● AIM: To investigate the sequential changes of HIF-1 α Protein and mRNA in hypoxic bovine retinal microvessel endothelial cells.● METHODS: The bovine retinal microvessel endothelial cells were cultured in normoxic and CoCl2-induced hypoxic conditions respectively. Expressions of HIF-1 α Protein were measured with immunohistochemical staining, and RT-PCR was used to determine the HIF-1 α mRNA.● RESULTS: HIF-1 α began to increase 1h after hypoxia,and reached the peak at 4h. After 16h, it declined significantly. Compared with the normoxic group, the expression of HIF-1 α protein in the hypoxic groups had significant difference (P＜0.01), and HIF-1 α mRNA expression was unchanged under hypoxia.● CONCLUSION: HIF-1 α participates in the hypoxic procedures in retinal microvessel endothelial cells, and hypoxia induce time-dependent changes of HIF-1 α protein expression, which is not modulated on the transcription level. Analysis of HIF-1 α expression revealed a temporal and spatial changes with regard to the hyperoxic repression, indicating that HIF-1 may play a major role in the development of retinopathy of prematurity and other ischemic retinal disorders such as diabetic retinopathy.

Objective To evaluate the biophysical properties of insulin-producing cells labeled and unlabeled with superparamagnctic iron oxide nanoparticals and poly-L-lysine(SPIO-PLL)in vitro,and then monitor cellular imaging with 1.5 T MR.Methods BMSCs were isolated from tibia and femur of 6～8 weeks normal Spragne-Dawley rats,purified on the basis of their ability to adhere to the matrix,and expanded through their self-renewal.Two-step strategy was adopted with BMSCs induced into insulin-producing cells, After that,the cells were incubated with SPIO-PLL.Prussian blue stain was employed for identifying intracellular irons.Radioimmunology assay was used to measure the insulin secretion by the labeled and unlabeled cells,and later on underwent MR imaging with T_1WI、T_2WI、T_2*WI sequences,Results lntracytoplasmic nanoparticales were stained with Prussian blue possessing insulin-producing cells labeled with SPIO-PLL.The amount of insulin secreted by the labeled and unlabeled ceils had no statistical significant difference.The signal intensity of labeled cells decreased significantly on T_2*WI,as well as the stronger proportional variations for signal intensity.Conclusion Insulin-producing cells can be labelled effectively with SPIO-PLL and be imaged by 1.5 T MRI.(J Intervent Radiol,2007,16:104-108)

OBJECTIVETo establish a set of suitable and reliable methods for HBV genotyping and to study the distribution of HBV genotypes.

METHODSType-specific nucleotides were searched through alignment of S genes (more than 1000 sequences) listed in GenBank. Then, type-specific primers were designed and type-specific primer PCR was used to genotype the 238 HBV strains. S genes of the untyped strains were further amplified and sequenced to find out their genotypes with type-specific nucleotide analysis.

RESULTSAll the 238 HBV strains were genotyped. 159 (66.8%) cases were genotype B, 69 (28.9%) were genotype C, 6 (2.5%) were mixtures of genotypes B and C and 4 (1.6%) were mixtures of genotypes B and D. No genotypes of A, E, F, G, and H were found.

CONCLUSIONGenotypes B and C are the most common types for HBV strains. Mixtures of genotypes B and C or genotypes B and D coinfection rarely existed. There is no relationship between the gender of the patients and HBV genotypes (X2 = 0.794, P more than 0.05).

DNA Primers ; DNA, Viral ; blood ; genetics ; Female ; Genotype ; Hepatitis B virus ; genetics ; Hepatitis B, Chronic ; virology ; Humans ; Male ; Nucleotides ; genetics ; Polymerase Chain Reaction ; methods ; Sequence Analysis, DNA

Objective To explore the feasibility of mobilization circulating MSCs by G-CSF and observe the repairing effect of G-CSF mobilization in severe mouse traumatic brain injury(TBI) model.Methods MSCs-derived bone marrow and peripheral blood(PB) were cultured and its CFU-F were counted after mobilization by G-CSF.At 2,24,48,96,120,144,192,264,336 h after severe TBI in mice was establish,the neurobehavior of mice was measured by neurological examination and motor functional test,and mortality rate and pathologic changes were analyzed.Results MSCs-derived PB were successfully cultured.The CFU-F of mobilization group increased significantly than that of control group(P

Objective To invesgate the effect of P-glycoprotein(PGP)inhibitor,verapamil,on electrobiological activity and seizure behavior in phenytoin-carbamazepine(PHT-CBZ)resistant rats.Methods The model of medically intractable epilepsy was established by kindling of amygdale. Verapamil was applied to PHT-CBZ resistant rats,followed by the observation on after discharge threshold (ADT),after discharge duration(ADD)and seizure activity.Results Compared with the control group, the ADT was higher in PHT-CBZ resistant rats peritoneally injected with verapamil((238.0?32.2)?A vs (177.0?23.3)?A,P

OBJECTIVETo investigate the development and maturation competence of oocytes retrieved from cryopreserved and transplanted human fetal ovarian tissue by techniques of tissue culture, inducing ovary, oocyte retrieval, and in vitro maturation (IVM).

METHODSFetal ovaries of 20 weeks were frozen-thawed and cultured for 6 days in vitro, then xenografted into kidney capsules of immunodeficient mice. All mice were stimulated with follicle stimulating hormone every second day for 23 weeks, starting 1 week after grafting. Then oocytes were retrieved from antral follicles 13 hours after human chorionic gonadotrophin injection. IVM was performed to evaluate the maturation competence of the oocytes from ovarian grafts. Human fetal ovarian tissues were examined with histological and proliferating cell nuclear antigen (PCNA) evaluation.

RESULTSThere was no difference between fresh ovarian tissues and frozen-thawed ovarian tissues in the percentage of follicles at different growth stages (P > 0.05). The proportion of the primary follicles and preantral follicles in the cultured ovarian tissues was significantly larger than that of fresh ovarian tissues and frozen-thawed ovarian tissues (P < 0.05). The proportion of the primary follicles, preantral follicles, and antral follicles in the transplanted ovarian tissues was significantly higher than that of cultured ovarian tissues, fresh ovarian tissues, and frozen-thawed ovarian tissues (P < 0.05). No significant signals of PCNA in the primordial follicles in all ovarian tissues were observed. PCNA immunoreactivity first appeared in primary follicles. However, the obviously positive signals of PCNA were seen in the oocytes and/or the granular cells of cultured ovarian tissues and transplanted ovarian tissues. Oocytes from antral follicles were collected and matured in vitro, and 21.43% of the oocytes reached to MII within 48 hours IVM.

CONCLUSIONSHuman ovarian follicles can survive and develop well after cryopreservation, tissue culture, and xenotransplantation. Furthermore, oocytes recovered from grafts have normal maturation competence.

Animals ; Cryopreservation ; Female ; Humans ; Mice ; Mice, Inbred BALB C ; Oocytes ; cytology ; Oogenesis ; Ovarian Follicle ; cytology ; growth & development ; transplantation ; Pregnancy ; Transplantation, Heterologous

China ; epidemiology ; Hand, Foot and Mouth Disease ; epidemiology ; Humans ; Incidence ; Weather

Objective To investigate the changes of morphological structures and biomechanical properties of scleral tissues in rabbits at different month ages. Methods The eyeballs of 1, 2 and 3 month-old New Zealand white rabbits were obtained for measuring the diameter and axial length, and the thickness of scleral tissues. Part of the scleral tissues was used to observe scleral structures with HE staining, some other part was used to observe collagen fibrils by electron microscope, and the left part were cut into strips and used to test the elastic modulus of the sclera on Instron 5544 system. Results The diameter, axial length of eyeballs and the thickness of the sclera were increased with month age. The elastic modulus of scleral tissues was also increased with month age. The numbers of scleral fibroblasts were decreased and the numbers of fiber bundles were increased with month age. The diameters of collagen fibrils were increased with month age. Conclusions In the post-embryonic stages, the structures of eyeball and sclera are changed continually, with growing numbers of thicker collagen fibers, and the biomechanical properties of scleral tissues are improved correspondingly. The mechanism of post-embryonic development in sclera is further explained in the study, which can provide theoretical guidance for prevention of sclera-related diseases.

OBJECTIVEThe paper is a study on the clinical symptoms and pathogeny of ectrodactyly and absence of radius side part palm and split foot malformation of some patients in one family.

METHODSBased on the patient family investigation,a normal control group and a patient group were established. Then, polymerase chain reaction technique was used for DNA sequencing and analysis of the two groups for their exons 5-8 gene group DNA of P63 gene.

RESULTSThe medical examination found that the patients' upper bilateral limbs are short of thumbs, forefingers and middle fingers, and have radius side part palm and double lower limbs foot clefts malformation. The pathogeny research revealed that the PCR expansion pieces of the exons 5-8 of P63 are 284 bp, 259 bp, 245 bp and 259 bp respectively, and the size of the expansion piece of the patients was the same as that of the normal people group. However, a respective comparison between the DNA serial of the expansion piece of the patient and that of the normal people group and that of the P63 gene in the human gene bank showed that mutation occurs at the number 665 base pair of exon 5 of P63, namely a mutation from G to A.

CONCLUSIONThe ectrodactyly, absence of radius side part palm and split foot malformation are caused by the mutation of base pair at number 665 of the exon 5 of P63.

Exons ; genetics ; Female ; Foot Deformities, Congenital ; genetics ; pathology ; Genetic Predisposition to Disease ; Hand Deformities, Congenital ; genetics ; pathology ; Humans ; Male ; Membrane Proteins ; genetics ; Mutation ; Pedigree ; Polymerase Chain Reaction

OBJECTIVETo explore the mechanism for adefovir dipivoxil (ADV) resistance occurred in chronic hepatitis B patients of a series of phase III clinical trails.

METHODS30 resistant HBV strains were selected out from 177 cases of ADV treated chronic hepatitis B patients. HBV polymerase RT region were amplified by nested PCR and analyzed with the standard nucleotide sequence of HBV strains deposited in GeneBank.

RESULTS21 out of 30 HBV strains were primary resistant strains, among them 5 HBV strains (23.8%, 5/21) had the polymorphism site of rtN118H. While the other 9 HBV strains showed secondary resistance, variations in conservative region C (rtM207V) and other non-conservative regions were found. The classic mutation sites such as rtN236T and rtA181V/T were not found.

CONCLUSIONSPolymorphism site of rtN118H might be responsible for HBV primary resistance to ADV therapy. rtM207V variation in HBV RT C domain and other variation sites might play a role in HBV secondary resistance to ADV treatment, and natural resistant quasispecies may be the basis for the ADV quick resistance. These conclusions await further confirmation by phenotype test.

Adenine ; analogs & derivatives ; pharmacology ; therapeutic use ; Adult ; Alanine Transaminase ; blood ; Amino Acid Sequence ; Antiviral Agents ; pharmacology ; therapeutic use ; Base Sequence ; DNA Primers ; DNA, Viral ; blood ; Drug Resistance, Viral ; Female ; Genotype ; Hepatitis B virus ; drug effects ; genetics ; Hepatitis B, Chronic ; drug therapy ; genetics ; virology ; Humans ; Male ; Molecular Sequence Data ; Organophosphonates ; pharmacology ; therapeutic use ; Polymorphism, Genetic ; genetics ; RNA-Directed DNA Polymerase ; drug effects ; genetics ; Reverse Transcriptase Inhibitors ; pharmacology ; therapeutic use ; Reverse Transcriptase Polymerase Chain Reaction ; methods ; Sequence Analysis, DNA