Gli2 protein expression level is a feasible marker of ligand-dependent hedgehog activation in pancreatic neoplasms.

The hedgehog pathway is known to promote proliferation of pancreatic ductal adenocarcinoma (PDA) and has been shown to restrain tumor progression. To understand how hedgehog causes these effects, we sought to carefully examine protein expression of hedgehog signaling components during different tumor stages. Genetically engineered mice, Pdx1-Cre;LSL-KrasG12D and Pdx1-Cre;LSL-KrasG12D;p53lox/+, were utilized to model distinct phases of tumorigenesis, pancreatic intraepithelial neoplasm (PanIN) and PDA. Human pancreatic specimens of intraductal papillary mucinous neoplasm (IPMN) and PDA were also employed. PanIN and IPMN lesions highly express Sonic Hedgehog, at a level that is slightly higher than that observed in PDA. GLI2 protein is also expressed in both PanIN/IPMN and PDA. Although there was no difference in the nuclear staining, the cytoplasmic GLI2 level in PDA was modest in comparison to that in PanIN/IPMN. Hedgehog interacting protein was strongly expressed in the precursors, whereas the level in PDA was significantly attenuated. There were no differences in expression of Patched1 at early and late stages. Finally, a strong correlation between Sonic Hedgehog and GLI2 staining was found in both human and murine pancreatic tumors. The results indicate that the GLI2 protein level could serve as a feasible marker of ligand-dependent hedgehog activation in pancreatic neoplasms.

The production of coagulation factor VII by adipocytes is enhanced by tumor necrosis factor-α or isoproterenol.

BACKGROUND: A relationship has been reported between blood concentrations of coagulation factor VII (FVII) and obesity. In addition to its role in coagulation, FVII has been shown to inhibit insulin signals in adipocytes. However, the production of FVII by adipocytes remains unclear. OBJECTIVE: We herein investigated the production and secretion of FVII by adipocytes, especially in relation to obesity-related conditions including adipose inflammation and sympathetic nerve activation. METHODS: C57Bl/6J mice were fed a low- or high-fat diet and the expression of FVII messenger RNA (mRNA) was then examined in adipose tissue. 3T3-L1 cells were used as an adipocyte model for in vitro experiments in which these cells were treated with tumor necrosis factor-α (TNF-α) or isoproterenol. The expression and secretion of FVII were assessed by quantitative real-time PCR, Western blotting and enzyme-linked immunosorbent assays. RESULTS: The expression of FVII mRNA in the adipose tissue of mice fed with high-fat diet was significantly higher than that in mice fed with low-fat diet. Expression of the FVII gene and protein was induced during adipogenesis and maintained in mature adipocytes. The expression and secretion of FVII mRNA were increased in the culture medium of 3T3-L1 adipocytes treated with TNF-α, and these effects were blocked when these cells were exposed to inhibitors of mitogen-activated kinases or NF-κB activation. The ß-adrenoceptor agonist isoproterenol stimulated the secretion of FVII from mature adipocytes via the cyclic AMP/protein kinase A pathway. Blockade of secreted FVII with the anti-FVII antibody did not affect the phosphorylation of Akt in the isoproterenol-stimulated adipocytes. CONCLUSION: Obese adipose tissue produced FVII. The production and secretion of FVII by adipocytes was enhanced by TNF-α or isoproterenol via different mechanisms. These results indicate that FVII is an adipokine that plays an important role in the pathogenesis of obesity.

microRNA-18a induces apoptosis in colon cancer cells via the autophagolysosomal degradation of oncogenic heterogeneous nuclear ribonucleoprotein A1.

It is well known that microRNAs (miRs) are abnormally expressed in various cancers and target the messenger RNAs (mRNAs) of cancer-associated genes. While (miRs) are abnormally expressed in various cancers, whether miRs directly target oncogenic proteins is unknown. The present study investigated the inhibitory effects of miR-18a on colon cancer progression, which was considered to be mediated through its direct binding and degradation of heterogeneous nuclear ribonucleoprotein A1 (hnRNP A1). An MTT assay and xenograft model demonstrated that the transfection of miR-18a induced apoptosis in SW620 cells. A binding assay revealed direct binding between miR-18a and hnRNP A1 in the cytoplasm of SW620 cells, which inhibited the oncogenic functions of hnRNP A1. A competitor RNA, which included the complementary sequence of the region of the miR-18a-hnRNP A1 binding site, repressed the effects of miR-18a on the induction of cancer cell apoptosis. In vitro single and in vivo double isotope assays demonstrated that miR-18a induced the degradation of hnRNP A1. An immunocytochemical study of hnRNP A1 and LC3-II and the inhibition of autophagy by 3-methyladenine and ATG7, p62 and BAG3 siRNA showed that miR-18a and hnRNP A1 formed a complex that was degraded through the autophagolysosomal pathway. This is the first report showing a novel function of a miR in the autophagolysosomal degradation of an oncogenic protein resulting from the creation of a complex consisting of the miR and a RNA-binding protein, which suppressed cancer progression.

The diagnostic accuracy of high-resolution endoscopy, autofluorescence imaging and narrow-band imaging for differentially diagnosing colon adenoma.

BACKGROUND AND STUDY AIMS: Conventional colonoscopy can result in unnecessary biopsy or endoscopic resection due to its inability to distinguish adenomas from hyperplastic polyps. This study therefore evaluated the efficacy of high-resolution endoscopy (HRE), autofluorescence imaging (AFI), and narrow-band imaging (NBI) in discriminating colon adenoma from hyperplastic polyps. PATIENTS AND METHODS: This was a prospective multicenter study in patients undergoing AFI and NBI examinations. HRE, AFI, and NBI images were classified into two groups based on morphological characteristics, the predominant color intensities, and the visibility of meshed capillary vessels, respectively. Each of the endoscopic photographs were independently evaluated by a single endoscopist. The images were then assessed by three specialists and three residents, the latter having performed < 500 colonoscopies and < 30 NBI and AFI examinations. Diagnostic test statistics were calculated to compare the accuracy in differentiating colon adenoma from hyperplastic polyps for each method. RESULTS: A total of 183 patients were enrolled in the study and 339 adenomas and 85 hyperplastic polyps were identified. AFI and NBI could distinguish adenoma from hyperplastic polyps with an accuracy of 84.9 % and 88.4 %, respectively, whereas HRE exhibited an accuracy of 75.9 %. In the 358 lesions in which the AFI diagnosis was consistent with that of NBI, the accuracy, sensitivity, and specificity were high, at 91.9 %, 92.7 %, and 92.9 %, respectively. During the study comparing specialists and residents, AFI and NBI dramatically improved the diagnostic accuracy of residents from 69.1 % to 86.1 % and 84.7 %, respectively. CONCLUSIONS: Both AFI and NBI are considered to be feasible tools that can discriminate colon adenoma from hyperplastic polyps, and their use may be particularly beneficial for less-experienced endoscopists.

K-ras mutations and cell kinetics in Helicobacter pylori associated gastric intestinal metaplasia: a comparison before and after eradication in patients with chronic gastritis and gastric cancer.

BACKGROUND: Helicobacter pylori related gastric intestinal metaplasia (IM) is considered to be a precancerous lesion. AIMS: To identify the effects of H pylori eradication on K-ras mutations, cell kinetics in IM and histological changes in patients with and without gastric cancers in a one-year prospective study. METHODS: Patients included group A (n = 39), chronic gastritis, and group B (n = 53), intestinal-type early gastric cancer patients who had all undergone endoscopic mucosal resection (n = 25) or surgical resection (n = 28). K-ras codon 12 mutations in IM were examined, followed by DNA sequencing analysis. Proliferating and apoptotic cells were detected with anti-Ki-67 antibody and using the TUNEL method, respectively. RESULTS: The incidence of K-ras mutations in the cancer was only 3.8%. The mutant K-ras in IM was observed more frequently in group A (46.2%) than in group B patients (1.9%) (p<0.005). After eradication, the K-ras mutations significantly declined to 12.8% in group A (p<0.005). The mutation pattern of K-ras codon 12 before eradication was that GGT was mainly changed to AGT (50%) in group A. AGT transformation was not affected by treatment. Apoptosis in IM showed an increase after H pylori eradication in both groups (p<0.05 in group A) although no histological improvement in IM was observed. The monocyte score was significantly higher in group A than in group B (p<0.05); the score improved significantly after eradication. CONCLUSIONS: K-ras mutations in IM do not always play a role in gastric carcinogenesis but cell kinetics, especially apoptosis, in IM may contribute to it. There are early events in K-ras mutations which are influenced by H pylori infection; some mutations may also be selected by eradication. These unstable K-ras mutations in IM may be related to lymphocyte infiltration caused by H pylori infection.

Reduced expression of syndecan-1 affects metastatic potential and clinical outcome in patients with colorectal cancer.

Syndecan-1 is a transmembrane heparansulfate proteoglycan which regulates cell-to-cell or cell-to-extracellular matrix interactions and may influence malignant cell behavior. We investigated the alterations of syndecan-1 expressions in colorectal cancers and analyzed the relationship between histological and clinical characteristics. Syndecan-1 protein expression in colorectal cancer tissues was investigated with immunohistochemical staining of resected specimens. In situ hybridization was performed using syndecan-1 riboprobe to confirm the transcriptional signals. Syndecan-1 mRNA expression in cancer cell lines cultured with or without methylation inhibitor was also analyzed by quantitative PCR. Out of 105 specimens tested, less than 25% of tumor cells were stained with anti-syndecan-1 monoclonal antibody in 36 (34.3%). In situ hybridization showed a similar staining profile to that of immunohistochemistry. Syndecan-1 mRNA expression was increased by the methylation inhibitor 5-aza-2'-deoxycytidine, suggesting that the hypermethylation is involved in the suppression of syndecan-1 expression. Clinically, the incidence of metastasis to lymphnode or liver in patients with syndecan-1-negative tumors was significantly high. Among T1 colorectal cancers displaying a primary invasive phase, lymphnode metastasis, undifferentiated characters and 'budding' of cancer cells were more common in syndecan-1-negative tumors. The survival rate in patients with syndecan-1-negative tumors was decreased significantly in a stage-independent manner. These results suggest that the reduction of syndecan-1 expression in colorectal cancer cells, which is supposed to be regulated at the transcription level, is closely related to invasive character. The evaluation of syndecan-1 expression in colorectal cancer may allow prediction of patients' survival after surgery.

Posterior occipitocervical reconstruction using cervical pedicle screws and plate-rod systems.

STUDY DESIGN: This retrospective study was conducted to analyze the clinical results in 26 patients with lesions at the craniocervical junction that had been treated by occipitocervical reconstruction using pedicle screws in the cervical spine and occipitocervical rod systems. OBJECTIVES: To evaluate the effectiveness of pedicle screw fixation in occipitocervical reconstructive surgery and to introduce surgical techniques. SUMMARY OF BACKGROUND DATA: Many methods of occipitocervical reconstruction have been reported, but there have been no reports of occipitocervical reconstruction using pedicle screws and occipitocervical rod systems for reduction and fixation. METHODS: Twenty-six patients with lesions at the craniocervical junction underwent reconstructive surgery using pedicle screws in the cervical spine and occipitocervical rod systems. The occipitocervical lesions were atlantoaxial subluxation associated with basilar invagination, which was caused by rheumatoid arthritis in 19 patients and other disorders in 7. The lowest cervical vertebra of fusion in 16 patients was C2, and the remaining 10 patients underwent fusion downward from C3 to C7. Flexion deformity of the occipitoatlantoaxial complex was corrected by application of extensional force, and upward migration of the odontoid process was reduced by application of combined force of extension and distraction between the occiput and the cervical pedicle screws. RESULTS: Solid fusion was achieved in all patients except two with metastatic vertebral tumors who did not receive bone graft for fusion. Correction of malalignment at the craniocervical junction was adequate, and postoperative magnetic resonance imaging showed improvement of anterior compression of the medulla oblongata. There were no neurovascular complications of cervical pedicle screws. CONCLUSIONS: Occipitocervical reconstruction by the combination of cervical pedicle screws and occipitocervical rod systems provided the high fusion rate and sufficient correction of malalignment in the occipitoatlantoaxial region. Results of this study showed the effectiveness of cervical pedicle screw as a fixation anchor for occipitocervical reconstruction.

One-stage posterior decompression and reconstruction of the cervical spine by using pedicle screw fixation systems.

OBJECT: This retrospective study was conducted to analyze the results of one-stage posterior decompression and reconstruction of the cervical spine by using pedicle screw fixation systems in 46 patients. METHODS: Causes of cervical myelopathy in these 46 patients included spondylosis or ossification of the posterior longitudinal ligament, rheumatoid arthritis, metastatic or primary vertebral tumors, cervical spinal injuries, and spinal cord tumor. Thirty-three patients underwent this one-stage procedure as primary surgery. In the remaining 13 patients who had previously undergone laminectomies, the one-stage procedure was performed as salvage surgery. Cervical pedicle screws were inserted into the pedicles after probing and tapping. Graft bone was placed on the bilateral lateral masses, and pedicle screws were interconnected longitudinally by either plates or rods. Postoperatively, 26 patients showed improved neurological status (at least one grade improvement on Frankel's functional classification). There were no cases of neurological deterioration postoperatively. Solid bony fusion was obtained in all patients, except in seven patients with metastatic tumor who did not receive bone grafts. Correction of kyphosis was satisfactory. Postoperative radiological evaluation revealed that 10 (5.3%) of 190 screws inserted into the cervical vertebrae had perforated the cortex of the pedicles; however, no neurovascular complications were caused by the perforations. CONCLUSIONS: The pedicle screw fixation procedure, which does not require the lamina to be used as a stabilizing anchor, has proven to be valuable when performing one-stage posterior decompressive and reconstructive surgery in the cervical spine. The risk to neurovascular structures in this procedure, however, cannot be completely eliminated. Thorough knowledge of local anatomy and application of established surgical techniques are essential for this procedure.

Occipitoatlantal instability associated with articular tropism.

The authors report the case of a patient with rotatory posterior subluxation of the occiput on the atlas associated with tropism of the O-C1 articulations. Lateral flexion-extension plain films demonstrated 5 mm of posterior translation of the occiput on the atlas. Tomographs revealed tropism of the O-C1 articulations, and CT scans on extension demonstrated posterior rotatory subluxation of the occiput on the atlas. The patient had no osseous abnormality caudally below the atlantoaxial joint, and underwent posterior occipitoatlantal fusion by wiring. At the 4-year follow-up, the grafted bone had been incorporated, and the patient was completely free from preoperative symptoms. In the present patient, occipitoatlantal instability is presumed to have derived from articular tropism.

Small depressed neoplasms of the large bowel: radiographic visualization and clinical significance.

Small depressed and flat neoplasms (small flat adenomas) exist-in the large bowel and in the stomach. Most have been detected with colonoscopy. We tried to delineate these lesions radiographically. Radiographically, the contour of the lesions measuring 5 mm and less was visualized in 32 (60.4%) of 53 lesions. Radiographic examination preceded colonoscopy in 23 (71.9%) of these lesions. Ten (83.3%) of 12 lesions measuring 6-10 mm were visualized. The central depression was clearly delineated in 32 (60.4%) of 53 lesions measuring 5 mm and less and in 5 of the 12 (41.7%) lesions measuring 6-10 mm. The central depression was not demonstrated in all lesions whose contours were not visualized. The clinical significance of the small depressed neoplasm lies in the fact that a certain number may be a precursor of rapidly growing carcinoma with early submucosal invasion.

Anterior decompression and stabilization with the Kaneda device for thoracolumbar burst fractures associated with neurological deficits.

One hundred and fifty consecutive patients who had a burst fracture of the thoracolumbar spine and associated neurological deficits were managed with a single-stage anterior spinal decompression, strut-grafting, and Kaneda spinal instrumentation. At a mean of eight years (range, five years to twelve years and eleven months) after the operation, radiographs showed successful fusion of the injured spinal segment in 140 patients (93 per cent). Ten patients had a pseudarthrosis, and all were managed successfully with posterior spinal instrumentation and a posterolateral arthrodesis. The percentage of the canal that was obstructed, as measured on computed tomography, improved from a preoperative mean of 47 per cent (range, 24 to 92 per cent) to a postoperative mean of 2 per cent (range, 0 to 8 per cent). Despite breakage of the Kaneda device in nine patients, removal of the implant was not necessary in any patient. None of the patients had iatrogenic neurological deficits. After the anterior decompression, the neurological function of 142 (95 per cent) of the 150 patients improved by at least one grade, as measured with a modification of the grading scale of Frankel et al. Fifty-six (72 per cent) of the seventy-eight patients who had preoperative paralysis or dysfunction of the bladder recovered completely. One hundred and twenty-five (96 per cent) of the 130 patients who were employed before the injury returned to work after the operation, and 112 (86 per cent) of them returned to their previous job without restrictions. We concluded that anterior decompression, strut-grafting, and fixation with the Kaneda device in patients who had a burst fracture of the thoracolumbar spine and associated neurological deficits yielded good radiographic and functional results.

Reconstruction of an iliac crest defect with a bioactive ceramic prosthesis.

Between June 1987 and December 1990, an iliac crest prosthesis made of bioactive apatite- and wollastonite-containing glass ceramic (A-W.GC) was used in 60 patients for the reconstruction of the iliac crest defect after harvesting autogenous tricortical iliac bone graft. The clinical results of this prosthesis were satisfactory. No patients felt spontaneous pain in the reconstructed area, and 93% of the patients had no tenderness there. In the radiological evaluation at the final follow-up, no apparent "radiolucent clear zone" was detected at the prosthesis-iliac bone junction in 98% of the patients. Excellent new bone formation between the prosthesis and the iliac crest was also noticed in 96% of the patients. The A-W.GC iliac crest prosthesis was beneficial for reconstruction of the iliac crest defect.

The treatment of osteoporotic-posttraumatic vertebral collapse using the Kaneda device and a bioactive ceramic vertebral prosthesis.

Twenty-two patients with neurologic deficit due to delayed posttraumatic vertebral collapse after osteoporotic compression fractures of the thoracolumbar spine underwent anterior decompression and reconstruction with bioactive Apatite-Wollastonite containing glass ceramic vertebral prosthesis and Kaneda instrumentation. Eighteen patients previously had minor trauma that resulted in a mild vertebral compression fracture without any neurologic involvement and were either conservatively treated or not treated at all. Four had no history of back injury. The preoperative neurologic status was incomplete paralysis in all patients. The average age at surgery was 66 (53-79) years. The average follow-up was 34 (20-58) months after surgery. All patients had returned to their daily living with neurologic recovery and stable spine. This type of anterior procedure is effective in the osteoporotic patients and there was a very low incidence of instrumentation failure and very low morbidity.

Clinical study on stability of combined distraction and compression rod instrumentation with posterolateral fusion for unstable degenerative spondylolisthesis.

The authors examined the stability of combined distraction and compression rod instrumentation with posterolateral fusion in 40 consecutive patients with unstable degenerative spondylolisthesis. All operations were performed by floating fusion of L3-4 or L4-5 after decompression procedures. Mobility at the fused level was checked every 4 weeks after operation by the disc space angle on the functional radiographic films without brace. The average period of postoperative follow-up was 26 months. In 30 patients, no mobility was found at any time. In six patients, any mobility disappeared within 24 weeks, and in three patients, within 1 year. Pseudoarthrosis was found in one patient. The solid fusion rate was 97.5%. The values of percent slippage and slip angle were slightly improved. Lumbar lordosis was in the normal range at follow-up.

[Spinal epidural tumors without destructive bone change: clinical and radiological studies].

CT in 61 patients having spinal epidural tumors were reviewed. Of these patients twelve had an epidural mass lacking of destructive bone change. Histological examination of these 12 cases revealed that usual carcinomatous tumors are relatively less common but sarcomatous tumors and malignant hematologic neoplasms are more likely to form an epidural mass without bone destruction. The tumors preponderance thoracic spine and extended usually more than three spinal levels along spinal axis. In all but one tumor masses were found in the intervertebral foramen and the paraspinal region in addition to the spinal canal. Contrast enhanced CT was particularly effective to delineate tumor location and extension. Ring enhancement of the dural sac which constricted by epidural masses was considered pathognomonic of epidural tumors.