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Objective:To explore the effects of conditioned medium of human bone marrow mesenchymal stem cells (BMSCs) on the proliferation, adhesion and differentiation of immortalized human Müller cell line (MIO-M1).Methods:The differentiation was induced in the third-passage BMSCs with osteogenic, chondrogenic and adipogenic medium and identified by alizarin red, alcian blue and oil red O staining, respectively.The expression levels of mesenchymal stem cell markers CD73, CD90 and CD105 and hematopoietic cell markers CD34, CD45 and human leukocyte antigen-DR (HLA-DR) were assayed by flow cytometry.The expressions levels of Müller cell markers SOX9, glutamine synthetase (GS), vimentin and cellular retinaldehyde-binding protein (CRALBP), retinal stem cell markers SOX2, nestin and CHX10, and cell proliferation marker cyclin D3 (CCND3) in MIO-M1 cells were detected by immunofluorescence staining.The MIO-M1 cells were divided into standard medium group, 293T conditioned medium group, and BMSC conditioned medium group and were incubated in the medium according to grouping.The cellular area, circularity, elongation factor and perimeter were analyzed quantitatively.The cell cycle was detected by flow cytometry, and the cell proliferation was determined by neurospora experiment and 5-ethynyl-2'-deoxyuridine (EdU) staining.The expression of vascular cell adhesion molecule 1 (VCAM-1) at protein and mRNA levels in the culture supernatant was detected by enzyme linked immunosorbent assay (ELISA) and quantitative real-time PCR (qRT-PCR), respectively.The expression of retinal neuron markers protein kinase C (PKCα), Rhodopsin, microtubule-associated protein 2 (MAP2) and β-tubulin (Tuj1) was detected by immunofluorescence staining and qRT-PCR.Results:CD73, CD90, CD105 showed an enhanced expression, and CD34, CD45 and HLA-DR showed weakened expression in the BMSCs.The BMSCs differentiated into osteoblasts, chondrocytes and adipocytes.Expression of SOX9, GS, vimentin and CRALBP, SOX2, CHX10, nestin and CCND3 was found in the MIO-M1 cells.Compared with standard medium group and 293T conditioned medium group, MIO-M1 cells cultured in BMSC conditioned medium group changed into an elongated spindle-shaped or multipolar morphology with reduced cell area, increased elongation index and decreased circularity, showing statistically significant differences among them ( F=6.973, 12.370, 6.311; all at P<0.01). There were increased neurospheres formed by MIO-M1 cells in BMSC conditioned medium group compared with standard medium group and 293T conditioned medium group at different time points ( Fgroup=134.300, P<0.001; Ftime=82.910, P<0.001). Compared with the standard medium group and 293T conditioned medium group, the EdU-positive rate and proliferation index of MIO-M1 cells in BMSC conditioned medium group were significantly increased, with statistically significant differences ( F=6.973, 74.110; all at P<0.05); the VCAM-1 protein expression in cell supernatant and the relative expression level of VCAM-1 mRNA in BMSC conditioed medium group were significantly increased ( F=13.720, 7.896; all at P<0.05); the mRNA expression levels of PKCα, Rhodopsin, Tuj1 and MAP2 were higher in MIO-M1 cells of BMSC conditioned medium group under the condition of differentiation ( F=14.490, 5.424, 14.330, 7.405; all at P<0.05). Conclusions:BMSCs conditioned medium can change the morphology of MIO-M1 cells and promote their proliferation, adhesion and differentiation into retinal neurons.
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@#Reoperation due to degenerated bioprostheses is an important factor of high-risk thoracic surgeries. In 2020 ACC/AHA guideline, Valve in Valve (ViV) was recommended for high-risk patient instead of surgical mitral valve replacement. This report described a 77-year-old male patient with a failed mitral bioprosthetic valve, evaluated at high risk of surgery, received a transvenous, transseptal transcatheter mitral valve replacement (TMVR). Tracheal intubation was removed at CCU 3 h after surgery without discomfort such as polypnea. The patient was transferred out of the CCU and discharged on the 3rd day. Compared with transapical access, transvenous transseptal access was less invasive, with shorter duration in CCU and hospitalization.
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In previous studies, we found that truncated rotavirus VP4* (aa 26-476) could be expressed in soluble form in Escherichia coli and confer high protection against rotavirus in the mouse mode. In this study, we further improved the immunogenicity of VP4* by polymerization. The purified VP4* was polymerized through incubation at 37 ℃ for 24 h, and then the homogeneity of the particles was analyzed by HPLC, TEM and AUC, while the thermal stability and antigenicity was analyzed by DSC and ELISA, respectively. Finally, the immunogenicity and protective efficacy of the polymers analyzed by a mouse maternal antibody model. The results showed that VP4* aggregated into homogeneous polymers, with high thermostability and neutralizing antibody binding activity. In addition, VP4* polymers (endotoxin <20 EU/dose) stimulated higher neutralizing antibodies and confer higher protection against rotavirus-induced diarrhoea compared with the VP4* trimers when immunized with aluminium adjuvant. In summary, the study in VP4* polymers provides a new strategy for the development of recombinant rotavirus vaccines.
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Animals , Mice , Antibodies, Viral , Antigens, Viral , Capsid , Capsid Proteins , Polymerization , Rotavirus , Rotavirus InfectionsABSTRACT
Objective To evaluate the value of procalcitonin(PCT) and endotoxin(ET) in early diagnosis and prognosis in critically illness patients with sepsis.Methods A total of 104 cases with suspected sepsis were prospectively collected in this study from February 2015 to October 2016 in ICU.There were 31 cases of systemic inflammatory response syndrome (SIRS group),30 cases of sepsis 24 cases of severe sepsis and 19 cases of septic shock.Prognosis was observed in patients with sepsis discharged,57 cases were survival (survival group) while 16 cases were dead (death group).The level of PCT and ET and the clinical characteristics were recorded,The the ROC curve was applied to evaluate the discriminative power of variables.Results The levels of PCT,APACHE-Ⅱ score,ET of cases with sepsis,severe sepsis and septic shock were significantly higher than those in SIRS group(P0.05).ROC curve showed that AUC of PCT was 0.867,which was significantly higher than that for APACHE-Ⅱ score (AUC=0.762) (P>0.05).Conclusion Compared with APACHE-Ⅱ score and ET,PCT can help clinicians to early diagnose and treat cases with sepsis.
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Objective To observe the clinical effect of doxazosin combined with diclofenac sodium in the treatment of bladder spasm after transurethral bipolar plasmakinetic prostatectomy (TUPKP).Methods 200 BPH patients undergoing TUPKP were randomly divided into 4 groups according to single-blind randomized controlled trial design:group A,group B,group C,group D (n =50,each).Patients in group A were given diclofenac sodium 100 mg,q12h,in anus after surgery and doxazosin 4 mg/d orally after anesthetic awareness.Patients in group B were given diclofenac sodium in anus 100 mg,q 12h in anus after surgery.Patients in group C were given doxazosin 4 mg/d orally after anesthetic awareness.Patients in group D were not given diclofenac sodium and doxazosin.Drugs were withdrawn 3 days after surgery.Results In group A,the average time of bladder spasm was (0.47±0.18) time,(0.35±0.16) time,(0.30±0.20) time at the 1st,2 nd,3 rd day respectively; the duration of bladder spasm was (3.2±1.5) min,(2.1±1.3) min,(1.4±1.2)min at the 1st,2nd,3th day respectively; the time of bladder perfusion clearance was (1.5± 0.3)days and the time of urethral catheter removal was (4.0±0.5) days.There were significant differences in above observed values between group D and the other groups (all P＜0.05).Conclusions Doxazosin combined with diclofenac sodium therapy is effective in the treatment of bladder spasm after bipolar transurethral plasmakinetic prostatectomy.