ObjectivesTo investigate the effects of 5-aza-2-deoxycytidine(5-Aza-dC),a methylation inhibitor,on the expression and methylation of tissue factor pathway Inhibitor (TFPI-2) gene in PANC1 cell line of pancreatic cancer.MethodsPANC1 cell lines were treated with different dosages of 5-Aza-dC ( 1× 10-7,5 × 10-7,1× 10 -6 mol/L).The status of TFPI-2 methylafion and expressions of TFPI-2 mRNA and protein were determined by MSP,RT-PCR,and Western blot.Results TFPI-2 gene CpG island was completely methylated,and there was no expression of TFPI-2 mRNA and protein without 5-Aza-dC treatment.After treatment with different dosages of 5-Aza-dC( 1 × 10-7,5 × 10 -7,1 × 10-6 mol/L),TFPI-2 gene CpG hypermethylation was reversed from incomplete methylated to complete non-methylated.The relative expressions of TFPI-2 mRNA were 0.211± 0.087,O.327 ± 0.068,0.609 ± 0.017; and the relative expressions of TFPI-2 protein were O.429 ± O.121,O.675 ± O.044,1.132 ± O.124 in a dose-dependent manner ( P ＜0.05 ).ConclusionsThe hypermethylatien of promoter region may be the primary reason for TFPI-2 gene expression down-rogtdation and inactivation.5-Aza-dC may reverse the hypermethylation of TFPI-2 gene,and induce the m-expression of TFPI-2 mRNA and protein.

Objective To compare clinical outcomes in cardiac transplant recipients treated with individualized dosing (ID) and fixed dosing (FD) of mycophenolate mofetil (MMF).Methods Fortyeight de novo cardiac transplant patients in ID group received MMF (2.0 g/day) in combination with calcineurin inhibitors and prednisone.The MMF dosages were adjusted individually based on clinical events and MPA trough levels (MPA-C0).The target range of C0 was maintained within 2.0-4.0rng/L.The FD group included 55 recipients retrospectively from the transplant database who were also treated with MMF (2.0 g/day).In this group,the MMF dose adjustment was performed empirically according to clinical events only.All of the follow-up data were collected up to 12 months post-transplantation.Results The follow-up rate was 95.8％ and the mean MPA-AUC0-12 was (54.37± 17.03) rng h-1 L-1 in the ID group.The mean MPA-C0 on the day 7 post-transplantation was significantly higher in the ID group than that at 12th month [(3.44 ± 0.58) mg/L vs.(2.79 ± 0.54)mg/L] (P＜0.05).The dose of MMF was significantly lower in the ID group at 4th week posttransplantation than in control group [(1.49± 0.48) g/day vs.(1.96 ± 0.39) g/d] (P＜0.05),while there was no significant difference at 12 th month post-transplantation [(1.61 ± 0.77) g/day vs.(1.68 ± 0.84) g/day] (P＞ 0.05).No significant difference was found in the incidence of acute rejection episode between two groups (8.7％ vs.9.1％,P＞0.05).57.6％ of overall side effects were observed within one month postoperatively,and the incidence of MPA-related side effects was significantly lower in the ID group than in the control group (47.8％ vs.67.3％,P＜0.05).Conclusion It was demonstrated that individualized use of MMF based on therapeutic drug monitoring may prevent MMF-related side effect and appears to be valuable to optimize the treatment of cardiac transplantation.

Objective To discuss the method of the surgical treatment for retained and regenerate hepaticolithiasis, and to improve the theraputic level. Methods In recent 5 years,in our hospital,136 cases of retained and regenerate hepaticolithiasis were treated by combination of hepatolobectomy with other operation, and these cases were analyzed retrospectivly. Results One patient died after operation (0. 7%),8 patients had other complications including: biliary fistuta(6 cases, 4.4%), subphrenic abscess (2 cases,1.5%). All cases were visited for 2.5 - 8.3 years, 115 (84.6%) cases had not any symptom, 6 (4.4%)cases had infection of biliary tract, 6(4.4%) cases had retained calculus, 9(6.6%) cases recurred calculus 2 -5 years after operation. Conclusion Retained and regenerate hepaticolithiasis should be treated by combination of hepatolobectomy with other operation and fiber choledochoscope and B ultrasound of operation, which can increase therapeutic effect.

Objective To evaluate the clinical effect of the intertrochanteric fractures with or without lateral femoral wall fractures using proximal femoral nail antirotation (PFNA).Methods From May 2008 to June 2011,102 patients with intertrochanteric fractures were treated with PFNA.In accordance with the preoperative three dimensional CT reconstruction(3D CT) images,the group A included 41 cases with lateral femoral wall fractures,and the other 61 cases with an intact lateral wall were in group B.According to the AO/OTA classification,there were 5 cases in 31-A2,36 in 31-A3 in group A,and 61 in 31-A2 in group B.The operative time,operative blood loss,average length of stay,postoperative X-ray images,and 3D CT images were collected for each patient.Time of partial weight-bearing,full weight-bearing and fracture healing were also recorded.Clinical evaluation was made using the functional recovery scale (FRS) of hip fractures.Results The mean operation time was 56±8 min in group A vs 45±6 min in group B; the mean blood loss was 238±21 ml vs 175±11 ml; the average length of stay was 17±3 days vs 15±3 days.On the postoperative radiography,the blowout of lateral trochanteric wall only occurred in 8 (19.5％,8/41) cases in group A and 3 (4.9％,3/61) in group B.According to the postoperative 3D CT,the similar findings were seen in 36 (87.8％.36/41) cases in group A and 45 (73.8％,45/61) in group B.Eighty-two cases were followed up for 6 to 35 months (mean.19.5 months).The mean FRS score was 64.2±4.8 points in group A and 76.5±7.9 points in group B.Conclusion When treating unstable intertrochanteric fractures,iatrogenic fractures in lateral trochanteric wall could be easily caused with using PFNA.3D CT could effectively evaluate iatrogenic trauma in the intertrochanteric fractures.

OBJECTIVETo investigate the correlation between trauma and pulmonary thromboembolism.

METHODSComminuted fractures and extensive soft-tissue contusion at both hind limbs were made by a falling weight from a height in 16 rabbits. Lung perfusion scanning was performed to obtain the radioactivity counts before trauma, at 1 h, 48 h and 96 h after trauma. All the data were divided into 4 groups based on the above 4 time points. The rabbits were sacrificed when positive findings on the pulmonary perfusion scanning appeared. Their lungs were harvested to be paraffin-embedded and stained with hematoxylin-erosin method for histological examination of thromboembolism. The randomized block design ANOVA and the method of least significant difference (LSD) were used for statistical analysis of the radioactivity counts.

RESULTSThe histological findings showed that pulmonary embolism developed in 6 of the 16 rabbits (37.5%). Five of the 6 pulmonary embolism rabbits presented neither clinical symptoms nor positive pulmonary embolism manifestations in the lung perfusion scanning. A significant difference was found in lung perfusion radioactivity between the pre-traumatic, post-traumatic 1h groups and post-traumatic 48 h and 96 h groups(P less than 0.05).

CONCLUSIONSFractures of the hind limbs accompanied with extensive soft-tissue contusion may cause pulmonary micro-embolism that is not sensitive to lung perfusion scanning and tends to have no clinical symptoms. Pulmonary embolism development may take more than two days after trauma.

Animals ; Female ; Fractures, Bone ; complications ; Male ; Pulmonary Embolism ; etiology ; Rabbits ; Wounds and Injuries ; complications

BACKGROUND: Micro-arc oxidation (MAO) treatment can improve the biological properties of titanium alloy implants. Previous studies mostly focused on the evaluation of titanium alloy plate, while the effects of the MAO-modified 3D titanium scaffold on the cell growth and differentiation were rarely reported. OBJECTIVE: To investigate the effect of MAO coating on the biological performance of cells seeded onto the 3D-printed porous titanium alloy scaffold. METHODS: Rat bone marrow mesenchymal stem cells were seeded onto the MAO-modified Ti6Al4V alloy scaffolds (experimental group) and unmodified scaffolds (control group). After 4 and 7 days of culture, cell/scaffold constructs were retrieved and processed for the assessment of cell morphology by using scanning electron microscopy, cytoskeletal staining analysis and cell viability assay were also evaluated. At 4, 7 and 11 days of culture, the levels of alkaline phosphatase and osteocalcin in the cell supernatant were detected. At 1, 4, 7 and 11 days of culture, the cell proliferation rate was measured using the MTT assay. RESULTS AND CONCLUSION: (1) At 4 and 7 days of culture, live/dead staining showed that the bone marrow mesenchymal stem cells grew well on the two kinds of scaffolds. The analysis of cytoskeleton staining showed that the cytoskeleton of the experimental group was stereo and polygonal, while the cells on the scaffold surface in the control group were flat and spindle-shaped, spreading along the macro structure of the scaffolds. Under the scanning electron microscopy, the cells in the experimental group arranged closely and spread in a good condition, with interconnected lamellipodia and filopodia that firmly adhered to the scaffold surface in an anchor-shaped structure; in the control group, less filopodia interconnected, less extracellular matrix, and flat and sheet-like cells were observed. (2) With the time increase, the levels of alkaline phosphatase and osteocalcin increased gradually in both groups. The alkaline phosphatase level in the experimental group was significantly higher than that in the control group at 7 and 11 days of culture (P < 0.05), while the osteocalcin level was higher in the experimental group than the control group at 11 days of culture (P < 0.05). (3) With the prolongation of culture time, the number of cells in the two groups increased gradually. The number of cells cultured in the experimental group was significantly higher than that in the control group at 7 and 11 days of culture (P < 0.05). To conclude, the MAO-modified titanium alloy scaffold is favorable for cell adhesion, proliferation and differentiation.

It is an urgent and difficult task to establish a simple and efficient method for identifying and isolating sperm cells from mixed stains in forensic science. Nucleic acid aptamers targeting sperm cell-surface proteins can be used for the separation and purification of sperms from mixed stain samples. Human lipocalin 6 (hLCN6) is an epididymal secreted protein that binds to the head and tail of sperm cells and is associated with sperm maturation. Using the systematic evolution of ligands by the exponential enrichment (SELEX) technique, magnetic bead-bound hLCN6 was used as the target molecule to screen for aptamers with high affinity and specificity to hLCN6 from a random single-stranded DNA (ssDNA) library. Through 15 rounds of positive selection and 3 rounds of negative selection, 24 clones were selected and subjected to sequence analysis. Subsequently, 4 candidate aptamers were selected and further examined for their binding affinity and specificity by enzyme-linked oligonucleotide adsorption (ELONA) and cell binding assays. One aptamer (H2) against hLCN6 with a high affinity and specificity was isolated and investigated by dot blotting and immunofluorescence staining. The result revealed that the candidate aptamer H2 with a dissociation constant of (3. 21 ± 0. 75) nmol/ L was able to recognize and specifically bind to hLCN6. The aptamer H2 also showed high affinity and specificity to human sperms in vitro, which establishes the foundation for the separation of sperm cells from mixed stain based on nucleic acid-protein interactions and provides a new scheme.

BACKGROUNDAs precise positioning of ankle radiography is not possible, quantitative measurement of all syndesmotic parameters on repeated ankle X-ray films may be of little value. The purpose of this study was to provide a set of scientific and objective evaluation criteria for assessing the quality of ankle fracture reduction accurately and reliably by an intelligent combining three-dimensional (3-D) computed tomography (CT) measurement model.

METHODSFrom June 2008 to March 2011, all the thin-slice CT images of 100 cases (50 males and 50 females) with normal ankle joint scanned by 16-slice spiral CT were collected. Two-dimensional (2-D) and 3-D images of ankle joints were generated by using multiple planar reconstruction (MPR) and surface shaded display (SSD) respectively. The relevant parameters about bone structures and their relationship were measured and analyzed based on 3-D topological narrow division technique and 3-D measurement techniques combining essential elements of point, line and surface.

RESULTSIn this study, the mean distance from lateral malleolus tip to talocrural articular surface, the tip of medial malleolus anterior colliculus to talocrural articular surface and lateral malleolus tip to the tip of medial malleolus anterior colliculus were (22.83 ± 1.12) mm, (12.84 ± 1.09) mm, and (61.18 ± 2.03) mm respectively in male group, and (20.16 ± 1.00) mm, (10.30 ± 1.05) mm and (53.00 ± 1.40) mm respectively in female group. The above three parameters were correlated with gender, height and weight (P < 0.05). However, the mean perpendicular distance from lateral malleolus tip to the plane through the tip of medial malleolus anterior colliculus, the talocrural angle, later clear space, medial clear space, and the superior clear space were (9.93 ± 0.29) mm, (10.01 ± 0.38)°, (1.94 ± 0.16) mm, (2.78 ± 0.19) mm, and (3.14 ± 0.15) mm respectively in 100 cases, were not significance correlated with gender, height and weight (P > 0.05).

CONCLUSIONSThis study could provide a certain amount of relevant data for the standard of injured ankle anatomical reduction and the second surgery planning after malunion. The methods of measurement are reliable, reproducible, and easy to apply in practice.

Adult ; Aged ; Ankle Joint ; diagnostic imaging ; Female ; Humans ; Image Processing, Computer-Assisted ; methods ; Male ; Middle Aged ; Tomography, X-Ray Computed ; methods ; Young Adult

OBJECTIVETo screen the molecular markers of DNA methylation with potential diagnostic value, and to explore their methylation features in Chinese colorectal neoplasma in order to find out ones with higher diagnostic value.

METHODSTissue samples of colorectal cancer and normal adjacent mucosa(>10 cm distance to tumors) from 10 colorectal cancer patients undergoing operation, and tissue samples of colorectal adenoma from 10 patients undergoing endoscopic resection in our center from June to August 2013 were collected respectively. Methylation status of 8 genes, such as SNCA, MAL, INA, SPG20, FBN1, CNRIP1, TFPI2, OSMR, was detected by BSP and qMSP to screen genes with potential diagnostic valua. ROC curve was drawn to analyze its diagnostic value.

RESULTSBSP measurement showed that the rate of DNA methylation of SNCA, SPG20 and FBN1 was 100% in colorectal cancer and adenoma, while no methylation was found in normal adjacent mucosa. The other 5 genes expressed in different extent in cancer, adenoma and normal adjacent mucosa. Among 10 cancer tissues and normal adjacent mucosa detected by qMSP method, positive SNCA methylation was found in 5 cases and 1 case respectively; positive SPG20 in 8 cases and 1 case respectively; positive FBN1 in 7 cases and 0 cases respectively, whose differences were significant (P=0.070, P=0.003 and P=0.007). The area under curve(AUC) of SNCA, SPG20, and FBN1 methylation for diagnosing colorectal cancer was 0.890, 0.730 and 0.880 respectively.

CONCLUSIONSNCA, SPG20 and FBN1 are potential genes with screening value for colorectal neoplasma.

Objective: To investigate the pathogen spectrum distribution and drug resistance of febrile neutropenic patients with hematological diseases in Shanghai. Methods: A retrospective study was conducted on the clinical isolates from the febrile neutropenic patients hospitalized in the departments of hematology in 12 general hospitals in Shanghai from January 2012 to December 2014. The drug susceptibility test was carried out by Kirby-Bauer method. WHONET 5.6 software was used to analyze pathogenic bacteria and drug susceptibility data. Results: A total of 1 260 clinical isolates were collected from the febrile neutropenic patients. Gram-positive bacteria accounted for 33.3% and Gram-negative bacteria accounted for 66.7%. Klebsiella pneumoniae (12.5%) , Stenotrophomonas maltophilia (9.5%) , Escherichia coli (9.1%) , Pseudomonas aeruginosa (8.7%) , Acinetobacter baumannii (6.6%) , Staphylococcus aureus (5.6%) and Enterococcus faecium (5.0%) were ranked in the first 7 of all pathogens. In the respiratory tract secretions specimens, non-fermented strains accounted for 56.2%. Stenotrophomonas maltophilia accounted for 15.2%. Enterobacteriaceae and coagulase-negative Staphylococci accounted for 42.3% (104/246) and 32.6% (85/246) respectively in blood samples. Enterobacteriaceae and Enterococcus bacteria accounted for 39.4% (76/193) and 28.5% (55/193) respectively in pus specimens. The detection rates of methicillin resistant Staphylococcus aureus (MRSA) and methicillin resistant coagulase negative Staphylococci (MRCNS) were 54.3% and 82.5%, respectively. Staphylococcus bacterial strain was not found to be resistant to linezolid, vancomycin and teicoplanin. The detection rate of Enterococcus vancomycin-resistant strains was 8.9%. Enterococcus was not detected resistance to oxazolidinone strains. Enterobacteriaceae bacteria were highly sensitive to carbapenems. The resistance rate of Pseudomonas aeruginosa to imipenem and meropenem was 34.1% and 15.8%, respectively. Stenotrophomonas maltophilia was more sensitive to minocycline hydrochloride, levofloxacin and sulfamethoxazole. The resistance rate of Acinetobacter baumannii only to cefoperazone-sulbactam was less than 10.0%. The antibiotic resistance rate of Klebsiella pneumoniae, Stenotrophomonas maltophilia, Pseudomonas aeruginosa and Acinetobacter baumanii to most of common antibiotics was lower than that of the CHINET surveillance. Conclusions The pathogenic strain distribution in common infection sites of febrile neutropenic patients was characterized. Bacterial resistance surveillance was better than the CHINET nationwide large sample surveillance in China.