Forkhead box M1 (FOXM1) transcription factor is a key oncogenic driver of aggressive human meningioma progression.

AIMS: Aggressive meningioma remains incurable with neither chemo- nor targeted therapies proven effective, largely due to unidentified genetic alterations and/or aberrant oncogenic pathways driving the disease progression. In this study, we examined the expression and function of Forkhead box M1 (FOXM1) transcription factor during meningioma progression. METHODS: Human meningioma samples (n = 101) were collected, followed by Western blotting, quantitative PCR, immunohistochemical and progression-free survival (PFS) analyses. For in vitro assays, FOXM1 was overexpressed or knocked-down in benign (SF4433 and SF4068) or malignant (SF3061 and IOMM-Lee) human meningioma cell lines respectively. For in vivo studies, siomycin A (a FOXM1 inhibitor)-pretreated or control IOMM-Lee cells were implanted subcutaneously in nude mice. RESULTS: FOXM1 expression was increased in higher grades of meningioma and correlated with the mitotic index in the tumour tissue. Moreover, FOXM1 was increased in recurrent meningioma compared with the matched primary lesions. The patients who had higher FOXM1 expression had shorter PFS. In the subsequent in vitro assays, knockdown of FOXM1 in malignant meningioma cell lines resulted in decreased tumour cell proliferation, angiogenesis and invasion, potentially via regulation of ß-catenin, cyclin D1, p21, interleukin-8, vascular endothelial growth factor-A, PLAU, and epithelial-to-mesenchymal transition-related genes, whereas overexpression of FOXM1 in benign meningioma cell lines had the opposite effects. Last, suppression of FOXM1 using a pharmacological inhibitor, siomycin A, decreased tumour growth in an in vivo mouse model. CONCLUSIONS: Our data demonstrate that FOXM1 is a key transcription factor regulating oncogenic signalling pathways in meningioma progression, and a promising therapeutic target for aggressive meningioma.

Effects of a finger exercise program on hand function in automobile workers with hand osteoarthritis: A randomized controlled trial.

Hand osteoarthritis reduces a person's ability to perform work activities and return to their occupation. We investigated the effects of a finger exercise program on hand grip strength, pain, physical function, and stiffness in automobile manufacturing workers with hand osteoarthritis. This randomized controlled trial was conducted on 29 subjects. Fifteen experimental subjects received a finger exercise program with paraffin baths, while 14 control subjects received only paraffin baths. Hand grip strength, pain, physical function, and stiffness were assessed at baseline and 8 weeks later. In the experimental group, hand grip strength (P < 0.001) and Australian/Canadian osteoarthritis hand index (AUSCAN) scores (pain, P < 0.001; stiffness, P < 0.001; physical function, P < 0.001) were significantly improved by 3.52 ± 2.03, 21.6 ± 8.3 (pain), 16.8 ± 10.21 (stiffness), and 13.86 ± 4.54 (physical function) compared with preintervention values. In the control group, hand grip strength (P = 0.004) and AUSCAN scores (pain, P < 0.001; stiffness, P = 0.019; physical function, P < 0.001) were significantly improved by 0.57 ± 0.62, 7.85 ± 5.46 (pain) 11.42 ± 7.18 (stiffness), and 10.28 ± 14.41 (physical function) compared with preintervention values. Significant differences between groups were found for postintervention hand grip strength (P = 0.015) and AUSCAN index subscale scores (pain, P < 0.001; physical function, P = 0.020). A combined finger exercise and paraffin bath program is effective in reducing pain, improving physical function, and increasing hand grip strength in workers with hand osteoarthritis.

Transcatheter arterial embolization for iatrogenic bleeding after endoscopic ultrasound-guided pancreaticobiliary drainage.

PURPOSE: The purpose of this study was to report the incidence of massive bleeding after endoscopic ultrasound-guided transmural pancreaticobiliary drainage (EUS-TPBD) and the clinical outcomes in patients with this condition treated with transcatheter arterial embolization (TAE). PATIENTS AND METHODS: We performed a 9-year retrospective analysis of 797 EUS-TPBD procedures (excluding gallbladder or pseudocysts) in 729 patients. Among them, twelve (12/729, 1.65%) patients were referred for TAE to manage active bleeding adjacent to the TPBD sites. There were 8 men and 4 women with a mean age of 66.1 years±13.4 (SD) (range: 45-89 years). The clinical and procedure data of these 12 patients were reviewed. RESULTS: Thirteen TAE procedures in 12 patients were performed. The bleeding sites were the left hepatic artery (n=7), the right hepatic artery (n=3), the left gastric artery (n=1), the left accessory gastric artery (n=1) and gastroduodenal artery (n=1). TAE was performed with gelatin sponge particles (n=1), coil (n=1) and n-butyl-2 cyanoacrylate with/without coils (n=11), with technical and clinical success rates of 100% (13/13) and 85% (11/13), respectively. Re-bleeding following embolization with gelatin sponge particles occurred in one patient. Procedure-related ischemic hepatitis was observed in another patient with pancreatic cancer with portal vein involvement. CONCLUSION: On the basis of our results, TAE using n-butyl-2 cyanoacrylate seems safe and effective for the treatment of bleeding after EUS-TPBD procedures. When the portal vein is compromised, TAE of the hepatic artery can cause ischemic liver damage.

Effects of scalp nerve block on pain and emergence agitation after paediatric nevus surgery: a clinical trial.

BACKGROUND: Pain is considered as being one cause of post-operative emergence agitation (EA) from sevoflurane anaesthesia. The purpose of this study was to investigate the pure effect of post-operative pain on EA after sevoflurane anaesthesia in preschool children undergoing excision of scalp nevi. METHODS: Forty-four children, 1-7 years old, undergoing scalp nevus excision were enrolled. Patients were randomly assigned to two groups: the remifentanil group received single intravenous injection of short-acting synthetic opioid, remifentanil 1 µg/kg just before the scalp incision, and the block group received scalp nerve block with 0.25% ropivacaine after intubation. The end-tidal sevoflurane concentration was maintained around 1.5 vol% unless the mean arterial pressure is out of ±20% range of preoperative values during surgery in both groups. Watcha behaviour scale for EA and face, legs, activity, cry, consolability (FLACC) scale scores for pain were recorded post-operatively. RESULTS: There was no difference in end-tidal sevoflurane concentration between the two groups during surgery and the emergence period. Agitation incidence and scores were not different between the two groups during the recovery period. FLACC scale was significantly lower in the block group than in the remifentanil group at post-anaesthesia care unit (PACU) arrival, at 10 and 20 min after PACU arrival, respectively. CONCLUSION: The scalp nerve block decreased the early post-operative pain after paediatric nevus excision, but it did not decrease the incidence of EA with sevoflurane anaesthesia.

GABA (γ-Aminobutyric Acid) Uptake Via the GABA Permease GabP Represses Virulence Gene Expression in Pseudomonas syringae pv. tomato DC3000.

The nonprotein amino acid γ-aminobutyric acid (GABA) is the most abundant amino acid in the tomato (Solanum lycopersicum) leaf apoplast and is synthesized by Arabidopsis thaliana in response to infection by the bacterial pathogen Pseudomonas syringae pv. tomato DC3000 (hereafter called DC3000). High levels of exogenous GABA have previously been shown to repress the expression of the type III secretion system (T3SS) in DC3000, resulting in reduced elicitation of the hypersensitive response (HR) in the nonhost plant tobacco (Nicotiana tabacum). This study demonstrates that the GABA permease GabP provides the primary mechanism for GABA uptake by DC3000 and that the gabP deletion mutant ΔgabP is insensitive to GABA-mediated repression of T3SS expression. ΔgabP displayed an enhanced ability to elicit the HR in young tobacco leaves and in tobacco plants engineered to produce increased levels of GABA, which supports the hypothesis that GABA uptake via GabP acts to regulate T3SS expression in planta. The observation that P. syringae can be rendered insensitive to GABA through loss of gabP but that gabP is retained by this bacterium suggests that GabP is important for DC3000 in a natural setting, either for nutrition or as a mechanism for regulating gene expression. [Formula: see text] Copyright © 2016 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license .

Evaluation of in vivo antitumor effects of ANT2 shRNA delivered using PEI and ultrasound with microbubbles.

Gene therapy using RNA interference can be directed against tumors through various strategies, but has been hindered owing to the inefficiency of non-viral delivery. To evaluate the antitumor effects of adenine nucleotide translocase-2 (ANT2) short hairpin RNA (shRNA) by intraperitoneal injection using the polyethylenimine (PEI) and an ultrasound gene delivery method, human breast carcinoma MDA-MB-231 cells were injected subcutaneously into NOG (NOD/Shi-scid/IL-2Rγ(null)) mice. The results showed greater tumor regression (*P<0.05) as well as an increased survival rate in the group receiving ANT2 shRNA+two types of enhancer relative to the groups receiving ANT2 shRNA without enhancer. These findings demonstrate that the introduction of PEI and ultrasound with SonoVue exerted enhanced antitumor effects in vivo. Although the combination of jet-PEI and ultrasound provided the best results with respect to tumor regression, the antitumor effects from the individual enhancers were approximately equivalent. In addition, we confirmed that there was no toxicity on aspartate aminotransferase and alanine aminotransferase levels in the liver and albumin, blood urea nitrogen or creatine kinase levels in the kidney following the various gene delivery methods.

Primary and revision efficacy of cross-wired metallic stents for endoscopic bilateral stent-in-stent placement in malignant hilar biliary strictures.

BACKGROUND AND STUDY AIMS: Endoscopic bilateral drainage for inoperable malignant hilar biliary strictures (HBS) using metal stents is considered to be technically difficult. Furthermore, endoscopic revision of bilateral stenting after occlusion can be challenging. This study was performed to evaluate the long-term efficacy of endoscopic bilateral stent-in-stent placement of cross-wired metallic stents in high-grade malignant HBS and planned endoscopic bilateral revision. PATIENTS AND METHODS: A total of 84 patients with inoperable high-grade malignant HBS were enrolled from three academic tertiary referral centers. Two cross-wired metal stents were inserted using a bilateral stent-in-stent placement method. Bilateral endoscopic revision was also performed during follow-up using either identical metal stents or plastic stents. The main outcome measurements were technical and functional success, complications, stent patency, and endoscopic revision efficacy. RESULTS: The technical and clinical success rates of endoscopic bilateral stent-in-stent placement of cross-wired metallic stents were 95.2% (80/84) and 92.9% (78/84), respectively. Median patency (range) and survival were 238 days (10-429) and 256 days (10-1130), respectively. Obstruction of primary bilateral stents occurred in 30.8% (24/78) of patients with functionally successful stent placement. The technical and clinical success rates of planned bilateral endoscopic revision for occluded stents were 83.3% (20/24) and 79.2% (19/24), respectively. For revision, bilateral metallic stents were placed in 11 patients (55.0%); the remaining patients received plastic stents. CONCLUSIONS: Palliative endoscopic bilateral stent-in-stent placement of cross-wired metallic stents was effective in patients with inoperable HBS. Revision endoscopic bilateral stenting may be feasible and successful in cases where the primary deployed metal stents are occluded.

Endoscopic ultrasound-guided rendezvous for biliary access after failed cannulation.

INTRODUCTION: Selective cannulation fails in approximately 3 % of endoscopic retrograde cholangiography (ERC) procedures. An endoscopic ultrasound-guided rendezvous technique (EUS - RV) may salvage failed cannulation. The aims of the current study were to determine the safety and efficacy of EUS - RV. METHODS: A total of 40 patients underwent salvage EUS - RV. EUS - RV was attempted immediately after failed biliary cannulation. A dilated intra- or extra-hepatic biliary duct (IHBD or EHBD) was punctured from the stomach or the small intestine under EUS guidance followed by cholangiography and antegrade manipulation of the guide wire into the small intestine. Finally, the echoendoscope was exchanged for an appropriate endoscope and biliary cannulation was achieved over or adjacent to the guide wire. RESULT: EUS-RV appears safe and effective and may be considered as a primary salvage technique after failed cannulation. Antegrade manipulation of the guide wire into the small intestine was achieved in 29 of 40 patients (73 %; EHBD 25 /31 and IHBD 4/9). The reasons for failure were inability to advance the guide wire through an obstruction or a native ampulla. Re-attempt at ERC immediately after failed EUS - RV was made in seven of the 11 patients, and was successful in four. The remaining seven patients underwent percutaneous drainage within 3 days. Complications occurred in five patients (13 %), including pancreatitis, abdominal pain, pneumoperitoneum, and sepsis/death, which was unlikely to be related to the procedure. CONCLUSION: EUS - RV is safe and effective and should be considered as a primary salvage technique after failed cannulation. Immediate re-attempt at ERC after failed EUS - RV is warranted, as EUS-guided cholangiogram can facilitate biliary cannulation in some cases. Finally, prompt alternative biliary drainage should be available.

Conjunctival pigmentation after 23-gauge microincisional vitrectomy surgery.

AIM: To investigate cases with conjunctival pigmentation originating from sclerotomy sites after 23-gauge microincisional vitrectomy surgery (MIVS). METHODS: This is a retrospective study comprised of 185 eyes from 185 consecutive patients who underwent 23-gauge MIVS. The authors investigated the incidence and the clinical factors associated with conjunctival pigmentation, including indication of vitrectomy and surgical procedures. RESULTS: Eight eyes (4.3%) from 185 eyes showed conjunctival pigmentation after surgery within 7 days postoperatively. Light microscopic examination of excised tissue showed infiltration of abundant dark brownish colored melanophages with surrounding scattered, finely granular, melanin pigments. C3F8 gas tamponade (OR, 9.4; 95% CI, 1.9-44.5; P=0.005) was significantly associated with conjunctival pigmentation. The mean surface area (21.26±2.21 mm(2)) of three eyes with C3F8 tamponade was significantly larger than that of five eyes (2.51±0.73 mm(2)) without tamponade (P=0.025). Neither leakage nor hypotony was found. CONCLUSIONS: Intraocular melanin pigments could prolapse through the sclerotomy site and cause conjunctival pigmentation after 23-gauge MIVS. Thus, preoperative warning is necessary in patients with 23-gauge MIVS.

Combined 23-gauge microincisonal vitrectomy surgery and phacoemulsification with AcrySof toric intraocular lens implantation: a comparative study.

AIM: To compare AcrySof toric intraocular lens (IOL) and non-toric IOL in patients who had combined 23-gauge microincisional vitrectomy surgery (MIVS) and phacoemulsification for vitreoretinal diseases and cataract with pre-existing corneal astigmatism. METHODS: This is a prospective comparative study comprised of 30 patients (30 eyes) who had combined 23-gauge MIVS and phacoemulsification for vitreoretinal diseases and cataract with pre-existing regular corneal astigmatism greater than 1 diopters (D). In all, 15 eyes had AcrySof toric IOL (Alcon Laboratories) and 15 eyes had non-toric IOL (Akreos AO MI60; Bausch & Lomb) implantation. Main outcome measures were uncorrected visual acuity (UCVA), refractive cylinder, surgically induced astigmatism (SIA), and IOL misalignment during 6 months. RESULTS: The mean UCVA of the toric IOL group was better than the non-toric IOL group at postoperative months 1, 3, and 6 (P<0.001, respectively). The mean absolute residual refractive cylinder of the toric IOL group at postoperative week 1, and months 1, 3, and 6 was less than the non-toric IOL group (P=0.008, <0.001, <0.001, and <0.001, respectively). There was no difference in the mean SIA between the two groups (P>0.05, respectively). The mean toric IOL axis rotation was 3.52 ± 2.75°, which was within 5° in 66.7% of the toric IOL group and within 10° in 100%. CONCLUSIONS: Combined 23-gauge MIVS and phacoemulsification with AcrySof toric IOL implantation is an effective method of correcting vitreoretinal diseases and cataract and pre-existing corneal astigmatism, and the toric IOL showed good rotational stability, even in vitrectomized eyes for 6 months.

The management of the orthopaedic sequelae of meningococcal septicaemia: patients treated to skeletal maturity.

Septicaemia resulting from meningococcal infection is a devastating illness affecting children. Those who survive can develop late orthopaedic sequelae from growth plate arrests, with resultant complex deformities. Our aim in this study was to review the case histories of a series of patients with late orthopaedic sequelae, all treated by the senior author (CFB). We also describe a treatment strategy to address the multiple deformities that may occur in these patients. Between 1997 and 2009, ten patients (seven girls and three boys) were treated for late orthopaedic sequelae following meningococcal septicaemia. All had involvement of the lower limbs, and one also had involvement of the upper limbs. Each patient had a median of three operations (one to nine). Methods of treatment included a combination of angular deformity correction, limb lengthening and epiphysiodesis. All patients were skeletally mature at the final follow-up. One patient with bilateral below-knee amputations had satisfactory correction of her right amputation stump deformity, and has complete ablation of both her proximal tibial growth plates. In eight patients length discrepancy in the lower limb was corrected to within 1 cm, with normalisation of the mechanical axis of the lower limb. Meningococcal septicaemia can lead to late orthopaedic sequelae due to growth plate arrests. Central growth plate arrests lead to limb-length discrepancy and the need for lengthening procedures, and peripheral growth plate arrests lead to angular deformities requiring corrective osteotomies and ablation of the damaged physis. In addition, limb amputations may be necessary and there may be altered growth of the stump requiring further surgery. Long-term follow-up of these patients is essential to recognise and treat any recurrence of deformity.

Outcomes of endoscopic transpapillary gallbladder stenting for symptomatic gallbladder diseases: a multicenter prospective follow-up study.

BACKGROUND AND STUDY AIMS: Endoscopic transpapillary gallbladder drainage for symptomatic gallbladder disease is a safe and effective bridge therapy in patients at high risk for surgery or who have terminal liver disease and are awaiting transplantation. However, there are few reports on long-term results in terms of stent patency and clinical course. Our study was designed to investigate the long-term patency and clinical course after endoscopic transpapillary gallbladder stenting (ETGS) in patients with symptomatic gallbladder disease. PATIENTS AND METHODS: A total of 29 patients who were unsuitable for cholecystectomy underwent ETGS from June 2006 to March 2010 using a 7-Fr double-pigtail stent between the gallbladder and the duodenum. Their clinical progress, adverse events, and stent patency after ETGS were recorded prospectively in two tertiary referral centers. RESULTS: Technically, ETGS was successful in 23 (79.3%) of the 29 patients. The mean procedure time was 22.4 ± 11.5 min. Postprocedure adverse events were mild pancreatitis (8.7%) and cholestasis (8.7%), all of which resolved with conservative management. During the follow-up period (median 586 days, range 11-1403 days), 20 patients were analyzed as per protocol, and scheduled follow-up was performed. Late adverse events developed in four patients (20%), including distal migration (n = 2), cholangitis (n = 1), and recurrent biliary pain (n = 1). The remaining 16 patients were followed for more than 12 months (nine patients were followed for more than 24 months). Median stent patency was 760 days, as determined by the Kaplan-Meier method. CONCLUSIONS: As a primary therapy, ETGS is technically feasible and effective in patients who are unsuitable for cholecystectomy. ETGS may also provide long-term stent patency without the need for scheduled stent exchanges.

Screening of sepsis using leukocyte cell population data from the Coulter automatic blood cell analyzer DxH800.

INTRODUCTION: We determined the utility of leukocyte cell population data (CPD) for the screening of sepsis and fungemia. METHODS: Blood culture-positive CBC samples, 117 bacteremia and 27 fungemia, and 134 CBC samples from healthy controls were analyzed using the DxH800 and CPD of neutrophils, lymphocytes, and monocytes were analyzed. Immature granulocytes (IG) were counted using Sysmex XE-2100. RESULTS: The neutrophils and monocytes volume were increased significantly, and the neutrophils light scattering values were reduced significantly in the sepsis samples. ROC curves evidenced excellent sensitivity in the lymphocyte SD parameters (sensitivity 78-89%, specificity 78-87%), monocytes volume (at 177.5, sensitivity 88.2% specificity 87.3%), and monocytes volume SD (at 22.16, sensitivity 93.1% specificity 91.0%) for sepsis. The IG value was significantly higher in sepsis and the ROC curve evidenced a sensitivity of 82.8% and a specificity of 90.8% for sepsis. Only lower angle light scatter of lymphocytes SD value evidenced good sensitivity and specificity in the discrimination of fungemia from bacteremia (sensitivity 74.1%, specificity 72.4% at 12.6). CONCLUSION: Many of the leukocyte CPD have been identified as useful parameters of sepsis. Hopefully, these parameters can ultimately be incorporated into a decision rule for the screening of sepsis samples and to discriminate fungemia from bacteremia.

Subungual glomus tumours of the hand: diagnosis and outcome of the transungual approach.

We report 17 patients with a subungual glomus tumour. All complained of pain and tenderness when touched, and nine patients experienced severe pain in the cold. A transungual approach with nail plate avulsion on one side was used in all cases. A surgical microscope was used to localise and dissect the tumour and to repair the nail bed and matrix. This method has produced good results, without local recurrence or postoperative nail plate deformity.

Endoscopic ultrasound-guided versus conventional transmural drainage for pancreatic pseudocysts: a prospective randomized trial.

BACKGROUND AND STUDY AIMS: Although endoscopic ultrasound (EUS)-guided transmural drainage (EUD) is preferred over conventional transmural drainage (CTD) of pancreatic pseudocysts by endoscopy in many centers, its superiority with respect to technical success and clinical outcome has not yet been demonstrated in a prospective randomized trial. We conducted this prospective randomized trial to compare the technical success and clinical outcomes of EUD and CTD in treating pancreatic pseudocysts. PATIENTS AND METHODS: A total of 60 consecutive patients with pancreatic pseudocysts were randomly divided into two groups to undergo either EUD (n = 31) or CTD (n = 29) of pancreatic pseudocysts. The technical success rate, complications, and short-term and long-term results were prospectively evaluated. RESULTS: The rate of technical success of the drainage was higher for EUD (94 %, 29/31) than for CTD (72 %, 21/29; P = 0.039) in intention-to-treat analysis. In cases where CTD failed (n = 8), because the pseudocysts were nonbulging, a crossover was made to EUD, which was successfully performed in all these patients. Complications occurred in 7 % of the EUD and 10 % of the CTD group ( P = 0.67). During short-term follow-up, pseudocyst resolution was achieved in 97 % (28/29) in the EUD group and in 91 % (19/21) in the CTD group ( P = 0.565). Long-term results analyzed on a per-protocol basis showed no significant difference in clinical outcomes between EUD (89 %, 33/37) and CTD (86 %, 18/21, P = 0.696). CONCLUSIONS: We found that EUD and CTD can both be considered first-line methods of endoscopic transmural drainage of bulging pseudocysts, whereas EUD should be preferred for nonbulging pseudocysts.

A randomised phase II study of modified FOLFIRI.3 vs modified FOLFOX as second-line therapy in patients with gemcitabine-refractory advanced pancreatic cancer.

BACKGROUND: Only a few clinical trials have been conducted in patients with advanced pancreatic cancer after failure of first-line gemcitabine-based chemotherapy. Therefore, there is no current consensus on the treatment of these patients. We conducted a randomised phase II study of the modified FOLFIRI.3 (mFOLFIRI.3; a regimen combining 5-fluorouracil (5-FU), folinic acid, and irinotecan) and modified FOLFOX (mFOLFOX; a regimen combining folinic acid, 5-FU, and oxaliplatin) regimens as second-line treatments in patients with gemcitabine-refractory pancreatic cancer. METHODS: The primary end point was the 6-month overall survival rate. The mFOlFIRI.3 regimen consisted of irinotecan (70 mg m(-2); days 1 and 3), leucovorin (400 mg m(-2); day 1), and 5-FU (2000 mg m(-2); days 1 and 2) every 2 weeks. The mFOLFOX regimen was composed of oxaliplatin (85 mg m(-2); day 1), leucovorin (400 mg m(-2); day 1), and 5-FU (2000 mg m(-2); days 1 and 2) every 2 weeks. RESULTS: Sixty-one patients were randomised to mFOLFIRI.3 (n=31) or mFOLFOX (n=30) regimen. The six-month survival rates were 27% (95% confidence interval (CI)=13-46%) and 30% (95% CI=15-49%), respectively. The median overall survival periods were 16.6 and 14.9 weeks, respectively. Disease control was achieved in 23% (95% CI=10-42%) and 17% patients (95% CI=6-35%), respectively. The number of patients with at least one grade 3/4 toxicity was identical (11 patients, 38%) in both groups: neutropenia (7 patients under mFOLFIRI.3 regimen vs 6 patients under mFOLFOX regimen), asthaenia (1 vs 4), vomiting (3 in both), diarrhoea (2 vs 0), and mucositis (1 vs 2). CONCLUSION: Both mFOLFIRI.3 and mFOLFOX regimens were tolerated with manageable toxicity, offering modest activities as second-line treatments for patients with advanced pancreatic cancer, previously treated with gemcitabine.