Elevated Transient Receptor Potential Melastatin 8 (TRPM8) Expression Is Correlated with Poor Prognosis in Pancreatic Cancer.

BACKGROUND The transient receptor potential melastatin 8 (TRPM8) was found to be expressed abnormally in a variety of tumors and is associated with unfavorable prognosis in human cancers. However, its clinical significance in pancreatic cancer (PC) is mostly unknown. MATERIAL AND METHODS qRT-PCR was performed to measure the expression of TRPM8 in 110 pairs of PC tissues and the adjacent non-cancerous tissues. The association of TRPM8 expression with the clinical characters of PC patients was analyzed using the chi-square test. Furthermore, the prognostic value of TRPM8 was determined with Kaplan-Meier survival curve and Cox regression analysis. RESULTS We found that the expression level of TRPM8 was significantly elevated in PC tissues compared to the non-cancerous controls (P<0.001). In addition, a close relationship was observed between elevated TRPM8 expression with large tumor size (P=0.001), advanced TNM (P=0.013), and distant metastasis (P=0.034). Survival analysis suggested that patients with high TRPM8 expression has worse OS (P=0.001) and DFS (P<0.001) than those with low TRPM8 expression. Moreover, TRPM8 was confirmed as a valuable prognostic biomarker for OS (HR=1.913; 95% CI: 1.020-3.589; P=0.043) or DFS (HR=2.374; 95% CI: 1.269-4.443; P=0.007) of PC patients. CONCLUSIONS This study shows that TRPM8 expression is significantly up-regulated in PC and it might be a useful prognostic factor for patients with PC.

MicroRNA­506 participates in pancreatic cancer pathogenesis by targeting PIM3.

MicroRNA (miRNA) is a type of short non-coding RNA that suppresses the expression of protein coding genes by partial complementary binding to the 3'­untranslated regions (UTRs) of mRNAs. miRNA expression alterations are involved in the initiation, progression and metastasis of human cancer and it has been suggested that miRNAs function as tumor suppressors as well as oncogenes in cancer development. PIM-3 is a member of the proto-oncogene PIM family, the aberrant expression of which exists in human pancreatic cancer tissues. There are reports indicating that overexpression of PIM3 is associated with the promotion of pancreatic cancer cell proliferation. The aim of the present study was to identify micro (mi)RNAs that regulate the expression of the oncogene PIM3 in PC. It was confirmed that the expression of PIM3 was regulated by miRNAs through an AGO2 knockout experiment. Subsequently, a dual luciferase assay system was constructed and used to screen 13 selected miRNAs that may target the PIM3 3'UTR directly. The results indicated that miR­15a/b, miR­16, miR­33a/b, miR­124, miR­195 and miR­506 repressed the luciferase activity by targeting the PIM3 3'UTR. However, only the expression of miR­506 was negatively correlated with PIM3 expression in PC tissues (r=­0.38, P=0.017). Furthermore, a biological functional study indicated that miR­506 functioned as a tumor suppressor by repressing PC cell proliferation, which was partially reversed by PIM3 overexpression. To the best of our knowledge, the present study was the first to reveal the tumor suppressor function of miR­506 in PC, which has the potential to be employed in the diagnosis and treatment of PC.

Effects of intensive insulin therapy combined with low molecular weight heparin anticoagulant therapy on severe pancreatitis.

The current study explored the effects of intensive insulin therapy (IIT) combined with low molecular weight heparin (LMWH) anticoagulant therapy on severe acute pancreatitis (SAP). A total of 134 patients with SAP that received treatment between June 2008 and June 2012 were divided randomly into groups A (control; n=33), B (IIT; n=33), C (LMWH; n=34) and D (IIT + LMWH; n=34). Group A were treated routinely. Group B received continuous pumped insulin, as well as the routine treatment, to maintain the blood sugar level between 4.4 and 6.1 mmol/l. Group C received a subcutaneous injection of LMWH every 12 h in addition to the routine treatment. Group D received IIT + LMWH and the routine treatment. The white blood cell count, hemodiastase, serum albumin, arterial partial pressure of oxygen and prothrombin time were recorded prior to treatment and 1, 3, 5, 7 and 14 days after the initiation of treatment. The intestinal function recovery time, incidence rate of multiple organ failure (MOF), length of hospitalization and fatality rates were observed. IIT + LMWH noticeably increased the white blood cell count, hemodiastase level, serum albumin level and the arterial partial pressure of oxygen in the patients with SAP (P<0.05). It markedly shortened the intestinal recovery time and the length of stay and reduced the incidence rate of MOF, the surgery rate and the fatality rate (P<0.05). It did not aggravate the hemorrhagic tendency of SAP (P>0.05). IIT + LMWH had a noticeably improved clinical curative effect on SAP compared with that of the other treatments.

Modified hepatic vascular occlusion facilitates hepatectomy for malignant hepatic tumor in laparoscopy.

PURPOSE: The parenchymal transection and bleeding are important problems in laparoscopic hepatectomy (LH). The study aimed to evaluate the feasibility and safety of LH for a malignant hepatic tumor (MHT) with hepatic vascular occlusion (HVO) only using a monopolar electrocautery. METHODS: A total of 31 patients' profiles, operative data, clinical outcomes, pathologic findings, and follow-up information were collected who underwent LH with HVO. RESULTS: The median operative time was 176 minutes and the median postoperative hospital stay was 9 days. The median estimated blood loss was 310 mL. The median tumor size was 4.6 cm and the median surgical margin was 12 mm.The ischemia injury to the liver in patients subjected to selective HVO was less than that in patients subjected to the Pringle maneuver. CONCLUSIONS: HVO facilitates LH in transecting liver parenchyma and reduces bleeding, making LH safe and feasible only using monopolar electrocautery in selected patients with malignancies. Moreover, the selective HVO have more advantages over the Pringle maneuver in decreasing ischemic injury.

Retroperitoneal laparoscopic debridement and drainage of infected retroperitoneal necrosis in severe acute pancreatitis.

OBJECTIVE: To explore the effect of retroperitoneal laparoscopic debridement and drainage on infected necrosis in severe acute pancreatitis. MATERIALS AND METHODS: This retrospective study included 18 patients with severe acute pancreatitis (SAP) undergoing retroperitoneal laparoscopic debridement and drainage from May 2006 to April 2012 in our hospital. All patients had infected retroperitoneal necrosis and single or multiple peritoneal abscesses. Eleven patients transferred to our hospital were treated with the retroperitoneal laparoscopic debridement and drainage within 24-72 hours after admission. Conservative treatments were given to eight patients. Retroperitoneal laparoscopic debridement and drainage were applied 3-11 days after admission. RESULTS: All patients had infection of necrotic pancreas or peripancreatic tissues. Twelve patients had organ failure. Three patients underwent secondary surgery. Laparotomy with debridement and drainage were applied to one patient who had a huge lesser sac abscess 7 days after first surgery. The other two patients were given secondary retroperitoneal laparoscopic debridement and drainage. One case was complicated by retroperitoneal hemorrhage, four cases had pancreatic leakage, and no intestinal fistula was found. The patients' heart rate, respiration, temperature, and white blood cell count were significantly improved 48 hours after surgery compared with those prior to surgery (p<0.05). The average length of stay in hospitals was 40.8 days (range, 6-121 days), and the drainage tube indwelling time was 44.4 days (range, 2-182 days). CONCLUSION: Retroperitoneal laparoscopic debridement and drainage is an SAP surgical treatment with a minimally invasive procedure and a good effect, and can be applied for infected retroperitoneal necrosis in early SAP.

Laparotomy versus retroperitoneal laparoscopy in debridement and drainage of retroperitoneal infected necrosis in severe acute pancreatitis.

BACKGROUND: The aim of this study was to compare laparotomy and retroperitoneal laparoscopy in debridement and drainage of retroperitoneal infected necrosis of severe acute pancreatitis (SAP), and to evaluate the curative efficacy and the timing of retroperitoneal laparoscopic debridement drainage (RLDD) for SAP patients. METHODS: We performed a retrospective analysis of 50 SAP cases, including 18 patients in the RLDD group and 32 patients in the laparotomy group. Observed indices included gender, age, CT severity index, Ranson score, APACHE II score, preoperative course, length of stay, operation time, mortality, postoperative complications, drainage tube indwelling time, and change of body temperature and peripheral white blood cell (PWBC) count between the time before the operation and at 48 h after surgery. RESULTS: Between the RLDD group and the laparotomy group, there was a significant difference in operation time (130 ± 15 vs. 148 ± 25 h; P = 0.007), length of stay [40.8 (6-121) vs. 55.9 (28-133) days; P = 0.053], and preoperative course [14.7 (5-31) vs. 18.3 (6-31) days; P = 0.05], but no significant difference in average drainage tube indwelling time [44.4 (2-182) vs. 49.8 (2-175) days; P = 0.663]. More improvement in body temperature and PWBC count was observed in the patients of the RLDD group. There was one death (1/18) in the RLDD group and four (4/32) in the laparotomy group. Fourteen cases (14/32) in the laparotomy group had postoperative complications, including pancreatic fistula (n = 11), intestinal fistula (n = 2), retroperitoneal hemorrhage (n = 2), infection of incision (n = 9), and 5 cases (5/18) in the RLDD group, including pancreatic fistula (n = 4) and retroperitoneal hemorrhage (n = 1). CONCLUSIONS: RLDD, as minimally invasive surgery, is technically feasible, safe, and effective in the treatment of retroperitoneal infected necrosis in SAP patients, in contrast to the laparotomy technique, and can be performed in the early phase of SAP to prevent the deterioration of the disease.

Hepatitis B virus X protein represses miRNA-148a to enhance tumorigenesis.

MicroRNAs (miRNAs) have been shown to be dysregulated in virus-related cancers; however, miRNA regulation of virus-related cancer development and progression remains poorly understood. Here, we report that miR-148a is repressed by hepatitis B virus (HBV) X protein (HBx) to promote cancer growth and metastasis in a mouse model of hepatocellular carcinoma (HCC). Hematopoietic pre-B cell leukemia transcription factor-interacting protein (HPIP) is an important regulator of cancer cell growth. We used miRNA target prediction programs to identify miR-148a as a regulator of HPIP. Expression of miR-148a in hepatoma cells reduced HPIP expression, leading to repression of AKT and ERK and subsequent inhibition of mTOR through the AKT/ERK/FOXO4/ATF5 pathway. HBx has been shown to play a critical role in the molecular pathogenesis of HBV-related HCC. We found that HBx suppressed p53-mediated activation of miR-148a. Moreover, expression of miR-148a was downregulated in patients with HBV-related liver cancer and negatively correlated with HPIP, which was upregulated in patients with liver cancer. In cultured cells and a mouse xenograft model, miR-148a reduced the growth, epithelial-to-mesenchymal transition, invasion, and metastasis of HBx-expressing hepatocarcinoma cells through inhibition of HPIP-mediated mTOR signaling. Thus, miR-148a activation or HPIP inhibition may be a useful strategy for cancer treatment.

FHL family members suppress vascular endothelial growth factor expression through blockade of dimerization of HIF1α and HIF1ß.

Four and a half LIM domain (FHL) proteins belong to a family of LIM-only proteins that have been implicated in the development and progression of various types of cancers. However, the role of FHL proteins in tumor angiogenesis remains to be elucidated. Herein, we demonstrate that FHL1-3 decrease the promoter activity and expression of vascular endothelial growth factor (VEGF), the key regulator of angiogenesis in cancer growth and progression as well as an important target gene of the transcription factor hypoxia-inducible factor 1 (HIF1α/HIF1ß). FHL1-3 interacted with HIF1α both in vitro and in vivo. A single LIM domain of FHL1 was sufficient for its interaction with HIF1α. FHL1 interacted with the HIF1α region containing basic helix-loop-helix (bHLH) motif and PER-ARNT-SIM domain, both of which aid in dimerization with HIF1ß and DNA binding. FHL1-3 inhibited HIF1 transcriptional activity and HIF1-mediated VEGF expression in a hypoxia-independent manner. Moreover, FHL1 blocked HIF1α-HIF1ß heterodimerization and HIF1α recruitment to the VEGF promoter. These data suggest that FHL proteins are involved in negative regulation of VEGF possibly by interfering with the dimerization and DNA binding of HIF1 subunits and may play an important role in tumor angiogenesis.

[Developmental History of laparoscopic liver resection].

Since 1991, the first laparoscopic liver resection (LLR) was reported by Reich, LLR had been applied and improved by different scholars like Wayand (1993), Ferzli (1995), Cherqui (2002), Descottes (2003). Zhou Weiping reported the first LLR in China in 1994. In 2004, after ten years development, LLR became more and more mature and international exchange happens. In 2006, indication and contraindications of LLR were formulated internationally, which promote the application and spread of LLR, especially after 2007. The advantages of LLR include minimal invasion, faster recovery and less complications, but shortages still exist, and the improvements relies on development of related techniques in terms of Laparoscopic ultrasound, LUS, liver dissecting sealer, electrocoagulation electrotome, parenchymatous organ dissection and in vitro simulator. Therefore LLR can get new progress.

[The clinical study of percutaneous transhepatic radiofrequency ablation combined with tumor edge of percutaneous absolute ethanol injection on liver cancer adjacent to major blood vessels].

OBJECTIVE: To explore the effects of percutaneous transhepatic radiofrequency ablation (PRFA) combined with tumor edge of percutaneous absolute ethanol injection (PEI) on liver cancer adjacent to major blood vessels. METHODS: Seventy five patients with liver cancer adjacent to major blood vessels were randomly divided into two groups: PRFA+PEI therapy group (38 cases) and PRFA control group (37 cases). Tumor necrosis rate, AFP levels, local recurrence rate, median for survival time and cum survival were used as the evaluation index to evaluate the efficacies of the two methods. RESULTS: Tumor necrosis rates of the therapy group and the control group were 84.2% and 54.1% (P < 0.01), respectively; AFP levels of therapy group and control group at 1, 3, 6 and 12 months after treatment were (105.0 ± 35.5) µg/L, (28.4 ± 4.3) µg/L, (58.6 ± 6.7) µg/L, (89.5 ± 12.5) µg/L and (137.2 ± 34.6) µg/L, (84.2 ± 18.4) µg/L, (106.6 ± 20.3) µg/L, (173.7 ± 32.0) µg/L, respectively. The rates of therapy group was significantly lower than of control group. Local recurrence rates of the therapy group and control group were 2.6%, 7.9%, 13.2% and 31.6% vs 10.8%, 21.6% , 40.5% and 62.1% (P < 0.05) at 3, 6, 12 and 24 months after treatment, respectively. Median for survival time of the therapy group and control group were 28.0 ± 2.8 months and 19.0 ± 3.6 months, respectively. Cum survival of the therapy group and control group were 84.2%, 78.9%, 60.5% and 31.6% vs 78.4%, 67.6%, 37.8% and 8.1% (P < 0.05) at 6, 12, 24 and 36 months after treatment, respectively. CONCLUSION: PEI as a supplementary treatment of PRFA can effectively improve the treatment of liver cancer adjacent to major blood vessels and significantly reduce the local recurrence rate and improve long-term survival rates.

Laparoendoscopic single-site liver resection: a preliminary report of 12 cases.

BACKGROUND: Laparoendoscopic single-site (LESS) surgery is an emerging laparoscopic procedure previously used for cholecystectomy and appendectomy. However, few studies have examined LESS liver resection, and its benefits require investigation. This study aimed to evaluate the feasibility and safety of LESS liver resection. METHODS: From December 2009 to October 2010, 12 patients were selected for LESS liver resection with institutional review board approval. The LESS technique was performed using a transumbilical TriPort or three 5-mm trocars with a 5-mm linear or flexible laparoscope. Conventional or articulating laparoscopic instruments were used to mobilize and transect the lesions. RESULTS: The LESS liver resection procedure was successfully completed for 10 patients (83.3%), with the remaining 2 patients (16.7%) undergoing conversion to conventional multiport laparoscopy. The procedures consisted of left lateral segment resection (n = 4) and partial resection (n = 8) in addition to concomitant cholecystectomy (n = 3). The mean operative time was 80.4 min (range, 35-160 min), and the mean estimated blood loss was 45 ml (range, 20-800 min). No postoperative complications were noted except for biliary leakage (200 ml/day)in one patient. The mean hospital stay was 4.3 days (range, 2-8 days). No patient required postoperative analgesia, and the pain visual analog score 48 h after surgery was 0.53 (range, 0-2). Pathology identified 10 benign and 2 malignant liver tumors with a clear margin. CONCLUSIONS: Our preliminary data show that LESS liver resection is safe and feasible for selected patients, with potential benefits that include a fast recovery, light pain, and cosmetically acceptable scarring. However, this procedure requires advanced instruments and complicated laparoscopic techniques, with a risk of intraoperative bleeding and postoperative bile leakage.

Clinical effects of intensive insulin therapy treating traumatic shock combined with multiple organ dysfunction syndrome.

The therapeutic effects of intensive insulin therapy in treatment of traumatic shock combined with multiple organ dysfunction syndrome (MODS) were investigated. A total of 114 patients with traumatic shock combined with MODS were randomly divided into two groups: control group (n=56) treated with conventional therapy, and intensive insulin therapy group (n=58) treated with conventional therapy plus continuous insulin pumping to control the blood glucose level at range of 4.4-6.1 mmol/L. White blood cells (WBC) counts, prothrombin time (PT), serum creatinine (SCr), alanine aminotransferase (ALT), serum albumin and PaO(2) were measured before and at the day 1, 3, 5, 7 and 14 after treatment. The incidence of gastrointestinal dysfunction, the incidence of MODS, hospital stay and the mortality were also observed and compared. After intensive insulin therapy, the WBC counts, SCr, ALT and PT were significantly reduced (P<0.05), but the level of serum albumin was significantly increased (P<0.05) at the day 3, 5, 7 and 14. In the meantime, the PaO2 was significantly elevated at the day 3, 5 and 7 (P<0.01) after intensive insulin therapy. The incidence of gastrointestinal dysfunction, the incidence of MODS, the length of hospital stay and the mortality were markedly decreased (P<0.01). The results suggest early treatment with intensive insulin therapy is effective for traumatic shock combined with MODS and can decrease the length of hospital stay and the mortality.

Clinical Effects of Intensive Insulin Therapy Treating Traumatic Shock Combined with Multiple Organ Dysfunction Syndrome / 华中科技大学学报（医学）（英德文版）

The therapeutic effects of intensive insulin therapy in treatment of traumatic shock combined with multiple organ dysfunction syndrome (MODS) were investigated.A total of 114 patients with traumatic shock combined with MODS were randomly divided into two groups:control group (n=56) treated with conventional therapy,and intensive insulin therapy group (n=58) treated with conventional therapy plus continuous insulin pumping to control the blood glucose level at range of 4.4-6.1 mmol/L.White blood cells (WBC) counts,prothrombin time (PT),serum creatinine (SCr),alanine aminotransferase (ALT),serum albumin and PaO2 were measured before and at the day 1,3,5,7 and 14 after treatment.The incidence of gastrointestinal dysfunction,the incidence of MODS,hospital stay and the mortality were also observed and compared.After intensive insulin therapy,the WBC counts,SCr,ALT and PT were significantly reduced (P＜0.05),but the level of serum albumin was significantly increased (P＜0.05) at the day 3,5,7 and 14.In the meantime,the PaO2 was significantly elevated at the day 3,5 and 7 (P＜0.01) after intensive insulin therapy.The incidence of gastrointestinal dysfunction,the incidence of MODS,the length of hospital stay and the mortality were markedly decreased (P＜0.01).The results suggest early treatment with intensive insulin therapy is effective for traumatic shock combined with MODS and can decrease the length of hospital stay and the mortality.

[Effects of anticoagulation therapy with low molecular weight heparin in acute pancreatitis].

OBJECTIVE: To evaluate the effects of anticoagulation therapy with low molecular weight heparin in acute pancreatitis. METHODS: Low molecular weight heparin, in a dose of 40 mg or 0.01 ml/kg, by subcutaneous injection, every 12 hours, was administered to 17 acute pancreatitis patients combined with conventional therapy. The changes of serum enzymology and prognosis in patients treated with low molecular weight heparin or conventional therapy were observed. RESULTS: Anticoagulation by low molecular weight heparin could significantly decrease the blood white cell count of patients with acute pancreatitis and increase their arterial blood oxygen partial pressure. It could cut down the duration of hospitalization and reduce the aggravation rate, secondary operation rate, and mortality of these patients without increasing hemorrhagic tendency or its related complications. CONCLUSION: Anticoagulation therapy with low molecular weight heparin is safe and effective in the treatment of acute pancreatitis, and it may improve its prognosis.

[Expression of a novel human interleukin-13 in E.coli and its biological activity assay].

AIM: To express a novel Gln98-deleted human interleukin-13 in E.coli. METHODS: Total RNA was isolated from Jurkat cells costimulated with PHA and ConA. A 358 bp-specific DNA fragment encoding hIL-13 was amplified by semi-nested RT-PCR. DNA sequencing showed that the target DNA was a Gln98-deleted novel splicing of hIL-13. This hIL-13 cDNA and plasmid pBV220 were ligated at BamH I and EcoR I sites to construct the expression vector. After transforming E.coli strain DH5alpha, the expression of the novel splicing hIL-13 gene was induced by shifting culture temperature from 30 degrees Celsius to 42 degrees Celsius. The expression product was then purified by chromatography on Sepharcryl-200 gel column, and the bioactivity was detected by MTT colorimetry on the growth of TF-1 cell line. RESULTS: The novel rhIL-13 was expressed in the form of inclusion bodies. After purification and renaturation, the specific activity of the novel rhIL-13 was 1.6x10(6) IU/mg. CONCLUSION: The novel rhIL-13 with biological activity has been obtained, which lays the foundation for treating cancer and septicemia by the cytokine in future.