Expression of AQP1 Was Associated with Apoptosis and Survival of Patients in Gastric Adenocarcinoma.

RATIONALE AND OBJECTIVE: Recently, interest in the role of aquaporin 1 (AQP1) in human gastrointestinal carcinogenesis has developed. However, to date no studies have examined relationships between AQP1 expression and specific characteristics of gastric adenocarcinoma. METHODS: We investigated 109 specimens of primary gastric adenocarcinoma and their corresponding normal gastric mucosa using immunohistochemistry and reverse transcription-polymerase chain reaction (RT-PCR) to determine AQP1 expression. We then evaluated disease free survival (DFS) and overall survival (OS) in these patients in association with AQP1 expression. RESULTS: Both immunohistochemical and RT-PCR analyses identified increased AQP1 expression in tumors from patients with gastric adenocarcinoma (p < 0.001). The 3-year DFS and OS rates were higher in the AQP1-negative group than in the positive group (DFS: 77.2 vs. 52.8%, p < 0.001; OS: 85.1 vs. 70.7%, p < 0.001). The 5-year DFS and OS rates exhibited a similar trend (p < 0.001). Subgroup analysis of patients with early gastric adenocarcinoma (stages I and II) revealed a total 5-year OS of 90.0%, with 5-year OS being higher in the AQP1-negative group than in the positive group (95.2 vs. 84.2%). Furthermore, incidence of tumor recurrence following surgical treatment was significantly higher in the AQP1-positive group (4/19, 21.1%) compared with the negative group (0/21, 0%). CONCLUSIONS: Our study demonstrates that AQP1 plays an important role in gastric adenocarcinoma and may therefore represent a novel therapeutic target and prognostic marker in this disease.

Inhibitory Effect of Apigenin on Pharmacokinetics of Venlafaxine in vivo and in vitro.

OBJECTIVE: This study was conducted to investigate the effects of orally administered apigenin on the pharmacokinetics of venlafaxine (VEN) in rats and on the metabolism of VEN in human and rat liver microsomes in vitro. METHODS: Ten healthy male SD rats were randomly divided into 2 groups: A group (control group), B group (a single dose of 250 mg/kg apigenin). A single dose of 20 mg/kg VEN was administered orally 30 min after administration of apigenin (250 mg/kg). VEN plasma levels were measured by HPLC with fluorescence detection, and pharmacokinetic parameters were calculated by DAS 3.0 software. RESULTS: The single dose of 250 mg/kg apigenin significantly increased the AUC0-t of VEN by 40.9% (p < 0.05) and obviously increased the peak plasma concentration (Cmax) of VEN (p < 0.05). Furthermore, apigenin showed inhibitory effect on human and rat microsomes and the IC50 of apigenin was 58.37 and 25.73 µmol/l, respectively. CONCLUSIONS: Our results indicated that an intake of apigenin could increase VEN plasma levels and some of its pharmacokinetic parameters (AUC, Tmax). Thus, more attention should be paid when VEN was administrated combined with apigenin.

Cruciferous vegetable consumption and the risk of pancreatic cancer: a meta-analysis.

BACKGROUND: Previous studies regarding the association between cruciferous vegetable intake and pancreatic cancer risk have reported inconsistent results. We conducted a meta-analysis to demonstrate the potential association between them. METHODS: A systematic literature search of papers was conducted in March 2014 using PubMed, EMBASE, and Web of Science, and the references of the retrieved articles were screened. The summary odds ratios (ORs) with 95% confidence interval (CI) for the highest versus the lowest intake of cruciferous vegetables were calculated. RESULTS: Four cohort and five case-control studies were eligible for inclusion. We found a significantly decreased risk of pancreatic cancer associated with the high intake of cruciferous vegetables (OR 0.78, 95% CI 0.64-0.91). Moderate heterogeneity was detected across studies (P = 0.065). There was no evidence of significant publication bias based on Begg's funnel plot (P = 0.917) or Egger's test (P = 0.669). CONCLUSIONS: Cruciferous vegetable intake might be inversely associated with pancreatic cancer risk. Because of the limited number of studies included in this meta-analysis, further well-designed prospective studies are warranted to confirm the inverse association between cruciferous vegetable intake and risk of pancreatic cancer.

Treatment selection for gastric cancer with portal hypertension: clinical management.

BACKGROUND: Treatment for gastric cancer with portal hypertension must consider the eradication of the tumor and the change of hemodynamics in portal hypertension (PHT). Few reports have described the surgical procedures and postoperative complications of surgery for gastric cancer associated with PHT. METHODS: The clinical data of 22 patients with PHT undergoing curative surgery for gastric cancer during 5 years were retrospectively analyzed. For 12 patients classified in Child's class A, D2 lymph node (LN) dissection was performed, and 10 patients classified into Child's class B were treated with D1 LN dissection. Surgical treatment included total gastrectomy combined with pericardial devascularization, distal subtotal gastrectomy, distal subtotal gastrectomy combined with splenectomy, and distal subtotal gastrectomy combined with pericardial devascularization with posterior gastric artery and left inferior phrenic artery preserved. A liver biopsy was analyzed in all patients. RESULTS: Postoperative complications developed in 50 % (11/22 patients) and the mortality rate was 9 % (2/22). The rate of postoperative ascites in patients with Child's class A was much lower than in those with Child's class B (P < 0.05). "Operation time," "volume of hemorrhage," "platelet count," and "treatment of PHT" are all risk factors of liver function deterioration. However, there was no significant difference in liver function deterioration rate between patients with Child's class A and Child's class B (P > 0.05). The occurrence rate of complications in patients with PHT was much higher compared to those without with PHT (P < 0.05). CONCLUSIONS: Individualized selection of surgical approaches is crucial for treatment of gastric carcinoma accompanied by PHT. Surgical treatment should be based on preoperative TNM stage, liver function, and degree of PHT.

Latexin inhibits the proliferation of CD133+ miapaca-2 pancreatic cancer stem-like cells.

BACKGROUND: An increasing number of evidence suggests that pancreatic cancer contains cancer stem cells (CSCs), which may be relevant to the resistance of chemotherapy. Latexin (Lxn) is a negative regulator of stem cell proliferation and we investigate the effects of Lxn on CD133+ pancreatic cancer stem-like cells. METHODS: CD133+ miapaca-2 cells, a human pancreatic carcinoma cell line, were isolated and sorted by magnetic activated cell sorting and flow cytometry. The capacity for self-renewal, proliferation, and tumorigenicity of CD133+ miapaca-2 cells was determined by the floating spheres test and tumor xenograft assays. Protein and mRNA expression of Lxn in CD133+ and CD133- miapaca-2 cells were detected by Western blotting and qRT-PCR, respectively. After CD133+ miapaca-2 cells were treated with Lxn in serum-free medium (SFM), cell proliferation was assayed with a Cell Counting Kit 8 (CCK-8) and apoptosis was analyzed by flow cytometry. The protein and mRNA expression levels of Bcl-2, bax, and c-myc were also analyzed. RESULTS: We successfully isolated CD133+ miapaca-2 cells that exhibited the capacity for self-renewal in SFM, a proliferation potential in DMEM supplemented with FBS, and high tumorigenicity in nude mice. Lxn protein and mRNA expression levels in CD133+ miapaca-2 cells were significantly lower than those in CD133- cells. Lxn-treated CD133+ miapaca-2 cells exhibited increased apoptosis and low proliferation activity, down-regulation of Bcl-2 and c-myc expression, and up-regulation of Bax expression in a dose-dependent manner. CONCLUSIONS: Lxn induces apoptosis and inhibits the proliferation of CD133+ miapaca-2 cells. These changes are associated with down-regulation of Bcl-2 and c-myc and up-regulation of Bax.

Artesunate inhibits the growth of gastric cancer cells through the mechanism of promoting oncosis both in vitro and in vivo.

This study aims to investigate the significance and mechanism of artesunate involved in suppressing the proliferation of gastric cancer in vitro and in vivo. In the in-vitro experiments, artesunate inhibited the growth of gastric cancer cell lines (SGC-7901, BGC-823, and AGS) with concentration-dependent activity, with no significant effect on GES-1 cells. BGC-823 cells treated with artesunate showed the typical morphologic features of oncosis rather than apoptosis. Meanwhile, we observed calcium overload, downregulation of vascular endothelial growth factor expression, and upregulation of calpain-2 expression in the artesunate-treated BGC-823 cells. In addition, the in-vivo study showed that artesunate produced a dose-dependent tumor regression in nude mice. The antitumor activity of 240 mg/kg artesunate was similar to that of 10 mg/kg docetaxel. Furthermore, compared with the control group, no significant difference was observed in the body weight of artesunate-treated nude mice other than docetaxel-treated nude mice. These observations show that artesunate has concentration-dependent inhibitory activities against gastric cancer in vitro and in vivo by promoting cell oncosis through an impact of calcium, vascular endothelial growth factor, and calpain-2 expression.

[Curcumin combined FOLFOX induced cell apoptosis of gastric cancer and its mechanism research].

OBJECTIVE: To observe the effect of curcumin combined folinic acid fluorouracil oxaliplatin (FOLFOX) on the gastric adenocarcinoma cell line BGC-823 and to explore its possible mechanisms. METHODS: Cells were divided into five groups, i.e. the blank control group, the curcumin group, the FOLFOX group (0.1 mmol/L 5-FU +5 micromol/L oxaliplatin), and the curcumin combined FOLFOX group. CCK-8 was used to detect cell activity. The cell apoptosis was observed using Hoechst dyeing. Caspase-3 test kit was applied to test Caspase-3 vitality. The mRNA expressions of Bcl-2 and Bax were detected by real time fluorescent quantitative PCR. The expressions of Bcl-2 and Bax protein were determined by Western blot. RESULTS: The BGC-823 cells' proliferation could be inhibited, apoptosis induced, the Caspase-3 activity increased, expressions of Bcl-2 mRNA and Bcl-2 protein lowered, while Bax mRNA and Bax protein expressions increased in each medicated group. Besides, the efficacy of the curcumin combined FOLFOX group was superior to that of the curcumin group and the FOLFOX group, showing statistical difference (P < 0.01). CONCLUSION: Curcumin combined FOLFOX could significantly inhibit the proliferation of BGC-823 cells possibly via promoting Bax expression and Caspase-3 activity, inhibiting Bcl-2 expression, thus inducing apoptosis.

Histone acetyltransferases and deacetylases: molecular and clinical implications to gastrointestinal carcinogenesis.

Histone acetyltransferases and deacetylases are two groups of enzymes whose opposing activities govern the dynamic levels of reversible acetylation on specific lysine residues of histones and many other proteins. Gastrointestinal (GI) carcinogenesis is a major cause of morbidity and mortality worldwide. In addition to genetic and environmental factors, the role of epigenetic abnormalities such as aberrant histone acetylation has been recognized to be pivotal in regulating benign tumorigenesis and eventual malignant transformation. Here we provide an overview of histone acetylation, list the major groups of histone acetyltransferases and deacetylases, and cover in relatively more details the recent studies that suggest the links of these enzymes to GI carcinogenesis. As potential novel therapeutics for GI and other cancers, histone deacetylase inhibitors are also discussed.

[Comparison of preoperative T staging by oral contrast enhanced ultrasonography and double contrast enhanced ultrasonography in advanced gastric carcinoma].

OBJECTIVE: To compare the accuracy of preoperative T staging of gastric cancer by oral and intravenous contrast-enhanced gastric ultrasonography. METHODS: One hundred and forty three patients who had been diagnosed as gastric cancer by endoscopic biopsy and confirmed by pathology after operation were examined by oral and intravenous contrast-enhanced gastric ultrasonography, and they were divided into satisfied group and non-satisfied group according to the 2-D image quality of lesion. The results were compared with postoperative pathologic findings. RESULTS: All the patients with gastric cancer presented regional gastric wall thickening. Among them, 117 cases were clearly presented with good image quality. The remaining 26 cases were presented with vague profile, the ulcerative surface of lesion was filled with hyperechogenicity combined with rear shadow. The accuracy of oral contrast-enhanced ultrasonography in determining the T stage of gastric cancer was 74.1%. The accuracy in satisfied group and non-satisfied group was 78.6% and 53.8%, respectively. The enhancement pattern of 143 cases was showed as hyperenhancement during the arterial phase and hypoenhancement during the portal phase in DCUS. The accuracy of double contrast-enhanced ultrasongraphy in determining the T stage of gastric cancer was 86.7%, but the accuracy in satisfied group and non-satisfied group was 88.9% and 76.9%, respectively. There was a significant difference between the two methods (χ(2) = 9.031, P < 0.01). CONCLUSION: DCUS is more accurate than oral contrast-enhanced ultrasonography as a useful diagnostic method for preoperative T staging of gastric cancer.

[Effect of occlusal thickness design on the fracture resistance of endocrowns restored with lithium disilicate ceramic and zirconia].

OBJECTIVE: This study aimed to investigate the effect of occlusal thickness design on fracture resistance of endocrowns restored with lithium disilicate ceramic and zirconia. METHODS: A total of 24 artificial first mandibular molars were randomly divided into four groups with six teeth in each group as follows: group lithium disilicate ceramic-2 mm (lithium disilicate ceramic, with an occlusal thickness of 2 mm and a retainer length of 4 mm); group lithium disilicate ceramic-4 mm (lithium disilicate ceramic, with an occlusal thickness of 4 mm and a retainer length of 2 mm); group zirconia-2 mm (zirconia, with an occlusal thickness of 2 mm and a retainer length of 4 mm); and group zirconia-4 mm (zirconia, with an occlusal thickness of 4 mm and a retainer length of 2 mm). After adhesive cementation (RelyX Ultimate Clicker), all specimens were subjected to thermocycling (10 000 cycles). The specimens were subjected to fracture resistance testing at a 135° angle to the teeth at a crosshead speed of 0.5 mm·min⁻¹ in a universal testing machine. Data were analyzed with ANOVA and Tukey's HSD test by SPSS 15.0. The failure modes were classified. RESULTS: The fracture resistances of groups lithium disilicate ceramic-2 mm, lithium disilicate ceramic-4 mm, zirconia-2 mm, and zirconia-4 mm were (890.54±83.41), (2 320.87±728.57), (2 258.05±557.66), and (3 847.70±495.99) N respectively. Group zirconia-4 mm had the highest fracture resistance, whereas group lithium disilicate ceramic-2 mm had the lowest. CONCLUSIONS: The fracture resistance of molar endocrown with zirconia is higher than that with lithium disilicate ceramic. Increasing the occlusal thickness can improve the fracture resistance but increase the risk of fracture of abutment.

[Diagnostic value of double contrast-enhanced ultrasonography in preoperative staging of gastric cancer].

OBJECTIVE: To evaluate the clinical value of double contrast-enhanced ultrasonography using oral and intravenous contrast agents in preoperative staging of gastric cancer. METHODS: Sixty-two patients with biopsy-proven gastric cancer were enrolled into this study, and were examined by double contrast-enhanced gastric ultrasonography preoperatively. The results were compared with postoperative pathologic findings. RESULTS: The accuracy of oral contrast-enhanced gastric ultrasonography and double contrast-enhanced ultrasonography in determining the T stage of gastric cancer was 72.9% (T1: 66.7%, T2: 60.0%, T3: 76.9%, T4: 71.4%) and 88.1% (T1: 66.7%, T2: 80.0%, T3: 89.7%, T4: 100%), respectively, with a statistically significant difference between the two methods (P = 0.036). The sensitivity, specificity, accuracy and Youden index of oral contrast-enhanced gastric ultrasonography and double contrast-enhanced ultrasonography in assessment of lymph node metastasis were 74.5%, 66.7%, 72.9%, and 0.41 versus 89.4%, 75.0%, 86.4%, 0.76, respectively. No significant difference in the accuracy of assessment for lymph node metastasis was observed (P > 0.05). CONCLUSION: Double contrast-enhanced ultrasonography is useful for preoperative staging of gastric cancer, especially for T staging.

[Effects of universal adhesives and resin cement on the shear bond strength of zirconia].

OBJECTIVE: To study the effects of universal adhesives and resin cement on the shear bond strength and durability of zirconia ceramics. METHODS: Zirconia ceramics were sintered into 20 mm×10 mm×10 mm and 10 mm×10 mm×10 mm specimens. The experiment was divided into 12 groups. The two types of specimens were bonded using two variants of resin cement (RelyX Ultimate and Clearfil SAC self-adhesive resin cement), universal adhesives (non-adhesive, Scotchbond uni-versal adhesive, and Clearfil SE One adhesive), and storage conditions (water bath and water bath-thermal cycling). The shear bond strengths were tested, and the fracture morphologies were analyzed. RESULTS: The cement (F=8.41, P<0.01) and adhesive (F=30.34, P<0.01) exerted a significant effect on the shear bond strength of zirconia, whereas storage condition showed no significant effect on this property (F=1.83, P=0.18). The lowest shear bond strength (14.02 MPa±6.86 MPa) was exhibited by the group treated with RelyX Ultimate resin cement, non-adhesive, and water bath-thermal cycling, whereas the highest shear bond strength (54.12 MPa±8.37 MPa) was displayed by the group treated with RelyX Ultimate resin cement, Scotchbond universal adhesive, and water bath-thermal cycling. CONCLUSIONS: Universal adhesives can improve the durability of the bonding of resin cement to zirconia. If non-self-adhesive resin cement is used without a universal adhe-sive, the durability of the bond will be greatly reduced.

[The changes in insulin sensitivity under stress and its clinical significance in the critically ill patients].

OBJECTIVE: To observe the changes in blood glucose, serum insulin level and insulin resistance of critically ill patients during the acute stage and explore the relationship between the changes and their prognosis. METHODS: Fifty-four patients in intensive care unit (ICU) were enrolled. All the patients were assessed with acute physiology and chronic health evaluation II (APACHE II) scores in 24 hours after admission. Fasting blood glucose (FBG) and fasting insulin (FINS) were determined within 24 hours or the next morning, and then insulin resistance was evaluated by insulin sensitivity index (ISI). According to the APACHE II scores, the severity and prognosis, the patients were stratified into groups respectively. The difference in APACHE II scores, FBG, FINS and ISI among groups were analyzed. Furthermore, the correlation between APACHE II score and FBG, FINS, ISI were studied respectively. RESULTS: Based on APACHE II scores, the patients were divided into group A (APACHE II scoresor=21). The levels of FBG and FINS were found to be elevated with the increase in APACHE II score, and in contrast ISI greatly reduced (P<0.05 or P<0.01). APACHE II scores, FBG and FINS in multiple organ dysfunctions group were significantly higher than in single organ dysfunction group, and ISI was found to be significantly decreased (P<0.05 or P<0.01). Additionally, APACHE II scores and FBG in non-survivor group were obviously higher than those in survivor group, while ISI was markedly lower (all P<0.01), but no difference in FINS levels was found between the two groups. In the 54 critically ill patients, correlation study showed that significant positive correlation was found between FBG and APACHE II score (r=0.816 5, P<0.01, regression equation : y=0.573x+3.072) and significant negative correlation between ISI and APACHE II score (r=-0.703 9, P<0.01, regression equation : y=-0.107x-3.598), but no linear correlation was found between APACHE II score and FINS (r=0.283 0, P>0.05). CONCLUSION: The study suggests that FBG and ISI could be used to evaluate the severity and prognosis. Hyperinsulinemia is found in some critically ill patients, but FINS is not helpful for assessing the severity and outcome.

Hidden Blood Loss and Its Influential Factors After Laparoscopy-Assisted Gastrectomy for Gastric Cancer.

INTRODUCTION: Laparoscopy-assisted gastrectomy (LAG) is a minimally invasive procedure for the treatment of gastric cancer. It is generally thought that a minimally invasive technique results in less visible blood loss during the surgery. Nevertheless, a meaningful perioperative hidden blood loss (HBL) is often ignored. In this study, we investigated the amount of HBL and the influential factors after LAG for gastric cancer. METHODS: A retrospectively analyzed clinical data of 62 consecutive patients who underwent laparoscopy-assisted total or distal gastrectomy at our center from May 2016 to May 2017. The HBL was calculated according to Gross's and Nadler's formula. The data of patient gender, age, height, weight, body mass index, preoperative and postoperative hematocrit, postoperative drainage, albumin loss, diabetes mellitus, and hypertension were analyzed by multivariate linear regression analysis. The type of surgical reconstruction was analyzed by one-way analysis of variance. The difference between the preoperative blood pressure and postoperative blood pressure was measured by paired sample t-test and boxplot. RESULTS: The HBL was 322.2 ± 195.9 mL (64.3% ± 14.1% in total blood loss [TBL]), the TBL was 475.6 ± 222.8 mL, and the hemoglobin (HB) loss was 15.0 ± 8.7 (11.5% ± 6.1% of HB level loss). Multivariate linear regression analysis revealed that gender, hypertension, and albumin loss between preoperation and postoperation are influential factors of HBL in patients after LAG for gastric cancer. Compared to male patients, female patients are positively associated with HBL. CONCLUSION: In our study, we found HBL is a significant segment of TBL and is much larger than what we considered previously in LAG for gastric cancer. Gender, hypertension, and albumin loss are significantly correlated with HBL. Therefore, paying attention to HBL is significant for promoting clinical treatment and ensuring patients' safety.

SP1 reduces autophagic flux through activating p62 in gastric cancer cells.

Gastric cancer is the most common type of gastrointestinal cancer, causing mortality worldwide. However, the underlying molecular mechanism in gastric cancer progression remains unclear. The autophagic flux was determined in gastric cancer cells overexpressing or inhibiting Sp1 transcription factor (SP1) using western blotting, reverse transcription­polymerase chain reaction and immunofluorescence staining. Luciferase and ChIP assays were performed to detect the potential underlying mechanism of SP1 in gastric cancer cells. Lastly, immunohistochemistry was also performed on SP1 and p62 expression levels in human gastric cancer specimens. It was demonstrated that SP1 diminished autophagic flux via activating p62 in gastric cancer. Moreover, SP1 deficiency increased the rate of autophagy of gastric cancer cells. Notably, it was observed that SP1 enhanced the expression levels of p62 by directly binding to the promoter of p62. Analysis of gastric cancer specimen staining established that p62 expression levels were increased in SP1­positve gastric tissues. The present study provided evidence for a novel mechanism regulating autophagy in gastric cancer cells.

Compressed food with added functional oligopeptides improves performance during military endurance training.

BACKGROUND AND OBJECTIVES: Oligosaccharide or oligopeptide supplementation may have a significant impact on endurance performance. This study evaluated the effects of adding maltooligosaccharides (MO) or soy oligopeptides (SO) to compressed food (CF) on the physical response of soldiers to daily military training. METHODS AND STUDY DESIGN: Twelve soldiers were randomized to four diet groups: regular meals, CF, CFMO, and CFSO (crossover design). They participated in exercise tests including 90 minutes running at 55-65% VO2max and exhaustive running. Heart rates, rating of perceived exertion (RPE), and blood and urine samples were collected during exercise and recovery. RESULTS: The recovery heart rates were significantly lower with the CFMO diet compared with the other diets. Compared with all other diets, blood glucose levels were higher, post-exercise blood lactate levels were lower, and lactate clearance during recovery was higher with the CFMO diet, followed by the CFSO diet. Post-exercise levels of erythrocytes and hematocrit were significantly higher with the CFSO diet. Post-exercise urine specific gravity was lower with the CFMO diet and urine pH was decreased with the CFSO diet. Blood urea nitrogen (BUN) and uric acid (UA) were significantly higher with the CFSO diet than with the other diets. There was no significant difference in skeletal and cardiac muscle injury indices and RPE among diets. CONCLUSIONS: CFMO led to better heart rate recovery, improved and maintained blood glucose and increased removal of blood lactate. CFSO accelerated removal of blood lactate during recovery, maintained oxygen supply, and increased fluid retention.

Gossypol induces cell death by activating apoptosis and autophagy in HT-29 cells.

Gossypol is a polyphenolic, yellowish compound derived from cottonseed extract. The present study examined the effects of gossypol on the apoptosis and autophagy of HT­29 cells. A Cell Counting Kit­8 assay, Annexin V­FITC, JC­1 staining and western blotting were used to identify the viability of cells, stages of apoptosis and the expression levels of the signaling proteins. Gossypol promoted apoptosis and induced the loss of mitochondrial membrane potential. Further investigation of the apoptotic mechanism revealed that gossypol increased the ratio of B­cell lymphoma 2 (Bcl-2)-associated X protein/Bcl­2 protein levels and upregulated the expression of caspase­3. Gossypol also enhanced the activity of microtubule­associated protein light chain 3 LC3­II and Beclin­1 and downregulated LC3­I, in a dose­dependent manner. Together, these finding suggested that gossypol may be a novel and potential antitumor agent.

Romidepsin induces G2/M phase arrest via Erk/cdc25C/cdc2/cyclinB pathway and apoptosis induction through JNK/c-Jun/caspase3 pathway in hepatocellular carcinoma cells.

The aim of the study is to demonstrate the effect of Romidepsin in hepatocellular carcinoma (HCC) by inducing G2/M phase arrest via Erk/cdc25C/cdc2/cyclinB pathway and apoptosis through JNK/c-Jun/caspase3 pathway in vitro and in vivo. Human HCC cell lines were cultured with Romidepsin and DMSO (negative control) and 5-fluorouracil (positive control). Then the cells' viability and apoptosis were determined by cell proliferation assay and flow cytometry. Protein concentrations and expression changes were measured by Western blot. Subsequently, Huh7 cells were subcutaneously inoculated into the nude mice, which were employed to further probe the tumor-suppressive effect of Romidepsin in vivo. Romidepsin treatment led to a time- and dose-dependent induction of cell cycle arrest in the G2/M phase and apoptosis. G2/M phase arrest inhibited the proliferation of HCC cells by alterations in p21/cdc25C/cdc2/cyclinB proteins. Increased concentrations of Erk and JNK phosphorylations were observed in a dose-dependent manner in the Romidepsin group, but p38 phosphorylation was not affected. G2/M phase arrest and the apoptosis of HCC cells induced by Romidepsin were mediated by the activation of Erk/MAPK pathways and JNK/MAPK pathways. The tumor size was significantly larger in the negative control group compared to Romidepsin group and no significant loss in body weight was observed in the Romidepsin group. Our findings offer proof-of-concept for use of Romidepsin as a novel class of chemotherapy in the treatment of HCC.

Expression of Cytoplasmic 8-oxo-Gsn and MTH1 Correlates with Pathological Grading in Human Gastric Cancer.

BACKGROUND: Cancers have dysfunctional redox regulation resulting in production of reactive oxygen species (ROS), damaging DNA, RNA and free NTPs, and causing the accumulation of oxidative nucleic acids in cytoplasm. The major types are 8-oxo-7,8-dihydroguanine(8-oxoGsn) in RNA and 8-oxo-7,8-dihydro-2' deoxyguanosine(8-oxodGsn) in Mt-DNA. The MTH1 protein sanitizes oxidized nucleotide pools from NTPs to monophosphates, preventing the occurrence of transversion mutations. This study concerned cytoplasmic 8-oxodGsn/Gsn and MTH1 expression in gastric cancer and para-cancer tissues and elucidated roles of nucleic-acid oxidation and anti-oxidation. MATERIALS AND METHODS: A polymer HRP detection system was used to detect 8-oxo-Gsn/dGsn and MTH1 expression in 51 gastric cancer and para-cancer tissue samples. Analyses of patient clinical and pathological data were also performed. RESULTS: The expression of MTH1 and the 8-oxo-dGsn/Gsn ratio were significantly higher in cancer tissues than para-cancer tissues (P<0.05). Cytoplasmic 8-oxo-Gsn and MTH1 were both found to positively correlate (P<0.05) with tumor differentiation, while no significant associations were found with gender, age, invasion depth, lymph node metastasis and clinical stage (P>0.05). CONCLUSIONS: We found 8-oxo-dGsn/Gsn and MTH1 are both highly expressed in gastric cancer tissues, especially in well differentiated lesions. In addition, oxidated mtDNA is prevalently expressed in gastric cancers, while 8-oxo-Gsn expression in cytoplasmic RNA is a bit lower, but more selectively.

[Cloning and expression of aldolase encoding gene of Plasmodium falciparum FCC1/HN strain].

OBJECTIVE: To clone, sequence, and express the aldolase (ALD) encoding gene of Plasmodium falciparum FCC1/HN strain. METHODS: The ALD encoding gene was amplified by PCR from genomic DNA of FCC1/HN strain. The positive clones were screened and identified by agarose gel electrophoresis and endonuclease. The recombinant plasmid was transformed into E. coli M15. The fusion protein was expressed by IPTG induction and purified by Ni-NTA affinity chromatography and anion exchange column. RESULTS: The ALD gene of P. falciparum was amplified. Analysis of sequencing showed that the ALD gene of P. falciparum was identical with the sequence of other reported isolates. A Mr 41,000 fusion protein was induced by IPTG and was purified by chromatography. CONCLUSION: The ALD gene of P. falciparum FCC1/HN strain was identical to the other reported isolates. ALD fusion protein of P. falciparum was expressed and purified.