Oxidative stress, autophagy, and apoptosis induced by doxycycline in loach fin cells in vitro.

Cytotoxicity, molecular function disorder, mitophagy, and apoptosis were studied in loach fin cells in vitro after exposure to doxycycline (DOX). The semi-lethal concentration of DOX in loach cells was calculated as 668.96 ± 2.83 mol/L. Loss of cell viability and increases in vacuoles and autolysosomes were evident in cells exposed to DOX at 200 and 400 µmol/L, and apoptotic bodies occurred at 600 µmol/L. In addition, Superoxide Dismutase (SOD), catalase (CAT), Na+-K+-ATPase, and Ca2+-ATPase activities increased significantly in cells exposed to 200 µmol/L DOX, and dose-dependent inhibitory effects on activities were observed in cells exposed to 400 and 600 µmol/L DOX. Quantitative gene expression showed that 400 and 600 µmol/L DOX could induce caspase-3- and caspase-8-mediated apoptosis as well as caspase-activated DNase in loach cells. Transcriptome sequencing in DOX vs. control groups found 16,288 differentially expressed genes, among which protein binding (2633, 31.91%) was the most significant in Gene Ontology terms. Furthermore, 11,930 genes were enriched in 298 Kyoto Encyclopedia of Genes and Genomes (KEGG)pathways. The top three upregulated pathways included "lysosome", "protein processing in endoplasmic reticulum", and "proteasome". FPKM analysis indicated that most genes associated with autophagy and in "protein processing in the endoplasmic reticulum", "TNF signaling pathway", and "NF-kappa B signaling pathway" were upregulated. This suggests that at lower concentrations, DOX induces reactive oxidative species (ROS) in loach fin cells to reduce cell proliferation. ROS in turn stimulate oxidant stress, ion excretion capability and mitophagy to maintain cell homeostasis. Apoptosis was induced in cells subjected to higher concentrations of DOX. The transcriptome data and pathways determined in this study will provide a foundation for the analysis of DOX toxicity in loach cells, which must be examined thoroughly to further understand the cytotoxic mechanism of antibiotics in fish cells.

Determination of seven components in Pifubingxuedu tablet by HPLC method / 药学实践杂志

Objective To establish a HPLC method for simultaneous determination of seven constituents in Pifubingxuedu tablet.Methods The determination was performed on Kromasil-C18 column (4.6 mm × 250 mm, 5 μm).The mobile phase was acetonitrile-0.5％ phosphoric acid aqueous solution with a flow rate of 0.8ml/min.The detector was PDA and detection wavelength was set at 245,325,403 nm, respectively.Results A method has been established for the determination of chlorogenic acid, hydroxy Safflower Pigment A, paeoniflorin, forsythiaside A, ferulic acid, berberine and glycyrrhizin acid in one run by HPLC.Their average contents and RSD in each Pifubingxuedu tablet were 0.299 5 mg/tablet (2.25％);0.700 0 mg/tablet(1.33％);0.429 2 mg/tablet (1.21％);0.039 1 mg/tablet (2.34％);0.014 8 mg/tablet(2.23％);0.209 0 mg/tablet (2.06％);0.272 7 mg/tablet (2.68％), respectively.Conclusion The established method is simple, convenient, accurate and reliable.It is suitable for the quality control of Pifubingxuedu tablet.

Diagnosis and treatment for 2 cases of tuberous sclerosis complex with pulmonary lymphangioleiomyomatosis / 中南大学学报(医学版)

To improve the diagnosis and treatment for tuberous sclerosis complex (TSC) with pulmonary lymphangioleiomyomatosis, a retrospective analysis was performed based on the clinical data of 2 patients with such disease. Both of them have typical thin-walled cystic lesion throughout the lung field, renal angioleiomyolipoma, and various degrees of skin lesions. Central nervous system is involved in one patient. Lesions in the lung and kidney in one patient were improved significantly after 5 months of rapamycin treatment. The clinical phenotypes were diverse in TSC patients. The CT imaging showed typical characteristics when the lung was invaded by the tumor. When a patient was diagnosed as pulmonary lymphangioleiomyomatosis, we should pay attention to the clinical screening of TSC. Rapamycin is an effective and safe treatment for this disease.

Tauroursodeoxycholic acid suppresses endoplasmic reticulum stress in pulmonary tissues of intermittent hypoxia mice / 中南大学学报(医学版)

OBJECTIVE@#To explore the mechanism of tauroursodeoxycholic acid (TUDCA) in suppressing apoptosis in pulmonary tissues of intermittent hypoxia (IH) mice model.
@*METHODS@#A total of 32 C57 mice were randomly divided into a control group, a TUDCA group, an IH group and an IH+TUDCA group (8 mice per group). The mice were put in specially designed chambers and exposed to IH treatment for 4 weeks. In the chambers, oxygen levels repeatedly decreased from 21% to 10% and recovered from 10% to 21%, lasting for 8 hours in every day. After 4 weeks of IH exposure, the expression levels of caspase-12 and cleaved caspase-3 in pulmonary tissues were detected by Western blot. Meanwhile, the expression levels of glucose regulated protein-78 (GRP78) and CCAAT/enhancer-binding protein homologous protein (CHOP) were quantified by Western blot, immunochemistry and real-time PCR.
@*RESULTS@#Compared with the control group, the expression levels of caspase-12, cleaved caspase-3, GRP78 and CHOP were increased in the IH group (all P<0.01). TUDCA treatment could reduce these proteins expression (all P<0.05).
@*CONCLUSION@#Endoplasmic reticulum stress-mediated apoptosis can be activated in pulmonary tissues after chronic IH exposure, and TUDCA can reduce the cellular apoptosis via suppressing endoplasmic reticulum stress.

Diagnostic value of endobronchial ultrasound-guided transbronchial needle aspiration in elderly patients with bronchogenic carcinoma / 中华老年医学杂志

Objective To evaluate the diagnostic value and safety of endobronchial ultrasoundguided transbronchial needle aspiration (EBUS-TBNA) in elderly patients with bronchogenic carcinoma.Methods From August,2013 and July,2014,31 patients aged 65 years and over with space-occupying mass and enlarged mediastinal lymph nodes detected by CT or positron emission tomography-computed tomography (PET-CT) underwent EBUS-TBNA.Rapid onsite cytology evaluation was not performed.Results There were 26 males and 5 females in this study,aged 65-77 years (70.1 years old on average).In 31 patients,70 samples were obtained from lymph nodes (LNs) and 4 samples were obtained from intrapulmonary lesions.29 cases were diagnosed as lung cancer,and 2 cases had false-negative diagnoses.The sensitivity and specificity rates of EBUS-guided TBNA method were 93.6％ and 100.0％,respectively.No major complications were observed in this series.Conclusions EBUS-TBNA is an safe and effective method in diagnosing bronchogenic carcinoma in elderly patients.

Two cases of pulmonary thromboembolism associated with protein C and protein S deifciency and literature review / 中南大学学报(医学版)

To explore the clinical manifestations, diagnosis and treatment of pulmonary thromboembolism associated with protein C (PC)/protein S (PS) deifciency. Two male patients 29 and 26 years old diagnosed with PC deifciency and/or PS deifciency were retrospectively analyzed and related literatures were reviewed. The most common symptoms were pain in the lower limbs with chest pain or decreased vision. Color dopper lfow imaging (CDFI) showed lower deep venous phlebothrombosis. Multislice CT angiography (CTA) revealed pulmonary embolism. The level of serum homocysteine (HCY) increased and the level of plasma PC/PS content decreased to PC 57.4%, and PS 28.9%in patient 1, while PS 33.4%in patient 2. Poor routine anticoagulant response was observed. After the diagnosis of PC/PS deficiency, vitiamin B6 and B12 anticoagulant therapy was added, and the symptoms in the patients improved significantly. Congenital thrombophilia should be taken into consideration for young patients with lower deep venous thrombosis and pulmonary embolism which occur recurrently without obvious predisposing causes before 40. Plasma PC/PS concentrations or activity help a lot in the diagnosis and treatment.

Clinical features of primary Sj(o)gren syndrome associated with interstitial lung disease / 中国医师进修杂志

Objective To investigate the clinical features of primary Sj(o)gren syndrome (pSS) with interstitial lung disease (pSS-ILD) so as to raise the clinical diagnosis Level. Methods The clinical data were collected from 58 patients with pSS,who were admitted from March 2006 to March 2009. The patients were divided into ILD group (27 cases) and non-ILD group (31 cases). Rheumatoid factor(RF),C-reactive protein (CRP), protein electrophoresis and complement C3, C4 in serum were measured by immunoturbidimetric methods. Antinuclear antibodies (ANA) and anti-ENA antibodies were measured by indirect immunofluorescence technique and western blot. CA125 was detected by enzyme-linked immunosorbent assay and erythrocyte sedimentation rate (ESR) was detected by WS method. Pulmonaly function tests and radiology examination were performed. Results Compared with those in non-ILD group,the percentages of dry mouth,dry eye, rampant caries and velcro crackles were significantly higher in ILD group,anti-SSA antibody, ESR,CRP, CA125 and title of γ-gloulin was significantly higher in ILD group. In ILD group,diffusion dysfunction was 18 cases (66.7%),restricted ventilation dysfunction was 14 cases (51.9%),blocked ventilation dysfunction was 6 cases (22.2%), incorporated ventilation dysfunction was 8 cases(29.6%), forced vital capacity, forced expiratory volume in one second, maximum midexpiratory flow, DLCO were significantly lower in ILD group than those in non-ILD group (P ＜ 0.01). Moreover,high resolution computerized tomography (HRCT) was more sensitive than chest X-ray in the diagnosis of pSS-ILD.Conclusions The presence of pSS-ILD highly associates with the activity of pSS. Pulmonary diffusion ventilation function and HRCT play an important role in the diagnosis of pSS-ILD.

Administration of sirolimus affects vein graft neointima hyperplasia / 北京大学学报(医学版)

Objective: To investigate the effect of Sirolimus on vein graft neointima hyperplasia via oral administration compared with local delivery, and find out an effective and safe way to provide support for clinical application. Methods: A rabbit external jugular vein-to-common carotid artery model was established. Twenty-four healthy rabbits were divided into 4 groups at random: blank-control group, F-127 control group, group 3 that received locally applied slow-releasing Sirolimus with F-127, group 4 that received oral Sirolimus (the commercial name Rapamune). The ratio of intima to medium thickness and re-stenosis rate (ratio of lumina to lumina plus intima area) were measured, PCNA positive cells by immunohistochemical staining were detected to indicate the degree of cell proliferation, and apoptosis cells detected by TUNEL. Results: Compared with blank-control group, neointima hyperplasia was inhibited significantly in group 3 and group 4 intima thickness were (90.11?10.99)?m versus (29.38?10.45) ?m, (18.29?9.03)?m, respectively. Re-stenosis rate was reduced (lumina area/ total area ratio were 0. 58?0.11 versus 0.80?0.16, 0.77?0.16, respectively). Proliferation of VSMC was inhibited (cell proliferation indexes were 31.03%?6.80% versus 20.32%?9.19%, 16.22%?5.85%, respectively) and cell apoptosis level raised (cell apoptosis indexes were 16.27%?6.49% versus 33.39%? 7.05%, 33.42%?7.11%, respectively). There was no significant difference between group 3 and group 4. Conclusion: Both locally applied slow-releasing Sirolimus and oral Rapamune could inhibit vein graft neointima hyperplasia; Administration via local delivery was preferred for little side-effect on the whole body. This conclusion provides support for clinical application.