1.Experimental study to construct the three-dimensional scaffold of rabbit cartilage-shedding cells and evaluate their compatibility with stem cells
Juju XU ; Yanyan XIE ; Zhiyi GUO ; Yukai MA ; Linhong WANG ; Meirong YANG ; Zhaoling DENG ; Baolai HUA ; Zhenyu YAN
Acta Laboratorium Animalis Scientia Sinica 2024;32(8):1012-1022
Objective To prepare decellularized scaffolds from rabbit cartilage at various concentrations and assess their physicochemical properties and compatibility with stem cells to provide an experimental basis for cartilage repair.Methods Bone marrow mesenchymal stem cells(BMSCs)were cultured using the Percoll density gradient separation method,and this was followed by flow cytometric analysis and testing of their osteogenic and chondrogenic differentiation capabilities.Cartilage pieces were excised from rabbit knees and hip joints and subjected to physical crushing,repeated freeze-thaw cycles,and mixed enzymatic digestion for decellularization.To compare and observe the physicochemical properties of the decellularized scaffolds at different concentrations,three groups of scaffolds(labelwd A,B,and C)were designed with concentrations of 100%,50%and 30%,with three replicates each.Third-generation PKH26-labeled BMSCs were seeded onto optimally concentrated scaffolds and cultured for 1 week to observe cell growth.Results Flow cytometry detected BMSC surface antigens with positive expression of CD44 and CD90 and negative expression of CD45.Osteogenic induction stained with alizarin red showed red calcific nodules,and chondrogenic induction stained with alcian blue showed blue cartilaginous nodules.No apparent cell morphology was observed in the three groups of scaffolds stained with hematoxylin-eosin,and toluidine blue.There was a significant difference in DNA concentration between decellularized samples and non-decellularized scaffolds(P<0.05).The content of glycosaminoglycans was slightly lower than the normal values.Significant differences were observed between the three groups of scaffolds in terms of pore size,water absorption,porosity,tensile strength,and Young's modulus(P<0.05).After co-cultivation of stem cells with the scaffolds,cell adhesion was found to be good.Conclusions Percoll density gradient separation can obtain high-purity rabbit BMSCs,and the mixed decellularization method is superior.Group B scaffolds were the most suitable for tissue-engineered cartilage repair.BMSCs cultured in vitro grew well on Group B scaffolds.
2.Clinical analysis of congenital coronary artery fistula in 71 children
Yue LIU ; Fen LI ; Lijun FU ; Tingliang LIU ; Ying GUO ; Wei GAO ; Meirong HUANG ; Hao ZHANG ; Jie SHEN
Chinese Journal of Applied Clinical Pediatrics 2022;37(2):94-97
Objective:To explore the clinical characteristics, therapeutic efficacy and prognosis of congenital coronary artery fistula (CAF) in children.Methods:Clinical data of 71 pediatric patients diagnosed with congenital CAF at Department of Cardiology and Department of Cardiac Surgery, Shanghai Children′s Medical Center, Shanghai Jiao Tong University School of Medicine from January 2013 to June 2019 were retrospectively analyzed.The median age was 2.2 years (0.1-14.0 years), and the median body weight was 18.3 kg (3.2-55.8 kg), including 37 males (52.1%) and 34 females (47.9%). They were divided into the transcatheter closure group (30 cases) and surgical repair group (41 cases). The therapeutic effect and follow-up data of the 2 groups were compared by the Fisher′ s exact test. Results:Among the 71 congenital CAF children, 70 had heart murmurs, 2 had chest tightness after activity, and 5 were prone to recurrent respiratory tract infection.Transcatheter closure and surgical repair were successfully performed in 22/30 (73.3%) and 41/41 (100.0%) cases with a statistically significant difference ( P=0.001). However, in the surgical repair group, 1 (2.4%) case died after operation and 2(4.9%) needed further transcatheter closure due to large residual shunt during the follow-up period.At the last follow-up, there were 2 cases with minimal or small residual shunt in both groups ( P=0.567). There were 2/71(2.8%) cases suffering from postoperative thrombosis. Conclusions:Cardiac murmur is the main sign of congenital CAF in children, and some of them may have frequent respiratory tract infection due to increased lung blood caused by a large amount of shunt.A few children have chest tightness and chest pain due to myocardial ischemia caused by coronary steal.Both percutaneous closure and surgical repair are safe and effective with few complications.
3.Effect and mechanism of astaxanthin on acute kidney injury in rats with full-thickness burns
Meirong YU ; Songxue GUO ; Ronghua JIN ; Chuangang YOU ; Xingang WANG ; Chunmao HAN
Chinese Journal of Burns 2020;36(11):1050-1059
Objective:To explore the effect and mechanism of astaxanthin on acute kidney injury in rats with full-thickness burns.Methods:Forty-eight male Sprague Dawley rats of 8 to 10 weeks were divided into sham injury group, simple burn group, burn+ vehicle group, burn+ low-dose astaxanthin group, burn+ medium-dose astaxanthin group, and burn+ high-dose astaxanthin group according to the random number table, with 8 rats in each group. The back skin of rats in sham injury group were immersed in warm water of 20 ℃ for 15 s to simulate burn injury, and the back skin of rats in the other 5 groups were immersed in boiled water of 100 ℃ for 15 s to inflict full-thickness burn of 30% total body surface area. Fluid resuscitation was performed in rats in the 5 groups except of sham injury group immediately and 6 h after injury. At 30 min after injury, the rats in sham injury group and simple burn group were injected with 1 mL/kg normal saline via tail vein, rats in burn+ vehicle group were injected with 1 mL/kg astaxanthin solvent via tail vein, and rats in burn+ low-dose astaxanthin group, burn+ medium-dose astaxanthin group, and burn+ high-dose astaxanthin group were respectively injected with 5, 10, 20 mg/kg astaxanthin solution of 5, 10, 20 mg/mL via tail vein. The renal tissue was collected at post injury hour (PIH) 48, and hematoxylin eosin staining was used for histopathological observation and renal tubular injury score. At PIH 48, the venous blood was collected for detecting serum creatinine level through blood biochemical analyzer, and blood urea nitrogen (BUN) level was detected by enzyme-linked immunosorbent assay. The renal tissue was collected to detect the mRNA expressions of myeloperoxidase (MPO), interleukin-1β (IL-1β), and IL-6 by real-time fluorescent quantitative reverse transcription polymerase chain reaction method, and the protein expressions of Toll like receptor 4 (TLR4), phosphorylated nuclear factor kappa B (p-NF-кB) p65, and heme oxygenase 1 (HO-1) were detected by Western blotting. Besides, the expression of HO-1 in renal tissue was detected by immunofluorescence method. Data were statistically analyzed with Kruskal-Wallis H test, Dunn-Sidák correction, one-way analysis of variance, and Bonferroni method. Results:(1) At PIH 48, there were no inflammatory cell infiltrating and degeneration or necrosis of cells in renal tissue of rats in sham injury group, and the structure of renal tubules was intact. The renal tubules of burn rats in each group showed injury manifestation of separation between epithelial cell and basement membrane, and vacuole cells and lysate protein aggregation. The injury degree of renal tissue of rats in burn+ high-dose astaxanthin group was obviously decreased compared with that in simple burn group. (2) At PIH 48, compared with that of sham injury group, the renal tubular damage scores of rats in simple burn group, burn+ vehicle group, burn+ low-dose astaxanthin group, and burn+ medium-dose astaxanthin group were significantly increased ( P<0.05 or P<0.01). Compared with those of simple burn group and burn+ vehicle group, the renal tubular damage scores of rats in burn+ medium-dose astaxanthin group and burn+ high-dose astaxanthin group were significantly decreased ( P<0.05 or P<0.01). Compared with that of burn+ low-dose astaxanthin group, the renal tubular damage score of rats in burn+ high-dose astaxanthin group was significantly decreased ( P<0.01). (3) At PIH 48, the level of serum creatinine of rats in sham injury group was (2.42±0.06) mg/L, which was significantly lower than (6.11±0.11), (6.48±0.08), (5.79±0.09), (4.03±0.12) mg/L of simple burn group, burn+ vehicle group, burn+ low-dose astaxanthin group, and burn+ medium-dose astaxanthin group ( P<0.05 or P<0.01). The level of BUN of rats was (21.9±1.3) mmol/L in sham injury group, significantly lower than (32.1±7.4) mmol/L of simple burn group and (30.2±4.8) mmol/L of burn+ vehicle group ( P<0.05 or P<0.01). At PIH 48, compared with those of simple burn group and burn+ vehicle group, the levels of serum creatinine and BUN of (16.0±2.9) mmol/L in burn+ medium-dose astaxanthin group, serum creatinine of (3.02±0.08) mg/L and BUN of (14.5±2.9) mmol/L in burn+ high-dose astaxanthin group, and serum creatinine of (22.8±5.5) mmol/L of rats in burn+ low-dose astaxanthin group were significantly decreased ( P<0.05 or P<0.01). At PIH 48, compared with those of burn+ low-dose astaxanthin group, the levels of serum creatinine and BUN of burn+ high-dose astaxanthin group and serum creatinine of burn+ medium-dose group were obviously decreased ( P<0.05 or P< 0.01). (4) At PIH 48, compared with those of sham injury group, the mRNA expressions of MPO, IL-1β, and IL-6 in renal tissue of rats in simple burn group, burn+ vehicle group, burn+ low-dose astaxanthin group, and burn+ medium dose astaxanthin group, and the mRNA expressions of IL-1β and IL-6 in renal tissue of rats in burn+ high-dose astaxanthin group were obviously increased ( P<0.01). Compared with those of simple burn group and burn+ vehicle group, the mRNA expressions of MPO, IL-1β, and IL-6 in renal tissue of rats were significantly decreased in burn+ low-dose astaxanthin group, burn+ medium-dose astaxanthin group, and burn+ high-dose astaxanthin group ( P<0.01). Compared with those of burn+ low-dose astaxanthin group, the mRNA expressions of MPO, IL-1β, and IL-6 in renal tissue of rats were significantly decreased in burn+ medium-dose astaxanthin group and burn+ high-dose astaxanthin group ( P<0.01). The mRNA expressions of MPO, IL-1β, and IL-6 in renal tissue of rats in burn+ high-dose astaxanthin group were significantly decreased compared with those of burn+ medium-dose astaxanthin group ( P<0.01). (5) At PIH 48 h, compared with those of sham injury group, the protein expressions of TLR4 and p-NF-кB p65 in renal tissue of rats in simple burn group, burn+ vehicle group, burn+ low-dose astaxanthin group, and burn+ high-dose astaxanthin group were obviously increased ( P<0.01). Compared with those of simple burn group, the protein expressions of TLR4 and p-NF-кB p65 in renal tissue of rats in burn+ low-dose astaxanthin group, burn+ medium dose astaxanthin group, and burn+ high-dose astaxanthin group were significantly decreased ( P<0.01). (6) The results of Western blotting combined with immunofluorescence method showed that compared with that of sham injury group, the protein expression of HO-1 in renal tissue of rats in burn+ vehicle group, burn+ low-dose astaxanthin group, burn+ medium-dose astaxanthin group, and burn+ high-dose astaxanthin group were significantly increased at PIH 48 ( P<0.01), and the protein expression of HO-1 in renal tissue of rats in burn+ medium-dose astaxanthin group and burn+ high-dose astaxanthin group was significantly increased compared with that of simple burn group ( P<0.01). Conclusions:Astaxanthin can attenuate the structural damage and functional decline of renal tissue and regulate the release of injury-related inflammatory factors, thus to protect the rats from acute kidney injury after burn. The HO-1/TLR4/NF-кB signaling pathway is the main regulatory mechanism of astaxanthin to achieve anti-inflammation-based renoprotection.
4. Clinical analysis of Staphylococcus aureus infective endocarditis in children
Xinyi XU ; Ying GUO ; Tingliang LIU ; Lijun FU ; Jie SHEN ; Xu ZHANG ; Jinjin WU ; Wei GAO ; Fen LI ; Meirong HUANG
Chinese Journal of Applied Clinical Pediatrics 2019;34(22):1707-1710
Objective:
To describe the clinical characteristics, treatment and outcomes of
5. Clinical characteristics of anomalous origin of the left coronary artery from the pulmonary artery in 91 children
Xian MEI ; Fen LI ; Lijun FU ; Haibo ZHANG ; Yuqi ZHANG ; Wei GAO ; Meirong HUANG ; Tingliang LIU ; Ying GUO ; Jie SHEN
Chinese Journal of Pediatrics 2019;57(8):614-619
Objective:
To explore the clinical manifestations, diagnosis, treatment and prognosis of anomalous origin of the left coronary artery from the pulmonary artery (ALCAPA) .
Methods:
A retrospective study identified 91 patients diagnosed with ALCAPA at Shanghai Children′s Medical Center from March 2010 to August 2017. According to the left ventricular ejection fraction (LVEF) at the time of consultation, patients were divided into the cardiac insufficiency group (
6.Clinical characteristics of anomalous origin of the left coronary artery from the pulmonary artery in 91 children
Xian MEI ; Fen LI ; Lijun FU ; Haibo ZHANG ; Yuqi ZHANG ; Wei GAO ; Meirong HUANG ; Tingliang LIU ; Ying GUO ; Jie SHEN
Chinese Journal of Pediatrics 2019;57(8):614-619
Objective To explore the clinical manifestations, diagnosis, treatment and prognosis of anomalous origin of the left coronary artery from the pulmonary artery (ALCAPA). Methods A retrospective study identified 91 patients diagnosed with ALCAPA at Shanghai Children′s Medical Center from March 2010 to August 2017. According to the left ventricular ejection fraction (LVEF) at the time of consultation, patients were divided into the cardiac insufficiency group (n=54) and the normal cardiac function group (n=37). Clinical features (age of onset, clinical performance, etc) and auxiliary examinations (electrocardiogram, echocardiography, etc) between the two groups were compared using a t‐test and a Chi‐square test. Prognostic factors were analyzed by an ordered logistic regression and a Pearson correlation coefficient. Results (1) The age of diagnosis of patients in the cardiac insufficiency group who were usually misdiagnosed as cardiomyopathy was (10.0±2.6) months (20/54), whereas the age of diagnosis of patients in the normal cardiac function group who were usually misdiagnosed as valvular diseases was (40.0 ± 7.8) months (4/37). According to the pathophysiological mechanism, forty of the 54 (74%) patients in the cardiac insufficiency group were infantile type, and 78% patients (29/37) in the normal cardiac function group were adult type. (2) Preoperative electrocardiogram showed the deep Q wave in lead I occurred more frequently in the cardiac insufficiency group than in the normal cardiac function group (28/54 vs. 11/37, χ2=4.388, P=0.036). (3) Twenty patients died in the cardiac insufficiency group including 12 patients who died from postoperative cardiac pump failure and 8 children who did not undergo surgery due to poor prognosis and died from other reasons. There was no death in the normal cardiac function group. (4) Preoperative LVEF was the unique risk factor affecting prognosis (F=16.872, P=0.005). The preoperative LVEF was significantly lower than the postoperative LVEF ((37±11)% vs. (45±14)%, t=3.614, P=0.001) in the cardiac insufficiency group. During the follow‐up period, 6 patients in the cardiac insufficiency group still presented with postoperative cardiac dysfunction, and the patients in the normal cardiac function group still had normal cardiac function. Conclusions Preoperative LVEF was the unique risk factor affecting prognosis of ALCAPA. Patients with infantile type ALCAPA and preoperative cardiac insufficiency should receive long‐term follow‐up treatment.
7.Clinical study of 62 cases of anomalous left coronary artery from the pulmonary artery
Xu ZHANG ; Meirong HUANG ; Tingliang LIU ; Wei GAO ; Fen LI ; Ying GUO ; Lijun FU ; Xinyi XU
Journal of Clinical Pediatrics 2018;36(6):438-442
Objective To explore the clinical manifestation, treatment and prognosis of anomalous left coronary artery from the pulmonary artery (ALCAPA). Method The clinical data of patients with ALCAPA from January 2011 to September 2016 were retrospectively analyzed. All the patients were divided into infant group (<12 months) and child group (≥12 months). Results In the 62 patients (38 infants and 24 children) with ALCAPA, the median ΔLVDD (actually measured LVDD – the normal upper limit of LVDD in this age group) of infant group and child group was 13.6 (8.4~17.5) mm and 8.5 (3.3~13.7) mm respectively. The mean LVEF of the infant group was 40.6±2.4 %, lower than that of the children group (59.0±2.9) %, and the difference was statistically significant (P<0.01). Sixty children were treated with surgery. The mean cardiopulmonary bypass duration of infant group (137.1±8.4 min) was longer than that of the children group (105.9±6.6 min), and the difference was statistically significant (P=0.010). The median mechanical ventilation time after operation in infant group (6 d, 3~7 d), was longer than that in the children group (2 d, 1~4 d), and the difference was statistically significant (P<0.01). The median follow-up time of 60 patients was 38 months. Six patients died within 1 month after surgery (5 cases in the infant group and 1 in the children group), and there were no deaths more than 1 month after surgery. In 22 cases, the flow rate of pulmonary valve increased rapidly within 1 week after operation (>1.8 m/s). Conclusion ALCAPA is easily misdiagnosed as dilated cardiomyopathy. The clinical manifestations of infants were more severe than those of children. The mortality was high within 1 month after surgery, and the long-term prognosis was better.
8.Advances in the research of the relationship between wound temperature and wound healing
Lingya ZHU ; Songxue GUO ; Pan WU ; Qiong LI ; Jonkee HO ; Meirong YU ; Tingting WENG ; Chunmao HAN
Chinese Journal of Burns 2018;34(11):829-832
There are many factors that may affect the microenvironment of acute and chronic wounds.This article reviews the relationship between temperature factor in the external microenvironment of wound surface and wound healing.The temperature changes in different types and stages of wounds are closely related to the wound healing status.Therefore,wound temperature monitoring provides timely,reliable,and non-invasive method in the evaluation of wound status.As low temperature affects the physiological state of wound,relieving the low temperature state and maintaining normal temperature of the microenvironment of wound can promote wound healing.Further research is needed on the wound repair related effector cell proliferation and the mechanism of regulatory function to determine the optimal constant temperature and heat treatment duration needed for wound healing.
9.Diagnostic value of invasive cardiac catheterization for constrictive pericarditis and restrictive cardiomyopathy
Ying GUO ; Wei GAO ; Lijun FU ; Meirong HUANG ; Tingliang LIU ; Xinyi XU ; Xu ZHANG
Chinese Journal of Applied Clinical Pediatrics 2017;32(1):34-37
Objective To evaluate the diagnostic value of invasive cardiac catheterization for restrictive cardiomyopathy (RCM) and constrictive pericarditis (CP).Methods Twenty-seven children with CP or RCM hospitalized in Department of Cardiology,Shanghai Children's Medical Center,School of Medicine,Shanghai Jiaotong University from February 2002 to December 2015,were selected,including 10 patients who had been surgically documented CP and 17 patients with RCM who underwent cardiac catheterization.Intracardiac pressure waveforms were recorded and all the measurement indexes of pressure of all the patients were analyzed.The changes of cardiac pressure under deep breathing in 9 patients with local anesthesia was recorded.Results There were significant differences in pulmonary artery pressure,difference between left ventricular end-diastolic pressure (LVEDP) and right ventricular end-diastolic pressure (RVEDP),and the ratio of RVEDP/right ventricular systolic pressure between group RCM and group CP [(50.2 ± 12.0) mmHg(1 mmHg =0.133 kPa) vs.(38.1 ±6.8) mmHg,(7.8±5.5) mmHgvs.(1.8 ±4.7) mmHg,0.27 ±0.10vs.0.45 ± 0.20,respectively;t =2.912,2.787,2.418,all P < 0.05].However,there was overlapping for these criteria,and the predictive sensitivity of any of the criteria was less than 66.7%.In patients with CP,reciprocal changes in the filling between right ventricle and left ventricle occurred during respiration.In patients with RCM,the right ventricle and left ventricle pressures moved concordantly with respiration.The systolic area index was greater in group CP than that in group RCM (1.20 ± 0.03 vs.0.70 ± 0.14),and the difference was significant (t =6.152,P < 0.01).The systolic area index had a sensitivity of 100.0% and a predictive accuracy of 100.0% for the identification of patients with surgically proven CP.Conclusions Measurements in catheterization,especially the ratio of right ventricular to left ventricular systolic area during inspiration and expiration is a reliable catheterization criterion for differentiating CP from RCM.
10.Treatment of infective endocarditis in 29 children with Linezolid
Xinyi XU ; Meirong HUANG ; Ying GUO ; Tingliang LIU ; Lijun FU ; Xu ZHANG ; Jinjin WU ; Wei GAO ; Fen LI ; Jie SHEN
Chinese Journal of Applied Clinical Pediatrics 2017;32(13):982-985
Objective To explore the effectiveness and safety of Linezolid (LIZ) in treating children with infective endocarditis (IE).Methods The clinical data of 112 children with IE and treated in the Shanghai Children's Medical Center of Shanghai Jiao Tong University School of Medicine from August 2008 to September 2015 were retrospectively analyzed.There were 64 boys and 48 girls,and the age of IE onset ranged from 1 month to 17 years [(6.0 ± 4.8) years].Twenty-nine patients received LIZ treatment > 7 days (LIZ treatment group),including 21 males and 8 females,and the age ranged from 5 months to 15 years [(6.9 ±5.2) years].The remaining 83 patients were identified without LIZ treatment(non-LIZ treatment group),including 43 boys and 40 girls,and the age ranged from 1 month to 17 years [(5.7 ±4.7) years].The etiological results,curative effect and adverse reactions of the LIZ treatment group were observed.Results Among the LIZ treatment group,22 cases had congenital heart disease and 1 case had intravenous catheter.There were 10 cases with infection of staphylococcus aureus,6 cases with coagulase negative staphylococcus,5 cases with oral streptococci and Streptococcus bovis group,3 cases with streptococcus pneumonia,2 cases with enterococcus faecium and 3 cases with negative blood culture results.All cases of the LIZ treatment group received Vancomycin therapy at first,LIZ was given when the Vancomycin therapy failed(16 cases with temperature reiteration,1 case inadequate microbiological response),Vancomycin intolerance (6 neutropenia,2 renal toxicity,2 allergy),and oral maintenance therapy (2 cases).The duration of LIZ treatment ranged from 9 to 135 days [(39.2 ±27.2) days].Three patients (10.3%) had adverse effects during LIZ treatment,1 case with severe digestive symptoms after treatment,1 case with teeth discoloration,1 case with the indicators decreasing by 2 routine blood test.Meanwhile,16 cases of 85 patients (18.8%) had side effects during Vancomycin treatment,in which 7 cases with neutropenia,6 cases with rash and 3 cases with renal insufficiency.But there were no significant differences in adverse effects between LIZ and Vancomycin treatment (x2 =1.l19,P >0.05).Twenty-five cases were cured (86.2%) and 2 cases dead (6.9%) in the LIZ treatment group.And no significant difference was found in cure rate,or mortality between LIZ treatment group and non-LIZ treatment group (86.2% vs.77.1%,x2 =1.090;6.9% vs.8.4%,x2 =0.069,all P > 0.05) at 6 to 84 (23.9 ± 19.1) months follow-ups.Conclusions LIZ can be used to deal with Vancomycin failure and IE caused by gram positive coccus.LIZ was generally well tolerated in patients with IE.It may be useful in cases of IE complicated by brain abscesses for the good distribution.It is 100% orally bioavailable,allowing oral administration for outpatients.

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