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Objective:Clinical research is widely carried out in medical institutions, and there are differences in the use of research and conventional medical funds. This paper aimed to analyze the compliance issues of insurance fund in clinical research and explore the management strategies in the institutions conducting clinical trials or research.Methods:By consulting the literature, questionnaire, and work practice, this paper analyzed the current situation and existing problems of the compliance of the medical insurance fund in domestic clinical trials, proposed targeted management measures for the use of funds, and standardizes the corresponding workflow.Results:This paper summarized three payment methods of research-related funds and analyzed the main problems at present, including the definition of trial requirements, the payment of combined drugs specified in the protocol, the particularity of medical device trials, the payment of adverse events in clinical trial, the insurance of post-marketing research and clinical trial. According to the regulatory requirements and work practices, the corresponding management countermeasures were sorted out, including that the project funds and insurance audit should be carried out inside the medical institution. Clinical research, medical insurance management departments, ethics committees, and other departments should collaborate on establishing and improving a compliance management system. The research team should strengthen the management of adverse events, strengthen the publicity and education of the participants, and make good use of the sharing platform.Conclusions:The research institution should establish standardized and feasible processes, the research team should strengthen the management, and the use of the sharing platform is conducive to ensuring the compliance of the medical insurance fund and protecting the interests of the participants.
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Objective:To observe and preliminarily explore the effect of mogroside on oxidative stress of retinal pigment epitheliaum (RPE) cells induced by hydrogen peroxide (H 2O 2) and its possible mechanism. Methods:A experimental study. The RPE cells were divided into control group, H 2O 2 group, silent information regulator of transcription 1 (SIRT1) inhibitor EX527 group (EX527 group), mogroside group, mogroside+EX527 group. Methyl thiazolete trazolium method was used to detect cell survival rate. Flow cytometry was used to detect cell apoptosis rate. 2' ,7'-dichlorodihydrofluorescein diacetate fluorescent probe method, xanthine method and enzyme-linked immunosorbent assay method were used to detect the level of reactive oxygen species (ROS), superoxide dismutase (SOD) activity and malondialdehyde (MDA) content in cells respectively. Real-time quantitative polymerase chain reaction and Western blot were used to detect relative expressions of SIRT1, nuclear factor erythroid-2-related actor 2 (Nrf2), heme oxygenase-1 (HO-1) mRNA and protein in cells. One-way ANOVA was used for comparison among groups. The pairwise comparison between groups was tested by the least significant difference t test. Results:Compared with the control group, the H 2O 2 group cell survival rate decreased, the apoptosis rate increased, the ROS level in the cells increased, the SOD activity decreased, the MDA content increased, and the relative expression of SIRT1, Nrf2, HO-1 mRNA and protein decreased ( P<0.05). Compared with H 2O 2 group, the cell survival rate decreased, apoptosis rate increased, the cell ROS level increased, SOD activity decreased, MDA content increased, SIRT1, Nrf2, HO-1 mRNA and protein expression decreased in EX527 group ( P<0.05); the cell survival rate increased, apoptosis rate decreased, ROS level decreased, SOD activity increased, MDA content decreased, and the relative expression of SIRT1, Nrf2, HO-1 mRNA and protein increased in mogroside group ( P<0.05). Compared with the mogrosides group, the cell survival rate decreased, the apoptosis rate increased, the level of ROS increased, SOD activity decreased, MDA content increased, SIRT1, Nrf2, HO-1 mRNA and protein decreased in mogrosides+EX527 group ( P<0.05). Conclusions:Mogrosides can alleviate the oxidative stress response of visual RPE cells induced by H 2O 2, promote cell proliferation, and reduce cell apoptosis. Mogrosides may exert antioxidant effects by activating the SIRT1/Nrf2 signaling pathway.
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Objective:To evaluate the efficacy and safety of non-osteotomy total hip arthroplasty (THA) in the treatment of Crowe IV developmental dysplasia of the hip (DDH).Methods:From Jan 2013 to Sep 2021, 46 patients (46 hips) in our department who underwent total hip arthroplasty without osteotomy for unilateral Crowe IV DDH were retrospectively analyzed, including 6 males and 40 females, with an average age of 41.2±7.2 years (25-61 years). The reduction of the femoral head was achieved successfully through moderate upward-posterior displacement of the movement center, gradual osteotomy of the calcar femorale, proper sinking of the femoral prosthesis and sufficient soft tissue release. The evaluation indexes included the basic condition of the operation (operation time, blood loss, blood transfusion, volume), clinical evaluation (Harris score of hip joint function, patient satisfaction, Trendelenburg sign), imaging evaluation (measurement of limb length and pelvic inclination) and incidence of complications.Results:The mean follow-up time was 72.2±8.8 months (8-101 months). The operation time was 97.2±12 min (84-112 min). The average intraoperative bleeding volume was 550±60 ml (350-850 ml). No patient had periprosthetic infection or fracture, no periprosthetic osteolysis or prosthesis loosening, and no patient needed revision surgery at the last follow-up of all cases. The average HHS score of patients increased from 42.5±12.3 points before surgery to 89.2±10.8 points at the last follow-up, and the difference was statistically significant ( t=19.35, P<0.001). Patient self-rated satisfaction: none was very dissatisfied, 1 patient were less satisfied (2%, 1/46), 4 patients were average (9%, 4/46), 19 patients were relatively satisfied (41%, 19/46), and 22 patients were very satisfied (48%, 22/46). The Trendelenburg sign of 46 cases was positive before operation, and all were negative at the last follow-up. The patients' true leg length discrepancy (LLD) measurement was -2.5±0.6 mm before surgery and 11.5±3.2 mm at the last follow-up ( t=29.17, P<0.05). Patients' perceived LLD was 28.2±5.1 mm before surgery and 3.4±1.4 mm at the last follow-up ( t=32.18, P<0.05). The length of the residual calcar femorale was 3.2±0.4 mm after THA. The limb extended distance of affected limb was 45.2±4.6 mm. The preoperative iliolumbar angle was -6.5°±2.3°, which returned to -0.5°±1.3° at the last follow-up ( F=651.97, P<0.05). Conclusion:For patients with unilateral type IV DDH, non-osteotomy THA is a safe and effective surgical method with simple operation and few complications. It can quickly correct pelvic tilt and lumbar compensatory scoliosis postoperatively.
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Objective:To investigate the safety and efficacy of a combined anterior and posterior approach in total hip arthroplasty (THA) for fused/ankylosed hip.Methods:37 patients who underwent THA for fused/ankylosed hip from January 2015 to December 2020 were retrospectively analyzed, including 28 males and 9 females, with an average age of 47.9±12.0 years (range, 26-72 years). Etiologies included 23 cases of ankylosing spondylitis, 9 cases of infectious arthritis of the hip in youth, and 5 cases of traumatic arthritis after acetabulum or femoral neck fracture. All patients underwent THA with combined anterior and posterior approach. These factors include operation time, blood loss, amount of transfused blood, blood transfusion rate, preoperative and postoperative Harris score, postoperative range of motion of the hip, and perioperative complications, etc. were evaluated. Postoperative radiography of the hip was performed to evaluate acetabular abduction angle, anterior inclination angle, the prosthesis fixation, osteolysis and heterotopic ossification around the hip.Results:A total of 37 patients were enrolled. The mean operative time was 147.6±16.8 min (range, 129-190 min); the mean estimated blood loss (EBL) was 850.0±10.8 ml (range, 600-1,200 ml); the blood transfusion rate was 59% (22/37), and the mean blood transfusion was 420±45.0 ml (range, 0-800 ml). All patients were followed up for 4.2±0.9 years (range, 1.2-7.2 years). The average abductor angle of the acetabular was 43.7°±5.4° (range, 31°-55°), and the average inclination angle was 20.9°±6.7° (range, 10°-35°); the preoperative Harris score was 47.1±9.9 (range, 40-55) and the mean Harris score at the last follow-up was 83.4±12.4 (range, 75-90). The preoperative range of motion of the hip in all directions was 0°. Postoperative hip range of motion was good, with a mean hip flexion of 95.5°±12.2° (range, 80°-110°), mean extension of 10.5°±3.4° (range, -10°-25°), and mean abduction of 38.0°±8.2° (range, 10°-50°). Postoperative complications were minor, including 2 case with poor wound healing, 2 cases with paresthesia or tingling sensation in the anterior or anterolateral thigh, which returned to normal within 3 months after surgery, and no deep infection or dislocation occurred. The acetabular cup was in poor position in 2 cases and the femoral stem was varus in 1 case, but the prosthesis was stable and no treatment was needed. All the acetabular cups and femur stems were confirmed with bone ingrowth on the last follow-up radiographs, and one patient had heterotopic ossification (Brooker grade 1). No osteolysis or wear of the acetabular liner was observed.Conclusion:Combined anterior and posterior approach (Gibson posterolateral approach + modified Hardinge approach) in THA for fusion/ankylosed hip can fully expose the operative field and sufficiently release the soft tissue, and the function of hip recovered well postopratively.
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Objective:To investigate effect and underlying lipid-lowering mechanisms of catalpol in non-alcoholic fatty liver disease(NAFLD).Methods:In vivo model of NAFLD was established with high-fat diet-fed ICR mice for 8 weeks. Low(50 mg/kg), medium(150 mg/kg), and high(300 mg/kg) doses of catalpol were administered, and the body weight, liver weight, hepatic index, and biochemical parameters of the mice were analyzed. Free fatty acid-induced LO2 in human hepatocytes to establish NAFLD cell model. Quantitative realtime PCR reaction to detect fatty acid synthesis-related gene levels. Western blotting assay was adopted to analyze proteins in the endoplasmic reticulum stress(ERS)-mediated protein kinase RNA-like endoplasmic reticulum kinase(PERK)-eukaryotic translation initiation factor 2α(eIF2α) signaling pathway. Results:Compared with model mice, body weight [(39.43±1.84)g, (34.01±1.83)g, (32.28±1.11)g vs(42.17±1.37)g, all P<0.001], liver weight [(1.03±0.06)g, (0.79±0.05)g, (0.64±0.04)g vs(1.30±0.13)g, P<0.01 or P<0.001], and liver index [(2.60±0.09)%, (2.32±0.09)%, (1.99±0.11)% vs(3.07±0.30)%, P<0.05 or P<0.001] were reduced in low, medium, and high doses of catapol model. Medium and high doses of catalpol diminished total cholesterol, triglyceride, low density lipoprotein-cholesterol, aspartate aminotransferase, and alanine aminotransferase( P<0.01 or P<0.001), increased high density lipoprotein-cholesterol( P<0.01 or P<0.001). In the cell model, elevated levels of both fatty acid synthesis genes and PERK-eIF2α pathway proteins were attenuated by catalase, and this attenuation was reversed by signaling pathway agonists. Conclusion:The Chinese herb catalpol may play a role in improving NALFD by regulating the ERS-mediated PERK-eIF2α signaling pathway.
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Objective:To investigate the ultrasonographic characteristics and misdiagnosis of hepatic pregnancy.Methods:The clinical data and preoperative ultrasound characteristics of 2 cases of hepatic pregnancy admitted to the Affiliated Hospital of Jining Medical College were retrospectively summarized, and related literatures were retrieved from January 1, 1956 to December 31, 2021.Results:A total of 37 cases of hepatic pregnancy were included, among which 35 cases showed liver nodules on ultrasound examination, which were classified into four types: pregnancy cystic (24 cases), thick-walled cystic echo (6 cases), heterogeneous hyperecho (3 cases) and mixed cyst solid echo (2 cases), the exact diagnosis coincidence rate of each type was 100%, 4/6, 1/3, 0, respectively. Among the 35 liver nodules, 29(82.8%) cases were diagnosed correctly by ultrasound, 1 case was misdiagnosed as choriocarcinoma liver metastasis, 1 case was misdiagnosed as hepatic hemangioma, and 4 cases were diagnosed as liver nodules without suggestive diagnosis. No liver nodules were found in the other 2 cases, including 1 case misdiagnosed as retroperitoneal hematoma and 1 case misdiagnosed as ruptured ovarian cyst.Conclusions:Ultrasonography of hepatic pregnancy may show pregnancy sac, thick-walled cystic echo, hyperecho and mixed echo in liver. The first type is easy to be diagnosed accurately by ultrasound, while the other types are easy to be misdiagnosed. Combined with clinical conditions and the result of enhanced computed tomography (CT), the correct diagnosis rate of hepatic pregnancy can be improved.
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Objective:To investigate the imaging features of iris fluorescein angiography (IFA) combined with fluorescein fundus angiography (FFA) in diabetic iridopathy.Methods:A cross-sectional study was conducted.Sixty-five eyes of 44 patients with proliferative diabetic retinopathy (PDR) combined with diabetic iridopathy (DI) were enrolled in Henan Eye Hospital from May 2013 to May 2020.Patients were divided into non-proliferative diabetic iridopathy (NPDI) group and rubeosis iridis group according to the imaging results.Ophthalmic examinations including visual acuity, intraocular pressure, slit lamp miacroscopy, IFA and FFA were carried out in all patients.IFA was used to detect the iris imaging characteristics and the regression time of fluorescein in anterior chamber, and FFA was used to observe the retinal image characteristics and the incidence of optic disc neovascularization.To avoid the statistical error of recording the IFA examination time of the contralateral eye, only the relevant data of the affected eyes were analyzed.This study adhered to the Declaration of Helsinki.The study protocol was approved by the Ethics Committee of Henan Eye Hospital (No.HNEECKY-2020[06]). Written informed consent was obatined from all patients before any medical examination.Results:Among the patients, 30 cases (50 eyes) were with NPDI and 14 cases (15 eyes) were with rubeosis iridis.The fluorescein regression time in anterior chamber of NPDI group was (3.37±0.11) minutes, which was significantly shorter than (6.02±0.29) minutes of rubeosis iridis group ( t=8.541, P<0.001). Strong fluorescence of retinal neovascularization was observed in both groups.The incidence of optic disc neovascularization in NPDI group was 20% (6/30), which was significantly lower than 50% (7/14) in rubeosis iridis group ( P=0.04). Conclusions:Diabetic rubeosis iridis can be diagnosed by the imaging features of IFA and the fluorescein regression time in anterior chamber.PDR combined with optic disc neovascularization should be evaluated by FFA combined with IFA.
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Proliferative vitreoretinopathy (PVR) is a common complication of retinal detachment surgery, also an important blinding cause.Besides retinal pigment epithelial cells, retinal glial cells also are mainly involved in the formation and development of PVR.Retinal glial cells include Müller cells, astrocytes and microglia.The glial cells not only can be activated to change the morphology of cells, but also secrete growth factors and cytokines, synthesize extracellular matrix, and participate in the formation of PVR.The factors and synthetic extracellular matrix secreted by retinal glial cells promote the proliferation of retinal glial cells, which accelerates the development of PVR.This article described the interaction between the activation of retinal glial cells, astrocytes, growth factors secreted, cytokines secreted, extracellular matrix synthesized by glial cells and PVR formation, and the effect of non-coding RNAs on glial cells and PVR.
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Objective:To observe the incidence of diabetic iridopathy and optic disc neovascularization in the contralateral eyes of proliferative diabetic retinopathy (PDR) with proliferative diabetic iridopathy (PDI).Methods:A retrospective case-control study. From February 2014 to May 2020, 72 eyes of 36 patients with PDR and PDI who underwent iris fluorescein angiography (IFA) combined with fluorescein fundus angiography(FFA) at the Henan Eye Institute were enrolled in the study. Among them, there were 34 eyes in 17 males and 38 eyes in 19 females; the average age was 62.3±4.7 years. All patients underwent best corrected visual acuity(BCVA), intraocular pressure, IFA combined with FFA examination. The BCVA examination was performed using the international standard visual acuity chart, which was converted into logarithm of the minimum angle of resolution BCVA for statistic analysis. According to PDI staging, patients were divided into early rubeosis iridis(RI) group and neovascular glaucoma (NVG) group, with 28 and 8 cases respectively. Compared with NVG group, RI group had better BCVA and intraocular pressure, and the difference was statistically significant( t=6.433, 10.619; P=0.000, 0.011). The incidence of PDI and the incidence of binocular optic disc neovascularization in the two groups were compared, and Fisher's exact probability method was used for comparison. Results:The results of the IFA examination showed that in the RI group, the pupil border and the iris surface of the iris of the affected eyes had strong neovascular bud-like or patchy fluorescence; the pupil border and the middle of the iris of the patients in the NVG stage had strong neovascular cluster-like fluorescence. Among the contralateral eyes in the RI group, 6 eyes (21.4%, 6/28) were with PDI (stage RI), 21 eyes (75.0%, 21/28) were with non-PDI (NPDI), and 1 eye (1/ 28, 3.6%) were absence of diabetic iris disease. Among the contralateral eyes in the NVG group, there were 5 eyes with PDI (62.5%, 5/8), including 4 eyes with RI stage, 1 eye with NVG stage (12.5%, 1/8); 3 eyes with NPDI. The image of IFA in patients with NPDI early showed as punctate fluorescence in the local small blood vessels of the iris itself. The incidence of PDI in the contralateral eye of the RI group was lower than that of the NVG group, and the difference was statistically significant ( P=0.040). The results of FFA examination showed that 9 (32.1%, 9/28) and 8 (28.6%, 8/28) eyes of the affected eye and the contralateral eye in the RI group were combined with optic disc neovascularization. In NVG group, there were seperately 6 eyes (75.0%, 6/8) in the affected eyes and the contralateral eyes with optic disc neovascularization. The differences in the incidence of optic disc neovascularization between the two groups of the affected eyes and the contralateral eye were statistically significant ( P=0.046, 0.040). Conclusion:The incidence of PDI and optic disc neovascularization in the contralateral eye of PDR and PDI, RI is lower than that of NVG.
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Objective:This study aimed at analyzing risk factors associated with surgical outcomes of neonatal total anomalous pulmonary venous connection (TAPVC) in our center.Methods:A total of 105 neonates who underwent surgical repair for TAPVC from January 1st, 2009 to January 1st, 2018 were retrospectively analyzed. The anatomical types of TAPVC included supracardiac 42(40%, 42/105), cardiac 21(20%, 21/105), infracardiac 36(34.3%, 36/105), and mixed 6(5.7%, 6/105). The Cox proportional hazards analysis was used to analyze the risk factors related to postoperative pulmonary venous obstruction (PVO) and mortality. Kaplan- Meier analysis was used to analyze the overall survival rates. Results:Twenty-six patients (24.8%, 26/105) were diagnosed with preoperative PVO. The 30-day, 1 year, and 5 years survival rate was 92.4%, 86.7%, and 86.7% respectively. Postoperative PVO occurred in 17 patients (16.2%, 17/105). Preoperative acidosis, low surgical weight, prolonged duration of cardiopulmonary bypass time, increasing postoperative central venous pressure (CVP), and reoperation were risk factors associated with mortality. Preoperative acidosis ( P<0.001), prolonged duration of cardiopulmonary bypass time ( P<0.001), and increasing postoperative CVP ( P=0.005) were independent risk factors for mortality. Mixed TAPVC, preoperative acidosis, low surgical age, prolonged cardiopulmonary bypass time, postoperative pulmonary arterial hypertension were risk factors associated with postoperative PVO. Prolonged cardiopulmonary bypass time ( P=0.029), postoperative pulmonary arterial hypertension ( P<0.001), and mixed TAPVC ( P=0.017) were independent risk factors associated with postoperative PVO. Conclusion:The surgical outcomes of neonatal TAPVC in our center were acceptable, with low mortality rate and incidence of PVO. However, neonates with preoperative acidosis, prolonged duration of cardiopulmonary bypass time, and increased postoperative CVP had a poor prognosis. Patients with mixed TAPVC were at increased risk for postoperative PVO.
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Objective:To analyze the imaging characteristics of invasive fibromatosis from breast parenchyma, and to explore the clinical value of multimodal ultrasound.Methods:The two-dimensional, color Doppler and elastic imaging sonographic manifestations and pathological features of 12 cases of breast invasive fibromatosis included in the Affiliated Hospital of Jining Medical College from October 2015 to October 2020 were studied retrospectively.Results:Two dimensional ultrasound showed that 12 cases of breast invasive fibromatosis grew in parallel, with different sizes, unclear boundary and no complete capsule. The edge morphology of 7 cases (7/12) showed crab foot like changes; The tumor showed solid heterogeneous hypoechoic, punctate hyperechoic in 3 cases (3/12), and echo attenuation behind the tumor in 3 cases (3/12); The blood flow in the tumor was mainly grade 1-2 (9/12). Twelve tumors were examined quantitatively by virtual touch tissue quantifification. The shear wave velocity was (3.08±0.75)m/s. The diagnostic accuracy of multimodal ultrasound in invasive fibromatosis of breast (10/12) was significantly higher than that of conventional ultrasound (3/12, χ 2=8.224, P=0.004). The gross manifestation of the tumor: the section was gray or gray yellow, with unclear boundary and no capsule; Microscopic findings: fascicular fibroblasts and myofibroblasts proliferated, accompanied by varying degrees of glassy degeneration, acellular atypia, nuclear division and necrosis. Conclusions:Conventional ultrasound is difficult to distinguish invasive fibromatosis and malignant tumor of breast, and its ultrasonic manifestations are closely related to pathological features. The combined application of multimodal ultrasound can significantly improve the diagnostic coincidence rate of the disease and has high clinical application value.
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Objective:To establish and apply the intervention model of professional attitude for nursing undergraduates in universities of Traditional Chinese Medicine (TCM), and to explore its effect on the professional attitude of the nursing undergraduates.Methods:The study started in August 2018 included two steps. The first step was to develop intervention model of professional attitude for nursing undergraduates in universities of TCM based on the three-stage theory of Kelman ′s attitude formation and with Delphi method. After two rounds of expert consultation, the intervention model of professional attitude for nursing undergraduates in universities of TCM was finally formed, including three intervention stages, a total of 41 intervention measurements. The second step was to use the historical control research method. Grade 2016 nursing undergraduate (the year 2016—2020) in Beijing University of Chinese Medicine was taken as the experimental group ( n=86) and adopted the intervention model. Grade 2012 nursing undergraduate (the year 2012—2016) in the same university was taken as the control group ( n=56) and did not adopted the intervention model during their university study. The two groups were continuously evaluated by "Scale for Nurse ′s Professional Attitude" during their four years in the university. The measurement time was within one week of admission and the 16th week of the second semester of each academic year, with a total of 5 times of measurement. Results:Repeated measurement design showed that the scores of Scale for Nurse′s Professional Attitude of the two groups changed with time. At the end of the first year, the score of the scale in the experimental group was (6.06±0.95) points, higher than (5.41±1.08) points of the control group ( t value was 3.765, P<0.001). There was no difference between the two groups at other time points ( P>0.05). Conclusions:The intervention model of professional attitude for nursing undergraduates in universities of TCM has good operability and can improve the professional attitude of nursing undergraduates in universities of TCM during their freshman year. The intervention model needs to be further adjusted to improve the professional attitude in the subsequent learning stage.
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@#Objective To sum up the experience of the primary modified single-patch (MSP) technique applied in our hospital for children with complete atrioventricular septal defect (CAVSD). Methods The clinical data of 141 children who underwent primary MSP technique for CAVSD between June 2009 and December 2017 were retrospectively analyzed, including 62 males and 79 females with a median age of 6 (3, 11) months and a median weight of 5.8 (4.5, 7.0) kg. According to Rastelli classification, there were 116 patients in type A, 14 in type B and 11 in type C. Among them, 15 patients were diagnosed with Down’s syndrome. Cardiopulmonary bypass time, aortic cross clamp time, atrioventricular valve regurgitation and other clinical data were recorded during and after operation. Results Postoperatively, 17 patients suffered from severe left atrioventricular valve regurgitation (LAVVR) and 6 patients severe right atrioventricular valve regurgitation (RAVVR). In the follow-up period, 5 patients suffered from severe LAVVR and 1 patient severe RAVVR. Left ventricular outflow tract obstruction (LVOTO) appeared in 1 patient during follow-up period and none at the end of follow-up. There were 5 early deaths and 2 late deaths. Twelve patients underwent reoperation with a median interval time of 268 (8, 1 270) days. Conclusion MSP technique is a wise surgical strategy for CAVSD children with good outcomes, improved postoperative mortality and decreased atrioventricular valve regurgitation.
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Objective:To investigate the effect of microRNA-155(miR-155) on transforming growth factor β2 (TGF-β2)-induced epithelial-mesenchymal transition of human retinal pigment epithelial cells and its mechanism.Methods:The retinal pigment epithelial cell ARPE-19 cell line was used as the research object.The cells cultured with DMEM medium were served as the control group and the cells cultured with DMEM medium containing 10 ng/ml TGF-β2 were served as the TGF-β2 group.The ARPE-19 cells transfected with miR-155 inhibitor were set as the miR-155 inhibitor group and the ARPE-19 cells transfected with miR-155 negative control were set as the miR-155 negative control group, and the cells in the two groups were cultured in DMEM medium containing 10 ng/ml TGF-β2.After 48 hours cell culture, reverse transcription-PCR was used to detect the expression of miR-155 in each group, and scratch migration test and Transwell chamber test were used to detect cell migration and invasion ability, and Western blot was used to detect the expressions of phosphate and tension homology deleted on chromosome ten gene (PTEN), phosphatidylinositol 3-kinase (PI3K), protein kinase B (Akt), p-Akt and epithelial mesenchymal markers E-cadherin (E-cad), zonula occludens protein 1 (ZO-1), F-actin, α-smooth muscle actin (α-SMA), fibronectin 1 (FN-l) vimentin, proteins.The target gene prediction library predicted miR-155 target gene and fluorescein enzyme reporter vectors were used to identify target genes.Results:After 48 hours of culture, the cells in the control group were in good condition with tight adherence and regular shape.The cells in the TGF-β2 group showed more obvious spindle shape with loose arrangement, and most of the cells were fibrous.The relative expression level of miR-155 in the cells of TGF-β2 group was 0.92±0.14, which was significantly higher than 0.35±0.06 of the control group ( t=7.242, P=0.003). The relative expression level of miR-155 in the cells of miR-155 inhibitor group was 0.21±0.03, which was significantly lower than 0.98±0.09 of the miR-155 negative control group ( t=12.421, P<0.01). The migration rate was higher and the number of cells passing through basement membrane was more in the TGF-β2 group than those of the control group, and the migration rate was higher and the number of cells passing through basement membrane of miR-155 was more in the miR-155 negative control group than those of the miR-155 inhibitor group, and the differences were statistically significant (all at P<0.01). Compared with the control group, the relative expression levels of PTEN, E-cad, ZO-1, F-actin protein were decreased, while the relative expression levels of PI3K and the p-Akt/Akt ratio were increased, and the relative expression levels of α-SMA, FN-1, vimentin proteins were increased in the TGF-β2 group, and the differences were statistically significant (all at P<0.01). Compared with the miR-155 negative control group, the relative expression levels of E-cad, ZO-1, F-actin and PTEN proteins were increased, while the relative expression levels of α-SMA, FN-l, vimentin, PI3K and the p-Akt/Akt ratio were decreased in the miR-155 inhibitor group, and the differences were statistically significant (all at P<0.01). Target gene prediction library prediction and luciferase reporter vector identification confirmed that PTEN was a downstream target gene of miR-155. Conclusions:miR-155 can promote the TGF-β2-induced epithelial-mesenchymal transition progress in human retinal pigment epithelial cells, and its mechanism may be related to inhibiting the expression of the target gene PTEN and stimulating the activation of the PI3K/Akt signaling pathway.
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Proliferative vitreoretinopathy (PVR) is an ocular fundus disease involving multiple cytokines.Its important pathological process is epithelial-mesenchymal transition (EMT) of retinal pigment epithelium (RPE) cells.Tumor necrosis factor (TNF) is an important inflammatory response inducing factor, which can be produced by activated RPE cells, microglia, monocytes and macrophages, and then participate in the occurrence and development of PVR.In addition to cytokines, epigenetic factors such as DNA methylation also play an important role in the development of PVR, in which methyl-CpG binding protein 2(MeCP2) is involved in EMT and fibrosis, and is highly expressed in PVR membrane.The positive expression of MeCP2 is also found in transformed RPE cells and microglia.It is speculated that MeCP2 plays an important role in the occurrence and development of PVR.TNF can also stimulate the expression of MeCP2.This article reviews the role of MeCP2 and the interaction between TNF and MeCP2 in the formation of PVR.
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Objective To investigate the role of methyl-CpG-binding protein 2 (MeCP2) in the regulation and epithelial-mesenchymal transition (EMT) of human lens epithelial cells (LECs) and its possible mechanism.Methods Human LEC lines (SRA01/04) were divided into MeCP2-mimic group,MeCP2-NC group and small interferening RNA-MeCP2 (si-MeCP2) group,and MeCP2 analog plasmid,blank plasmid and MeCP2 si-RNA plasmid was used respectively to transfect the cells.The expression of MeCP2 mRNA in the cells was detected by using real-time PCR 24 hours after transfection.At 48 hours after transfection,the migration rate of the cells was evaluated by scratching test,and the expression of Wnt3a protein in the cells was detected by immunofluorescence stainning.The relative expressions of β-catenin,E-cadherin,Vimentin,matrix metallo proteinase (MMP)-9,MMP-7 and secreted frizzled-related protein 5 (SFRP5) proteins in the cells were detected by Western blot.Results After 24 hours of transfection,the relative expression of MeCP2 mRNA in the cells was significantly different among the MeCP2-mimic group,MeCP2-NC group and si-MeCP2 group (F =4 773.00,P＜0.00 1).The migrating rate of the cells in the MeCP2-mimic group,MeCP2-NC group and si-MeCP2 group was (57.45±5.20)％,(32.71± 10.02)％ and (17.77±9.22)％,respectively,showing a significant difference among the three groups (F=124.00,P＜0.001),and the migrating rate of the cells in the si-MeCP2 group was significantly lower than that of the MeCP2-mimic group or MeCP2-NC group (both at P＜0.001).The relative expressing intensity (absorbance) of Wnt3a in the cells of the MeCP2-mimic group,MeCP2-NC group and si-MeCP2 group was 75.92 ± 6.10,52.03 ± 5.22 and 28.75 ± 3.39,respectively,with a significant difference among three the groups (F=221.30,P＜0.001),and the relative expressing intensity (absorbance) of Wnt3a in the cells was significantly lower in the si-MeCP2-mimic group than that of the MeCP2-NC group and MeCP2-mimic group (both at P＜0.001).The relative expressing level of E-cadherin protein was significantly elevated and the expressions of β-catenin,Vimentin,MMP-9 and MMP-7 were significantly reduced in the si-MeCP2 group compared with the MeCP2-mimnic group and MeCP2-NC group (all at P＜O.01).The relative expressing level of SFRP5 protein in the MeCP2-mimic group,MeCP2-NC group and si-MeCP2 group was 27.19± 0.03,47.54±0.05 and 74.93±0.05,respectively,showing a statistical difference among the three groups (F =183.49,P＜0.001),and the relative expressing level of SFRP5 in the si-MeCP2 group was significantly higher than that in the MeCP2-mimic group and MeCP2-NC group (both at P＜0.001).Conclusions MeCP2C can promote EMT of human LECs by down-regulating the expression of SFRP5 and therefore activating the Wnt3a/β-catenin signal pathway.
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Objective:To explore the clinical effects of uncut Roux-en-Y anastomosis versus Roux-en-Y anastomosis in laparoscopic distal gastrectomy.Methods:The propensity score matching and retrospective cohort study was conducted. The clinicopathological data of 194 patients who underwent laparoscopic distal gastrectomy in the First Affiliated Hospital of Air Force Military Medical University from January 2017 to May 2019 were collected. There were 130 males and 64 females, aged (57±10)years, with a range from 27 to 78 years. Of 194 patients, 62 undergoing uncut Roux-en-Y anastomosis digestive tract reconstruction in laparoscopic distal gastrectomy and 132 undergoing Roux-en-Y anastomosis digestive tract reconstruction in laparoscopic distal gastrectomy were allocated into uncut group and traditional group, respectively. Observation indicators: (1) the propensity score matching conditions and comparison of general data between the two groups after propensity score matching; (2) intraoperative and postoperative situations; (3) follow-up. Follow-up using outpatient examination and telephone interview was conducted at the postoperative 3 months and 6 months to detect Roux stasis syndrome (RSS), tumor recurrence, readmission, and survival of patients. The propensity score matching was conducted by 1∶1 matching using the nearest neighbor method. Measurement data with normal distribution were represented as Mean± SD, and comparison between groups was done using the t test. Measurement data with skewed distribution were represented as M (range). Count data were represented as absolute numbers, and comparison between groups was analyzed using the chi-square test Fisher exact probability. Comparison of ordinal data between groups was analyzed using the nonparametric rank sum test. Results:(1) The propensity score matching conditions and comparison of general data between the two groups after propensity score matching: 104 of 194 patients had successful matching, including 52 in the uncut group and 52 in the traditional group respectively. Before propensity score matching, cases with age ≤60 years or >60 years, cases in stage Ⅰ, Ⅱ, Ⅲ of American Society of Anesthesiologists were 43, 19, 27, 28, 7 for the uncut group, respectively, versus 63, 69, 24, 92, 16 for the traditional group, showing significant differences between the two groups ( χ2=1.279, Z=2.818, P<0.05). After propensity score matching, the above indicators were 33, 19, 20, 25, 7 for the uncut group, versus 34, 18, 15, 33, 4 for the traditional group, showing no significant difference between the two groups ( χ2=0.000, Z=0.500, P>0.05). (2) Intraoperative and postoperative situations: 104 patients underwent laparoscopic distal gastrectomy successfully and received R 0 resection, without intraoperative complications or conversion to open surgery. After propensity score matching, the time of digestive tract reconstruction was (41±10)minutes for the uncut group, versus (52±15)minutes for the traditional group, showing a significant difference between the two groups ( t=4.511, P<0.05). (3) Follow-up: 104 patients were followed up at the postoperative 3 months and 6 months. The incidence of RSS at the postoperative 3 months was 0 for the uncut group, versus 30.8%(16/52) for the traditional group, showing a significant difference between the two groups ( P<0.05). The incidence of RSS at the postoperative 6 months was 0 for the uncut group, versus 9.6%(5/52) for the traditional group, showing no significant difference between the two groups ( P>0.05). There was no tumor recurrence, readmission, death within postoperative 30 days, or cancer-related death in the 104 patients. Conclusion:Uncut Roux-en-Y anastomosis is safe and feasible in the laparoscopic distal gastrectomy, which can effectively shorten the time of digestive tract reconstruction and reduce the occurrence of RSS after 3 months surgery.
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Objective:To investigate the role of IL-1β in regulating human placenta-derived mesenchymal stem cells (hPMSCs) to induce the differentiation of activated peripheral blood mononuclear cells (PBMCs) into CD8 + IL-10 + T cell subset in patients with rheumatoid arthritis (RA). Methods:Serum IL-1β levels in RA patients were detected by CBA kit. hPMSCs were isolated from healthy subjects by enzyme digestion. PBMCs that were isolated from RA patients and healthy subjects by Ficoll density gradient centrifugation were activated with PHA and CD3/CD28 monoclonal antibody (McAb) and then co-cultured with hPMSCs. Flow cytometry (FCM) was used to analyze the ability of hPMSCs to induce the differentiation of activated PBMCs into CD8 + IL-10 + T cell subset with the presence of programmed death ligand 1 (PD-L1) and/or PD-L2 McAb and IL-1β. Expression of PD-L1 and PD-L2 on hPMSCs under IL-1β stimulation was analyzed by RT-PCR and FCM. Results:Serum IL-1β levels were significantly higher in RA patients than in healthy subjects. After the activation by PHA and CD3/CD28 McAb, the proportion of CD8 + IL-10 + T cells in PBMCs of RA patients was significantly lower than that of healthy subjects. Moreover, the proportion of CD8 + IL-10 + T cells significantly increased after co-cultured with hPMSCs. The ability of IL-1β-treated hPMSCs to induce activated PBMCs to differentiate into CD8 + IL-10 + T cells was significantly enhanced compared to untreated hPMSCs. IL-1β could up-regulate the expression of PD-L1 and PD-L2 on hPMSCs. Blocking the expression of PD-L1 and/or PD-L2 on hPMSCs significantly reduced the ability of hPMSCs to induce activated PBMCs to differentiate into CD8 + IL-10 + T cells. Conclusions:IL-1β enhanced the capacity of hPMSCs to induce activated PBMCs to differentiate into CD8 + IL-10 + T cells in RA patients by up-regulating the expression of PD-L1 and PD-L2.
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@#Objective To compare the effect of different treatment strategies of vertical vein on the postoperative efficacy of total anomalous pulmonary venous connection (TAPVC) in newborns. Methods The clinical data of 72 TAPVC newborns who underwent correction in our hospital from June 2008 to January 2018 were retrospectively analyzed. There were 59 males and 13 females, with an average age of 9.93±8.13 d. According to different vertical vein treatment strategies, the patients were divided into an unligated group (22 patients), a partially ligated group (23 patients) and a completely ligated group (27 patients). The mortality and incidence of common complications among the groups were compared. Results In the completely ligated group, the aortic cross-clamp time was longer (P=0.001), intraoperative circulatory arrest cases were less (P=0.005), and the early velocity of pulmonary vein was faster (P=0.029), but there was no significant difference in the velocity of pulmonary vein for the last follow-up among three groups (P=0.393). There was no significant difference in other perioperative indicators among groups, including the early mortality, the incidence of pulmonary vein obstruction and the non-closure of vertical vein after surgery (P>0.05). The mean follow-up time was 4.47±2.63 years. The follow-up mortality was not significantly different among groups (P>0.05). The cardiac function (NYHA) of all the 64 survived patients was classⅠ-Ⅱ. Conclusion Proper vertical vein treatment strategies may be conducive to a smooth transition after surgery, ensuring the success of surgeries.
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Objective@#To investigate the role of methyl-CpG-binding protein 2 (MeCP2) in the regulation and epithelial-mesenchymal transition (EMT) of human lens epithelial cells (LECs) and its possible mechanism.@*Methods@#Human LEC lines (SRA01/04) were divided into MeCP2-mimic group, MeCP2-NC group and small interferening RNA-MeCP2 (si-MeCP2) group, and MeCP2 analog plasmid, blank plasmid and MeCP2 si-RNA plasmid was used respectively to transfect the cells.The expression of MeCP2 mRNA in the cells was detected by using real-time PCR 24 hours after transfection.At 48 hours after transfection, the migration rate of the cells was evaluated by scratching test, and the expression of Wnt3a protein in the cells was detected by immunofluorescence stainning.The relative expressions of β-catenin, E-cadherin, Vimentin, matrix metallo proteinase (MMP)-9, MMP-7 and secreted frizzled-related protein 5 (SFRP5) proteins in the cells were detected by Western blot.@*Results@#After 24 hours of transfection, the relative expression of MeCP2 mRNA in the cells was significantly different among the MeCP2-mimic group, MeCP2-NC group and si-MeCP2 group (F=4 773.00, P<0.00 1). The migrating rate of the cells in the MeCP2-mimic group, MeCP2-NC group and si-MeCP2 group was (57.45±5.20)%, (32.71±10.02)% and (17.77±9.22)%, respectively, showing a significant difference among the three groups (F=124.00, P<0.001), and the migrating rate of the cells in the si-MeCP2 group was significantly lower than that of the MeCP2-mimic group or MeCP2-NC group (both at P<0.001). The relative expressing intensity (absorbance) of Wnt3a in the cells of the MeCP2-mimic group, MeCP2-NC group and si-MeCP2 group was 75.92±6.10, 52.03±5.22 and 28.75±3.39, respectively, with a significant difference among three the groups (F=221.30, P<0.001), and the relative expressing intensity (absorbance) of Wnt3a in the cells was significantly lower in the si-MeCP2-mimic group than that of the MeCP2-NC group and MeCP2-mimic group (both at P<0.001). The relative expressing level of E-cadherin protein was significantly elevated and the expressions of β-catenin, Vimentin, MMP-9 and MMP-7 were significantly reduced in the si-MeCP2 group compared with the MeCP2-mimnic group and MeCP2-NC group (all at P<0.01). The relative expressing level of SFRP5 protein in the MeCP2-mimic group, MeCP2-NC group and si-MeCP2 group was 27.19±0.03, 47.54±0.05 and 74.93±0.05, respectively, showing a statistical difference among the three groups (F=183.49, P<0.001), and the relative expressing level of SFRP5 in the si-MeCP2 group was significantly higher than that in the MeCP2-mimic group and MeCP2-NC group (both at P<0.001).@*Conclusions@#MeCP2C can promote EMT of human LECs by down-regulating the expression of SFRP5 and therefore activating the Wnt3a/β-catenin signal pathway.