Autophagy Suppression Accelerates Apoptosis Induced by Norcantharidin in Cholangiocarcinoma.

Norcantharidin is a cantharidin demethylated analog with antitumor effects in many tumors, including cholangiocarcinoma. Autophagy suppression is known to increase chemosensitivity in cholangiocarcinoma. This study aimed to determine whether autophagy suppression accelerates apoptosis induced by norcantharidin in human cholangiocarcinoma cells. The human cholangiocarcinoma cell line QBC939 was incubated in RPMI 1640 medium with or without norcantharidin. Autophagy was induced using HBSS media with Ca2+ and Mg2+ supported by 10 mM HEPES or suppressed by treatment with 3-MA or transfection with siRNA against Atg5. The comparison was drawn between these conditions in mitochondrial membrane potential disturbance, the levels of reactive oxygen species (ROS), apoptotic proteins, and apoptosis. Cholangiocarcinoma cell apoptosis was accelerated by norcantharidin. Autophagy suppression up-regulated norcantharidin's pro-apoptotic effect, but autophagy induction weakened it. As apoptosis was accelerated, ROS production was up-regulated. Bax protein expression, cytochrome c levels and localization, mitochondrial membrane disturbance, and the levels of caspase-9, caspase-3, and cleaved PARP were higher when autophagy was suppressed, and all of those were down-regulated when autophagy was induced. To sum up, it was found that norcantharidin induced cholangiocarcinoma cell death, and autophagy suppression enhanced the pro-apoptotic action of norcantharidin, which appears to involve the mitochondrial apoptosis pathway activation and ROS generation.

Inhibition of autophagy enhances cinobufagin­induced apoptosis in gastric cancer.

Cinobufagin is a cardiotoxic bufanolide steroid secreted by the Asiatic toad Bufo gargarizans. Cinobufagin is one of the active ingredients in the anticancer Chinese medicine called Chan Su, which was demonstrated to be an effective treatment for gastric cancer. Increasing evidence shows that inhibition of autophagy has a pro­apoptotic effect on human gastric cancer cells. The aim of the present study was to investigate the relationship between cinobufagin, autophagy and apoptosis in gastric cancer. Autophagy was induced or inhibited in the human gastric cancer cell line SGC­7901 by incubation in HBSS media or by treatment with 3­methyladenine or ATG5 siRNA, respectively. Following treatment, the levels of apoptosis, apoptotic proteins, reactive oxygen species (ROS), and mitochondrial membrane potential were compared between the conditions. As anticipated, we found that cinobufagin increased apoptosis in SGC­7901 cells. Notably, inhibition of autophagy, monitored by the absence of the autophagosome marker LC3­II, also enhanced cell apoptosis. This effect was reversed when autophagy was induced by incubation in HBSS media. Enhanced expression of pro­apoptotic indicators, including BAX, cytosolic cytochrome c, cleaved PARP, caspase­3 and caspase­9, was detected when autophagy was suppressed. Increased pro­apoptotic protein expression was accompanied by disrupted mitochondrial membrane potential and elevated ROS production. Altogether, these data suggest that inhibition of autophagy enhances the anticancer action of cinobufagin through increased apoptosis of gastric cancer cells. Moreover, these effects may be partly mediated by ROS generation and the activation of the mitochondrial programmed cell death pathway.

MEK inhibitor enhances sensitivity to chemotherapeutic drugs in multidrug resistant hepatocellular carcinoma cells.

The aim of the present study was to investigate the association between the mitogen-activated protein kinase (MAPK) signal transduction pathway and multidrug resistance in hepatocellular carcinoma cells. A Cell Counting Kit-8 assay was used to determine the drug sensitivity of HepG2 and HepG2/ADM hepatocellular carcinoma cell lines in combination with the MAPK/extracellular-signal-regulated kinase kinase (MEK) inhibitor U0126. Flow cytometry was used to analyze the rate of apoptosis. The reverse transcription-quantitative polymerase chain reaction (RT-qPCR) was used to determine P-glycoprotein (P-gp) and multidrug resistance-associated protein 1 (MRP1) mRNA expression following treatment with various concentrations of U0126. P-gp and MRP1 expression levels were measured using Western blot analysis. The half-maximal inhibitory concentration was markedly decreased in combination with U0126. RT-qPCR results demonstrated that the expression of multidrug resistance 1 (MDR1) and MRP1 in HepG2/ADM cells was increased 5.37- and 6-14-fold compared with that in HepG2 cells. Furthermore, the expression levels in HepG2/ADM cells were decreased following U0126 treatment in a dose-dependent manner. The expression of P-gp and MRP1 in HepG2/ADM cells was increased 2.68- and 2.76-fold compared with that in HepG2 cells. Furthermore, the expression levels in HepG/ADM cells were decreased following U0126 treatment in a dose-dependent manner. The results of the present study indicate that the MEK inhibitor U0126 enhances sensitivity to chemotherapeutic drugs by downregulating P-gp and MRP1 expression in resistant hepatocellular carcinoma cells. The combination of MEK inhibitor and conventional chemotherapeutic drugs may provide novel therapeutic prospects for the treatment of drug-resistant hepatocellular carcinoma.

Recent perspectives of management of breast cancer metastasis - an update.

Breast cancer is the leading cause of cancer related deaths in women and one of the most common cancers globally. The major obstacle in the management of breast cancer, especially at advanced stages, is metastasis. Metastasis in the advanced stages of breast cancer could decrease survival to approximately 5 years. The reasons could include lack of targeted receptors or chemotherapeutic agents for the management of advanced-stage breast cancer metastasis. The new emerging avenues for the management of this deadly pathological state include local manipulations like radiofrequency ablation (RFA), microwave thermotherapy, cryosurgery (cryotherapy), chemoembolization, radioembolization, breast surgery, or metastasectomy. Few single-institution reports showed improved survival in selected patients like those with oligometastatic stage IV breast cancer. The present review article focused on these emerging new multimodality treatment approaches for a possible efficient management.

Clinical application of transanal ileal tube placement using X-ray monitoring.

Colorectal cancer, which includes colon and rectal cancer, is a common digestive tract tumor. Although surgery is the primary form of treatment, there are a number of drawbacks, including patients experiencing considerable pain and high cost. The present study was undertaken to examine the clinical value of transanal ileal tube placement under X-ray monitoring. Thirty-six cases of left colon obstruction presenting to our hospital between July 2011 and February 2014, underwent transanal ileal tube placement using a single-curve catheter guided by a guidewire under X-ray monitoring. An ileal tube was successfully inserted into 32 patients. Clinical symptoms were alleviated effectively within 48 h. Indwelling catheter decompression time was 4-9 days with an average of 5.61 days. In two cases, the colon guidewire perforated into the abdominal cavity. Repeated exploration resistance of the guidewire and catheter indicated stenosis at this position owing to obstruction. In conclusion, transanal placement of the ileal tube through X-ray monitoring is capable of effectively alleviating the symptoms of ileus. Thus, this constitutes a safe, effective, and economical method that is acceptable to patients.

Application of catheter and guidewire transmission technology in the reconstruction of gastroduodenal malignant stricture.

BACKGROUND: Gastroduodenal obstruction is a preterminal event in patients with advanced and recurrent malignancies of the stomach, pancreas, and duodenum. It severely limits the quality of life in affected patients due to constant vomiting and associated malnutrition. Gastroduodenal stent placement is a very safe and effective method in patients with unresectable malignant tumors. METHODS: Thirty-six patients with malignant gastroduodenal stenosis accompanied by gastrointestinal obstruction which was not traversable with a gastroscope were included in the study. Under X-ray monitoring, the catheter was inserted through the stenosis advancing with loach guidewire, which was later replaced by a stiff guidewire. A tri-lumen gastrojejunal catheter was then introduced using the stiff guidewire through the stenosis to perform gastrointestinal decompression and provide intestinal nutrition. In some patients, an intestinal stent was placed at the site of the stenosis. RESULTS: Intubation was successful in 35 patients out of 36. A total of 12 intestinal stents were successfully placed, including two at the pylorus, six at the site of gastrointestinal anastomosis, and four at the descending horizontal part of the duodenum. CONCLUSIONS: Establishing enteral nutrition via intubation using guidewire and catheter delivery technology is a simple, effective and safe strategy for patients with gastroscopic inaccessible malignant gastroduodenal stenosis, and it is recommended in hospital practice.

Laparoscopic jejunoileal side-to-side anastomosis for the treatment of type 2 diabetes mellitus in Chinese patients with a body mass index of 24-32 kg/m2.

OBJECTIVE: Laparoscopic jejunoileal side-to-side anastomosis (LJISSA) is an upcoming procedure that offers good metabolic improvement without causing significant malabsorption. The objective of this study was to evaluate the results of this novel procedure for the control of type 2 diabetes mellitus (T2DM) in patients with a body mass index (BMI) of 24-32 kg/m 2. MATERIALS AND METHODS: Fifty-seven patients with T2DM who underwent LJISSA between February 2010 and May 2013 were recruited in this study. Data collected included fasting blood glucose (FBG), 2 h postprandial blood glucose (2 h PBG), 1 h postprandial C peptide (1 h C-P), and glycosylated hemoglobin (HbA1c). RESULTS: Postoperatively, glycemic parameters (FBG and 2 h PBG, HbA1c and 1 h C-P) improved in all 57 patients. At 12 months, 34 patients had a remission of diabetes, and the remaining 23 patients showed a significantly decreased requirement for oral hypoglycemic agents. The patients with a BMI of 28-32 kg/m 2 had significant weight loss of between 7.8% and 20% (P < 0.05), whereas weight loss was not significant in those with a BMI of 24-28 kg/m 2. The group achieving remission had a higher BMI (28-32 kg/m 2), shorter duration of diabetes (<10 years), and higher stimulated C-P (>4 ng/mL). These three factors may be the predictors of diabetes resolution at 12 months. CONCLUSION: LJISSA seems to be a promising procedure for the control of T2DM. A multicenter study with a larger number of patients and a longer follow-up period is needed to substantiate our preliminary findings.

Efficacy of intestinal decompression with long nasointestinal tube and selective contrast radiography in the treatment of small bowel obstruction in elderly patients.

BACKGROUND: This study's aim was to assess the efficacy of intestinal decompression with long tube and selective intestinal radiography in the diagnosis and treatment of small bowel obstruction (SBO) in elderly patients. METHODS: Thirty-two elderly patients with SBO received intestinal decompression with a 300-cm long nasointestinal tube inserted into upper jejunum under radiographic control. The long tube was passed into the proximal part of obstruction or the proximal end of ileum driven by intestinal peristalsis. Selective contrast radiography was done using direct injection of double-contrast medium consisting of 20-100 mL of 76% gastrografin and 50-200 mL of air. The dynamic and multi-position radiographic observation was conducted. RESULTS: Intubation was successful in all 32 patients. SBO resolution was successful in 29/32 (90.6%) patients. The 3 remaining patients proceeded to undergo surgery. Radiographic findings showed no obvious abnormalities in 25/32 (78.1%) patients, adhesive SBO in 6/32 (18.6%), and metastatic intestinal tumor in 1/32 (3.1%) patient. CONCLUSIONS: Decompression using long tube can quickly and effectively relieve obstructive symptoms in elderly patients, and help to avoid emergency surgery and to resolve obstruction. Concurrent contrast radiography is helpful to verify the location and degree of obstruction, and to reveal the cause of the obstruction.

Atg5 siRNA inhibits autophagy and enhances norcantharidin-induced apoptosis in hepatocellular carcinoma.

Cantharidin is a terpenoid isolated from Chinese blister beetles, and norcantharidin (NCTD) is a demethylated analog of cantharidin. It has been reported that cantharidin and norcantharidin have anticancer activities. Growing evidence suggests that inhibiting autophagy can induce apoptosis in the human hepatoma cell line HepG2. The objective of the present study was to determine whether inhibition of autophagy enhances NCTD-induced apoptosis in HepG2 cells. HepG2 cells were cultured in DMEM containing NCTD. Autophagy was upregulated in the presence of HBSS media supplemented with Ca2+ and Mg2+ and 10 mM HEPES and downregulated in the presence of 3-methyladenine (3-MA) and Atg5 siRNA. Autophagy, cell viability, and the expression of apoptotic proteins were assessed in HepG2 cells. Our data showed that cell apoptosis generally increased after norcantharidin treatment in HepG2 cells. Expression of LC3-II, an autophagosome marker, increased when cells were treated with HBSS media. It also increased cell viability. However, in the presence of 3-MA and Atg5 siRNA, autophagy was inhibited, LC3-II expression decreased and cell apoptosis increased. There was increased expression of Bax, cytochrome c, cleaved caspase-3, caspase-9 and PARP and the mitochondrial membrane potential was disrupted. Additionally, increased apoptosis was accompanied by increased reactive oxygen species (ROS) production. NCTD has anticancer activity, and Atg5 siRNA-mediated downregulation of autophagy enhanced its anticancer actions due to ROS generation and activation of the mitochondrial apoptosis pathway.

[Feasibility of terminal intestinal exteriorization in laparoscopic anterior resection for anterior cancer].

OBJECTIVE: To discuss the feasibility of terminal intestinal exteriorization (exteriorization without ileostomy) in laparoscopic anterior resection for rectal cancer. METHODS: Clinicopathological data of 77 patients undergoing laparoscopic anterior resection for low rectal cancer in our department from January 2011 to December 2013 were retrospectively analyzed. After laparoscopic rectal resection, 32 patients received terminal intestinal exteriorization (exteriorization group) and 45 patients received preventive ileostomy (ileostomy group). Anastomosis-related, stoma-related and intestinal stoma closure-related morbidity was compared between the two groups. RESULTS: There were no significant differences in operative time, blood loss and overall hospital stay between the two groups (all P>0.05). The total hospital cost was (5.39±1.74)×10(4) yuan in the exteriorization group, and (6.98±1.37)×10(4) yuan in the ileostomy group(P<0.01). The incidences of postoperative anastomotic fistula was not significantly different between the two groups(P>0.05). Three patients(9.4%) developed anastomotic leak in the exteriorization group and 2(4.4%) in the ileostomy group. The anastomotic leak was managed by opening the external intestinal wall and maturating an ileostomy under local anaesthesia. All these 5 patients were cured with nutritional support, antibiotics, continuous local drainage. In the exteriorization group, 5 patients had complications related to stoma and intestinal stoma closure operation(15.6%), which was lower than(42.2%) in the ileostomy group(P=0.013). CONCLUSION: Terminal intestinal exteriorization in laparoscopic anterior resection is a safe and feasible surgical procedure with little trauma and less hospital cost, which can be an alternative as a prophylactic treatment for patients with high risk of anastomotic leak.

Radical Gastrectomy Combined with Modified Gastric Bypass Surgery for Gastric Cancer Patients with Type 2 Diabetes.

The aim of the study is to explore the effectiveness of radical gastrectomy with modified gastric bypass surgery in treating gastric cancer patients with type 2 diabetes mellitus (T2DM). A total of 93 patients with gastric cancer and T2DM were treated in our hospital and enrolled in this study. Patients in group A (n = 30) had a body mass index (BMI) of >28 kg/m(2). Radical total gastrectomy and modified esophagojejunal Roux-en-y anastomosis were performed on 13 patients, and radical distal subtotal gastrectomy and gastric remnant jejunal Roux-en-y anastomosis were performed on 17 patients. The data from groups B, C, and D were derived from 63 patients with gastric cancer and diabetes who were admitted to our hospital from January 2005 to July 2012. All patients underwent radical gastrectomy (including 21 cases of gastric cancer surgery with Billroth I anastomosis, 25 cases of radical gastrectomy with Roux-en-Y anastomosis and BMI >28 kg/m(2), and 17 cases with BMI <28 kg/m(2)). The BMI, fasting blood glucose (FBG), meal after the 2-hour glucose (2 h PBG), C-peptide (C-P), and glycosylated hemoglobin (HbAIC) data were collected before and 6 and 12 months after surgery. In groups A and D, BMI, FBG, 2 h PBG, C-P, and HbAIC at the 6th and 12th post-operative months were significantly lower than those before the surgery. In group B, BMI, FBG, 2 h PBG, C-P, and HbAIC at the 6th and 12th post-operative months did not decrease significantly, when compared with the pre-operative levels. In group C, BMI, FBG, 2 h PBG, C-P, and HbAIC at the 6th and 12th post-operative months decreased but showed no statistical significance. However, in comparison, groups A C showed significant differences after the surgeries. Radical gastrectomy combined with modified gastric bypass surgery is effective in treating patients with gastric cancer with type 2 diabetes, although this requires further investigation.

ZnPP reduces autophagy and induces apoptosis, thus aggravating liver ischemia/reperfusion injury in vitro.

There is growing evidence indicating that autophagy plays a protective role in liver ischemia/reperfusion (IR) injury. Heme oxygenase-1 (HO-1) can also prevent liver IR injury by limiting inflammation and inducing an anti-apoptotic response. Autophagy also plays a crucial role in liver IR injury. The aim of the present study was to investigate the role of HO-1 in liver IR injury and the association between HO-1, autophagy and apoptotic pathways. IR simulation was performed using buffalo rat liver (BRL) cells, and HO-1 activity was either induced by hemin (HIR group) or inhibited by zinc protoporphyrin (ZnPP) (ZIR group). In the HIR and ZIR group, the expression of HO-1 and autophagy-related genes [light chain 3-Ⅱ (LC3-Ⅱ)] was assessed by RT-qPCR and the protein expression of caspases, autophagy-related genes and genes associated with apoptotic pathways (Bax) was detected by western blot anlaysis. The results of RT-PCR revealed the genetically decreased expression of HO-1 and autophagy-related genes in the ZIR group. Similar results were obtained by western blot analysis and immunofluorescence. An ultrastructural analysis revealed a lower number of autophagosomes in the ZIR group; in the HIR group, the number of autophagosomes was increased. The expression of Bax and cytosolic cytochrome c was increased, while that of Bcl-2 was decreased following treatment of the cells with ZnPP prior to IR simulation; the oppostie occurred in the HIR group. Cleaved caspase-3, caspase-9 and poly(ADP-ribose) polymerase (PARP) protein were activated in the IR and ZIR groups. The disruption of mitochondrial membrane potential was also observed in the ZIR group. In general, the downregulation of HO-1 reduced autophagy and activated the mitochondrial apoptotic pathway.

The clinical value of Fufangkushen injection in the treatment of stomach cancer: a meta-analysis.

OBJECTIVE: The purpose of this meta-analysis was to evaluate the effects of Fufangkushen injection combined with chemotherapy in the treatment of stomach cancer. MATERIALS AND METHODS: The relevant clinical trials about Fufangkushen injection combined with chemotherapy in the treatment of stomach cancer were search in the data bases of Pubmed, EMBASE, Cochran and CNKI. The data related to objective response rate, Karnofsky (KPS) score and toxicity were extracted and pooled using the Stata 11.0 software. Dichotomous data was presented as risk ratio (RR) and its 95% confidence interval (95% CI). RESULTS: Thirteen relevant trials were included in this meta-analysis. Heterogeneity test indicated there was no statistical heterogeneity among the studies, thus the fixed effects mode was used to calculat the results. Pooled results indicated that the objective response rate (ORR) and KPS score improvement in Fufangkushen chemotherapy group was significant higher than that of control group (RR = 1.24, P < 0.05). Synthesis data also demonstrated the Fufangkushen injection can significantly decrease the risk of developing granulocytopenia in stomach cancer patients treated with chemotherapy (RR = 0.67,P < 0.05). CONCLUSION: Fufangkushen injection combined with chemotherapy can increase the objective response rate, improve the quality of life and decrease the risk of developing granulocytopenia in patients with stomach cancer.

In vitro and in vivo evaluation of the antitumor efficiency of resveratrol against lung cancer.

Lung cancer remains a deadly disease with unsatisfactory overall survival. Resveratrol (Res) has the potential to inhibit growth of several types of cancer such as prostate and colorectal examples. In the current study, we evaluated in vitro and in vivo anticancer efficiency of Res in a xenograft model with A549 cells. Cell inhibition effects of Res were measured by MTT assay. Apoptotis of A549 cells was assessed with reference to caspase-3 activity and growth curves of tumor volume and bodyweight of the mice were measured every two days. In vitro cytotoxicity evaluation indicated Res to exert dose-dependent cell inhibition effects against A549 cells with activation of caspase-3. In vivo evaluation showed Res to effectively inhibit the growth of lung cancer in a dose- dependent manner in nude mice. Therefore, we believe that Res might be a promising phytomedicine for cancer therapy and further efforts are needed to explore this potential therapeutic strategy.

Effects of 5-aminolevulinic acid-mediated photodynamic therapy on antibiotic-resistant staphylococcal biofilm: an in vitro study.

BACKGROUND: Treatments of infections are not always successful because of multi-antibiotic-resistant organisms. It is therefore particularly urgent to provide more effective anti-infective strategy against these organisms. 5-Aminolevulinic acid (ALA), with the chemical structure C5H9NO3, is the only photodynamic therapy agent that is a biochemical precursor of a photosensitizer (protoporphyrin IX [PpIX]), which is naturally produced by the body. 5-Aminolevulinic acid-mediated photodynamic therapy (ALA-PDT) has been shown to have a strong effect on the treatment of localized cancerous and precancerous lesions, and further study demonstrated its efficacy for gram-positive and gram-negative bacteria. However, its effect on biofilm formed by antibiotic-resistant strains has not been reported. METHODS: In this study, we evaluated the effectiveness of ALA-PDT on biofilms formed by methicillin-resistant Staphylococcus aureus (ATCC 43300) and methicillin-resistant S epidermidis (MRSE 287). The strains were cultured with 40 mM of ALA in 24-well microtiter plates containing coverslips at 37°C for 24 h in the dark. PpIX fluorescence in biofilms formed by the two strains was observed by confocal laser scanning microscopy (CLSM). ALA-treated biofilms were irradiated at different doses (0, 100, 200, and 300 J/cm(2)) using a semiconductor laser. Biofilm exposed only to Tryptone Soy Broth or irradiation (300 J/cm(2)) was investigated. Viability determination, CLSM, and scanning electron microscopy were used to investigate the photodynamic inactivation of ALA-PDT. RESULTS: ALA was absorbed and converted to PpIX by both methicillin-resistant S aureus and methicillin-resistant S epidermidis. No cell inactivation was detectable in biofilms of either strain incubated with ALA without exposure to light, incubated with Tryptone Soy Broth only, or irradiated with red light only. However, a significant number of cells within biofilms were inactivated during irradiation with different doses of red light in the presence of 40 mM of ALA in a dose-dependent manner. The drastic reduction in cell survival within biofilms and the disruption of biofilms were confirmed by CLSM and scanning electron microscopy. CONCLUSIONS: ALA-PDT has the potential to eliminate the biofilm of Staphylococcus, especially antibiotic-resistant strains, effectively. It will be suitable for the treatment of superficial local infections such as surface wounds, burns, oral and dental infections, dermatologic infections such as acne and rosacea, and soft tissue and bone infections with bone exposure.