OBJECTIVEThe aim of the present study is to explore the effects of exhaustive exercise-induced oxidative stress on the antioxidant capacity and diformability of rat red blood cells.

METHODSRats were divided into three group (n = 10): sedentary control (C), exhaustive running exercise (ERE) and moderate running exercise (MRE) groups. Animals in the ERE group started treadmill running at a speed of 20 m/min speed with a 5% gradient, and reached a speed of 25 m/min with gradient 15% in 20 min. Running was continued until exhaustion. MRE group rats running at a speed of 20 m/min with a 5% gradient for 40 min. The levels of free thiol in erythrocyte membrane protein, lipidperoxidation levels and membrane protein components were analyzed. The red blood cell deformability of different groups was also observed.

RESULTSThe results showed that red blood cells were damaged by severe oxidative stress and the anti-oxidative capacity decreased significantly under exhaustive exercise conditions. Besides, lipid peroxidation and protein sulfhydryl cross-link based clustering of membrane were found after exhaustive exercise, and polymers high molecular weight (HMW) was formed. The elongation index (EI) was found to decline significantly in the ERE group compared with the C and MRE groups under shear stress (control group, 0.41 +/- 0.01 at 3 Pa and 0.571 +/- 0.008 at 30 Pa; ERE group, 0.314 +/- 0.013 at 3 Pa and 0.534 +/- 0.009 at 30 Pa; P < 0.05 and P < 0.01, respectively).

CONCLUSIONThese exercise-induced oxidative injure result in a significant decrease in deformability of rat erythrocytes, which in turn leads to dysfunction in the microcirculatory.

Animals ; Disease Models, Animal ; Erythrocyte Deformability ; Fatigue ; metabolism ; physiopathology ; Male ; Oxidative Stress ; Physical Conditioning, Animal ; Rats ; Rats, Sprague-Dawley

Background Clinical studies indicated that the pathogenesis of most corneal dystrophy is associated with the mutation of the transforming growth factor beta-induced (TGFBI) gene.However,the molecular mechanism of mutated TGFBI gene in corneal dystrophy is unclear. Objective The present study was to investigate the expression of the TGFBI gene in human corneal tissue and cells in vitro.MethodsHuman corneal epithelial cells and keratocytes were cultured and passaged,and donor corneal tissue was obtained for the section preparation.RT-PCR was used to detect the expression of TGFBI mRNA in human corneal tissue and cells.Immunofluorescence was used to test the expression of the TGFBI protein in the human corneal tissue,and immunohistochemistry was used to test the expression of the TGFBI protein in human corneal epithelial cells and corneal stromal cells.ResultsRT-PCR analysis showed that TGFBI mRNA could be detected as a 1274 bp band in human corneal tissue and corneal stromal cells,but no TGFBI mRNA was observed in corneal epithelial cells.Immunofluorescence assay revealed that corneal stromal cells were positive ly expressed for the TGFBI protein,but the corneal epithelial cells did not express the TGFBI protein.Immunohistochemistry indicated that the expression of TGFBI was detected the red fluoressence in the cytoplasm of corneal stromal cells;however,no positive response was found in corneal epithelial cells.ConclusionsThe expression of the TGFBI gene occurs in human corneal stromal cells but not in the corneal epithelial cells.This result might be of helpful for studying the function and role of TGFBI gene in pathogenesis of corneal dystrophy.

Objective To explore the roles of frequency domain OCT in measuring the retinal thickness around the optic disc and optic disc parameters in early glaucoma diagnosis.Methods The optic disc parameters and retinal nerve fiber layer (RNFL) thickness in the 40 healthy volunteers (control group) and 85 cases of primary open angle glaucoma (POAG),including 36 patients as the early glaucoma subgroup and 49 patients as the glaucoma evolutum subgroup,were measured by frequency domain OCT.Then,the correlation analysis of RNFL thickness,optic disc parameters and the mean deviation (MD) of visual field in each group was performed,and the area under the curve was used to evaluate the diagnostic efficiency of RNFL thickness around the optic disc and optic disc parameters in the diagnosis of glaucoma.Results The RNFL thickness and the complete cycle mean RNFL thickness in the temporal,upper,nasal and inferior quadrant in the glaucoma patients were significantly lower than those in the controls (all P ＜ 0.05),and the above indexes in the glaucoma evolutum subgroup were significantly decreased compared with those in the early glaucoma subgroup (all P ＜ 0.05).There were statistically significant differences in the optic disc parameters between the groups except the optic disc area (all P ＜ 0.05).Pearson correlation analysis showed the RNFL thickness and the complete cycle mean RNFL thickness in the temporal,upper and inferior quadrant were negatively correlated with the MD in the glaucoma patients (all P ＜ 0.05),and the parameter of optic cup volume and cup/disc area ratio were positively correlated with the MD (both P ＜ 0.05),and the rim area,rim volume and disc volume were negatively correlated with MD (all P ＜ 0.05).The ROC curve analysis showed that the largest area under the curve of RNFL thickness in the inferior quadrant of the optic disc region was 0.886,and the specificity and sensitivity was 0.775 and 0.924,respectively.Moreover,the area under the curve of the optic cup/optic disc area was the largest,with sensitivity and specificity of 0.741 and 0.815,respectively.Conelusion OCT for measuring optic disc structure and RNFL thickness can be used for early diagnosis of glaucoma,and it has a high sensitivity and specificity.

OBJECTIVETo study the inhibitory effect of paeoniflorin (PAE) on TNF-α-induced TNF receptor type I (TNFR1)-mediated signaling pathway in mouse renal arterial endothelial cells (AECs) and to explore its underlying molecular mechanisms.

METHODSMouse AECs were cultured in vitro and then they were treated by different concentrations PAE or TNF-α for various time periods. Expression levels of intercellular cell adhesion molecule-1 (ICAM-1) were detected in the normal group (cultured by serum-free culture media), the TNF-α group (cultured by 2-h serum-free culture media plus 6-h TNF-α 30 ng/mL), the low dose PAE group (cultured by 2-h PAE 0.8 μmo/L plus 6-h TNF-α 30 ng/mL), the middle dose PAE group (cultured by 2-h PAE 8 μmol/L plus 6-h TNF-α 30 ng/mL), the high dose PAE group (cultured by 2-h PAE 80 μmol/L plus 6-h TNF-α 30 ng/mL) with Western blot analysis. Nuclear translocation of transcription factor NF-κB (NE-κB) was detected in the normal group (cultured by serum-free culture media), the TNF-α group (cultured by 2-h serum-free culture media plus 45-mm TNF-α 30 ng/mL), and the high dose PAE group (cultured by 2-h PAE 80 μmol/L plus 45-min TNF-α 30 ng/mL) by immunofluorescent staining. Expression levels of the phosphorylation of extracellular signal-regulated (protein) kinase (ph-ERK) and p38 (ph- p38) were detected in the normal group (cultured by serum-free culture media) and the high dose PAE group (2-h PAE 80 μmol/L culture) by Western blot. NF-κB inhibitor-α (IκBα) protein expressions were detected in the normal group (cultured by serum-free culture media), the TNF-α group (cultured by 2-h serum-free culture media plus 30-min TNF-α 30 ng/mL), the high dose PAE group (cultured by 2-h PAE 80 μmol/L plus 30-min TNF-α 30 ng/mL), the p38 inhibitor group (SB group, pretreatment with SB238025 25 μmol/L for 30 min, then treated by PAE 80 μmol/L for 2 h, and finally treated by TNF-α 30 ng/mL for 30 min), the ERK inhibitor group (PD group, treated by PD98059 50 μmol/L for 30 min, then treated by PAE 80 μmol/L for 2 h, and finally treated by TNF-α 30 ng/mL for 30 min) by Western blot.

RESULTSCompared with the normal group, ICAM-1 protein expression levels obviously increased (P < 0.01). Compared with the TNFα group, ICAM-1 protein expression levels were obviously inhibited in the high dose PAE group (P < 0.05). Protein expression levels of ph-p38 and ph-ERK were obviously higher in the hIgh dose PAE group (P < 0.05). Compared with the normal group, IκBα protein expression levels obviously decreased in the TNF-α group (P < 0.01). Compared with the TNFα group, TNF-α-induced IκBα degradation could be significantly inhibited in the high dose PAE group (P < 0.01); the inhibition of PAE on IκBα degradation could be significantly inhibited in the SB group (P < 0.05). NF-κB/p65 signal was mainly located in cytoplasm in the normal group. NF-κB/p65 was translocated from cytoplasm to nucleus after stimulated by 45 min TNF-α in the TNF-α group, while it could be significantly inhibited in the high dose PAE group.

CONCLUSIONSPAE inhibited TNF-α-induced expression of lCAM-1. Its action might be associated with inhibiting TNFR1/NF-κB signaling pathway. p38 participated and mediated these actions.

Animals ; Cells, Cultured ; Endothelial Cells ; cytology ; drug effects ; Glucosides ; pharmacology ; Intercellular Adhesion Molecule-1 ; metabolism ; Mice ; Monoterpenes ; pharmacology ; NF-kappa B ; metabolism ; Receptors, Tumor Necrosis Factor ; metabolism ; Signal Transduction ; drug effects ; Tumor Necrosis Factor-alpha ; pharmacology

The aim of this study is to investigate the critical factor affecting the properties of PLGA microspheres fabricated by a solid-in-oil-in-water (S/O/W) emulsion technique with BSA as a model protein. Prior to encapsulation, the BSA microparticles were fabricated by a modified freezing-induced phase separation method. The microparticles were subsequently encapsulated into PLGA microspheres by S/O/W emulsion method, then Motic BA200 biological microscope, confocal laser scanning microscope, scanning electron microscope were used to observe the structure of S/O/W emulsion and PLGA microspheres. The protein content extracted or released from BSA microspheres was measured by Bradford protein assay method. It was found that NaCl added in the outer aqueous phase effectively suppressed material exchange between the inner and outer phase of S/O/W emulsion. Then, the structure and permeability of obtained microspheres were influenced. As a result, with the increase of NaCl concentration in the outer aqueous phase, the encapsulation efficiency of microspheres significantly increased from 60% to more than 85%, the burst release of microspheres reduced from 70% to 20%, and the particle size decreased from 103 microm to 62 microm. Furthermore, the rehydration of encapsulated protein was also retarded and then integrity of BSA was successfully protected during encapsulation process. In vitro release test showed that BSA released from PLGA microspheres in a sustained manner for more than 30 days.
Delayed-Action Preparations ; Drug Compounding ; Emulsions ; chemistry ; Lactic Acid ; administration & dosage ; chemistry ; Microscopy, Confocal ; Microscopy, Electron, Scanning ; Microspheres ; Oils ; Particle Size ; Polyglycolic Acid ; administration & dosage ; chemistry ; Serum Albumin, Bovine ; administration & dosage ; chemistry ; Sodium Chloride ; chemistry ; Water

OBJECTIVETo explore the anatomical structures, and depth and direction of needling at Jingming (BL 1), so as to provide anatomical basis for its clinical application.

METHODSForty-eight adult orbital specimens were observed by dissection.

RESULTSWhen a needle was vertically inserted into Jingming (BL 1), the needle tip will past through the skin, subcutaneous tissue, medial palpebral ligament, medialis rectus and orbital adipose body. Above the body of the needle, there are ophthalmic artery, anterior ethmoidal artery and nasociliary nerve. The average distance between the skin at the punctured point and the anterior ethmoidal artery is (18.25 +/- 4.45) mm, with an angle of (12.5 +/- 5.5) degrees, and the average distance between the skin at the punctured point and the optic nerve tunnel frontal point is (43.37 +/- 7.84) mm.

CONCLUSIONTo avoid bleeding caused by injuring the anterior ethmoidal artery, acupuncture at Jingming (BL 1) should avoid deeply inserting needled back-upwards and upwards, and the needling depth should not exceed 30.36 mm to avoid injury of the optic nerve tunnel frontal point.

Acupuncture Points ; Female ; Humans ; Male ; Orbit ; anatomy & histology

Klebsiella pneumoniae was cultured followed by the preparation and immunoactivity elucidating of its polysaccharide (CPS). The lysis of cell is the first key step in the preparation, under the co-action of trypsin, lysozyme and NP-40, the cell lysed within 2h, then the lysate was concentrated by ultrafiltration which serves as concentrating and partial purifying action simultaneously. Crude CPS was got by ethanol precipitation, then purified through the Ion-exchange and gel filtration, the purity of CPS was judged by the gel filtration and agarose gel electrophoresis. The effect of CPS on the cell immunoactivity was studied in detail, the results show that CPS possesses bidirectional immunoregulation on the spleen cells of mice, that is, low concentration of CPS can stimulate the immune response while the high concentration manifests the inhibition significantly. The investigation results will benefit on the exploitation of the CPS.
Animals ; Bacterial Capsules ; chemistry ; Culture Media ; Klebsiella pneumoniae ; chemistry ; immunology ; Lymphocyte Activation ; drug effects ; immunology ; Lymphocytes ; immunology ; Mice ; Mice, Inbred BALB C ; Polysaccharides, Bacterial ; immunology ; isolation & purification ; Spleen ; cytology

Objective To evaluate the clinical effect of embolization of cerebral dural atreriovenous fistulas (cDAVF) of the eavemous sinus through the superior ophthalmic vein approach. Methods Twnety-seven patients with eDAVF of the cavernous sinus were embolized through the superior ophthalmic vein approach. Cerebral angiography and follow-up examination of the patients were performed to evaluate the effect ofernbolization. Results The fistulae showed complete angiographic disappearance in 15 patients, and 12 patients had blood velocity flow reduction at the fistula orifice. Ocular proptosis and chemosis deteriorated transiently in 11 patients after the procedure. The patients were followed-op for 3 to 48 months, and clinical cure was achieved in 17 patients, and 10 showed significant symptom relief. Conclusion cDAVF of the cavernous sinus can be effectively embolized through the superior ophthalmic vein approach.

Objective To evaluate the association between resting heart rate (HR) and all-cause death and coronary heart disease (CHD) events in the Chinese cohort. Methods Data were obtained from the PRC-USA Cooperative Study on Cardiovascular and Cardiopulmonary Epidemiology. Baseline screen surveys were conducted in 1983 and 1984 from people aged 35 to 59 years living in urban or rural areas of Beijing and Guangzhou. Follow-up visits were performed for end point events of all-cause death and first CHD events every two years till 2005. Resting HR was determined from 5 consecutive intervals between R waves on the 12-lead electrocardiogram. Results A total of 9856 (4805 males) people were included in the study and the mean follow up duration was 16. 2 years. There were 1523 deaths, including 200 CHD events during the follow up period. Mean resting HR was 67. 9 beat per minute (bpm) in men and 71. 6 bpm in women respectively which had a trend to increase with aging. Cox Proportional Hazards model indicated the relative risk of all-cause death increased constantly with the increase of HR percentile after control of age, fasting glucose, serum cholesterol, serum triglyceride, body mass index, systolic blood pressure and diastolic blood pressure. With HR 60-89 bpm as control group, the relative risk and 95% confidence interval in group HR <50 bpm,50-59 bpm,90-99 bpm and ≥100 bpm were 0. 76(0. 49-1. 17) ,0. 87 (0. 75-1. 02) , 1. 33 ( 1. 06-1. 68) ,1.48 ( 1. 03-2. 14) respectively. However there was no significant correlation between HR and CHD events in studied population. Conclusion The risk of total death increased significantly in people with HR≥90 bpm suggesting higher resting heart rate might be an independent risk factor for all-cause death in the Chinese population.

Objective To explore the changes of hemodynamics in pregnant women with uncomplicated dichorionic(DC)twin pregnancy and monochorionic(MC)twin pregnancy,so as to better perform prenatal monitoring.Methods A study was conducted to collect 64 pregnant women with uncomplicated twin pregnancy(41 cases of DC and 23 cases of MC)in the Obstetrics and Gynecology Hospital,Fudan University from May 2020 to Sept 2021,and 144 pregnant women with uncomplicated singleton pregnancies in the same period were selected as the control group.During the second trimester(20-28 weeks),conventional echocardiography was performed in pregnant women,and singleton pregnancy was used as the control.The left ventricular systolic,left ventricular diastolic,hemodynamic and cardiac structural parameters of DC and MC twin pregnancy were studied.Results At 20-28 weeks of gestation,compared with singleton pregnancies,the heart rate,mean arterial pressure,cardiac output,cardiac index,stroke volume,left ventricular mass,stroke work index parameters of twin pregnancy were significantly increased,and the total vascular resistance was significantly decreased,and the differences were statistically significant(P<0.05).The distribution of maternal hemodynamic parameters was similar in pregnant women with DC twin and MC twin pregnancy.Compared with DC twin pregnancy,MC twin pregnancy showed a significant increase in cardiac output(5.76 L/min vs.5.36 L/min,P=0.031).Total vascular resistance significantly decreased(1 270 vs.1 407,P=0.037).Conclusion Compared with singleton pregnancy,the hemodynamics of twin pregnancy significantly changed,which ensured the growth and development of the fetus by providing sufficient uteroplacental circulation.The cardiovascular adaptation patterns of DC and MC twin pregnancy were similar,but MC twin pregnancy had higher cardiac output and lower total vascular resistance.Monitoring the cardiac function of twin pregnancy,especially MC twin pregnancy,is very important for the safety of pregnant women.