OBJECTIVETo probe therapeutic effects of centro-square needling and triple needling on ankylosing spondylitis.

METHODSSixty cases of spondylitis were randomly divided into a treatment group and a control group. The treatment group were treated with centro-square needling and triple needling, and the control group with routine acupuncture therapy, with motortherapy combined in the two groups.

RESULTSBoth the therapeutic methods were effective, but the therapeutic effect in the treatment group was significantly better than that in the control group (P < 0.05).

CONCLUSIONAcupuncture combined with motortherapy has definite therapeutic effect on ankylosing spondylitis, with centro-square needling and triple needling having better therapeutic effect.

Acupuncture Therapy ; methods ; Adult ; Exercise Therapy ; Female ; Humans ; Male ; Spondylitis, Ankylosing ; therapy

Objective To observe the effects of Yishen Tongluo Decoction (YTD) on the renal mRNA expression of transforming growth factor-β1 ( TGF-β1) and collagen Ⅳ ( ColⅣ) in membranous nephropathy ( MN) rats. Methods SD rats were randomly divided into normal group, model group, benazepril group (in the dosage of 10 mg·kg-1·d-1) , YTD group ( in the dosage of 20 g·kg-1·d-1) . The rats in various groups were given intragastric administration of corresponding agents. At the end of the fourth week, 24-hour urinary protein quantity, albumin ( ALB) , total protein ( TP) , triglyceride ( TG) , total cholesterol ( TC) , blood urea nitrogen ( BUN) , and creatinine (Cr) levels were observed. The mRNA expression levels of TGF-β1 and ColⅣ in renal tissue of rats were detected by immunofluorescence method, electron microscopy and real-time polymerase chain reaction (PCR) method. Results In the model group, urinary protein quantity in rats was increased, serum levels of TP and ALB were significantly lowered, serum levels of TC and TG were significantly increased, renal pathological changes were present, and mRNA expression levels of TGF-β1 and ColIV in renal tissue were up-regulated (P<0.05 compared with normal group). Compared with the model group, 24-hour urinary protein quantity, TC and TG levels were significantly lowered, TP and ALB levels were significantly increased, rat renal injury was relieved, mRNA expression levels of TGF-β1 and ColIV in renal tissue were down-regulated in the treatment groups ( P<0.05) . However, the differences between benazepril group and YTD group were insignificant ( P>0.05) . Conclusion The therapeutic mechanism of YTD for MN is probably related with the inhibition of mRNA expression of TGF-β1 and ColⅣin renal tissue.

BACKGROUND: Autophagy of alveolar macrophages is a crucial process in ischemia/reperfusion injury-induced acute lung injury (ALI). Bone marrow-derived mesenchymal stem cells (BM-MSCs) are multipotent cells with the potential for repairing injured sites and regulating autophagy. This study was to investigate the influence of BM-MSCs on autophagy of macrophages in the oxygen-glucose deprivation/restoration (OGD/R) microenvironment and to explore the potential mechanism. METHODS: We established a co-culture system of macrophages (RAW264.7) with BM-MSCs under OGD/R conditions in vitro. RAW264.7 cells were transfected with recombinant adenovirus (Ad-mCherry-GFP-LC3B) and autophagic status of RAW264.7 cells was observed under a fluorescence microscope. Autophagy-related proteins light chain 3 (LC3)-I, LC3-II, and p62 in RAW264.7 cells were detected by Western blotting. We used microarray expression analysis to identify the differently expressed genes between OGD/R treated macrophages and macrophages co-culture with BM-MSCs. We investigated the gene heme oxygenase-1 (HO-1), which is downstream of the phosphoinositide 3-kinase/protein kinase B (PI3K/Akt) signaling pathway. RESULTS: The ratio of LC3-II/LC3-I of OGD/R treated RAW264.7 cells was increased (1.27 ± 0.20 vs. 0.44 ± 0.08, t = 6.67, P  < 0.05), while the expression of p62 was decreased (0.77 ± 0.04 vs. 0.95 ± 0.10, t = 2.90, P  < 0.05), and PI3K (0.40 ± 0.06 vs. 0.63 ± 0.10, t = 3.42, P  < 0.05) and p-Akt/Akt ratio was also decreased (0.39 ± 0.02 vs. 0.58 ± 0.03, t = 9.13, P  < 0.05). BM-MSCs reduced the LC3-II/LC3-I ratio of OGD/R treated RAW264.7 cells (0.68 ± 0.14 vs. 1.27 ± 0.20, t = 4.12, P  < 0.05), up-regulated p62 expression (1.10 ± 0.20 vs. 0.77 ± 0.04, t = 2.80, P  < 0.05), and up-regulated PI3K (0.54 ± 0.05 vs. 0.40 ± 0.06, t = 3.11, P  < 0.05) and p-Akt/Akt ratios (0.52 ± 0.05 vs. 0.39 ± 0.02, t = 9.13, P  < 0.05). A whole-genome microarray assay screened the differentially expressed gene HO-1, which is downstream of the PI3K/Akt signaling pathway, and the alteration of HO-1 mRNA and protein expression was consistent with the data on PI3K/Akt pathway. CONCLUSIONS Our results suggest the existence of the PI3K/Akt/HO-1 signaling pathway in RAW264.7 cells under OGD/R circumstances in vitro, revealing the mechanism underlying BM-MSC-mediated regulation of autophagy and enriching the understanding of potential therapeutic targets for the treatment of ALI.
Apoptosis ; Autophagy ; Bone Marrow ; Glucose ; Heme Oxygenase-1/metabolism* ; Macrophages/metabolism* ; Mesenchymal Stem Cells/metabolism* ; Oxygen ; Phosphatidylinositol 3-Kinase ; Phosphatidylinositol 3-Kinases/metabolism* ; Proto-Oncogene Proteins c-akt/metabolism* ; Signal Transduction

BACKGROUNDTo investigate the expression of aquaporins in human pulmonary adenocarcinoma cell line SPC-A-1.

METHODSThe expressions of aquaporin 1, aquaporin 3, aquaporin 4, and aquaporin 5 in mRNA level and their locations were determined in cell line SPC-A-1 respectively by RT-PCR and immunohistochemistry.

RESULTSThe immunohistochemical stain showed aquaporin 3 and aquaporin 5 located on the membrane of SPC-A-1 cell, but no positive stain of aquaporin 1 and aquaporin 4 was observed. Both aquaporin 3 and aquaporin 5 mRNA expressed in SPC-A-1 cell line, and the expression level of aquaporin 5 mRNA was significantly higher than that of aquaporin 3 mRNA ( P < 0.01). Aquaporin 1 and aquaporin 4 mRNA did not express in SPC-A-1 cell line.

CONCLUSIONSAquaporin 3 and aquaporin 5 express in SPC-A-1 cell, and their roles in water transport of SPC-A-1 cell should be further investigated.

OBJECTIVETo compare the influence of cardiac-pulmonary function on clinical acute respiratory failure patients using Proportional assist ventilation (PAV), Pressure support ventilation (PSV) and intermittent positive pressure ventilation (IPPV). Here, we also describe some our experience with the clinical use of PAV.

METHODSUsing the IPPV mode in ten acute respiratory failure patients, calculate Elastance (Ers) and Resistance (Rrs), then change to PSV, set inspiratory positive airway pressure (IPAP) according to IPPV, so that tidal volume (V(T)) is the same as that of IPPV. We then changed the mode into PAV and set the assist ratio according to PSV, so that V(T) and Ppeak were the same as that of PSV. Then we observed the changes of respiratory mechanics, blood gas levels and hemodynamics during ventilation.

RESULTSCompared with PSV and IPPV, peak pressure (Ppeak) of PAV was markedly lower while V(T) was similar; work of breathing of patient (WOBp), and work of breathing of ventilation (WOBv) were also lower; center vein pressure (CVP) and pulmonary capillary wedge pressure (PCWP) of PAV were markedly lower than that of IPPV while V(T) were similar. Compared with PSV, V(T), mean blood pressure (mBP) and cardiac output (CO) of PAV were higher. Mean pulmonary artery pressure (mPAP) and WOBp of PAV were lower while Ppeak was similar; the differences in WOBp were notable.

CONCLUSIONSFor clinical acute respiratory failure patients, compared with PSV and IPPV, PAV has lower airway pressure, less WOBp and less influence on hemodynamics.

Acute Disease ; Adult ; Aged ; Aged, 80 and over ; Cardiac Output ; physiology ; Female ; Hemodynamics ; physiology ; Humans ; Male ; Middle Aged ; Pulmonary Ventilation ; physiology ; Pulmonary Wedge Pressure ; physiology ; Respiration, Artificial ; methods ; Respiratory Insufficiency ; physiopathology ; therapy ; Ventilators, Mechanical

Aim: To investigate the effect of triptolide combined with epidermal growth factor receptor monoclonal antibody cetuximab on the biological behavior of human colorectal cancer cell line SW480. Methods: MTT assay was used for estimating the survival rates of SW480 cells exposed to different concentrations and different durations of triptolid and cetuximab. Colony formation assay was used for showing the proliferation and wound healing assay was performed to assess the effects of drugs on cell migration, Western blot was used for testing the expression of LC3-II, p62, E-cadherin, Vimentin, mTOR, Snail, and Twist. Results: The SW480 growth of cells was inhibited by triptolide in a dose- and time-dependent manner and cetuximab was only in a dose-dependent manner. The combination regimen of cetuximab and triptolide exerted a synergistic effect. Triptolide could decrease expression of p62 increase expression of LC3-II and induce autophagic apoptosis. Triptolide monotherapy group and combination group could significantly inhibit EMT in SW480 cells, and the E-cadherin protein was up-regulated, Vimentin protein was down-regulated, and key molecules Snail and Twist were down-regulated. Conclusion: Triptolide combined with cetuximab has a synergistic effect on the inhibition of proliferation and metastasis of SW480 cells. Triptolide induces autophagic apoptosis by inhibiting the mTOR pathway. Autophagy mediated by triptolide can inhibit EMT of SW480 cells and may be a mechanism to reverse the resistance of cetuximab.

Objective: To investigate the effects of melatonin on the oxidative stress and signaling pathways of apoptosis-related genes following testicular torsion/detorsion in male rats. METHODS: Twenty-four healthy male Sprague-Dawley rats were randomly divided into a control, a torsion and a melatonin group of equal number. The torsion model was made in the animals of the latter two groups by 720° torsion of the left testis for 2 hours. The rats of the torsion and melatonin groups received intraperitoneal injection of isotonic saline and melatonin (17 mg/kg) respectively at 15 minutes prior to detorsion. At 24 hours after modeling, testis tissues were collected from the rats for detection of the apoptosis of the germ cells by flow cytometry (FCM), analysis of the expressions of Fas, Fas ligand (FasL) and Bax mRNA by quantitative real-time PCR (qRT-PCR), measurement of the cytochrome C content released from the mitochondrion by Western blot, and determination of the total antioxidant capacity (T-AOC) and the levels of myeloperoxidase (MPO) and malodialdehyde (MDA) by spectrophotometry. RESULTS: Compared with the torsion group, the rats treated with melatonin showed significantly increased normal testicular cells (［77.81 ± 6.52］% vs ［88.61 ± 7.93］%, P < 0.05), decreased early apoptotic germ cells (［16.74 ± 3.16］% vs ［6.97 ± 1.65］%, P < 0.05), down-regulated expressions of Fas (［4.52 ± 0.29］ vs ［2.66 ± 0.37］, P < 0.01), FasL (［2.82 ± 0.30］ vs ［1.73 ± 0.18］, P < 0.01) and Bax mRNA (［2.39 ± 0.18］ vs ［1.50 ± 0.14］, P < 0.01), reduced levels of cytochrome C (［1.40 ± 0.38］ vs ［0.67 ± 0.30］, P < 0.01), MPO (［0.52 ± 0.15］ vs ［0.19 ± 0.10］ U/g prot, P < 0.01) and MDA ［6.37 ± 1.73］ vs ［3.98 ± 0.90］ nmol/mg prot, P < 0.01) and elevated T-AOC (［0.76 ± 0.25］ vs ［1.55 ± 0.32］ U/mg prot, P < 0.01). CONCLUSIONS Melatonin has a significant protective effect on spermatogenesis after testicular torsion by regulating the expressions of apoptosis-related genes and increasing T-AOC in the testis tissue.

Objective To investigate the promoting effect of histone deacetylase 1 (HDAC1) expression on insulin resistance (IR) in nonalcoholic fatty liver disease (NAFLD) cells by establishing an HepG2 cell model of high fat-induced NAFLD. Methods HepG2 cells were divided into control group, model group (OA), and inhibitor group (OA+pyroxamide [an HDAC1 inhibitor]). CCK-8 assay was used to plot the standard growth curve of HepG2 cells and screen out the optimal drug concentration and action time of OA and pyroxamide; oil red O staining was used to compare the accumulation of lipid droplets in cells; an automatic biochemical analyzer was used to analyze the content of alanine aminotransferase (ALT), aspartate aminotransferase (AST), triglyceride (TG), and total cholesterol (TC) in cells; quantitative real-time PCR and Western blot were used to measure the mRNA and protein expression levels of HDAC1 and insulin receptor substrate-1 (IRS-1) in cells. A one-way analysis of variance was used for comparison of continuous data between multiple groups, and the least significant difference t -test was used for further comparison between two groups. Results OA treatment at a concentration of 0.25 mmol/L for 24 hours was the optimal concentration and duration of cell modeling, and treatment at a concentration of 20 μmol/L for 24 hours was the optimal administration concentration and duration of pyroxamide. Compared with the control group, the model group had significant increases in the content of ALT, AST, TG, and TC, and compared with the model group, the inhibitor group had significant reductions in the content of ALT, AST, TG, and TC (all P < 0.05). The model group had significantly higher mRNA and protein expression levels of HDAC1 than the control group, while the inhibitor group had significantly lower expression levels than the model group (all P < 0.05); the model group had significantly lower mRNA and protein expression levels of IRS-1 than the control group, while the inhibitor group had significantly higher expression levels than the model group (all P < 0.05). Conclusion HDAC1 participates in the development and progression of NAFLD by inhibiting the expression of IRS-1 molecule and promoting IR, and the HDAC1 inhibitor pyroxamide can exert a protective effect on the liver by alleviating IR.

BACKGROUND: There have been many significant advances in the diagnosis and treatment of non-small cell lung cancer (NSCLC). However, the mechanism underlying the progression of NSCLC is still not clear. Plant homodomain finger-like domain-containing protein 5A (PHF5A) plays an important role in processes of chromatin remodeling, morphological development of tissues and organs and maintenance of stem cell pluripotency. This study aims to investigate the role of PHF5A in the proliferation and migration of NSCLC. METHODS: A549 and PC-9 PHF5A overexpression cell lines were constructed. PHF5A expression was decreased in H292 and H1299 cells by using siRNA. Flow cytometry was used to detect the cell cycle. MTT assay and clone formation assay were used to examine the proliferative ability of NSCLC, while migration assay and wound healing assay were performed to evaluate the ability of migration. Western blot analysis was used to measure the expressions of PI3K, p-AKT and the associated downstream factors. RESULTS: Up-regulation of PHF5A in A549 and PC-9 cells increased the proliferation rate, while down-regulation of PHF5A in H292 and H1299 cells inhibited the proliferation rate at 24 h, 48 h and 72 h (P<0.05). The metastatic ability was elevated in the PHF5A-overexpresion groups, while reduced in the PHF5A-down-regulation group (P<0.05). In addition, reduced expression of PHF5A induced cell cycle arrest at G1/S phase (P<0.05). Furthermore, decreased expression of PHF5A reduced the expression levels of PI3K, phosphorylation of AKT, c-Myc (P<0.05) and elevated the expression of p21 (P<0.05). CONCLUSIONS These results demonstrated that PHF5A may play an important role in progression of NSCLC by regulating the PI3K/AKT signaling pathway.
Humans ; Carcinoma, Non-Small-Cell Lung/pathology* ; Lung Neoplasms/pathology* ; Proto-Oncogene Proteins c-akt/metabolism* ; Phosphatidylinositol 3-Kinases/metabolism* ; Cell Line, Tumor ; Cell Proliferation/genetics* ; Cell Movement/genetics* ; Gene Expression Regulation, Neoplastic ; Trans-Activators/genetics* ; RNA-Binding Proteins/metabolism*