AIM: To investigate the effects of intracellular free calcium ([Ca 2+ ]i) from different resources on the proliferation mediated by mitogen activated protein kinase (MAPK) in vascular smooth muscle cells (VSMCs). METHODS: Cultured VSMCs were used in all experiments. Calcium influx was stimulated by angiotension Ⅱ(AngⅡ). The release of intracellular calcium stores was induced by inositol trisphosphate(IP 3) and ryanodine(RY). MAPK activity was measured by [?- 32 P]-ATP incorporation , MAPK protein expression by western blot, VSMCs proliferation by -Leucine(-Leu) and -Thymidine(-TdR) incorporation. RESULTS: Compared to the control VSMCs, AngⅡ, IP 3 and RY significantly increased [Ca 2+ ]i concentration , activity of MAPK and its protein content in VSMCs. The promotion of -Leu and -TdR incorporation in VSMCs was also observed ( P

OBJECTIVETo explore the mechanism and strategies of prevention and treatment of vision deterioration after transsphenoidal surgery for removal of pituitary adenoma.

METHODSFrom January 1980 to February 2001, 1 412 patients were operated on by transsphenoidal surgery to remove pituitary adenoma. Fourteen patients experienced vision deterioration after operation. Clinical data from the 14 patients with vision deterioration were analyzed retrospectively.

RESULTSIn this group, the incidence of vision deterioration was 0.99%. Over packing of the adenoma bed was seen in 5 patients, apoplexy of residual tumor in 3, high intracranial pressure in 2, vascular spasm in 2, injury of the optic nerve in the cannel in 1 and unknown reason in 1.

CONCLUSIONSVision deterioration is a severe complication after transsphenoidal surgery. Early diagnosis and treatment can greatly improve the vision.

Adenoma ; surgery ; Adult ; Aged ; Female ; Humans ; Ischemia ; complications ; Male ; Middle Aged ; Optic Nerve ; blood supply ; Optic Nerve Injuries ; etiology ; prevention & control ; Pituitary Neoplasms ; surgery ; Postoperative Complications ; etiology ; prevention & control ; Retrospective Studies ; Sphenoid Sinus ; Vision Disorders ; etiology ; prevention & control

Objective To study the therapeutic effect of anti-CD80 bivalent antibody on mouse lu-pus nephritis and to explore the possible molecular mechanism. Methods A mouse model of lupus nephritis was established through intraperitoneal injection of 0. 5 ml of pristine in female C57BL/6J mice. Appearance of urinary protein and significantly increased levels of peripheral antinuclear antibody ( ANA) and anti-doub-le-stranded DNA ( anti-dsDNA) antibody in the fourth month after injection indicated that the mouse model was established successfully. Then the mice were divided into two groups including anti-CD80 bivalent anti-body intervention group (injected with 200μg of anti-CD80 bivalent antibody at day 1, 3, 5, 8 and 15, fol-lowed by three times of injection with an interval of one month) and model group ( injected with the same protein using the same strategy). A normal control group was set up accordingly. Albustix test paper was used to monitor the dynamic changes in mouse urinary protein. Flow cytometry was used to analyze the acti-vation of immune-related cells in spleen. Levels of autoantibodies ( ANA and anti-dsDNA) and levels of IFN-γ and IL-4 in serum were detected by indirect immunofluorescence assay. Renal tissue samples were an-alyzed with hematoxylin and eosin ( HE) staining and immunocomplex ( IC) assay. Results Urinary pro-tein level of the anti-CD80 bivalent antibody intervention group was significantly lower than that of the model group (P<0. 05). Activated macrophages, dendritic cells, neutrophils and B cells in spleen tissues of the anti-CD80 bivalent antibody intervention group were significantly less than those of the model group ( P<0. 05), and the numbers of CD4+ and CD154+ T cells were significantly less than those of the model group (P<0. 05). Positive rates and titers of ANA and dsDNA in serum samples of the intervention group were lower than those of the model group (P<0. 05). Levels of IFN-γand IL-4 in serum samples of the interven-tion group were decreased as compared with those of the model group (P<0. 05). HE staining and immuno-fluorescence assay showed that glomerular inflammatory injury and necrosis were alleviated and kidney im-mune complex deposition was reduced after anti-CD80 bivalent antibody intervention. Conclusion Anti-CD80 bivalent antibody specifically binds to the CD80 molecule on antigen presenting cell surface, blocks the CD80/CD28 co-stimulatory signaling pathway and down-regulates the body′s immune response, which al-leviates and reverses the lupus-like nephritis-induced pathological damages in mice.

Objective:To investigate the effect of curcumin on renal fibrosis in lipopolysaccharide (LPS) induced acute respiratory distress syndrome (ARDS) in adult SD rats.Methods:Twenty-four male SD rats were randomly (random number) divided into four groups: control group, ARDS group, low dose group, and high dose group ( n=6 per group). In the control group, the rats were given atomization intratracheal of standard saline 2 mL/kg; in the ARDS group, low-dose group, and high-dose group, the rats were given atomization intratracheal of 4 mg/kg LPS; in the low-dose group, the rats were given curcumin 100 mg/d by the oral administration, and in the high-dose group, the rats were given curcumin or 200 mg/d respectively. After seven days, the rats were sacrificed. The superoxide dismutase (SOD) activity and the content of malondialdehyde (MDA) and glutathione (GSH) in renal tissue were detected by colorimetric assay. Nuclear factor kappa-B p65 (NF-κB p65) and transforming growth factor-β1 (TGF-β1) were detected by Western blot. The expression of interleukin-6 (IL-6) mRNA, proline hydroxylase 3 (PHD3) mRNA, vascular endothelial growth factor (VEGF) mRNA and erythropoietin receptor (EPOR) mRNA were detected by quantitative real-time PCR (qRT-PCR). HE staining and Masson staining were used to assess pathological damage. One-way analysis of variance was used for comparison among multiple groups and SNK method was used for comparison between two groups. Results:Compared with the control group, the SOD activity and GSH content in the ARDS group, low-dose group, and high-dose group were significantly decreased (all P<0.05); the protein expression levels of MDA, NF-κB p65, and TGF-β1 were increased significantly, and IL-6 mRNA, PHD3 mRNA, VEGF mRNA, and EPOR mRNA were significantly upregulated (all P<0.05). HE staining showed inflammatory cell infiltration, and fibrogenesis in kidney tissue, and Masson staining showed deposition of collangen-like substance. Compared with the ARDS group, SOD activity and GSH content were increased, while the protein expression of NF-κB p65 and TGF-β1, IL-6 mRNA, PHD3 mRNA, VEGF mRNA, and EPOR mRNA were decreased significantly in the low-dose group and high-dose group (all P<0.05). Curcumin therapy reduced inflammatory cellular infiltration, and the deposition of collagen-like substance in kidney tissue. Compared with the low-dose group, SOD activity and GSH content were increased in the high-dose group (all P<0.05), and the protein expression of NF-κB p65 and TGF-β1, IL-6 mRNA, PHD3 mRNA, VEGF mRNA, and EPOR mRNA were decreased significantly in the high-dose group (all P<0.05). The high-dose group exhibited a significant reduction in edema, and a decrease of the deposition of collagen-like substance in kidney tissue. Conclusions:Curcumin can inhibit the development of renal fibrosis induced by acute respiratory distress syndrome in rats, and its mechanism may be related to inhibiting the expression of inflammatory factors and enhancing hypoxia tolerance.

Objective To investigate the impact of dexamethasone(DEX)on diaphragmatic atrophy caused by acute respiratory distress syndrome(ARDS)and its correlation with diaphragmatic protein metabolism.Methods Twenty healthy male Sprague-Dawley(SD)rats were randomly assigned to control,ARDS model,low-dose DEX,and high-dose DEX group,with each group consisting of five rats.ARDS was induced in the rats by intratracheal administration of lipopolysaccharide(LPS)at 4 mg/kg.Conversely,intratracheal saline was administered to the control group at 2 mL/kg.Following the induction of the model,an intraperitoneal injection of DEX at 1 mg·kg-1·d-1 was administered to the low-dose DEX group.Conversely,DEX at 5 mg·kg-1·d-1 was administered to the high-dose group for 7 consecutive days.Subsequently,on the eighth day of the experiment,the diaphragmatic weight of all rats was measured.Real-time quantitative polymerase chain reaction(PCR)was utilized to assess the mRNA expression of interleukins(IL-1β,IL-18)in each group.Western blotting was employed to determine the protein expression levels of nuclear factor-κB(NF-κB)p65,NOD-like receptor protein 3(NLRP3),caspase-1,Gasdermin D(GSDMD),myosin heavy chain 2(Myh2),and F-box protein 32(Fbxo32).Additionally,immunohistochemistry was utilized to evaluate the ratio of fast to slow muscle fibers in the diaphragm.Results The ARDS model group showed significant reductions in body weight,diaphragm weight,fast muscle fibers,and Myh2 protein expression compared to the control group[body weight(g):266±17 vs.292±15,diaphragm weight(g):0.77±0.02 vs.0.92±0.08,fast muscle fibers:(74±1)%vs.(78±3)%,Myh2 protein expression(Avalue):0.75±0.07 vs.0.95±0.05,all P<0.05].Conversely,significant increases were observed in the expressions of IL-1β and IL-18 mRNA,slow muscle fibers,and the proteins NF-κB p65,NLRP3,caspase-1,GSDMD,Fbxo32[IL-1β mRNA(IL-1β/GAPDH):2.2±0.3 vs.1.0±0.2,IL-18 mRNA(IL-18/GAPDH):2.3±0.3 vs.1.0±0.3,slow muscle fibers:(26±1)%vs.(22±3)%,NF-κB p65 protein expression(A value):0.40±0.15 vs.0.17±0.05,NLRP3 protein expression(A value):0.51±0.05 vs.0.27±0.08,caspase-1 protein expression(A value):0.54±0.12 vs.0.30±0.19,GSDMD protein expression(A value):0.40±0.12 vs.0.20±0.05,Fbxo32 protein expression(A value):0.51±0.15 vs.0.33±0.08,all P<0.05].Compared with the ARDS group,both low and high doses of DEX were found to further reduce body weight,diaphragm weight,fast muscle fibers,and Myh2 protein expression,and further increase the expressions of IL-1β and IL-18 mRNA,slow muscle fibers,and the proteins NF-κB p65,NLRP3,caspase-1,GSDMD,Fbxo32,with the changes in the high dose DEX group being more significant than those in the low dose group[body weight(g):198±14 vs.222±16,diaphragm weight(g):0.57±0.04 vs.0.68±0.04,fast muscle fibers:(56±5)%vs.(69±2)%,Myh2 protein expression(A value):0.29±0.16 vs.0.57±0.15,IL-1βmRNA expression:5.6±1.4 vs.3.3±0.6,IL-18 mRNA expression(IL-18/GAPDH):5.8±1.2 vs.3.9±0.6,slow muscle fibers:(44±5)%vs.(31±2)%,NF-κB p65 protein expression(A value):0.87±0.04 vs.0.70±0.07,NLRP3 protein expression(A value):0.75±0.08 vs.0.63±0.04,caspase-1 protein expression(A value):0.99±0.06 vs.0.82±0.08,GSDMD protein expression(Avalue):0.85±0.11 vs.0.61±0.10,Fbxo32 protein expression(Avalue):1.00±0.10 vs.0.78±0.12,all P<0.05].Normal muscle fiber structure was revealed by microscopic observation in the control group,clear fiber separation in the ARDS model group,and disordered muscle fiber arrangement with structural distortion was noted in both low and high-dose DEX groups.Conclusion Prolonged administration of DEX may worsen diaphragmatic atrophy induced by ARDS,possibly by promoting the activation of the NLRP3 inflammasome and cell pyroptosis.

OBJECTIVE To investigate the monitoring of tacrolimus blood concentration in patients with nephrotic syndrome （NS），and to establish a prediction model for tacrolimus blood concentration. METHODS Data from 509 concentration monitoring sessions of 166 NS patients using tacrolimus were collected from January 1， 2020 to August 31， 2023 in Zhongshan Hospital Affiliated to Xiamen University. The relationship of efficacy and adverse drug reaction（ADR） with blood concentration was analyzed. A multilayer perceptron （MLP） prediction model was established by using the blood concentration monitoring data of 302 times from 109 NS patients with genetic information， and then verified. RESULTS In terms of efficacy， the median blood concentration of tacrolimus in the non-remission group was 2.20 ng/mL， which was significantly lower than that in the partial remission group （4.00 ng/mL， P＜0.001） and the complete remission group （3.60 ng/mL， P＝0.002）. In terms of ADR， the median blood concentration of tacrolimus in the ADR group was 5.01 ng/mL， which was significantly higher than that in the non-ADR group （3.37 ng/mL） （P＝0.001）. According to the subgroup analysis of the receiver operating characteristic curve， when the blood concentration of tacrolimus was ≥6.65 ng/mL， patients were more likely to develop elevated blood creatinine [area under the curve （AUC） was 0.764， P＜0.001）； when the blood concentration of tacrolimus was ≥6.55 ng/mL， patients were more likely to develop blood glucose （AUC＝0.615， P＝ 0.005）. The established MLP prediction model has a loss function of 0.9， with an average absolute error of 0.279 5 ng/mL between the predicted and measured values. The determination coefficient of the validation scatter plot was 0.984， indicating an excellent predictive performance of the model. CONCLUSION Tacrolimus blood concentration has an impact on both efficacy and ADR in NS patients. The use of the MLP model for predicting blood concentration exhibits high accuracy with minimal error between predicted and measured values. The model can be used as an important tool in clinical individualized medication regimens.

OBJECTIVE: To evaluate the effect of curcumin on renal mitochondrial oxidative stress, nuclear factor-κB/NOD-like receptor protein 3 (NF-κB/NLRP3) inflammatory body signaling pathway and tissue cell injury in rats with acute respiratory distress syndrome (ARDS). METHODS: A total of 24 specific pathogen free (SPF)-grade healthy male Sprague-Dawley (SD) rats were randomly divided into control group, ARDS model group, and low-dose and high-dose curcumin groups, with 6 rats in each group. The ARDS rat model was reproduced by intratracheal administration of lipopolysaccharide (LPS) at 4 mg/kg via aerosol inhalation. The control group was given 2 mL/kg of normal saline. The low-dose and high-dose curcumin groups were administered 100 mg/kg or 200 mg/kg curcumin by gavage 24 hours after model reproduction, once a day. The control group and ARDS model group were given an equivalent amount of normal saline. After 7 days, blood samples were collected from the inferior vena cava, and the levels of neutrophil gelatinase-associated lipocalin (NGAL) in serum were determined by enzyme-linked immunosorbent assay (ELISA). The rats were sacrificed, and kidney tissues were collected. Reactive oxygen species (ROS) levels were determined by ELISA, superoxide dismutase (SOD) activity was detected using the xanthine oxidase method, and malondialdehyde (MDA) levels were determined by colorimetric method. The protein expressions of hypoxia-inducible factor-1α (HIF-1α), caspase-3, NF-κB p65, and Toll-like receptor 4 (TLR4) were detected by Western blotting. The mRNA expressions of HIF-1α, NLRP3, and interleukin-1β (IL-1β) were detected by reverse transcription-polymerase chain reaction (RT-PCR). Renal cell apoptosis was detected by TdT-mediated dUTP nick end labeling (TUNEL). The morphological changes in renal tubular epithelial cells and mitochondria were observed under a transmission electron microscope. RESULTS: Compared with the control group, the ARDS model group exhibited kidney oxidative stress and inflammatory response, significantly elevated serum levels of kidney injury biomarker NGAL, activated NF-κB/NLRP3 inflammasome signaling pathway, increased kidney tissue cell apoptosis rate, and renal tubular epithelial cell damage and mitochondrial integrity destruction under transmission electron microscopy, indicating successful induction of kidney injury. Following curcumin intervention, the injury to renal tubular epithelial cells and mitochondria in the rats was significantly mitigated, along with a noticeable reduction in oxidative stress, inhibition of the NF-κB/NLRP3 inflammasome signaling pathway, and a significant decrease in kidney tissue cell apoptosis rate, demonstrating a certain dose-dependency. Compared with the ARDS model group, the high-dose curcumin group exhibited significantly reduced serum NGAL levels and kidney tissue MDA and ROS levels [NGAL (μg/L): 13.8±1.7 vs. 29.6±2.7, MDA (nmol/g): 115±18 vs. 300±47, ROS (kU/L): 75±19 vs. 260±15, all P < 0.05], significantly down-regulated protein expressions of HIF-1α, caspase-3, NF-κB p65, and TLR4 in the kidney tissue [HIF-1α protein (HIF-1α/β-actin): 0.515±0.064 vs. 0.888±0.055, caspase-3 protein (caspase-3/β-actin): 0.549±0.105 vs. 0.958±0.054, NF-κB p65 protein (NF-κB p65/β-actin): 0.428±0.166 vs. 0.900±0.059, TLR4 protein (TLR4/β-actin): 0.683±0.048 vs. 1.093±0.097, all P < 0.05], and significantly down-regulated mRNA expressions of HIF-1α, NLRP3, and IL-1β [HIF-1α mRNA (2^-ΔΔCt): 2.90±0.39 vs. 9.49±1.87, NLRP3 mRNA (2^-ΔΔCt): 2.07±0.21 vs. 6.13±1.32, IL-1β mRNA (2^-ΔΔCt): 1.43±0.24 vs. 3.95±0.51, all P < 0.05], and significantly decreased kidney tissue cell apoptosis rate [(4.36±0.92)% vs. (27.75±8.31)%, P < 0.05], and significantly increased SOD activity (kU/g: 648±34 vs. 430±47, P < 0.05). CONCLUSIONS Curcumin can alleviate kidney injury in ARDS rats, and its mechanism may be related to the increasing in SOD activity, reduction of oxidative stress, and inhibition of the activation of the NF-κB/NLRP3 inflammasome signaling pathway.
Male ; Rats ; Animals ; Rats, Sprague-Dawley ; NF-kappa B ; Actins ; Caspase 3 ; Curcumin ; Lipocalin-2 ; Toll-Like Receptor 4 ; Inflammasomes ; NLR Family, Pyrin Domain-Containing 3 Protein ; Reactive Oxygen Species ; Saline Solution ; Kidney ; Superoxide Dismutase