle cells (VSMCS). Methods BTEB2 cDNA was prepared by RT-PCR technique and was subcloned reversedly into the shuttle plasmid to construct recombinant shuttle plasmid, and then the recombinant shuttle plasmid and adenovirus genomic plasmid pBHG were cotransfected into 293 cells to obtain the recombinant adenovirus. The PCR technique was used to detect the target gene fragment and adenovirus genomic characteristic fragment. After transfection of the recombinant adenovirus into VSMCs, BTEB2 antisense RNA was detected by RT-PCR. The effects of BTEB2 antisense RNA on BTEB2 protein expression, proliferation, and cell cycles of VSMCs were analyzed by Western blot, MTT test, and flow cytometry, respectively. Results The recombinant adenovirus was proved correct by PCR. The expression of BTEB2 antisense RNA was very obvious in VSMCs after infection by recombinant adenovirus. Recombinant adenovirus infection significantly inhibited BTEB2 protein expression and proliferation of VSMCs and resulted in G 0/G 1 block. Conclusion The recombinant adenoviral vector expressing BTEB2 antisense RNA has been constructed. BTEB2 antisense RNA can significantly inhibit the proliferation of VSMCs.

Objective:To construct the recombinant adenovirus of AT2R by using the method of homogenous recombination in bacteria. Methods:AT2R gene was get from the vector of PUHD-AT2R by PCR ,and subcloned into shuttle vector of pAdTrack-CMV, forming transfer vector of pAdTrack-CMV/AT2R.Then it was linearized with PmeⅠ and transformed into Adeasier-1 cell. The DNA of identified recombinant plasmid was digested with PacⅠ and transfected to 293cells to package adenovirus. The PCR technique was used to detect target gene . The titre of the Ad-AT2R was measured with the aid of GFP expression. Results:PCR test indicated each the recombinant adenovirus contained the insert of AT2R. The titre of purified recombinant adenovirus was 1.5?10 12pfu/ml. Conclusion:The method of homologous recombination in bacteria is more convenient and efficient compared with that in cell .The prepared Ad-AT2R paves a sound foundation for further study.

In the 1960s, modern science began involving the essence of heat syndrome, but there have still no in-depth systematic studies on pathological mechanisms of heat syndrome and action mechanisms of cold and cool herbs. In this study, the animal model with heat syndrome was set up by feeding herbs with hot property, and then cold and cool herbs was applied in the experimental therapy. The two-dimensional electrophoresis and mass spectrometry technologies were adopted to compare the liver mitochondria proteome of the rats of the heat syndrome model and the ones treated with cold and cool herbs, so as to discover specificity-related proteins after heat syndrome and treatment with cold and cool herbs.
Animals ; Carbohydrate Metabolism ; drug effects ; Cold Temperature ; Drugs, Chinese Herbal ; pharmacology ; Energy Metabolism ; drug effects ; Hot Temperature ; Lipid Metabolism ; drug effects ; Male ; Mitochondria, Liver ; drug effects ; metabolism ; Proteome ; metabolism ; Rats ; Rats, Sprague-Dawley

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.