Biomedical subjects

Ming-Hua Chen

Publications and source records attributed to Ming-Hua Chen.

15 recordsLinked to original sources

PubMed2006

[The promotive effects of N-nitrosopiperidine on the malignant transformation of the immortalized esophageal epithelium induced by human papillomavirus].

BACKGROUND: Study on the promotive effects of N-nitrosopiperidine on carcinogenesis process was performed, based on the immortalization of human fetal esophageal epithelium induced by human papillomavirus (HPV) 18E6E7 genes. METHODS: The immortalized esophageal epithelium SHEE was induced by HPV18E6E7. The cells at 17th passages were cultured in 50 ml flasks. The N-nitrosopiperidine (NPIP) 0, 2, 4, 8 mmol/L added to the cultured medium of SHEE cells for 3 weeks. The morphology, proliferation and apoptosis of the cells were studied by phase contrast microscopy and flow cytometry. Modal number of chromosomes was analyzed by standard method. Tumorigenicity of the cells was assessed by soft agar colony formation and by transplantation of cells into nude mice. Expression of HPV was detected by Western blot. RESULTS: When cells were exposed to high concentration (8 mmol/L) of NPIP, cell death was increased, leaving a few live cells. In normal cultural medium instead of NPIP proliferative status of the cells restored after 4 weeks and the cells progressed to the proliferation stage with continuous replication and atypical hyperplasia. At the end of the 8th week, the cells appeared with large colonies in soft-agar and tumor formation in transplanted nude mice. When the cells were cultured in 2, 4 mmol/L NPIP the doubling passage was delayed and without tumor formation in transplanted nude mice. Modal number of chromosomes was 61-65, in 8 mmol/L NPIP group and control group, 56-61. Expression of HPV18 appeared in experimental and control groups. CONCLUSION: NPIP promotes malignant change of the immortalized esophageal epithelial cells induced by HPV18E6E7. HPV18E6E7 synergy with NPIP will accelerate malignant transformation in esophageal epithelium.

Animals↗

PubMed2004

Intratumoral injection of arsenic to enhance antitumor efficacy in human esophageal carcinoma cell xenografts.

To enhance the therapeutic efficacy of anticancer agents and to reduce systemic side effects, it was decided to study the effect of arsenic trioxide directly on solid tumors to observe the anticancer effect of arsenic on tumors and the distribution of arsenic in tumors and other organs. Esophageal carcinoma cells were heterotransplanted in severe combined immunodeficient (SCID) mice in both laterals of the abdominal wall. When both lateral tumors had grown to approximately 10x8x5 mm3, tumor-bearing mice were used for 2 experiments. The right tumors were treated with intratumoral injection of As2O3 in 1, 5 and 10 micro g per day for 10 days sequent. The left tumors were treated with phosphate buffer solution as controls. To explore the distribution of As2O3 remaining in tumor and some organs, a single intratumoral injection of As2O3 was studied with quantitative measurement of arsenic by means of atomic absorption spectrometry. The results revealed that on the 17th day after the 1st injection As2O3-treated tumors were suppressed markedly compared to that of the contrarily lateral tumor accompanied by marked apoptosis and necrosis in tumor cells. The tumor growth inhibition (TGI) was 13.56, 62.37 and 76.92% in 1, 5 and 10 micro g group, respectively. There were no pathological changes in heart, lung, spleen, liver, kidney or brain after arsenic administration. Distribution of As2O3 revealed that As2O3 remained at higher concentration in arsenic-treated tumor tissue than in other organs. Our data suggest that intratumoral delivery of As2O3 efficiently suppresses growth of transplanted esophageal carcinoma without systemic side effects. The protocol of As2O3 intratumoral injection will be its potential clinical utility for therapy of solid tumors.

Animals↗

PubMed2004

The multistage process of carcinogenesis in human esophageal epithelial cells induced by human papillomavirus.

To investigate the multistage process of carcinogenesis, the progressive alteration of the morphology, telomerase, cytogenesis, oncogenes and tumorigenicity in the process of immortalization and malignant transformation of the human fetal esophageal epithelial cell (SHEE) was studied. The SHEE cells were immortalized by gene E6E7 of human papilloma virus (HPV) type 18 in our laboratory and continually cultivated over 100 passages, which had been malignantly transformed. Cells at the 11th, 35th, 65th and 100th passage were examined according to the following criteria: morphological changes of cell growth, contact-inhibition and anchorage-independent growth (AIG); the cell proliferative and apoptotic index; the modal number of chromosomes; c-myc, p53, bcl-2, ras; telomere length and activities of telomerase and tumorigenicity in nude mice or severe combined immunodeficient (SCID) mice. The cells of the 11th passage were well differentiated and the cells of 100th passage were relatively poorly differentiated with polymorphism, while the cells of 35th and 65th had two distinct differentiations. The proliferative indexes were 21.1%, 32.5%, 33.2%, and 40.9% and the apoptotic indexes were 3.3%, 2.7%, 3.5%, 2.7% in the 11th, 35th, 65th and 100th passage respectively. Karyotypes of four cell passages belonged to hyperdiploidy and hypotriploidy. C-myc, ras, p53 genes were low in the 10th and 35th, and high in the 65th and 100th passage, but bcl-2 was low in 4 passages. Telomere length sharply decreased from normal fetal esophagus cells until the 35th passage, but it was stably expressed in the 65th and 100th passage. The activities of telomerase were expressed in cells of the 35th, 65th and 100th passages. The efficiency of AIG varied in different passages of the SHEE cell and was absent in the 11th passage, low efficiency in the 35th passage and 65th passage, and high efficiency in the 100th passage. Transplanted cells of the 65th and 100th passage into SCID mice resulted in tumor formation, but only the 100th passage cells could grow in nude mice. All of these characteristic changes were in dynamic progressive process. These data demonstrate that carcinogenesis of esophageal epithelial cells induced by HPV is the multistage process, which goes through the initial, immortal, premalignant and malignant transformation stages. The generation of esophageal carcinoma is caused by the accumulation of cellular, genetic and molecular changes.

Agar↗

PubMed2003

Upregulated expression of Ezrin and invasive phenotype in malignantly transformed esophageal epithelial cells.

AIM: To investigate the correlation between ezrin expression and invasive phenotype formation in malignantly transformed esophageal epithelial cells. METHODS: The experimental cell line employed in the present study was originated form the progressive induction of a human embryonic esophageal epithelial cell line (SHEE) by the E6E7 genes of human papillomavirus (HPV) type 18. The cells at the 35(th) passage after induction called SHEEIMM were in a state of immortalized phase and used as the control, while that of the 85(th) passage denominated as SHEEMT represented the status of cells that were malignantly transformed. The expression changes of ezrin and its mRNA in both cell passages were respectively analyzed by RT-PCR and Western blot. Invasive phenotype was assessed in vivo by inoculating these cells into the severe combined immunodeficient (SCID) mice via subcutaneous and intraperitoneal injection, and in vitro by inoculating them on the surface of the amnion membranes, which then was determined by light microscopy and scanning electron microscopy. RESULTS: Upregulated expression of ezrin protein and its mRNA was observed in SHEEMT compared with that in SHEEIMM cells. The SHEEMT cells inoculated in SCID mice were observed forming tumor masses in both visceral organs and soft tissues in a period of 40 days with a special propensity to invading mesentery and pancreas, but did not exhibit hepatic metastases. Pathologically, these tumor cells harboring larger nucleus, nucleolus and less cytoplasm could infiltrate and destroy adjacent tissues. In the in vitro study, the inoculated SHEEMT cells could grow in cluster on the amniotic epithelial surface and intrude into the amniotic stroma. In contrast, unrestricted growth and invasiveness were not found in SHEEIMM cells in both in vivo and in vitro experiment. CONCLUSION: The upregulated ezrin expression is one of the important factors that are possibly associated with the invasive phenotype formation in malignantly transformed esophageal epithelial cells.

Animals↗

PubMed2003

Reactive oxygen species and antioxidants in apoptosis of esophageal cancer cells induced by As2O3.

To explore the relationship between the reactive oxygen species (ROS) and apoptosis in esophageal carcinoma cells (SHEE85) induced by arsenic trioxide (As2O3), we focused on changes of apoptosis, ROS, and antioxidants. Apoptosis of SHEE85 was confirmed by means of DNA fragmentation stained by Hoechst 33342, Sub-G1 cells scored by flow cytometry and ultrastructure of cells by electron microscopy. To evaluate the level of ROS, the chemiluminescent method was used for measuring the production of superoxide anion (O(-)*2). Lipid peroxide (malondialdehyde, MDA), superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) were measured respectively by the photometry method. In the cells treated with As2O3 at a concentration of 5.0 micromol/l for 2-24 h, the content of cellular O(-)*2 and MDA was increased, but SOD and GSH-Px were significantly lower in the process of apoptosis in SHEE85. As2O3 at concentration of 0.5 micromol/l did not cause cell apoptosis but promoted cell proliferation. These results suggest that As2O3 at a high dosage (5 micromol/l) causes cell apoptosis and at a low dosage (0.5 micromol/l) causes cell proliferation. The essential mechanisms of cell apoptosis induced by As2O3 may be related to the increase of ROS and decrease of anti-oxidation. ROS and antioxidants participate in the apoptotic pathway of esophageal carcinoma cells.

Antineoplastic Agents↗

PubMed2003

Ezrin, actin and cytoskeleton in apoptosis of esophageal epithelial cells induced by arsenic trioxide.

Ezrin is a key protein in membrane-cytoskeleton interaction. Expression of ezrin in actin-rich cell surfaces may play a role in the modulation of cell shape and adhesion. The aim of this study was to detect ezrin, actin and cytoskeleton and to explore their relationship in the apoptosis of esophageal epithelial cells (SHEE) induced by arsenic trioxide (As2O3). The SHEE is an immortalized human fetal esophageal epithelial cell line, and the cells were treated by administering 5, 10 and 20 micromol of As2O3. the proliferation and apoptosis of SHEE cells were examined by flow cytometry with propidium iodide staining. Ezrin expression was detected by immunocytochemistry and Western blotting. Actin filament was stained by FITC-labeled phalloidin and detected quantitatively by fluorescent microscopy. Cell morphology and microfilaments were examined by electron microscopy. The results revealed that As2O3 induced an inhibition of proliferation and the promotion of apoptosis in SHEE cells. The ezrin, actin and cytoskeleton were decreased after As2O3 treatment and the cellular morphology of apoptosis developed. Our results suggested that the morphological changes of arsenic-induced apoptosis of human esophageal epithelial cells were initiated by ezrin and actin-cytoskeletal aberrance.

Actins↗

PubMed2002

Nitric oxide and calcium ions in apoptotic esophageal carcinoma cells induced by arsenite.

AIM: To Quantitatively analyze the nitric oxide (NO) and Ca2+ in apoptosis of esophageal carcinoma cells induced by arsenic trioxide (As2O3). METHODS: The cell line SHEEC1, a malignant esophageal epithelial cell induced by HPV in synergy with TPA in our laboratory, was cultured in a serum-free medium and treated with As2O3. Before and after administration of As2O3, NO production in cultured medium was detected quantitatively using the Griess Colorimetric method. Intracellular Ca2+ was labeled by using the fluorescent dye Fluo3-AM and detected under confocal laser scanning microscope (CLSM), which was able to acquire data in real-time enabling Ca2+ dynamics of individual cells in vitro. The apoptotic cells were examined under electron microscopy. RESULTS: Intracellular concentration of Ca2+ increased from 1.00 units to 1.09-1.38 units of fluorescent intensity at As2O3 treatment and NO products subsequently released from As2O3-treated cells increased from 0.98-1.00 x10(-2)micromol x L(-1) up to 1.48-1.52 x10(-2)micromol x L(-1) and maintained in a high level continuously. Finally apoptosis of cells occurred,chromatin being agglutinated, cells shrunk,nuclei became round and mitochondria swelled. CONCLUSION: Ca2+ and NO increased with cell damage and apoptosis in cells treated by As2O3. The Ca2+ is an initial messenger to the apoptotic pathway. To investigate Ca2+ and NO will be a new direction for studying the apoptotic signaling messenger of the esophageal carcinoma cells induced by As2O3.

Antineoplastic Agents↗

PubMed2002

[Express of human papillomavirus, P53 and H-ras gene in laryngeal cancer]

OBJECTIVE: To study the abnormality of p53 and H-ras gene in laryngeal cancer cells and their relation to the infection of human papillomavirus(HPV) subtypes. METHODS: The HPV subtypes, P53 and H-ras gene were examined in 28 cases of laryngeal cancer by polymerase chain reaction (PCR), single strand conformation polymorphism(SSCP), restriction fragment length polymorphism (RFLP) and DNA sequencing. RESULTS: P53 gene mutation existed in 50%cases of laryngeal cancer and DNA sequencing showed base substitution, insertion or deletion in 4 cases; The positive rate of the H-ras oncogene mutation was only 3.57% All high risk HPV infections (HPV-16 or-18) were distinctly seen in the cancer cases, especially in cases with P53 mutation. CONCLUSION: The high risk HPV subtype infection and P53 gene abnormality may play an important role in laryngeal cancer progression and these two factors may be correlated to each other.

Journal Article↗

PubMed2002

Mitochondria, calcium and nitric oxide in the apoptotic pathway of esophageal carcinoma cells induced by As2O3.

In order to explore the early apoptotic signal messengers and to search the apoptotic pathway, the morphological and functional changes of mitochondria were examined, and nitric oxide (NO) and calcium ions (Ca2+) were measured in the course of apoptosis in esophageal carcinoma cells induced by As2O3. The esophageal carcinoma cell line SHEEC1, established by HPV in synergy with TPA in our laboratory, were cultured with serum-free medium in a culture flask, 24-well plate and small culture chambers, and added with As2O3 at 1, 3, 5 micromol/l. After 0, 2, 4, 8, 12, 24 h of drug adding the NO were measured from extracellular cultured medium and the SHEEC1 cells were collected from flasks for electron microscopic examination. Fluorescent intensity (FI) of rhodamine 123 (Rho123) labeled cells was detected using laser confocal scanning microscope (LCSM) for evaluation of mitochondrial membrane potential. Intracellular Ca2+ of cells in small culture chambers labeled with Fluo-3 dye were measured using LCSM over time. After adding As2O3, SHEEC1 cells revealed characteristic morphological and functional changes of mitochondria such as hyperplasia, swelling and disruption, accompanying decrease of transmembrane potential (FI of Rho123 decreased). The Ca2+ level increased at once after adding As2O3 and the NO concentration increased step by step till 24 h, then apoptotic morphology of cells occurred. The results suggest that by inducement of As2O3 increasing Ca2+ and NO, the apoptotic signal messengers, will initiate the mitochondria-dependent apoptotic pathway.

Antineoplastic Agents↗

PubMedDate not supplied

Early-onset listeriosis in prematurity.

Listeria monocytogenes has been recognized as a human pathogen for more than 70 years. It causes illness mainly in pregnant women, newborns, elderly, and immunocompromised persons. Although L. Monocytogenes is a relatively uncommon pathogen in neonates, it can cause considerable morbidity and mortality in this age group, especially in the early-onset form of the disease. In Taiwan, neonatal listeriosis is rarely reported. We report one case of a premature newborn with early-onset listeria sepsis and meningitis.

Female↗

PubMedDate not supplied

E6/E7 genes of human papilloma virus type 18 induced immortalization of human fetal esophageal epithelium.

To study the role played by human papilloma virus (HPV) in carcinogenesis, immortalized esophageal epithelial cells were induced by E6 and E7 genes of HPV type 18 and the biological behavior was studied. Human fetal esophageal epithelial cells were transfected with recombined HPV18E6E7AAV and were cultured and passaged in medium M199. In both the 10th passage (SHEE10) and the 31st passage (SHEE31), their proliferative rates by flow cytometry and their abilities to grow and form colonies in soft agar, or to form tumors in SCID mice were examined. The HPV18 genes of E6E7 and its expression were determined using PCR methods. Cellular telomerase activity was detected by TRAP and chromosomes were analyzed by standard method. Immortalized cell lines of esophageal epithelium induced by the HPV18E6E7 were successfully established and cultured for >100 passages over 4 years. The result of PCR showed that the E6E7 gene of HPV18 was detectable in both cell clones. Both of them were unable to grow in soft-agarose medium and failed to produce tumors in SCID mice. Flow cytometry demonstrated an average of 43% proliferation index in SHEE31, but 28% in SHEE10. Telomerase activity was clearly identified in SHEE31 but not in SHEE10. Cytogenetic analysis demonstrated progression of chromosomal abnormalities with increasing trisome. Our data indicated that genes E6/E7 of the HPV18 were capable of inducing immortalization in fetal esophageal epithelial cells. The immortal phenotype requires both activation of telomerase and genetic alterations that abrogate normal differentiation and promote cellular proliferation. This cell line can assist us to characterize the role played by HPV in carcinogenesis.

Agar↗

PubMedDate not supplied

The inhibition of growth and angiogenesis in heterotransplanted esophageal carcinoma via intratumoral injection of arsenic trioxide.

To investigate the antitumor action of arsenic trioxide (As2O3) by intratumoral injection into solid tumors, tumor growth inhibition (TGI) and angiogenesis of heterotransplanted esophageal carcinoma in mice was carried out. The cultured human esophageal carcinoma cells were inoculated into both laterals of the abdominal wall of severe combined immunodeficient (SCID) mice. When both lateral tumors had grown to about 10x8x5 mm(3), the right tumors were treated with an intratumoral injection of As2O3 in dosage of 1, 5 and 10 microg per day, respectively, for 10 days sequentially. Left tumors were treated with PBS (phosphate buffer solution) as control. The weight of transplanted tumor masses were measured and counted for TGI. The tissue of tumor, liver, kidney, heart, lung and brain was examined histopathologically and tumor tissues were examined by light- or electron-microscope. Ki-67 and CD34 were assessed by immunohistochemistry and positive nuclei of Ki-67 and microvessel density (MVD) labeled by CD34 were measured. The results revealed that on the 20th day after the first injection, As2O3-treated tumors were suppressed markedly as compared with the contrarily situated tumor, accompanied by a marked apoptosis and necrosis in tumor cells. The tissue of liver, kidney, heart, lung and brain was unaffected by As2O3. MVD in tumor tissue was decreased in the right side tumor with the significant difference in the 5 micro g and 10 micro g group (p<0.01). TGI was 5.80 (p>0.05), 58.66 (p<0.01) and 73.97% (p<0.01) in the 1, 5 and 10 micro g groups respectively, but 2.21% (p>0.05) in the control group. Conclusively, a repeated administration of As2O3 (5 and 10 microg x 10) induced an increase of tumor growth inhibition and decrease of angiogenesis in the solid tumor in tumor progressive periods. These results suggest that intra-tumoral injection of As2O3 may be investigated as a modality to treat some solid tumors.

Animals↗

PubMedDate not supplied

Kawasaki disease presenting with lymphadenopathy and gastrointestinal hemorrhage: report of one case.

We report a 5-year-old boy with Kawasaki disease (KD) initially presenting with cervical lymphadenitis and complicated by gastrointestinal hemorrhage before the use of salicylates (aspirin). A previously well and fully immunized 5-year-old Japanese boy presented with a 5-day history of neck mass and fever. Physical examination showed bilateral lymphadenopathy but not nonexudative conjunctivitis, oral cavity changes, skin rash, or extremity changes. He was admitted with a diagnosis of neck lymphadenitis and treated with parenteral antibiotics. Tarry stool was noted on the second day of admission, before the definitive diagnosis of KD was made, and cimetidine was given to treat the gastrointestinal hemorrhage. On the fourth day of admission, he fulfilled the diagnostic criteria for KD (fever for 9 days plus lymphadenopathy, lip fissure, conjunctivitis, and skin rash). He was treated with intravenous immunoglobulin (2 g/kg/day) and aspirin (80 mg/kg/day), and the fever subsided promptly. Massive gastrointestinal bleeding occurred on the ninth day of admission and was treated with whole blood transfusion, after which the vital signs were stable.

Aspirin↗

PubMedDate not supplied

Cholelithiasis in Down syndrome.

Cholelithiasis in infants, children and adolescents is rare, with a prevalence rate of less than 0.5%. The aim of this study was to determine the association between Down syndrome (DS) and cholelithiasis. We conducted a prospective, controlled study on 51 subjects (age range 11-20 years old) to assess the risk factors of cholelithiasis in children and adolescents with Down syndrome. The subjects recruited in the study consisted of 51 children with Down syndrome and 253 children in the control group. There was no statistical difference in gender and age between the DS and control groups. Gallstone disease was detected in 2 females, aged 17 and 19 years old, respectively. None had cholelithiasis in the control group. The result showed that children with DS had significantly higher prevalence of cholelithiasis (3.92%) compared with controls (0%) (p < 0.01). The overall prevalence of overweight and obesity in DS group was 83%, which was significantly higher than 20% in the control group (p < 0.01). Chromosome anomaly and overweight were two significant risk factors for cholelithiasis. Clinicians should be aware of increased risk of cholelithiasis in children with DS.

Adolescent↗

PubMedDate not supplied

Imperforate hymen presenting with chronic constipation and lumbago: report of one case.

A history of unexplained low back pain associated with chronic constipation in an adolescent girl of menarchal age or in an obviously postmenarchal girl should make one consider an imperforate hymen with hematocolpos. This is a particular important differential diagnosis in the work-up of an adolescent who denies ever having had menses or sexual activity. The case of a girl with an imperforate hymen presenting with a six-month history of chronic constipation and intermittent low back pain is described.

Adolescent↗