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Biomedical subjects

Bing Chen

Publications and source records attributed to Bing Chen.

At least 19 recordsLinked to original sources

Using multifocal ERG responses to discriminate diabetic retinopathy.

PURPOSE: Increase the accuracy in classifying subjects with or without early diabetic retinopathy, by analyzing the multifocal electroretinogram (mfERG) responses. METHOD: The mfERG was recorded for 14 control subjects (Normal) and 26 non-insulin dependent diabetic patients, 16 of the patients had no apparent retinopathy (NDR), the other 10 had mild to moderate non-proliferative retinopathy (NPDR). The first order kernel (K1) and the first slice of second order kernel (K21) of the mfERG were summed across the field and exported as trace arrays. The feature subset was selected from the kernel arrays by criterion of inter-intra distance and sequential forward searching strategy. Based on the selected features, Fisher's linear classifiers were trained and the classification error rates were given. RESULT: With one feature selected, the error rates of the classifiers were bigger than 23% between the NDR and NPDR subjects. When six features were selected to train the classifiers, the classification error rate decreased to 6.5% between Normal and NDR, 0 between Normal and NPDR, and 9.6% between NDR and NPDR. The criterion function of inter-intra distance was calculated for each feature in the K1 and K21 arrays. According to rank of the function value, during early DR, deformations should be easier to find at the front limb of peaks N2 and P1 on the K1 trace, and at the front limb of peak N2 on the K21 trace. As a conclusion, the multifocal ERG responses have great potentials in exploring diabetic retinopathy, assisted with the pattern classification methods.

Adult↗

Expansion of canine bone marrow-derived endothelial progenitor cells and dynamic observation.

We cultured bone marrow-derived endothelial progenitor cells by a simple ex vivo expansion method and observed the expansion efficacy. Bone marrow mononuclear cells from five mongrel adult dogs were cultured with EGM-2MV medium in culture flasks coated with fibronectin. Morphology was observed with phase contrast microscopy, and a growth curve was constructed to evaluate the efficacy of expansion. Incorporation of Dil-ac-LDL was tested to evaluate the function. At different time points, immunocytochemical staining for flk-1, CD133, and factor VIII-related antigens was done and compared to staining of endothelial cells and mesenchymal stem cells and percentages of CD133, vascular endothelial growth factor receptor 2 (VEGFR-2), and the double-positive cells measured with flow cytometry to identify the quality and efficacy of expansion. Cluster-like attached cells grew to confluence at an average time of 10 days, and the mean number of cells harvested from 1 mL of bone marrow was (1.3 +/- 0.3) x 10(6). The cells presented a cobblestone-like appearance and took up Dil-ac-LDL. Immunocytochemistry showed that flk-1, CD133, and factor VIII-related antigens were positive. Flow cytometry showed that VEGFR-2 and CD133 double-positive cells augmented 242-fold at the tenth day. Ex vivo expansion can effectively proliferate endothelial progenitor cells from bone marrow; the expansion efficacy could meet the requirements for tissue engineering of blood vessels.

AC133 Antigen↗

NOTCH1 mutations in T-cell acute lymphoblastic leukemia: prognostic significance and implication in multifactorial leukemogenesis.

PURPOSE: NOTCH signaling pathway is essential in T-cell development and NOTCH1 mutations are frequently present in T-cell acute lymphoblastic leukemia (T-ALL). To gain insight into its clinical significance, NOTCH1 mutation was investigated in 77 patients with T-ALL. EXPERIMENTAL DESIGN: Detection of NOTCH1 mutation was done using reverse transcription-PCR amplification and direct sequencing, and thereby compared according to the clinical/biological data of the patients. RESULTS: Thirty-two mutations were identified in 29 patients (with dual mutations in 3 cases), involving not only the heterodimerization and proline/glutamic acid/serine/threonine domains as previously reported but also the transcription activation and ankyrin repeat domains revealed for the first time. These mutations were significantly associated with elevated WBC count at diagnosis and independently linked to short survival time. Interestingly, the statistically significant difference of survival according to NOTCH1 mutations was only observed in adult patients (>18 years) but not in pediatric patients (< or = 18 years), possibly due to the relatively good overall response of childhood T-ALL to the current chemotherapy. NOTCH1 mutations could coexist with HOX11, HOX11L2, or SIL-TAL1 expression. The negative effect of NOTCH1 mutation on prognosis was potentiated by HOX11L2 but was attenuated by HOX11. CONCLUSION: NOTCH1 mutation is an important prognostic marker in T-ALL and its predictive value could be even further increased if coevaluated with other T-cell-related regulatory genes. NOTCH pathway thus acts combinatorially with oncogenic transcriptional factors on T-ALL pathogenesis.

Adolescent↗

Phenotypical analysis of adult rat olfactory ensheathing cells on 3-D collagen scaffolds.

Olfactory ensheathing cell (OEC) transplantation is a promising or potential therapy for spinal cord injury (SCI). However, the effects of injecting OECs directly into SCI site have been limited and unsatisfied due to the complexity of SCI. To improve the outcome, proper biomaterials are thought to be helpful since these materials would allow the cells to grow three-dimensionally and guide cell migration. In this paper, we have studied the behavior of OECs in two-dimensional (2-D) condition as well as on three-dimensional (3-D) collagen scaffolds by analyzing their phenotypes such as cell proliferation, apoptosis, morphology, and gene activities of some neurotrophic factors and myelin proteins. OECs proliferation rate was increased on 3-D collagen scaffolds compared to the 2-D culture condition. OECs on 3-D collagen scaffolds also showed less apoptosis. In addition, OECs on 3-D collagen scaffolds maintained the original spindle-shape morphology and P75NTR gene activity. NGF, BDNF, and PLP were found to be upregulated in OECs cultured on 3-D collagen scaffolds by the semi-quantitative RT-PCR approach. The results suggested that 3-D collagen scaffolds provide suitable environments for the OECs to maintain their morphology as well as several important functional phenotypes and all these could be helpful for the effective treatment of SCI.

Animals↗

Hexon-chimaeric adenovirus serotype 5 vectors circumvent pre-existing anti-vector immunity.

A common viral immune evasion strategy involves mutating viral surface proteins in order to evade host neutralizing antibodies. Such immune evasion tactics have not previously been intentionally applied to the development of novel viral gene delivery vectors that overcome the critical problem of anti-vector immunity. Recombinant, replication-incompetent adenovirus serotype 5 (rAd5) vector-based vaccines for human immunodeficiency virus type 1 and other pathogens have proved highly immunogenic in preclinical studies but will probably be limited by the high prevalence of pre-existing anti-Ad5 immunity in human populations, particularly in the developing world. Here we show that rAd5 vectors can be engineered to circumvent anti-Ad5 immunity. We constructed novel chimaeric rAd5 vectors in which the seven short hypervariable regions (HVRs) on the surface of the Ad5 hexon protein were replaced with the corresponding HVRs from the rare adenovirus serotype Ad48. These HVR-chimaeric rAd5 vectors were produced at high titres and were stable through serial passages in vitro. HVR-chimaeric rAd5 vectors expressing simian immunodeficiency virus Gag proved comparably immunogenic to parental rAd5 vectors in naive mice and rhesus monkeys. In the presence of high levels of pre-existing anti-Ad5 immunity, the immunogenicity of HVR-chimaeric rAd5 vectors was not detectably suppressed, whereas the immunogenicity of parental rAd5 vectors was abrogated. These data demonstrate that functionally relevant Ad5-specific neutralizing antibodies are focused on epitopes located within the hexon HVRs. Moreover, these studies show that recombinant viral vectors can be engineered to circumvent pre-existing anti-vector immunity by removing key neutralizing epitopes on the surface of viral capsid proteins. Such chimaeric viral vectors may have important practical implications for vaccination and gene therapy.

Adenoviridae↗

Remarkable site specificity of local transposition into the Hsp70 promoter of Drosophila melanogaster.

Heat-shock genes have numerous features that ought to predispose them to insertional mutagenesis via transposition. To elucidate the evolvability of heat-shock genes via transposition, we have exploited a local transposition technique and Drosophila melanogaster strains with EPgy2 insertions near the Hsp70 gene cluster at 87A7 to produce numerous novel EPgy2 insertions into these Hsp70 genes. More than 50% of 45 independent insertions were made into two adjacent nucleotides in the proximal promoter at positions -96 and -97, and no insertions were into a coding or 3'-flanking sequence. All inserted transposons were in inverse orientation to the starting transposon. The frequent insertion into nucleotides -96 and -97 is consistent with the DNase hypersensitivity, absence of nucleosomes, flanking GAGA-factor-binding sites, and nucleotide sequence of this region. These experimental insertions recapitulated many of the phenotypes of natural transposition into Hsp70: reduced mRNA expression, less Hsp70 protein, and decreased inducible thermotolerance. The results suggest that the distinctive features of heat-shock promoters, which underlie the massive and rapid expression of heat-shock genes upon heat shock, also are a source of evolutionary variation on which natural selection can act.

Animals↗

The influence of various genotypes on the metabolic activity of NAT2 in a Chinese population.

AIM: To determine the phenotype and genotype of NAT2 in a Chinese population and study the influence of various NAT2 genotypes on the NAT2 activity. METHODS: A reverse dot blot method was used to detect the genotype of NAT2 in 120 healthy Chinese subjects. All subjects were given a single dose of 500 mg sulphadimidine (SM(2)). The plasma concentration of SM(2) and acetyl-SM(2) (AcSM(2)) 6 h after administration was determined. Molar metabolic ratio (MR) was calculated by the ratio of AcSM(2) to AcSM(2)+SM(2). RESULTS: Totals of 53 (44.2%), 47 (39.2%) and 20 (16.7%) subjects were homozygotes for wild type (wt/wt), heterozygotes for mutant (m/wt) and homozygotes for mutant (m/m), respectively. The MR of 120 subjects was 0.714+/-0.237. Twenty subjects (16.7%) were classified as poor metabolizers. All subjects in the m/m group were poor metabolizers. The MRs of the wt/wt, m/wt and m/m groups were 0.886+/-0.060, 0.719+/-0.089 and 0.246+/-0.105 (P<0.001), respectively. There was a significant difference between different NAT2 m/wt genotypes (P<0.001) and m/m genotypes (P<0.001). MR correlated well with NAT2 genotypes (r=0.947). CONCLUSION: Various NAT2 genotypes have a significant impact on the metabolic activity of NAT2 in Chinese people.

Adult↗

Effects of codon modification on human BMP2 gene expression in tobacco plants.

Bone morphogenetic protein 2 (BMP2) has great potential in therapeutic applications. We are working on generating transgenic plants as a bioreactor to produce BMP2. We have studied the effects of codon optimization on the expression of human BMP2 (hBMP2) in tobacco plants. Three modified hBMP2 genes were transformed into tobacco under the control of either cauliflower mosaic virus 35S (CaMV35S) promoter or double-CaMV35S promoter plus alfalfa mosaic virus (AMV) enhancer. The fused beta-glucuronidase (GUS) reporter gene was used to facilitate the assay of protein expression. The results indicated that codon optimization could increase the protein expression level obviously under CaMV35S promoter. However, under relatively stronger initiation condition (double-CaMV35S promoter plus AMV enhancer), only the gene with the lowest degree of codon optimization could increase the protein expression level. Our findings suggest that the action of codon optimization may be influenced by the factors of promoter strength and A+T content in tobacco plants.

Base Sequence↗

Neoplastic transformation of human diploid fibroblasts after long-term serum starvation.

Serum starvation for several days has been considered as a positive effect on the efficiency of nuclear transfer using donor cells. The effects of longer period serum starvation are not clear while similar starvation might occur in vitro maintained cells (i.e. tissue engineering products) and in vivo such as ischemia of human tissues or organs. We found human dermis fibroblasts were transformed for about 70 days caused by serum starvation (0.5% serum). The transformed cells became round and had more than one nucleolus. In 0.5% serum medium they kept almost constant growth rate as the normal fibroblasts in 10% serum medium. Abnormal karyotype including aneuploidy and structural aberrations was observed. The transformed cells had high telomerase activities, in contrast, normal fibroblasts had no detectable telomerase activities. C-myc was up-regulated while cdk2, cyclin A, p21 were down in transformed cells. Cell transplantation into SCID nude mice confirmed that the cells had the capacity of forming solid tumors. The results indicated that long-term serum starvation could lead to cell chromosomal instability and transformation.

Animals↗

[Challenges in management of cerebral ischemia due to Takayasu's arteritis].

OBJECTIVE: To explore the management of cerebral ischemia caused by Takayasu's arteritis. METHODS: One hundred and three cases treated from 1984 to 2003 were reviewed including 92 females. Seven cases underwent ascending aorta to bilateral internal carotid artery (ICA) bypass, 38 cases to the axillary artery with graft to single ICA bypass. Six cases underwent ascending aorta to axillary bypass with 3 graft to single ICA bypasses as the second stage surgery. Three cases underwent ascending aorta to right ICA bypass with 2 graft to left ICA bypasses as well as 6 subclavian to carotid bypass, PTA in 5 and stenting in 3 cases, etc. RESULTS: Twenty-seven patients with less clinical severity received conservative therapy, 9 of them had mostly temporarily improvement, 15 had slight improvement or basically no change, 1 had hemiplegia and 2 died of stroke and myocardial infarction respectively. Surgically, the short-term effective rate was 87% and operative death 7.8%. Fifty-five patients were followed up, a mean follow-up time was 48 months, and the follow-up rate was 80.9%. The excellently, good, fair, no change and death rate were 36.4%, 38.2%, 20.0%, 3.6% and 1.8% respectively. All patients with PTA or stent had an immediate good response and all recurred within 3 to 5 months. CONCLUSIONS: For treating severe cerebral ischemia caused by Takayasu's arteritis, the ascending aorta to axillary and single ICA reconstruction or the ascending aorta-bilateral axillary bypass for patients with subclavian steal syndrome is advocated; second stage operation from graft to contralateral ICA can be carried out if necessary.

Adolescent↗

The frontal-temporal nerve triangle: a new concept of locating the motor and sensory nerves in upper third of the face rhytidectomy.

BACKGROUND: How to avoid damage to the temporal branch of the facial nerve has long been a central topic of discussion. Recently, damage to the supraorbital nerve, the auriculotemporal nerve, and other branches of the trigeminal nerve divisions has attracted much attention. Focusing on frontal and temporal rhytidectomy, the authors have investigated the course and distribution of the facial nerve branches, the supraorbital nerve, the auriculotemporal nerve, and other branches of trigeminal division. In this article, they present the concept of the frontal-temporal nerve triangle; its contents, vicinity, and clinical significances are discussed. METHODS: An anatomical study was performed using 30 temporal-parietal regions of 10 fixed adult cadavers and five fresh cadavers. A step-by-step dissection from the superficial layer to the deep layer was involved; all the measurement data were analyzed, and the mean and standard deviation were calculated and expressed in centimeters. RESULTS: The frontal-temporal nerve triangle is an approximately triangular area formed by the temporal branch of the facial nerve, the supraorbital nerve, and the auriculotemporal nerve. Together with its contents and vicinal structures, it forms a complicated three-dimensional rather than two-dimensional structure. Anatomical structures closely associated with rhytidectomy are located in or near this area. CONCLUSIONS: Acting as the anatomical body surface landmark for preoperatively locating the temporal branch, the supraorbital nerve, the auriculotemporal nerve, and its related structures, the concept of the frontal-temporal nerve triangle has practical significance in designing incisions and selecting planes of dissection in upper third of the face rhytidectomy.

Facial Nerve↗

NANOGP8 is a retrogene expressed in cancers.

Nanog is a transcription factor that plays key roles in the self-renewal and maintenance of pluripotency in human embryonic stem (ES) cells. Among Nanog's 11 pseudogenes, NANOGP8 theoretically could be a retrogene, but was considered unlikely as it has not been identified in any expressed sequence tags (ESTs). In this study, we found that NANOGP8 was expressed in several cancer cell lines and in all cancer tissues tested. The complete coding sequence was cloned and the sequence is highly homologous to that of Nanog. We were also able to detect its protein expression using anti-Nanog antibody in recombinant Escherichia coli and some cancer cell lines tested. In addition, expression of NANOGP8 in NIH3T3 cells can promote cell proliferation. The expression of NANOGP8 in cancer cell lines and cancer tissues suggests that NANOGP8 may play important roles in tumorigenesis. This work not only has potential significance in stem cell and cancer research, but it also raises the possibility that some of the human pseudogenes may have regulatory functions.

Animals↗

Trans-cyclopropanation of mycolic acids on trehalose dimycolate suppresses Mycobacterium tuberculosis -induced inflammation and virulence.

Recent studies have shown that fine structural modifications of Mycobacterium tuberculosis cell envelope lipids mediate host cell immune activation during infection. One such alteration in lipid structure is cis-cyclopropane modification of the mycolic acids on trehalose dimycolate (TDM) mediated by proximal cyclopropane synthase of alpha mycolates (pcaA), a proinflammatory lipid modification during early infection. Here we examine the pathogenetic role and immunomodulatory function of mycolic acid cyclopropane stereochemistry by characterizing an M. tuberculosis cyclopropane-mycolic acid synthase 2 (cmaA2) null mutant (Delta cmaA2) that lacks trans-cyclopropanation of mycolic acids. Although titers of WT and Delta cmaA2 organisms were identical during mouse infection, Delta cmaA2 bacteria were hypervirulent while inducing larger granulomas than WT M. tuberculosis. The hypervirulence of the Delta cmaA2 strain depended on host TNF-alpha and IFN-gamma. Loss of trans-cyclopropanation enhanced M. tuberculosis-induced macrophage inflammatory responses, a phenotype that was transferable with petroleum ether extractable lipids. Finally, purified TDM lacking trans-cyclopropane rings was 5-fold more potent in stimulating macrophages. These results establish cmaA2-dependent trans-cyclopropanation of TDM as a suppressor of M. tuberculosis-induced inflammation and virulence. In addition, cyclopropane stereochemistries on mycolic acids interact directly with host cells to both positively and negatively influence host innate immune activation.

Animals↗

The molecular cytogenetic aberration analyzed by comparative genomic hybridization and its significance in diffuse large B-cell lymphoma.

OBJECTIVE: To identify genetic alterations in diffuse large B-cell lymphoma (DLBCL) and to analyse the relationship between the genetic aberrations and the clinical characteristics. METHODS: Using comparative genomic hybridization (CGH) to investigate the genomic changes in 24 cases of DLBCL and to analyse the relationship between these aberrations and clinical parameters including Ann arbor stage, systemic symptoms, chemotherapy efficacy and survival. RESULTS: Aberrations were detected in 62.5% patients of 24 cases; the most common chromosomal alterations included loss of 6q15-21 as well as gain of 18q11-ter, of which the incidences were 20.8% and 16.7%, respectively; with comparing clinical parameters between patients with normal CGH and abnormal CGH, we found that patients with abnormal CGH suffered more from stage III-IV and had higher incidence of systemic symptoms, poor chemotherapy efficacy and poor survival (P<0.05), but there was no difference observed in the incidence of extranodal involvement between two groups. CONCLUSION: The gains and/or losses of genomic DNA from DLBCL patients are the common molecular cytogenetic aberrations; loss of 6q15-21 and gain of 18q11-ter are nonrandom event to DLBCL patients; abnormal CGH is a clinical parameter reflecting malignant progressive course and poor survival to DLBCL patients.

Chromosome Aberrations↗

[Clinical research about the distribution of perineural vascular plexus of the facial nerve].

OBJECTIVE: To objectively evaluate the usefulness and the reliability of the perineural vascular plexus as a landmark for facial nerve as well as whether it will be a landmark for identification of the facial nerve in surgery for otology and neurotology by means of investigating the location of the facial nerve for prevention of iatrogenic facial palsy. METHODS: Prospective case series were designed. Three hundred and eleven consecutive patients were studied which required tympanoplasty for chronic otitis media or microsurgery for facial nerve decompression and congenitally malformation of the ear from July 2002 to July 2005. All the patients were operated by the first author. Perineural vascular plexus as a landmark for identification of the facial nerve in surgery were observed to assess the utility. RESULTS: The well recognized perineural vascular plexus were seen on the horizontal mesotympanic segment of the nerve in 95.8% of patients (298 cases), and only in 4.2% of the patients (13 cases), the vessel plexus was difficult to identify. The 95% confidence interval was from 93.6% to 98.0%. CONCLUSIONS: The vascular plexuses around or over the horizontal portion of the facial nerve provide an early and direct indicator of the location of the facial nerve. The perineural vascular plexus could be a dependable and reliable landmark for the identification the horizontal part of the facial nerve in surgery for otology and neurotology.

Adolescent↗

Expression of Nanog gene promotes NIH3T3 cell proliferation.

Cells are the functional elements in tissue engineering and regenerative medicine. A large number of cells are usually needed for these purposes. However, there are numbers of limitations for in vitro cell proliferation. Nanog is an important self-renewal determinant in embryonic stem cells. However, it remains unknown whether Nanog will influence the cell cycle and cell proliferation of mature cells. In this study, we expressed Nanog in NIH3T3 cells and showed that expression of Nanog in NIH3T3 promoted cells to enter into S phase and enhanced cell proliferation. This suggests that Nanog gene might function in a similar fashion in mature cells as in ES cells. In addition, it may provide an approach for in vitro cell expansion.

Animals↗

Distribution of alkylphenols in the Pearl River Delta and adjacent northern South China Sea, China.

The occurrence of alkylphenols (APs) was investigated in surface water and sediments from the Pearl River Delta and adjacent northern South China Sea. Most of the water samples contained detectable amounts of APs, ranging up to 0.628 microg l(-1) for nonylphenol (NP) and 0.068 microg l(-1) for octylphenol (OP). APs were found in all of the sediment samples with concentrations ranging from 59 to 7808 microg kg(-1) for NP and from 1 to 93 microg kg(-1) for OP. The Zhujiang River showed the highest concentrations of APs in both water and sediments. Significant decrease of APs concentrations going from the Zhujiang River to the Shiziyang River was observed. The Xijiang River contained concentrations of APs slightly higher in water but relatively lower in sediments than the Lingding Bay, which might be attributed to their different hydrodynamic and sedimentary characteristics. There was a decreasing trend of APs in water from the rivers to the estuary and further to the sea on the whole. In the Lingding Bay and its outer waters, concentrations of APs in sediments increased to a maximum and then decrease seaward, which was consistent with the distribution trend of the sediment organic carbon contents. Linear regression analyses showed the concentrations of APs were markedly correlated with the sediment organic carbon contents, indicating that the sediment organic carbon is an important factor controlling the levels of APs in sediments.

China↗

[Primary experimental study on the construction of tissue engineering blood vessel].

OBJECTIVE: To explore the primary experimental methods to construct tissue engineering blood vessel. METHODS: Using the collagen-chitosan to prefabricate the scaffolds with 3-dimensional structure, the proliferated human endothelial cells (ECs), smooth muscle cells (SMCs) and fibroblasts act as the seed cells. The cells were seeded to scaffolds in two-step method, and engineering tissue were matured by static culture or bioreactor culture. Extracellular matrix contents and the platelet aggregation were examined in engineering tissue, tissue engineering blood vessels were taken as patches to repair the man-made defaults on the rats aorta. RESULTS: The proliferated human ECs, SMCs and fibroblast can hold activity and act as seed cells. The prefabricated scaffolds, with excellently cell and tissue biocompatibility, can facilitated cells adherence and upgrowth, the cells quantities and extracellular matrix contents in engineered tissue are time dependent increase (P < 0.05). Platelet aggregation tests confirm the tissue engineering blood vessel have some anti-coagulability. Using the engineering tissue patch to repair the default, 6 aortas in 8 animal were patency till 10 days post-operation. CONCLUSIONS: The seeding cells can be seeded on the 3-dimensional collagen-chitosan scaffolds and matured, the tissue engineering blood vessel can be constructed primarily.

Animals↗