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).

[Functions of neutrophil gelatinase-associated lipocalin in the esophageal carcinoma cell line SHEEC].

Neutrophil gelatinase-associated lipocalin (NGAL) is a novel member of the lipocalin family and may be a new human oncogene product, but function of NGAL is not clear in the cancer. It was recently found that NGAL was overexpressed in the progression of malignant transformation from human immortalized esophageal epithelial cell line SHEE to esophageal carcinoma cell line SHEEC. This indicated that cell line SHEEC was a good model for exploring functions of NGAL in the carcinogenesis.

[Cloning and identification of 5'-untranslated region (UTR) and 3'-untranslated region of neutrophil gelatinase-associated lipocalin (NGAL) gene from esophageal carcinoma cell line SHEEC].

Background & Objective: Neutrophil gelatinase-associated lipocalin (NGAL) was a novel member of the lipocalin family. The authors previously found that NGAL was overexpressed in the progress of malignant transformation from human immortalized esophageal epithelial cell line SHEE to esophageal carcinoma cell line SHEEC. However, the regulation mechanism of NGAL overexpression was not known.

[Comparative study of modulating effect of kidney tonifying recipe and spleen invigorating recipe on T-lymphocyte apoptosis in corticosterone treated rats].

Objective: To compare the modulating effect of Kidney tonifying recipe (KTR) and Spleen invigorating recipe (SIR) on T-lymphocyte apoptosis (TLA) in corticosterone treated rats (CTR).

Methods: Qualitative and quantitative analysis on activation-induced TLA was conducted by transmission electron microscope and TUNEL labelled flow cytometry detection.

Results: TLA percentage of CTR was 45.

[Comparative study on regulation patterns of compound prescriptions for kidney tonifying and for blood circulation activating on T-cell apoptosis related gene expression in aged rats].

Objective: To investigate the regulation pattern of the two compound prescriptions for Kidney tonifying, Yougui Yin and Bushen Yishou capsule, in down-regulating T-cell apoptosis gene expression in aged rats.

Methods: Expressions of T-cell apoptosis promoting and inhibiting genes, including Fas, FasL, Bcl-2, Bax, TNFR1 and TNFR2, as well as activity of cysteine proteinase in cascade connection, such as Caspase 8 and Caspase 3 were determined by TUNEL labeled flow cytometry and fluorescence real-time quantitative RT-PCR technique. The difference between old and young rats was compared, and the different regulation patterns of the two compound prescriptions for Kidney tonifying and their effects on Caspase activity were compared with those of compound prescription for blood circulation activating.

[Clinical and experimental study on retardation of immunosenescence by kidney tonifying principle].

Objective: To observe the clinical curative effect of Kidney tonifying method on retardation of immunosenescence and corresponding experimental study.

Methods: A randomized double blind placebo-controlled trial was used (RCT) on 22 pairs of aged subjects to elucidate the effect of Kidney tonifying recipe on the peripheral T-lymphocyte apoptosis and the Fas/FasL gene expression in them. In rats experimental study, the effects of two kinds of Chinese recipes (Kidney tonifying recipe and blood circulation promoting recipe) on the same parameters as in clinical study as well as on cell apoptosis and gene expression regulation in old rats were also observed.

[Automated microbiology identification and antibiotic susceptibility analysis system].

This article introduces the principles, characters and application prospect of Model 2000 Automated Microbiology Identification and Antibiotic Susceptibility Analysis System.

[Application of skin and soft tissue expansion in treatment of burn injury].

Objective: To evaluate the application of skin and soft tissue expansion in the treatment of deformity due to extensive severe burn injury and repair of severe deep electrical burned scalp and skull with fresh wound.

Methods: From 1988, 83 cases of application of skin and soft tissue expansion were reported. In those patients with deformity due to severe burn of large area and with whole nasal defect, soft tissue expander was used under the forehead skin graft and venter frontalis, followed by reconstruction of nose with the expanded vascularized skin flap and carved cartilago costalis as nasal frame.

Progressive transformation of immortalized esophageal epithelial cells.

Aim: To investigate the progressive transformation of immortal cells of human fetal esophageal epithelium induced by human papillomavirus, and to examine biological criteria of sequential passage of cells, including cellular phenotype, proliferative rate, telomerase, chromosome and tumorigenicity.

Methods: The SHEE cell series consisted of immortalized embryonic esophageal epithelium which was in malignant transformation when cultivated over sixty passages without co-carcinogens. Cells of the 10th, 31st, 60th and 85th passages were present in progressive development after being transfected with HPV.

Identification of differentially expressed proteins between human esophageal immortalized and carcinomatous cell lines by two-dimensional electrophoresis and MALDI-TOF-mass spectrometry.

Aim: To identify the differentially expressed proteins between the human immortalized esophageal epithelial cell line (SHEE) and the malignant transformed esophageal carcinoma cell line (SHEEC), and to explore new ways for studying esophageal carcinoma associated genes.

Methods: SHEE and SHEEC cell lines were used to separate differentially expressed proteins by two-dimensional electrophoresis. The silver-stained 2-D gels was scanned with EDAS290 digital camera system and analyzed with the PDQuest 6.

Immortal phenotype of the esophageal epithelial cells in the process of immortalization.

To search for potential biomarkers used to monitor the process of immortalization, we investigated the relative level of telomerase activity and other immortal phenotypes in the SHEE esophageal epithelial cell line. This human fetal esophageal epithelial cell line, induced by human papilloma virus (HPV) 18 E6E7, was continually propagated over 100 passages. Fourteenth passage cells (SHEE14) were cultured in a flask with a serum-free medium and continually cultured to the 30th passage (SHEE30).

Detection of human papillomavirus in esophageal carcinoma.

The aim of the study was to assess the prevalence of human papillomavirus (HPV) in the esophagus in the coastal region of Eastern Guangdong, Southern China, an area with a high incidence of esophageal carcinoma. Fresh surgical resection esophageal specimens were obtained from 176 esophageal carcinoma patients admitted to the Tumor Hospital of Shantou University Medical College. The samples were subjected to polymerase chain reaction (PCR) to detect HPV infection using consensus and type-specific primers for HPV type 6, 11, 16, and 18.

[Study on production of acrylamide by microbial method (II)--enzyme catalytic kinetics and de-active dynamics of nitrile hydratase].

The hydration reaction by microbial method is the crisis of the procedure of acrylamide production from acrylonitrile. This research studied the enzyme catalytic kinetics and de-active kinetics of nitrile hydratase in the type of free cell. Firstly, the effects of the concentration of cells, the temperature, pH value, the concentration of acrylonitrile and the concentration of acrylamide on the activity of nitrile hydratase was investigated.

[Repair of severe radiation-induced ulcer].

Objective: To explore a method to repair severe radiation-induced ulcer and evaluate its clinical effects.

Methods: From August 1988 to December 1998, 12 cases of severe radiation-induced ulcer were treated by muscular or musculocutaneous flap transfer, which included 1 case of pectoralis major muscular flap, 3 cases of three-lobed transverse rectus abdominis musculocutaneous flap, 2 cases of latissimus dorsi musculocutaneous flap, 1 case of tensor fascia lata musculocutaneous flap, 1 case of longitudinal rectus abdominis musculocutaneous and tensor fascia lata musculocutaneous flap, 1 case of gastrocnemius muscular flap, 1 case of gluteus major musculocutaneous flap, huge filleted musculocutaneous flap from lower limbs in 1 case and from upper limbs in 1 cases.

Results: All the flaps survived.

[Experimental study on the induced-differentiation of human esophageal cancer cells treated with dimethylsulfoxide].

The fact that treatment of leukemia (Acute Promyelocytic Leukemia) with ATRA (All-Trans Retinoic Acid) was so succeeded that it was considered as a good example for tumor therapy. In the treatment of solid tumors by means of induced differentiation, however, has not been yet so broken-through. DMSO (Dimethylsulfoxide) was a common and simple organic compound, which comprised a variety of biological activities.

[Study on production of acrylamide by microbial method (I)--Culture of bacterium cells and expression of high activity of nitrile hydratase].

The cultural conditions for the growth of Norcardia cell were studied in this paper. Controlling pH value, adding nutrient and optimizing the quantity of inducer during cultivation, the activity of nitrile hydratase reached 6567 u/mL (culture medium), which was the highest value appeared in native journals. In the farther hydratase experiments, no by-product, crylic acid, was detected.

Telomere and telomerase in the initial stage of immortalization of esophageal epithelial cell.

Aim: To search for the biomarker of cellular immortalization, the telomere length, telomerase activity and its subunits in cultured epithelial cells of human fetal esophagus in the process of immortalization.

Methods: The transgenic cell line of human fetal esophageal epithelium (SHEE) was established with E(6)E(7) genes of human papillomavirus (HPV) type 18 in our laboratory. Morphological phenotype of cultured SHEE cells from the 6th to 30th passages, was examined by phase contrast microscopy, the telomere length was assayed by Southern blot method, and the activity of telomerase was analyzed by telomeric repeat amplification protocol (TRAP).

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.

Human papillomavirus 16 E6 is associated with the nuclear matrix of esophageal carcinoma cells.

Aim: To explore the etiologic role of HPV infection in esophageal carcinoma, and the association of HPV-16 E6 with the nuclear matrix of carcinoma cells.

Methods: Two esophageal carcinoma cell lines,EC/CUHK1 and EC/CUHK2, were tested for HPV-16 E6 subgenetic fragment by polymerase chain reaction amplification of virus DNA associated nuclear matrix. RT-PCR and immunocytochemistry were also used to visualize the expression of E6 subgene in the cells.

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.

Morphological and functional changes of mitochondria in apoptotic esophageal carcinoma cells induced by arsenic trioxide.

Aim: To demonstrate that mitochondrial morphological and functional changes are an important intermediate link in the course of apoptosis in esophageal carcinoma cells induced by As2O3.

Methods: The esophageal carcinoma cell line SHEEC1, established in our laboratory, was cultured in 199 growth medium, supplemented with 100mL x L(-1) calf serum and 3 mol x L(-1)As2O3 (the same below). After 2, 4, 6, 12, 24 h of drug adding, the SHEEC1 cells were collected for light-and electron-microscopic examination.

[Vascular endothelial growth factor--fundamental research and experimental study in plastic surgery].

Objective: To review the fundamental research and the experimental study of vascular endothelial growth factor (VEGF).

Methods: The laboratory information and experimental study of VEGF were extensively reviewed.

Results: VEGF, as a mitogen specifically for endothelial cells, could strongly stimulate angiogenesis as well as vascular permeability, especially in hypoxia state.

Effects of microinjection of melatonin and its receptor antagonists into anterior hypothalamic area on blood pressure and heart rate in rats.

Aim: To examine the effects of microinjection of melatonin and its receptor antagonists into the anterior hypothalamic area (AHA) on blood pressure (BP) and heart rate (HR) in normotensive and stress-induced hypertensive rats.

Methods: Melatonin and its receptor antagonists were microinjected into the AHA, then BP, mean arterial pressure (MAP), and HR were synchronously recorded.

Results: Microinjection of melatonin produced a fall in MAP.

Estimation of the radionuclide distribution in sediment in coast area.

The study of the radionuclide distribution in sediment is a very important aspect in environmental impact of the low level radioactive liquid waste (LLW) from coastal nuclear facilities or nuclear power plant. Even now we do not know much about it. In this paper, a simple and useful method is put forward and it is used to estimate the nuclide distribution in sediment.

A comparative study of telomerase activity and malignant phenotype in multistage carcinogenesis of esophageal epithelial cells induced by human papillomavirus.

To examine certain characteristics of multistep carcinogenesis, we studied telomerase activity and malignant phenotypes in the immortal, premalignant and malignant stages of esophageal epithelial cells induced by HPV. An immortalized human fetal esophageal epithelial cell line (SHEE) was induced by E6E7 genes of human papillomavirus (HPV) type 18. Cells in the 10th passage, (SHEE10), 31st passage (SHEE31), 61st passage (SHEE61) and SHEE61A which were selected and expanded from anchorage-independent growth colonies of SHEE61, were examined as follows: cell morphology by electron-microscopy; the cell cycle by flow cytometry, telomerase activity by TRAP assay, tumorigenic detection including anchorage-independent growth by soft agar culture and tumor formation by inoculating cells into SCID and nude mice, and detection of HPV18 E6E7 oncoprotein by Western blot.