MiR-21 promotes fracture healing by activating the PI3K/Akt signaling pathway.

OBJECTIVE: This study aims to elucidate the potential mechanism of micro ribonucleic acid (miR)-21 in promoting fracture healing. MATERIALS AND METHODS: 30 male Sprague-Dawley rats were randomly divided into group A (phosphate-buffered saline, PBS, n=10), group B (AntagomiR-21, n=10) and group C (AntagomiR-NC, n=10) according to the different treatments. The femoral fracture operation was performed in every rat, which was pathologically diagnosed via X-ray. After the successful modeling, 50 µL (2 nmoL) PBS, 50 µL AntagomiR-21 or 50 µL AntagomiR-NC was intraperitoneally injected into rats in group A, B or C, respectively. The above agents were injected once a week for 6 weeks. At 6 weeks, 3 rats were executed in each group, and the tissue at the fracture end was observed via hematoxylin-eosin (HE) staining. The fracture healing of rats was evaluated via imaging at 1 and 7 weeks. At the same time, the expression of miR-21 in the three groups was detected via Reverse Transcription-Polymerase Chain Reaction (RT-PCR), and the expression of phosphatidylinositol 3-hydroxy kinase (PI3K) and phosphorylated-serine/threonine-protein kinase B (p-Akt) in the three groups was detected via Western blotting. RESULTS: According to the histological staining, the bone repair at the fracture end of rats in group B was not significant with fracture and poor continuity compared with those in group A and group C. The imaging evaluation revealed that in group B, the callus tissues were significantly reduced, the fracture line had undesirable healing. There were no displacement and loosening of internal fixation compared with group A and group C. Besides, RT-PCR showed that the miR-21 expression declined markedly in group B compared with that in group A and group C, and the differences were statistically significant (p<0.05). Western blotting manifested that the protein levels of PI3K and p-Akt also declined in group B compared with those in group A and group C, and there were statistically significant differences (p<0.05). CONCLUSIONS: MiR-21 promotes the fracture healing in fractured rats by activating the PI3K/Akt signaling pathway.

[Short-term effect of Isobar dynamic stabilization system fixation combined with lumbar discectomy in patients with lumbar disc herniation].

Objective: To investigate the short-term effect of Isobar dynamic stabilization system fixation combined with lumbar discectomy in patients with lumbar disc herniation. Methods: From June 2015 to June 2017, 62 patients with lumbar disc herniation treated in the First Affiliated Hospital of Zhengzhou University were divided into control group and observation group according to the therapy. The 31 patients in the control group were treated with simple excision of nucleus pulposus and the 31 patients in the observation group were treated with Isobar dynamic stabilization system fixation combined with lumbar discectomy. The score of Oswestry disability index (ODI) and Japanese Orthopedic Association (JOA) score of low back pain, lumbar and adjacent stage activity, inflammatory factor levels[C reaction protein (CRP), interleukin 6 (IL-6) and tumor necrosis factor α (TNF-α)] were compared between the two groups before and after the operation and 6 months after the operation. The data were compared between the two groups with t test. Results: After the operation and 6 months after, the ODI scores of the two groups decreased and the JOA scores increased significantly (25.5±3.0, 27.5±3.2 vs 15.3±2.2 and 18.6±2.3, 23.3±2.9 vs 15.3±2.0), the ODI scores of the observation group was significantly lower than those in the control group; the JOA scores were significantly higher than those in the control group (t=0.04-10.19, all P<0.05). The operative time, hospital stay, intraoperative blood lose and complications rate in the observation group were all significantly lower than those in the control group. The total activity of lumbar vertebrae in the observation group was significantly higher than that in the control group after operation and 6 months after (t=37.67, 36.60, both P<0.05); the activity of adjacent segments in the observation group was significantly lower than that of the control group (t=9.28, 3.79, both P<0.05); the Pfirrmann grade was significantly lower than that in the control group (t=3.11, 5.05, both P<0.05). The levels of CRP, IL-6 and TNF-α in the two groups were lower than those before operation, and those were also significantly lower in the observation than the corresponding indexes in the control group (t=0.52-10.99, all P<0.05). Conclusion: Isobar dynamic stable system fixation combined with lumbar intervertebral disc resection can effectively improve the lumbar function and lumbar activity in patients with lumbar intervertebral disc herniation, and reduce the level of inflammation and relapse.

Effect of methionine deficiency on duodenal and jejunal IgA+ B cell count and immunoglobulin level of broilers.

Dietary methionine (met) is reported to enhance antibody production and boost cell-mediated immunity in chickens. Methionine deficiency has been shown to affect the development of the lymphoid organs and the generation of antibodies in chickens. This study is designed to investigate the effects of met deficiency on IgA+ B cells and immunoglobulins (sIgA, IgA, IgG and IgM) for a 6 week period in the duodenum and jejunum of Cobb broiler chicken using immunohistochemistry and enzyme-linked immunosorbent assay (ELISA) techniques. The results of the study showed that the IgA+ B cell count reduced significantly in the met deficiency group compared to the control group (P<0.05 or P<0.01). The contents of sIgA, IgA, IgG and IgM in the met deficiency group were significantly decreased (P<0.05 or P<0.01), especially at 28 and 42 days of age. It can be concluded that met deficiency exerts significant effects on the humoral immune system of intestinal mucosa. This study has provided valuable experimental insight which could be useful for future studies on the function of met in the intestine of humans and other animals.

Oxidative stress, apoptosis and abnormal expression of apoptotic protein and gene and cell cycle arrest in the cecal tonsil of broilers induces by dietary methionine deficiency.

Methionine (Met) is often called the first limiting amino acid in birds. Broilers have high requirement for Met, so they are at high risk of Met deficiency. The aims of the present research is to study the effects of Met deficiency on the histomorphological changes, antioxidant functions, apoptosis and cell cycle in the cecum tonsil of broilers. A batch of 120 one-day-old Cobb broilers in total are divided into two groups and fed on a Met deficiency diet or a control diet for six weeks, followed by analysis using the methods of experimental pathology, biochemical method, immunohistochemical method, ELISA, FCM, Tunel assay, and qRT-PCR. Results showed that the cecal tonsils were impaired, and the SOD, CAT and GSH-Px activity, the ability to suppression on the hydroxy radicals, and the content of GSH reduced in Met deficiency group comparing to the control group. In contrast, the MDA content is higher in Met deficiency group. As measured by immunohistochemical method, ELISA, Tunel, FCM and qRT-PCR, increased proportion of apoptotic cells, abnormal content or expression of apoptotic proteins, as well as cell cycle arrest were observed. In conclusion, dietary Met deficiency imposes a severe impact on the cecal tonsils, mainly in the forms of histological injury, cell cycle arrest, oxidative stress, and increased cellular apoptosis. The oxidative stress leads to cellular apoptosis and then induces the injury of the cecum tonsils. The local intestinal mucosal immune system would finally be injured by the oxidative stress and apoptosis in the intestine.

Care models in the management of haemophilia: a systematic review.

BACKGROUND: Haemophilia care is commonly provided via multidisciplinary specialized management. To date, there has been no systematic assessment of the impact of haemophilia care delivery models on patient-important outcomes. OBJECTIVE: To conduct a systematic review of published studies assessing the effects of the integrated care model for persons with haemophilia (PWH). SEARCH METHODS: We searched MEDLINE, EMBASE and CINAHL up to April 22, 2015, contacted experts in the field, and reviewed reference lists. SELECTION CRITERIA: Randomized and non-randomized studies of PWH or carriers, focusing mainly on the assessment of care models on delivery. DATA COLLECTION AND ANALYSIS: Two investigators independently screened title, abstract, and full text of retrieved articles for inclusion. Risk of bias and overall quality of evidence was assessed using Cochrane's ACROBAT-NRSI tool and GRADE respectively. Relative risks, mean differences, proportions, and means and their variability were calculated as appropriate. RESULTS: 27 non-randomized studies were included: eight comparative and 19 non-comparative studies. We found low- to very low-quality evidence that in comparison to other models of care, integrated care may reduce mortality, hospitalizations and emergency room visits, may lead to fewer missed days of school and work, and may increase knowledge seeking. CONCLUSION: Our comprehensive review found low- to very low-quality evidence from a limited number of non-randomized studies assessing the impact of haemophilia care models on some patient-important outcomes. While the available evidence suggests that adoption of the integrated care model may provide benefit to PWH, further high-quality research in the field is needed.

Clinical significance of quantitative monitoring and mutational analysis of BCR-ABL1 transcript in Philadelphia chromosome positive B lymphoblastic leukemia.

Quantitative detection of BCR-ABL1 transcript is essential in monitoring residual disease of Philadelphia chromosome positive B lymphoblastic leukemia (Ph+ B-LL). We studied the kinetics of BCR-ABL1 transcript in 41 Ph+ B-LL patients in correlation with their clinical outcome. A total of 23 patients achieved complete molecular remission at 6 months post-treatment. This was associated with a lower relapse risk and better overall survival. Likewise, sustainable complete molecular remission in 27 patients was associated with superior clinical outcome. Sporadic low level BCR-ABL1 was detected in 12 of 27 patients who had attained complete molecular remission. The relapse rate was significantly higher in non-transplant patients with persistent positive BCR-ABL1 than patients transplanted when BCR-ABL1 was detectable. All eight patients harboring ABL1 kinase domain mutations died of disease or were transferred to hospice care. We concluded that monitoring the level of BCR-ABL1 transcript after hematologic remission has predictive value to the long-term outcome.

Performance characteristics of the MultiCode-RTx hepatitis C virus load test targeting the 3' untranslated region.

Viral load testing for hepatitis C virus (HCV) RNA has become a key parameter in the diagnosis of infection and treatment monitoring. This study evaluated the performance characteristics of the MultiCode-RTx HCV assay (MultiCode; EraGen Biosciences, Inc., Madison, WI), a real-time PCR test targeting the 3' untranslated region (UTR) of the HCV genome, compared to the TaqMan HCV load ASR assay (TaqMan; Roche Diagnostics, Indianapolis, IN) that targets the 5' UTR. For plasma specimens, the MultiCode assay had a limit of detection of 2.3 log10 IU/ml and a linear range of at least 6.7 log10 IU/ml. Comparison of plasma viral loads obtained by the MultiCode and TaqMan tests showed that they were in very close agreement (mean difference, -0.1 log10 IU/ml). No genotype bias was observed for genotypes 1, 2, and 3. When the MultiCode assay was evaluated with the MagNA Pure and easyMAG extraction methods, the viral loads for the easyMAG extraction were consistently higher for all specimens tested (mean difference, 0.45 log10 IU/ml). Aside from the limit of detection, the performance characteristics of the MultiCode assay were similar to the TaqMan assay for the clinical application of HCV load testing.

Influence of maturation on constrictive response to stimulation of C-fiber afferents in isolated guinea pig airways.

We investigated whether the airway constrictive response to stimulation of bronchopulmonary C-fiber afferents is altered during the maturation process. Isometric tension was measured in airway rings isolated from three tracheobronchial locations (intrathoracic trachea and main and hilar bronchi) and compared in mature [M, 407 +/- 10 (SE) g body wt, n = 36] and immature (IM, 161 +/- 5 g body wt, n = 35) guinea pigs. Our results showed no difference in the ACh (10(-5) M)- or KCl (40 mM)-induced contraction between M and IM groups, regardless of the airway location. In sharp contrast, the concentration-response curves of 10(-8)-10(-6) M capsaicin were distinctly lower in IM hilar bronchi; for example, response to the same concentration of capsaicin (10(-6) M) was 89.2 +/- 15.3% of the response to 10(-5) M ACh in IM and 284.7 +/- 43.2% in M animals. Similar, but smaller, differences in the bronchoconstrictive response to capsaicin between IM and M groups were also observed in the trachea and main bronchus. Electrical field stimulation induced airway constriction in all three locations in M and IM groups. However, after administration of 10(-6) M atropine and 10(-6) M propranolol, electrical field stimulation-induced contraction was significantly smaller in the hilar bronchus of IM than M animals, and this difference was not prevented by pretreatment with 5 x 10(-5) M indomethacin. Although radioimmunoassay showed no difference in the tissue content of substance P between M and IM airways, the constrictive responses to exogenous substance P and neurokinin A were markedly greater in M airways at all three locations. In conclusion, the constriction of isolated airways evoked by C-fiber stimulation was significantly weaker in the IM guinea pigs, probably because of a less potent effect of tachykinins on the airway smooth muscle.