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Objective:To investigate effects of mesalazine(MS)on proliferation,apoptosis and inflammatory injury of cell model of ulcerative colitis(UC)induced by lipopolysaccharide(LPS),as well as transforming growth factor-β1(TGF-β1)/Smad signaling pathway effect in this study.Methods:Human colonic epithelial cells NCM-460 cultured in vitro were induced UC model by LPS,and divided into Con group(no treatment),LPS group(1 mg/L LPS),MS group(0.1,0.2,0.4 mg/L MS+1 mg/L LPS)and inhibitor group(10 μmol/L TGF-β1/Smad signaling pathway inhibitor LY2109761+0.2 mg/L MS+1 mg/L LPS).Cell morphology,proliferation,apoptosis and levels of inflammatory factors and TGF-β1/Smad pathway-related markers were examined by inverted microscope,EdU assay,Hoechst 33258 staining,ELISA and Western blot.Results:LPS treatment highly induced cell proliferation rate and Smad7 pro-tein level compared with Con group,while apoptotic cells,inflammatory factors TNF-α and IL-6,soluble interleukin-2 receptor(sIL-2R)release,as well as TGF-β1,p-Smad2,p-Smad3 protein expressions were increased;the above effects induced by LPS was reversed by MS in a dose-dependent manner(P<0.05).Compared with 0.2 mg/L MS group,NCM-460 cells proliferation rate and Smad7 expression were increased,while apoptotic cells,TNF-α and IL-6,sIL-2R releases,and TGF-β1,p-Smad2,p-Smad3 protein expressions were decreased(P<0.05).Conclusion:MS can attenuate LPS-induced apoptosis and inflammatory injury in NCM-460 cells,and this protection was possibly through suppressing TGF-β1/Smad signaling pathway.
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Skin regeneration is hindered by poor vascularization, prolonged inflammation, and excessive scar tissue formation, which necessitate newer strategies to simultaneously induce blood vessel regeneration, resolve inflammation, and induce host cell recruitment. Concurrent deployment of multiple biological cues to realize synergistic reparative effects may be an enticing avenue for wound healing. Herein, we simultaneously deployed SDF (stromal cell-derived factor)- 1α, VEGF (vascular endothelial growth factor)-binding peptide (BP), and GLP (glucagon like peptide)- 1 analog, liraglutide (LG) in core/shell poly(L-lactide-co-glycolide)/gelatin fibers to harness their synergistic effects for skin repair in healthy as well as diabetic wound models in rats. Microscopic techniques, such as SEM and TEM revealed fibrous and core/shell type morphology of membranes. Boyden chamber assay and scratch-wound assay displayed significant migration of HUVECs (human umbilical vein endothelial cells) in SDF-1α containing fibers. Subcutaneous implantation of membranes revealed higher cellular infiltration in SDF-1α loaded fibers, especially, those which were co-loaded with LG or BP. Implantation of membranes in an excisional wound model in healthy rats further showed significant and rapid wound closure in dual cues loaded groups as compared to control or single cue loaded groups. Similarly, the implantation of dressings in type 2 diabetes rat model revealed fast healing, skin appendages regeneration, and blood vessel regeneration in dual cues loaded fibers (SDF-1α/LG, SDF-1α/BP). Taken together, core/shell type fibers containing bioactive peptides significantly promoted wound repair in healthy as well as diabetic wound models in rats.
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Diabetes Mellitus Experimental , Diabetes Mellitus Tipo 2 , Ratas , Humanos , Animales , Gelatina/farmacología , Señales (Psicología) , Factor A de Crecimiento Endotelial Vascular/metabolismo , Quimiocina CXCL12/farmacología , Cicatrización de Heridas , Células Endoteliales de la Vena Umbilical Humana/metabolismo , InflamaciónRESUMEN
ObjectiveTo investigate the effect of Glycyrrhizae Radix et Rhizoma (GR)-containing serum on lipopolysaccharide (LPS)-induced inflammation in human colon epithelial adenocarcinoma cells (Caco2) based on inhibition of ferroptosis by the nuclear factor erythroid 2-related factor 2 (Nrf2)/heme oxygenase-1 (HO-1) pathway. MethodCaco2 cells were divided into a normal group, a model group (LPS, 200 μg·L-1), low-, medium-, and high-dose GR-containing serum groups (5%, 10%, 20%), and a ferroptosis inhibitor group (3-amino-4-cyclohexylamino-benzoic acid ethyl ester, Fer-1, 10 μmol·L-1). The cells in the normal group were cultured normally, while those in other groups underwent the induction of an inflammation model. The cells in the low-, medium-, and high-dose GR-containing serum groups were treated with 5%, 10%, and 20% GR-containing serum for 24 hours, respectively, and the cells in the ferroptosis inhibitor group were treated with Fer-1 for 24 hours. Transmission electron microscopy was used to observe mitochondrial morphology in each group. Flow cytometry was used to detect intracellular Fe2+ levels. Microplate assays were performed to measure superoxide dismutase (SOD) activity, malondialdehyde (MDA) and glutathione peroxidase (GSH-Px) levels. Enzyme-linked immunosorbent assay (ELISA) was used to measure interleukin-1β (IL-1β), IL-6, IL-10, and tumor necrosis factor-α (TNF-α) levels. Western blot was used to measure the expression levels of Nrf2, HO-1, ferritin heavy chain 1 (FTH1), and glutathione peroxidase 4 (GSH-Px4) proteins. Small interfering RNA (siRNA) was used to investigate the role of Nrf2 in ferroptosis regulation. The cells after interference were divided into a negative control (NC) group, a Si-Nrf2 group, a GR-containing serum (20%) + Si-Nrf2 group, and a GR-containing serum (20%) + NC group. Microplate assays were performed to measure MDA, SOD, and GSH-Px levels, and Western blot was used to measure the expression levels of Nrf2, HO-1, FTH1, and GSH-Px4 proteins. ResultCompared with the normal group, the model group showed mitochondrial contraction, increased mitochondrial membrane thickness, and smaller mitochondrial morphology, increased Fe2+ content (P<0.01), blunted SOD activity (P<0.01), decreased GSH-Px expression (P<0.01), increased MDA content (P<0.01), reduced expression levels of Nrf2 and HO-1 (P<0.05), reduced FTH1 expression (P<0.01), and down-regulated GSH-Px4 expression (P<0.01). In the GR-containing serum groups, the medium- and high-dose groups showed a significant decrease in Fe2+ content (P<0.01), potentiated SOD and GSH-Px activities (P<0.01), and decreased MDA levels (P<0.01). The high-dose group showed a significant increase in Nrf2 expression (P<0.05), and the medium-dose group showed increased expression of HO-1 and GSH-Px4 proteins (P<0.05). The expression levels of FTH1 significantly increased in the low-, medium-, and high-dose groups (P<0.01). The study on mechanism revealed that compared with the NC group, the cells transfected with Nrf2 siRNA showed increased MDA content (P<0.01), blunted SOD activity (P<0.01), decreased GSH-Px activity (P<0.01), decreased expression of Nrf2 and HO-1 (P<0.01), and reduced levels of FTH1 and GSH-Px4 proteins (P<0.01). Compared with the Si-Nrf2 group, the cells treated with GR-containing serum showed a decrease in MDA content (P<0.01), an increase in SOD activity (P<0.01), an increase in GSH-Px activity (P<0.01), increased expression of Nrf2 and FTH1 proteins (P<0.05), and higher expression levels of HO-1 and GSH-Px4 proteins (P<0.01). ConclusionGR-containing serum can reduce the inflammatory cytokines and oxidative stress levels in LPS-induced Caco2 cells. Its mechanism is related to the promotion of Nrf2/HO-1 signaling pathway expression, alleviating intracellular lipid peroxidation and inhibiting ferroptosis.
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Objective: To investigate the histological features and clinical manifestations in different types of cardiac amyloidosis to improve diagnostic accuracy. Methods: The histopathological features and clinical manifestations of 48 patients diagnosed with cardiac amyloidosis by Congo red stain and electron microscopy through endomyocardial biopsy were collected in West China Hospital of Sichuan University from January 2018 to December 2021. Immunohistochemical stains for immunoglobulin light chains (κ and λ) and transthyretin protein were carried out, and a review of literature was made. Results: The patients age ranged from 42 to 79 years (mean 56 years) and the male to female ratio was 1.1 to 1.0. The positive rate of endomyocardial biopsy was 97.9% (47/48), which was significantly higher than that of the abdominal wall fat (7/17). Congo red staining and electron microscopy were positive in 97.9% (47/48) and 93.5% (43/46), respectively. Immunohistochemical stains showed 32 cases (68.1%) were light chain type (AL-CA), including 31 cases of AL-λ type and 1 case of AL-κ type; 9 cases (19.1%) were transthyretin protein type (ATTR-CA); and 6 cases (12.8%) were not classified. There was no significant difference in the deposition pattern of amyloid between different types (P>0.05). Clinical data showed that ATTR-CA patients had less involvement of 2 or more organs and lower N-terminal pro-B-type natriuretic peptide (NT-proBNP) than the other type patients (P<0.05). The left ventricular stroke volume and right ventricular ejection fraction of ATTR-CA patients were better than the other patients (P<0.05). Follow-up data of 45 patients was obtained, and the overall mean survival time was 15.6±2.0 months. Univariate survival analysis showed that ATTR-CA patients had a better prognosis, while cardiac amyloidosis patients with higher cardiac function grade, NT-proBNP >6 000 ng/L, and troponin T >70 ng/L had a worse prognosis (P<0.05). Multivariate survival analysis showed that NT-proBNP and cardiac function grade were independent prognostic factors for cardiac amyloidosis patients. Conclusions: AL-λ is the most common type of cardiac amyloidosis in this group. Congo red staining combined with electron microscopy can significantly improve the diagnosis of cardiac amyloidosis. The clinical manifestations and prognosis of each type are different and can be classified based on immunostaining profile. However, there are still a few cases that cannot be typed; hence mass spectrometry is recommended if feasible.
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Humanos , Masculino , Femenino , Adulto , Persona de Mediana Edad , Anciano , Prealbúmina/metabolismo , Volumen Sistólico , Cardiomiopatías/patología , Rojo Congo , Función Ventricular Derecha , Amiloidosis/patología , PronósticoRESUMEN
OBJECTIVE@#To investigate the significance of anti-histidyl tRNA synthetase (Jo-1) antibody in idiopathic inflammatory myopathies (IIM) and its diseases spectrum.@*METHODS@#We enrolled all the patients who were tested positive for anti-Jo-1 antibody by immunoblotting in Peking University People's Hospital between 2016 and 2022. And the patients diagnosed with anti-synthetase antibody syndrome (ASS) with negative serum anti-Jo-1 antibody were enrolled as controls. We analyzed the basic information, clinical characteristics, and various inflammatory and immunological indicators of the patients at the onset of illness.@*RESULTS@#A total of 165 patients with positive anti-Jo-1 antibody were enrolled in this study. Among them, 80.5% were diagnosed with connective tissue disease. And 57.6% (95/165) were diagnosed with IIM, including ASS (84/165, 50.9%), immune-mediated necrotizing myopathy (7/165, 4.2%) and dermatomyositis (4/165, 2.4%). There were 23.0% (38/165) diagnosed with other connective tissue disease, mainly including rheumatoid arthritis (11/165, 6.7%), undifferentiated connective tissue disease (5/165, 3.0%), interstitial pneumonia with autoimmune features (5/165, 3.0%), undifferentiated arthritis (4/165, 2.4%), Sjögren's syndrome (3/165, 1.8%), systemic lupus erythematosus (3/165, 1.8%), systemic vasculitis (3/165, 1.8%), and so on. Other cases included 3 (1.8%) malignant tumor patients, 4 (2.4%) infectious cases and so on. The diagnoses were not clear in 9.1% (15 /165) of the cohort. In the analysis of ASS subgroups, the group with positive serum anti-Jo-1 antibody had a younger age of onset than those with negative serum anti-Jo-1 antibody (49.9 years vs. 55.0 years, P=0.026). Clinical manifestations of arthritis (60.7% vs. 33.3%, P=0.002) and myalgia (47.1% vs. 22.2%, P=0.004) were more common in the ASS patients with positive anti-Jo-1 antibody. With the increase of anti-Jo-1 antibody titer, the incidence of the manifestations of arthritis, mechanic hands, Gottron sign and Raynaud phenomenon increased, and the proportion of abnormal creatine kinase and α-hydroxybutyric dehydrogenase index increased in the ASS patients. The incidence of myalgia and myasthenia were significantly more common in this cohort when anti-Jo-1 antibody-positive ASS patients were positive for one and more myositis specific antibodies/myositis associated autoantibodies (P < 0.05).@*CONCLUSION@#The disease spectrum in patients with positive serum anti-Jo-1 antibody includes a variety of diseases, mainly ASS. And anti-Jo-1 antibody can also be found in many connective tissue diseases, malignant tumor, infection and so on.
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Humanos , Persona de Mediana Edad , Mialgia , Miositis/epidemiología , Autoanticuerpos , Enfermedades del Tejido Conjuntivo , Artritis Reumatoide , NeoplasiasRESUMEN
Vaccine-associated uveitis (VAU) usually refers to a rare adverse reaction that occurs after vaccination. The clinical manifestations of VAU are most often anterior with mild symptoms and responded promptly to topical corticosteroids. However, more severe forms of posterior and panuveitis may also occur, such as multiple evanescent white dot syndrome, Vogt-Koyanagi-Harada syndrome, and acute posterior multifocal placoid pigment epitheliopathy. The pathogenesis of VAU is still unclear. Currently, it mainly includes vaccine Shoenfeld syndrome, type Ⅲ hypersensitivity reaction caused by immune complex deposition, direct infection with live attenuated vaccine, and molecular mimicry theory. VAU is self-limiting, and most patients heal without treatment. In the future, it is recommended to ask all patients with uveitis about their recent vaccination history in the clinic. For patients with inactivated vaccine or recombinant/subunit vaccination history, the possibility of developing Shoenfeld syndrome should be considered, and the history, signs and symptoms related to autoimmune diseases should be carefully looked for.
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Objective To investigate the effects of Echinococcus multilocularis on the phenotypic transformations of glucose metabolism, polarization types and inflammatory responses in macrophages, so as to provide insights into elucidation of echinococcosis pathogenesis. Methods Bone marrow cells were isolated from C57BL/6J mice at ages of 6 to 8 weeks, and induced into bone marrow-derived macrophages (BMDMs) with mouse macrophage colony-stimulating factor (M-CSF), which served as controls (BMDMs-M0). BMDMs-M0 induced M2 macrophages by interleukin-4 for 24 hours served as the IL-4 induction group, and BMDMs-M0 co-cultured with 2.4 ng/mL E. multilocularis cystic fluid (CF) served as the BMDM-CF co-culture group, while BMDMs-M0 co-cultured with E. multilocularis protoscolex (PSC) at a ratio of 500:1 served as the BMDM-PSC co-culture group. The types of polarization of BMDMs co-cultured with E. multilocularis CF and PSC were analyzed using flow cytometry, and the expression of macrophage markers, inflammatory factors, and glucose metabolism-related enzymes was quantified using fluorescent quantitative real-time PCR (qPCR) and Western blotting assays. Results There were significant differences among the four groups in terms of Arginase-1 (Arg1) (F = 1 457.00, P < 0.000 1), macrophages-derived C-C motif chemokine 22 (Ccl22) (F = 22 203.00, P < 0.000 1), resistin-like α (Retnla) (F = 151.90, P < 0.000 1), inducible nitric oxide synthase (iNOS) (F = 107.80, P < 0.001), hexokinase (HK) (F = 9 389.00, P < 0.000 1), pyruvate kinase (PK) (F = 641.40, P < 0.001), phosphofructokinase 1 (PFK1) (F = 43.97, P < 0.01), glucokinase (GK) (F = 432.50, P < 0.000 1), pyruvate dehydrogenase kinases1 (PDK1) (F = 737.30, P < 0.000 1), lactic dehydrogenase (LDH) (F = 3 632.00, P < 0.000 1), glucose transporter 1 (GLUT1) (F = 532.40, P < 0.000 1), glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (F = 460.00, P < 0.000 1), citrate synthase (CS) (F = 5 642.00, P < 0.01), glycogen synthase1 (GYS1) (F = 273.30, P < 0.000 1), IL-6 (F = 1 823.00, P < 0.000 1), IL-10 (F = 291.70, P < 0.000 1), IL-1β (F = 986.60, P < 0.000 1), and tumor necrosis factor (TNF)-α (F = 334.80, P < 0.000 1) and transforming growth factor (TGF)-β mRNA expression (F = 163.30, P < 0.001). The proportion of M2 macrophages was significantly higher than that of M1 macrophages in the BMDM-PSC co-culture group [(22.87% ±1.48%) vs. (1.70% ±0.17%); t = 24.61, P < 0.001], and the proportion of M2 macrophages was significantly higher than that of M1 macrophages in the BMDM-CF co-culture group [(20.07% ±0.64%) vs. (1.93% ±0.25%); t = 45.73, P < 0.001]. The mRNA expression of M2 macrophages markers Arg1, Ccl22 and Retnla was significantly higher in the BMDM-CF and BMDM-PSC co-culture groups than in the control group (all P values < 0.01), and no significant difference was seen in the mRNA expression of the M1 macrophage marker iNOS among the three groups (P > 0.05), while qPCR assay quantified higher mRNA expression of key glycolytic enzymes HK, PK and PFK, as well as inflammatory factors IL-10, IL-1β, TNF-α and TGF-β in the BMDM-CF and BMDM-PSC co-culture groups than in the control group (all P values < 0.01). Western blotting assay determined higher HK, PK and PFK protein expression in the BMDM-PSC co-culture group than in the control group (all P values < 0.05), and qPCR quantified higher GLUT1, GAPDH and IL-6 mRNA expression in the BMDM-CF co-culture group than in the control group (all P values < 0.05), while higher HK, PK and PFK protein and mRNA expression (all P values < 0.01), as well as lower IL-6 and TNF-α and higher TGF-β mRNA expression (both P values < 0.05) was detected in the IL-4 induction group than in the control group. Glycolytic stress test showed no significant difference in the extracellular acidification rate (ECAR) of mouse BMDM among the control group, IL-4 induction group and BMDM-PSC co-culture group (F = 124.4, P < 0.05), and a higher ECAR was seen in the BMDM-PSC co-culture group and a lower ECAR was found in the IL-4 induction group than in the control group (both P values < 0.05). Conclusions Treatment of E. multilocularis CF or PSC mainly causes polarization of BMDM into M2 macrophages, and phenotypic transformation of glucose metabolism into high-energy and high-glycolytic metabolism, and affects inflammatory responses in BMDM.
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The development of insect appendages requires the expression of multiple genes in a strict spatial and temporal order. The odd-skipped family genes are vital transcriptional factors involved in embryonic development. The development and morphogenesis of the insect wing requires multiple transcription factors to regulate the expression of wing patterning genes at the transcriptional level. However, the function of odd-related genes in insect wing morphogenesis and development during postembryonic stages is unclear. We focused on the roles of the sister of odd and bowl (sob) gene, a member of odd-skipped family genes, during the wing morphopoiesis in Bombyx mori using the clustered regularly interspaced palindromic repeats (CRISPR)/CRISPR-associated protein 9 system and in Tribolium castaneum by RNA interference. The results showed that the wings were significantly smaller and degenerated, and wing veins were indistinct in the sob gene loss-of-function group in both B. mori and T. castaneum. Quantitative real-time polymerase chain reaction revealed that the Tcsob gene regulated the expression of wing development genes, such as the cht 7 and the vg gene. The findings suggest the importance of sob gene in insect wing morphology formation during postembryonic stages.
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Bombyx , Tribolium , Animales , Bombyx/genética , Proteínas de Insectos/genética , Morfogénesis , Tribolium/genética , Alas de AnimalesRESUMEN
In this study, black phosphorus nanosheets (BP) were prepared by the ordinary liquid phase method, and resveratrol was loaded on the BP after being modified by polyethylene glycol. The brain targeting of BP was investigated by fluorescent protein labeling, and the effects of black phosphorus on cerebral ischemia/reperfusion injury were studied by 2,3,5-triphenyltetrazolium chloride (TTC) staining, neurobehavioral evaluation, and brain edema. Protein immunoblotting analysis was used to explore the molecular mechanism of the BP drug delivery system on ischemic brain injury. Hemolysis test and hematoxylin-eosin (H&E) staining were used to evaluate its biocompatibility. The results showed that BP had excellent drug loading capacity, uniform drug loading system structure and particle size, stable drug release curve, and excellent photothermal effect. Through the analysis and comparison of fluorescence intensity, it was found that BP can increase the permeability of blood-brain barrier (BBB) under the condition of near-infrared light assisted irradiation, and make drugs more pass through the BBB. In addition, the black phosphorus nano tablet drug delivery system can significantly improve the neurobehavioral disorder of mice after modeling, and the cerebral infarction area and brain edema degree are significantly decreased. Western blot experiments showed that the drug delivery system could play an anti-ischemic brain injury role by activating the expression of antioxidant signaling pathway proteins nuclear factor E2-related factor 2 (Nrf2) and heme oxygenase-1 (HO-1). The hemolysis test and H&E test results of the BP drug carrier system showed that it had no obvious toxicity and high safety. In conclusion, the BP prepared in this study had high drug loading, good photothermal performance, and high safety. Under the near-infrared condition, they also have certain brain targeting ability, which can improve the therapeutic effect of drugs in the brain. Animal welfare and experimental procedures were following the regulations of the Animal Ethics Committee of the First Affiliated Hospital of Shihezi University.
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Objective:To observe aqueous cytomegalovirus (CMV) DNA load in patients with cytomegalovirus retinitis (CMVR) after allogeneic hematopoietic stem cell transplantation (Allo-HSCT), and to explore influencing factors for transient elevation of CMV-DNA load during the treatment.Methods:A retrospective study. From January 2016 to July 2020, 28 eyes of 19 patients with CMVR after Allo-HSCT diagnosed in the Department of Ophthalmology of Peking University People's Hospital were included in the study. Among them, there were 8 males with 12 eyes, 11 females with 16 eyes; the mean age was 28 years; 10 patients were unilateral and 9 patients were bilateral. During the course of treatment and follow-up, the blood CMV-DNA remained negative. All patients were treated with intravitreal injection of 60 mg/ml ganciclovir 0.05 ml (containing ganciclovir 3 mg), twice a week for two weeks in induction phase and weekly injection in maintenance phase. Aqueous humor sample was collected during injection of ganciclovir (IVG) and CMV-DNA load was determined by real-time quantitative polymerase chain reaction. Intravitreal treatment was terminated if aqueous CMV-DNA load turned negative after the fourth or later intravitreal injection. The patients were followed up every 2 weeks for at least 6 months. Serum CMV-DNA was negative in all patients during treatment and follow-up. All the eyes were divided into continuous decline group and non-continuous decline group depending on whether there was transient elevation of aqueous CMV-DNA load, and data between two groups were compared. Pearson linear regression analysis was used to analyze the correlation between aqueous CMV-DNA load and injection times or treatment duration.Results:At the end of treatment, the median number of IVG in the affected eye was 7 (4, 9). The results of correlation analysis showed that the aqueous humor CMV-DNA load of the affected eye was related to the number of treatments [ R2=0.385, P<0.000 1, B=-0.237 log 10 copies/(ml·time)], and the duration of treatment [ R2=0.394, P <0.000 1, B=-0.301 log 10 copies/(ml·week)] were negatively correlated. Among the 28 eyes, 13 eyes (46.4%, 13/28) in the continuous decline group and 15 eyes (53.6%, 15/28) in the non-sustained decline group. Baseline visual acuity ( t=-1.223), intraocular pressure ( t=1.538), aqueous humor CMV-DNA load ( t=-0.109), retinitis lesion area ( Z=-0.308) in the continuous decline group and the non-continuous decline group), the number of quadrants involved ( Z=-0.024) and whether the macula was involved ( Z=-1.826), combined with anterior segment inflammation ( Z =-0.499), combined with high intraocular pressure ( Z=-1.342), terminal visual acuity ( t =-0.845), intraocular pressure ( t=-0.068), total IVG times ( Z=0.907), age ( Z=-0.832), gender composition ( Z=-1.074), etc. The difference was not statistically significant ( P>0.05). Conclusion:The CMV-DNA load in aqueous humor decreases by about 50% every week during the treatment of CMVR eyes after Allo-HSCT; the transient increase in the CMV-DNA load in the aqueous humor during treatment does not affect the treatment process and clinical prognosis.
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Objective To investigate the changes of mitochondrial metabolic functions of macrophages following Echinococcus multilocularis infections, so as to provide insights into the pathogenesis of alveolar echinococcosis. Methods Two groups were assigned according to different treatment methods. In the culture group, mouse leukemic monocyte macrophage RAW264.7 cells were cultured with 2 000 E. multilocularis at a ratio of 500∶1, while RAW264.7 cells in the control group were given no treatment. Then, both the culture and control groups were further divided into the 24 h and 72 h subgroups. Mitochondria were stained with MitoTracker® Deep Red FM and the mean fluorescence intensity of macrophage mitochondria was measured with the Cytation 5 Cell Imaging Multi-Mode Reader. The mitochondrial DNA copy number was quantified using the quantitative real-time PCR (qPCR) assay, and the mitochondrial energy metabolism was monitored using the Seahorse XF assay. In addition, the mitochondrial reactive oxygen species and mitochondrial membrane potential were detected using flow cytometry. Results The mean fluorescence intensities of macrophage mitochondria were significantly lower in the 24 h (15 341 ± 2 532 vs. 17 823 ± 3 429; t = 6.379, P < 0.01) and 72 h (18 102 ± 3 505 vs. 21 511 ± 5 144; t = 17.680, P < 0.01) culture subgroups than in the corresponding control subgroups, and lower mitochondrial DNA copy numbers were measured in the 72 h culture subgroup than in the 72 h control group [(3.23 × 109 ± 1.78 × 107) vs. (4.39 × 109 ± 3.70 × 107); t = 8.85, P < 0.001]. The oxygen consumption rates were significantly greater in the 24 h [(241.70 ± 73.13) pmol/min vs. (69.05 ± 52.30) pmol/min; t = 7.89, P < 0.01] and 48 h culture groups [(249.50 ± 42.06) pmol/min vs. (60.28 ± 40.66) pmol/min; t = 8.64, P < 0.01] than in the corresponding control groups, and a higher extracellular acidification rate was seen in the 48 h culture group than in the 48 h control group ([ 111.6 ± 17.49) mpH/min vs. (35.05 ± 7.57) mpH/min; t = 16.90, P < 0.01]. In addition, flow cytometry detected higher mean fluorescence intensity of mitochondrial reactive oxygen species (58 264 ± 10 087 vs. 4 307 ± 97; t = 12.930, P < 0.01) and lower mitochondrial membrane potential (9.833% ± 2.285% vs. 2.667% ± 0.208%; t = 6.645, P < 0.01) in the 72 h culture group than in the control group. Conclusions E. multilocularis infection may impair mitochondrial functions and inhibit oxidative phosphorylation of macrophages, resulting in increased macrophage glycolysis. It is speculated that the alteration of macrophage metabolic states may contribute to the mechanisms underlying the development and progression of alveolar echinococcosis.
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Objective:To establish the grade evaluation method for Codonopsis Radix slices based relative quality constants, in order to provide scientific theoretical basis for grading of Codonopsis Radix slices. Method:Through literature and market research,the main production areas of Codonopsis Radix slices were determined,and 67 batches of Ludangshen slices(52 batches) and Baitiaodang slices (15 batches) were collected. The appearance traits (average quality and average thickness of Codonopsis Radix slices) were observed and measured. According to Chinese Pharmacopoeia (2015 edition), the extract and the content of Codonopsis pilosula polysaccharide was determined by phenol-sulphoacid method. Then the relative quality constant was calculated,and the results of grade evaluation were evaluated through systematic cluster analysis and correlation analysis. Result:Relative quality constants of 67 batches of Codonopsis Radix slices were between 0.32-2.97. If these samples were divided into three grades:the first-grade relative quality constants were greater than or equal to 2.08,the second grade was greater than or equal to 0.89 but less than 2.08,while the third grade was less than 0.89. The results of systematic cluster analysis showed that 67 batches of Codonopsis Radix slices were clustered into 3 categories,and the results were basically consistent with the classification. The correlation analysis showed a positive correlation between the content of the extracts and the polysaccharide content (P<0.05). Conclusion:This method links the extrinsic characteristics to the intrinsic quality,and objectively grade Codonopsis Radix slices, so as to provide a basis for its grade standards.
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Objective:To explore the protective effect and the mechanism of Danggui Shaoyaosan(DSS) on angiotensin Ⅱ (AngⅡ)/transient receptor potential cation channel 6 (TRPC6) pathway in nephrotic syndrome (NS) rats. Method:In animal experiments, doxorubicin (4 mg·kg<sup>-1</sup> for the 1<sup>st</sup> week and 2 mg·kg<sup>-1</sup> for the 2<sup>nd</sup> week) was injected twice to the tail vein of rats to induce NS model in 160 rats, which were then randomly divided into model group (normal saline), losartan group (30 mg·kg<sup>-1</sup>·d<sup>-1</sup>), and low-(4.3 g·kg<sup>-1</sup>·d<sup>-1</sup>), medium-(8.6 g·kg<sup>-1</sup>·d<sup>-1</sup>), and high-dose (17.2 g·kg<sup>-1</sup>·d<sup>-1</sup>) DSS groups. Besides, a normal group was also set. After intervention for four weeks, ultrastructure changes of the kidney were identified by transmission electron microscopy (TEM). The 24-hour urine protein was detected by kits. Radioimmunoassay was used to detect the content of AngⅡ and Calcineurin (CaN) in plasma. Western blot was used to detect the protein expression of TRPC6, angiotensin Ⅱ type 1 receptor (AT1R), podocyte slit diaphragm-specific protein (Nephrin), and cysteine-aspartic acid protease-3 (Caspase-3) in the renal cortex. Immunohistochemistry was used to detect the expression of TRPC6 and AT1R in the slit diaphragm. In cell experiments, AngⅡ stimulated MPC5 podocytes. The cells were randomly divided into a normal group, an AngⅡ group, an AngⅡ+SAR7334 (TRPC6-specific inhibitor) group, an AngⅡ+5%DSS group, an AngⅡ+10%DSS group, and an AngⅡ+15%DSS group. Western blot was used to detect the protein expression of TRPC6, AT1R, Nephrin, and Caspase-3 in podocytes. Result:Compared with the normal group, the model group showed increased 24-hour urine protein content (<italic>P</italic><0.01) and AngⅡ and CaN in plasma (<italic>P</italic><0.01), incomplete glomerular structure, the extensive fusion of podocyte process with elevated fusion rate (<italic>P</italic><0.01), increased expression distribution of AT1R and TRPC6 in the renal cortex, and up-regulated protein expression of AT1R, TRPC6, and Caspase-3 in renal tissues (<italic>P</italic><0.01), and reduced Nephrin protein expression (<italic>P</italic><0.01). Compared with model group, the losartan group and the high-dose DSS group exhibited decreased 24-hour urine protein content (<italic>P</italic><0.01) and the content of AngⅡ and CaN in plasma (<italic>P</italic><0.01), improved glomerular structure, reduced fusion rate of podocyte process (<italic>P</italic><0.01), diminished expression distribution of TRPC6 and AT1R in the renal cortex, declining protein expression of AT1R, TRPC6 and Caspase-3 in renal tissues (<italic>P</italic><0.01), and elevated Nephrin protein expression (<italic>P</italic><0.01). Additionally, compared with the normal podocytes, AngⅡ-stimulated podocytes showed increased protein expression of AT1R, TRPC6 and Caspase-3 (<italic>P</italic><0.01), and decreased expression of Nephrin (<italic>P</italic><0.01). Compared with the AngⅡ group, the AngⅡ+SAR7334 group displayed reduced protein expression of AT1R, TRPC6, and Caspase-3 (<italic>P</italic><0.01) and increased protein expression of Nephrin (<italic>P</italic><0.01). Conclusion:DSS can improve the pathological characteristics of NS presumedly by inhibiting the interaction between AngⅡ and TRPC6 in podocytes and improving the structural integrity of podocytes to repair the damage of glomerular molecular barrier and slow down the progression of NS-induced proteinuria.
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Objective:To explore the diagnostic value of serum cytokines and autoantibodies in children with central diabetes insipidus.Methods:A retrospective study was used.A total of 52 children diagnosed with central diabetes insipidus who were hospitalized in Qingdao municipal hospital affiliated to Qingdao Universityfrom January 2016 to January 2020 were selected as the study group, and 60 healthy children with similar gender and age were selected as the control group.The general data of the study group and the control group, 24 h urine volume, urine specific gravity and urine osmotic pressure before the start of water deprivation test, the results of pituitary magnetic resonance imaging (MRI), and the concentration of serum cytokines and autoimmune antibodies were compared to evaluate the value of serum cytokines and autoimmune antibodies in the prediction of central diabetes insipidus.Results:The 24 h urine volume in the study group (201.6 ± 9.9 mL/kg)was significantly higher than that in the control group(9.06±12.5 mL/kg). The urine specific gravity(1.006±0.002) and urine osmotic pressure((116.4±23.5) mOsm/L)were significantly lower than those in the control group((1.013±0.004) and (649.6±72.4) mOsm/L)( t=15.378, 13.298, 45.625, all P<0.001). MRI examination showed that the proportion of disappearance of high signal in posterior pituitary (46.1%(24/52)), the proportion of thickening of the pituitary stalk(32.7%(17/52))and abnormal signal in pituitary(7.7%(4/52)) in study group were significantly higher than the control group(0%(0/0)) (all P<0.05); the levels of serum IL-6, TNF-α, IFN-γ, IL-1β ((117.0±7.8), (234±21.4), (178.6±15.1), (87±6.5) ng/L)and autoimmune antibodies ((654.1±194.3) ng/L) in study group were significantly increased compared to the control((12.5±2.3), (32.5±9.7), (37.8±7.3), (22.6±4.7), (98.0±23.7) ng/L) ( t value were 17.031, 27.490, 15.670, 12.360 and 109.984, respectively; all P<0.05). Conclusion:The concentration of serum cytokines and autoimmune antibodies has a significant clinical value in the evaluation of central diabetes insipidus in children.Combined with head MRI examination, it has a certain value in the diagnosis, treatment and prognosis of central diabetes insipidus in children.
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Cytomegalovirus (CMV) retinitis (CMVR) is a common opportunistic infection of the eye after allogeneic hematopoietic stem cell transplantation in patients with hematological diseases. It often occurs within 3 months after the operation, with CMV activation and high blood CMV peaks. It often occurs on patients with long-term CMV viremia, human leukocyte antigen incompatible transplantation, unrelated donor transplantation, haploid transplantation, childhood hematopoietic stem cell transplantation, delayed lymphocyte engraftment, acute and chronic graft-versus-host disease after surgery. The visual prognosis of patients is related to the area of CMVR lesions on the retina, the number of quadrants involved, whether the macula is involved, and the CMV load of the vitreous body is involved, and it is not related to whether the Epstein-Barr virus infection is combined with blood and vitreous humor. The incidence of CMVR is increasing year by year. It is helpful that paying attention to systemic risk factors and epidemiology can provide more effective guidance for ophthalmologists during diagnosis and treatment, help patients improve the prognosis of vision, and reduce or even avoid the occurrence of blindness caused by CMVR.
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The aliasing effect in the discrete Fourier transform inherent will impose a serious detrimental effect on conventional phase retrieval measurement accuracy with under-sampled intensity. In this Letter, we describe a modal-based nonlinear optimization phase retrieval approach that is capable of retrieving wavefront measurements using under-sampled intensities. The extended Nijboer-Zernike theory is introduced to establish an analytic solution between wavefront phase and intensity image, and then nonlinear optimization is further utilized to solve wavefront aberration coefficients from under-sampled intensity data. The feasibility and accuracy of the algorithm are verified by simulations and experiments. This is a promising method that is especially suitable for full field phase recovery of optical systems with a relatively high numerical aperture.
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Lymphoma is common in IgG4-related disease, but diffuse large B-cell lymphoma in IgG4-related ophthalmic disease is rare, especially involving the same organ affected by IgG4-related ophthalmic disease. The authors report a 44-year-old man presented with bilateral proptosis for 10 years and rapid loss of vision in the right eye for the last 1 month. The authors performed orbital exenteration for the right eye and resected the tumor in the left eye. Based on the clinicopathologic findings, tumors of the right and left eyes were diagnosed as diffuse large B-cell lymphoma and IgG4-related ophthalmic disease, respectively. This case highlights the risk of malignant transformation of IgG4-related ophthalmic disease.
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Oftalmopatías , Enfermedad Relacionada con Inmunoglobulina G4 , Linfoma de Células B Grandes Difuso , Adulto , Humanos , Inmunoglobulina G , Enfermedad Relacionada con Inmunoglobulina G4/diagnóstico , Linfoma de Células B Grandes Difuso/diagnóstico , MasculinoRESUMEN
Objective:To explore the clinical significance of serum nuclear factor κ B, interleukin(IL)-4, IL-10, IL-12, interferon(IFN)- γ expression in subclinical hypothyroidism with gestational diabetes mellitus.Methods:Thirty pregnant women with subclinical hypothyroidism combined with gestational diabetes mellitus in Tangshan Maternal and Child Health Hospital from January 2017 to October 2018 were retrospectively analyzed as group A. Thirty three pregnant women with subclinical hypothyroidism were selected as group B, 35 pregnant women with gestational diabetes mellitus as group C and 40 healthy pregnant women as control group.ELISA was used to detect NF-κB, IL-4, IL-10, IL-12 and IFN-γ, and the results were analyzed and compared.Results:The serum levels of NF-κB were (15.91±5.68), (13.22±5.23), (12.97±5.11), (9.74±3.85) μg/L, IL-12 were (28.91±6.84), (21.64±5.72), (22.23±5.91), (13.68±3.76) ng/L, and serum IFN-γ levels were (23.74±5.55), (18.26±4.63), (17.85±4.31), (12.69±3.85) ng/L in A, B, C and the control group respectively.There were statistically significant differences in the three indicators between groups ( F=5.118, 6.821, 7.133, all P<0.05), and group A was higher than group B, C and control, the differences were statistically significant (all P<0.05); the levels of serum IL-4 in group A, B, C and control group were (8.91±3.99), (10.84±4.47), (11.27±4.62), (13.68±5.46) ng/L, respectively.The levels of serum IL-10 were (10.91±3.86), (13.05±4.58), (12.83±4.69), (15.82±5.33) ng/L, respectively.The differences of serum IL-4 and IL-10 between groups were statistically significant ( F=5.075, 5.616, all P<0.05), and serum IL-4 and IL-10 in group A were lower than those in group B, group C and control group.The levels of serum IL-4 and IL-10 in group B and C were lower than those in control group (all P<0.05). Conclusion:The activation of NF-kB signaling pathway and its related cytokines may be the influencing factors for the development of subclinical hypothyroidism with gestational diabetes mellitus.
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.This study aimed to investigate the hypolipidemic and antioxidant activities of volatile oils from Michelia martini Levl. The antioxidant property of volatile oils from Michelia martini in vitro was investigated by establishment of various systems. High fat diet induced rats were used to assess the hypolipidemic and antioxidant activities of Michelia martini volatile oils in vivo. The level of total cholesterol, triglycerides, high density lipoprotein cholesterol, low density lipoprotein cholesterol, alanine transaminase, aspartate aminotransferase, alkaline phosphatase and gamma-glutamyl transpeptidase in serum, and the activities of catalase, malondialdehyde, super oxide dismutase and glutathione in liver of rats were assayed by standard procedures. Our results showed that Michelia martini exhibits strong hypolipidemic and antioxidant activities both in vitro and vivo. Our data were also supplemented with histopathological studies on liver tissues and aorta sections of rats.
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Antioxidantes/farmacología , Hipolipemiantes/farmacología , Magnoliaceae/química , Aceites Volátiles/farmacología , Aceites de Plantas/farmacología , Anfotericina B/toxicidad , Animales , Antioxidantes/aislamiento & purificación , Compuestos de Bifenilo/química , Células CHO , Cricetulus , Dieta Alta en Grasa/efectos adversos , Relación Dosis-Respuesta a Droga , Hiperlipidemias/sangre , Hiperlipidemias/prevención & control , Hipolipemiantes/aislamiento & purificación , Lípidos/sangre , Masculino , Aceites Volátiles/aislamiento & purificación , Picratos/química , Hojas de la Planta/química , Aceites de Plantas/aislamiento & purificación , Ratas Sprague-DawleyRESUMEN
Objective To investigate the efficacy of plasma exchange therapy in the treatment of rhabdomyolysis syndrome (RMS) caused by wasp stings. Methods Sixty patients with RMS caused by wasp stings were enrolled in the Poisoning Treatment Ward of the Affiliated Hospital of Southwest Medical University from January 2015 to June 2019, including 40 patients complicated with acute kidney injury (AKI). After admission, the patients were given local treatment and systemic medication (intravenous infusion of glucocorticoid, calcium gluconate injection, intramuscular injection of isopropazine hydrochloride and other antihistamines). Plasma exchange was performed on the first and second day after admission, and according to the renal function situations, the patients with AKI underwent intermittent hemodialysis. The changes of urea (Ur), creatinine (Cr), hypersensitive troponin T (hs-TnT), myoglobin (MYO), creatine kinase isoenzyme (CK-MB), lactic dehydrogenase (LDH) and pathological changes of kidney were monitored on admission and after twice of plasma exchanges, and the above results of indexes were compared and analyzed. Results With the prolongation of the plasma exchange time, after the first plasma exchange the level of hs-TnT was significantly higher than that before the plasma exchange [μg/L: 1.30 (0.16, 4.37) vs. 0.26 (0.06, 1.26)], and the levels of LDH and CK-MB were lower than those before the exchange [LDH (μmol·s-1·L-1): 14.01 (6.73, 31.52) vs. 20.55 (8.73, 42.46), CK-MB (U/L): 41.25 (21.27, 102.83) vs. 89.92 (35.85, 163.53), both P < 0.05]; after the second plasma exchange, the Ur, Cr, LDH, hs-TnT, MYO and CK-MB were decreased significantly compared with those before the exchange [Ur (mmol/L): 9.77 (6.43, 11.90) vs. 11.58 (7.65, 19.49), Cr (μmol/L): 90.35 (67.10, 336.10) vs. 115.25 (77.50, 288.83), LDH (μmol·s-1·L-1): 7.84 (5.14, 18.68) vs 20.55 (8.73, 42.46), hs-TnT (μg/L): 0.02 (0.01, 0.09) vs. 0.26 (0.06, 1.26), MYO (μg/L): 200.00 (70.39, 1 000.00) vs. >1 000.00 (1 000.00, >1 000.00), CK-MB (U/L): 4.34 (1.86, 23.65) vs. 89.92 (35.85, 163.56), all P < 0.05]. Under light microscope, renal tubular epithelial cells fell off and showed"naked membrane", and infiltration of a small amount of inflammatory cells could be seen. Conclusion Plasma exchange can effectively remove macromolecules such as MYO, hs-TnT, CK-MB and LDH, reduce acute renal damage caused by rhabdomyolysis syndrome, shorten the course of disease and decrease mortality.