Patient-centered developments in colon- and rectal cancer with a multidisciplinary international team: From translational research to national guidelines.

Rarely, scientific developments centered around the patient as a whole are published. Our multidisciplinary group, headed by gastrointestinal surgeons, applied this research philosophy considering the most important aspects of the diseases "colon- and rectal cancer" in the long-term developments. Good expert cooperation/knowledge at the Comprehensive Cancer Center Ulm (CCCU) were applied in several phase III trials for multimodal treatments of primary tumors (MMT) and metastatic diseases (involving nearly 2000 patients and 64 centers), for treatment individualization of MMT and of metastatic disease, for psycho-oncology/quality of life involving the patients' wishes, and for disease prevention. Most of the targets initially were heavily rejected/discussed in the scientific communities, but now have become standards in treatments and national guidelines or are topics in modern translational research protocols involving molecular biology for e.g., "patient centered individualized treatment". In this context we also describe the paths we had to tread in order to realize our new goals, which at the end were highly beneficial for the patients from many points of view. This description is also important for students and young researchers who, with an actual view on our recent developments, might want to know how medical progress was achieved.

Low Pneumoperitoneum Pressure Improves Recovery of Transabdominal Preperitoneal Hernioplasty.

OBJECTIVE: To evaluate the recovery influence of CO2 pneumoperitoneum pressure for transabdominal preperitoneal hernioplasty (TAPP). STUDY DESIGN: Experimental study. PLACE AND DURATION OF STUDY: General Department II, Zhongda Hospital, Southeast University, Nanjing, China, from August 2016 to October 2018. METHODOLOGY: Eighty cases were enrolled prospectively and divided into three groups in chronological order. A 14 mmHg CO2 pressure was used for negative control group while the pressure was controlled at 12 mmHg for observation group and 10 mmHg for intervention group. General information included the patients' age, gender, type of hernia, hernia defect size, dissection of inguinal area, type of patch, time of operation, and frequency of swelling of perineum. Postoperative recovery was compared among the three groups at 24 hours and 1 month after surgery, including pain scores, foreign body sensation, local complications, urinary retention, swelling of the perineum, sex life and mobility. RESULTS: Seventy-eight patients were included in the final analysis. There were no differences among the three groups in patients' age, gender, type of hernia, hernia defect size, dissection of inguinal area and type of patch. However, the time of operation of intervention group increased (p=0.015) and incidence of swelling of perineum decreased than other two groups (p<0.05). After 24 hours, there were no significant differences in pain, foreign body sensation, local complications and urinary retention. Perineal swelling remission rate of intervention group was better than other two groups (p<0.05). After one month, three groups had no differences in the all terms of pain, foreign body sensation, sexual life and perineal swelling residual rate. CONCLUSION: Low pneumoperitoneum pressure can relieve swelling of perineum perioperatively and improve recovery of TAPP.

Bone morphogenetic protein-12 inducing tenogenic differentiation of mesenchymal stem cells enhances healing of linea alba incision.

The objective of this study was to investigate the curative effects of mesenchymal stem cells' tenogenic differentiation on linea alba incision healing induced by bone morphogenetic protein-12. Mesenchymal stem cells were isolated and induced by 10 ng/ml of bone morphogenetic protein-12 for 48 h. Expression of scleraxis, collagen I and collagen III were examined at 48 h, 5 and 7 days to investigate the tenogenic differentiation. The expression of scleraxis increases continually even in the absence of bone morphogenetic protein-12 for 5 days (P<0.01). The expression of collagen I and III requires persistent inducing. Then fifty Sprague-Dawley rats were randomly divided into five groups: negative control, positive control, sham group, native mesenchymal stem cells and tenogenically differentiated mesenchymal stem cells. Tensiometric testing and modified semiquantitative histological analysis were performed to explore the curative effects. The tension levels in the positive control, sham, native mesenchymal stem cells and tenogenically differentiated mesenchymal stem cells were 44, 41.8, 51.6 and 69.7%, respectively, compared with the negative control. Tenogenically differentiated mesenchymal stem cells exhibited a greater increase in tension compared with positive control, sham and native mesenchymal stem cell groups (P<0.05). From the sections stained with Masson's Trichrome, collagen organization and amount of tenogenically differentiated mesenchymal stem cells was better than the other three groups (P<0.05). In conclusion, mesenchymal stem cells' tenogenic differentiation induced by bone morphogenetic protein-12 can enhance linea alba incision healing.

Exosome: a novel mediator in drug resistance of cancer cells.

Exosomes are small membrane vesicles with a diameter of 40-100 nm, which are released into the intracellular environment. Exosomes could influence the genetic and epigenetic changes of receptor cells by promoting the horizontal transfer of various proteins or RNAs, especially miRNAs. Moreover, exosomes also play an important role in tumor microenvironment. Exosomes could promote the short- and long-distance exchanges of genetic information by acting as mediators of cell-to-cell communication. In addition, exosomes participate in drug resistance of tumor cells by genetic exchange between cells. It is reported that exosomes could be absorbed by recipient cells and transmit chemoresistance from drug-resistant tumor cells to sensitive ones. Then understanding the mechanisms of chemotherapy failure and controlling tumor progression effectively will be a major challenge for us. Therefore, in this review, we will briefly reveal the role of exosomes in drug resistance.

Circular RNA hsa_circ_0052112 promotes cell migration and invasion by acting as sponge for miR-125a-5p in breast cancer.

OBJECTIVES: Accumulating evidence has been reported that circular RNAs (circRNAs) are a class of relatively stable, non-coding RNAs, which are involved in the progression of many types of diseases. However, the mechanism of hsa_circ_0052112 in breast cancer cells is not entirely clear. Hsa_circ_0052112, generated from the ZNF83 gene, is selected by analyzing circRNA expression profiles of breast cancer cell by using microarray assay. In this study, we will show the role of hsa_circ_0052112 in regulating cell invasion and migration in breast cancer. METHODS: The expression level of hsa_circ_0052112 in MCF-7 and MDA-MB-231 cells was detected by RT-qPCR; we performed transwell assay to evaluate breast cancer cells' migration and invasion; predicated circRNA/miRNAs interaction using the miRanda and RNAhybrid software; identified the relationship between hsa_circ_0052112 and miR-125a-5p by luciferase activity assay and show the localization of hsa_circ_0052112 by FISH assay and show the significance of ZNF83 in clinical prognosis by Kaplan-Meier survival analysis. RESULTS: Hsa_circ_0052112 expression was significantly higher in MDA-MB-231 cells than that in MCF-7 cells. Overexpression of hsa_circ_0052112 promoted cell migration and invasion in breast cancer. Inversely, down-regulation of hsa_circ_0052112 suppressed breast cancer cells migration and invasion. Hsa_circ_0052112 was mostly located in cytoplasm. Hsa_circ_0052112 could directly sponge to miR-125a-5p; overexpression of miR-125a-5p significantly inhibited breast cancer cells migration and invasion. However, high or low expression of miR-125a-5p was not correlated with relapse free survival (RFS) by TCGA database validation, but high expression of ZNF83 was closely correlated with poor RFS by Kaplan-Meier plotter. CONCLUSIONS: These data suggest that hsa_circ_0052112 may be a potent biomarker for breast cancer, and may provide a new perspective on treatment of breast cancer.

Circular RNA hsa_circ_0072995 promotes breast cancer cell migration and invasion through sponge for miR-30c-2-3p.

AIM: To study the role of hsa_circ_0072995 in regulating the invasion and migration of breast cancer cells. MATERIALS & METHODS: Hsa_circ_0072995 expression was confirmed by quantitative real-time PCR; evaluating the migration and invasion of breast cancer cells through transwell assay; predicating circRNA/microRNAs interaction using the miRanda and RNAhybrid software; identifying the relationship between hsa_circ_0072995 and miR-30c-2-3p by luciferase activity assay; detecting the location of hsa_circ_0072995 by Fluorescence in situ hybridization assay. RESULTS: Hsa_circ_0072995 was significantly upregulated in MDA-MB-231 cells compared with MCF-7 cells. Hsa_circ_0072995 regulated the invasion and migration of breast cancer cells. Hsa_circ_0072995 existed in the nucleus and cytoplasm, and the proportion of the two was roughly equal. Hsa_circ_0072995 bound to miR-30c-2-3p. Overexpression of miR-30c-2-3p inhibited breast cancer cells migration and invasion. Low expression of miR-30c-2-3p was correlated with poor overall survival by The Cancer Genome Atlas database. CONCLUSION: Hsa_circ_0072995 may be a novel biomarker for breast cancer, and may function in metastasis of breast cancer.

CircRNA: a novel type of biomarker for cancer.

Circular RNAs (circRNAs) are a class of long, non-coding RNAs molecules that shape a covalently closed continuous loop which have no 5'-3' polarity and contain no polyA tail. CircRNAs also possess relatively jarless framework and are highly tissue-specific expressed in the eukaryotic transcriptome. Emerging evidences have discovered that thousands of endogenous circRNAs are present in mammalian cells and they mediate gene expression at the transcriptional or post-transcriptional level by binding to microRNAs or other molecules and then inhibit their function. Similarly, increasing evidence indicates that circRNAs may play a role in the development of several types of diseases, including atherosclerotic vascular disease risk, neurological disorders, prion diseases, osteoarthritis and diabetes. Furthermore, circRNAs exhibit aberrant expression in multiform types of cancer, including colorectal cancer, hepatocellular carcinoma and pancreatic ductal adenocarcinoma. And based on the function of circRNAs in cancer, we believe that circRNAs may serve as diagnostic or tumor promising biomarkers. Moreover, it will provide a new therapeutic target for the treatment of cancer.

Triptolide exerts protective effects against fibrosis following ileocolonic anastomosis by mechanisms involving the miR-16-1/HSP70 pathway in IL-10-deficient mice.

Surgeries, particularly ileocecal resection (ICR), are often required in the treatment of Crohn's disease (CD). However, recurrences are common for patients who undergo ICR, and anastomotic fibrosis is the main cause of re-operation. The present study aimed to investigate the therapeutic effects of triptolide (TPL) in ameliorating fibrosis following ileocolonic anastomosis. A model of IL­10­/­ mice undergoing ICR was used to study postsurgical inflammation and fibrosis of anastomosis. For this purpsose, interleukin (IL)­10­/­ mice were randomly divided into 3 groups as follows: the control group, the saline­treated group subjected to ICR (ST­ICR) and the TPL­treated group subjected to ICR (TT­ICR). Wild­type (WT) mice of matching ages were assigned to the WT group. The effects of TPL treatment on ileocolonic anastomosis were determined by histopathological evaluation, western blot analysis and ELISA. The analysis of the effects of TPL treatment on microRNA­16­1 (miR­16­1) and heat shock protein 70 (HSP70) expression was carried out by RT­qPCR and western blot analysis. Compared with the control group, significantly higher inflammation scores following anastomosis were observed in the ST­ICR group (P<0.05), although reversion was observed in the TT­ICR group, which was consistent with changes in the area of CD4+ cell infiltration. The elevated fibrosis scores and the overexpression of procollagen I and III in the ST­ICR group were all inhibited by TPL. With an increase in the severity of inflammation and fibrosis, the levels of IL­6, tumor necrosis factor­α (TNF­α) and transforming growth factor­ß1 (TGF­ß1) increased; however, a significant decrease in these levels was observed following treatment with TPL (P<0.05). The results of RT­qPCR revealed that the upregulated miR­16­1 levels in the ST­ICR group were significantly reduced by TPL. HSP70, which can be inhibited by miR-16-1, ameliorates anastomotic inflammation and fibrosis. Thus, the present study demonstrates that TPL exerts a protective effect against fibrosis following anastomosis in CD. The miR­16­1/HSP70 signaling pathway, which can be regulated by TPL, may thus represent a novel therapeutic option in CD that deserves further investigation.

The role of miR-130a in cancer.

MicroRNAs (miRs) are short and highly conserved non-coding RNAs molecules consisting of 18-25 nucleotides that regulate gene expression at post-transcriptional level by direct binding to complementary binding sites within the 3'untranslated region (3'UTR) of target mRNAs. New evidences have demonstrated that miRNAs play an important role in diverse physiological processes, including regulating cell growth, apoptosis, metastasis, drug resistance, and invasion. In chromosomes 11 and 22 of the miR-130 family, paralogous miRNA sequences, miR-130a and miR-130b are situated, respectively. MiR-130a has participated in different pathogenesis, including hepatocellular carcinoma, cervical cancer, ovarian cancer, glioblastoma, prostate carcinoma, leukemia, etc. Most important of all, more and more evidences indicate that miR-130a is associated with drug resistance and acts as an intermediate in PI3 K/Akt/PTEN/mTOR, Wnt/ß-catenin and NF-kB/PTEN drug resistance signaling pathways. Drug resistance has emerged as a major obstacle to successful treatment of cancer nowadays and in this review, we will reveal the function of miR-130a in cancer, especially in drug resistance. Therefore, it will provide a new therapeutic target for the treatment of cancer, especially in chemotherapy.

Comparison of different sets of instruments for laparoendoscopic single-site surgery in a surgical simulator with novices.

BACKGROUND: Given the parallel entry of working instruments through a single incision in laparoendoscopic single-site surgery, loss of triangulation in the abdominal cavity and counteracting movements of the instruments are inevitable obstacles. Some specially designed devices have emerged to ameliorate these challenges. METHODS: Twenty-four novice participants were randomized into four groups using assigned instruments, conventional straight instruments, single-curved instruments, double-curved instruments and articulating instruments, respectively, to perform two basic tasks (peg transferring and pattern cutting) 14 times in a modified simulator. A test of the tasks and a resection of the intestine segment of a rat were performed. The task scores and evaluation of intraoperative skills during the resection of the intestine segment were recorded. The instrument of modified National Aeronautics and Space Administration Task Load Index (NASA-TLX) was completed. RESULTS: The task scores of the groups using single-curved instruments and articulating instruments were better than the other two groups on the simulator tasks, consistent with the evaluation of intraoperative skills during the resection of intestine segment. As the proficiency with the instruments increased, the task scores improved, as demonstrated by the learning curve. The workload measured by the modified NASA-TLX tool demonstrated that the groups using articulating instruments and double-curved instruments had a heavier workload in most of the categories compared with the other two groups. CONCLUSIONS: Single-curved and articulating instruments are more effective than conventional straight and double-curved devices, and are favourable in laparoendoscopic single-site surgery for novice learners.

Pioglitazone reduces lipid droplets in cholesterolosis of the gallbladder by increasing ABCA1 and NCEH1 expression.

As a cholesterol-induced metabolic disease, cholesterolosis of the gallbladder is often resected clinically, which could lead to many complications. The histopathology of cholesterolosis is due to excessive lipid droplet accumulation in epithelial and subcutaneous tissues. The main components of lipid droplets are cholesterol esters (CEs). Removal of CEs from gallbladder epithelial cells (GBECs) is very important for maintaining intracellular cholesterol homeostasis and for treating cholesterol-related diseases. In this study, pioglitazone was used to reduce intracellular CEs. To further elucidate the mechanism, cholesterolosis GBECs were treated with pioglitazone, 22-(R)-hydroxycholesterol (a liver X receptor α (LXRα) agonist), or peroxisome proliferator-activated receptor gamma (PPARγ) siRNA. Western blotting for PPARγ, LXRα, ATP-binding cassette transporter A1 (ABCA1), and neutral cholesteryl ester hydrolase 1 (NCEH1) was performed. At length, cholesterol efflux to apoA-I was measured, and oil red O staining was used to visualize lipid droplet variations in cells. In conclusion, we observed that pioglitazone increased ABCA1 expression in an LXR-dependent manner and NCEH1 expression in an LXRα-independent manner, which mobilized CE hydrolysis and cholesterol efflux to reduce lipid droplet content in cholesterolosis GBECs. Our data provide a plausible alternative to human gallbladder cholesterolosis.

Laparoendoscopic single-site distal pancreatectomy in pigs.

AIM: To explore the technique for laparoendoscopic single-site distal pancreatectomy. METHODS: Laparoendoscopic single-site spleen-preserving distal pancreatectomy was performed in pigs using a novel flexible multichannel port, a curved laparoscopic multifunctional operative device and a fish hook retractor, which provided a favorable operative field. RESULTS: Six pigs were involved in this study, and five survived the procedure. The first animal died following injury to the superior mesenteric vein and uncontrolled intraoperative bleeding. Except for this failure, the mean operative time was 155 min (range: 102-236 min). A steep learning curve was observed in the study, with a mean operative time of 177 min in the first two operations vs 134 min in the last three operations. The mean blood loss was 50 mL, and the postoperative course was uneventful. The animals were sacrificed three weeks after the procedures, and no pancreatic leakage or abdominal infection was found macroscopically. CONCLUSION: Laparoendoscopic single-site distal pancreatectomy is a safe and feasible procedure and can be implemented in humans in selected cases at qualified surgical centers.

22(R)-hydroxycholesterol and pioglitazone synergistically decrease cholesterol ester via the PPARγ-LXRα-ABCA1 pathway in cholesterosis of the gallbladder.

Cholesterosis is a disease of cholesterol metabolism characterized by the presence of excessive lipid droplets in the cytoplasm. These lipid droplets are mainly composed of cholesterol esters derived from free cholesterol. The removal of excess cholesterol from gallbladder epithelial cells (GBECs) is very important for the maintenance of intracellular cholesterol homeostasis and the preservation of gallbladder function. Several lines of evidence have indicated that the activation of either peroxisome proliferator-activated receptor gamma (PPARγ) or liver X receptor α (LXRα) relates to cholesterol efflux. While pioglitazone can regulate the activation of PPARγ, 22(R)-hydroxycholesterol can activate LXRα and is a metabolic intermediate in the biosynthesis of steroid hormones. However, the effect of 22(R)-hydroxycholesterol in combination with pioglitazone on cholesterosis of the gallbladder is unclear. GBECs were treated with pioglitazone, 22(R)-hydroxycholesterol or PPARγ siRNA followed by Western blot analysis for ATP-binding cassette transporter A1 (ABCA1), PPARγ and LXRα. Cholesterol efflux to apoA-I was determined, and Oil Red O staining was performed to monitor variations in lipid levels in treated GBECs. Our data showed that 22(R)-hydroxycholesterol can modestly up-regulate LXRα while simultaneously increasing ABCA1 by 56%. The combination of 22(R)-hydroxycholesterol and pioglitazone resulted in a 3.64-fold increase in ABCA1 expression and a high rate of cholesterol efflux. Oil Red O staining showed an obvious reduction in the lipid droplets associated with cholesterosis in GBECs. In conclusion, the present findings indicate that the anti-lipid deposition action of 22(R)-hydroxycholesterol combined with pioglitazone involves the activation of the PPARγ-LXRα-ABCA1 pathway, increased ABCA1 expression and the efflux of cholesterol from GBECs. Thus, 22(R)-hydroxycholesterol synergistically combined with pioglitazone to produce a remarkable effect on lipid deposition in cholesterosis GBECs.

Decreased SCF/c-kit signaling pathway contributes to loss of interstitial cells of Cajal in gallstone disease.

Cholecystolithiasis is a common disease, and gallbladder dysmotility is considered as a pivotal pathogenesis. Interstitial cells of Cajal (ICCs) serve as pacemakers and mediators of neuromuscular transmission for gastrointestinal motility. Reduction of ICCs has been reported in gallstone diseases. However, there are no reasonable mechanisms for the cholecystolithiasis-associated loss of ICCs in humans. Stem cell factor (SCF) and its ligand c-kit are essential for normal development and survival of ICCs. To date, little is known about the SCF/c-kit signaling pathway in gallstone diseases. The purpose of this study was to investigate the role of the SCF/c-kit signaling pathway in the loss of ICCs in cholecystolithiasis. Data from 18 patients with gallstones and 14 individuals without gallstones were compared. The gallbladder contractility was assessed by measuring the gallbladder ejection fraction (GEF) ultrasonographically. Tissues samples were obtained during surgery, changes of ICC quantities were analyzed by immunohistochemistry, and the mRNA and protein expression of SCF and c-kit were detected by Real-Time PCR and Western-blot analysis. Compared with the controls, the GEF was significantly reduced in the gallstone group, and decreased number of ICCs was present obviously in the gallstone group. Furthermore, the mRNA and protein expression of SCF and c-kit were significantly attenuated in the gallstone group. These data indicate that gallbladder motility may be affected by reduction of ICCs in gallstone disease. Additionally, the decreased of SCF/c-kit signaling pathway play an important role in the loss of ICCs.

An investigation for public acceptance of laparoendoscopic single-site surgery.

UNLABELLED: Objective : Laparoendoscopic single-site surgery (LESS) is the latest innovation in minimally invasive surgery with unconfirmed advantages. The public perception of LESS is the basis of carrying out the surgery. METHODOLOGY: Participants from the outpatient department were invited to rate, on a 5-point Likert scale, the important factors including scar, complications, cost, pain and hospital stay in choosing surgery. In addition, those who preferred LESS would continue to make their choices as the risks of LESS in above mentioned aspects rose. RESULTS: About 85% of the questionnaires were included in the analysis. Complication was the most important factor with an average score of 4.77±0.43, followed by pain (3.84±0.96), scar (3.57±1.17), cost (3.41±0.87) and hospital stay (3.04±0.86). Of the 196 participants, 132 (67%) preferred LESS with younger age (35.3±10.64 versus 40.4 ±9.6, P=0.001). Better cosmesis was the only factor that made the participants choose LESS (3.78±1.11 versus 3.13±1.19, P<0.005). Almost 90% of the participants could accept the hypothesis (incision length of 3.5cm, cost up to 120%, pain up to 120%, hospital stay of 5 days), while only 50% of participants could accept the risk of complications of 6%. CONCLUSIONS: Complication is the most important factor that the public are concerned about in choosing surgery. LESS is preferred by young who care more concerned about the cosmesis, even with moderately elevated risks of extending incision and increasing hospital cost, postoperative pain and hospital stay.

Transumbilical single-incision laparoscopic appendectomy using conventional instruments: the single working channel technique.

BACKGROUND: This study aimed to evaluate the feasibility, safety, and cosmetic results of a novel technique, transumbilical single-incision laparoscopic appendectomy (TSILA), using a single working channel with conventional instruments. PATIENTS AND METHODS: The study enrolled 84 consecutive patients undergoing laparoscopic appendectomy for acute appendicitis. To test the advantages of TSILA on the management of patients with acute appendicitis, a prospective randomized clinical trial was conducted. Surgical outcomes such as operation time, complication, and hospital stay of 42 patients undergoing TSILA were analyzed and compared with those of 42 patients undergoing classic 3-port appendectomy. All patients received a follow-up visit for 3 to 12 months. RESULTS: The study consisted of 42 patients undergoing TSILA and 42 patients undergoing classic 3-port laparoscopic appendectomy with an average age of 34.1 and 34.9 years, respectively. The mean operative time of TSILA did not show any difference when compared with the classic procedure (84.8 vs. 77.9 min, P=0.271). No operative complications occurred in patients undergoing TSILA, whereas 2 patients undergoing the classic procedure showed incisional infection. The average postoperative hospital stay was 2.7 days in the TSILA group and 2.9 days in the classic procedure group with no difference (P=0.316). At the follow-up visit, no patient showed any evidence of incisional hernia. The transumbilical incisions were visible minimally, and the cosmetic scores given by patients undergoing TSILA was higher than that given by patients undergoing the classic procedure (4.5 vs. 3.9, P<0.001). CONCLUSIONS: The results of the study demonstrate that laparoscopic appendectomy can be achieved through a single umbilical incision and a single working channel using conventional instruments and that this approach is successful, safe, economic, and esthetic.

A new strategy of minimally invasive surgery for cholecystolithiasis: calculi removal and gallbladder preservation.

BACKGROUND: Cholecystolithiasis is a common disease. Cholecystectomy is the main treatment method but is associated with various complications in some patients. This study explores a novel, minimally invasive surgery for the removal of calculi and the preservation of the gallbladder using a laparoscope combined with the soft choledochoscope. METHOD: A retrospective analysis was conducted between January 2010 and December 2012 in 65 patients with cholecystolithiasis who underwent the minimally invasive surgery for calculi removal and gallbladder preservation. RESULTS: In 61 cases of gallstone removal, the gallbladder was preserved perfectly with no complications. The other 4 cases were switched to laparoscopic cholecystectomy because of tiny stones blocking the cystic duct or submucosal stones. The success rate was 93.8%. Follow-up included both clinical assessment and ultrasound examination every 6 months after the operation. The patients with preoperative symptoms were symptom-free, and gallbladder function was well preserved. The overall stone recurrence rate was 4.92% at a mean follow-up of 26 months (range 6-40). CONCLUSIONS: Using the laparoscope combined with the soft choledochoscope for gallbladder-preserving cholecystolithotomy can remove stones, preserve gallbladder function, and effectively avoid the various complications of cholecystectomy. In our follow-up, gallbladder function was not affected and the stone recurrence rate was quite low.

Laparoendoscopic single-site cholecystectomy versus conventional laparoscopic cholecystectomy: a systematic review of randomized controlled trials.

BACKGROUND: Laparoendoscopic single-site surgery is a revolution in minimally invasive surgery in recent years, and cholecystectomy is the most common intervention. However, laparoendoscopic single-site cholecystectomy (LESSC) is a controversial procedure. Its safety, cosmetic results, complications, post-operative pain and post-operative stay are not confirmed by multi-centre randomized controlled studies. OBJECTIVES: This review examined the impact of LESSC versus conventional laparoscopic cholecystectomy (CLC). The primary outcomes were conversional rate and intraoperative and post-operative complications, and other outcomes were cosmetic results, operative time, post-operative pain and post-operative stay. DATA SOURCES: Pubmed, EMbase, Web of Science, Ovid, clinical trials of the US National Institutes of Health and Cochrane Database of Systematic Reviews and Controlled Trials Register were searched to identify all possible randomized controlled trials (RCTs). METHODS: Two reviewers completed article search, and eligible data were entered into a computerized spreadsheet for analysis. RESULTS: Nine studies were obtained using the search strategy. Patients accepting LESSC had a higher conversion rate than those having CLC (7.17 (3.00, 17.11) (P < 0.01)). There was no significant difference for total complications between the two groups (1.17 (0.76, 1.80) (P = 0.46)), but incidence of incisional hernia and haemorrhage (seroma) in the LESSC group was obviously higher than that in the CLC group. Operative time was much longer in the LESSC group (mean difference: 10.69 (3.14, 18.24) (P = 0.006)). Cosmetic results favoured LESSC; however, post-operative abdominal pain and post-operative stay were not significantly different between the two groups in most of the nine studies. CONCLUSIONS: LESSC can be safely performed with better cosmetic results; however, it carries a higher conversion rate and a longer operative time, and offers no benefit in terms of post-operative abdominal pain and post-operative stay. Further RCTs, according to the CONSORT statement, are advocated to validate its objective benefits.

Cyclopamine reverts acquired chemoresistance and down-regulates cancer stem cell markers in pancreatic cancer cell lines.

BACKGROUND: The hedgehog (Hh) pathway has been implicated in the pathogenesis of cancer including pancreatic ductal adenocarcinoma (PDAC). Recent studies have suggested that Hh plays an important role in maintaining the cancer stem cell (CSCs) pool. Gemcitabine-resistant pancreatic cancer cells highly express some of the CSCs markers. However, the expression level of Hh members in gemcitabine-resistant pancreatic cancer cells remains unknown. The aim of this study was to verify the expression of HH members, such as Shh, Ptc, SMO and Gli-1 in gemcitabine-resistant PDAC cell lines, and to explore a new strategy to overcome chemoresistance in PDAC. MATERIAL AND METHODS: Quantitative real-time RT-PCR (Q-PCR) and western blot were used to evaluate the relative expression level of HH members in SW1990, CFPAC-1 cells and gemcitabine-resistant SW1990, CFPAC-1 cells. The change of cancer stem cell markers and the expression level of HH members before and after cyclopamine treatment was evaluated using flow cytometry and Q-PCR, western blot, respectively. Cell apoptosis after cyclopamine treatment was measured by flow cytometry. RESULTS: CD44, CD133 and the expression level of HH members, including Shh, SMO, Gli-1, were found to be highly expressed in gemcitabine-resistant cells, which were significantly down-regulated by cyclopamine treatment. Flow cytometry analysis showed increased cell apoptosis after cyclopamine treatment. CONCLUSION: Gemcitabine-resistant pancreatic cancer cells highly express CSCs markers and some of the HH members, and inhibition of HH by cyclopamine is an effective method of reversing gemcitabine resistance in pancreatic cancer.

Side population in the pancreatic cancer cell lines SW1990 and CFPAC-1 is enriched with cancer stem-like cells.

In this study, we first sought to determine the existence of side population (SP) cells in pancreatic cancer cell lines. Furthermore, we compared the biological characteristics of SP and non-SP cells. The presence of side population cells in pancreatic cancer cell lines was detected by Hoechst 33342 staining and FACS analysis. Cell cycle distribution was analyzed using flow cytometry. SP and non-SP cells were exposed to various concentrations of gemcitabine; drug sensitivity was examined using the MTT assay and flow cytometry using Annexin-V and PI staining. To compare the tumorigenic ability in vivo, groups of nude mice were orthotopically inoculated with varying numbers of SP and non-SP cells. The percentages of CD44+CD24+ and CD133+ in SP and non-SP cells were also detected by FACS analysis. The SP fraction was detected in BxPc-3, CFPAC-1, MIA PaCa-2, PANC-1 and SW1990 pancreatic cancer cell lines. Cell cycle analysis revealed that the SP cells contained more cells in the G1 phase and fewer cells in the S phase when compared with the non-SP cells. The SP cells exhibited increased tumorigenetic ability following in vivo transplantation into BALB/C nude mice and increased chemoresistance following in vitro exposure to gemcitabine. FACS analysis showed that the SP cells contained more CD44+CD24+ and CD133+ cells than the non-SP cells. In conclusion, these observations suggest that SP cells in the pancreatic cancer cell lines possess the property of cancer stem cells. SP cells may therefore be novel specific targets for the effective treatment of pancreatic cancer.