Background Choroidal neovascularization (CNV) is the primary pathogenic cause of many fundus diseases.Oxidative stress injury of retinal pigment epithelial (RPE) cells plays important role in angiogenesis of choroid new blood vessels.Oxidative stress injury can active p75NTR receptor, a member of tumor necrosis factors family,resulting in the proliferation of vascular endothelial cells.However, the mechanisms of vascular endothelial cell proliferation remain unclear.Objective This study was conducted to investigate the effect of p75NTR overexpression on CNV and the relative mechanism.Methods The ARPE-19 cell line was used in this study.RPE cells were transfected with p75NTR receptor overexpressed plasmid, and untransfected cells served as the control group.The transfected results were verified by reverse transcription-PCR and Western blot assay.Viability of the cells over time was determined in the p75NTR receptor plasmid transfected group by using BrdU assay.The percentage of apoptotic cells was detected by flow cytometry using Annexin V-FITC/PI fluorescence staining.The percentage of reactive oxygen species (ROS) expression in the cells was detected by using H2 DCFDA fluorescence and flow cytometry.Mitochondrial membrane potential and cytochrome C expression were examined under the confocal microscope.The protein expressions of cleaved caspase-3, Fas and VEGF were determined by Western blot assay.Results The relative expression level of p75 NTR receptor mRNA was (6.11 ±0.77) times higher than that of the control group, and relative expression level of p75NTR receptor protein in the cells in the p75NTR receptor plasmid transfected group was (7.42±0.48) times higher than that in the control group (t=11.49 and 23.17 ,both at P＜0.01).The absorbency values of the p75NTR receptor plasmid transfected group were (93.12±0.56) ％ , (86.30±0.66) ％ , (72.53-±0.86) ％ and (60.77 ±2.81) ％ in 12,24,36 and 48 hours after plasmid transfection, which were significantly lower than 100％ in the control group, and the apoptotic percentages were evidently higher than that in the control group (all at P＜0.05).The relative fluorescence intensity of ROS fluorescence in the p75NTR receptor plasmid transfected group was 2.4 times higher than that in the control group,showing significant difference (t=16.45, P＜0.01).The positive expressing rate of mitomarker (mitochondrial membrane potentials) was 100％ in the control group and (37.30± 2.06)％ in the p75NTR receptor plasmid transfected group, with significant difference between them (t =57.71,P＜0.01).The fluorescence intensity of cytochrome C expression was elevated in the p75NTR receptor plasmid transfected group compared with the control group.Compared with the control group,the expressing levels of cleaved caspase-3 ,Fas and VEGF165 proteins in the cells were significantly raised in the p75NTR receptor plasmid transfected group (all at P＜0.01).Conclusions Overexpression of p75NTR receptors in RPE cells leads to mitochondrial damage and cellular apoptosis and the secretion of VEGF protein, which sequentially promote CNV.P75NTR receptor may be another important regulation pathway in RPE oxygen damage.

Objective To provide a practicable method for the complete display and localization of the posterolateral structures (PLS) of the normal knee through MRI study. Methods 30 tibial bone specimens were observed to establish the bony landmark for localizing the knee. In 50 cadaver knees, the angles between lateral tibial plateau and the long axis of the individual structure of PLS were measured. Then the scan methods of the oblique MR images were determined based on above results. The routine and oblique scans of T 1WI were performed in 40 normal knees. The display effect and appearance of the PLS were observed on MRI. Results The lateral tibial plateau was a stable bony landmark for measuring and localizing of the knee. In the 40 normal knees, The fibular collateral ligament could be intactly displayed on 70? posterior coronal oblique images in 34 cases (85%). The popliteus could be better seen on either 45? medial sagittal oblique in 34 cases (85%) or 60? posterior coronal oblique planes in 36 cases (90%). The popliteofibular ligament could be intactly appreciated on both 60? posterior coronal oblique in 32 cases (80%) and 70? lateral sagittal oblique images in 34 cases (85%). Although the arcuate ligament and the fabellofibular ligament could occasionally be seen on routine and oblique images, but the display rate was lower. Conclusion The oblique MR imaging can intactly display the main structures of PLS, and can be useful in diagnosing the injuries in those structures.

In this study, we investigate the bioaccessibility of heavy metals (Cu, Pb, As, Cd and Hg) in wild Artemisia annua and use target hazard quotients (THQ) proposed by US Environmental Protection Agency to assess the health risk under the heavy metal exposure. The results showed that the bioaccessibility of Cu, Pb, As, Cd and Hg in A. annua are 0.77, 0.66, 0.46, 0.68 and 0, respectively, and that the value of THQ for adults and children were 0.030 and 0.025 calculated by risk assessment model. The results indicated that the heavy metals in A. annua were not able to be completely absorbed by human body and that their contents were in a safe range. In this study, by combining the bioavailability of heavy metal and health risk assessment, we assessed the security of heavy metals of wild A. annua, which will provide reference for the standard of heavy metals for medicinal materials.
Artemisia annua ; chemistry ; metabolism ; Consumer Product Safety ; Drug Contamination ; Humans ; Metals, Heavy ; analysis ; metabolism ; Risk Assessment ; Soil Pollutants ; analysis ; metabolism

Pulmonary fibrosis is a chronic and progressive lung disease that poses a threat to human health. Current treatment options are limited, highlighting the urgent need for more effective therapeutic strategies. Tetrandrine (TET), a bis-benzylisoquinoline alkaloid extracted from Stephania tetrandra, has been known for its anti-inflammatory and anti-fibrotic effects, but its specific mechanisms remain unclear. This study investigated the anti-fibrotic effects of TET in a chronic model of pulmonary fibrosis, aiming to delineate the molecular mechanisms underlying TET-mediated inhibition of fibroblast activation. The results showed that TET significantly alleviated the pathological changes in a murine model of multiple bleomycin-induced pulmonary fibrosis and effectively inhibited TGF-β1-induced fibroblast activation. Mechanistically, TET predominantly inhibited the TGF-β/SMAD signaling pathway and diminished intracellular reactive oxygen species (ROS) levels. Utilizing CRISPR-Cas9 library screening, we identified that angiotensin II type 1 receptor associated protein (AGTRAP) and membrane palmitoylated protein 6 (MPP6) played important roles in TET's suppressive impact of ROS levels, with the knockout of two genes attenuating TET's antifibrotic activity. All animal treatment procedures were approved according to the Committee on the Ethics of Animal Experiments of the Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences (IMB-20230406D507). This research not only elucidates the pharmacological mechanism of TET but also provides a novel therapeutic avenue for the treatment of pulmonary fibrosis.

Anesthetics, Intravenous ; administration & dosage ; pharmacology ; Animals ; Animals, Newborn ; Cerebral Cortex ; cytology ; metabolism ; Drug Synergism ; Female ; Fentanyl ; administration & dosage ; pharmacology ; Male ; Midazolam ; administration & dosage ; pharmacology ; Neurons ; metabolism ; Patch-Clamp Techniques ; Primary Cell Culture ; Rats ; Rats, Sprague-Dawley ; Voltage-Gated Sodium Channels ; drug effects

Objective To investigate the protein and mRNA expression of HO-1 in the lung tissue of asthmatic rat and the correlation between the percentage of blood carbon monoxide Hb(COHb)and the expressions of HO-1 in the lung tissue of asthmatic rat.Methods Twenty Sprague-Dawley(SD)male rats were randomly divided into 2 groups,control group and asthma group.Each group had 10 rats.The assessment of the percentage content of blood carbon monoxide Hb(COHb)wag performed.The total cell number and differentiation cell numbers in bronchoalveolar lavge fluid(BALF)and the inflammatory cells of airway wall were counted,the protein expressions of HO-1 in airway wall wag detected with immunohistochemistry technique.and the mRNA expressions of HO-1 in airway wall was detected with RTPCR.Results The expression of HO-1 was mainly located in airway epithelium and macrophage.The percentage of the cells in which protein of HO-1was expressed were(5.03±1.22)%,(27.14±4.68)%in two groups.The optical densities of mRNA expression of HO-1 were 0.323±0.05,0.68±0.02.The percent content of blood carbon monoxide Hb(COHb)were(0.45±0.35)%,(3.89±1.15)%.The protein and mRNA expressions of HO-1 and the percent content of COHb in asthmatic group were higher than those of the control group (P＜0.01 respectively).There was a significant positive correlation araong the protein expression of HO-1 in airway wall and the percent content of COHb(r=0.971,P＜0.001),mRNA expressions of HO-1 in lung tissue and the percent content of COI-Ib(r=0.897,P＜0.001).Conclusion The protein and mRNA expressions of HO-1 in the lung tissue,and the percent of COHb in blood were significantly mcreaged in asthmatic rat,which suggested HO-1 may play a significant role in the pathogenesis of asthma.

Acute Coronary Syndrome ; complications ; drug therapy ; Aged ; Hematoma ; complications ; Heparin, Low-Molecular-Weight ; adverse effects ; therapeutic use ; Humans ; Leg ; Male ; Thrombolytic Therapy ; adverse effects

Objective To investigate the expression of micro RNA-146a (miR-146a),TNF receptorassociated factor 6 (TRAF6) gene and IL-1 receptor-associated kinase 1 (IRAK1) gene in the peripheral mononuclear cells (PBMCs) of patients with ankylosing spondylitis (AS) and their relationship with the disease activity.The role of miR-146a,TRAF6,IRAK1 in the pathogenesis of AS was explored.Methods Expression of miR-146a,TRAF-6 and IRAK-1 in peripheral blood mononuclear cells was studied using realtime polymerase chain reaction (qRT-PCR) in 45 AS patients and 22 healthy controls.The indicators of disease activity adopted in this study were Bath ankylosing spondylitis disease activity index (BASDAI),erythrocyte sedimentation rate (ESR),C-reactive protein (CRP) level,and immunoglobulin (Ig).The relationship was analyzed in AS patients between the relative expression levels miR-146a,TRAF6,IRAK1 and BASDAI,ESR,CRP,Ig concentration.Non-parametric test,t test,One-way ANOVA,Pearson's and Spearman's correlation analysis were used for statistical analysis.Results ①The relative expression level of miR-146a which was observed in PBMCs of AS patients was significantly higher than that in normal control group [1.46(0.39,4.79)and 0.81(0.17,1.90),P＜0.05].The expression of miR-146a was significantly higher in active AS patients group than that in inactive patients [2.93(0.95,7.95) and 0.54(0.28,1.69),P＜0.05],there was no difference between the treatment group and without treatment group [1.28(0.31,2.37) and 2.22(0.49,7.71),P＞0.05].② There was significant difference in the relative expression level of IRAK-1 between AS patients and the normal control group.IRAK1 was significantly higher in AS patients than that in normal control group (1.4±0.7,1.1±0.4,P＜0.05).However,there was not difference between active AS patients group and inactive patients group as well as treated group and untreated group (1.5±0.9,1.4±0.5; 1.6±0.7,1.3±0.7,P＞0.05).③ TRAF6 expression was obviously lower in AS patients than that in normal control group (1.3±0.6,1.7±0.8,P＜0.05),and that was also significantly lower in the untreated group and active group than that in the normal control group (1.1±0.7,1.7±0.8; 1.1±0.5,1.7±0.8,P＜0.05).④ Signi-ficant positive correlation was observed between the miR-146a level and BASDAI,as well as duration of morning stiffness (r=0.557,P=0.000; r=0.363,P=0.018).The expression level of IRAK1 was significantly negative correlated with IgM (r=-0.313,P=0.046).Conclusion ① miR-146a expression is up-regulated in patients with AS,and it may be a potential useful marker for disease activity in AS patients; ② The abnormal expression of IRAK1,TRAF6 in AS patients may play a role in the pathogenesis of AS.

ObjectiveThe purpose of this study was to assess the efficacy,safety and optimal dose of tacrolimus monotherapy in patients with refractory lupus nephritis(LN) who were resistant to cyclophosphamide(CYC).MethodsA total of 14 LN patients (2 men and 12 women) with persistent proteinuria who were resistant to CYC treatment more than 8 g for half a year were enrolled.Tacrolimus was initiated at 2 mg/d (patient weight＜60 kg) or 3 mg/d(patient weight≥60 kg) which was administered in two divided doses.Prospective data on daily proteinuria,serum album level and serologic lupus activity were collected and followed for 6 months.ANOVA and Pearson correlation analysis were used for statistical analysis.Results Mean age at baseline was(30±9) years.Mean urinary protein decreased significantly from(6.2±5.1) g at baseline to (1.1±0.9) g at 6 months (F=16.21,P＜0.01).Mean serum album level increased significantly from (27.9±9.7) g/L at baseline to(37.8±2.2) g/L at 6 months(F=16.71,P＜0.01 ).Complete or partial response was observed in 86％ of patients receiving tacrolimus therapy.The effective dosage in this study was 0.03-0.06mg·kg-1·d-1 of the patients who had complete response or partial response to tacrolimus.The tacrolimus level in partially and completely responding patients was less than 3 ng/ml.There was no significant difference among blood tacrolimus levels of complete,partial,and no response patients [(1.6-±0.4),(2.0±0.6) and (22±1.1) ng/nl],respectively).No definite correlation was found between efficacy and tacrolimus level.Tacrolimus was well tolerated at current dose,besides one with new onset hypertension and one with alopecia.ConclusionOur results suggest that tacrolimus at low dosage and serum level is potentially effective and safe for the treatment　of patients with LN and persistent proteinuria resistant to CYC.The optimal dosage of tacrolimus for LN may　be 0.03-0.06 mg·kg-1·d-1.

OBJECTIVETo study the expression of heme oxygenase-1 (HO-1) gene and protein and the effect of budesonide (BUD) on the HO-1 expression in lung tissues in rats with asthma.

METHODSFifty male Sprague-Dawley rats were randomly divided into 5 groups: normal control, asthma model, dexamethasone (DXM)-, hemin (HO-1 challenger)-or BUD-treated asthma. The asthma model was prepared by ovalbumin sensitization and challenge. The rats were sacrificed 24 hrs after the last challenge. The blood COHb content,and the total cell count and the percentage of differential cells in bronchoalveolar lavage fluid (BALF) were measured. The expression of HO-1 protein and mRNA in lung tissues was detected with immunohistochemistry and RT-PCR, respectively. The airway inflammation situations were evaluated by histopathology.

RESULTSThe airway inflammatory cell infiltration in the DXM-, hemin- and BUD-treated asthma groups was remarkably alleviated compared with that in the asthma model group. Compared with the normal control group, the expression of HO-1 mRNA and protein in lung tissues and the blood COHb content in the asthma model and the DXM-, hemin- and BUD-treated asthma groups were significantly up-regulated. The DXM-, hemin- and BUD-treated asthma groups showed significantly increased expression of HO-1 protein and mRNA in lung tissues compared with the asthma model group. The blood COHb content in the DXM-and the hemin-treated asthma groups was significantly higher than that in the asthma model group.

CONCLUSIONSThe expression of HO-1 protein and mRNA in lung tissues and blood HO-1 activity increased in rats with asthma,suggesting that HO-1 may be involved in the pathogenesis of asthma. HO-1 may have a protective effect against the airway inflammation in asthmatic rats. BUD and DXM can up-regulate the expression of HO-1 protein and mRNA, thus providing protective effects against the airway inflammation in asthmatic rats.

Animals ; Asthma ; drug therapy ; enzymology ; Budesonide ; pharmacology ; therapeutic use ; Carboxyhemoglobin ; analysis ; Dexamethasone ; pharmacology ; Heme Oxygenase-1 ; analysis ; genetics ; Hemin ; pharmacology ; Lung ; enzymology ; Male ; RNA, Messenger ; analysis ; Random Allocation ; Rats ; Rats, Sprague-Dawley