Increasing glycolysis by deletion of kcs1 and arg82 improved S-adenosyl-L-methionine production in Saccharomyces cerevisiae.

Reprogramming glycolysis for directing glycolytic metabolites to a specific metabolic pathway is expected to be useful for increasing microbial production of certain metabolites, such as amino acids, lipids or considerable secondary metabolites. In this report, a strategy of increasing glycolysis by altering the metabolism of inositol pyrophosphates (IPs) for improving the production of S-adenosyl-L-methionine (SAM) for diverse pharmaceutical applications in yeast is presented. The genes associated with the metabolism of IPs, arg82, ipk1 and kcs1, were deleted, respectively, in the yeast strain Saccharomyces cerevisiae CGMCC 2842.

Glioma SOX2 expression decreased after adjuvant therapy.

Background: SOX2 is regarded as an important marker in stem cell. The change of SOX2 expression after adjuvant therapy in high grade glioma (HGG) remains unknown so far. Few patients with recurrent glioma have opportunity to undergo operation once again, so the recurrent glioma samples are scarce.

CLCA1 suppresses colorectal cancer aggressiveness via inhibition of the Wnt/beta-catenin signaling pathway.

Background: Chloride channel accessory 1 (CLCA1) belongs to the calcium-sensitive chloride conductance protein family, which is mainly expressed in the colon, small intestine and appendix. This study was conducted to investigate the functions and mechanisms of CLCA1 in colorectal cancer (CRC).

Methods: The CLCA1 protein expression level in CRC patients was evaluated by enzyme-linked immunosorbent assay (ELISA), immunohistochemistry (IHC), and western blotting analysis.

microRNA-1827 represses MDM2 to positively regulate tumor suppressor p53 and suppress tumorigenesis.

The tumor suppressor p53 plays a central role in tumor prevention. The E3 ubiquitin ligase MDM2 is the most critical negative regulator of p53, which binds to p53 and degrades p53 through ubiquitation. MDM2 itself is a transcriptional target of p53, and therefore, MDM2 forms a negative feedback loop with p53 to tightly regulate p53 levels and function.

[Proliferative and invasive effects of inhibitors of kinase 4(P15(ink4b) and P16(ink4a)/CDKN2) gene activation on RKO human colorectal cancer cells].

Objective: To explore the proliferation and invasive effects of inhibitors of kinase 4(INK4)(P15(ink4b) and P16(ink4a)/CDKN2) gene protein activation on RKO human colorectal cell in vivo and in vitro.

Methods: RKO human colorectal cell line was exposed to the specific DNA methyltransferase inhibitor 5-Aza-CdR and INK4(P15(ink4b) and P16(ink4a)/CDKN2) protein expression was detected by Western blotting. Soft agar cloning experiment and Transwell chamber assay were used to detect the proliferative and invasive ability in vitro.

Secreted protein acidic and rich in cysteines-like 1 suppresses aggressiveness and predicts better survival in colorectal cancers.

Purpose: Secreted protein acidic and rich in cysteines-like 1 (SPARCL1) is an extracellular matrix glycoprotein with malignancy-suppressing potential. The hypothesis that SPARCL1 reduces cancer invasiveness and predicts better survival in colorectal cancers (CRC) was investigated.

Experimental Design: Stable SPARCL1 transfectants, RKO-SPARCL1, and corresponding vector control were constructed and implanted into nude mice to generate a mouse xenograft model of liver metastasis.

[Effects of 5-Aza-2'-deoxycytidine on the carcinogenesis of colorectal cancer in mouse and the in vivo expression of p16/CDKN(2) mRNA].

Objective: To explore the effects and relationship of specific demethylation agent 5-Aza-2'-deoxycytidine (5-Aza-CdR) on colorectal cancer (CRC) induced by 1, 2-dimethylhydrazine (DMH) in mouse and the in vivo expression of cyclin-dependent kinases inhibitor p16/CDKN(2) mRNA.

Methods: A total of 40 male KM mice were randomized into 2 groups and CRC was induced by a 22-week injection of DMH. One group was interfered by specific DNA methyltransferase inhibitor 5-Aza-CdR.

Overexpression of HMGA2 promotes metastasis and impacts survival of colorectal cancers.

Purpose: This study aims to address the hypothesis that the high-mobility group A2 (HMGA2), an oncofetal protein, relates to survivability and serves as a prognostic biomarker for colorectal cancer (CRC).

Experimental Design: This is a retroprospective multiple center study. The HMGA2 expression level was determined by performing immunohistochemistry on surgical tissue samples of 89 CRCs from a training set and 191 CRCs from a validation set.

[Detection of chromosomal aberration in sporadic colorectal cancer with comparative genomic hybridization].

Objective: To investigate the chromosomal aberration in sporadic colorectal carcinoma and its association with clinicopathological features.

Methods: Comparative genomic hybridization(CGH) was used to screen the changes in the number of DNA sequence copies in 40 sporadic colorectal cancer patients in order to identify regions that contain genes important for the development and progression of colorectal cancer.

Results: In 40 sporadic colorectal cancer, frequent gain at 20 q, 12 q, 13 q, 7 p, 7 q and 16 q were found, while loss was also found at 18 q, 5 q, 4 q, 8 pand 17 p.

[Influence of microencapsulated ovary cells modified with maspin gene on the microvessel density and lung metastasis of breast carcinoma].

Objective: To observe the inhibition of maspin on the angiogenesis in tumor and lung metastasis of breast carcinoma and the feasibility of treatment of tumor by microencapsulated transgene cells in vivo.

Methods: Microencapsulated Chinese hamster ovarian epithelial cells (CHO) modified with maspin gene, CHO/pcDNA3.1/maspin cells, were prepared.

Expression of SNC73, a transcript of the immunoglobulin alpha-1 gene, in human epithelial carcinomas.

Aim: To investigate the expression of SNC73, a trans-cript of the immunoglobulin alpha-1 gene (IgA1-H chain), in human epithelia-derived tumor cells.

Methods: Total RNAs and cell lysates were prepared from five different human epithelial cell lines derived from lung, stomach, liver, skin, and breast, respectively. RT-PCR and immunoblot analysis of these five cell lines were done.

[Establishment of DNA oxidative damage model in colorectal crypt cells by hydrogen peroxide].

Objective: To induce DNA oxidative damage in colorectal crypt cells by hydrogen peroxide in vitro.

Methods: Hydrogen peroxide was diluted into 100, 50, 10, 5 and 1 micromol/L with RPMI 1640. Colorectal crypt cells were treated with peroxide for 10 min, 30 min, 1 h, 1.

Prognostic significance of bcl-2 and p53 expression in colorectal carcinoma.

Objective: This study was designed to detect the expression of bcl-2 and p53 proteins in colorectal carcinomas and to determine their association with the patient survival and stage of the diseases.

Methods: Immunohistochemistry method was used to detect the expression of bcl-2 and p53 proteins in 93 cases of colorectal carcinoma. The stain results were obtained by analyzing the clinic-pathological characteristics of patients.

[Development of ZM-1 tissue microarrayer].

ZM-1 tissue microarrayer designed by our group is manufactured in stainless steel and brass. It features an easier and faster preparation for tissue microarrays. By means of it, a group of biopsy needles are used to punch the donor tissue specimens respectively, and all the needles with the punched specimen cylinders are arranged into the array-board, where small holes have been digged to fit the needles.

Preparation and in vitro studies of microencapsulated cells releasing human tissue inhibitor of metalloproteinase-2.

Objective: To prepare microencapsulated cells releasing human tissue inhibitor of metalloproteinase-2 (TIMP-2), and investigate their biological characteristics in vitro.

Methods: Chinese hamster ovary (CHO) cells were stably transfected with a human TIMP-2 expression vector, encapsulated in barium alginate microcapsules and cultured in vitro. Morphological appearance of the microcapsules was observed under a light microscope.

Application of new tissue microarrayer-ZM-1 without recipient paraffin block.

The ZM-1 tissue microarrayer designed by our groups is manufactured in stainless steel and brass and contains many features that make TMA (tissue microarray) paraffin blocks construction faster and more convenient. By means of ZM-1 tissue microarrayer, biopsy needles are used to punch the donor tissue specimens respectively. All the needles with the punched specimen cylinders are arrayed into the array-board, with an array of small holes dug to fit the needles.

[Effects of microencapsulated CHO cells modified with maspin gene on the motility and adhesiveness of breast carcinoma cells Bcap37].

Objective: To investigate the effects of microencapsulated Chinese hamster ovary (CHO) cells modified with maspin gene on the motility and adhesiveness of breast carcinoma cells Bcap37 and to explore the possibility and feasibility of its clinical application in treatment of malignant tumors.

Methods: After the Bcap37 cells were co-cultured with the microencapsulated CHO cells modified with maspin gene, their motility and adhesion to vascular endothelial cells (ECV304), changes in CD44v6 and E-cadherin expression were examined.

Results: After the treatment, the motility of Bcap37 cells, their adhesion to vascular endothelial cells ECV304 and the CD44v6 expression were significantly reduced.

[Clinical analysis of 39 cases of heterotopic gastric mucosa in the upper esophagus].

Objective: To determine accurately the incidence of heterotopic gastric mucosa in the upper esophagus (HGMUE) in China, and to study the macroscopic and microscopic aspects of the lesions and to evaluate the clinical importance of HGMUE.

Methods: A prospective study was made among a total of 15,228 consecutive patients, 8,573 male and 6,655 female, aged 54 (8-95), undergoing gastroscopy. Disease histories of all patients were carefully inquired, especially those regarding possible complaints including discomfort of throat and swallowing pain and so on.

Expression of ST13 in colorectal cancer and adjacent normal tissues.

Aim: To investigate the in situ expression of suppression of tumorigenecity 13 (ST13) mRNA in both colorectal cancer and adjacent normal tissues.

Methods: Colorectal cancer cell lines SW1116, SW620 and CoLo205 were enrolled to confirm the feasibility of the in situ hybridization procedure. Seven colorectal cancer and adjacent normal tissues were included for RNA-RNA in situ hybridization.

[A study of the abnormalities of human epiderm in keloids and hypertrophic scars].

Objective: To investigate the abnormalities of human epiderm in keloids and hypertrophic scars.

Methods: Biopsies from ten untreated keloids (duration of disease 3 - 30 years) and ten hypertrophic scars (duration of disease 6 - 10 months) and five normal adult skin specimens. Total RNA was isolated from normal adult skin.

Significance of vascular endothelial growth factor expression and its correlation with inducible nitric oxide synthase in gastric cancer.

Aim: To investigate the clinical significance of the expression of VEGF165mRNA and the correlation with vascular endothelial growth factor (VEGF) protein and inducible nitric oxide synthase (iNO) in human gastric cancer.

Methods: We tested VEGF165mRNA expression in 31 cases of resected gastric cancer specimens and normal paired gastric mucosae by RT-PCR. Total RNA was extracted with TRIzol reagents, transcribed into cDNA with oligo (dT15) priming, inner controlled with beta-actin expression and agarose gel isolated after PCR.

Study on interleukin-18 gene transfer into human breast cancer cells to prevent tumorigenicity.

To study the effect of interleukin-18 gene transfection on the tumorigenesis of breast cancer cell line Bacp37, human breast cancer cell line Bcap37 were transfected with Lipofectamine and selected by G418. The biological expression of rhIL-18 was tested by RT-PCR and ELISA method; nude mice were injected with Bcap37 cell with or without the hIL-18 gene. The hIL-18 cDNA was successfully integrated into Bcap37 cell; 126.

[The study of 5-Aza-2'-deoxycytidine on transcription regulation of p16/CDKN2 gene demethylation in RKO human colorectal cell line].

Objective: To explore the transcription regulation of DNA 5'CpG island demethylation on p16/CDKN2 tumor suppressor gene and effects of growth on RKO human colorectal cancer cell line.

Methods: RKO cell line was exposed to the specific demethylating agent, 5-Aza-2'-deoxycytidine, for seventy-two hours to detect whether the silencing of p16/CDKN2 cell cycle regulatory gene could be reversed. Methylation-specific PCR (MSP), T-A cloning and sequence analysis were evaluated for methylation status.