ObjectiveTo investigate the role of disco interacting protein 2 homolog B （DIP2B） in the proliferation， migration， and invasion processes of gastric cancer cells. MethodsBioinformatic analysis was performed using public datasets， including TCGA， GEPIA and TIMER2.0， to assess DIP2B expression and its clinical prognostic value in gastric cancer. DIP2B expression levels in gastric cancer cell lines were validated by Western blot and RT-qPCR. A DIP2B knockdown cell model was established via siRNA mediated gene silencing， and the knockdown efficiency was confirmed. Cell proliferation was assessed using real-time cellular analysis （RTCA） and colony formation assays. Cell migration and invasion were evaluated by Transwell assays， while flow cytometry was employed to detect changes in apoptosis and cell cycle distribution. ResultsDIP2B was significantly upregulated in gastric cancer cells and correlated with poor prognosis in patients （P<0.05）. The experimental results revealed that DIP2B knockdown markedly suppressed the proliferation， migration， and invasion of gastric cancer cells， while promoting apoptosis and inducing cell cycle arrest （P<0.05）. ConclusionDIP2B is highly expressed in gastric cancer and associated with unfavorable prognosis. Knockdown of DIP2B inhibits the proliferation， migration， and invasion of gastric cancer cells and promotes apoptosis， suggesting that DIP2B may serve as a potential prognostic biomarker and therapeutic target for gastric cancer.

Flavonoid 3-O-glucosyltransferase (3GT) is a key enzyme in the glucosidation of anthocyanins. To investigate the 3GT gene in rhododendron, we cloned an open reading frame (ORF) of 3GT gene (named Rh3GT) from Rhododendron hybridum Hort (Red cultivar) and then characterized this gene and the deduced protein in terms of the biochemical characteristics, expression level, and enzymatic function. The results showed that Rh3GT had a full length of 993 bp and encoded 330 amino acid residues. The deduced protein was hydrophilic, stable, weak acid, belonging to the glycosyltransferase family (GT-B type), with glutamine (Q) at position 44 in the PSPG box. The phylogenetic analysis showed that Rh3GT was most closely related to Vc3GT from Vaccinium corymbosum and Vm3GT from Vaccinium myrtillus. Rh3GT was expressed in the stems, leaves, and flowers and almost not expressed in the roots, with the highest expression level in petals during full blooming stage. Introduction of pCAMBIAL1302-Rh3GT into petals significantly up-regulated the expression level of Rh3GT and increased the total anthocyanin accumulation. Rh3GT was successfully expressed in Escherichia coli BL21 in the form of inclusion bodies with a size of about 36 kDa. The results of HPLC showed that the recombinant Rh3GT after denaturation, purification, and dilution could catalyze the synthesis of cyanidin and UDP-glucose to synthesize cyanidin 3-O-glucoside, indicating that the expressed protein had 3GT activity. This study provides basic data for further studying the molecular regulation mechanism of anthocyanin biosynthesis and theoretical support for molecular breeding of rhododendron.
Rhododendron/classification* ; Glucosyltransferases/metabolism* ; Cloning, Molecular ; Escherichia coli/metabolism* ; Recombinant Proteins/biosynthesis* ; Anthocyanins/biosynthesis* ; Phylogeny ; Plant Proteins/metabolism* ; Amino Acid Sequence

Flavanone 3-hydroxylase (F3H) is a key enzyme in the synthesis of phycocyanidins. In this experiment, the petals of red Rhododendron hybridum Hort. at different developmental stages were used as experimental materials. The R. hybridum flavanone 3-hydroxylase (RhF3H) gene was cloned using reverse transcription PCR (RT-PCR) and rapid-amplification of cDNA ends (RACE) techniques, and bioinformatics analyses were performed. Petal RhF3H gene expression at different developmental stages were analyzed by using quantitative real-time polymerase chain reaction (qRT-PCR). A pET-28a-RhF3H prokaryotic expression vector was constructed for the preparation and purification of RhF3H protein. A pCAMBIA1302-RhF3H overexpression vector was constructed for genetic transformation in Arabidopsis thaliana by Agrobacterium-mediated method. The results showed that the R. hybridum Hort. RhF3H gene is 1 245 bp long, with an open reading frame of 1 092 bp, encoding 363 amino acids. It contains a Fe²⁺ binding motif and a 2-ketoglutarate binding motif of the dioxygenase superfamily. Phylogenetic analysis showed that the R. hybridum RhF3H protein is most closely related to the Vaccinium corymbosum F3H protein. qRT-PCR analysis showed that the expression level of the red R. hybridum RhF3H gene tended to increase and then decrease in the petals at different developmental stages, with the highest expression at middle opening stage. The results of the prokaryotic expression showed that the size of the induced protein of the constructed prokaryotic expression vector pET-28a-RhF3H was about 40 kDa, which was similar to the theoretical value. Transgenic RhF3H Arabidopsis thaliana plants were successfully obtained, and PCR identification and β-glucuronidase (GUS) staining demonstrated that the RhF3H gene was integrated into the genome of A. thaliana plants. qRT-PCR, total flavonoid and anthocyanin contentanalysis showed that RhF3H was significantly higher expressed in the transgenic A. thaliana relative to that of the wild type, and its total flavonoid and anthocyanin content were significantly increased. This study provides a theoretical basis for investigating the function of RhF3H gene, as well as for studying the molecular mechanism of flower color in R. simsiib Planch.
Arabidopsis/metabolism* ; Rhododendron/metabolism* ; Amino Acid Sequence ; Anthocyanins/metabolism* ; Phylogeny ; Flavonoids/metabolism* ; Cloning, Molecular ; Gene Expression Regulation, Plant ; Plant Proteins/metabolism*

OBJECTIVETo investigate the impact of preoperative oral liquid carbohydrate on postoperative insulin resistance (IR) in gastric cancer patients undergoing elective resection, and to examine the association of IR index (homeostasis model assessment, HOMA-IR) with tumor necrosis factor-α (TNF-α).

METHODSBetween January 2013 and September 2013, 35 patients undergoing elective resection for gastric cancer were prospectively enrolled and randomized into two groups. Patients in trial group (n=18) received oral 500 ml of 10% glucose solution two hours before surgery. Patients in control group (n=17) were asked to fast for 8-12 hours before operation. About 300 mg of rectus abdominis and subcutaneous fatty tissues was removed before the closure of abdominal wall. Blood samples were collected to measure the serum concentration of TNF-α with double antibody sandwich ELISA in perioperative period (3-hour before operation, end of operation, 1-day and 3-day after operation). HOMA-IR was calculated on preoperative 3-hour and postoperative 1-day. Western blotting was used to detect protein expression of TNF-α. Correlation of HOMA-IR with TNF-α was examined.

RESULTSHOMA-IR on the first day after surgery was not different from that at 3-hour before surgery in trial group (P=0.090), which was significantly lower than that in control group (P=0.000). In trial group, serum TNF-α at the end of operation was higher than that at 3-hour before surgery, which declined rapidly on the first day after surgery and had no significant difference compared with that on the third day after surgery. In control group, serum TNF-α at the end of operation was also higher than that before surgery, which rose to the peak on the first day after surgery and was still higher than that at 3-hour before surgery. The TNF-α protein expression in muscle tissues of trial group was higher than that of control group (P=0.001), while no significant difference was observed between two groups in adipose tissues (P=0.987). Correlation analysis showed that HOMA-IR was positively correlated with TNF-α on the first day after surgery (r=0.832, P=0.000).

CONCLUSIONOral intake of liquid carbohydrate 2 hours before surgery can reduce the level of TNF-α, which is likely to improve the postoperative insulin resistance.

Carbohydrates ; Fasting ; Humans ; Insulin Resistance ; Postoperative Period ; Prospective Studies ; Stomach Neoplasms ; Tumor Necrosis Factor-alpha

Objective To investigate the effect of oral intake of 10%glucose solution before surgery on the perioperative safety and postoperative insulin resistance in patients undergoing resection for gastric cancer. Methods Between March 2012 and December 2012,36 patients undergoing elective resection for gastric cancer were enrolled and randomized into three groups. Patients in group A were given 500ml of 10%glucose solution for oral intake two hours before surgery. Patients in the group B were given distilled water instead of glucose. Patients in the group C were asked to fast for 8 hours before operation. Patient′s wellbeing scores on a visual analogue scales (VAS) were recorded. Blood samples were collected to measure. Homeostasis model assessment (HOMA) was applied to assess the status of insulin resistance (HOMA-IR) and insulin sensitivity index (ISI) according to the level of blood glucose and the serum concentration of insulin. The gastric residues before anesthesia and the time of exhaustion and defecation were also recorded. Results The gastric residues were not increased significantly after the intake of 10%glucose solution (P > 0.05). Intake of 10% glucose solution before surgery relieved thirst and hunger (P < 0.01). The time of exhaustion and defecation in three groups was not significantly different (P > 0.05). The HOMA-IR levels postoperative immediately, on the first day and the third day after surgery were lower in group A than that of group B and group C (P < 0.05), but the ISI levels were higher significantly (P < 0.01). Conclusion Intake of 10%glucose solution 2 hours before surgery for gastric cancer is be safe , which can relieve preoperative thirst and hunger, and improve postoperative insulin resistance.