Expression of β3GnT8 in gastric cancer and its clinical significance / 临床与实验病理学杂志

Purpose To study the expression and the clinical significance of β3GnT8,MMP-2,PCNA in gastric cancer tissue. Methods The histological chips of paraffin specimens of gastric cancer were prepared,and immunohistochemical methods were used to detect the expression of β3GnT8,MMP-2 and PCNA in gastric cancer and adjacent tissue. Results The expression of β3GnT8 and MMP-2,PCNA in gastric cancer was higher than that in adjacent tissue (P = 0. 001) . The expression of β3GnT8 and MMP-2 was significantly positively correlated with the clinical stage (P = 0. 001) ,depth of invasion (P = 0. 011) and lymph node metastasis (P = 0. 003) of gastric cancer. The expression of β3GnT8 was significantly positively correlated with that of MMP-2 (r = 0. 703,P = 0. 001) and PCNA (r = 0. 231,P = 0. 024) . The overall survival time of the β3GnT8 positive expression group was significantly shorter than that of the negative expression group(χ2 = 3. 957,P = 0. 047) . Conclusion The expression of β3GnT8 is increased in gastric cancer. β3GnT8 can promote the invasion and metastasis of gastric cancer and is associated with poor prognosis in patients with gastric cancer.

Efficiency of an Automated Reception and Turnaround Time Management System for the Phlebotomy Room

BACKGROUND: Recent advances in laboratory information systems have largely been focused on automation. However, the phlebotomy services have not been completely automated. To address this issue, we introduced an automated reception and turnaround time (TAT) management system, for the first time in Korea, whereby the patient's information is transmitted directly to the actual phlebotomy site and the TAT for each phlebotomy step can be monitored at a glance. METHODS: The GNT5 system (Energium Co., Ltd., Korea) was installed in June 2013. The automated reception and TAT management system has been in operation since February 2014. Integration of the automated reception machine with the GNT5 allowed for direct transmission of laboratory order information to the GNT5 without involving any manual reception step. We used the mean TAT from reception to actual phlebotomy as the parameter for evaluating the efficiency of our system. RESULTS: Mean TAT decreased from 5:45 min to 2:42 min after operationalization of the system. The mean number of patients in queue decreased from 2.9 to 1.0. Further, the number of cases taking more than five minutes from reception to phlebotomy, defined as the defect rate, decreased from 20.1% to 9.7%. CONCLUSIONS: The use of automated reception and TAT management system was associated with a decrease of overall TAT and an improved workflow at the phlebotomy room.

Gluconate as suitable potential reduction supplier in Corynebacterium glutamicum: Cloning and expression of gntP and gntK in Escherichia coli

Corynebacterium glutamicum is widely used in the industrial production of amino acids. We have found that this bacterium grows exponentially on a mineral médium supplemented with gluconate. Gluconate permease and Gluconokinase are expressed in an inducible form and, 6-phosphogluconate dehydrogenase, although constituvely expressed, shows a 3-fold higher specific level in gluconate grown cells than those grown in fructose under similar conditions. Interestingly, these activities are lower than those detected in the strain Escherichia coli Ml-8, cultivated under similar conditions. Additionally, here we also confirmed that this bacterium lacks 6-phosphogluconate dehydratase activity. Thus, gluconate must be metabolized through the pentose phosphate pathway. Genes encoding gluconate transport and its phosphorylation were cloned from C. glutamicum, and expressed in suitable E. coli mutants. Sequence analysis revealed that the amino acid sequences obtained from these genes, denoted as gntP and gntK, were similar to those found in other bacteria. Analysis of both genes by RT-PCR suggested constitutive expression, in disagreement with the inducible character of their corresponding activities. The results suggest that gluconate might be a suitable source of reduction potential for improving the efficiency in cultures engaged in amino acids production. This is the first time that gluconate specific enzymatic activities are reported in C. glutamicum.

shRNAs Aiming at Glycosyltransferase Inhibit Invasive and Proliferative Ability of LoVo Cell Line in vitro / 生物化学与生物物理进展

To construct expression vectors of small hairpin RNA aimed at N-acetylglucosaminyltransferase Ⅴ(GnT-Ⅴ) gene, and to investigate effects of GnT-Ⅴ shRNA on proliferation, adhesion, migration and invasion of LoVo cell line. siRNAs were designed according to the coding sequence of GnT-Ⅴ gene, shRNA expression vectors were constructed and transfected into LoVo cell line, cell lines which stably expressed low level of GnT-Ⅴ were established by G418 screening. The mRNA and protein expression of GnT-Ⅴ were measured by semi- quantitative reverse transcription polymerase chain reaction(RT-PCR) and Western blot analysis, respectively. The effects of pGPU6/GFP/Neo GnT-Ⅴ shRNA vectors on proliferation, adhesion, migration and invasion of LoVo cell line were evaluated by CCK-8 assay, heterogenous adhesion, wound closure assay, chemotactic migration and cell invasive experiment, respectively. GnT- Ⅴ shRNA expression plasmid was constructed successfully and pGPU6/GFP/Neo GnT-Ⅴ shRNA down-regulated expression of GnT-Ⅴ dramatically in LoVo cell. Expression of LoVo GnT-Ⅴ/1564 and LoVo GnT-Ⅴ/2224 dereased by 82%, 71.5% respectively at mRNA level, and 68%, 56% respectively at protein level. The more effective interfered cell line, LoVo GnT-Ⅴ/1564, was chosen to do further experiment. CCK-8 assay showed proliferation of LoVo GnT- Ⅴ/1564 was suppressed obviously, compared to proliferation of negative control group cell (P