[Effect of electroacupuncture on colonic autophagy and AMPK/mTOR signaling pathway in rats with acute ulcerative colitis].

OBJECTIVE: To observe the effect of electroacupuncture （EA） at "Zhongwan" （CV12）, "Tianshu" （ST25） and "Shangjuxu" （ST37） （an acupoint prescription "Changbingfang" for treatment of intestinal disorders） on autophagy and expression of AMPK/mTOR signaling pathway in rats with ulcerative colitis （UC）, so as to explore its mechanism underlying improvement of UC. METHODS: Thirty-two male SD rats were randomly divided into control, model, medication and EA groups, with 8 rats in each group. The UC model was established by free drinking of 5% dextran sulfate sodium salt solution for 7 days. EA stimulation （10 Hz/50 Hz） was delivered to CV12, ST25 and ST37 for 20 min, once a day for 3 consecutive days. Rats of the medication group received gavage of mesalazine suspension （200 mg/kg） once a day, 3 times in total. The rats' general conditions were recorded for calculating the disease activity index （DAI） score （0-4 points）. Histomorphological changes of colon were observed via HE staining. The levels of serum interleukin 6 （IL-6）, tumor necrosis factor-α （TNF-α） and IL-10 were measured by ELISA. The mRNA expressions of LC3B and p62 were tested by fluorescence quantitative PCR. Western blot was used to detect the expression levels of LC3B, p62 and AMPK/mTOR pathway related proteins in colon tissues. RESULTS: Compared with the control group, the DAI score, contents of serum IL-6 and TNF-α, the expression levels of p62 protein and mRNA, ratio of p-mTOR/mTOR were significantly increased （P<0.01）； while the content of serum IL-10, the expression levels of LC3B mRNA, ratio of LC3BⅡ/LC3BⅠ and p-AMPK/AMPK were decreased （P<0.01, P<0.05） in the model group. Relevant to the model group, modeling-induced increases of DAI score, serum IL-6, TNF-α and IL-10 contents, expressions of p62 protein and mRNA, LC3B mRNA, ratio of p-mTOR/mTOR, LC3BⅡ/LC3BⅠ and p-AMPK/AMPK were reversed in both medication and EA groups （P<0.01, P<0.05）. The effect of EA was apparently superior to that of mesalazine in up-regulating ratio of LC3BⅡ/LC3BⅠ and p-AMPK/AMPK, p62 mRNA expression （P<0.01, P<0.05）, and in down-regulating ratio of p-mTOR/mTOR （P<0.05）. H.E. staining showed severe damage of the colonic mucosal barrier with infiltration of a large number of inflammatory cells in the model group, which was milder in medication and EA groups. CONCLUSION: EA of acupoint recipe "Changbingfang" can improve the symptoms in UC rats, which may be related to its functions in promoting colonic autophagy, increasing AMPK phosphorylation level, and decreasing mTOR phosphorylation level.

Retraction: MicroRNA-371-5p targets SOX2 in gastric cancer.

[This retracts the article DOI: 10.18632/oncotarget.8289.].

MicroRNA-371-5p targets SOX2 in gastric cancer.

We evaluated miR-371-5p expression in gastric cancer (GC) tissues and its influence on the expression of downstream genes, especially SOX2. MiR-371-5p expression (measured using qRT-PCR) was upregulated in GC tissues and correlated positively with TNM staging and lymph node (LN) metastasis. MiR-371-5p expression was higher in human GC cell lines (AGS, MKN-28, BGC-823, MGC-803, SGC-7901 and MKN-45) than in human normal gastric epithelial (GES-1) cells (all P < 0.05). MGC-803 tumor cell growth (measured with an MTT assay), migration, and invasion (measured with Transwell chamber assays) were severely inhibited in cells transfected with a miR-371-5p inhibitor, whereas they were stimulated in cells transfected with SOX2 siRNA or miR-371-5p inhibitor + SOX2 siRNA. Expression of SOX2 mRNA and protein (assessed with qRT-PCR and Western blot) were greatly enhanced in the miR-371-5p inhibitor group. These results indicate that miR-371-5p expression is strongly upregulated in GC tissues and negatively correlated with SOX2 expression, while miR-371-5p expression is inversely related to proliferation, TNM stage, and LN metastasis of GC cells. Suppression of miR-371-5p may inhibit the growth and invasion of MGC-803 GC cells by upregulating SOX2 expression.

Folate-decorated anticancer drug and magnetic nanoparticles encapsulated polymeric carrier for liver cancer therapeutics.

Nanoparticulate system with theranostic applications has attracted significant attention in cancer therapeutics. In the present study, we have developed a novel composite PLGA NP co-encapsulated with anticancer drug (sorafenib) and magnetic NP (SPION). We have successfully developed nanosized folate-conjugated PEGylated PLGA nanoparticles (SRF/FA-PEG-PLGA NP) with both anticancer and magnetic resonance property. We have showed that FA-conjugated NP exhibits sustained drug release and enhanced cellular uptake in BEL7402 cancer cells. The targeted NP effectively suppressed the tumor cell proliferation and has improved the anticancer efficacy than that of free drug or non-targeted one. Additionally, enhanced MRI properties demonstrate this formulation has good imaging agent characteristics. Finally, SRF/FA-PEG-PLGA NP effectively inhibited the colony forming ability indicating its superior anticancer effect. Together, these multifunctional nanoparticles would be most ideal to improve the therapeutic response in cancer and holds great potential to be a part of future nanomedicine. Our unique approach could be extended for multiple biomedical applications.

Aberrant promoter methylation of the vimentin gene may contribute to colorectal carcinogenesis: a meta-analysis.

This meta-analysis of published cohort studies was conducted to evaluate how closely the promoter methylation of the vimentin gene is correlated with the pathogenesis of colorectal carcinogenesis (CRC). The Web of Science (1945 ~ 2013), Cochrane Library Database (issue 12, 2013), PubMed (1966 ~ 2013), EMBASE (1980 ~ 2013), CINAHL (1982 ~ 2013), and Chinese Biomedical Database (CBM) (1982 ~ 2013) were searched without language restrictions. Meta-analyses were conducted using Stata software (Version 12.0, Stata Corporation, College Station, TX, USA). Odds ratios (ORs) and 95 % confidence intervals (95 %CI) were calculated. Seven clinical cohort studies with a total of 467 CRC subjects met our inclusion criteria. Our meta-analysis results demonstrated that the frequency of vimentin promoter methylation in cancer tissues was significantly higher than in normal and benign tissues (cancer tissues vs. normal tissues: OR = 32.41, 95 %CI = 21.04 ~ 49.93, P < 0.001; cancer tissues vs. benign tissues: OR = 1.60, 95 %CI 1.05 ~ 2.42, P = 0.028). Ethnicity-stratified analysis indicated that the frequency of aberrant vimentin promoter methylation was correlated with the pathogenesis of CRC in both Asians and Caucasians. The findings of our meta-analysis confirm that vimentin methylation may play a crucial role in the pathogenesis of CRC.

[Clinicopathological significance of the expression of carbonic anhydrase II in human pancreatic invasive ductal cancer].

OBJECTIVE: To study the clinicopathological significance of the expression of carbonic anhydrase (CA)II protein and mRNA in primary invasive ductal cancer (IDC) of human pancreas. METHODS: The expression of CAII protein in 33 paired paraffin embedded IDC specimens of the pancreas and paired adjacent non-cancerous pancreatic tissues was detected by immunohistochemistry. Western blot and reverse transcription polymerase chain reaction (RT-PCR) were used to examine the expression of CAII protein and mRNA level in 12 paired fresh IDC specimens of the pancreas and adjuvant non-cancerous pancreatic tissues. The relationship between the protein expression and clinicopathological features was analyzed. RESULTS: Overexpression of CAII protein was shown in 11 cases of pancreatic IDC tissues (33.3%, 11/33), which was much lower than that in paired non-cancerous pancreatic tissues (72.7%, t = 6.275, P = 0.000). The expression of CAII protein had no correlation with tumor position (χ² = 0.992, P = 0.319), differentiation (χ² = 0.866, P = 0.352), TNM stage (χ² = 1.210, P = 0.271) and Lymph node metastasis (χ² = 0.798, P = 0.372), but had bordering statistic sig with the prognosis of the patients (χ² = 3.233, P = 0.072). The median survival time in the patients with high expression of CAII protein was 540 days, while that in the patients with low expression was 320 days. The expression of CAII protein and mRNA was lower in IDC than that in paired non-cancerous pancreatic tissues detected by Western blot and RT-PCR respectively (t = 3.399, P = 0.006; t = 2.281, P = 0.043). CONCLUSION: CAII is down regulated in pancreatic IDC and might be relative with the prognosis.

[Clinicopathological significance of the expression of carbonic anhydrase I and II in human pancreatic cancer].

OBJECTIVE: To explore the clinicopathological significance of the expression of carbonic anhydrase I (CAI) and carbonic anhydrase II (CAII) protein and mRNA levels in pancreatic ductal adenocarcinoma (PDAC). METHODS: The expression of CAI and CAII protein in 57 pairs of paraffin-embedded PDAC specimens and adjacent non-cancerous pancreatic tissues were detected by immunohistochemistry. The relationship between the protein expression and clinicopathological characters was analyzed. Western blot and quantitative real-time polymerase chain reaction (QRT-PCR) were used to examine the expression of CAI and CAII protein and mRNA levels in 16 paired fresh PDAC specimens, adjacent non-cancerous pancreatic tissues and 3 different differentiated pancreatic cancer cells (PANC-1, BxPC-3 & SW1990). RESULTS: The expression of CAI protein was higher in PDAC compared with that in paired non-cancerous pancreatic tissues (t = 2.395, P = 0.020), whereas, CAII expression was significantly lower in PDAC than that in paired non-cancerous pancreatic tissues (t = 4.296, P = 0.000). The expression of CAI and CAII protein had a positive correlation with tumor differentiation (χ(2) = 7.557, P = 0.023; χ(2) = 7.822, P = 0.020) and CAI showed a direct negative correlation with vascular invasion (χ(2) = 6.349, P = 0.012). Univariate and multivariate analysis with clinicopathological characters revealed that the CAII expression was an independent prognostic indicator for PDAC patients (P = 0.017; P = 0.011 respectively). No significant difference of CAI mRNA and protein expression existed between PDAC and adjacent normal pancreatic tissues (t = 1.619, P = 0.126; t = 1.352, P = 0.197) as detected by Western blot and QRT-PCR, but the expression of CAII mRNA and protein levels was much lower in PDAC than that in paired non-cancerous pancreatic tissues, respectively (t = 3.360, P = 0.004; t = 2.934, P = 0.010). Moreover, the mRNA and protein expression of CAI and CAII gradually increased with the better differentiation degree of pancreatic cancer cells. CONCLUSIONS: CAI is up-regulated and CAII down-regulated in PDAC. Both of them are correlated with tumor differentiation. CAII expression is an independent prognostic indicator for PDAC patients.

[Analysis of pancreatic leaking-related risk factors after pancreaticoduodenectomy].

OBJECTIVE: To analysis the risk factors of pancreatic fistula after pancreaticoduodenectomy (PD). METHODS: A retrospective clinical study had been done in 97 patients who underwent PD between June 2001 and June 2006. The two groups were first compared by the univariate analysis;logistic regression was then used to determine the effect of multiple factors on pancreatic fistula. A P-value of less than 0.05 was considered to be statistically significant. RESULTS: Of the 97 patients, 13 patients were identified as having pancreatic fistula. Factors significantly increasing the risk of pancreatic fistula by univariate analysis included preoperative serum total bilirubin (P = 0.038), operative time (P = 0.003) and whether or not Braun anastomosis (P = 0.034), and prophylactic use of somatostatin (P = 0.003) after operation. A multivariate logistic regression analysis revealed the factors most highly associated with pancreatic fistula to be preoperative serum total bilirubin (OR = 11.687, P = 0.021) and postoperative prophylactic use of somatostatin (OR = 0.056, P = 0.020). CONCLUSIONS: Preoperative serum total bilirubin more than 170 mmol/L was a risk factor of pancreatic fistula after PD, and postoperative prophylactic use of somatostatin was a protect factor of pancreatic fistula after PD.

[Surgical treatment of asymptomatic primary hyperparathyroidism].

OBJECTIVE: To discuss the diagnosis and management of asymptomatic primary hyperparathyroidism (PHPT). METHODS: Clinical data of 46 cases of primary hyperparathyroidism from January 1990 to December 2006 were retrospectively analyzed. There were 5 cases of asymptomatic PHPT. Three out of the 5 cases obtained the diagnosis by routine health examination and 1 case was misdiagnosed as thyroid tumor before surgery, but was conformed as parathyroid adenoma by intraoperative biopsy. Remaining 1 case was diagnosed because of weakness. The serum calcium and the parathyroid hormone (PTH) levels were elevated in 4 cases, while only 1 being normal range. Unilateral neck exploration was performed in all 5 cases. RESULTS: There were no operational death, recurrent nerve injury or other complications. All patients had the same pathological diagnosis as parathyroid adenomas. Three cases showed gentle circumoral paresthesia after surgery with normal serum level of calcium, but the symptoms were relieved with oral use of calcium gluconate. Only 1 patient had tetany with the lowest level of serum calcemia at 1.96 mmol/L in 24 h postoperatively. The signs and symptoms were all relieved by intravenous use of calcium gluconate for 3 d after surgery. Remaining 1 case has normal level of serum calcemia after surgery. Time range of following-up for 4 cases was from 2 months to 2 years. The level of serum calcemia was normal for them. One lost following-up. CONCLUSIONS: Asymptomatic primary hyperparathyroidism could be diagnosed according to co-elevated serum calcemia and PTH without typical symptoms. Unilateral neck exploration was the best choice for the patients with accurate imaging localization. Conservative management including adequate hydration, dietary calcium intake and pharmacological approaches could be used for the patients who were unfit for surgery.

Promoter hypermethylation of death-associated protein kinase gene in cholangiocarcinoma.

BACKGROUND: Death-associated protein kinase (DAPK) is a Ca2+/calmodulin-regulated Ser/Thr kinase which is involved in apoptosis. The aberrant methylation of its promoter region CpG islands may be one of the important mechanisms of carcinogenesis. We studied the relationship of methylation status and expression of the DAPK gene with the clinical findings in cholangiocarcinoma. METHODS: Target DNA was modified by sodium bisulfite, coverting all unmethylated, but not methylated, cytosines to uracil, and subsequently detected by methylation-specific PCR. Moreover, mRNA expression of the DAPK gene was assessed by RT-PCR. RESULTS: Aberrant methylation of the DAPK gene was detected in 11 (30.6%) of 36 tissue specimens of cholangiocarcinoma, and in 2 (5.6%) of 36 specimens of adjacent normal tissues. DAPK mRNA was not expressed in tumor and adjacent tissues with hypermethylation of the DAPK promoter. There were no statistical differences in the extent of differentiation and invasion, lymph node metastasis or pathologic type between the methylated and unmethylated tissues. CONCLUSIONS: The frequency of DAPK gene methylation in cholangiocarcinoma is high and it may offer an effective means for earlier auxiliary diagnosis of the malignancy. The DAPK gene is probably suppressed by methylation, and it could become resistant to apoptosis and immunological surveillance. The DAPK gene epigenetically affected by methylation may be associated with the carcinogenesis of cholangiocarcinoma.

Giant mucinous biliary cystadenoma: a case report.

BACKGROUND: Biliary cystadenoma is a very rare cystic neoplasm of the liver. Its clinical features, diagnosis, pathologic characteristics, and optimal surgical management have not been defined clearly. In this article we describe the details of this rare disease. METHODS: A 40-year-old woman with a mass of the liver was verified by ultrasonography and LT. Ultrasonography showed a mixed echo of 18.4 cm x 14.72 cm x 15.54 cm in the left lobe of the liver. CT showed a vesicula of 19.9 cm x 13.5 cm in the right epigastrium, with a low density, clear edge, uneven density, and calcified shadow. The patient received successfully a left hepatectomy. Laboratory examination showed an elevation of CA125 to 62.62 U/ml and CA199>1000 U/ml. RESULTS: After the left hepatectomy, the patient was fully recovered. Her biliary cystadenoma was characterized by specific histological findings. During operation, a large cystic lesion was seen in the left hepatic lobe; its surface was dark red with abundant blood supply. Gross examination showed that the tumor almost occupied. The whole left lobe with a small amount of normal liver tissue close to the deltoid ligament. Pathologically, additional lobulated spaces were seen in the tumor with a lot of mucusa. The interior wall was lined with bile duct tissue, indicating a benign mucinous biliary cystadenoma. CONCLUSIONS: Ultrasonography and CT are the major methods for the diagnosis of mucinous biliary cystadenoma liver. Operation is the best way of treatment.

[Epidermal growth factor-mediated NF-kappaB activation promotes uPA expression and invasiveness in pancreatic cancer cells].

OBJECTIVE: To determine the effect of EGF on the invasiveness of pancreatic cancer cells and its related regulatory mechanism. METHODS: The effects of EGF on the proliferation, adhesion and invasion of pancreatic cancer cells were detected by WST-1 proliferation assay, adhesion assay and invasive assay. The expression of uPA was assayed by Western blot and RT-PCR. The activity of NF-kappaB was examined by EMSA. RESULTS: EGF significantly increased the invasiveness of pancreatic cancer cells but did not affect cell proliferation or adhesion. Increased invasiveness was associated with the induction of uPA at both mRNA and protein levels. Furthermore, EGF stimulated the NF-kappaB binding activity, and pretreatment of cells with a NF-kappaB inhibitor, pyrrolidine dithiocarbamate, markedly attenuated EGF-induced NF-kappaB activation. Subsequently, the EGF-induced uPA expression and invasiveness were also inhibited by NF-kappaB inhibitor. CONCLUSION: Our findings indicated that NF-kappaB-mediated up-regulation of uPA expression is responsible for EGF-induced invasiveness in pancreatic cancer cells, and implicate that such anti-NF-kappaB therapy with NF-kappaB inhibitors may contribute to the reduction of invasiveness of pancreatic cancer.

[Comparison of pBcl-2 and pBax expression in primary invasive ductal pancreatic cancer between Chinese and Japanese patients].

OBJECTIVE: To clarify the clinicopathologic significance of the expression of the Bcl-2 protein (pBcl-2) and the Bax protein (pBax), and their clinical implications in Chinese and Japanese patients with human invasive ductal carcinomas (IDCs) of the pancreas. METHODS: The study included 59 Chinese and 65 Japanese patients with IDCs of the pancreas. pBcl-2 and pBax expression were immuno-stained with streptavidin-biotin (SAB) method. RESULTS: pBcl-2 (+) was seen in 35.6% of Chinese and in 23.1% of Japanese patients. pBax (+) was seen in 49.2% of Chinese and 64.7% of Japanese patients. A comparison between them showed that there were significant differences in the male patients, in the patients with the moderately differentiated cancer, and in the elderly patients (chi squared = 4.447, P = 0.035; chi squared = 4.114, P = 0.043; chi squared = 6.657, P = 0.010 respective). In both Chinese and Japanese patients, those with pBcl-2 positive expression had a significantly higher survival rate than those with negative one (chi squared = 9.99, P = 0.0016; chi squared = 7.63, P = 0.0058). The group with pBax positive expression had a significantly higher survival rate in Japanese patients (chi squared = 9.37, P = 0.0022). Japanese patients whose tumors exhibited pBcl-2 and pBax positive immunostaining survived significantly longer than Chinese patients did (chi squared = 4.48, P = 0.0342; chi squared = 5.23, P = 0.023). CONCLUSIONS: The expressions of both pBcl-2 and pBax are high found in Chinese and Japanese patients. The pBcl-2 positive expression implies a better prognosis in both Chinese and Japanese patients with IDCs of the pancreas. The effect of pBax expression on prognosis is different between Chinese and Japanese patients.

[Expressions of p53 and Gadd45a proteins in human pancreatic cancer and their clinicopathological significance].

OBJECTIVE: To study the expressions of p53 and Gadd45a proteins and their clinicopathological significance in human pancreatic cancer. METHODS: The expression of p53 and Gadd45a proteins was detected with immunohistochemistry in a series of 59 pancreatic cancers. Their relationships with the clinicopathological parameters including gender, tumor site, TNM stage, histological differentiation, and the prognosis of pancreatic cancer patients were analyzed. RESULTS: The positive expression rate of p53 protein was 67.8% (40/59) and that of Gadd45a protein was 42.4% (25/59). The positive expression rate of p53 protein is significantly higher in patients < 65 years than in patients > or = 65 years (chi squared = 4.711, P = 0.030). Gadd45a expression was not correlated to the age of the patients. No significant difference was found between the expression of p53 proteins and histological differentiation and TNM stage of the tumors. Gadd45a expression was correlated with histological differentiation of pancreatic cancer (chi squared = 10.052, P = 0.007), but not with TNM stage of the tumors. No significant differences in the prognosis were found between the groups with and without p53 expression (chi squared = 0.09, P = 0.764) and the groups with and without Gadd45a expression (chi squared = 0.14, P = 0.704). CONCLUSIONS: Both p53 and Gadd45a are highly expressed in human pancreatic cancer and may be associated with biological features of pancreatic cancer. Their expression alone or co-expression may be not helpful to evaluate the prognosis of patients with pancreatic cancer.