Angina Pectoris, Variant ; physiopathology ; Arrhythmias, Cardiac ; Electrocardiography ; Humans ; Male ; Middle Aged

BACKGROUND:At present, it is controversial to choose methotrexate therapy alone or triple therapy in the treatment of rheumatoid arthritis.
OBJECTIVE:To systemical y evaluate the efficacy and safety of methotrexate therapy and methopterin+hydroxychloroquine+sulfasalazine triple therapy in the treatment of rheumatoid arthritis.
METHODS:Computer retrieval was performed on the Cochrane library, PubMed, EMBASE NRR (http://www.updatesoftware.com/National), CCT (http://www.control ed-trails.com) and the Chinese biomedical literature database. Manual retrieval was performed on Chinese major orthopedic journals. Randomized control ed trials of methotrexate therapy alone and methopterin+hydroxychloroquine+sulfasalazine triple therapy treatment for rheumatoid arthritis were col ected. Methodological quality of the included studies was evaluated. The statistical software Revman 5.0 provided by the Cochrane col aboration was used.
RESULTS AND CONCLUSION: A total of four literatures in a randomized control ed trial, a total of 297 cases were included;the methodological quality was al class B. Meta-analysis results showed that blood sedimentation improvements were larger in methotrexate therapy alone than in the triple therapy [mean difference=7.01, 95%confidence interval (CI) (2.82, 11.19), P=0.001). Improvements of joints were better in triple therapy than that in single therapy [OR=0.62, 95%CI (0.41, 0.95), P=0.03). There was no significant difference in adverse events. Meta analysis could not be done in ACR, because literature did not provide detailed data. Compared with the triple therapy treatment, methotrexate therapy alone obviously reduced the sedimentation of rheumatoid arthritis in adults, but in terms of joint function improvement, triple therapy was superior to methotrexate therapy alone. Moreover, on the adverse event, there was no significant difference between the two groups. Because this system evaluation included a less number of cases, it stil needs strict large-sample randomized control ed studies to increase the strength of the evidence.

A magnetic extraction sorbent based on Fe3 O4@poly ( methacrylic acid-co-ethylene dimethacrylate) ( Fe3 O4@MAED ) was synthesized using methacrylic acid and ethylene dimethacrylate as monomer and cross-linker, respectively. The magnetic sorbent was characterized by FTIR spectroscopy, elemental analysis and transmission electron microscopy. At the same time, the Fe3 O4@MAED was used to extract four benzoylurea pesticides in water and juice samples under magnetic dispersive solid phase microextraction ( MDSPME ) format. To obtain the optimal extraction conditions, several parameters, including the amount of sorbent, desorption solvent, extraction and desorption time, pH value, and ionic strength in sample matrix were investigated. Based on this, a fast, simple and sensitive method for the determination of benzoylurea pesticides in water and juice samples was developed by the combination of MDSPME with HPLC equipped with diode array detector. Under the optimized experimental conditions, the developed method possessed expected linear dynamic ranges, coefficients of correlation ( R2>0. 99 ) and sensitivity. The limits of detection (S/N=3) for target analytes were 0. 10-0. 19 μg/L in water and 0. 12-0. 30 μg/L in juice, respectively. The RSDs for intra-day were less than 7% and inter-day RSDs were less than 11%. The developed method was successfully applied to the determination of benzoylurea pesticides residues in water and juice samples and the recoveries of spiked target compounds were in the range of 69. 4%-118. 0%. The results demonstrated that the Fe3 O4@ MAED could extract benzoylurea pesticides effectively through multi-interactions including hydrophobic, hydrogen-bond and ion-exchange interactions between sorbent and analytes.

Objective To explore the diagnosis and treatment of pancreatic duct stones. MethodsClinical data of 29 cases of pancreatic duct stones were analyzed retrospectively. ResultsChronic pancreatitis was complicated in 21 cases, cholelithiasis in 12, pancreatic cancer in 2 . The correct diagnosis rate of ultrosonography, CT, ERCP and MRCP was 89.7%(26/29), 65.0%( 13/20 ),100%(6/6) and 100%(4/4) respectively. Six cases underwent EST,16 cases did transpancreatic duct lithotomy plus Roux-en-Y pancreaticojejunostomy, Three cases did sphincteroplasty, pancreaticoduodenectomy and exploratory laparotomy were performed in one each. ConclusionsChronic panreatitis cholelithiasis is the main causes of pancreatic duct stones. Imaging examinations help to make diagnosis preoperatively. Transpancreatic duct lithotomy plus Roux-en-Y pancreaticojejunostomy is the therapy of choices for patients with pancreatic duct stones.

BACKGROUND: Anterior cervical decompression and bone graft fusion is always the standard operation type for the treatment of cervical spondylotic radiculopathy and myelopathy. However, secondary degeneration and new symptoms caused by the stress increase of adjacent segments following anterior cervical spinal fusion have been paid more and more attention. Artificial cervical disc replacement provides a new choice for the treatment of cervical syndrome.OBJECTIVE: To observe the therapeutic effect of BRYAN& artificial cervical disc displacement for the treatment of cervical spondylotic radiculopathy and myelopathy.DESIGN: Self-control experiment.SETTING: Department of Orthopaedics, Sixth Hospital of Ningbo.PARTICIPANTS: Seventeen patients with cervical spondylotic radiculopathy and/or myelopathy admitted to the Department of Orthopaedics, Sixth Hospital of Ningbo were involved between December 2003 and February 2005 were involved in the study. Informed consents were obtained, and all participates were approved by Medical Ethics Committee.There were 9 patients with cervical spondylotic myelopathy, and 8 patients with cervical spondylotic radiculopathy.BRYAN& artificial cervical disc system is an artificial cervical intervertebral disc with composite structure. It is constructed by two titanium alloy end plates and polymer core, which can bear loading and took polyisocyanate as matrix.METHODS: BRYAN& artificial cervical disc system was used in the treatment. All the patients underwent single-segment displacement. Two patients underwent the displacement at C3-4, five at C4-5, 8 at C5-6 and 2 at C6-7. At postoperative 1, 3and 6 months, X-ray lateral position, anteflexion position, posterior extension position, the left and right side dynamic position plains were taken to investigate prosthetic stability. Neurologic function was evaluated according to Japanese Orthopaedic Association (JOA) scoring system.MAIN OUTCOME MEASURES: ① Investigating whether or not prosthesis displaced by means of dynamic position plains.② Investigating the recovery of neurologic function by JOA scoring system.RESULTS: ①JOA average scores were elevated to 15 from preoperative 8.5. Nervous system symptoms had improved to different extents, and the mean improvement rate was 75%. At postoperative 1,3 and 6 months, all the patients were followed up and their X-ray plains of cervical lateral position and flexion-extension position showed good stability,indicating that motor function was not lost obviously. No heterotopic ossification was found around the prothesis. ②Wound infection was not found in any patient. At postoperative 3 months, less than 2 mm slight displacement appeared in the prothesis in one patient, and there were no obvious symptoms. Prothesis abrasion, subsidence, bone bridge formation and other rejections were not found in the other prothesis. Prothesis had good biocompatibility, and could be examined by routine CT and MRI.CONCLUSION: Artificial cervical disc replacement can obviously improve the symptoms of patients, and has good therapeutic effects in the treatment of cervical syndrome.

Objective:To construct lentiviral vector specific for mouse B7-1 RNA interference and study lentivirus-mediated B7-1 gene silencing effects in L929 fibroblast cells.Methods:Three candidate sequences for B7-1 RNAi selected from coding sequence of mouse B7-1 transcription were used to design short hairpin RNA ( shRNA ) templates and then cloned into lentiviral expression plasmid followed with correctness identification of inserted sequence by DNA sequencing.Recombinant lentivirus were prepared by co-transfecting lentiviral expression vector and packaging plasmids into 293T cells.Then the resulting culture supernatant containing infectious lentiviral particles was pooled and centrifuged via ultra-centrifugation.Infectious titer of the preparations was determined by detecting the expression of GFP in 293T cells after transfected by lentivirus.Cultured L929 cells were transfected with lentivirus to deter-mine transduction efficiency and silencing efficacy of B7-1 expression by flow cytometry.Transducted L929 cells were then screened using puromycin to generate stable cell clones followed by flow cytometry analysis of GFP and B7-1 expression.A mixed reaction system consisting of stable B7-1 silencing L929 cells and mouse splenic T cells was used to analyze ability of the established cell line to trigger T cells proliferation.Results: Lentiviral expression vector for mouse B7-1 RNAi was correctly constructed with inserted sequences as designed.Recombinant RNAi lentivirus were prepared with titers ranging (3-5) ×108 TU/ml and efficacy to mediate GFP transgene expression and B7-1 silencing.B7-1 expression and the ability to trigger T cells proliferation of stable L929 cells were suppressed significantly ( P<0.05 ).Conclusion: We generated lentiviral vector specific for mouse B7-1 RNAi with high performance of transduction efficiency as well as B7-1 silencing efficacy and the recombinant RNAi lentivirus can mediated stable B7-1 gene silencing in L929 cells and inhibition of T cells proliferation induced by B7-1/CD28 co-stimulatory signal.

Objective The effect of serum exosome isolation method on obtained exosomal RNA is not yet clear .The aim of this study was to provide evidences to selected exosome isolation methods .Methods This was a method comparison study to assess three methods .Vein blood samples were collected from 3 healthy donors from Nanfang hospital , 4 ℃ 12 h was taken to make blood natural coagulated and the supernatant was taken as the serum . Exosomes extracted by Ultracentrifugation , ExoQuick and Total Exosome Isolation Reagent ( TEI ) kits from serum were assessed by transmission electron microscopy , nanoparticle tracking analysis and protein analysis to identify morphology , protein expression , concentration and size distribution of particles .ExoRNA extracted by Trizol-LS, SeraMir, Total Exosome RNA Isolation ( TER) and exoRNeasy were assessed by Bioanalyzer 2100 and qPCR to ascertain RNA distribution and miRNA expression.Results For exosomes isolation , two commercial kits ( ExoQuick and TEI ) showed higher exosomes recovery and protein concentration than ultracentrifugation , but exosome isolated by ultracentrifugation expressed abundant protein makers mostly .For exo-RNA isolation, the distribution of RNA and expression of miRNAinfluenced by three methods , but there was no effect on the relative expression trend of miRNA.ExoRNeasy resulted in the highest yield and most narrow size distribution pattern of small RNA with higher level of miRNA expression .Results of TEI with TER kits showed no obvious bands of small RNA and moderate miRNA expression among methods .Ultra with Trizol-LS or ExoQuick with SeraMir showed low concentration measurable bands around 100 nt, with changeable miRNA profiling irregularly . Conclusions Extraction of exosomes using traditional ultracentrifugation method is applicable for proteomics research work .Extraction using ExoQuick and TEI kits is suitable for preparing exosomes from scarce samples such as serum.

OBJECTIVE:To study the pharmacokinetics and absolute bioavailability of oleanolic acid and its derivative di-hydrooleanolic acid in rats in vivo. METHODS:12 SD rats were randomly divided into oleanolic acid administration group and di-hydrooleanolic acid administration group,6 in each group. All rats were intragastrically given related medicine(50 mg/kg),then in-travenously injected related medicine(2 mg/kg)in tail vein after 1 week. Sample blood 0.3 mL was taken from tail vein before ad-ministration and after administration(0.1,0.25,0.5,0.75,1,1.5,2,3,5,7,9,12 h for ig;0.05,0.15,0.25,0.5,0.75,1, 2,3,5,7,9,12 h for iv). UPLC-QTOF was conducted to determine the plasma concentration,and DAS 2.0 pharmacokinetic software was used to calculate pharmacokinetic parameters and absolute bioavailability. RESULTS:After ig and iv oleanolic acid, the AUC0-∞ were(232.10±7.17),(1203.99±19.65)ng·h/mL,t1/2 were(1.75±0.10),(1.41±0.04)h respectively;after ig,cmax was (121.3 ± 18.92) ng/mL,tmax was (0.54 ± 0.10) h,absolute bioavailability was 0.77%. After ig and iv dihydrooleanolic acid, AUC0-∞ were(382.03±23.73),(386.14±10.65)ng·h/mL,t1/2 were(2.47±0.45),(1.44±0.03)h;after ig,cmax was(124.52± 12.28)ng/mL,tmax was(0.63±0.14)h,absolute bioavailability was 3.96%. CONCLUSIONS:The absolute bioavailability of di-hydrooleanolic acid is significantly higher than oleanolic acid in rats.

Objective To investigate the sex differences of the myelinated nerve fibers in the hippocampal formation of mid-aged standard and enriched rats. Methods Twenty 14-month Sprague-Dawley (SD) rats, 10 female and 10 male rats, were randomly divided into enriched group and standard group. Enriched rats were reared in enriched environment and standard rats were reared in standard environment for 4 months. Then, the total volume of the hippocampal formation and the myelinated nerve fibers in the hippocampus were quantitatively estimated by transmission electron microscopy and stereological methods. Results The total volume and mean diameter of the myelinated nerve fibers in the hippocampal formation of standard female rats were significantly bigger than those of standard male rats (P0.05). The mean diameter of the myelinated nerve fibers in the hippocampal formation of enriched female rats was significantly smaller than that of enriched male rats (P0.05). Conclusion The sex differences of the myelinated nerve fibers in the hippocampal formation of mid-aged standard and enriched rats are significantly different, which indicates the response to the enriched environment in the mid-aged male and female rats is different.

To develop a new small molecular probe for discovering an antitumor lead compound from the replacement of carboxylic group of two molecular antibacterial fluoroquinolones with a heterocyclic ring, a series of the C3/C3 bis-fluoroquinolones tethered with an 1, 3, 4-oxadiazole ring were synthesized as their respective HCl salts, and their structures were characterized by elemental analysis and spectral data. The in vitro antitumor activity against L1210, CHO and HL60 cell lines was also evaluated via the respective IC50 values by methylthiazole trazolium (MTT) assay.