Three candidate antisense target sites of mouse Fas gene were screened by PARASS (poly-A anchored RNA accessible sites screening) technology. They were target at Fas gene 297nt-317nt, 618nt- 638nt and 662nt-682nt. Antisense oligos (A1, A2 and A3) and DNAzymes (D1, D2, and D3) for every target site were designed and synthesized. In vitro, the validation of the sites were judged by antisense oligos included RNase H splicing and the DNAzyme degradation. The results indicated that A1, A2 and A3 introduced RNase H degradation. DNAzymes D1, D2 and D3 cleaved Fas mRNA effectively. Neither degradation observed in antisense oligo RNase H group in non-target site (1211-1231nt) and 2 bases mismatched of A3, nor splicing occurred in DNzyme group in non-target site ( 1211-1231nt) and 2 bases mismatched of D3. Site 2 and 3 were at the same positions with those of ISIS Pharmaceuticals. The effective antisense oligos and DNAzymes for Fas gene could be used for the research subsequently.

Objective To compare the outcomes of percutaneous pedicle instrumentation combined with vertebral augmentation or vertebra pedicle instrumentation for treatment of osteoporotic thoracolumbar fractures (OVCF) in elderly patients.Methods A retrospective case cohort study was conducted on 62 patients with OVCF manifesting non-neurological symptoms treated from January 2009 to January 2012.There were 22 males and 30 females,with a mean age of 61.3 years (range,55 to 70 years).Fracture level was T11 in 8 patients,T12in 20,L1 in 22 and L2 in 12.Treatments included percutaneous pedicle instrumentation combined with vertebral fracture fixation in 36 patients (Group A) and percutaneous pedicle instrumentation combined with vertebral augmentation in 26 patients (Group B).Operation time,intraoperative blood loss,anterior vertebral body height,sagittal Cobb angle and visual analogue score (VAS) were compared between the two groups.Results All patients were followed up for average 46.5 months (range,36 to 58 months).Operation time in Group A [(82.6 ±16.2) min] was shorter than that in Group B [(96.8 ± 20.6) min] (P ＜ 0.05).Blood loss in Group B [(40.5 ± 10.2) ml] was less than that in Group A [(52.2 ± 15.5) ml] (P ＜ 0.05).Before operation and 3 days and 1 year after operation,the anterior vertebral body height and sagittal Cobb angle in Group A showed no significant differences compared to Group B (all P ＞ 0.05).At the final follow-up,the ratio of anterior vertebral height and Cobb angle in Group B [(87.8 ± 2.5) ％,(7.8 ± 3.5) °] were better than these in Group A [(82.6 ±3.2)％,(9.1 ± 1.8)°] (P＜0.05).VAS showed no statistical significance between the two groups before and after operation (P ＞ 0.05).Bone cement leakage was seen in four patients in Group B.During the perioperative period,there were 3 patients with lung infection in Group A and 1 patient with lower limb deep vein thrombosis in Group B.No implant failure occurred in both groups.Conclusion Both procedures are effective in treating elderly patients with OVCF,but percutaneous pedicle instrumentation combined with vertebral augmentation is associated with better results in maintaining vertebral height and preventing kyphosis.

BACKGROUNDSevoflurane and propofol are widely used anesthetics for surgery. Studies on the mechanisms of general anesthesia have focused on changes in protein expression properties and membrane lipid. MicroRNAs (miRNAs) regulate neural function by altering protein expression. We hypothesize that sevoflurane and propofol affect miRNA expression profiles in the brain, expect to understand the mechanism of anesthetic agents.

METHODSRats were randomly assigned to a 2% sevoflurane group, 600 μg·kg - 1·min - 1 propofol group, and a control group without anesthesia (n = 4, respectively). Treatment group was under anesthesia for 6 h, and all rats breathed spontaneously with continuous monitoring of respiration and blood gases. Changes in rat cortex miRNA expression profiles were analyzed by miRNA microarrays and validated by quantitative real-time polymerase chain reaction (qRT-PCR). Differential expression of miRNA using qRT-PCR among the control, sevoflurane, and propofol groups were compared using one-way analysis of variance (ANOVA).

RESULTSOf 677 preloaded rat miRNAs, the microarray detected the expression of 277 miRNAs in rat cortex (40.9%), of which 9 were regulated by propofol and (or) sevoflurane. Expression levels of three miRNAs (rno-miR-339-3p, rno-miR-448, rno-miR-466b-1FNx01) were significantly increased following sevoflurane and six (rno-miR-339-3p, rno-miR-347, rno-miR-378FNx01, rno-miR-412FNx01, rno-miR-702-3p, and rno-miR-7a-2FNx01) following propofol. Three miRNAs (rno-miR-466b-1FNx01, rno-miR-3584-5p and rno-miR-702-3p) were differentially expressed by the two anesthetic treatment groups.

CONCLUSIONSSevoflurane and propofol anesthesia induced distinct changes in brain miRNA expression patterns, suggesting differential regulation of protein expression. Determining the targets of these differentially expressed miRNAs may help reveal both the common and agent-specific actions of anesthetics on neurological and physiological function.

Anesthesia, General ; Animals ; Brain ; drug effects ; metabolism ; Male ; Methyl Ethers ; pharmacology ; MicroRNAs ; genetics ; Propofol ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Real-Time Polymerase Chain Reaction

OBJECTIVETo investigate the associations between periodontal diseases, presence of Porphyromonas gingivalis (P. gingivalis), serum levels of pro-inflammatory mediators and preterm low birth weight (PLBW).

METHODS60 women (30 PLBW and 30 healthy women), were recruited after postpartum within 3 days in this case-control study. Periodontal measurements including plaque index (PI), bleeding index (BI), probing pocket depth (PD) and clinical attachment loss (CAL) were assessed. The subgingival plaque was collected before periodontal examination and analysed by polymerase chain reaction (PCR) for the presence of the 16S rRNA gene specific to P. gingivalis, while the venous and umbilical cord blood specimens collected were determined by enzyme-linked immunosorbent assay (ELISA).

RESULTSThe PLBW cases had a poorer oral conditions and the presence of P. gingivalis was found in a higher proportion in the PLBW than the healthy pregnant (56.7% vs. 30.0%, P < 0.05). Both of the presence of periodontitis and P. gingivalis have been found to be associated weakly with a shorter gestational age and a lower birth weight (P < 0.05). The levels of IL-1beta, IL-6 and prostaglandin-E2 (PGE2) in both of the blood samples showed higer levels between the PLBW and normal groups (P < 0.01). The IL-1beta and PGE2 levels in maternal serum were higher with a severe periodontal disease in the PLBW group (P < 0.01).

CONCLUSIONThere may be a possible link between periodontal diseases and PLBW.

Adult ; Case-Control Studies ; Dental Plaque ; Dental Plaque Index ; Female ; Humans ; Infant, Low Birth Weight ; Infant, Newborn ; Interleukin-6 ; Periodontal Diseases ; Periodontitis ; Porphyromonas gingivalis ; Pregnancy ; Premature Birth ; RNA, Ribosomal, 16S

BACKGROUNDCollapsin response mediator protein-2 (CRMP2), a multifunctional cytosolic protein highly expressed in the brain, is degraded by calpain following traumatic brain injury (TBI), possibly inhibiting posttraumatic neurite regeneration. Lipid peroxidation (LP) is involved in triggering postinjury CRMP2 proteolysis. We examined the hypothesis that propofol could attenuate LP, calpain-induced CRMP2 degradation, and brain injury after TBI.

METHODSA unilateral moderate controlled cortical impact injury was induced in adult male Sprague-Dawley rats. The animals were randomly divided into seven groups: Sham control group, TBI group, TBI + propofol groups (including propofol 1 h, 2 h, and 4 h groups), TBI + U83836E group and TBI + fat emulsion group. The LP inhibitor U83836E was used as a control to identify that antioxidation partially accounts for the potential neuroprotective effects of propofol. The solvent of propofol, fat emulsion, was used as the vehicle control. Ipsilateral cortex tissues were harvested at 24 h post-TBI. Immunofluorescent staining, Western blot analysis, and terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling were used to evaluate LP, calpain activity, CRMP2 proteolysis and programmed cell death. The data were statistically analyzed using one-way analysis of variance and a paired t-test.

RESULTSPropofol and U83836E significantly ameliorated the CRMP2 proteolysis. In addition, both propofol and U83836E significantly decreased the ratio of 145-kDa αII-spectrin breakdown products to intact 270-kDa spectrin, the 4-hydroxynonenal expression and programmed cell death in the pericontusional cortex at 24 h after TBI. There was no difference between the TBI group and the fat emulsion group.

CONCLUSIONSThese results demonstrate that propofol postconditioning alleviates calpain-mediated CRMP2 proteolysis and provides neuroprotective effects following moderate TBI potentially by counteracting LP and reducing calpain activation.

Animals ; Blotting, Western ; Brain Injuries ; drug therapy ; metabolism ; Calpain ; metabolism ; Intercellular Signaling Peptides and Proteins ; metabolism ; Lipid Peroxidation ; drug effects ; Male ; Nerve Tissue Proteins ; metabolism ; Propofol ; therapeutic use ; Proteolysis ; drug effects ; Rats ; Rats, Sprague-Dawley

Objective To investigate the mRNA expression of liver receptor homolog 1 (LRH-1) gene in patients with cholesterol gallstone disease so that to elucidate the biomolecular pathogenesis of gallstone for- mation.Methods Twenty-seven patients with cholesterol gallstone (CGS) and 14 controls were included in this study.Biliary composition was assayed and mRNA expression of hepatic LRH 1 gene was determined by real time polymorphism chain reaction.Results In CGS patients,expression of LRH-1 was significantly higher than that in controls (14.18?9.37 vs 7.86?6.19,P＜0.05),and cholesterol of bile was oversaturated (1.17?0.27).Conclusion The formation of CGS may be related to increased expression of hepatic LRH-1 gene.

OBJECTIVETo analyze the clinical and molecular cytogenetic features of hematologic malignancies with idic(20q-).

METHODSThe clinical data of 10 patients with idic (20q-) were analyzed. Karyotyping analysis was carried out with R banding technique. A CEP20 probe was used to perform single-color fluorescence in situ hybridization (FISH). A subtelomeric probe for 20q and a locus-specific probe for 20q12 were used to perform dual-color FISH. The literatures of hematologic malignancies with idic(20q-) were reviewed.

RESULTSOf the 10 cases, 2 were diagnosed as acute erythroid leukemia, 1 primary myelofibrosis, 3 myelodysplastic syndromes (MDS) and 4 highly suspected (HS-MDS). Karyotype analysis showed that one of the normal chromosome 20 allele was substituted by one or two metacentric isochromosomes smaller than the normal one in all 10 cases. It was confirmed to be der(20)del(20)(q11q13)idic(20)(p11), i.e., idic(20q-) by FISH assay. Partial cells in 2 of the 10 cases had 20q- as the sole karyotypic anomaly.

CONCLUSIONIdic(20q-) results from a pre-existing del(20q) and is strongly associated with MDS and acute erythroid leukemia. Idic(20q-) as a recurrent cytogenetic abnormality is helpful for diagnosing HS-MDS in patients with cytopenia but only slight or absent dysplasia.

Adult ; Aged ; Chromosome Aberrations ; Chromosome Deletion ; Chromosomes, Human, Pair 20 ; genetics ; Female ; Humans ; In Situ Hybridization, Fluorescence ; Isochromosomes ; Male ; Middle Aged ; Myelodysplastic Syndromes ; diagnosis ; genetics

OBJECTIVETo survey the status of diagnosis and treatment of biliary tract cancer in Shanghai.

METHODSA clinical epidemiology investigation was carried out on 658 new cases of biliary duct cancers aged 35-74, that registered between June 1997 and May 2001 in urban Shanghai. Clinical findings were collected in 390 gallbladder cancer, 195 bile duct cancer and 73 ampullary cancer.

RESULTSBiliary tract cancers mainly occurred in elderly patients. Ratio of males to female was 1:2.61 in gallbladder cancer, while bile duct cancer and ampullary cancer were slightly more common in men. Association with gallstones was 68.5%, 43.1% and 22.4% for gallbladder cancer, bile duct cancer and ampullary cancer, respectively. Diagnostic accuracy rate of B-ultrasonography was 63.1% in gallbladder cancer. Incidental gallbladder cancer accounted for 20%, while stage IVA and IVB patients reached up to 43.6%. Misdiagnosis rate was still high in bile duct cancer and ampullary cancer, it was 19.1% and 47.1% respectively. In addition, most patients presented jaundice at diagnosis. 69 cases (18.2%) of gallbladder cancer, 50 cases (25.6%) of bile duct cancer and 54 cases (74%) of ampullary cancer underwent radical resection, the 1-, 3- and 5-year survival rates were 58.5%, 42.8% and 40.7%, 58%, 28.3% and 11.1%, 81.5%, 39.2% and 26.9%, respectively. 79 patients with bile duct cancer underwent palliative drainage, and most cases died within 1 year. Metal endo-prostheses or plastic stents were placed into the biliary tract in 38 patients. The median survival was about 7 months.

CONCLUSIONSIt is difficult to make early diagnosis of biliary tract cancers. Standardization of the operation for gallbladder cancer must be respected. Surgical exploration should be undertaken when a bile duct cancer is suspected and there are no contraindications to surgery. Pancreatoduodenectomy should be recommended for ampullary cancer.

Biliary Tract Neoplasms ; diagnosis ; epidemiology ; mortality ; therapy ; China ; epidemiology ; Data Collection ; Female ; Humans ; Male ; Survival Rate

OBJECTIVETo distinguish trochlear calcification and intraorbital foreign body after eye injury in order to avoid misdiagnosis as well as mistreatment.

METHODSThe orbital CT images of 403 patients, who visited the Eye Hospital or the Second Affiliated Hospital of Wenzhou Medical College during May 2005-April 2007, were reviewed. The diagnosis of trochlear calcification and intraorbital foreign body was made together by a skilled radiologist as well as an ophthalmologist. General information and CT characteristics in the patients with trochlear calcification were collected.

RESULTSUsing CT scan images, 27 among 403 patients (6.69%) were identified with trochlear calcification. Three patients (3/27, 11.11%) were misdiagnosed by radiologists as intraorbital foreign body. Among the 27 patients with trochlear calcification, 23 (85.19%) were male and 4 (14.81%) were female, with an unilateral calcification in 7 patients (7/27, 25.93%) and bilateral in 20 (74.07%) . The highest occurrence of trochlear calcification was in 31-40 years old group (13/403, 3.23%) which reached to 12.87% (13/101) after age-correction. There were 3 types of trochlear calcification on the basis of CT images: commas, dot and inverted "U".

CONCLUSIONSThe trochlear calcification is not an uncommon phenomenon and should not be diagnosed as intraorbital foreign body, especially when it co-exists with eye injury in 31-40 years old group. Injury history and our classification method on the basis of CT images could help to avoid misdiagnosis.

Adolescent ; Adult ; Aged ; Calcinosis ; diagnostic imaging ; etiology ; Child ; Child, Preschool ; Eye Foreign Bodies ; diagnostic imaging ; etiology ; Eye Injuries ; complications ; diagnostic imaging ; Female ; Humans ; Infant ; Infant, Newborn ; Male ; Middle Aged ; Orbit ; diagnostic imaging ; Orbital Diseases ; diagnostic imaging ; etiology ; Tomography, X-Ray Computed

OBJECTIVETo evaluate the usefulness of multi-slice spiral CT (MSCT) in the post-operative assessment of cochlear implanted electrode.

METHODSTwenty-three cochlear implant recipients were enrolled in this study. All patients were examined with a SOMATOM Sensation 16-slice CT scanner (Siemens) using the following parameters: 120 kV, 100 mAs, 0. 75 mm collimation, 1 mm reconstruction slice thickness and increment, a pitch factor of 1, and a FOV of 100 mm. The axial images of interested ears were reconstructed with 0.1 mm increment and a FOV of 50 mm, and then volume rendering technique (VRT) reconstruction were done on the work station.

RESULTSThe electrode arrays were detected on axial CT images. Both inner ear and electrode array could be displayed on one image simultaneously. VRT provided an intuitionistic view of the relationship between electrode array and cochlea VRT showed the number of the electrode array in 20 patients implanted with Combi 40 + standard electrode array and demonstrated the shape, position, and insertion depth. The electrode array number determined by VRT was in accordance with the surgical findings in 18 patients, and was underestimated in two patients. In 3 patients with Combi 40 + compressed electrode array, only 4 to 5 electrodes arrays were clearly identified and others were not observed.

CONCLUSIONMSCT with VRT can provide useful three-dimensional information of the electrode array and indicate the exact relationship between electrode array and cochlea.

Adolescent ; Adult ; Child ; Child, Preschool ; Cochlear Implantation ; Cochlear Implants ; Ear, Inner ; diagnostic imaging ; Female ; Humans ; Imaging, Three-Dimensional ; Infant ; Male ; Middle Aged ; Postoperative Period ; Tomography, Spiral Computed