Skp2 (S-phase kinase associated protein) is substrate recognition subunit of SCFSkp2 (Cul1-Rbx1-Skp1-F boxSkp2)complex,a kind of ubiquitin ligases (E3s). Skp2 involves in regulating many functions of cell,such as cell cycle progressing,cell proliferation and differentiation,as well as apoptosis.At present many studies indicated that Skp2 is closely related to tumorigenesis and prognosis.With deeper study of Skp2 in tumor, Skp2 has the potential to be a promising drug target and biomarker of prognosis in human cancer.

Objective:The study was to investigate the effects of epidermal growth factor receptor (EGFR) antibody (cetuximab,C225) combined with cyclooxygenase 2(COX-2)inhibitor (nimesulide) on the proliferation and apoptosis of colon cancer HT-29 cells, and explore the potential molecular mechanism. Methods:C225 and nimesulide alone or in combination were incubated with HT-29 cells. MTT assay was used to measure the cell proliferation. AO/EB staining and flow cytometry were used to measure the cell apoptosis. RT-PCR was used to detect the COX-2 and non steroidal anti-inflammatory drug-activated gene (NAG-1) mRNA expression levels. The expression of Akt and phosphor-Akt protein was examined by Western blotting. Results:The inhibitory effect of nimesulide combined with C225 was significantly stronger than that of nimesulide alone [(72.8±2.3)% vs (51.8±1.8)% , P<0.05]at 48 h. Typical changes of apoptosis in HT-29 cells were observed in nimesulide combined with C225 treatment group. The apoptosis rate of combined group was significantly higher than that of single nimesulide group [(57.67±0.86)% vs(33.27±1.47)%]and single C225 group [(6.27±0.55)%,P<0.05]. C225 down-regulated the expression of COX-2 mRNA. The expression of NAG-1 mRNA was up-regulated in all treatment groups and the expression of phosphor-Akt was significantly down-regulated in combined treatment group than single nimesulide or C225 group. Conclusion:Nimesulide combined with C225 has obvious synergistic effects in inducing apoptosis. The potential mechanisms may be that EGFR signaling pathway is involved in the regulation of cycloxygenase-2 expression in HT-29 cells and finally influence the proliferation and apoptosis of HT-29 cells through the up-regulation of NAG-1 and down-regulation of phosphor-Akt expression.

Purpose: To observe the efficacy of intraperitoneal chemotherapy on advanced gastric cancer after radical treatment. Methods: 64 cases of patients with stage Ⅱ - Ⅲ treated by radical surgery were selected and randomly divided into systematic plus intraperitoneal chemotherapy group and systemic chemotherapy group. 3-year survival rate and 3-year recurrence rate were recorded. Results: The recurrence rate for the intraperitoneal chemotherapy group was 18.75% while the control was 43. 33% (P

Objective To evaluate the influence of hyperthermia on T helper subgroup (T help type1/T help type2,Th1/Th2) for patients with esophageal carcinoma. Methods 22 patients of esophageal carcinoma were randomly divided into two groups. Both group patients were treated by external radiation, the total dose was 64-66 Gy/30-33 fractions. Radiotherapy was the only therapy method in group A. Hypertbermia was used twice every week, with 8-10 fractions except for radiotherapy in group B. All patients were phlebetomized at the beginning and the end of radiotherapy for examining Th1/Th2 by flow cytometry. Results Th1 was significantly decreased in group A(t = 5.33, P < 0.01). All indexes in group B had no difference (t = - 1.41, P > 0.05). TheThl in group A was decreased much more than that in group B(t = 4.28, P < 0.05). Conclusion Hyperthermia could rectify the balance of Th1/Tb2.

Objective To explore the effect of recombinant human granulocyte colony stimulating factor (rhG-CSF) mobilization on TH17/Treg cells and its impact on suppressor of cytokine signaling-3 (SOCS3) gene expression in CD4+ T cells in donors' peripheral blood.Method Sixteen donors were injected subcutaneously with rhG-CSF 5 μg/kg every day for 5 consecutive days for peripheral blood stem cells mobilization.At the first 0,3,5 day,the mononuclear cclls (MNCs) in peripheral blood or graft and serum specimens were taken.The CD4 + T cells in MNCs were sorted using immuno-magnetic beads.The ratio of TH 17 and Treg cells in MNCs,cytokines concentrations of IL-17A,IL-21,ID23 and TGFβ1 in serum,and SODC3 gene expression in CD4+ T cells were detected by using flow cytometry,ELISA,and reverse transcription real-time quantitative PCR (RT-qPCR),respectively.Results (1)The ratio of Th17 cells (CD3+ CD8 CD17+) and Treg cells (CD4+ CD25+ Foxp3+) in MNCs in peripheral blood and graft at the first 0,3 and 5 days after mobilization was (2.69 ± 0.81) ％,(0.91 ± 0.33) ％,(0.35 ± 0.12) ％,(0.21 ± 0.05) ％,and (0.56 ± 0.24) ％,(0.72 ± 0.22％),(1.59 ± 0.54) ％,(3.38 ± 0.52) ％,respectively,showing a significant declining and increasing trend respectively (P＜0.05); (2)The cytokine concentrations in serum at the first 0,3 and 5 days after mobilization were 7.33 ± 0.89,5.78 ± 1.03 and 3.32 ± 0.84 μg/L for IL-17A; 124.56 ± 15.18,117.12 ± 14.45 and 64.94 ± 11.25 μg/L for IL-21 ; 183.52 ± 59.35,280.49 ± 69.75 and 393.62 ± 57.25μg/L for TGF-β1 (P＜0.01) ; and 45.89 ± 6.95,46.25 ± 7.44 and 47.45 ± 10.75 μg/L for IL-23,respectively.The IL-17A and IL-21 concentrations showed significant declining trend,contrarily TGF-β1 with an increasing trend,while IL-23 concentration had no change.After rhG-CSF mobilization,the SOCS3 gene expression in CD4 + T cells of peripheral blood and graft at the first 0,3,5 days was gradually increased.Conclusion rhG-CSF suppresses Th17 cells and promotes regulatory T cells generation,meanwhile decreases IL-17A and IL-21 and elevates serum TGF-β1 concentrations,and contributes to CD4 + T cells differentiation to Tregs,probably by elevating SOSC3 gene expression in CD4+ T cells.

Background and purpose: Increasing evidence has indicated that polymorphisms in the microRNA (miRNA, miR) binding site of target gene can alter the ability of miRNA and modulate the risk of cancer. We aimed to investigate the association between a miR-520a binding site single nucleotide polymorphism (SNP) rs141178472 in the PIK3CA 3’UTR and the risk of colorectal cancer in a Chinese Han population. Methods:The polymorphism rs141178472 was analyzed in a case-control study, including 386 colorectal cancer patients and 394 age-and sex-matched controls. The relationship between the polymorphism and the risk of colorectal cancer was examined by statistical methods. Results:Individuals carrying the rs141178472 CC genotype or C allele had an increased risk of developing colorectal cancer (CC vs TT, OR=1.716, 95%CI:1.084-2.716, P=0.022;C vs T, OR=1.258, 95%CI:1.021-1.551, P=0.033). Furthermore, the expression of PIK3CA was detected in the peripheral blood mononucleated cell of colorectal cancer patients, suggesting that mRNA levels of PIK3CA might be associated with SNP rs141178472. Conclusion:These ifndings provide evidence that a miR-520a binding site polymorphism rs141178472 in the PIK3CA 3’UTR may play crucial roles in the etiology of colorectal cancer.

OBJECTIVETo explore the significance of application of Dannang Recipe No. 2 (DNR2), a Chinese herbal preparation, in the perioperative period of laparoscopic cholecystectomy (LC).

METHODSThree hundred and sixty patients with LC were randomly assigned to two groups, 180 in each group. The treatment group was treated with DNR2 in the perioperative period, one dose of the recipe composed of aucklandia root 10 g, red peony root 15 g, giant knotweed rhizome 10 g, scutellaria root 10 g, honeysuckle flower 15 g, forsythia fruit 10 g, rhubarb 9 g, immature bitter orange 10 g, magnolia bark 10 g, peach kernel 10 g, red sage root 20 g and licorice root 4 g, which was boiled with water and taken one dose per day, starting from the previous night of operation. The control group was treated by antibiotic with Ceftizoxime sodium 2.0 g or Levofloxacin 200 mg via intravenous dripping once 0.5 h before operation preventively and 1-3 days after operation according to patients' condition. The indexes, including recovery time of borborygmus, gas elimination and defecation, post-operation body temperature, days needing fluid transfusion, hospitalization time, incidence of infectious complication, as well as the white blood cell counting (WBC) and serum C-reactive protein (CRP) before and after operation, were analyzed and compared between groups.

RESULTSThe recovery time of borborygmus, gas elimination and defecation were shorter in the treatment group as compared with that in the control group (10.42 +/- 4.38 h vs. 17.11 +/- 6.25 h, 15.60 +/- 5.03 h vs. 32.74 +/- 9.43 h and 38.81 +/- 9.87 h vs. 56.09 +/- 11.00 h, respectively) and all the other indexes were better in the treatment group than those in the control group, showing significant difference (P < 0.01 or P < 0.05), except the incidence of postoperative infectious complication, it was similar in the two groups.

CONCLUSIONApplication of DNR2 in perioperative stage of laparoscopic cholecystectomy can effectively promote the recovery of postoperative gastrointestinal motility and suppress the occurrence of acute inflammation.

Adult ; Aged ; Aged, 80 and over ; Cholecystectomy, Laparoscopic ; Drugs, Chinese Herbal ; administration & dosage ; Female ; Gallbladder Diseases ; complications ; surgery ; Humans ; Male ; Middle Aged ; Pain, Postoperative ; drug therapy ; Perioperative Care ; Young Adult

Astract:Objective To investigate the ultrastructural changes and HSP70 expression in liver of mice after microwave irradiation with lethal dose and to explore the application of these indexes as the basis of medical identification in microwave irradiation induced death. Methods The mice were divided into the control group and the irradiation group. Mice of the irradiation group were induced death by whole body exposure to 129 W/cm2 microwave irradiation for 30 minutes. The ultrastructure of liver was observed by transmission electron micro-scope;changes of the HSP70 mRNA and protein expression in liver were detected by reverse transcription polymerase chain reaction ( RT-PCR) and Western blotting respectively. Results Liver cytoplasm was observed dissolved with points and sheets and there were mitochondri-al crest and membrane solution in the irradiation group. And the HSP70 mRNA and protein expression level increases significantly compared with the control group with statistically significant difference (P<0. 01). Conclusion Death induced by microwave irradiation could lead to liver cytoplasm dissolution, mitochondria damage, mRNA and protein expression of HSP70 up-regulation, which may be used as important diagnostic indicators of microwave irradiation induced death.

AIMTo investigate the changes in the expression of four kinds of calcium regulatory proteins mRNA on the isolated ischemia/ reperfusion (IR) hearts.

METHODSThe rat hearts were divided into two groups: control group and IR group which received 45 min exposure to Krebs-Henseleit solution after 15 min zero-flow global ischemia. The indexes of left ventricular function, such as LVDP, +dp/dt(max), -dp/dt(max), and an arrhythmia scoring system were compared between the two groups. The messenger ribonucleic acid (mRNA) amount of sarcoplasmic reticulum (SR) Ca2+ ATPase (SERCA), phospholamban (PLB), inositol 1,4,5-trisphosphate receptor2 (IP3R2) and ryanodine receptor2 (RyR2) was measured by reverse transcription-polymerase chain reaction (RT-PCR) and normalized to the mRNA levels of beta-actin.

RESULTSIn the IR group, LVDP, +dp/dt(max) and -dp/dt(min) of the isolated hearts were depressed and the high rate of arrhythmias occurred during reperfusion. The levels of SERCA, IP3R2, RyR2 mRNA were lower in the IR isolated hearts group than those in the control group, while there was no difference in the level of phospholamban.

CONCLUSIONThese data suggest that myocardial ischemia/reperfusion can induce the depression of cardiac performance and an increased risk of arrhythmias, concomitant with the decrease in SERCA, IP3R2, RyR2 mRNA steady state levels.

Animals ; Myocardial Reperfusion Injury ; metabolism ; Myocardium ; metabolism ; RNA, Messenger ; genetics ; Rats ; Rats, Sprague-Dawley ; Sarcoplasmic Reticulum Calcium-Transporting ATPases ; genetics ; metabolism

OBJECTIVETo observe the changes of intestinal flora in diarrhea type irritable bowel syndrome with Pi-wei dampness-heat syndrome (IBS-PDS).

METHODSThe seven kinds of common intestinal bacteria in feces, including enteri bacillus, enterococci, saccharomycete, bifid bacteria, lactobacillus, bacteroides and peptococcus were studied in 21 patients suffered from IBS-PDS, and compared with those in 22 patients with IBS with deficiency of Pi syndrome (DPS) and 25 healthy subjects as control.

RESULTSAs compared with the healthy subjects, the levels of enteri bacillus and enterococci were significantly increased (P<0.01), the levels of bifid bacteria, Lactobacillus and Peptococcus were significantly decreased (P < 0.01), and saccharomycete and Bacteroides were insignificantly different in patients with PDS. As compared with patients with DPS, the levels of enteri bacillus, enterococci, bifid bacteria, Lactobacillus, Peptococcus and Bacteroidaceae were significantly increased except the level of saccharomycete.

CONCLUSIONThere may be alteration of intestinal flora in patients with IBS-PDS.

Adult ; Bifidobacterium ; isolation & purification ; Diagnosis, Differential ; Diarrhea ; etiology ; microbiology ; Enterobacteriaceae ; isolation & purification ; Female ; Humans ; Intestines ; microbiology ; Irritable Bowel Syndrome ; complications ; microbiology ; Lactobacillus ; isolation & purification ; Male ; Medicine, Chinese Traditional ; Middle Aged