Pathway-guided analysis identifies Myc-dependent alternative pre-mRNA splicing in aggressive prostate cancers.

We sought to define the landscape of alternative pre-mRNA splicing in prostate cancers and the relationship of exon choice to known cancer driver alterations. To do so, we compiled a metadataset composed of 876 RNA-sequencing (RNA-Seq) samples from five publicly available sources representing a range of prostate phenotypes from normal tissue to drug-resistant metastases. We subjected these samples to exon-level analysis with rMATS-turbo, purpose-built software designed for large-scale analyses of splicing, and identified 13,149 high-confidence cassette exon events with variable incorporation across samples. We then developed a computational framework, pathway enrichment-guided activity study of alternative splicing (PEGASAS), to correlate transcriptional signatures of 50 different cancer driver pathways with these alternative splicing events. We discovered that Myc signaling was correlated with incorporation of a set of 1,039 cassette exons enriched in genes encoding RNA binding proteins. Using a human prostate epithelial transformation assay, we confirmed the Myc regulation of 147 of these exons, many of which introduced frameshifts or encoded premature stop codons. Our results connect changes in alternative pre-mRNA splicing to oncogenic alterations common in prostate and many other cancers. We also establish a role for Myc in regulating RNA splicing by controlling the incorporation of nonsense-mediated decay-determinant exons in genes encoding RNA binding proteins.

Resveratrol inhibits IL-1ß-mediated nucleus pulposus cell apoptosis through regulating the PI3K/Akt pathway.

Nucleus pulposus (NP) cell apoptosis is a classical cellular character during intervertebral disc degeneration (IDD). Previous studies have shown that inflammatory cytokine-induced NP cell apoptosis plays an important role in disc degeneration. The present study was aimed to investigate whether resveratrol can suppress IL-1ß-mediated NP cell apoptosis and the potential signal transduction pathway. Experimental rat NP cells were treated with culture medium containing IL-1ß (20 ng/ml) for 7 days. Control NP cells were cultured in the baseline medium. Resveratrol was added along with culture medium to investigate its effects. The inhibitor LY294002 was used to study the role of the PI3K/Akt pathway. NP cell apoptosis was reflected by the caspase-3 activity, cell apoptosis ratio, and expression of apoptosis-related molecules (Bcl-2, Bax, caspase-3, cleaved caspase-3, and cleaved PARP). Compared with the control NP cells, IL-1ß significantly increased caspase-3 activity, NP cell apoptosis ratio and mRNA/protein expression of Bax, caspase-3, cleaved caspase-3 and cleaved PARP, but decreased mRNA expression of Bcl-2. However, resveratrol partly suppressed the effects of IL-1ß on those cell apoptosis-related parameters. Further analysis showed that IL-1ß significantly decreased activity of the PI3K/Akt pathway whereas resveratrol partly increased activity of the PI3K/Akt pathway in NP cells treated with IL-1ß. Additionally, when the inhibitor LY294002 was added along with the resveratrol, its protective effects against IL-1ß-induced NP cell apoptosis were attenuated. In conclusion, resveratrol suppresses IL-1ß-mediated NP cell apoptosis through activating the PI3K/Akt pathway. Resveratrol may be an effective drug to attenuate inflammatory cytokine-induced disc degenerative changes.

Deep-learning augmented RNA-seq analysis of transcript splicing.

A major limitation of RNA sequencing (RNA-seq) analysis of alternative splicing is its reliance on high sequencing coverage. We report DARTS (https://github.com/Xinglab/DARTS), a computational framework that integrates deep-learning-based predictions with empirical RNA-seq evidence to infer differential alternative splicing between biological samples. DARTS leverages public RNA-seq big data to provide a knowledge base of splicing regulation via deep learning, thereby helping researchers better characterize alternative splicing using RNA-seq datasets even with modest coverage.

Laparoscopic resection of benign schwannoma in the hepatoduodenal ligament: A case report and review of the literature.

Schwannomas are mesenchymal neoplasms with a low malignant potential, which arise from Schwann cells. The tumors can occur in most parts of the body; however, the head, neck and flexor surfaces of the extremities are the most common locations. Schwannomas occurring in the hepatoduodenal ligament are extremely rare. To the best of our knowledge, only two cases of schwannoma in the hepatoduodenal ligament have been reported in the literature, and treatment of such cases by laparoscopic surgery has not yet been reported. The present study reports a case of schwannoma in the hepatoduodenal ligament in a 50-year-old male patient. Physical and laboratory examinations showed no abnormal results. Ultrasound and computed tomography failed to definitively diagnose the mass and identify its location. During laparoscopic surgery, a mass was identified in the hepatoduodenal ligament and was completely removed. The gross specimen was a 4.5×2.5×2.5-cm localized mass, yellowish-white in color. Microscopic examination revealed that the tumor was mainly composed of spindle-shaped cells and no atypical cells were identified. Immunohistochemical staining showed a strong positive S-100 protein reaction, whereas cluster of differentiation 34 and epithelial membrane antigen were negative. The final diagnosis of the lesion was benign schwannoma of the hepatoduodenal ligament. The patient was followed-up for 7 months and, at the time of writing, was healthy and without any complications. The aim of the present study was to describe a rare case of hepatoduodenal ligament schwannoma in a 50-year-old male patient, and present a review of the literature. To the best of our knowledge, this is the first case of hepatoduodenal ligament schwannoma treated by laparoscopic surgery.

Comparison of two laparoscopic splenectomy plus pericardial devascularization techniques for management of portal hypertension and hypersplenism.

OBJECTIVE: Our research was conducted to analyze the outcomes of two laparoscopic splenectomy plus pericardial devascularization (LSPD) techniques in the management of portal hypertension (PTH) and hypersplenism. METHODS: Between May 2012 and May 2013, 41 patients with PTH and hypersplenism undergoing LSPD were retrospectively analyzed. Of them, 29 patients received LSPD by LigaSure Vessel Sealing System (LVSS) and Endo-GIA universal endoscopic vascular linear staplers (Endo-GIA) (EG Group) and 12 patients received LSPD by LVSS and Hem-o-Lock (HL Group). Operating time, intraoperative blood loss, postoperative course, and hospitalization costs were compared between the two LSPD combination techniques. RESULTS: There were no significant differences in preoperative patient characteristics of the two groups. Significantly less operating time, intraoperative blood loss, and postoperative complications were observed in EG Group. The incidence of portal vein thrombosis was lower in the EG Group (3.4 vs. 8.3%), as well as the incidence of pancreatic fistula (0 vs. 8.3%). Upper gastrointestinal hemorrhage was not observed in either group. Uncontrolled bleeding warranted conversion to open surgery in one case in EG Group (conversion rate 3.4%) and in two cases in HL Group (conversion rate 16.7%). Two patients (16.7%) in HL Group underwent successful emergency exploratory laparotomy due to uncontrolled intraabdominal bleeding postoperatively. No re-operation was needed in EG Group. Two patients experienced liver failure after surgery in each group. Of those, three patients were managed successfully and one patient refused further therapy. While the overall complication rate was significantly lower in EG Group (17.2 vs. 58.3%, P < 0.05), overall hospitalization costs remained significantly higher for EG Group. CONCLUSION: The results suggest that the modified Endo-GIA and LVSS technique is a safe and effective combination approach to LSPD with shorter operative time, less intraoperative blood loss, lower conversion rate to laparotomy, shorter hospital stay, better recovery, and lower postoperative complication rate compared with the Hem-o-Lock and LVSS approach. Higher hospitalization expenses associated with the Endo-GIA and LVSS approach.

Effect of Yinxieling decoction on PASI, TNF-α and IL-8 in patients with psoriasis vulgaris.

OBJECTIVE: To study the effect of Yinxieling decoction on PASI, TNF-α and IL-8 in patients with psoriasis vulgaris. METHODS: A total of 120 cases of psoriasis vulgaris were divided into 4 groups according to syndrome differentiation of TCM and randomized controlled method: wind heat syndrome group (group A), blood stasis syndrome group (group B), blood dryness syndrome group (group C) and control group (group D) (n=30 per group). Patients in observation groups were treated with Yinxieling decoction, while patients in control group were treated by placebo for 8 weeks. Levels of TNF-α and IL-8 were determined before treatment, 4 and 8 weeks after treatment. psoriasis area and severity index score was also performed before and after treatment. RESULTS: psoriasis area and severity index score and serum level of TNF-α, IL-8 were significantly decreased in all groups. The decrease in three observation groups was more significant (P<0.05 or P<0.01), and the decrease in wind heat syndrome group was the most significant (P<0.01). psoriasis area and severity index was positively correlated with TNF-α and IL-8, respectively (P<0.05). CONCLUSIONS: Yinxieling decoction has therapeutical effect on psoriasis vulgaris via regulating TNF-α and IL-8.

[Efficacy analysis of laparoscopic sleeve gastrectomy in the treatment of obesity-related comorbidities].

OBJECTIVE: To investigate the safety and efficacy of laparoscopic sleeve gastrectomy (LSG) for the treatment of obesity and related diseases. METHODS: Clinical data of 67 patients who underwent LSG between December 2006 and July 2011 were analyzed retrospectively. Improvements in body mass index(BMI), percentage of excess weight loss(EWL), type 2 diabetes mellitus, hypertension and other comorbidities were observed at one year postoperatively. RESULTS: Laparoscopic procedures were completed in 67 patients without conversion. The operative time was(78±17) min. The postoperative hospital stay was(5.0±1.7) d. The postoperative recovery was uneventful and there were no perioperative death or severe postoperative complication. Sixty-four patients(95.5%) had a postoperative follow up of 1 year. One year after LSG, BMI decreased by(10.4±3.7) kg/m(2) from (37.7±4.1) kg/m(2) preoperatively and EWL was(80.2±27.7)%. In 13 cases of type 2 diabetes mellitus, 7 patients (53.8%, 7/13) were weaned off hypoglycemic agents or insulin. In 11 cases of hypertension, 5 required no medications(45.5%, 5/11). The remission rate was both 100%. There was significant resolution or improvement of other obesity-related comorbidiities, including hyper-triglyceridemia(n=51), hyperuricemia(n=42), sleep apnea syndrome (n=2), osteoarticular disease (n=9), and acanthosis (n=8). CONCLUSIONS: LSG is safe and feasible for the treatment of obesity and can cure or improve type 2 diabetes mellitus, hypertension and other obesity related comorbidities.

Laparoscopic splenectomy and azygoportal disconnection with intraoperative splenic blood salvage.

BACKGROUND: Intraoperative blood salvage can reduce or avoid perioperative allogeneic blood transfusion. Salvaging the blood in the portal hypertension-induced enlarged spleen becomes an issue of concern during devascularization surgery because an enlarged spleen accommodates a large red cell pool. We report 20 cases of laparoscopic splenectomy and azygoportal disconnection and present the advantages of the use of intraoperative splenic blood salvage during the procedure. METHODS: A total of 20 cirrhotic patients with esophagogastric variceal bleeding refractory to treatment with ß-blockers and endoscopic therapy were studied. Laparoscopic splenectomy with azygoportal disconnection was performed. During the procedure, an intraoperative autologous blood salvage device recovered the splenic blood. The perioperative data were recorded from various viewpoints. RESULTS: The operative time was 3.1 ± 0.3 h and the blood loss was 70.5 ± 32.5 ml. The weight of the excised and morcellated spleen was 826.0 ± 155.1 g. The volume of autotransfused blood was 541.0 ± 150.4 ml. No patient received a perioperative allogeneic blood transfusion. There were no significant complications either intraoperatively or postoperatively. The hemoglobin value increased from 9.3 ± 0.8 to 11.5 ± 1.1 g/dl at postoperative day 1 (p < 0.01). During a postoperative follow-up period of 18.0 ± 9.0 months for 18 patients, neither esophageal variceal bleeding nor encephalopathy recurred. CONCLUSION: Laparoscopic splenectomy with azygoportal disconnection is a feasible, effective, and safe surgical method for the treatment of bleeding portal hypertension. Intraoperative splenic blood salvage can avoid the risk associated with allogeneic transfusion during the procedure, with an advantage of significantly increased postoperative hemoglobin levels.

Comparative study of laparoscopic sleeve gastrectomy with and without partial enterectomy and omentectomy.

BACKGROUND: Laparoscopic sleeve gastrectomy (LSG) is a novel bariatric surgical procedure that constitutes the first-stage procedure of laparoscopic Roux-en-Y gastric bypass in high-risk patients, the long-term results of which are unknown. Our objective was to establish whether partial enterectomy and omentectomy are necessary in addition to LSG to achieve weight loss in obese patients. The setting was a case series in a provincial hospital. METHODS: A total of 40 obese patients (29 women and 11 men) were separated into 2 equal groups according to patient choice. Group 1 underwent LSG alone, and group 2 underwent LSG plus partial enterectomy and omentectomy. The partial enterectomy left the first 100 cm of the jejunum and the last 200 cm of the ileum. The data were collected during the follow-up examinations, performed at 1, 3, 6, and 12 months postoperatively. RESULTS: The body mass index loss (BMIL) was 3.9 ± .5 kg/m(2) and 9.4 ± 1.3 kg/m(2) at 1 and 12 months in group 1, respectively. The BMIL was 4.5 ± .9 kg/m(2) and 10.4 ± 1.9 kg/m(2) at 1 and 12 months in group 2, respectively. At 1 and 12 months postoperatively, the percentage of excess body weight loss was 32.2% ± 12.6% and 81.5% ± 20.4% in group 1 and 35.5% ± 10.5% and 83.8% ± 24.5% in group 2, respectively. Except for the BMIL at 1 month after surgery, no significant differences were found in the BMIL or percentage of excess body weight loss. CONCLUSION: LSG with and without partial enterectomy and omentectomy in our study was an effective method of bariatric surgery, but they did not differ in their effect on weight loss. However, the long-term effect of weight loss with LSG alone or combined with partial enterectomy and omentectomy needs additional study.

[Expression of Oct-4 transcription factor in squamous cell skin carcinoma and seborrheic keratosis].

OBJECTIVE: To detect the expression of stem cell transcription factor Oct-4 in squamous cell carcinoma (SCC) and seborrheic keratosis (SK) and its association with cancer stem cells. METHODS: Immunohistochemistry was used to detect Oct-4 expression in 35 SCC cases, 21 SK cases and 15 normal control skin tissues. RESULTS: Oct-4 expression was negative in normal skin and showed a significant difference between SCC and SK tissues (P<0.05). CONCLUSION: The Oct-4-positive cells in SCC and SK are probably tumor stem cells. Oct-4 expression may provide an important evidence for isolation and identification of human SCC and SK stem cells.

Laparoscopic versus open pediatric splenectomy for massive splenomegaly.

Laparoscopic splenectomy (LS) has rapidly evolved into the technique of choice compared with open splenectomy (OS) because of the advantages of the minimally invasive approach. Splenomegaly increases the technical difficulties of LS. LS for massive splenomegaly has generally been found to fail in adults and children. With improvements in laparoscopic technique and accumulation of laparoscopic experience, however, concerns about completing the procedure in pediatric cases with even massive splenomegaly have been lowered. A retrospective review (April 1997-October 2009) of databases at 2 institutions identified 145 children undergoing splenectomy, 79 laparoscopic and 66 open. We defined splenic margin below the umbilicus or anteriorly extending over the midline as massive splenomegaly. By this definition, 22 cases of pediatric laparoscopic and 17 cases of open splenectomies for massive splenomegaly were performed. Perioperative and follow-up data of laparoscopic pediatric splenectomies were compared with those of open splenectomies, including operative time, bleeding, spleen size, complications, and hospital stay. There were no deaths, wound infections, or instances of pancreatitis. No accessory spleen was missed by laparoscopic; accessory spleens were missed in 2 patients in open splenectomies. The complication rate of laparoscopic versus open was 13.6% versus 41.2%. No subsequent surgery was necessary for dealing with complications both in laparoscopic and open series. Laparoscopic pediatric splenectomy for massive splenomegaly is a feasible, effective, and safe procedure and is associated with low morbidity and a short hospital stay.

Laparoscopic splenectomy in portal hypertension: a single-surgeon 13-year experience.

BACKGROUND: Although laparoscopic splenectomy (LS) has become the standard approach for most splenectomy cases, some areas still remain controversial. To date, the indications that preclude LS are not clearly defined. Portal hypertension from liver cirrhosis still is a contraindication to LS in the clinical practice guidelines of the European Association for Endoscopic Surgery published in 2008. This study aimed to evaluate the feasibility of LS for hypersplenism secondary to liver cirrhosis and portal hypertension. METHODS: The study retrospectively analyzed 206 laparoscopic splenectomies performed for a variety of indications over 13 years. According to diagnosis, the patients were divided into group A (hypersplenism secondary to liver cirrhosis and portal hypertension, n = 96) and group B (hematologic and other disorders, n = 110). A detailed review of medical records was conducted. The perioperative data for the two groups were compared including patient characteristics, diagnosis, operative details, complication rates, and postoperative hospital stay. RESULTS: Laparoscopic splenectomy was completed for 201 patients. Conversion from laparoscopic to open surgery was necessary for 5 patients (2.4%) because of hemorrhage, and 26 patients (12.6%) had complications. There were significant differences between groups A and B in terms of mean operation time (2.8 vs. 2.1 h), complication rates (17.7% vs. 8.2%), and postoperative stay (7.1 vs. 4.7 days). However, the two groups showed no significant differences with respect to intraoperative blood loss, blood transfusion, and conversion rate. CONCLUSION: Laparoscopic splenectomy is a feasible, effective, and safe surgical procedure for patients who require splenectomy. Hypersplenism secondary to cirrhosis and portal hypertension should not be considered contraindications for LS.

Laparoscopic versus open splenectomy for hypersplenism secondary to liver cirrhosis.

BACKGROUND: Since the first laparoscopic splenectomy (LS) in 1991, LS has become the gold standard for the removal of normal-to-moderately enlarged spleens in benign conditions. Compared with open splenectomy (OS), fewer postsurgical complications and better postoperative recovery were observed, but it is contraindicated for hypersplenism secondary to liver cirrhosis owing to technical difficulties associated with splenomegaly, well-developed collateral circulation, and increased risk of bleeding. With the improvements of laparoscopic technique, the concept is changing. METHODS: OS and LS performed for hypersplenism secondary to liver cirrhosis at our institution were analyzed. Relationships between postoperative increases in platelet counts, white blood cell counts, hemoglobin, and liver function were examined. Perioperative data of LSs were compared with those of OSs, including operative time, blood loss, excised spleen weight, complications, and hospital stays. RESULTS: A total of 216 splenectomies (135 OS and 81 LS) were performed from April 1999 to March 2007. Five laparoscopic cases were converted to open surgery owing to operative bleeding or bleeding of splenic fossa. The other 76 patients were performed LSs successfully. No major operative complications occurred. There was no operative death. Excised spleen weight >400 g was present in 56% of cases in this series. At 7 days postoperatively, the platelet counts, white blood cell counts, and hemoglobin significantly increased after open and laparoscopic surgeries, and increase of alanine aminotransferase, aspartate aminotransferase, total bilirubin, and directed bilirubin of LS were significantly different with open cases. Operation times of LS and OS were 2.9+/-0.7 hours and 2.6+/-0.6 hours, respectively. Blood losses were 150.6+/-135.4 mL and 633.8+/-340.3 mL (P<0.01), excised spleen weights were 585.7+/-184.6 g and 591.1+/-153.4 g (P>0.05), and hospital stay were 8.2+/-2.0 days and 11.9+/-3.8 days (P<0.01). Operative associated complications were noted in both LS and OS. Less blood loss, shorter hospital stay, and less impairment of liver function were observed in LS than OS. CONCLUSIONS: LS is feasible, effective, and safe procedures for hypersplenism secondary to liver cirrhosis and contributes to less impairment of liver function, less blood loss, and shorter hospital stay.

CTLA4Ig introduced by adenovirus vector locally to prolong the survival of xenogeneic skin grafts on rat burn wounds.

BACKGROUND: The purpose of this study was to explore an applicable approach for prolonging the survival of heterogenetic skin grafts on burn wounds with CTLA4Ig. METHODS: An adenovirus vector named Ad-CTLA4Ig, which could express human CTLA4Ig fusion protein, was constructed. Infecting and replicating in 293 cells, more Ad-CTLA4Ig and recombinant human CTLA4Ig (rhCTLA4Ig) were prepared, respectively. In a rat flame thermal injury model, the effect of rhCTLA4Ig on survival time of human skin graft on the eschar-excised rat burn wound was observed. Meanwhile, the efficiency of Ad-CTLA4Ig infecting cultured skin fibroblasts, keratinocytes, and partial-thickness skin samples were checked by CTLA4Ig expression essay. Then, the Ad-CTLA4Ig was administered locally on the eschar-excised wound and dermis of the skin graft, and the survival time of the human skin graft on burn wound was measured. The influence of the systemic immune function by rhCTLA4Ig and Ad-CTLA4Ig were also determined. RESULTS: The prepared rhCTLA4Ig from the supernatant of Ad-CTLA4Ig-infected 293 cells was verified by sodium dodecyl sulfate polyacrylamide gel electrophoresis sodium dodecyl sulfate poly-acrylamide gel electrophoresis and Western blot. It was found that CTLA4Ig could significantly prolong the xenogeneic skin graft survival in a dosage-dependent manner. Interestingly, the survival time was longer when CTLA4Ig was used 24 hours posttransplantation than that at hour 0. The expression of CTLA4Ig could be observed in the cultured skin fibroblasts, keratinocytes, and skin pieces soon after Ad-CTLA4Ig transfection, as demonstrated by either immunocellular chemistry or immunohistochemistry assay. When Ad-CTLA4Ig was locally administered during skin transplantation on burn wound, the survival time was increased from 7.9 days of control group to 21.6 days, whereas the systemic immune function was not affected. CONCLUSION: Administration of Ad-CTLA4Ig locally could prolong the survival time of xenogeneic skin graft on burn wound without significantly influencing the systemic immune function.

[The influence of the application of cytotoxic lymphocyte antigen 4-Ig adenovirus on the burn wounds with alloskin grafting on the murine immune function].

OBJECTIVE: To investigate the influence of local application of cytotoxic lymphocyte antigen 4-Ig (CTLA4-Ig) adenovirus on the burn wound with alloskin grafting upon the murine immune function. METHODS: Sixty BALB/c mice were randomly divided into A (operation control), B (CTLA4-Ig transfection) and C (normal control) groups, with 20 mice in each group. Skin wounds (full-thickness loss) sized 1.5 cm x 1.5 cm were created on the backs of mice in A and B groups. Then the skin grafts of the same size obtained from C57BL mice were grafted into the skin wounds. 0.1 g of cross-linking polyacrylic resin (carbomer cream) without adenovirus was daubed onto the wounds in A group, and the same amount of carbomer cream with adenovirus in titers of 5 x 10(9)/L was daubed onto the wounds in B group, while no treatment was given in C group. 1 ml of 10% SRBC (sheep red blood cell) was injected intraperitoneally to all the mice of the three groups on the 1st post injury day (PID). Splenocytes from BALB/c, C57BL and Kunming mice were harvested for mixed lymphocyte culture on 7, 14, 21 and 28 PIDs. Agglutination assay was used in the same time to detect the SRBC antibody titers. RESULTS: The reaction of murine splenocytes in B group to the donor (C57BL) splenocytes was suppressed in a specific way (P < 0.05) within 14 PIDs. There was no difference in the titers of anti-SRBC antibody among the 3 groups (P > 0.05). CONCLUSION: Local application of CTLA4-Ig recombinant adenovirus exhibited no influence on the murine humoral immunity, but might induce systemic and specific T cell tolerance in immunity system.