BACKGROUND:Autologous bone marrow mesenchymal stem cel transplantation for the treatment of cardiovascular disease has become one of the hotspots, but it is unclear whether the proliferation and directional differentiation of autologous bone marrow mesenchymal stem cels varies changes with age. OBJECTIVE: To explore the proliferation and differentiation changes of rat bone mesenchymal stem cels in different ages. METHODS:The bone marrow mesenchymal stem cels from Sprague-Dawley rats in different age groups were purified and cultured, and then examined by flow cytometry in terms of cel cycle. Meanwhile, neonatal rat cardiomyocytes were co-cultured with bone marrow mesenchymal stem cels. The morphologic changes of bone marrow mesenchymal stem cels and the protein expression of troponin T were detected with immunofluorescence technique. RESULTS AND CONCLUSION: The percentage of bone marrow mesenchymal stem cels in G0/G1 phase was increased with age; while the percentage of expression of troponin T proteins-positive bone marrow mesenchymal stem cels were decreased with age. These findings indicate that the proliferation and differentiation abilities of rat bone marrow mesenchymal stem cels descend with age.

AIM:To analyze the correlation between serum angiotensin-converting enzyme 2 (ACE2) levels and different stages of coronary heart disease (CHD), and to explore the change of serum ACE2 level during the development of CHD.METHODS:The control group included 85 non-CHD samples, and 174 CHD samples were divided into light stenosis (ls-CHD, stenosis degree <50%) group, moderate stenosis (ms-CHD, stenosis degree 50%~75%) group and severe stenosis (ss-CHD, stenosis degree ≥75%) group.The ACE2 level in each serum sample was detected by ELISA.The relationship between the ACE2 level and the development of coronary heart disease was explored by statistical analysis of serum ACE2 levels in different stages of CHD.RESULTS:The serum ACE2 levels in ls-CHD group, ms-CHD group and ss-CHD group were all higher than that in control group.The more severe the coronary artery stenosis existed, the higher the ACE2 level was observed.The serum ACE2 level in the males was higher than that in the females.In a single sex, the serum ACE2 levels in ls-CHD group, ms-CHD group and ss-CHD group were higher than that in control group with significant differences.Regression analysis found that sex, diabetes and CHD were associated with the serum ACE2 levels.Among them, sex and CHD were the independent factors to affect serum ACE2 levels.CONCLUSION:The serum ACE2 level of males was higher than that of females.Compared with the non-CHD samples, the serum ACE2 level of CHD patients was higher than that of the non-CHD samples.During the development of coronary heart disease, the serum ACE2 level increased constantly.

To get the mature peptide of human-derived neurotrophin-6 (NT-6), NT-6 gene encoding mature peptide was amplified by PCR, using the NT-6 cDNA that had been cloned as templet. The gene encoding mature peptide of NT-6 gene was cloned into pGEX1-lambdaT plasmid to construct the fusion expression vector. Expression of fusion protein in Escherichia coli was defected after induction by isopropyl beta-D-thiogalactoside(IPTG). The mature peptide of NT-6 was collected with GST fusion protein purifying kit. It was shown that a fragment of 460bp was gained by PCR. With the techniques of double-cleave and electrophoresis, the recombinant vector was identified as pGEX1-NT-6. The recombinant vector pGEX1-NT-6 transformed Escherichia coli expressed fusion protein of 41KD after induction by IPTG. Cleaved by thrombin, the mature peptide of NT-6 was obtained; its molecular weight was about 15KD. The cloning and expression of human-derived NT-6 gene encoding mature protein has provided a basis for further studies on the function and clinical application of NT-6.
Escherichia coli ; genetics ; metabolism ; Genetic Vectors ; genetics ; Humans ; Isopropyl Thiogalactoside ; pharmacology ; Nerve Growth Factors ; biosynthesis ; genetics ; Peptides ; genetics ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; isolation & purification

Objective To compared the macular blood flow parameters among myopic choroidal neovascularization (mCNV), high myopia (HM) and normal subjects.Methods Retrospective study. Forty patients (40 eyes) diagnosed as mCNV (mCNV group) in the Eye Hospital of Wenzhou Medical University at Hangzhou from June 2016 to November 2018, age-matched, sex-matched and diopter-matched 40 HM patients (40 eyes, HM group), age-matched and sex-matched 40 healthy persons (40 eyes, normal group) were enrolled in this study. Retina superficial and deep vessel density, the area of foveal avascular zone (FAZ), a-circularity index (AI) and vessel density around the 300 μm width of the FAZ region (FD) on macular 3 mm×3 mm scan on OCTA of 3 groups were compared.Results There were significant differences in the average retina superficial, deep vessel density, the area of FAZ, AI and FD among 3 groups (F=24.82, 9.18, 3.58, 7.68, 14.15;P<0.05). The average retina superficial and deep vessel density and FD in mCNV group were lower than those in HM group (P<0.05). The average retina superficial and deep vessel density and FD in HM group were lower than those in control group (P<0.05). The average area of FAZ in mCNV group was smaller than that in control group (P<0.05). The average AI in mCNV group was less than that in the other 2 groups (P<0.05). Conclusions The retina superficial, deep vessel density and FD decreased, the area and the form of FAZ remained unchanged in HM subjects comparing with normal subjects. While retina superficial, deep vessel density and FD decreased more, the area of FAZ reduced, the form of FAZ tend to be irregular in mCNV.

Objective:To observe the clinical efficacy of half-dose photodynamic therapy (PDT) in the treatment of acute central serous chorioretinopathy (CSC) by enhanced deep imaging-OCT (EDI-OCT).Methods:A retrospective case study. From September 2015 to November 2018, 100 patients with acute CSC who received half-dose PDT in Hangzhou Branch of the Eye Optometry Hospital of Wenzhou Medical University were included in the study. Among 100 patients, 69 patients were males and 31 patients were females; the average age was 49.63±7.97 years; the average duration of disease was 2.19±0.71 months. All patients underwent BCVA, EDI-OCT, FFA, ICGA and other examinations. BCVA was used on the international standard visual acuity chart and converted to logMAR visual acuity records. Before treatment, the average logMAR BCVA was 0.29±0.19, the average macular foveal retinal thickness (CMT) was 370.59±134.98 μm, and the average macular subfoveal choroidal thickness (SFCT) was 366.93±86.95 μm. All patients were treated with half-dose PDT. We compared the changes of BCVA, CMT, SFCT, and subretinal fluid (SRF) of the eye before treatment and 2 weeks after treatment and 1, 3, and 6 months. Pearson correlation analysis method was used to analyze the correlation between BCVA and baseline BCVA, CMT, SFCT after 6 months of treatment.Results:Six months after treatment, SRF was completely absorbed in 98 eyes, with an effective rate of 98.0%. Compared with before treatment, 2 weeks and 1, 3, and 6 months after treatment, the BCVA of the eye significantly increased ( F=66.493, P<0.001), and CMT and SFCT significantly decreased ( F=134.625, 30.394; P<0.001,<0.001). The results of Pearson correlation analysis showed that BCVA was positively correlated with baseline BCVA 6 months after treatment ( r=0.529, P<0.001), and there was no significant correlation with CMT and SFCT. There were no serious complications related to treatment during the follow-up period. Conclusions:Half-dose PDT can effectively increase BCVA in a short period of time for acute CSC. EDI-OCT can observe that CMT, SFCT and SRF absorption are significantly reduced after treatment.

Objective:To compare the effects of Dynesys dynamic fixation and fusion fixation on the sagittal parameters of lumbar degenerative diseases of 2-4 segments.Methods:A total of 45 patients with 2-4 segments lumbar degenerative diseases, who underwent decompression and internal fixation in our hospital from March 2014 to July 2019, were retrospectively analyzed. There were 23 males and 22 females with an average age of 55±18 years (range 15-86 years). Dynesys dynamic fixation was performed in 22 cases and fusion fixation in 23 cases. Lumbar anteroposterior, flexion and extension lateral and standing full-length spine radiographs were taken before surgery, 2 weeks postoperatively, 3 months postoperatively and at the last follow-up. Sagittal alignment parameters were measured on those radiographs and compared between two groups. Lumbar parameters included lumbar lordosis (LL), lordosis of the fixed segments (LFx), lordosis of the adjacent level to fixed segment (LAdj) and lumbar range of motion (ROM). Pelvic parameters included pelvic incidence (PI), pelvic tilt (PT) and sacral slope (SS).Results:All cases were followed up for 12-50 months with an average of 18.5 months. There was no significant intra- and inter-group difference of LL value between Dynesys dynamic fixation and fusion fixation group before and after surgery ( P>0.05). The overall difference of LFx between groups was not statistically significant ( F=0.700, P=0.406). There was statistically significant variance between time points ( F=7.960, P<0.001) and an interaction effect between group and time ( F=3.940, P=0.006). The LFx of the fusion fixation group was more than that of the dynamic fixation group at 3 months postoperatively and at the last follow-up ( P<0.05). The difference of LAdj value between the two groups was not statistically significant ( F=0.520, P=0.476), while the difference between time points was significant ( F=4.810, P=0.002) with interaction effect between group and time ( F=3.560, P=0.010). Postoperative ROM values of the two groups showed statistically significant differences in group effect, time effect and interaction effect, respectively ( F=4.770, P=0.034; F=18.510, P<0.001; F=5.940, P=0.002). Dynamic fixation group's ROM was more than that of the fusion fixation group at 3 months postoperatively and at the last follow-up ( P<0.05). There was no significant difference of PI between groups and between different time points ( F=0.580, P=0.451; F=0.750; P=0.477). There was no interaction effect in PI between group and time ( F=0.120, P=0.886). There was no significant difference of PT and SS between two groups ( F=0.320, P=0.576; F=0.020, P=0.901). Both time effect and interaction effect were statistically significant ( P<0.05). One complication of unilateral S 1 screw loosening happened in one patient with dynamic fixed at 2 years after surgery. There was no complication in the fusion fixation group during the follow-up. Conclusion:Both Dynesys dynamic fixation and fusion fixation can effectively reconstruct the lumbar lordosis. The fixed segments' lordosis of Dynesys was less than that of fusion fixation from 3 months after operation. Theoretically, the similar increasing trends of LAdj after fixation implied that the degeneration of adjacent segments may occur in both fixation systems. The dynamic fixation can retain more ROM than the fusion fixation. Both two fixation systems have similar influence on the pelvic parameters.

[Abstract] Objective: To explore the effect of circ_0001429 on proliferation and apoptosis of bladder cancer cells by regulating miR-139-5p/TGF-interacting factor 1（TGIF1）axis. Methods: The expression of circ_0001429 in bladder cancer cell lines SW780, T24, 5637 and human bladder epithelial SV-HUC-1 cells were detected by RT-qPCR. Targeted regulatory relationship between circ_0001429 and miR-139-5p as well as miR-139-5p and TGIF1 was measured by Dual luciferase reporter gene assay. T24 cells were divided into NC group, sh-circ_0001429 group, miR-139-5p mimics group, sh-TGIF1 group, pcDNA-circ_0001429+sh-TGIF1 group, miR-139-5p mimics+pcDNA-TGIF1 group and sh-circ_0001429+miR-139-5p inhibitor group. Western blotting was used to detect the expression level of TGIF1 in each group. CCK-8 method, Transwell experiment and Flow cytometry were used to detect the effects of circ_ 0001429, miR-139-5p and TGIF1 on proliferation, invasion, migration and apoptosis of T24 cells, respectively. Results: Circ_0001429 was highly expressed in three bladder cancer cell lines (P<0.01). Knockdown of circ_0001429 significantly inhibited proliferation, invasion and migration of T24 cells while promoted the level of cell apoptosis (P<0.05 or P<0.01). The results of Dual luciferase reporter gene assayconfirmedthatthereisatargetingrelationshipbetweencirc_0001429andmiR-139-5p as well as between miR-139-5p and TGIF1. Overexpression of miR-139-5p significantly inhibited the proliferation, invasion and migration of T24 cells while promoted the level of cell apoptosis (all P<0.01). Recovery experiments further confirmed that the competitive binding of circ_0001429 and TGIF1 to miR-139-5p promoted the proliferation, invasion and migration of T24 cells while inhibited the level of cell apoptosis (all P<0.01). Conclusion: Circ_0001429 promotes proliferation, invasion and migration and inhibits apoptosis of bladder cancer T24 cells by competing with TGIF1 to bind to miR-139-5p.