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Biomedical subjects

Jing Hu

Publications and source records attributed to Jing Hu.

84 records · Page 5Linked to original sources

Novel TTV variants isolated in an epidemic of hepatitis of unknown etiology.

TT virus (TTV) is a recently discovered single-stranded DNA virus that has been reported to be associated with elevated transminase levels in the patients with posttransfusion hepatitis of unknown etiology. TTV prevalence is very high in the common population and its pathogenicity remains unclear. In this study, we performed an epidemiological study to investigate the infection rate of TTV and its role in an epidemic of unknown-etiology hepatitis. Moreover, two TTV isolates named L01 and L02 were cloned from the serum of a patient with unknown-etiology hepatitis. Eighty-one subjects were included in the study and were divided into two groups: 18 in the case group and 63 in the control group. TTVDNA was detected by nested PCR from sera samples. The infection rates of TTV in case and control groups were 33.3 and 38.9%, respectively. There was no significant difference between the two groups. Homology analysis showed that L01 had a very poor homology with other TTV isolates and L02, and L02 was 75.5% identical to JA10. The result does not support TTV as a causative agent in this epidemic. The genetic divergence between L01 and other TTV isolates beyond genotype, so it represents a new genotype of TTV.

China↗

Fibronectin receptor reduction in skin and fibroblasts of patients with Ullrich's disease.

Ullrich's disease is a congenital muscular dystrophy characterized clinically by generalized muscle weakness, multiple contractures of the proximal joints, and hyperextensibility of the distal joints. Recent studies have demonstrated that collagen VI is deficient in the muscles of patients with Ullrich's disease, and some cases result from recessive mutations of the collagen VIalpha2 gene (COL6A2). Fibronectin is one of the main components of the extracellular matrix (ECM) and associates with a variety of other matrix molecules including collagen. The behavior of fibronectin on cells is mediated by fibronectin receptors, members of the integrin family. We studied the expression of fibronectin receptors and fibronectin in patients with Ullrich's disease, and found a marked reduction of fibronectin receptors in the ECM of skin and cultured fibroblasts of these patients. These results suggest that collagen VI deficiency may lead to the reduction of fibronectin receptors and that an abnormality of cell adhesion may be involved in the pathogenesis of Ullrich's disease.

Cells, Cultured↗

Synthesis, biodegradability, and biocompatibility of lysine diisocyanate-glucose polymers.

The success of a tissue-engineering application depends on the use of suitable biomaterials that degrade in a timely manner and induce the least immunogenicity in the host. With this purpose in mind, we have attempted to synthesize a novel nontoxic biodegradable lysine diisocyanate (LDI)- and glucose-based polymer via polymerization of highly purified LDI with glucose and its subsequent hydration to form a spongy matrix. The LDI-glucose polymer was degradable in aqueous solutions at 37, 22, and 4 degrees C, and yielded lysine and glucose as breakdown products. The degradation products of the LDI-glucose polymer did not significantly affect the pH of the solution. The physical properties of the polymer were found to be adequate for supporting cell growth in vitro, as evidenced by the fact that rabbit bone marrow stromal cells (BMSCs) attached to the polymer matrix, remained viable on its surface, and formed multilayered confluent cultures with retention of their phenotype over a period of 2 to 4 weeks. These observations suggest that the LDI-glucose polymer and its degradation products were nontoxic in vitro. Further examination in vivo over 8 weeks revealed that subcutaneous implantation of hydrated matrix degraded in vivo three times faster than in vitro. The implanted polymer was not immunogenic and did not induce antibody responses in the host. Histological analysis of the implanted polymer showed that LDI-glucose polymer induced a minimal foreign body reaction, with formation of a capsule around the degrading polymer. The results suggest that biodegradable peptide-based polymers can be synthesized, and may potentially find their way into biomedical applications because of their biodegradability and biocompatibility.

Biocompatible Materials↗

Characterization of a putative alpha-helix across the capsid-SP1 boundary that is critical for the multimerization of human immunodeficiency virus type 1 gag.

A 14-amino-acid spacer peptide termed SP1 that separates the capsid (CA) and nucleocapsid (NC) sequences plays an active role in the assembly of human immunodeficiency virus type 1. This activity of SP1 involves its amino-terminal residues that, together with adjacent CA residues, constitute a putative alpha-helical structure spanning Gag residues from positions 359 to 371. In this study, we have determined that the virus assembly determinants within this putative alpha-helix were residues H359, K360, A361, L364, A367, and M368, of which K360 and A367 contribute to virus production to lesser extents. Notably, changes of the two basic amino acids H359 and K360 to arginine (R) impaired virus production, whereas mutations L364I and M368I, in contrast to L364A and M368A, generated near-wild-type levels of virus particles. This suggests that within Gag complexes, amino acids H359 and K360 are involved in stricter steric interactions than L364 and M368. Since L364 and M368 are separated by four residues and thus presumably located on the same side of the helical surface, they may initiate synergistic hydrophobic interactions to stabilize Gag association. Further analysis in the context of the protease-negative mutation D185H confirmed the key roles of amino acids H359, A361, L364, and M368 in virus assembly. Importantly, when transfected cells were subjected to Dounce homogenization and the cell lysates were treated by ultracentrifugation at 100,000 x g, Gag molecules containing each of the H359A, A361V, L364A, and M368A mutations were found mainly in the supernatant fraction (S100), whereas approximately 80% of wild-type Gag proteins were found in the pellet. Therefore, these four mutations must have prevented Gag from generating large complexes.

Amino Acid Sequence↗

Expression of midkine in regenerating skeletal muscle fibers and cultured myoblasts of human skeletal muscle.

Midkine is a member of a family of developmentally regulated neurotrophic and heparin-binding growth factors. The expression of midkine was examined immunohistochemically in biopsied muscle specimens from patients with various myopathies. The intense immunoreactivity of midkine was observed in the sarcoplasm at an early stage regenerating fibers of small diameter. Midkine immunoreactivity was also detected in cultured human skeletal muscle myoblasts from the proliferation stage to the differentiation stage. These results suggest that midkine may be an important skeletal muscle growth-promoting factor that participates in the growth process during skeletal muscle development, proliferation, and differentiation, and in the skeletal muscle regeneration in various neuromuscular disorders. This is the first report demonstrating the presence of midkine in skeletal muscle cells.

Biopsy↗

Selective p38 activation in human non-small cell lung cancer.

The mitogen-activated protein kinase (MAPK) pathways transmit signals from the cell membrane to the nucleus. Activation of MAPK cascades may play a role in malignant transformation. We hypothesized that enhanced expression of one or more of these pathways would occur in human lung cancers. Using Western blot analysis of tissue homogenates from resected non- small cell lung cancers and matched non-neoplastic lung tissue, we determined that only activated p38 was consistently increased in tumor compared with normal tissue. In vitro kinase assays confirmed that the levels of activated MAPK correlated with the activity of the enzymes, and immunohistochemical analysis confirmed the cellular localization of the activated MAPKs. We incubated a lung cancer cell line in a hypoxic chamber to simulate the hypoxic environment in solid lung tumors, but found no increase in p38 activation. Contrary to our expectations, ERK and JNK, the MAPK pathways traditionally associated with cell growth and perhaps malignant transformation, were not consistently activated in the human lung tumor samples. However, p38, a MAPK usually associated with stress responses, growth arrest, and apoptosis, was activated in all of the human lung cancer samples, suggesting an additional role for this pathway in malignant cell growth or transformation.

Blotting, Western↗

Glucocorticoids inhibit lung cancer cell growth through both the extracellular signal-related kinase pathway and cell cycle regulators.

Glucocorticoids inhibit the proliferation of various cell types, but the mechanism of this inhibition remains unclear. We investigated the effect of dexamethasone on non-small cell lung cancer cell growth and cell cycle progression. We showed that dexamethasone suppresses the proliferation of A549 and Calu-1 cells, with accumulation of cells in G1/G0 stage of the cell cycle, as determined by fluorescence-activated cell sorter analysis. Western blot analysis confirmed that this is associated with hypophosphorylation of retinoblastoma protein. Using Western blot analysis and in vitro kinase assays, we found that dexamethasone results in decreased activity of CDK2 and 4, decreased levels of cyclin D, E2F, and Myc, and increased levels of the CDK inhibitor p21(Cip1). In addition, we found that dexamethasone decreases activity of extracellular signal-related kinase (ERK)/mitogen-activated protein kinase (MAPK). The kinetics of all these changes indicate that inhibition of the ERK/MAPK pathway precedes the cell cycle effects, suggesting that regulation of this MAPK-signaling pathway may be an alternative mechanism for glucocorticoid-induced cell cycle arrest and growth inhibition.

Cell Cycle↗

Biomechanical properties of regenerated bone by mandibular distraction osteogenesis.

OBJECTIVE: To study the biomechanical properties of the new bone generated by mandibular distraction osteogenesis (DO). METHODS: A total of 11 healthy adult goats were randomly divided into 2 groups, the experimental group (n=9) and the control group (n=2). For the goats in the experimental group, the bilateral mandibles were gradually lengthened for 10 mm with distraction appliances. Three goats were sacrificed respectively at 2, 4 and 8 weeks after completion of distraction. Compressive, three-point bending and shearing tests were conducted on the standard regenerated bone samples and the whole unilateral mandibular specimens. For the goats in the cont rol group, no operation was made and the whole unilateral mandible was taken as the test specimen. RESULTS: The compressive strength and bending stiffness of the new bone reached the normal level at 4 and 8 weeks after completion of distraction, respectively. But the shearing strength remained significantly weaker than that of the controls at 8 weeks after distraction. CONCLUSIONS: The distraction appliance can be removed and the lengthened mandible should be exposed to adaptive functional exercise at 8 weeks after completion of distraction.

Animals↗

[Mode of new bone formation following distraction osteogenesis in mandibular lengthening].

OBJECTIVE: To investigate the mode and influential factor of new bone formation following distraction osteogenesis in mandibular lengthening. METHODS: Corticotomy was performed on bilateral mandibles in twelve adult male goats. A custom-made distractor was used to lengthen the mandible at a rate of 1 mm/day for 10 days (total 10 mm elongation). Four goats were sampled respectively at 2, 4 and 8 weeks after completion of distraction. The lengthening mandibles were examined by roentgenography and histology. RESULTS: Newly formed callus was observed in the distraction gap after mandibular lengthening. The new bone exhibited intramembranous ossification generally, but cartilage islands could be found in the specimen that distractor loosed. CONCLUSION: The above findings indicate that the mode of new bone formation in mandibular lengthening following distraction osteogenesis appears to be intramembranous ossification and that endochondral ossification takes place in case distractor has loosened.

Animals↗

[Distraction osteogenesis for repair of cleft palate: an experimental study].

OBJECTIVE: To observe the repair of cleft palate (CP) bone defect under the application of distraction osteogenesis (DO); to investigate the correction mechanism in thin bone of palatal shelf and to search a possible new treatment approach for repair of CP. METHODS: 14 cats were assigned randomly to 3 groups: (1) Empty control, 2 cats unoperated with euthanasia at the end of 2 weeks; (2) Experimental control, 2 cats underwent 8 mm x 18 mm palatal osteoectomy in saggital direction as the experimental model of CP with euthanasia in 6 weeks; (3) Experimental group, 10 cats were performed the same palatal osteoectomy as group (2), the newly designed and individual manufactured intraoral distractors were fixed to teeth by brackets and to bilateral palate bone across the defect with titanium miniscrews. 4 weeks later the transport disc (TD) osteotomy were performed and gradual distraction was started after 6 days latency period at the rate of 0.4 mm x 2/day till the TD reached the opposite edge of the CP. Roentgenographic, gross meterage and ultrastructural observation were performed 2, 4, 6, 8, 12 weeks after completion of distraction. RESULTS: All the examinations revealed the TD recombination to the opposite bone edge of the CP, simultaneously, the proportional expansion of overlay mucoperiosteal flap was achieved. The bone defect were filled with de move osteogenesis. Roentgenographically, the new bone mineralization was along the DO vector, developed from the bone cut edges bilaterally to the central translucent zone until turned to thin fissure. Comparison of the plaster casts showed no change in the occlusion or dentomaxillofacial framework. CONCLUSIONS: The intraoral distractor is able to get both effective distraction and steady fixation. The examination results suggest that the CP bone and soft tissue defect could be repaired and acheived functional reconstruction under DO, which could be a new treatment approach for repair of CP.

Animals↗

[Changes in the inferior alveolar vessels and angiogenesis following mandibular lengthening with different rates of distraction].

OBJECTIVE: The purpose of this study was to study the changes in the inferior alveolar vessels and angiogenesis following mandibular lengthening with different rates of distraction. METHODS: Bilateral mandibular corticotomies were performed in 6 goats. The mandibles in 6 goats were lengthened 10.0 mm using a custom-made distractor with different rates of distraction (1.0 mm/d[n = 3] and 2.0 mm/d [n = 3]); the other 2 nondistracted mandibles served as control. The goats with distracted mandibles were killed at 2 weeks after completion of distraction. The inferior alveolar vessels with distracted calluses were harvested and processed for histologic and morphometric evaluation. RESULTS: No pathological changes in the inferior alveolar vessels were found following mandibular osteodistraction. However, the number of microvessels within distraction gap in the animals distracted at a rate of 1.0 mm/day was greater than that in the goats distracted using a rate of 2.0 mm/day, and more mature newly formed bone trabeculae was observed. CONCLUSION: There were no significant changes in the inferior alveolar vessels after mandibular lengthening with distraction osteogenesis, but rapid distraction may have adverse effects on the aniogenesis in the distraction gap.

Alveolar Process↗

[Location studies of Smad4 protein in the rat testis during postnatal development].

In order to shed light on the mechanisms of TGF-beta action in the testis,we examined the expression and function of Smad4 protein, the common-mediator Smads, which is one of intracellular signaling molecules of TGF-beta superfamily members, in rat testis during postnatal development. Whole testes were collected from SD rats aged 3 days, 7 days, 14 days and 28 days, and adult. In this study, we examined, by means of western blots, the protein expression of Smad4 during rat testicular development and its cellular localization by immunohistochemical ABC method with glucose oxidase-DAB-nickel enhancement technique. The results showed that the protein of Smad4 was present in rats from 3 days of age to adulthood, and the immunoreactivity for Smad4 was exclusively localized to the cytoplasm of Leydig cells with negative nuclei in the interstitial tissue at any time point. No expression was detected in germ cells. Therefore, our data provide evidence for the molecular mechanism of TGF-beta action in rat testes during postnatal development and spermatogenesis of rats.

Animals↗