LRRC25 plays a key role in all-trans retinoic acid-induced granulocytic differentiation as a novel potential leukocyte differentiation antigen

Leukocyte differentiation antigens (LDAs) play important roles in the immune system, by serving as surface markers and participating in multiple biological activities, such as recognizing pathogens, mediating membrane signals, interacting with other cells or systems, and regulating cell differentiation and activation. Data mining is a powerful tool used to identify novel LDAs from whole genome. LRRC25 (leucine rich repeat-containing 25) was predicted to have a role in the function of myeloid cells by a large-scale "omics" data analysis. Further experimental validation showed that LRRC25 is highly expressed in primary myeloid cells, such as granulocytes and monocytes, and lowly/intermediately expressed in B cells, but not in T cells and almost all NK cells. It was down-regulated in multiple acute myeloid leukemia (AML) cell lines and bone marrow cells of AML patients and up-regulated after all-trans retinoic acid (ATRA)-mediated granulocytic differentiation in AML cell lines and acute promyelocytic leukemia (APL; AML-M3, FAB classification) cells. Localization analysis showed that LRRC25 is a type I transmembrane molecule. Although ectopic LRRC25 did not promote spontaneous differentiation of NB4 cells, knockdown of LRRC25 by siRNA or shRNA and knockout of LRRC25 by the CRISPR-Cas9 system attenuated ATRA-induced terminal granulocytic differentiation, and restoration of LRRC25 in knockout cells could rescue ATRA-induced granulocytic differentiation. Therefore, LRRC25, a potential leukocyte differentiation antigen, is a key regulator of ATRA-induced granulocytic differentiation.

Relationship between the lumbar quantitative computed tomography values and contrast agent dispersion in osteoporotic thoracolumbar fractures / 中国组织工程研究

BACKGROUND: Percutaneous vertebroplasty (PVP) is usually used for osteoporotic thoracolumbar fractures,which has various advantages such as easy to operate, short operation time, less trauma, rapid recovery,analgesic effect and so on. But its application is restricted due to nerve compression symptoms and pulmonary embolism caused by bone cement leakage. Thereafter, how to reduce the leakage of bone cement is an issue of concern.OBJECTIVE: To investigate the relationship between the lumbar quantitative computed tomography (QCT) values and contrast agent dispersion in osteoporotic thoracolumbar fractures. METHODS: Sixty cases of osteoporotic thoracolumbar fractures undergoing PVP were enrolled, and received QCT examination before surgery, and contrast agent was injected intraoperatively. X-ray examination was conducted to detect the bone mineral density, contrast agent dispersion and leakage of bone cement, and the relationship between the lumbar QCT values and contrast agent dispersion as well as leakage of bone cement.RESULTS AND CONCLUSION: (1) There were 110 vertebral fractures, and 74 vertebrae with contrast agent diffusing more than vertebral midline, accounting for 67.3%. There was significant difference in the contrast agent dispersion among groups (P 0.05). (3) These results suggest that contrast agent dispersion in osteoporotic thoracolumbar fractures has a certain relationship with the lumbar QCT values, and lumbar QCT values with more contrast agent dispersion, but the lumbar QCT values have no correlation with bone cement leakage. Therefore, choosing a appropriate approach based on the QCT values and contrast agent dispersion can reduce leakage and improve the safety of PVP.

Serum protein fingerprint pattern model for diagnosing blood stasis syndrome of lumbar intervertebral disc protrusion / 中国组织工程研究

BACKGROUND:The correlation between blood stasis syndrome and non-blood stasis syndrome of lumbar intervertebral disc protrusion remains unclear. OBJECTIVE:To construct serum protein pattern model for diagnosing blood stasis syndrome of lumbar intervertebral disc protrusion. METHODS:A total of 180 cases were included in this study and divided into treatment group (120 patients with lumbar intervertebral disc protrusion) and control group (60 healthy cases from physical examination). Furthermore treatment group was equal y assigned into blood stasis syndrome subgroup and non-blood stasis syndrome subgroup, with 60 cases in each subgroup. The involved cases were wel matched in nations, genders and ages. Serum samples of peripheral blood from the 180 cases were col ected. Surface-enhanced laser desorption/inionation time of flight mass spectrometry and ProteinChip technology were employed to detect and plot protein mass spectrum. The protein peak values were identified using Biomarker Wizard software. Then serum diagnosis model of blood stasis syndrome of lumbar intervertebral disc protrusion was established. The obtained models were verified through double blind method. The differential proteins were searched by ExPASy data. RESULTS AND CONCLUSION:We detected that peak values of eleven proteins had statistical significance (P<0.05) from the involved 180 cases. Among them, two proteins were highly expressed while the other nine proteins were lowly expressed. Serum protein pattern model for diagnosing blood stasis syndrome of lumbar intervertebral disc protrusion was established through Biomarker Patterns software, and the sensibility was 86.667%, the specificity was 94.167%, the positive predictive value was 88.136%. There are a variety of abnormal y expressed proteins in the serum of the patients with blood stasis syndrome of lumbar intervertebral disc protrusion. The serum protein pattern model involved eleven different proteins can be used to diagnose blood stasis syndrome of lumbar intervertebral disc protrusion.

Preparation and characterization of the antibodies against human myofibrillogenesis regulator 1 and application in neonatal rat cardiomyocytes / 中国病理生理杂志

AIM: To prepare and purify the polyclonal antibodies against human myofibrillogenesis regulator 1 (hMR-1), then to characterize the purity, titer, specificity and the availability.METHODS: Two polypeptides named peptide 1 and 2 were synthesized based on the bioinformatics analysis of the sequence of hMR-1 by using software TMHMM and DNAStar, then coupled with keyhole limpet hemocyanin (KLH) for immunization. These peptides for immunization were mixed and injected into New Zealand rabbits to prepare antibodies specifically against hMR-1. ELISA assay was used to detect the titers of the antibodies. After purification by immunoaffinity chromatography, antibodies were identified by Western blotting and immunocytofluorescent assays. Applications of the antibodies on neonatal rat cardiomyocytes were also employed.RESULTS: (1)The titers of antibodies were 1:10~5. In WB assay, a specific 17kD band was detected, corresponding to the predicted molecular weight of hMR-1; the positive fluorescent signals were distinct. (2)On the neonatal rat cardiomyocytes model, we observed a peri-nucleus location. The fluorescent signal of hMR-1 overexpression group was much stronger than that in vector control and normal control groups.CONCLUSION: All these results indicate that the antibodies obtained from poly peptides mixture immunization have either human original or rat original antigens. The antibody is available for using in Western blotting or immunofluorescent assays.

Effect of Simvastatin-Polylactic Acid Compound on Critical Calvarial Defect / 中国微创外科杂志

Objective To investigate the effect of simvastatin-polylactic acid compound on critical calvarial defects in rats.Methods Twenty male SD rats(150 g?10 g),were used to establish critical cranial defect(10 mm in diameter)model.The animals were randomly divided into control and experiment groups(10 in each).In the control group,40 mg of polylactic acid were implanted into the defect area;whereas in the experiment group,simvastatin-polylactic acid compound were used(20 mg simvastatin and 40 mg polylactic acid).Four and eight weeks after the implantation,the defect area of the rats was observed by X-ray and toluidine blue staining.Results Eight weeks after the operation,X-ray examination showed high-density regions in the defect area in the experiment group,while low-density regions in the control group.The radiopacity of cranial defects were 27.33%?2.54% in the control group,and 74.63%?2.42% in the experimental group(n=5,t=-30.148,P=0.000).Toluidine blue staining showed a few new bone tissues at 4 weeks and fully filled bone defect at 8 weeks in the experiment group.Meanwhile,in the control group,only a small quantity of new bone tissue could be seen on the edge of the cranial defects.Conclusion Locally implanted simvastain-polylactic acid compound is a promising method for the treatment of bone defect owing to its high osteogenic ability.

Effect of Simvastatin on Mobilization and Migration of Endothelial Progenitor Cells / 中国微创外科杂志

Objective To investigate the effect of simvastatin on the mobilization and migration of endothelial progenitor cells(EPCs).Methods EPCs were harvested from the bone marrows of two rabbits,cultured with M199,and identified by immunohistochemistry.The identified EPCs were then treated with simvastatin with different concentrations(0,0.01,0.1,1.0 ?mol/L),and their migration induced by simvastatin was determined with Transwell chamber assay.Six rabbits models of cranial bone defect were established and divided into control and experiment groups(3 in each).In order to elicit the effects of simvastatin on mobilization of EPCs,simvastatin was embedded in polylactic acid compound,and implanted into the cranial bone defect area in the experiment group.Meanwhile,polylactic acid was implanted in the control animals.After 10 days,the expression rate of CD34+/CD133+ EPCs in the rabbit peripheral blood was counted by flow cytometry to determine the motivating effect of simvastatin.Results In Transwell experiment,16 hours after adding simvastatin(0,0.01,0.1 or 1.0 ?mol/L),the cell migration ability was obviously increased showing a dose-dependent trend(OD value:0.077?0.014 in control group and 0.075?0.013 in 0.01 ?mol/L group vs 0.097?0.011 in 0.1 ?mol/L group and 0.099?0.019 in 1.0 ?mol/L group,P

CMTM1-v17, a new potential corepressor of androgen receptor / 北京大学学报(医学版)

Objective: To analyze the expression of CMTM1-v17 in normal prostate tissue and prostate carcinoma originated cell lines, and study its impact on the transactivation of androgen receptor and the possible mechanism. Methods: The expression of CMTM1-v17 in normal prostate tissue was analyzed with immunohistochemistry method. In immounocytochemistry was used to analyze the expression of CMTM1-v17 in prostate carcinoma originated cell lines. Luciferase assay was used to study the impact of CMTM1-v17 on the transactivation of AR and its mechanism. Results: The results of immunohistochemistry showed that CMTM1-v17 was highly expressed in prostate. In prostate cancer originated cell lines, CMTM1-v17 could also be detected in prostate cancer originated cell lines PC3, Du145 and LNCaP. And the results of luciferase implied that the relative luciferase activity of the PC3 cells transfected with 1 ?g and 2 ?g pCDI-CMTM1-v17 plasmids separately were 70.8 and 34.7, compared with the control set as 100. When trichostatin A, the inhibitor for histone deacetylase, was used, the repression of androgen receptor could be recovered with trichostatin A treatment,for the relative luciferase activity of the PC3 cells transfected with 1 ?g and 2 ?g pCDI-CMTM1-v17 plasmids and treated with 100 nmol/L trichostatin A rebound to 90.9 and 86.4. Conclusion: CMTM1-v17 is highly expressed in both normal prostate and prostate carcinoma originated cell lines. It may recruit histone deacetylas to inhibit the function of androgen receptor.

Nuclear translocation of apoptosis-related protein TFAR19 in TF-1 cells undergoing apoptosis / 北京大学学报(医学版)

Objective: To study the expression and localization of apoptosis-related protein TFAR19 in TF-1 cells undergoing apoptosis. Methods: Using monoclonal antibody against TFAR19, the expression level and cell localization of TFAR19 were examined by fluorescence microscope, confocal laser scan microscope(CLSM) and flow cytometry. Simultaneously, we also analyzed the relationship of TFAR19 protein with phosphatidylserine (PS) externalization and cell nuclear DNA fragmentation. Results: The level of TFAR19 proteins expressed in TF-1 cells treated with GM-CSF withdrawal was significantly increased compared with normal TF-1 cells, then translocated rapidly from cytoplasm to the nucleus of cells. Appearance of TFAR19 in the nucleus of apoptotic cells preceded the detection of PS externalization and DNA fragmentation. Conclusion: Nuclear translocation of TFAR19 protein is one of the earliest events of cell apoptotic process. These data provided a new clue to further approach to the biological function of TFAR19 and study of cell apoptosis.