The continuing discoveries of novel classes of RNA modifications in various organisms have raised the need for improving sensitive, convenient, and reliable methods for quantifying RNA modifications. In particular, a subset of small RNAs, including microRNAs (miRNAs) and Piwi-interacting RNAs (piRNAs), are modified at their 3'-terminal nucleotides via 2'-O-methylation. However, quantifying the levels of these small RNAs is difficult because 2'-O-methylation at the RNA 3'-terminus inhibits the activity of polyadenylate polymerase and T4 RNA ligase. These two enzymes are indispensable for RNA labeling or ligation in conventional miRNA quantification assays. In this study, we profiled 3'-terminal 2'-O-methyl plant miRNAs in the livers of rice-fed mice by oxidative deep sequencing and detected increasing amounts of plant miRNAs with prolonged oxidation treatment. We further compared the efficiency of stem-loop and poly(A)-tailed RT-qPCR in quantifying plant miRNAs in animal tissues and identified stem-loop RT-qPCR as the only suitable approach. Likewise, stem-loop RT-qPCR was superior to poly(A)-tailed RT-qPCR in quantifying 3'-terminal 2'-O-methyl piRNAs in human seminal plasma. In summary, this study established a standard procedure for quantifying the levels of 3'-terminal 2'-O-methyl miRNAs in plants and piRNAs. Accurate measurement of the 3'-terminal 2'-O-methylation of small RNAs has profound implications for understanding their pathophysiologic roles in biological systems.
Animals ; High-Throughput Nucleotide Sequencing ; Humans ; Methylation ; Mice ; MicroRNAs/genetics* ; Oxidative Stress ; RNA, Small Interfering/metabolism* ; Real-Time Polymerase Chain Reaction

Objective To evaluate the efficacy of WeChat platform conbined with problem-based learning (PBL) teaching method when applied to the standardized training for residents in a department of anesthesiology.Methods Ninety-six residents who were undergoing the standardized training in the Department of Anesthesiology of our hospital were divided into 3 groups (n =32 each) using a random number table:lecture-based learning teaching method group,PBL teaching method group and WeChat platform plus PBL teaching method group.The examination scores after the end of teaching,satisfaction with teaching and scores of interest in anesthesiology were recorded.Results The total scores of examination,scores of comprehension memory problems,clinical application problems and comprehensive analysis problems,rate of satisfactory teaching and scores of interest in anesthesiology were significantly higher in WeChat platform plus PBL teaching method group than in the other two groups (P＜0.05).Conclusion WeChat platform combined with PBL teaching method can improve the teaching quality of the standardized training for residents in a department of anesthesiology and win praise from residents,and it is worth promoting the application.

MicroRNAs (miRNAs) are small, non-coding RNAs that function as post-transcriptional regulators of gene expression. The deregulated expression of miRNAs is associated with a variety of diseases, including breast cancer. In the present study, we found that miR-495 was markedly up-regulated in clinical breast cancer samples by quantitative real time-PCR (qRT-PCR). Junctional adhesion molecule A (JAM-A) was predicted to be a potential target of miR-495 by bioinformatics analysis and was subsequently verified by luciferase assay and Western blotting. JAM-A was found to be negatively correlated with the migration of breast cancer cells through loss-of-function and gain-of-function assays, and the inhibition of JAM-A by miR-495 promoted the migration of MCF-7 and MDA-MB-231 cells. Furthermore, overexpression of JAM-A could restore miR-495-induced breast cancer cell migration. Taken together, our findings suggest that miR-495 could facilitate breast cancer progression through the repression of JAM-A, making this miRNA a potential therapeutic target.
3' Untranslated Regions ; genetics ; Adult ; Aged ; Blotting, Western ; Breast Neoplasms ; genetics ; metabolism ; pathology ; Cell Adhesion Molecules ; genetics ; metabolism ; Cell Line, Tumor ; Cell Movement ; genetics ; Female ; Gene Expression Regulation, Neoplastic ; Gene Knockdown Techniques ; Humans ; MCF-7 Cells ; MicroRNAs ; genetics ; Middle Aged ; RNA Interference ; Receptors, Cell Surface ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction

Humans ; Interferon-gamma ; analysis ; Lymphohistiocytosis, Hemophagocytic ; diagnosis ; microbiology

In order to analyze the genotype of RhD-negative blood donors with immune antibodies in Harbin, the voluntary blood donors from 1 April 2008 to 30 september 2011 were detected serologically to determine the RhD-negative donors. The blood donors confirmed to be RhD negative were detected to screen the immune antibodies, the samples with immune antibodies were analyzed by PCR-SSP and DNA sequencing to detect RhD genotype. The results showed that the 12 cases of the immune antibodies (0.95%) were screened out from 1265 cases of RhD-negative donors, among which 9 cases showed anti-D-antibody, 3 cases showed anti-(D+C) antibody; 10 cases were RhD-negative, 2 cases were RHD 711D(el)C. It is concluded that RhD negative and RHD 711D(el)C are easy to be immunized to produce the immune antibodies; RhD-negative population, especially women should be highly aware of avoiding mis-transfusion of RhD-positive blood, and also avoiding multiple pregnancies resulting in newborn's hemolytic disease.
Base Sequence ; Blood Donors ; Exons ; Genotype ; Humans ; Isoantibodies ; Phenotype ; Rh-Hr Blood-Group System ; genetics ; immunology ; Rho(D) Immune Globulin

Objective To assess the association of synovial cyclic citrullinated peptide(CCP)expression with T helper 17(Th17) cells/Regulatory T cells (Treg) imbalance,the histological and clinical features of synovitis in rheumatoid arthritis (RA).Methods CCP expression in synovial specimens from 39 patients with RA and 35 controls was detected by immunohistochemistry assay(IH) using 6×His tagged anti-CCP single chain fragment V (ScFv) antibodies,which were generated by pHEN2 phagemid recombinant antibodies display system.The frequencies of Th17/Treg cells in the peripheral blood mononuclear cells (PBMCs) were determined by flow cytometry (FCM).Th17/Treg cells associated cytokines were analyzed by enzyme-linked immunosorbent assay (ELISA).The histological scores and clinical features of synovitis were included in the study.Chi-square test and ANOVA were used for statistical analysis.Results ① The prevalences of synovial C CP expression were significantly different between RA group and the control(76.9％ and 11.4％ respectively,X2=31.9,P＜0.01).② The frequencies of Th17 cells,Th17/Treg ratio,Th17 cells associated cytokines as IL-6,IL-17a,IL-23,TNF-α,and the Treg cells associated cytokines TGF-31,the serum and synovial fluid anti-CCP antibodies in the RA patients with synovial CCP positive expression were significantly higher in RA patients with CCP positive than those with CCP negative.Disease activity score DAS28 index and histological features quantified variations of the synovial biopsy specimens (synoviocyte hyperplasia,focal aggregates of lymphocytes,and diffuse infiltrat(e)s of lymphocytes) in RA were higher in synovial CCP positive expression patients than in the negative.Conclusion Synovial CCP expression is strongly associated with the Th17/Treg imbalance and synovitis,which may play a crucial role in the pathogenesis of RA.

OBJECTIVETo comparing two kinds habitat adaptive division of Chinese material medica with different models.

METHODThe habitat adaptive divisions of A. lancea according essential oil accumulation with two kinds pattern, model pattern and template pattern were carrid and compared.

RESULTTwo habitat adaptive divisions of A. lancea maps according essential oil accumulation were gotten.

CONCLUSIONBoth model pattern and template pattern were efficient on habitat adaptive division of Chinese material medica, but they shoud bu used with different processesd and based different background [corrected]

Atractylodes ; metabolism ; Ecosystem ; Materia Medica ; Medicine, Chinese Traditional ; methods ; Oils, Volatile ; metabolism

Objective To study hepatic blood flow exclusion for the resection of liver tumors involving hepatic hilar region. Methods The clinical data of 16 cases of liver tumors involving hepatic hilar region from January 2005 to March 2008 were retrospectively analyzed. Liver tumors were resected by the technique combining hepatic portal control ( Pringle's maneuver) and normothermie total hepatic vascular exclusion (NHVE). The relation of liver tumors to major vessels, episodes and durations of hepatic blood flow exclusion, intraoperative blood loss and blood transfusion, postoperative complications were analyzed. Results The technique combining Pringle's maneuver and NHVE was used in 16 cases. The mean episodes and durations of Pringle's maneuver were (3.8±1.6) min and (46.6±28.8) min, respectively. The mean episodes and durations of NHVE were (1.6±0.4) min and (23.5±8.2) min, respectively. The mean amount of intraoperativ blood loss was (1250±320) ml, blood transfusion (860±245) ml. Major hepatic vessel injuried were repaired intraoperatively including inferior vena cave in 4 cases, main hepatic veins in 2 cases and portal veins in 2 cases. The serum alanine transaminase(ALT) and bilirubin raised in different degrees after operation, and recovered gradually to normal level. There was no postoperative mortality and serious postoperative complications. Conclusions Alternative use of hepatic blood flow exclusion combining Pringle maneuver and NHVE reduces the time of total hepatic vascular exclusion, improves safety for resection of liver tumors involving hepatic hilar region.

OBJECTIVETo investigate the role of xenogenic (porcine) ADM as dermal substitute in scar treatment.

METHODSAfter scar excision, the wounds were covered with composite grafts of DR procine ADM and autologous thin split-thickness grafts in one stage or in two stages.

RESULTS22 out of 47 cases were treated in two-staged procedure. After the ADMs were applied to the wound, the autologous thin split-thickness grafts were implanted 7 days later. 25 cases were treated in one-staged procedure. The survival rates of composite grafts were (88.3 +/- 3.7)% for subcutaneous recipient bed and (89.7 +/- 3.4)% for deep fascia recipient bed in group with two-staged procedure, compared with (92.5 +/- 4.1)% and (93.2 +/- 5.2)%, respectively, in group with one-staged procedure. Early after grafts taken, the grafts had a pink colour and smooth surface. The patients were followed up for 90 days at most. The survived composite grafts were durable, elastic, smooth and soft with good function and appearance like normal skin. They could even be pinched up. The scar along the edge of the grafts was slightly hypertrophic.

CONCLUSIONSThe survival rate of composite graft is higher in patients with one-staged procedure. The elasticity and textural of the taken grafts are better on subcutaneous recipient bed than on deep fascia recipient bed, though the function has no difference. Xenogenic (porcine) ADM can be an optimal dermal substitute for wound coverage after scar excision.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Animals ; Child ; Child, Preschool ; Cicatrix ; surgery ; Dermis ; cytology ; transplantation ; Humans ; Male ; Middle Aged ; Skin, Artificial ; Swine ; Transplantation, Heterologous ; Young Adult

BACKGROUNDNitric oxide (NO) is an important mediator in the pathophysiology of many vascular diseases. However, the definite role of NO in human abdominal aortic aneurysm (AAA) formation is unclear. The aim of this study was to investigate production of NO and expression of inducible nitric oxide synthase (iNOS), and their possible role in AAA.

METHODSA total of 28 patients with AAA, 10 healthy controls, and 8 patients with arterial occlusive disease were enrolled into this study. Standard colorimetric assay was used to examine NO concentration in plasma from patients with AAA and normal controls, and in cultured smooth muscle cells (SMCs). Expression of iNOS in aortas and cultured SMCs were detected by immunochemistry. The correlation of iNOS expression with age of the patient, size of aneurysm, and degree of inflammation was also investigated by Cochran-Mantel-Haenszel chi2 test and Kendall' Tau correlation.

RESULTSExpression of iNOS increased significantly in the wall of aneurism in the patients with AAA compared to the healthy controls (P < 0.05) and the patients with occlusive arteries (P < 0.05). iNOS protein and media NOx (nitrite + nitrate) also increased in cultured SMCs from human AAA (n = 4, P < 0.05), while plasma NOx decreased in patients with AAA (n = 25) compared to the healthy controls (n = 20). There was a positive correlation between iNOS protein and degree of inflammation in aneurismal wall (Kendall coefficient = 0.5032, P = 0.0029).

CONCLUSIONSSMCs and inflammatory cells were main cellular sources of increased iNOS in AAA, and NO may play a part in pathogenesis in AAA through inflammation.

Adult ; Aged ; Aortic Aneurysm, Abdominal ; etiology ; Apoptosis ; Female ; Humans ; Male ; Middle Aged ; Muscle, Smooth, Vascular ; pathology ; Nitric Oxide ; physiology ; Nitric Oxide Synthase Type II ; analysis ; physiology