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Biomedical subjects

M K Chen

Publications and source records attributed to M K Chen.

At least 19 recordsLinked to original sources
PubMed2004

Animal models for intestinal tissue engineering.

Although total parenteral nutrition prevents patients with short bowel syndrome from dying of starvation, having short bowel remains a severely debilitating condition. The best current treatment for inadequate absorptive surface area is through intestinal transplantation. However, this therapy is associated with significant morbidity and patients suffer from consequences of long-term immunosuppression. Additionally, the numbers of organs are limited. A new frontier in medicine is the field of tissue engineering. We will review the progress of intestinal bioengineering with a focus on the use of animal models. Investigators initially used autologous tissue as a patch to study intestinal regeneration. Subsequent studies focused on the use of absorbable biomaterials as a patch for tissue ingrowth. The most novel methodology consists of seeding a resorbable scaffold and implanting this construct to observe the regeneration of neointestine. Successful creation of esophagus, stomach, small bowel and colon has been demonstrated. Although these studies are preliminary, the results suggest that tissue-engineered intestine will become a real therapeutic option in the not too distant future for patients with inadequate intestinal tissue.

Absorbable Implants↗
PubMed2001

VEGF is upregulated in a neuroblastoma and hepatocyte coculture model.

BACKGROUND: We hypothesize that angiogenic factors are altered by the interaction between neuroblastoma cells and host tissues. MATERIALS AND METHODS: Human Chang hepatocytes and human neuroblastoma cells are cultured separately and in a noncontact, coculture system. Immunostaining for VEGF is performed on the cells. ELISA is used to detect vascular endothelial growth factor (VEGF), basic fibroblast growth factor, and interleukin-8 in the conditioned media. Human umbilical vein endothelial cells (HUVEC) are cultured with standard medium (control) and hepatocyte, neuroblastoma, and coculture conditioned media. After 48 and 72 h, cells are counted to determine proliferation. Finally, VEGF-blocking antibody is added to the HUVEC cultures with the conditioned media. RESULTS: VEGF is markedly elevated in the coculture medium compared to the media from hepatocytes or neuroblastoma grown alone [412.2 +/- 52 vs 235 +/- 35 or 74.5 +/- 28.5 (pg/10(6) cells), P < 0.05]. Other growth factors are almost undetectable in any of the media. Immunostaining for VEGF in the cocultured hepatocytes is decreased by almost 50%, but VEGF immunostaining is increased fourfold in the cocultured neuroblastoma cells. A significant increase in cell proliferation is seen at both 48 and 72 h when HUVEC are cultured with the coculture media. Cell proliferation is blocked with the addition of anti-VEGF antibody. CONCLUSION: The interaction of neuroblastoma with hepatocytes results in an increased production of VEGF. It stimulates endothelial cell proliferation and may enhance the tumor's metastatic potential in an autocrine fashion.

Cell Culture Techniques↗
PubMed2001

Small bowel tissue engineering using small intestinal submucosa as a scaffold.

BACKGROUND: Small intestinal submucosa (SIS) is an extracellular matrix used in tissue engineering studies to create de novo abdominal wall, urinary bladder, tendons, blood vessels, and dura mater. The purpose of this study is to evaluate the feasibility of using SIS as a scaffold for small bowel regeneration in an in situ xenograft model. MATERIALS AND METHODS: Twenty-three dogs had a partial defect created on the small bowel wall which was repaired with a SIS patch. Four dogs underwent small bowel resection with placement of an interposed tube of SIS. The animals were followed 2 weeks to 1 year. RESULTS: Three of the 23 dogs with SIS placed as a patch died shortly after surgery due to leakage from the site. The other 20 dogs survived up to time of elective necropsy with no evidence of intestinal dysfunction. At necropsy, the bowel circumference in the patched area had no stenosis. Histological evaluation showed the presence of a mucosal epithelial layer, varying amount of smooth muscle, sheets of collagen, and a serosal covering. Architecturally, the layers were not well organized in the submucosal region. An abundance of inflammatory cells was present in the early postoperative period but receded with time. All 4 dogs with a tubular segment of SIS interposed had significant problems. One had partial obstruction at 1 month, and 3 died in the early postoperative period due to leakage. CONCLUSIONS: This preliminary study suggests that SIS patches can be used for small bowel regeneration. Tubular segmental replacement is not feasible at this time.

Animals↗
PubMed2001

Expression and characterization of monocot rice cytosolic CuZnSOD protein in dicot Arabidopsis.

Cytosolic CuZnSOD removes deleterious superoxides from plant cells. In order to understand its function better, we sought to express a monocot CuZnSOD gene in transgenic Arabidopsis. We constructed a transgene using the CaMV 35S promoter to express a rice cytosolic CuZnSOD gene in Arabidopsis and generated over 200 transformants. A 16 kD polypeptide, the same size as the native rice CuZnSOD polypeptide, was detected in the transgenic Arabidopsis. Interestingly, two forms of rice CuZnSOD, rSODI and rSODII, having the same dimeric size, were detected in the transgenic plants. rSODII protein was relatively abundant but had low specific activity. In contrast, rSODI protein was relatively rare and had high specific activity. Inter-conversion of rSODI and rSODII could be achieved by the addition and removal of copper ions into the purified recombinant SOD and to the leaf extract of transgenic plants. Our analysis indicates that rSODI most likely corresponds to native rice CuZnSOD that has incorporated the Cu and Zn ions required for full activity, whereas the less active rSODII form may not have properly incorporated the necessary copper ions.

Arabidopsis↗
PubMed2001

Primary tumour volume delineation in nasopharyngeal carcinoma and correlation with 1997 AJCC tumour stage classification.

OBJECTIVE: To investigate the variability in primary tumour volume in nasopharyngeal carcinoma and compare it with the tumour staging of the 1997 American Joint Committee on Cancer. DESIGN: Prospective study. SETTING: Tertiary care centre. METHOD: A series of 33 newly diagnosed patients who were treated with high-dose radiotherapy participated in the study. MAIN OUTCOME MEASURES: Using computed tomographic scans, primary tumour volumes were measured using the summation-of-areas technique, and the variability in tumour volume was determined. The Mann-Whitney test was used for statistical analysis. RESULTS: A large variation in primary tumour volume was observed, especially in advanced-stage cases. CONCLUSIONS: Nasopharyngeal carcinoma shows considerable variability in primary tumour volume. Incorporation of primary tumour volume may lead to further refinement of the 1997 tumour staging system.

Adult↗
PubMed2000

Neuroblastoma and hepatocyte coculture conditioned media alter apoptosis.

BACKGROUND: Neuroblastoma is a childhood tumor that often displays unusual biological behavior. The tumor may present with widespread metastases that are unresponsive to aggressive treatment. At other times, both the metastases and the primary tumor may spontaneously regress without treatment. Apoptosis, or programmed cell death, is thought to play a role in the dichotomous behavior of neuroblastoma. We hypothesize that neuroblastoma cells will interact with host tissues to release mediators that affect apoptosis. MATERIALS AND METHODS: Human neuroblastoma cells and human Chang hepatocytes are grown in a noncontact, coculture system. After incubation for 4 days, the medium from the coculture system is collected. Neuroblastoma cells and Chang hepatocytes are then plated separately with the conditioned medium and their own standard growth medium as controls. After 4 days, these cells are harvested and cytospins made for immunostaining. Tumor necrosis factor alpha (TNF-alpha), Fas ligand, and Bcl-2, are measured with immunohistochemistry. Apoptosis is detected with the TUNEL method. Immunostaining data are interpreted with computer image analysis and reported as stain index. TUNEL data are reported as percentage apoptotic cells. All data are reported as means +/- SEM. Statistical analysis is performed and P < 0.05 considered significant. RESULTS: Chang hepatocytes grown in the coculture conditioned media have an increase in TNF-alpha and Fas ligand. The neuroblastoma cells have a significant decrease in Fas ligand. There is a significant increase in the number of apoptotic hepatocytes when they are cultured in the conditioned media. In contrast, the neuroblastoma cells grown in the coculture conditioned media show no increase in apoptosis. Finally, Bcl-2 is significantly increased in the neuroblastoma cells cultured in the conditioned media. CONCLUSIONS: Neuroblastoma cells grown in coculture conditioned media show increased expression of Bcl-2 and decreased Fas ligand levels. These changes should diminish apoptosis activity in the tumor cells. In contrast, the conditioned media induce elevated levels of proapoptotic mediators in the Chang hepatocytes. A tumor's ability to successfully metastasize may be dependent on mediators generated in the tumor-host interaction, and may not be just an independent characteristic of the tumor itself.

Apoptosis↗
PubMed2000

Floral spray transformation can efficiently generate Arabidopsis transgenic plants.

In this study, floral spray and floral dip were used to replace the vacuum step in the Agrobacterium-mediated transformation of a superoxide dismutase (SOD) gene into Arabidopsis. The transgene was constructed by using a CaMV 35S promoter to drive a rice cytosolic CuZnSOD coding sequence in Arabidopsis. The transgene construct was developed in binary vectors and mobilized into Agrobacterium. When Arabidopsis plants started to initiate flower buds, the primary inflorescence shoots were removed and then transformed by floral spray or floral dip. More than 300 transgenic plants were generated to assess the feasibility of floral spray used in the in planta transformation. The result indicates that the floral spray method of Agrobacterium can achieve rates of in planta transformation comparable to the vacuum-infiltration and floral dip methods. The floral spray method opens up the possibility of in planta transformation of plant species which are too large for dipping or vacuum infiltration.

Arabidopsis↗
PubMed2000

Free fluid on abdominal computed tomography scan after blunt trauma does not mandate exploratory laparotomy in children.

BACKGROUND/PURPOSE: The isolated finding of free intraperitoneal fluid on abdominal computed tomography (CT) scan after blunt trauma in adults is considered an indication for laparotomy by many trauma surgeons. The authors wished to determine if these guidelines are applicable to children. METHODS: A retrospective chart review was conducted. The authors included all children (< or =12 years of age) sustaining blunt abdominal trauma who were admitted to our institution between January 1, 1994 and November 1, 1998. RESULTS: There were 814 children admitted, and 437 had abdominal CT scans. Thirty-four studies showed free fluid associated with solid organ injuries, spine or pelvic fractures, or pneumoperitoneum, and were excluded. Thirty-two children had free fluid without associated injuries and formed the basis for the study. Five of these children underwent laparotomy based on the CT finding alone. The remaining 27 were observed with serial abdominal examinations and did not require surgical intervention. Only 1 of the 5 children who underwent surgery for the finding of isolated free fluid had a therapeutic laparotomy. In comparison, during the same period, 38 children underwent laparotomy after blunt injury based only on physical examination findings with a therapeutic laparotomy rate of 68%. The therapeutic laparotomy rate was significantly higher when the procedure was based solely on clinical examination as compared with the isolated finding or free fluid on the abdominal CT (26 of 38 v 1 of 5, P < .05). CONCLUSION: In contrast to adults, finding isolated free fluid on abdominal CT scans in children after blunt trauma does not dictate immediate surgical exploration.

Abdominal Injuries↗
PubMed2000

Effects of radiotherapy for nasopharyngeal carcinoma on the paranasal sinuses: study based on computed tomography scanning.

OBJECTIVE: The purpose of this study was to determine if radiotherapy for nasopharyngeal carcinoma causes mucosal disease of the paranasal sinuses. DESIGN: This study was a retrospective study. SETTING: This study was conducted at a tertiary care centre. METHOD: A series of 69 newly diagnosed patients, without pre-existing sinus disease, who were treated with high-dose radiotherapy participated. MAIN OUTCOME MEASURES: The prevalence, severity, and time course of mucosal abnormalities were analyzed, as judged by consecutive computed tomographies (CTs). RESULTS: The CT study revealed that 58.8% of the postirradiation scans had mucosal disease of the sinuses. The maxillary sinus had the highest prevalence (42.3%) without statistical significance (p = .10). The difference by McNemar test for two follow-up scans was not significant (p = .48) and by Kappa test was significant (p = .04). The relationship between the prevalence and the time course post radiotherapy revealed that it remained a high prevalence until after the 4-year follow-up scans. CONCLUSIONS: The results of this study confirm that chronic sinus disease is a common late complication of radiotherapy and it persists for years. Thus, aggressive treatment is indicated.

Adolescent↗
PubMed2000

Canal-wall-down tympanoplasty with mastoidectomy for advanced cholesteatoma.

OBJECTIVE: The objective of this study was to review 8.5 years of the senior author's experience with canal-wall-down mastoid surgery for extensive cholesteatoma with high-grade atelectasis and severely destructed ossicles. DESIGN: A retrospective review was conducted. SETTING: The setting was a tertiary care medical centre. METHODS: Available records consulted included 104 canal-wall-down mastoidectomy for advanced-stage cholesteatomas between July 1984 and December 1992. MAIN OUTCOME MEASURES: Recurrence, hearing results, and dry ear rate were analyzed. RESULTS: The recurrence rate was 4 of 104 (3.8%), and 9.6% of subjects suffered from recurrent otorrhea. Thirty-seven of 104 (35.6%) achieved the closure of air-bone gap within 20 dB. The availability of stapes suprastructure influenced the postoperative hearing level significantly (p < .001). CONCLUSION: Even in treating advanced cholesteatoma, canal-wall-down mastoidectomy provides a low recurrence rate, establishes a high dry ear rate, and preserves adequate hearing when the stapes suprastructure is available for reconstruction.

Adolescent↗
PubMed1999

Fas-mediated induction of hepatocyte apoptosis in a neuroblastoma and hepatocyte coculture model.

BACKGROUND: We have previously demonstrated an increase in hepatocyte apoptosis when they are cocultured with neuroblastoma cells. Death receptors in the tumor necrosis factor (TNF) family such as TNFR1 and Fas have been identified as regulators of apoptosis and may be responsible for the altered regulation of apoptosis seen in our coculture model. To evaluate the effects of released factors and remove the potential alterations induced by direct contact, a noncontact coculture system was used to study the interaction between hepatocytes and neuroblastoma cells. METHODS: Human Chang hepatocytes (HC) were plated onto Falcon cell culture inserts with 0.45-micrometer pores in the permeable membrane. Human neuroblastoma cells (NB-IMR-32) were seeded into wells of the Falcon companion plate. After 24 h, inserts containing HC were placed into wells containing NB cells and incubated for 4 days. This provided a coculture environment without actual cellular contact. Immunohistochemical staining for TNFalpha, Fas, and Fas ligand (Fas-L) was performed. Apoptosis was detected via the TUNEL method. Images were analyzed with ImagePro-Plus. Statistical analyses were done with significance determined at P < 0.05. RESULTS: Chang hepatocytes demonstrated a significant increase in the levels of TNF, Fas, and Fas-L when cocultured with neuroblastoma cells (P < 0.005). In addition, the cocultured hepatocytes had a 20-fold increase in the apoptotic rate (P < 0.001). Neuroblastoma cells had no demonstrable level of Fas or TNF when grown alone and in cocultures. Neuroblastoma cells that were grown alone had an elevated level of Fas-L, but this level diminished by 44% when cocultured with hepatocytes (P < 0.001). CONCLUSION: An upregulated TNF/Fas receptor-ligand system may be responsible for increased apoptosis in hepatocytes when cocultured with neuroblastoma. This upregulation may be due to release of neuroblastoma-derived Fas ligand into the media. Tumors may alter the regulation of apoptosis in surrounding tissues via the death receptors.

Apoptosis↗
PubMed1999

Analysis of the costs of surgery for Hirschsprung's disease: one-stage laparoscopic pull-through versus two-stage Duhamel procedure.

The purpose of this report is to evaluate the cost-effectiveness of a single-stage laparoscopic pull-through for Hirschsprung's disease compared to the traditional two-stage Duhamel procedure. In this series of 33 children, the length of hospitalization (2.5 +/- 3.5 vs 10.6 +/- 3.9, p < 0.01), cost (19,088 +/- 13,075 vs 34,110 +/- 19,443, p < 0.05), and complications were all significantly less with the laparoscopic assisted pull-through compared to the more traditional open approach.

Child↗
PubMed1999

Varicella arthritis diagnosed by polymerase chain reaction.

We report a 2-year-old girl who developed acute arthritis of the left knee 4 days after the onset of a typical varicella infection. She was first thought to have pyogenic arthritis caused by Staphylococcus aureus. Accordingly, oxacillin was administered upon hospitalization. On the third day after hospitalization, bacterial cultures of the synovial fluid and blood showed no growth and oxacillin was discontinued. Although a viral culture of the synovial fluid for varicella-zoster virus (VZV) was negative, varicella DNA was identified by means of polymerase chain reaction (PCR) with VZV-specific primers. The patient recovered spontaneously. To differentiate this condition from septic arthritis is important. PCR is a sensitive technique that can demonstrate the presence of VZV DNA in synovial fluid, even if viral cultures are negative.

Arthritis, Infectious↗
PubMed1999

Prospective surveillance of children with invasive Haemophilus influenzae disease in Taiwan.

During the 16 months from January 1, 1996 through April 30, 1997, forty-three cases of invasive Haemophilus influenzae disease were identified in residents younger than 14 years of age in Taiwan. H. influenzae serotyping was performed on all isolated specimens. There were 33 (76.7%) cases of type b disease; three (7.0%), non-type b, and seven (16.3%), nontypeable cases. Among these H. influenzae type b cases, there were 18 (54.5%) male patients and 15 (45.5%) female patients. With regard to age-distribution, nine (27.3%) patients were aged 2 to 5 years; nine (27.3%), between 1 to 2 years; fifteen (45.5%), younger than 1; and none were younger than 3 months old. Demographical study indicated that 13 patients (39.4%) located in northern Taiwan; 5 (15.2%), central Taiwan; 12 (36.4%), southern Taiwan; 3 (9.0%) from eastern Taiwan. Among the 33 H. influenzae type b cases, twenty-five (75.8%) patients had meningitis and 8 patients had other disease entities, i.e. pneumonia in 4 patients, bacteremia in 3, and cellulitis in 1. In terms of prognosis, three (9.1%) patients died, all of whom having meningitis or sepsis; 7 (21.2%) developed hydrocephalus; 2 (6.1%) had seizure disorder without hydrocephalus and 21 (63.6%) patients recovered completely without sequelae.

Adolescent↗
PubMed1998

The secreted hepatitis B precore antigen can modulate the immune response to the nucleocapsid: a mechanism for persistence.

The hepatitis B precore Ag (HBeAg) is a secreted nonparticulate version of the viral nucleocapsid hepatitis B core Ag (HBcAg), and its function is unknown. A proportion of HBeAg-specific Th cells evade deletion/anergy in HBeAg-transgenic (Tg) mice and mediate anti-HBe "autoantibody" (autoAb) production after in vivo activation with the appropriate Th cell peptide. This model system was used to determine how secretory HBeAg may effect deletion of Th cells in the periphery. For this purpose, HBeAg-Tg mice were bred with Fas and Fas ligand (FasL)-defective lpr/lpr and gld/gld mutant mice. Fas-FasL interactions mediate activation-induced apoptosis in the periphery. In HBeAg-Tg/+ mice, high-titrated anti-HBe autoAb was produced that was exclusively composed of the IgG1 isotype (i.e., Th2-like profile). In contrast, HBeAg-Tg/lpr and HBeAg-Tg/gld mice produced significantly less anti-HBe autoAb, and the IgG isotype patterns were broadened to include IgG2a, IgG2b and IgG3 as well as IgG1 (i.e., mixed Th1/Th2-like profile). These results suggest that HBeAg-specific Th1 cells are preferentially depleted by Fas-FasL-mediated interactions. The effect of circulating HBeAg on HBcAg-specific Th1 cells was also examined by transferring HBe/HBcAg-specific Th cells into dual HBeAg- and HBcAg-expressing Tg recipient mice. The presence of serum HBeAg ablated the expected Th1-mediated anti-HBc Ab response and shifted it toward a Th2 phenotype. These results suggest that in the context of a hepatitis B viral infection, circulating HBeAg has the potential to preferentially deplete inflammatory HBeAg- and HBcAg-specific Th1 cells that are necessary for viral clearance, thereby promoting hepatitis B virus persistence.

Adoptive Transfer↗
PubMed1998

Induction of apoptosis in a neuroblastoma and hepatocyte coculture model.

BACKGROUND: The dysregulation of apoptosis may alter the progression of tumor growth and explain the clinical dichotomy observed in children with neuroblastoma (NB). An overexpression of the bcl-2 proto-oncogene induces resistance to apoptosis and has been observed in unfavorable NB. We hypothesized that alterations in apoptosis may be a result of the interactions between NB and the tissues surrounding it. MATERIALS AND METHODS: Human Chang liver cells (HCL, 10(4) cells/cm2) were plated in two-chamber slides for 3 days. Human NB cells (10(5) cells/cm2) were added to one of the chambers and incubated for 3 more days. Control NB were plated under identical conditions in its own medium and in the HCL medium with growth curves measured. DNA fragmentation was detected via the TUNEL method (TdT-mediated nick end-labeling) and bcl-2 expression was determined by immunostaining. RESULTS: NB growth was unaltered by the change in medium. NB stained mildly positive for bcl-2 when plated alone but became markedly positive in coculture. Histologically, HCL and NB appeared healthy when plated alone, but a halo of apoptotic HCL was seen around NB in the coculture. When plated alone, both NB and HCL demonstrated minimal apoptotic activity as detected via the TUNEL method. In the coculture, a halo of HCL surrounding the NB exhibited markedly increased DNA fragmentation and this intensity diminished in cells distant from the NB. CONCLUSIONS: The regulation of apoptosis was altered in this coculture model of NB and HCL. HCL stimulated NB to overexpress bcl-2 and presumably become resistant to apoptosis. Conversely, NB induced the surrounding HCL to undergo apoptosis. The interaction between the local tissue and NB induced alterations in apoptosis in both cell types and resulted in a survival advantage for NB.

Apoptosis↗
PubMed1998

Predisposing factors of life-threatening deep neck infection: logistic regression analysis of 214 cases.

OBJECTIVE: In the era of antibiotics, most deep neck infections can be cured by conservative treatment, but some still result in life-threatening complications. In this study, we discuss whether or not there are predisposing factors of complicated deep neck infection. DESIGN: A retrospective chart review of patients presenting between 1988 and 1996 was conducted. METHOD: Among 214 deep neck infection patients, 18 cases resulted in lethal complications. We used the "dummy" variable with logistic regression as the statistical analysis method. RESULTS: Patients with an underlying disease, neck swelling, and delay time had a positive correlation that was statistically significance (p < .05). Patients who were older, male, with complete blood count/differential count-positive finding and fever had positive correlation, but this was not statistically significant (p > .05). CONCLUSION: Although complicated deep neck infection is a rapidly progressive disease with a high mortality, we can not thoroughly predict the prognosis or avoid it happening completely. But, if we can detect and pay more attention to the high-risk group of patients with use of aggressive therapy, the incidence of complicated deep neck infection may be reduced.

Abscess↗