AIM:To investigate the effects of MG132,one of the proteasomal peptide aldehydes inhibitors,on lipopolysaccharide(LPS)-induced nuclear transcription factor-kappa B(NF-?B) activation,the production of nitric oxide(NO) and tumor necrosis factor-alpha(TNF-?),as well as the expression of inducible nitric oxide synthase(iNOS) in murine macrophage line RAW264.7.METHODS:Reporter gene assay was used to examine the activity of NF-?B by pNiFty-SEAP/HEK293 cells,which were transfected with the pNiFty reporter plasmid into human embryo kidney cells(HEK293).Fluorescence substrate DAF-2DA was used to testify NO level in Raw264.7 cell line induced by LPS.Furthermore,the secretion of TNF-? was examined by ELISA.Western blotting was used to reveal the expression of iNOS and I?B-?.RESULTS:MG132 significantly decreased the secretion of TNF-? induced by LPS,with the inhibitory rates of 36.7% and 60.4% to 5 ?mol/L and 10 ?mol/L MG132,respectively.The pro-inflammatory mediator NO production was decreased in a dose-dependent manner with the inhibitory rates increasing from 29.5%(2 ?mol/L) to 55.9%(10 ?mol/L).Pretreatment with MG132 reduced the expression of iNOS,but restored the I?B restrain caused by LPS treatment.Observed by a reporter gene assay,TNF-?-induced NF-?B activity was decreased gradually by addition of increasing concentration of MG132(2.5-10 ?mol/L).CONCLUSION:Our results suggest an anti-inflammation effect of MG132 by the suppression of LPS-induced the production of pro-inflammatory mediators including NO and TNF-?,and the expression of iNOS,which probably mediates the blockage of I?B degradation and NF-?B activation.

Amifostine ; pharmacology ; Anemia, Aplastic ; chemically induced ; pathology ; prevention & control ; Animals ; Benzene ; toxicity ; Dose-Response Relationship, Drug ; Male ; Mice

0.05). Serum IL-10 and IL-10/TNF? ratio in the dead were significantly higher than that in the survivors( P 0.05). Conclusions The septic patients with high serum IL-10 or IL-10/TNF? ratio fared with poor prognosis.

Objective To explore the clinical characteristics ,diagnosis, treatment and prognosis of phaeohyphomycosis. Methods Clinical data were collected, including history, physical examination, cranial and spinal imaging. Brain biopsy was performed. Data of the pathology and incubation of brain tissue were analyzed. Responsiveness to treatment was followed up. Results A previously healthy three and half years old boy was presented to our unit, with a three- month history of recurrent headache, vomiting, progressive paraplegia accompanied by urinary continence and constipation. A computed tomogram scan and magnetic resonance imaging of the brain revealed multiple lesions located in the region of the parietal - occipital lobes, periventricular area and frontal lobe, with prominent surrounding edema and irregular peripheral enhancement of the mass after the administration of contrast materials. A cerebral biopsy was performed and the pathological report was cerebral phaeohyphomycosis. The culture of the tissue and cerebrospinal fluid grew a same fungus identified as exo-phiala dermatitidis. The patient's response to therapy was poor, the parents of the boy gave up therapy, and the boy died 1 month later. Conclusions Cerebral Phaeohyphomycosis caused by Exophiala dermatitidis is rare, but the most serious form of fungus infection. Pathology and incubation of the tissue are essential for diagnosis. There is no curative therapy and the prognosis is poor.

The anti-aging effect of Yang Shou Dan (YSD) was studied. It was shown that YSD could obviously increase the activity of superoxicde dismutase (SOD) in erythrocyte, decrease lipid peroxides (LPO) in serum and in liver tissue and reduce the content of lipofucin in myocardium of mice. It could also prolong the mean life span, the half lethal period and the longest life time of drosophila melanogater. It is suggested that YSD exerts an anti-aging effect to a certain extent.

Background Leber hereditary optic neuropathy (LHON) is mitochondrial DNA (mtDNA) disease and mainly leads to optical nerve degeneration.Its primary mechanism is synthesis disorder of DN4 protein due to variation of mtDNA 11778 locus.So to construct a vector with exogenous normal ND4 and transfect into mitochondria is a key of gene therapy for LHON.Objective This study was to investigate the in vitro transfection of adeno-associated virus (AAV)-ND4 gene into mitochondria.Methods Human renal epithelial cell lines transfected adenovirus E1A (293 cells) were regularly cultured and divided into two groups.Framework plasmids of recombinant AAV-ND4 or simple AAV2 were added to the cell medium respectively.The expression of ND4 in cells were located 12,24,36 and 48 hours after transfected by Y03 dual fluorescent quantum dots staining.The positive response for ND4 showed the green fluorescence.Results Cultured 293 cells grew well with 80％ confluence.Abundant green fluorescence particles were seen in cytoplasm in the AAV-ND4 transfected group,but only red fluorescence from mitochondrial protein was seen in the simple AAV transfected group under the fluorescence microscope.Conclusions Exogenous ND4 protein can been successfully transfected into mitochondria using the ND4 gene constructed AAV.This result provides experimental evidence for the further study on gene therapy of LHON.

Adult ; Biopsy, Needle ; Diagnosis, Differential ; Humans ; Kidney ; pathology ; Kidney Diseases ; pathology ; therapy ; Male ; Nephritis, Interstitial ; pathology ; Renal Dialysis ; Sarcoidosis ; pathology ; therapy ; Tuberculosis, Renal ; pathology

OBJECTIVETo explore the molecular mechanism of pain associated with chronic prostatitis and chronic pelvic pain syndrome (CP/CPPS) in the rat model of prostatic inflammation.

METHODSThirty-six male SD rats were equally randomized to an experimental and a control group, the former injected with 50 μl of 3% λ-carrageenan into the ventral prostate to make the model of non-bacterial prostatic inflammation, while the latter with the same volume of sterile saline solution. At 1, 2 and 4 weeks after modeling, the prostate, L6-S1 dorsal root ganglion (DRG) and spinal cord were harvested for examination of the expressions of the nerve growth factor (NGF), transient receptor potential ankyrin 1 (TRPA1), and calcitonin-gene-related peptide (CGRP) by immunohistochemistry and Western blot.

RESULTSThe expressions of NGF, TRPA1 and CGRP in the prostatic tissue were all significantly increased in the experimental group as compared with the control (P <0.05), with a gradual decrease with the prolonging of time (P <0.05). In the L6-S1 DRG and spinal cord, the expressions of NGF, TRPA1 and CGRP exhibited no significant differences between the experimental and control groups at 1 week after modeling (P >0.05) and kept at high levels in the experimental group at 2 and 4 weeks, though not significantly different from those at 1 week (P >0.05). Statistically significant differences were observed in the expressions of the three proteins in the experimental rats among different time points (P <0.05), but not between the two groups at any time point (P >0.05).

CONCLUSIONThe molecular mechanism of CP/CPPS can be evaluated in the rat model of prostatic inflammation established by injecting λ-carrageenan into the prostate. TRPA1 may play an important role in connecting the upstream and down-stream pathways of CP/CPPS-associated pain.

Animals ; Calcitonin Gene-Related Peptide ; metabolism ; Carrageenan ; Chronic Disease ; Chronic Pain ; metabolism ; Ganglia, Spinal ; metabolism ; Humans ; Male ; Nerve Growth Factor ; metabolism ; Pelvic Pain ; metabolism ; Prostatitis ; chemically induced ; metabolism ; Rats ; Rats, Sprague-Dawley ; Spinal Cord ; metabolism ; TRPA1 Cation Channel ; TRPC Cation Channels ; metabolism

OBJECTIVETo study the effects of erythropoietin (EPO) on cardiomyocyte apoptosis and endoplasmic reticulum stress (ERS)-related proteins, glucose-regulated protein 78 (GRP78) and C/EBP homologous protein (CHOP), in neonatal rats with asphyxia.

METHODSA total of 120 newborn Sprague-Dawley rats (7 days old) were randomly divided into sham-operated (n=40), asphyxia (n=40) and EPO-treated asphyxia groups (n=40). A neonatal rat model of normobaric asphyxia was established in the asphyxia and EPO-treated asphyxia groups. The rats in the EPO-treated asphyxia group received intraperitoneal injection of recombinant human erythropoietin (500 U/mL) immediately after the model was established, while the other two groups received the same volume of normal saline (0.9%). Heart blood and myocardial tissue samples were collected from 8 rats in each group at 2, 6, 12, 24 or 48 hours after the model was established. Serum creatine kinase (CK) and lactate dehydrogenase (LDH) levels were measured; cardiomyocyte apoptosis was evaluated, and expression of myocardial GRP78 and CHOP was measured.

RESULTSCompared with the sham-operated and EPO-treated asphyxia groups, the asphyxia group had significantly increased serum CK and LDH levels, number of apoptotic cells, and expression of myocardial GRP78 and CHOP at each time point (P<0.01), and all the indices were significantly higher in the EPO-treated asphyxia group than in the sham-operated group (P<0.01). At 24 hours after asphyxia, the expression of myocardial CHOP was positively correlated with the myocardial apoptosis index (r=0.944, P<0.01).

CONCLUSIONSEPO exerts a protective effect on the myocardium of neonatal rats with hypoxic-ischemic injury by regulating ERS-related proteins GRP78 and CHOP and reducing cardiomyocyte apoptosis.

Animals ; Animals, Newborn ; Apoptosis ; drug effects ; Asphyxia Neonatorum ; drug therapy ; pathology ; Creatine Kinase ; blood ; Endoplasmic Reticulum Stress ; physiology ; Erythropoietin ; pharmacology ; Heat-Shock Proteins ; analysis ; L-Lactate Dehydrogenase ; blood ; Myocytes, Cardiac ; drug effects ; pathology ; Rats ; Rats, Sprague-Dawley ; Transcription Factor CHOP ; analysis

Objective:To analyze the echocardiographic features of operation-related aorto-cardiac fistula(ACF) after surgery or transcatheter procedure, to explore the value of echocardiography on diagnosis of operation-related ACF, and summarize the key points of its diagnosis.Methods:Eight patients with operation-related ACF who were admitted to Fuwai Hospital were collected from July 2002 to December 2020. Echocardiographic features of the 8 patients with operation-related ACF were reviewed and analyzed. The diagnosis methodology was summarized.Results:Of the 8 patients with operation-related ACF, 3 had aortic right atrial fistula and 5 had aortic right ventricular fistula. The fistula was single, which can be located but not limited to the aortic sinus. The median size of the fistula was 4 mm (range: 3-5 mm). There was no aneurysmal dilation of the aortic sinus where the fistula was located. The fractured end of the fistula did not thin, and the fistula had a regular shape. Six of the 8 patients undertook cardiac catheterization and occlusion. One patient received surgical repairment. One patient was treated conservatively.Conclusions:Echocardiography can help diagnose operation-related ACF and provide valuable information for further clinical diagnosis and treatment. Sonographers should avoid missed diagnosis and pay attention to distinguishing from other causes of aortic-cardiac shunt disease.