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Objective:To explore the morbidity features and therapeutic outcomes of rejections in pediatric kidney transplantation (KT) recipients.Methods:Between January 2013 and June 2022, 360 children undergoing KT were recruited.The relevant clinical data were collected for examining the morbidity features and therapeutic outcomes of rejections.The serum levels of creatinine were compared among groups by non-parametric rank test.And Kaplan-Meier and Log-rank methods were employed for examining the incidence of rejection and comparing mortality-censored graft survival rates among patients with different times of rejection.Results:A total of 58 recipients had 82 incidents of rejection with a cumulative incidence of 6.3%, 9.2% and 11.3% at 3/6/12 months respectively.Among 50 incidents of biopsy-proved rejections, the types were T cell-mediated rejection [TCMR, 42.0%(21/50)], antibody-mediated rejection [20.0%(10/50), ABMR] and mixed rejection [38.0%(19/50)].Among 58 incidents of initial rejection, 69% had maintained graft function (MGF) and 31% impaired graft function (IGF) after anti-rejection regimens.Among 80.8%, 85.7% and 75% of recipients with clinical rejection, ABMR or borderline rejection while 36.4% in TCMR patients had MGF.Fifteen kidney allografts lost function in 58 recipients with rejection.Five-year death-censored graft survival was significantly lower in patients with two or more incidents of rejection (30.5%, 95% CI: 12.3%-75.4%) than in those without rejection (92.9%, 95% CI: 89.3%-96.6%) ( P<0.000 1) or with only one rejection (82.9%, 95% CI: 65.9%-100%)( P<0.001). Conclusions:The rejection rate remains high in KT children and it affects graft survival.And TCMR is more likely to cause impaired graft function.Recurrent rejections have a more pronounced impact upon graft survival.
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Objective:To investigate the change of syndesmotic volume via CT scan in Danis-Weber B type ankle fracture, aiming to provide a non-invasive diagnostic method for Danis-Weber B type ankle fracture combined with syndesmotic injury.Methods:Retrospective analysis was performed on 48 patients with Danis-Weber B type ankle fractures in Shanghai Sixth People′s Hospital Affiliated to Shanghai Jiao Tong University School of Medicine from January 2018 to March 2022, including 30 males and 18 females, aging (43.71±19.41) years. Ankle CT scans and three-dimensional reconstructions were performed before surgery and at the last follow-up, measuring the syndesmotic volume between the tibiotalar joint and 1, 3 and 5 cm above the ankle and compared with each other. Intraoperative exploration was used as the gold standard for the syndesmotic injury. Based on the results of exploration of the syndesmotic injury, patients were divided into simple ankle fracture group (fracture group, 25 cases) and ankle fracture combined with syndesmotic injury group (fracture + ligament group, 23 cases). The sensitivity and specificity of this diagnostic method were statistically evaluated. The assessments at the last follow-up were performed to value the clinical effect of surgery. The metric data conforming to the normal distribution were expressed as mean ± standard deviation ( ± s), and the t-tests were used for comparison between groups. The measurement data of skewed distribution were expressed as quartile M( Q1, Q3), and nonparametric tests were used for intergroup comparison. Counting data were expressed as number of cases and percentage (%), and Chi-square test were used for intergroup comparison. Results:In the fracture group, there were no statistically significant differences of the syndesmotic volume at 1, 3, and 5 cm above the ankle joint before surgery and at the last follow-up ( P=0.219, 0.269, 0.103). On the contrary, the volume above were statistically significant in the fracture + ligament group ( P<0.001). There were statistically significant differences in syndesmotic volume between the two groups at 1, 3, and 5 cm above the ankle joint preoperatively ( P=0.005, 0.004, 0.038). By contrast, there were no statistical differences between the two groups postoperatively ( P=0.082, 0.155, 0.249). For the sensitivity and specificity of Danis-Weber B type ankle fractures combined with syndesmotic injury, they were 92% and 67% at 1 cm above the ankle joint, 69% and 87% at 3 cm above the ankle joint, and 62% and 87% at 5 cm above the ankle joint, respectively. The last follow-up clinical function score indicated a good surgical outcome. Conclusions:Volumetric measurement via CT scan is one of the diagnostic methods for evaluating Danis-Weber B type ankle fracture combined with syndesmotic injurys. Open resection internal fixation combined with elastic fixation of the ankle fracture combined with syndesmotic injurys can significantly reduce the lower tibiofibular volume, and the efficacy is definite.
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Objective:To investigate the efficacy of osteochondral fragment fixation using bioabsorbable pins for Hepple Ⅱ osteochondral lesions of the talus (OLT) in adolescents.Methods:Retrospective case analysis was used. The clinical data and follow-up results of 13 adolescent patients (13 feet) with Hepple Ⅱ OLT were all treated with osteochondral fragment fixation using bioabsorbable pins admitted to Shanghai Sixth People′s Hospital Affiliated to Shanghai Jiao Tong University School of Medicine from January 2017 to December 2021 were retrospectively analyzed. There were 7 males and 6 females, with 13 right feet. The age was (14.85±2.23) years old, ranged from 12 to 18 years old. According to the American orthopedic foot and ankle society (AOFAS) ankle-hindfoot score, visual analogue scale (VAS) and SF-36 score before operation and at the last follow-up were used to evaluate the efficacy and function of the patients. Measurement data with normal distribution were represented as mean ± standard deviation( ± s), and the comparison between groups was conducted using the t-test; The mearsurement data with skewness distribution were expressed by M( Q1, Q3), and rank-sum test was used for inter-group comparison. Results:Thirteen adolescent patients (13 feet) with Hepple Ⅱ OLT underwent surgery successfully and were followed up for (25.54±9.95) months. All wounds healed by first intention, and no complications such as wound infection and delayed healing occurred. Preoperative AOFAS ankle-posterior foot score, VAS and SF-36 score were 58.62±3.55, 7.00 (6.50, 8.00) and 68.38±4.81, respectively. At the last follow-up, the scores were 97.38±2.73, 1.00 (0.00, 1.00), 91.15±4.28, respectively, and the results were significantly improved at the last follow-up, with the difference between the two groups statistically significant( P<0.05). Conclusion:Osteochondral fragment fixation using bioabsorbable pins which can promote cartilage repair, significantly improve symptoms, and achieve better clinical satisfaction with fewer complications, is a safe and effective surgical treatment option for Hepple Ⅱ OLT in adolescents with satisfactory short-term clinical outcomes.
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ObjectiveTo explore the mechanism of Buyang Huanwutang combined with bone marrow mesenchymal stem cell （BMSC） transplantation in the treatment of spinal cord injury （SCI）. MethodDifferent concentrations （12.5， 25， 50 g·kg-1） of Buyang Huanwutang were administrated to rats by gavage. The spinal cord function of rats was measured by modified Tarlov score， and the most suitable concentration of Buyang Huanwutang was screened out. SD rats were then divided into 6 groups， namely， the sham operation group （gavage of equal amount of normal saline）， the model group （gavage of equal amount of normal saline）， the Buyang Huanwutang group （gavage of 25 g·kg-1 Buyang Huanwutang）， the BMSC transplantation group （tail vein injection of BMSCs 1 mL）， the Buyang Huanwutang+BMSC group （gavage of 25 g·kg-1 Buyang Huanwutang and tail vein injection of BMSCs 1 mL）， the Buyang Huanwutang+BMSC+LY294002 group （gavage of 25 g·kg-1 Buyang Huanwutang and tail vein injection of BMSCs 1 mL and 40 mg·kg-1 LY294002）， with 10 rats in each group. The spinal cord function was measured by the modified Tarlov score， inclined plate test， and latency of cortical somatosensory evoked potential. Immunohistochemistry was used to detect the number of 5-bromo-2-deoxyuracil nucleoside （Brdu）-labeled positive cells in the spinal cord tissue. The protein expression levels of phosphorylated protein kinase B （p-Akt）， glycoprotein 130 （gp130）， and interleukin-6 （IL-6） in spinal cord were detected by Western blot. ResultAs compared with the sham operation group， the Tarlov score and the critical angle of tilt plane in the model group were significantly decreased （P<0.05）， and the latency of cortical somatosensory evoked potential wave and the protein expression levels of p-Akt， gp130， and IL-6 were significantly increased （P<0.05）. As compared with the model group， the Tarlov score and the critical angle of tilt plane in the sham operation group and each treatment group were significantly increased （P<0.05）， and the latency of cortical somatosensory evoked potential wave and the protein expression levels of p-Akt， gp130， and IL-6 were significantly decreased （P<0.05）. As compared with the BMSC group， the Tarlov score and the critical angle of inclined plane in the Buyang Huanwutang+BMSC group increased （P<0.05）， the latency of cortical somatosensory evoked potential wave and the protein expression levels of p-Akt， gp130， and IL-6 decreased （P<0.05）， and the number of Brdu-labeled positive cells increased 5 weeks after transplantation （P<0.05）. As compared with the Buyang Huanwutang+BMSC group， the Tarlov score and the critical angle of the inclined plane in the Buyang Huanwutang+BMSC+LY294002 group increased （P<0.05）， and the latency of cortical somatosensory evoked potential wave and the protein expression levels of p-Akt， gp130， and IL-6 decreased significantly （P<0.05）. Five weeks after transplantation， the number of Brdu-labeled positive cells increased significantly in the Buyang Huanwutang+BMSC+LY294002 group （P<0.05）. ConclusionBuyang Huanwutang can promote BMSCs migration and restore spinal cord function by inhibiting phosphatidylinositol 3-kinase （PI3K）/protein kinase B （Akt） signal.
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Objective:To conduct a retrospective analysis of efficacy and safety of different conversion schemes of tacrolimus to slow-release dosage forms for recipients in stable phase after renal transplantation to provide rationales for the conversion strategy of tacrolimus.Methods:From January 2020 to June 2020, clinical data were reviewed for 101 kidney transplant recipients converting from common tacrolimus dosage form to tacrolimus sustained-release dosage form during postoperative stable period.There were 62 males and 49 females with an age range of 19 to 69 years.They were divided into two groups according to iso-dose and incremental-dose switching schemes.The common dosage form of tacrolimus was converted into a sustained-release dosage form with different conversion doses, They were divided into two groups of 1∶1 conversion( n=55)and >1∶1 conversion( n=46). The clinical parameters of serum creatinine(Scr), blood urea nitrogen(BUN), alanine aminotransferase(ALT)and aspartate aminotransferase(AST), alkaline phosphatase(ALP), serum albumin(ALB), white blood cell count(WBC), urinary white blood cell(UWBC), hemoglobin(Hb)and fasting blood glucose(Glu)were compared between two groups after conversion. Results:Regarding numerical change trend after switching to tacrolimus sustained-release dosage form, drug dose/variation trend was smaller and blood drug concentration more stabilized.In two subgroups converted by 1∶1 and 1>1 initial dose, change trend of dose/blood concentration in 1∶1 conversion group appeared to be more stable.However, no inter-group difference existed in long-term parameters.Scr was lower at 1 week and 3 months after switching to extended-release dosage form( P<0.05)and BUN was lower at 2 weeks( P<0.05). In addition, at 5 months after conversion, ALT and AST significantly improved as compared with common dosage form( P<0.05). Significant differences existed in urinary WBC(UWBC)at 2/3 weeks( P<0.05). After switching for 2 weeks, hemoglobin significantly improved compared with common dosage form( P<0.05). No significant differences existed in ALP, ALB or Glu at other timepoints and pre-conversion( P>0.05). In 1∶1 switch group, renal function tended to improve.At 2 weeks, BUN was lower than pre-conversion; at 1/3 weeks, Scr was lower than pre-conversion( P<0.05). In addition, there was also a trend of improvement in liver function in 1∶1 conversion group.At 1 week and 5 months, ALT was lower than pre-conversion( P<0.05). However, no significant differences existed in AST, ALB, ALP, Glu, UWBC and serum WBC count at each timepoint between two different dose conversion groups( P>0.05). After conversion, intra-individual variability of tacrolimus trough concentration significantly improved( P<0.05). Conclusions:With the same safety and efficacy as common dosage form, sustained-release dosage form of tacrolimus may improve drug variability of individuals.When converting common dosage form into sustained-release dosage form, individual differences should be considered.While monitoring trough concentrations, proper doses should be adjusted on the basis of various clinical parameters.
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Objective:To explore the diagnosis and treatment of transplanted renal artery stenosis(TRAS)in children.Methods:From January 2016 to August 2021, clinical data of 7 TRAS patients were collected.A definite diagnosis was confirmed by Doppler ultrasound and computed tomography angiography.Results:Patient age was significantly higher than donor age(11.9±3.7 vs 1.0±0.5 years, P<0.001); 5 patients had a widened diameter at stenotic grafted renal artery after intervention(1.98±0.47 vs 4.64±1.19 mm, P=0.002). A reduction in peak systolic flow velocity in stenotic segment of artery(463.3±90.6 vs 183.6±58.9 cm/s, P<0.001)and lower systolic blood pressure(137.2±15.5 vs 129.7±12.3 mmHg, P=0.029)were observed.Resistance index rose(0.38±0.22 vs 0.60±0.03, P=0.063). Significant difference of estimated glomerular filtration rate was observed at Week 4 post-operation as compared with pre-intervention.Two patients developed complications after intervention, including perirenal hematoma and stent-attached thrombus.Two patients were treated conservatively with a gradual increase in blood pressure and three antihypertensive drugs prescribed. Conclusions:Doppler ultrasound should be performed regularly after renal transplantation for detecting TRAS at an early stage in children.Interventional treatment is ideal for severe TRAS to improve perfusion and renal function.Clinicians should pay more attention to complications.
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ObjectiveTo investigate the effect of calycosin-mediated glucoprotein130/Janus kinase/signal transducer and activator of transcription factor （GP130/JAK/STAT） signaling pathway on oxidative injury of astrocytes in spinal cord. MethodAstrocytes in rat spinal cord were isolated and identified by immunofluorescence detection of glial fibrillary acidic protein （GFAP）. The cells were respectively pre-treated with 5， 10， 20 μmol·L-1 calycosin for 12 h， and then 100 μmol·L-1 H2O2 （24 h） was added to induce oxidative injury. Cell counting kit-8 （CCK-8） assay was employed to detect cell proliferation and select the optimal concentration of calycosin. The following experimental groups were designed： control group， model group （100 μmol·L-1 H2O2）， calycosin group （20 μmol·L-1 calycosin）， calycosin + LY294002 group （20 μmol·L-1 calycosin + 10 μmol·L-1 LY294002）， and calycosin + Stattic group （20 μmol·L-1 calycosin + 3 μmol·L-1 Stattic）. CCK-8 assay and immunofluorescence method were used to detect the proliferation of cells and flow cytometry was applied to detect cell apoptosis and cycle. The protein expression of phosphorylated （p）-JAK2， p-STAT3， p-protein kinase B （Akt）， GP130， and interleukin-6 （IL-6） was detected by Western blotting. ResultCompared with the control group， the model group showed low proliferation activity and high apoptosis rate of cells （P<0.05）. Compared with the model group， calycosin （20 μmol·L-1） group displayed high proliferation activity and low apoptosis rate of cells （P<0.05）. Compared with calycosin （20 μmol·L-1） group， both phosphatidylinosirtol-3-kinases （PI3K） inhibitor LY294002 and STAT3 inhibitor Stattic significantly reduced the proliferation activity and increased the apoptosis rate of cells （P<0.05）. The protein expression of p-JAK2， p-STAT3， p-Akt， GP130， and IL-6 in the model group was higher than that in the control group （P<0.05）， and the expression of the above indicators was lower in each treatment group than in the model group （P<0.05）. ConclusionCalycosin can promote the proliferation and inhibit the apoptosis of astrocytes with oxidative injury by inhibiting the phosphorylation of PI3K/Akt pathway and JAK2/STAT3 pathway.
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ObjectiveTo investigate the effect of calycosin-mediated glucoprotein130/Janus kinase/signal transducer and activator of transcription factor （GP130/JAK/STAT） signaling pathway on oxidative injury of astrocytes in spinal cord. MethodAstrocytes in rat spinal cord were isolated and identified by immunofluorescence detection of glial fibrillary acidic protein （GFAP）. The cells were respectively pre-treated with 5， 10， 20 μmol·L-1 calycosin for 12 h， and then 100 μmol·L-1 H2O2 （24 h） was added to induce oxidative injury. Cell counting kit-8 （CCK-8） assay was employed to detect cell proliferation and select the optimal concentration of calycosin. The following experimental groups were designed： control group， model group （100 μmol·L-1 H2O2）， calycosin group （20 μmol·L-1 calycosin）， calycosin + LY294002 group （20 μmol·L-1 calycosin + 10 μmol·L-1 LY294002）， and calycosin + Stattic group （20 μmol·L-1 calycosin + 3 μmol·L-1 Stattic）. CCK-8 assay and immunofluorescence method were used to detect the proliferation of cells and flow cytometry was applied to detect cell apoptosis and cycle. The protein expression of phosphorylated （p）-JAK2， p-STAT3， p-protein kinase B （Akt）， GP130， and interleukin-6 （IL-6） was detected by Western blotting. ResultCompared with the control group， the model group showed low proliferation activity and high apoptosis rate of cells （P<0.05）. Compared with the model group， calycosin （20 μmol·L-1） group displayed high proliferation activity and low apoptosis rate of cells （P<0.05）. Compared with calycosin （20 μmol·L-1） group， both phosphatidylinosirtol-3-kinases （PI3K） inhibitor LY294002 and STAT3 inhibitor Stattic significantly reduced the proliferation activity and increased the apoptosis rate of cells （P<0.05）. The protein expression of p-JAK2， p-STAT3， p-Akt， GP130， and IL-6 in the model group was higher than that in the control group （P<0.05）， and the expression of the above indicators was lower in each treatment group than in the model group （P<0.05）. ConclusionCalycosin can promote the proliferation and inhibit the apoptosis of astrocytes with oxidative injury by inhibiting the phosphorylation of PI3K/Akt pathway and JAK2/STAT3 pathway.
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Objective:To explore the clinical efficacy of dual-kidney transplantation from infant donors to adult recipients.Methods:From December 2012 to November 2020 in Organ Transplant Center First Affiliated Hospital Sun Yat-sen University, rertrospective reviews were conducted for clinical data of 25 pairs of infant donors and adult recipients. The survival rates were calculated for both recipients and transplanted kidneys at Year 1/3/5 post-transplantation. And the postoperative recovery status and the postoperative incidence of adverse events of recipients were observed.Results:The survival rates of recipients were all 95.8% at Year 1/3/5 and those of transplanted kidney and dealth-cancelling transplanted kidney all 87.2%. One case died due to acute inferior-wall cardiac infarction while three others lost renal functions for vascular thrombosis, ureteral stenosis and urinary fistula. Except for loss of renal function and death, the postoperative estimated golmerular fitration rate was (99.35±21.78), (103.11±29.20) and (114.99±28.55) ml/(min·1.73 m 2) at Year 1/2/3 respectively. Conclusions:Selecting proper recipients, standardizing donor acquisition and surgical procedures and strengthening perioperative managements may expand the donor pool. The overall outcomes are excellent for adult recipients with dual-kidney transplantation from donations after infants' death.
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Objective:To summarize the patient profiles and therapeutic efficacies of ABO-incompatible living-related kidney transplantations at 19 domestic transplant centers and provide rationales for clinical application of ABOi-KT.Methods:Clinical cases of ABO-incompatible/compatible kidney transplantation (ABOi-KT/ABOc-KT) from December 2006 to December 2009 were collected. Then, statistical analyses were conducted from the aspects of tissue matching, perioperative managements, complications and survival rates of renal allograft or recipients.Results:Clinical data of 342 ABOi-KT and 779 ABOc-KT indicated that (1) no inter-group differences existed in age, body mass index (BMI), donor-recipient relationship or waiting time of pre-operative dialysis; (2) ABO blood type: blood type O recipients had the longest waiting list and transplantations from blood type A to blood type O accounted for the largest proportion; (3) HLA matching: no statistical significance existed in mismatch rate or positive rate of PRA I/II between two types of surgery; (4) CD20 should be properly used on the basis of different phrases; (5) hemorrhage was a common complication during an early postoperative period and microthrombosis appeared later; (6) no difference existed in postoperative incidence of complications or survival rate of renal allograft and recipients at 1/3/5/10 years between ABOi-KT and ABOc-KT. The acute rejection rate and serum creatinine levels of ABOi-KT recipients were comparable to those of ABOc-KT recipients within 1 year.Conclusions:ABOi-KT is both safe and effective so that it may be applied at all transplant centers as needed.
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Objective:To explore the diagnosis and treatment of focal segmental glomerulosclerosis (FSGS) post-kidney transplantation in children.Methods:Clinical data were retrospectively analyzed for 6 FSGS children after transplantation from 2015 to 2019. Massive proteinuria (3.2-13 g/24 h) occurred at 4 days-49 days post-transplantation. For proteinuria, glucocorticoid plus therapeutic plasma exchange and/or rituximab were provided with supplemental ACEI/ARB drugs. Five cases received tacrolimus as maintenance therapy while another case had cyclosporin A as an initial intensive therapy and switched to tacrolimus.Results:Four cases achieved complete remission after therapy. One recipient showed partial remission. During a follow up period of 11 months to 4 years, serum creatinine remained normal and stable in five cases while one died from severe pulmonary infection.Conclusions:Once FSGS occurs post-transplantation, prompt treatment of pulse glucocorticoid plus therapeutic plasma exchange and/or rituximab with supplemental ACEI/ARB drugs may yield favorable outcomes.
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Objective:To explore the clinical characteristics and outcomes of pediatric kidney transplantations at a single center and discuss the related clinical issues.Methods:From January 1990 to October 2019, clinical data were analyzed retrospectively for 244 pediatric renal transplants. The youngest recipient was aged 1.8 years and the median age of pediatric recipients was 12.2 years. The major disease was primary or hereditary glomerulonephritis ( n=160, 69.0%), congenital anomalies of kidney and urinary tract (CAKUT), cystic renopathy and other hereditary nephropathies ( n=55, 23.7%). The donor sources included traditional deceased donor ( n=42, 17.2%), living-related donor ( n=19, 7.8%) and organ donation ( n=183, 75.0%). The median age of donors was 2 years (0-51) and the median weight 12.0(2.7-72.0) kg. From January 2013 to October 2019, 170 cases), the major induction immunosuppression regimen was anti-thymocyte globulin (ATG) ( n=110, 64.7%) or basiliximab ( n=58, 34.1%). The maintenance regimen was tacrolimus + mycophenolic acid (MPA) + glucocorticosteroids. Finally the outcomes and the complications were analyzed. Results:The survival rates of 244 kidney allograft recipients were 98.1%, 94.5% and 93.4% and the graft survival rates 92.6%, 84.2% and 82.0% at 1/3/5 years respectively. Ten recipients died of accident ( n=2, 20.0%), pneumonia after transplantation ( n=2, 20.0%) and intracranial hemorrhage ( n=2, 20.0%). Thirty-three recipients lost their allografts mainly due to intravascular thrombosis in graft ( n=5, 14.3%), acute rejection ( n=5, 14.3%) and death ( n=9, 25.7%). Besides, among 109 deceased donor allograft recipients, the postoperative outcomes were delayed graft function recovery (DGF) ( n=27, 24.8%), arterial thrombosis ( n=6, 5.5%), venous thrombosis ( n=1, 0.9%), graft perirenal hematoma ( n=6, 5.5%), raft artery stenosis ( n=10, 9.2%) and graft ureteral fistula ( n=1, 0.9%). The incidence of acute rejection was 17.5% and 23.2% at 1/3 year respectively. The recurrent rate of primary disease was 6.9%, including primary FSGS ( n=3, 42.9%) and IgA nephropathy ( n=2, 28.6%). At 1/3 year post-operation, the incidence of pulmonary infection was 16.9% and 22.4% and the incidence of urinary tract infection 26.9% and 31.7%. Excluding recipients with graft failure, the estimated glomerular filtration rate (eGFR) at 1/2/3 year postoperatively was (80.3±25.2), (81.4±27.8) and (71.8±27.6) ml/(min·1.73 m 2)respectively. Conclusions:The outcomes of pediatric renal transplantations are excellent at our center. Future efforts shall be devoted to optimizing the strategies of donor kidney selection and strengthening preoperative evaluations, perioperative and postoperative managements for improving the long-term outcomes of pediatric renal transplantations.
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Objective To investigate the high-fat diet effect on morphology and stiffness of endothelial cells. Methods SD rats were randomly divided into high-fat diet group (AS group, n=3) and control group (CON group, n=3). Rat aortic endothelial cells were obtained from rat thoracic aorta by explant method. Cell morphology was observed under inverted microscopy. The mean fluorescent intensity of F-actin in two groups was calculated by immunofluorescence staining. Cell stiffness was measured by atomic force microscopy (AFM). Results The endothelial cells migrated from tissue plant on the 7th day and formed confluence after cultivation for 14 days. Endothelial cells were identified by factor Ⅷ immunofluorescence staining. Cells in AS group showed enhanced perimeter (P＜0.01), aspect ratio (P＜0.01), and area (P＞0.05), while less circularity (P＜0.01) compared with the cells in control group. The mean fluorescence intensity of F-actin in AS group was significantly higher than that in control group (P＜0.01). AFM showed that the cell stiffness of AS group was significantly higher than that of control group (P＜0.01). Conclusions High-fat diet would change the morphology and stiffness of endothelial cells, which might subsequently affect their normal function and become an important incentive to AS.
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Proprotein convertase subtilisin/kexin type 9 (PCSK9) modulators may attenuate PCSK9-induced low-density lipoprotein receptor (LDLR) degradation in lysosome and promote the clearance of circulating low-density lipoprotein cholesterol (LDL-C). A novel series of tetrahydroprotoberberine derivatives (THPBs) were designed, synthesized, and evaluated as PCSK9 modulators for the treatment of hyperlipidemia. Among them, eight compounds exhibited excellent activities in downregulating hepatic PCSK9 expression better than berberine in HepG2 cells. In addition, five compounds , , , ()-, and ()- showed better performance in the low-density lipoprotein, labeled with 1,1'-dioctadecyl-3,3,3',3'-tetramethyl-indocarbocyanine perchlorate (DiI-LDL) uptake assay, compared with berberine at the same concentration. Compound , selected for evaluation, demonstrated significant reductions of total cholesterol (TC) and LDL-C in hyperlipidemic hamsters with a good pharmacokinetic profile. Further exploring of the lipid-lowering mechanism showed that compound promoted hepatic LDLR expression in a dose-dependent manner in HepG2 cells. Additional results of human related gene (hERG) inhibition assay indicated the potential druggability for compound , which is a promising lead compound for the development of PCSK9 modulator for the treatment of hyperlipidemia.
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Objective To assess the efficacy and safety of febuxostat in the treatment of hypemricemia in renal transplant recipients.Methods A total of 124 renal transplant patients with hyperuricemia receiving febuxostat between June 2016 and July 2018 were retrospectively analyzed.Uric acid (UA),liver function and renal function parameters before and 3 months after treatment were compared.Adverse events,recipient and renal allograft survival were recorded throughout the follow-up period.Results Serum level of uric acid significantly decreased after 3-month treatment (P＜0.001).And 66.1％ of them achieved target UA level at Month 3 after dosing.Estimated glomerular filtration rate (eGFR) was maintained.No severe adverse event was observed.All recipient and renal grafts survived during the follow-up period.Conclusions Febuxostat is both effective and safe in the treatment of hyperuricemia in renal transplant.
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Objective To explore the clinical outcome of renal transplantation and analyze the risk factors influencing the kidney allograft survival after transplantation.Methods The clinical data of 524 cases of renal transplantation between January 2007 and December 2015 were retrospectively analyzed.Serum creatinine was determined,and glomerular filtration rate(GFR) was estimated.The 1-,2-and 3-year patient and graft survival after transplantation was calculated.Adverse events were recorded.Results The median follow-up time was 17.2 months.The 1-,2-and 3-year graft survival rate after transplantation was 97％,95.8％ and 95.3％,respectively.The 1-,2-and 3-year patient survival rate after transplantation was 97.8％,97％ and 97％,respectively.The eGFR was (67.6 ± 24.1),(68.9±24.2) and (72.7 ± 26.2) ml·min-1 ·1.73 m-2 at 1st,2nd and 3rd year after transplantation.The incidence of delayed graft function(DGF) was 20.6％ (108/524).Multivariate analysis revealed donor type (P =0.005) and the terminal creatinine (P＜0.001) were the independent risk factors of DGF.Elder recipients (P =0.004),recipients with diabetes(P =0.031),preoperative positivity of panel reactive antibody(PRA) (P =0.023),and donor with hypertension (P =0.046) were risk factors influencing the kidney allograft survival.Conclusion Kidney transplantation showed good outcomes at 3rd year after transplantation.The recipient age,recipient's history of diabetes,preoperative PRA and donor's history of hypertension are independent risk factors for renal graft survival.
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Objective To screen and verify the proteins interacting with phosphorylation cluster of DNA dependent protein kinase catalytic subunit((DNA-PKcs) by yeast two-hybrid assay.Methods To know the proteins interacting with DNA-PKcs phosphorylation cluster,yeast two-hybrid assay was applied to screen the cDNA library of human hepatic tissue with a previously constructed plasmid pGBKT7-DPC.The positive clones were further identified by PCR,rotary validation and sequence analysis.Then the eukaryotic expression vectors of the bait protein and screened positive clone proteins were constructed and transfected into human embryonic kidney 293T cells to detect whether the proteins could been expressed correctly.At last,the bait protein and screened positive clone proteins were co-transfected into 293T cells and protein interaction was detected with Co-Immunoprecipitation (Co-IP) assay.Results After two rounds of screening using the yeast two-hybrid assay,12 candidate clones were obtained.Then 7 clones with different insert fragments were identified by PCR,and 3 positive proteins interacted with DNA-PKcs phosphorylation cluster were further verified by rotary validation.Sequencing analysis demonstrated that these 3 proteins were MBNL1,SIK2 and YY1AP1,respectively.Accordingly,the eukaryotic expression vectors of bait protein and 3 positive clone proteins were constructed successfully and expressed correctly in 293T ceils.Finally,the Co-IP assay confirmed that these 3 positive clone proteins could interact with DNA-PKcs phosphorylation cluster.Conclusions Proteins interacting with DNA-PKcs phosphorylation cluster are successfully screened and identified.
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Objective To explore the effects of co‐cultured heat‐treated candida glabrata with rat tracheal epithelial (RTE) cells on the expression of Dectin‐1 and the production of IL‐6 and TNF‐α.Methods RTE cells in vitro were co‐cultured with heat‐treated candida glabrata bacteria liquid for 2 ,4 ,6 h ,while without co‐cultured RTE cells were used as control group .We observed the morphological changes of RTE cells ,detected the protein expressions of Dectin‐1 by Western blot ,used real‐time PCR to detecte the mRNA expressions of IL‐6 and TNF‐αand measured protein expression of IL‐6 and TNF‐αby ELISA .Results With the pass‐ing of time ,the RTE cells were damaged extensively and the expression of Dectin‐1 ,IL‐6 and TNF‐αbecame more and more signifi‐cant .Obviously ,there had significant difference in the expression of Dectin‐1 ,IL‐6 and TNF‐α between the co‐cultured 2 h group and the control group ,the co‐cultured 4 h group and the co‐cultured 2 h group ,the co‐cultured 6 h group and the co‐cultured 4 h group (P<0 .05) .Conclusion RTE cells have natural immune function .The Dectin‐1 involves in the recognition of heat‐treated Candida glabrata ,activating secretion of IL‐6 and TNF‐αand mediating inflammatory reaction .IL‐6 plays a negative regulation role .
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Objective To study the roles of Toll-like receptor 2(TLR2)and Toll-like receptor 4(TLR4)in the progress of pulmo-nary infection with candida glabrata in experimental mice .Methods The mice were divided into three groups ,the control group (group A) ,mice infected with candida glabrata(group B)and immunosuppressive mice infected with candida glabrata(group C) .6 mice in each group were killed at 1st and 3rd day after the success in pulmonary infection with candida glabrata .The lung tissues from each group were collected for pathological analysis and PT-PCR to detect the expression level of TLR2 and TLR4 .Protein TNF alpha(TNF-α)level was measured by ELISA method .Results The pathological analysis showed the structure was normal and there was no inflammatory reaction in lungs in group A .The infiltration of inflammatory cells ,weak injuries but no conidia or hyphae in lungs were found in group B .Most of the alveolar collapse and severe damage ,part of the expansion ,a large number of in-flammatory cells infiltration ,accompanying with conidia and hyphae accumulation were observed in group C .The expression levels of TLR2 mRNA and TNF-αprotein in mice lungs at the 1st day and the 3rd day in group C were significantly higher than in group A and B(P<0 .05) ,while group B was higher than group A(P<0 .05);The expression levels of TLR2 ,TLR4 mRNA and TNF-αprotein in group C at the 3rd day was higher than the 1st day(P<0 .01) ,meanwhile ,the expression of TLR4 was higher than in group A(P<0 .05) .Conclusion TNF-αand TLR2 expression maybe involve in the infection and process of candida glabrata ,and TLR4 may play a synergistic effect .
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Objective To explore the significance of the Self -assessments of Voice Handicap Index (VHI) and The objective detection of Dysphonia Severity Index (DSI) in polyp of vocal cord perioperatively .Methods The Self -assessments of VHI had been completed in 28 patients with polyp of vocal cord before the operation and one week after the operation ,including function(F) ,physiological(P) ,emotion(E) ,and the sum denoted by T .In the meantime ,The DSI had been completed in these patients by acoustic analysis of DIVAS 2 .5 .After one month ,19 pa-tients had been re -examined .Results The DSI value was increasing ,but there was no significant difference be-tween pre-operation and one week after the operation(P>0 .05) ,indicating that the function of vocal cord was not recovered .In VHI scale ,the parameters were significantly different between the time preoperatively and one week postoperatively in F ,E and T(P0 .05) ,indicating that the function of vocal cord was not completely recovered as same as the result of DSI .After one month ,there were significant differences in DSI and VHI ,which showed a full recovery of vocal cord fanction .Conclusion The Self -assessments of VHI and The objective detection of DSI in polyp of vocal cord can evaluate simultaneouly the function of laryngeal .Additionally ,there was a good correlation between the DSI and the VHI score .