3D shape measurement based on structured light field imaging.

In this paper, a three-dimensional (3D) shape measurement method based on structured light field imaging is proposed, which contributes to the biomedical imaging. Generally, light field imaging is challenging to accomplish the 3D shape measurement accurately, as the slope estimation method based on radiance consistency is inaccurate. Taking into consideration the special modulation of structured light field, we utilize the phase information to substitute the phase consistency for the radiance consistency in epi-polar image (EPI) at first.

A two-step DNA barcoding approach for delimiting moth species: moths of Dongling Mountain (Beijing, China) as a case study.

DNA barcoding, based on a fragment of cytochrome c oxidase I (COI) mtDNA, is as an effective molecular tool for identification, discovery, and biodiversity assessment for most animals. However, multiple gene markers coupled with more sophisticated analytical approaches may be necessary to clarify species boundaries in cases of cryptic diversity or morphological plasticity. Using 339 moths collected from mountains surrounding Beijing, China, we tested a pipeline consisting of two steps: (1) rapid morphospecies sorting and screening of the investigated fauna with standard COI barcoding approaches; (2) additional analyses with multiple molecular markers for those specimens whose morphospecies and COI barcode grouping were incongruent.

Roles of ezrin in the growth and invasiveness of esophageal squamous carcinoma cells.

Ezrin, which crosslinks the cytoskeleton and plasma membrane, is involved in the growth and metastatic potential of cancer cells. Ezrin expression in esophageal squamous cell carcinoma (ESCC) was described recently, but its roles and the underlying mechanism(s) remain unclear. In our study, we first showed that ezrin in ESCC cell is expressed in the nucleus as well as in the cytoplasm and plasma membrane.

A novel alternative spliced variant of neutrophil gelatinase-associated lipocalin receptor in oesophageal carcinoma cells.

Recent studies suggest that NGAL (neutrophil gelatinase-associated lipocalin) is a novel iron transporter with functions distinct from that of transferrin and mediates a new iron-delivery pathway. To get a better understanding of NGAL function in oesophageal carcinoma, we analysed the expression of NGAL receptors in oesophageal carcinoma cells and identified a novel spliced variant designated NgalR-3. When expressed in a heterologous system, the protein produced from this novel spliced variant exhibits the biochemical characteristics of interaction and co-localization with NGAL protein in vivo.

Expression, purification and characterization of the Hepatitis B virus entire envelope large protein in Pichia pastoris.

The current HBsAg vaccine has performed a vital role in preventing the transmission of HBV during the past 20 years. However, a number of individuals still show no response or a low response to the vaccine. In the present study, the HBV envelope large protein gene was cloned into the eukaryotic expression vector pPIC9k and was subsequently expressed in the yeast Pichia pastoris.

[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.

Purification and application of bacterially expressed chimeric protein E1E2 of hepatitis C virus.

E1 and E2 glycoproteins are structural components of hepatitis C virus (HCV) virion. They are involved in cellular receptors interaction, neutralising antibodies elicitation, and viral morphogenesis. They are considered as major candidates for anti-HCV vaccine.

[Differential expression of STRBP8, a new candidate oncogene, in cancerization of human immortalized esophageal epithelial cells].

Background & Objective: Recently, changes in composition, structure, and function of nuclear matrix proteins (NMPs) in generation and development of tumors evoked more and more attention. Separation and identification of tumor-related NMPs is a new way to search for tumor specific biomarkers, and to study tumor pathogenesis. This study was to analyze differential expression of STRBP8, one of esophageal carcinoma specific NMPs, in cancerization of immortalized human esophageal epithelial cells.

[Expression of fascin 1 gene in the process of the immortalized esophageal carcinoma carcinogenesis].

Background & Objective: Fascin 1 is the 55kDa F-actin- binding cytoskeleton protein. Fascin 1 gene was cloned from a human teratocarcinoma. Up to now, the carcinogenesis mechanism of esophageal squamous cell carcinoma is unclear.

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.

Separation and identification of differentially expressed nuclear matrix proteins between human esophageal immortalized and carcinomatous cell lines.

Aim: To separate and identify differentially expressed nuclear matrix proteins (NMPs) between the immortalized human esophageal epithelial cell line (SHEE) and the malignantly transformed esophageal carcinoma cell line (SHEEC), and to provide new ways for finding specific markers and the pathogenesis of esophageal carcinoma.

Methods: SHEE and SHEEC cell lines were used to extract NMPs. The quality of NMPs was monitored by Western blot analysis including DNA topoisomerase IIalpha, proliferation cell nuclear antigen (PCNA) and histone.

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.

Cytogenetic and molecular genetic changes in malignant transformation of immortalized esophageal epithelial cells.

The purpose of this study was to evaluate the extent to which the expression of p53, c-myc, bcl-2, ras genes and chromosomes, along with activity of hTERT, impacts on the malignant transformation of immortalized esophageal epithelial cells. The SHEE cell line was established from an embryonic esophageal epithelial cell induced by transduction of E6E7 genes of human papillomavirus type 18 (HPV18E6E7). In cells of the 85th passage (SHEE85), the malignant transformation of SHEE was confirmed by morphology, cell proliferative index and tumor formation in SCID mice.

[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.

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).

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).