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Objective:To explore the methods of standardized residency training for internal medicine residents in the post-epidemic era, and to provide theoretical basis for improving and optimizing the standardized residency training.Methods:A total of 228 resident physicians from Batch 2017 to Batch 2019 were recruited, and a questionnaire survey was conducted to investigate their basic information, their attitudes and actions in facing the epidemic and the epidemic impact on the occupational planning and rotation plan. Original data of this study were exported through the questionnaire platform "Questionnaire Network", sorted out by Microsoft Excel, and plotted and analyzed by Origin software.Results:Residents of our hospital, socialized medical residents, other unit sponsor residents and combined professional masters willing to go to Hubei for medical support accounted for 100% (30 people), 86% (6 people), 84% (80 people) and 77% (72 people) respectively. Almost all of the residents were willing to learn the knowledge of COVID-19 and to educate the public (99%-100%). Eighty percent (24 people) of the residents of our hospital participated in anti-epidemic, while the proportion of other unit sponsor residents, socialized medical residents and combined professional masters were 46% (44 people), 14% (1 people) and 12% (11 people), respectively. Additionally, 97% (29 people) of our hospital residents, 89% (85 people) of other unit sponsor residents, 86% (6 people) of the socialized medical residents, and 82% (76 people) of the combined professional masters would still like to engage in the clinical work in the future. What's more, the resident physicians who thought that phylaxiology, epidemiology, general practice medicine, traditional Chinese medicine and critical care medicine should be strengthened to train accounted for 98% (221 people), 98% (221 people), 90% (203 people), 70% (158 people) and 60% (135 people) respectively.Conclusion:Most resident physicians have strong awareness and active actions in fighting against the epidemic. In the future, the standardized residency training of physicians should further strengthen the training of phylaxiology, epidemiology, general practice medicine, traditional Chinese medicine and critical care medicine.
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Objective:To explore the effects of SOAP (subjective objective assessment plan) case recording method by reflective teaching combined with the framework of International Classification of Functioning-Children and Youth Version (ICF-CY) in rehabilitation therapy practice.Methods:Interns of rehabilitation therapywere randomly divided into ICF-CY record group taking the case recording method of reflective teaching combined with the framework of ICF-CY and traditional record group taking traditional SOAP case recording method for children with cerebral palsy. After the internship, examination based on the clinical analysis and assessment of cases records and questionnaire survey were conducted among the two groups of interns. SPSS 20.0 was used to conduct t test and chi-square test. Results:The scores based on the clinical analysis and assessment of the ICF-CY record group were higher than those of the traditional record group. The questionnaire showed that interns' satisfaction of the SOAP case recording method by reflective teaching combined with the framework of ICF-CY was superior to the traditional SOAP case recording method ( P<0.001). Conclusion:The SOAP case recording method by reflective teaching combined with the framework of ICF-CY can improve interns' abilities of clinical case analysis, stimulate their self-learning abilities, and help to cultivate their thinking of clinical diagnosis and treatment.
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Objective:To investigate the effect of oxytocin on neonatal instinctive behavior expression and breast-feeding.Methods:A total of 71 pairs of parturient women and their newborns who delivered in the delivery room of the Affiliated Hospital of Weifang Medical College from February to May 2021 were selected. According to whether oxytocin induced labor, they were divided into the oxytocin group and the control group. Widstr?m nine stages was used to observe and record neonatal behavior. Breastfeeding was followed up 3 days and 3 months after delivery.Results:Totally 35 pairs in each group were included. In the oxytocin group, the occurrence time of neonatal head or head turning, hand in mouth, body moving to nipple, lips touching areola, licking nipple, nipple and effective sucking were (18.2 ± 5.4), (27.8 ± 8.3), (31.0 ± 10.1), (44.3 ± 14.5), (47.2 ± 15.6), (49.4 ± 16.3), (48.3 ± 13.6) min, which were significantly later than those in the control group (15.3 ± 5.3), (21.0 ± 8.1), (24.3 ± 9.0), (34.0 ± 11.4), (37.2 ± 11.9), (38.6 ± 11.8), (39.6 ± 8.7) min. The difference was statistically significant ( t values ranged from -3.10 to -2.17, P<0.05). The duration of neonatal awakening, activity, crawling and familiarity in the oxytocin group were (6.9 ± 3.2), (18.9 ± 9.3), (13.6 ± 7.9), (9.2 ± 5.1) min, which were significantly longer than those in the control group (5.1 ± 2.8), (12.3 ± 7.1), (10.3 ± 5.3), (6.7 ± 4.3) min; sucking stage duration in the oxytocin group was (35.1 ± 7.2) min, which was significantly shorter than that in the control group (39.6 ± 7.1) min; all the differences were statistically significant ( t values ranged from -3.25 to 2.28, P<0.05). The times of exclusive breast-feeding in the oxytocin group were (2.8 ± 3.1), (4.5 ± 3.3), (6.9 ± 3.0) at 24, 48 and 72 h postnatal, and the exclusive breastfeeding rate at 3 months after birth was 77.1%(27/35), which were significantly lower than those (7.6 ± 3.6), (8.9 ± 2.7), (10.3 ± 2.0) and 82.9%(29/35) in the control group, the differences were statistically significant ( t=6.05, 6.11, 5.48, χ2=0.36, P<0.05). Conclusions:The use of oxytocin during labor may affect the expression of neonatal lactation behavior and negatively affect the breastfeeding.
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To report a case of generalized lichen amyloidosis successfully treated with dupilumab in China. A 70-year-old male patient presented with extensive itchy papules on the trunk and extremities for 23 years. Skin examination revealed diffuse millet-to mung bean-sized hemispherical brown plaques on the trunk, right anterior shank, and extensor aspect of both upper arms, with a hard texture on palpation. No abnormalities were observed in the blood eosinophil count or serum IgE level. Histopathological examination of the skin lesion on the lower limb showed epidermal hyperkeratosis and homogeneous red-stained lumpy materials in the papillary dermis. Immunohistochemical study showed positive staining with Congo red. The pruritus numerical rating scale score was 10 points. The diagnosis of generalized lichen amyloidosis was confirmed. The patient received subcutaneous injection of dupilumab at an initial dose of 600 mg, followed by an every-2-week regimen at a dose of 300 mg. At week 2 after the start of treatment, pruritus was markedly relieved; at week 14, the skin lesions began to subside markedly; at week 18, the skin lesions on the chest and abdomen nearly completely subsided, and lesions on the lower back and limbs markedly regressed. No obvious adverse reactions were observed.
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[This corrects the article DOI: 10.1016/j.apsb.2021.09.017.].
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@#<b>Objective</b> To investigate the current situation of quality control in medical linear accelerators in Hebei Province, China. <b>Methods</b> The main performance indices of some medical linear accelerators in Hebei Province, 2019—2021 were tested according to the current relevant effective standards GB 15213—2016 <i>Medical Electron Accelerators-Functional Performance Characteristics and Test Methods</i> and WS 674—2020 <i>Specification for Testing of Quality Control in Medical Linear Accelerator</i>. <b>Results</b> A total of 175 medical linear accelerators were tested from 2019 to 2021, and the annual pass rates were 72.4% in 2019, 75.9% in 2020, and 79.4% in 2021. The overall pass rate of initial inspection was 76.0%, and the pass rate of reinspection was 100.0%. The index with the lowest pass rate in the initial inspection was square X radiation field uniformity, followed by dose deviation and square X radiation field symmetry. <b>Conclusion</b> Medical institutions should strengthen quality control management, conduct regular testing of medical linear accelerators, and timely correct dose deviation, so as to ensure the treatment effect of patients.
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Objective:To study the efficacy and influencing factors of ursodeoxycholic acid (UDCA) in the treatment of cholesterol gallstone, so as to provide reference for the treatment of cholesterol gallstone by internal medicine.Methods:From March 1, 2017 to March 31, 2018, at outpatient department of gastroenterology of 9 Beijing medical centers including Peking University People′s Hospital, the Sixth Medical Center of PLA General Hospital, Beijing Huaxin Hospital, PLA Rocket Force Characteristic Medical Center, Peking University Aerospace Center Hospital, Beijing Youan Hospital of Capital Medical University and Beijing Tiantan Hospital of Capital Medical University, Beijing Tongren Hospital of Capital Medical University, and Beijing Shijitan Hospital of Capital Medical University, the data of patients with cholesterol gallstone treated by UDCA were collected. The inclusion criteria were that the largest diameter of stone was ≤10 mm and the stone was not detected under X-ray. The treatment plan was taking UDCA orally for 6 months at a dose of 10 mg·kg -1·d -1. The basic information of patients, the ultrasound examination results before treatment and 6 months after treatment, and scores of biliary abdominal pain and dyspepsia symptom were collected. Univariate and multivariate logistic regression were used to analyze the influencing factors of the efficacy in gallstrone dissolution by UDCA, and Wilcoxon signed rank test was used for statistical analysis. Results:A total of 215 patients were enrolled. The complete dissolution rate of gallstone was 19.5% (42/215) and partial dissolution rate was 50.7% (109/215), and the total effective rate was 70.2% (151/215). The complete dissolution rate of sandy stone was significantly higher than that of lumped stones (37.0%(17/46) vs. 14.8%(25/169); OR=3.377, 95% confidence interval (95% CI) 1.621 to 7.035, P=0.001). In lumped stones, the complete dissolution rate of the stones with diameter ≤5 mm was significantly higher than that of the stones with diameter >5 mm (37.5%(9/24) vs. 11.0%(16/145); OR=4.837, 95% CI 1.823 to 12.839, P=0.002). The complete dissolution rate of patients with higher body mass index ( OR=0.872, 95% CI 0.764 to 0.995, P=0.043) and longer disease course ( OR=0.942, 95% CI 0.912 to 0.973, P<0.001) was low. The results of multivariate logistic analysis indicated that long disease course of gallstone ( OR=0.940, 95% CI 0.908 to 0.974, P=0.001), rough gallbladder wall ( OR=0.438, 95% CI 0.200 to 0.962, P=0.040) and lumped stone ( OR=0.236, 95% CI 0.101 to 0.550, P=0.001) were independent risk factors of influencing the efficacy of stone dissolution by UDCA. As for lumped stones, the independent risk factors included long disease course of gallstone ( OR=0.926, 95% CI 0.877 to 0.978, P=0.006) and stone diameter >5 mm ( OR=0.142, 95% CI 0.043 to 0.470, P=0.001). After 6 months of UDCA treatment, score of biliary abdominal pain decreased from 0 (0 to 6) to 0 (0 to 0) and the score of dyspepsia symptom decreased from 1 (0 to 2) to 0 (0 to 0), and the differences between before treatment and after treatment were statistically significant ( Z=-8.50, and -9.13, both P<0.001). Conclusions:UDCA has a certain efficacy in cholesterol gallstone dissolution and can ease biliary abdominal pain and dyspepsia symptom. Long disease course of gallstone, rough gallbladder wall and stone diameter >5 mm are independent risk factors of poor efficacy in gallstone dissolution by UDCA.
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Objective:To investigate the effect of KRT5 knockdown in keratinocytes on melanin content in co-cultured melanocytes, and to explain mechanisms underlying formation of hyperpigmented lesions in reticulate pigmented anomaly of the flexures (Dowling-Degos disease, DDD) .Methods:HaCaT cells with heterozygous mutations in the KRT5 gene were obtained by using clustered regularly interspaced short palindromic repeats (CRISPR) -CRISPR-associated protein 9 (Cas9) technology (experimental group) , and HaCaT cells transfected with non-targeting single guide RNA:Cas9 protein complex served as control group, both of which were in vitro co-cultured with primary human melanocyte cells (HEMn) separately. Immunofluorescence study was conducted to determine the expression of cytokeratin and melanosomes in co-cultured cells; melanin content was detected in melanocytes in different co-culture groups, which were obtained by differential trypsinization. Immunohistochemical study was performed to determine the expression of melanocyte-specific premelanosome protein 17 (Pmel17) in skin lesions in a patient with DDD carrying a KRT5 mutation and normal skin tissues in a healthy control. Results:Sanger sequencing showed a heterozygous mutation (c.1delA) at the initiation codon of exon 1 of the KRT5 gene in HaCaT cells in the experimental group, but no mutation in the KRT5 gene in the control group. Western blot analysis showed that the KRT5 protein expression was significantly lower in the experimental group (0.60 ± 0.05) than in the control group (1.00 ± 0.00, t = 32.38, P = 0.001) . Compared with the co-culture system in the control group, the number of Pmel17-labeled melanosomes markedly increased with the melanin content elevated by 52.5% ( t = -3.48, P = 0.025) in the HEMn cells co-cultured with HaCaT cells in the experimental group. Immunohistochemical study showed that the Pmel17 expression increased in the skin lesions in the DDD patient with KRT5 mutation compared with the normal skin tissues in the healthy control. Conclusion:The effect of HaCaT cells with CRISPR-Cas9-induced KRT5 mutation on the co-cultured HEMn melanocytes was verified by the successfully established in vitro co-culture system, which provides a primary cell model for further studies on interaction mechanisms between keratinocytes and melanocytes, and on pathogenesis of skin pigmentation abnormalities.
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Objective:To investigate the application value of Metagenomic Next-Generation sequencing (mNGS) in infectious patients after allogeneic hematopoietic stem cell transplantation(allo-HSCT).Methods:Patients suspected with local or systemic infections were retrospectively included after allo-HSCT in our department from April 2019 to November 2020. Pathogenic microorganisms were tested by mNGS in samples from peripheral blood, cerebrospinal fluid, alveolar lavage Liquid, abscess, etc. Other diagnostic methods such as bacterial/fungal culture, viral PCR detection were simultaneously explored comparing with mNGS results.Results:A total of 112 samples in 83 patients were detected by mNGS, and 34 pathogenic microorganisms were determined. Among these positive samples, 11 strains of bacteria (17 times) with the most common Escherichia coli (4/17) were reported. There were 7 strains of fungi (10 times) detected with primary Candida albicans (7/29). Although arvovirus 30.2% (39/129) were predominantly detected, its diagnostic relevance with infections was not definite. Other pathogenic viruses including cytomegalovirus (CMV) 25.6% (33/129) and Epstein Barr virus (EBV) 14.0% (18/129)were of significance. Comparing with golden diagnostic criteria, the sensitivity of mNGS was 86.5%, and specificity was 45.0%. Regarding single pathogen infection, the consistency of mNGS and conventional methods was 82.9% (29/35), while it was 16/17 in combination infections.Conclusion:mNGS could be a potential method to determine pathogens in patients suspected with infections after allo-HSCT.
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Radiation-induced brain injury (RBI) is one of the complications after radiotherapy for head and neck malignant tumors, which seriously affects the quality of life of patients. The pathophysiological mechanism of RBI is not completely clear. Current studies suggest that it is involved in a variety of cells in the central nervous system (CNS), whereas astrocyte, as the largest number of glial cells in the CNS, plays an important role in maintaining the CNS homeostasis and responding to CNS injury. In this article, the role of astrocytes in RBI was reviewed.
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Heterogeneous nuclear rib nucleoproteins belong to a class of RNA-binding proteins and possess highly conserved structures. Among them, the heterogeneous nuclear rib nucleoprotein A2/B1 (heterogeneous nuclear rib nucleoprotein A2, hnRNPA2/B1) is involved in many processes such as transcription and translation. In recent years, in the field of neurodegenerative diseases, it has been found that hnRNPA2/B1 promotes the accumulation of fibrin in the cytoplasm and participates in the formation of stress granules in neurons, which involves a variety of processes related to cognitive function, and is closely related to neurodegenerative diseases such as amyotrophic lateral sclerosis, frontotemporal dementia, Alzheimer′s disease and so on. This article reviews the related research, summarizes the important role and mechanism of hnRNPA2/B1 in neurodegenerative diseases, and provides help for the diagnosis and treatment of neurodegenerative diseases in the future.
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Objective:To find new biomarkers for the diagnosis and prognosis of sepsis through analyzing the differential expression protein in sepsis by proteomics and bioinformatics analysis and enzyme linked immunosorbent assay (ELISA).Methods:Patients with sepsis admitted to the emergency department of the Affiliated Hospital of Southwest Medical University from January to December 2019 were enrolled. And meanwhile, healthy volunteers who had normal physical examinations were included as the control group. Blood samples from two groups were collected. The samples were randomly selected for the protein concentration by data independent acquisition (DIA). Bioinformatics method was used in differentially expressed proteins by gene ontology (GO) pathway, enrichment analyses, groups meta-analysis and survival curves construction. ELISA method was used to verified marker screened. Then the data of transferrin receptor CD71 and the clinical data of procalcitonin (PCT), C-reactive protein (CRP) and blood lactic acid (Lac) were collected to construct receiver operator characteristic curve (ROC curve), and biomarker was screened for diagnostic and prognostic of sepsis.Results:The result of DIA showed that 71 differentially expressed proteins were screened out from sepsis group, 6 proteins were down-regulated and 65 proteins were up-regulated. Those differentially expressed proteins were enriched in the inflammatory response, response to stress, leukocyte migration in the GO pathway and enrichment analyses. The meta-analysis showed that the expression level of CD71 was higher in sepsis group than normal control group [standardized mean difference ( SMD) = -0.47, 95% confidence interval (95% CI) was -0.93 to 0.00, P < 0.01], the expression level of CD71 was higher in non-survivor group than survivor group ( SMD = -0.44, 95% CI was -0.70 to -0.18, P = 0.63). Survival curve showed that the expression of CD71 was inversely correlated to survival rates, the patients with a lower expression had higher survival rates ( P = 0.000 34); the ELISA showed that the level of CD71 was higher in sepsis group than normal control group (nmol/L: 156.83±84.71 vs. 87.99±47.89, P < 0.05), the level of CD71 was higher in non-survivor group than survivor group (nmol/L: 219.63±125.59 vs. 130.97±40.45, P < 0.05). The area under the ROC curve (AUC) of CD71 in diagnostic performance of sepsis was 0.790 (sensitivity was 65.1%, specificity was 90.0%), the AUC of CD71 in prognostic performance of sepsis was 0.744 (sensitivity was 57.1%, specificity was 94.1%); CD71 had a better prognostic performance than PCT (AUC = 0.547, sensitivity was 64.3%, specificity was 55.9%), CRP (AUC = 0.594, sensitivity was 64.3%, specificity was 61.8%), Lac (AUC = 0.540, sensitivity was 42.9%, specificity was 82.4%). Conclusion:CD71 had a great value of diagnostic and prognostic performance in sepsis, and it was expected to be a potential biomarker for sepsis.
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Objective:To translate the Pediatric Respiratory Assessment Measure into Chinese, and to test its reliability and validity.Methods:According to the Brislin translation model, the English version of Pediatric Respiratory Assessment Measure was translated into Chinese, and then was back-translated and modified for cultural adaptation. From June 2019 to December 2020, convenience sampling method was used to evaluate the severity of 50 children with acute attack of bronchial asthma aged 2-14 years in the Affiliated Huaian No.1 People′s Hospital of Nanjing Medical University, in order to evaluate its reliability and validity.Results:The Cronbach α of the Chinese version of Pediatric Respiratory Assessment Measure was 0.838, the inter-rater reliability was 0.984; the average scale-content validity index was 0.97. One common factor was extracted, the cumulative variance contribution rate of superior sternal fossa depression was 61.83%. The criterion related validity between the evaluation results of doctors, nurses, respiratory therapists and the grading standard of bronchial asthma severity in the " Guidelines for the diagnosis, prevention and treatment of bronchial asthma in children ( 2016 edition)" were 0.903, 0.884 and 0.889, the ROC curve of the scale was 0.867 (95% CI 0.754-0.980). Conclusions:The Chinese version of Pediatric Respiratory Assessment Measure has good reliability and validity, which can be used to evaluate the severity of acute attack of bronchial asthma in children aged 2-14 in China, and provide a basis for further diagnosis and treatment.
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Objective:To investigate the effects of early Comfort using Analgesics, minimal Sedatives and maximum Humane care (eCASH) patterns on the risk of negative mood and continuous renal replacement therapy (CRRT)-related adverse events in patients with severe CRRT.Methods:A total of 90 patients with severe CRRT in Shenzhen Integrated Traditional Chinese and Western Medicine Hospital from October 2018 to October 2020 were selected as the study subjects, and they were divided into observation group and control group according to random number table method, with 45 patients in each group. The control group was given routine nursing program, and the observation group was given eCASH mode on the basis of the control group.Nursing satisfaction, CRRT-related adverse events and negative mood scores before and after nursing were compared between 2 groups.Results:The nursing satisfaction degree of the observation group was 86.67% (39/45), significantly higher than that of the control group (66.67%, 30/45), and the difference was statistically significant ( χ2=5.03, P<0.05). After nursing, the Hospital Anxiety and Depression Scale-Anxiety(HADS-A) and Hospital Anxiety and Depression Scale-Depression(HADS-D) scores of the observation group were 5.18 ± 0.67 and 5.27 ± 0.61, respectively, lower than 8.14 ± 1.18, 7.94 ± 1.07 before intervention, and 6.33 ± 0.72, 5.94 ± 0.49 of the control group. Barthel Index(BI) (65.17 ± 8.67) was significantly higher than that before nursing 41.56 ± 6.46 and control group 60.48 ± 6.47, the difference was statistically significant ( t values were 5.74-20.76, all P<0.05). The scores of Visual Analogue Scale(VAS), Present Pain Intensity(PPI), sensory total score and emotional total score of observation group after nursing were 3.24 ± 0.56, 1.18 ± 0.25, 6.38 ± 0.89, 2.68 ± 0.59 significantly lower than those before nursing 6.24 ± 0.87, 3.24 ± 0.56, 11.24 ± 1.81, 6.37 ± 1.04 and 4.36 ± 0.67, 1.31 ± 0.31, 7.26 ± 0.96, 2.98 ± 0.62 of the control group. The difference was statistically significant ( t values were 2.19-20.70, P<0.05). Conclusions:eCASH model can significantly improve the negative emotions of patients with severe CRRT, improve their comfort and reduce the risk of related adverse events, which is worthy of clinical promotion.
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Pulmonary endothelial barrier dysfunction is a hallmark of clinical pulmonary edema and contributes to the development of acute lung injury (ALI). Here we reported that ruscogenin (RUS), an effective steroidal sapogenin of Radix Ophiopogon japonicus, attenuated lipopolysaccharides (LPS)-induced pulmonary endothelial barrier disruption through mediating non-muscle myosin heavy chain IIA (NMMHC IIA)‒Toll-like receptor 4 (TLR4) interactions. By in vivo and in vitro experiments, we observed that RUS administration significantly ameliorated LPS-triggered pulmonary endothelial barrier dysfunction and ALI. Moreover, we identified that RUS directly targeted NMMHC IIA on its N-terminal and head domain by serial affinity chromatography, molecular docking, biolayer interferometry, and microscale thermophoresis analyses. Downregulation of endothelial NMMHC IIA expression in vivo and in vitro abolished the protective effect of RUS. It was also observed that NMMHC IIA was dissociated from TLR4 and then activating TLR4 downstream Src/vascular endothelial cadherin (VE-cadherin) signaling in pulmonary vascular endothelial cells after LPS treatment, which could be restored by RUS. Collectively, these findings provide pharmacological evidence showing that RUS attenuates LPS-induced pulmonary endothelial barrier dysfunction by inhibiting TLR4/Src/VE-cadherin pathway through targeting NMMHC IIA and mediating NMMHC IIA‒TLR4 interactions.
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Contributing to organ formation and tissue regeneration, extracellular matrix (ECM) constituents provide tissue with three-dimensional (3D) structural integrity and cellular-function regulation. Containing the crucial traits of the cellular microenvironment, ECM substitutes mediate cell-matrix interactions to prompt stem-cell proliferation and differentiation for 3D organoid construction in vitro or tissue regeneration in vivo. However, these ECMs are often applied generically and have yet to be extensively developed for specific cell types in 3D cultures. Cultured cells also produce rich ECM, particularly stromal cells. Cellular ECM improves 3D culture development in vitro and tissue remodeling during wound healing after implantation into the host as well. Gaining better insight into ECM derived from either tissue or cells that regulate 3D tissue reconstruction or organ regeneration helps us to select, produce, and implant the most suitable ECM and thus promote 3D organoid culture and tissue remodeling for in vivo regeneration. Overall, the decellularization methodologies and tissue/cell-derived ECM as scaffolds or cellular-growth supplements used in cell propagation and differentiation for 3D tissue culture in vitro are discussed. Moreover, current preclinical applications by which ECM components modulate the wound-healing process are reviewed.
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Cell Differentiation , Cell Proliferation , Decellularized Extracellular Matrix , Extracellular Matrix/metabolism , Humans , Mesenchymal Stem Cells , Tissue Engineering/methods , Tissue Scaffolds/chemistryABSTRACT
ObjectiveThe effect of modified Shengjiangsan on immunoglobulin A （IgA） nephropathy was observed. The microRNA-148b （miRNA-148b）， interleukin 6 （IL-6）， core 1 beta 1，3-galactosyltransferase （C1GALT1）， molecular chaperone Cosmc （core1β3-Gal-T-specific molecular chaperone C1GALT1C1）， and galactose-deficient IgA1 （Gd-IGA1） in serum and kidney tissues of IgA nephropathy rats were detected to explore the underlying mechanism. The result is expected to lay a scientific basis for clinical application of modified Shengjiangsan in the treatment of IgA nephropathy. MethodA total of 42 SPF male SD rats were randomized into the normal group （8rats） and modeling group （34 rats） with the random number table method. After one week of adaptive feeding， rats for modeling were given bovine serum albumin （BSA， gavage）， lipopolysaccharide （LPS， injection into tail vein）， carbon tetrachloride （CCl4， subcutaneous injection）， and castor oil to induce IgA nephropathy. After modeling， two rats were randomly selected to test the modeling outcome. Then the model rats were classified into the model group， low-dose Chinese medicine group （modified Shengjiangsan，6.27 g·kg-1）， high-dose Chinese medicine group （modified Shengjiangsan，12.54 g·kg-1）， and benazepril group （10 mg·kg-1） with the random number table method， 8 in each group. The administration （gavage， once a day） lasted 4 weeks. The 24-h urinary total protein （24 h-UTP） was detected at the end of the 1st， 9th， and 13th week of the experiment. At the 14th week， after anesthesia， femoral artery blood was collected and centrifugated. The supernatant was collected to detect albumin （ALB）， aspartate aminotransferase （AST）， alanine aminotransferase （ALT）， serum creatinine （SCr）， and blood urea nitrogen （BUN）. The expression levels of IL-6 and Gd-IGA1 were determined by enzyme-linked immunosorbent assay （ELISA）. Based on hematoxylin-eosin （HE）/Masson/periodic Schiff-methenamine silver （PASM） staining， the pathological changes of renal tissues were observed. Ultrastructural changes of glomeruli were observed by transmission electron microscopy. The expression of miRNA-148b， IL-6， C1GALT1， and C1GALT1C1 was detected by immunohistochemistry. The mesangial area of the glomeruli was observed by immunofluorescence. Real-time polymerase chain reaction （Real-time PCR） was employed to determine the mRNA levels of mirNA-148b， IL-6， C1GALT1， and C1GALT1C1， and Western blot was used to detect the protein levels of IL-6， C1GALT1， and C1GALT1C1. ResultCompared with normal group， the model group showed increase in the content of 24 h-UTP， SCr， ALT， IL-6， and GD-IGA1 （P<0.05）， decrease in ALB content （P<0.05）. Moreover， rats in the model group demonstrated hyperplasia of glomerular mesangial cells， thickening of mesangial area， podocyte foot process effacement， and a large number of granular IgA immune complex in the mesangial area. In addition， the model group showed increase in the expression of IL-6 in mesangial area and podocytes， decrease in the expression of C1GALT1 and C1GALT1C1 in mesangial area and podocytes， enhanced expression of IL-6 mRNA and miRNA-148b （P<0.01）， weakened expression of C1GALT1 mRNA and C1GALT1C1 mRNA （P<0.01）， rise of IL-6 protein expression （P<0.01）， and reduction in the protein expression of C1GALT1 and C1GALT1C1 （P<0.01）. Compared with the model group， modified Shengjiangsan decreased the content of 24 h-UTP， SCr， ALT， IL-6， and Gd-IGA1 （P<0.05） and increased the content of ALB （P<0.05， P<0.01）. Moreover， with the treatment of this Chinese medicine， the pathological damage was significantly alleviated and the deposition of IgA immune complex in basement membrane was reduced. The expression of IL-6 in the mesangial area and podocytes of rats was decreased， and the expression of C1GALT1 and C1GALT1C1 in the mesangial area and podocytes of rats was increased. Moreover， the expression of IL-6 mRNA and miRNA-148b was decreased （P<0.01）， and the expression of C1GALT1 mRNA and C1GALT1C1 mRNA was increased （P<0.01）. The protein expression of IL-6 was decreased （P<0.05， P<0.01）， and the protein expression of C1GALT1 and C1GALT1C1 was enhanced （P<0.05， P<0.01）. The Chinese medicine group showed obvious dose-effect trend. ConclusionModified Shengjiangsan may reduce the expression of miRNA-148b and IL-6 in serum and kidney tissue of IgA nephropathy rats， restore the expression of C1GALT1 and C1GALT1C1， and decrease the generation of Gd-IGA1， so as to reduce renal pathological damage and proteinuria， protect the kidney protection， and finally delay the disease progression. Moreover， the effect is enhanced with the rise of dose.
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Objective:To establish a presenilin enhancer-2 (PSENEN) gene-silenced human immortalized keratinocyte (HaCaT) cell model, and to evaluate the effect of PSENEN gene silencing on the proliferation of and γ-secretase expression in HaCaT cells.Methods:Three shRNAs targeting the PSENEN gene were constructed, and inserted into the linearized LV3-pGLV-h1-GFP-puro vector to establish a recombinant lentiviral expression plasmid. After restriction enzyme digestion and sequencing, lentiviral packaging and purification were performed, and lentiviral titer was determined. Cultured HaCaT cells were divided into 5 groups: shRNA1, shRNA2 and shRNA3 groups treated with the lentivirus solutions containing PSENEN gene-targeted shRNA1, shRNA2 and shRNA3 respectively, NC group treated with the lentivirus solution containing a negative control shRNA (shNC) , and blank group treated without lentivirus solution. After transfection, inverted fluorescence microscopy was performed, and transfection efficiency was determined by flow cytometry. Cell counting kit-8 (CCK8) assay was performed to evaluate the effect of PSENEN gene silencing on the proliferation of HaCaT cells, and real-time fluorescence-based quantitative PCR (qPCR) and Western blot analysis were conducted to determine the mRNA and protein expression of PSENEN, nicastrin (NCT) , presenilin-1 (PS1) and anterior pharynx defective 1a (APH1a) genes respectively. Statistical analysis was carried out by using repeated measures analysis of variance, one-way analysis of variance, and least significant difference t test for multiple comparisons. Results:Inverted fluorescence microscopy showed that fluorescence was observed in the shRNA1 group, shRNA2 group, shRNA3 group and NC group, and flow cytometry showed that the transfection efficiency was over 98% in the above 4 groups. qPCR and Western blot analysis revealed that the mRNA and protein expression of PSENEN gene significantly decreased in the shRNA1 (0.187 ± 0.010, 0.219 ± 0.097, respectively) , shRNA2 (0.163 ± 0.022, 0.208 ± 0.014, respectively) and shRNA3 (0.174 ± 0.009, 0.185 ± 0.062, respectively) groups compared with the NC group (1.054 ± 0.272, 1.076 ± 0.075, respectively, all P < 0.001) . CCK8 assay showed that the cellular proliferative activity significantly increased in the shRNA1 group compared with the NC group at 0, 12, 36 and 48 hours (all P < 0.05) , and there was no significant difference between the 2 groups at 24 or 60 hours (both P > 0.05) ; the cellular proliferative activity was significantly higher in the shRNA2 and shRNA3 groups than in the NC group at 0, 12, 24, 36, 48 and 60 hours (all P < 0.05) . There was no significant difference in the mRNA expression of NCT, PS1 and APH1a genes among the shRNA1 group, shRNA2 group, shRNA3 group, NC group, and blank group ( F= 8.168, 4.644, 1.981, respectively, all P > 0.05) , while the relative protein expression level of mature NCT (mNCT) , immature NCT (imNCT) , carboxyl-terminal fragment of PS1 (PS1-CTF) and APH1a significantly differed among the above 5 groups ( F= 39.268, 5.929, 27.842, 20.663, respectively, all P ≤ 0.01) . Compared with the NC group, the shRNA1, shRNA2 and shRNA3 groups all showed significantly decreased protein expression of mNCT, PS1-CTF and APH1a (all P < 0.01) , but insignificant changes in imNCT protein expression (all P > 0.05) . Conclusion:The PSENEN gene-silenced HaCaT cell model was successfully constructed, and the PSENEN gene silencing could lead to an increase in the cellular proliferative activity of HaCaT cells and a decrease in the protein expression of γ-secretase subunits mNCT, PS1-CTF and APH1a.
ABSTRACT
Objective:In general, patients with seropositive rheumatoid arthritis (RA) are considered to show an aggressive disease course. However, the relationship between the two subgroups in disease severity is controversial. Our study is aimed to compare the clinical characteristics and prognosis of double-seropositive and seronegative RA in China through a real-world large scale study.Methods:RA patients who met the 1987 American College of Rheumatology (ACR) classification criteria or the 2010 ACR/European Anti-Rheumatism Alliance RA classification criteria, and who attended the 10 hospitals across the country from September 2015 to January 2020, were enrolled. According to the serological status, patients were divided into 4 subgroups [rheumatoid factor (RF)(-) anti-cyclic citrullinated peptide (CCP) antibody (-), RF(+), RF(+) anti-CCP antibody(+), anti-CCP antibody(+)] and compared the disease characteristics and treatment response. One-way analysis of variance was used for measurement data that conformed to normal distribution, Kruskal-Wallis H test was used for measurement data that did not conform to normal distribution; paired t test was used for comparison before and after treatment within the group if the data was normally distributed else paired rank sum test was used; χ2 test was used for count data. Results:① A total of 2 461 patients were included, including 1 813 RF(+) anti-CCP antibody(+) patients (73.67%), 129 RF(+) patients (5.24%), 245 RF(-) anti-CCP antibody(-) patients (9.96%), 74 anti-CCP antibody(+) patients (11.13%). ② Regardless of the CCP status, RF(+) patients had an early age of onset [RF(-) anti-CCP antibody(-) (51±14) years old, anti-CCP antibody(+) (50±15) years old, RF(+) anti-CCP antibody(+) (48±14) years old, RF(+)(48±13) years old, F=3.003, P=0.029], longer disease duration [RF(-) anti-CCP antibody(-) 50 (20, 126) months, anti-CCP antibody(+) 60(24, 150) months, RF(+) anti-CCP antibody(+) 89(35, 179) months, RF(+) 83(25, 160) months, H=22.001, P<0.01], more joint swelling counts (SJC) [RF(-) anti-CCP antibody(-) 2(0, 6), Anti-CCP antibody(+) 2(0, 5), RF(+) anti-CCP antibody(+) 2(0, 7), RF(+) 2(0, 6), H=8.939, P=0.03] and tender joint counts (TJC) [RF(-) anti-CCP antibody(-) 3(0, 8), anti-CCP antibody(+) 2(0, 6), RF(+) anti-CCP antibody(+) 3(1, 9), RF(+) 2(0, 8), H=11.341, P=0.01] and the morning stiff time was longer [RF(-) anti-CCP antibody(-) 30(0, 60) min, anti-CCP antibody(+) 20(0, 60) min, RF(+) anti-CCP antibody(+) 30(10, 60) min, RF(+) 30(10, 60) min, H=13.32, P<0.01]; ESR [RF(-) anti-CCP antibody(-) 17(9, 38) mm/1 h, anti-CCP antibody(+) 20(10, 35) mm/1 h, RF(+) anti-CCP antibody(+) 26(14, 45) mm/1 h, RF(+) 28(14, 50) mm/1 h, H=37.084, P<0.01] and CRP [RF(-) anti-CCP antibody(-) 2.3 (0.8, 15.9) mm/L, Anti-CCP antibody(+) 2.7(0.7, 12.1) mm/L, RF(+) anti-CCP antibody(+) 5.2(1.3, 17.2) mm/L, RF (+) 5.2(0.9, 16.2) mm/L, H=22.141, P<0.01] of the RF(+)patients were significantly higher than RF(-) patients, and RF(+) patients had higher disease severity(DAS28-ESR) [RF(-) anti-CCP antibody(-) (4.0±1.8), anti-CCP antibody(+) (3.8±1.6), RF(+) anti-CCP antibody(+) (4.3±1.8), RF(+) (4.1±1.7), F=7.269, P<0.01]. ③ The RF(+) anti-CCP antibody(+) patients were divided into 4 subgroups, and it was found that RF-H anti-CCP antibody-L patients had higher disease severity [RF-H anti-CCP antibody-H 4.3(2.9, 5.6), RF-L anti-CCP antibody-L 4.5(3.0, 5.7), RF-H anti-CCP antibody-L 4.9(3.1, 6.2), RF-L anti-CCP antibody-H 2.8(1.8, 3.9), H=20.374, P<0.01]. ④ After 3-month follow up, the clinical characteristics of the four groups were improved, but there was no significant difference in the improvement of the four groups, indicating that the RF and anti-CCP antibody status did not affect the remission within 3 months. Conclusion:Among RA patients, the disease activity of RA patients is closely related to RF and the RF(+) patients have more severe disease than RF(-) patients. Patients with higher RF titer also have more severe disease than that of patients with low RF titer. After 3 months of medication treatment, the antibody status does not affect the disease remission rate.
ABSTRACT
Objective:To explore the intervention effects of high and low audio-motor stimulation mode on attention and memory of elderly with mild cognitive impairment in rural areas.Methods:From September 2019 to May 2020, 80 subjects with mild cognitive impairment meeting the inclusion and exclusion criteria were selected and randomly divided into experimental group and control group according to the drawing lots, with 40 participants in each group.The experimental group was given high and low audio-motor stimulation mode training, and the control group was given finger exercises training.Before and after the intervention, sustained and selective attention were assessed using number cancellation test(NCT), E-Prime paradigm based on Stroop effect design, and memory was assessed using the Rivermead behavioral memory test, version 2 (RBMT-Ⅱ). SPSS 17.0 software was used to perform data statistics. The independent sample t test, paired sample t test and Mann Whitney U test were used for statistical analysis. Results:After intervention, the sustained attention index of the experimental group (0.86±0.48) was significantly higher than that of the control group (0.63±0.47, t=4.336, P<0.05). After intervention, the unanimous response time of the experimental group ((2 263.38±437.87) ms) and the reverse reaction time ((2 335.75±599.86) ms) were significantly lower than those of the control group ((2 703.95±506.34) ms, (2 667.43±585.38) ms) ( t=-4.068, -2.447, P<0.05). The consistent correct rate ((0.84±0.14)) and the reverse correct rate ((0.87±0.10))were significantly higher than those of the control group ((0.71±0.19), (0.73±0.15)) ( t= 3.571, 4.931, both P<0.05). RBMT-Ⅱ dimensions and total standard scores of the experimental group (19.00 (18.00, 20.00)) were higher than those of the control group (11.00 (10.00, 12.00))( Z=7.299, P<0.05). Conclusion:High and low audio-motor stimulation improves attention and memory in rural elderly with mild cognitive impairment.