Aim The purpose of this study was to compare the differential expression of 15-lipoxygenase isoenzymes in the pulmonary arteries between normoxia and hypoxia and to explore their roles in the formation of hypoxic pulmomary vasoconstriction. Method Eighteen SD rats were randomly divided into two groups(n=9):the normoxic control group breathing fresh gas and the hypoxic group breeding in animal hypoxic incubator.Immunohistochemical method,in situ hybridization and Western blot were employed to determine certain 15-lipoxygenase isoenzymes which involved in the process of hypoxic pulmonary vasoconstriction.Results ①In normoxic control group,the expression of 15-LO-1 protein was detected in the pulmonary arteries;but the expression of 15-LO-2 protein wasn’t detected.②The expression of 15-LO-1 protein in hypoxic group was much stronger than that in normoxic group (P

OBJECTIVE To investigate the autophagic effect of the compounds from the Chinese medicinal herbs, Radix polygalae as a potential neuroprotective agent that enhances the clearance of mutant Huntingtin and α- synuclein in PC- 12 cells. METHODS Radix polygalae was extracted with 75% ethanol using refluxing method, and its quality was assayed by UHPLC-TOF-MS. The autophagic effect of Radix polygalae extract, and its major components including polygalacic acid, senegenin and onjisaponin B were investigated using the green fluorescent protein-light chain 3 (GFP-LC3) autophagy detection and Western blot platforms for detecting the autophagic markers, GFP-LC3 puncta formation and LC3 II expression. The degradation of A53T α- synuclein and the inhibition ofα-synuclein oligo merization related to the Parkinson disease were assayed using Western blot and flow cytometer analysis, respectively. While the cytotoxicity and the degradation of the mutant Huntingtin associated with the Huntingtin disease were investigated using MTT method and flow cytometer analysis. RESULTS Radix polygalae ethanol extract and onjisaponin B improved the GFP-LC3 puncta formation and expression of LC3Ⅱ with time and dose manner, and this induction was activated via AMPK-mTOR and Atg 7 gene pathway. Furthermore, the clearance of α-synuclein and mutant Huntingtin was enhanced via autophagy induction with the treatment of Radix polygalae ethanol extract and onjisaponin B. CONCLUSION Findings in the current study provide detailed insights into the protective mechanism of a novel autophagy inducer, onjisaponin B, which is valuable for further investigation as a new candidate agent for modulating neurodegenerative disorders through the reduction of toxicity and clearance of mutant proteins in the cellular level.

This study was designed to investigate the correlation between autophagy and polarization of macrophages in atherosclerosis (AS) plaque in arteriosclerosis obliterans amputees. Femoral artery specimens from arteriosclerosis obliterans amputees were performed hematoxylin and eosin (HE) staining, oil red O and immunofluorescence staining to observe the morphology of atherosclerotic plaque, phenotype of macrophages and autophagy in plaque; using real-time quantitative RT-PCR technology to detect the mRNA level of M1 and M2 type markers in arterial tissue; to analyze polarized signal pathway and autophagy protein levels in macrophages by Western blotting. Arterial specimens staining showed obvious lipid deposition and obvious infiltration of amount of foam cells and inflammatory cells. Macrophages were mainly expression M1 type in percentage in fibrous plaque. Although both M1 and M2 macrophages were upregulated in atheromatous plaque, the increase was dominant in M2 type in percentage. The level of autophagy was significantly higher in the atheromatous plaque than that of fibrous plaque. The expression of tumor necrosis factor- α (TNF-α), monocyte chemotactic protein-1 (MCP-1), inducible nitric oxide synthase (iNOS), interleukin-6 (IL-6) and interleukin-12 (IL-12) mRNA was significantly higher in fibrous plaque than that of atheromatous plaque (P < 0.01 or 0.05), and arginase-1 (Arg-1), transforming growth factor-β (TGF-β), CD163 and interleukin-10 (IL-10) mRNA was significantly lower than that in atheromatous plaque (P < 0.01). The levels of p-STAT1 and NF-κB were significantly increased in fibrous plaque (P < 0.01), while p-STAT6 expression was significantly increased in atheromatous plaque (P < 0.01). The level of LC3-II was significantly higher in atheromatous plaque than that in fibrous plaque (P < 0.01). Macrophages in early atherosclerotic plaque were induced to M1 type through p-STAT1/NF-κB pathway and expressed moderate levels of autophagy; while macrophages in advanced plaques were induced to polarization of M2 type through p-STAT6 pathway. M2 macrophages expressed a higher level of autophagy than M1 macrophages.
Amputees ; Arginase ; metabolism ; Arteriosclerosis Obliterans ; pathology ; Atherosclerosis ; pathology ; Autophagy ; Cell Polarity ; Chemokine CCL2 ; metabolism ; Foam Cells ; cytology ; Humans ; Interleukin-10 ; metabolism ; Interleukin-12 ; metabolism ; Interleukin-6 ; metabolism ; Macrophages ; cytology ; NF-kappa B ; metabolism ; Nitric Oxide Synthase Type II ; metabolism ; Phenotype ; STAT6 Transcription Factor ; metabolism ; Tumor Necrosis Factor-alpha ; metabolism ; Up-Regulation

Objective To investigate the expression levels of serum vascular endothelial growth factor (VEGF), lactate dehydrogenase (LDH), sugar chain antigen-125 (CA125), and β2-microglobulin (β2-MG) in peripheral blood of patients with diffuse large B-cell lymphoma (DLBCL) and their clinical significances. Methods Thirty cases of DLBCL diagnosed by pathology in Fenyang Hospital of Shanxi Province and Shanxi Dayi Hospital from December 2011 to June 2013, 20 cases of healthy individuals as normal control group were enrolled. The levels of serum VEGF, CA125 and β2-MG in peripheral blood were detected by enzyme-linked immunosorbent assay (ELISA). Serum levels of LDH were detected by the rate method for measuring. Results The expression levels of VEGF, LDH, CA125 and β2-MG in DLBCL patients were higher than those in the healthy control group [(368±194) vs. (156±48) pg/ml, t=5.718, P=0.000;(487±252) vs. (177±32) U/L, t= 6.658, P= 0.000; (58 ±16) vs. (19 ±10) U/ml, t= 9.701, P= 0.000; (3.1 ±1.5) vs. (1.6 ±0.3 ) mg/L, t=5.612, P=0.000]. The expression levels of serum VEGF and LDH in DLBCL patients with stage Ⅲ-Ⅳ were significantly higher than those in patients with stage Ⅰ-Ⅱ [(506±165) vs. (275±154) pg/ml, t= 3.896, P=0.000; (886 ±433) vs. (220 ±86) U/L, t= 5.244, P= 0.000]. The expression levels of VEGF and LDH in DLBCL patients with bone marrow infiltration were higher than those in patients without bone marrow infiltration [(505±201) vs. (299±152) pg/ml, t= 3.148, P= 0.004; (798±463) vs. (331±166) U/L, t= 3.113, P=0.005]. There were no significant differences in the expression levels of VEGF and LDH between patients with A symptoms and B symptoms (all P>0.05). The serum levels of CA125 and β2-MG in the observation group had not relationship with clinical stage, the presence of A or B symptoms and the presence of bone marrow infiltration (all P> 0.05). The high expression of VEGF had correlation with the high expression of LDH in the observation group (r=0.458, P<0.05). Conclusions The expression levels of VEGF, LDH, CA125 andβ2-MG in DLBCL patients before treatment are high, and the high expression levels of VEGF and LDH are closely related to the clinical stage, disease progression and invasion. Combined detection of VEGF and LDH may be a useful predictor of bone marrow involvement in patients with DLBCL.

OBJECTIVETo investigate the feasibility and safety of the treatment for thoracolumbar fractures with transpedicular intracorporeal hydroxyapatite grafting and pedicle screw fixation via paraspinal approach.

METHODSFrom June 2007 to December 2008, 19 cases of thoracolumbar fractures were treated with transpedicular intracorporeal hydroxyapatite grafting and pedicle screw fixation via paraspinal approach. There were 7 female and 12 male, ranging from 21 to 57 years of age (mean 40.8 years) at surgery. The time from injury to surgery varied from 1 d to 5 d (mean 2.9 d). Nineteen patients all suffered from single thoracolumbar fracture with the distribution of injury level being T(11) in 1, T(12) in 5, L(1) in 9, and L(2) in 4. According to Denis fracture classification, there were 5 compression fractures and 14 burst fractures. The mean preoperative ratio of the anterior height of the body was 57.2%, kyphosis angle was 17.6° and occupation of spinal canal was 27.7%. The mean preoperative load-sharing classification of spine fractures was 5.2. Based on the ASIA neurologic grading system, preoperative neurological function was grade B in 2 cases, C in 9 and D in 8.

RESULTSMedian operating time was 83.8 min (range 60-95 min) and median blood loss was 133 ml (range 90 - 200 ml). Infection did not occur in any of the patients and the operative incisions were healing well. Average follow-up time was 19.2 months (range 12 - 36 months). At the latest follow-up, the height of the anterior border was corrected to 88.4%, the kyphosis angle was 6.1°, and the occupation of spinal canal was 8.2% on average. The postoperative neurologic function of all 19 patients was improved with grade D in 2 cases and E in 17. There were no instances of instrumentation failure and no patient had persistent postoperative back pain.

CONCLUSIONSTranspedicular intracorporeal hydroxyapatite grafting and pedicle screw fixation via paraspinal approach could provide reliable neurologic improvement in patients with incomplete neurologic deficit, and could prevent the development of kyphosis.Furthermore, it has the obvious advantages of less invasive and blood loss, and decreases the risks of postoperative lumbodorsal pain.

Adult ; Bone Screws ; Bone Transplantation ; methods ; Durapatite ; Feasibility Studies ; Female ; Follow-Up Studies ; Fracture Fixation, Internal ; methods ; Humans ; Lumbar Vertebrae ; injuries ; Male ; Middle Aged ; Spinal Fractures ; surgery ; Thoracic Vertebrae ; injuries ; Treatment Outcome ; Young Adult

Objective To assess the neointimal coverage after the implantation of various drug eluting stents (DES) by optical coherence tomography ( OCT). Methods The study comprised of 62 patients implanted DES for ( 15. 3 ± 5.7 ) months. Patients were divided into three groups according to the type of implanted stent: Cypher group ( patient = 26, stent = 57 ), Endeavor group ( patient = 17, stent = 23 )and Firebird group (patient = 19, stent = 32). OCT images of the stent were analyzed by software equipped by Light Lab system. Intimal thickness of 64 μm, 168 μm and 366 μm represents 10%, 25% and 50%lumen area loss, respectively. Neointimal coverage was thin with intimal thickness≤64 μm, satisfactory with intimal thickness between 65 μm and 366 μm and hyperplaisa and restenosis with intimal thickness ＞ 366μm. Results The percent of complete neointimal coverage was similar among groups ( P ＞ 0. 05 ). The thickness of neointimal coverage in Cypher and Endeavor and Firebird group was (178.7 ± 11.9)μm,(228.7 ± 17. 1 ) μm and ( 170. 3 ± 13.3 ) μm, respectively( all P ＜ 0. 05 ). The symmetry of Cypher stent was better than Firebird stent, and the symmetry of Firebird stent was better than Endeavoe stent. Conclusion There was significant difference on neointimal coverage after various types of DES implantation, and OCT can be used to evaluate the symmetry of neointimal coverage post implantation of various DES.

OBJECTIVETo study the ABO allele molecular characteristics of Ael blood subgroup.

METHODSFive individuals of diagnosed as Ael blood subgroup were subjected to PCR amplify ABO alleles using four pairs of sequence-specific primers. Exon 6 and exon 7 at ABO locus of all samples were sequenced. An individual with AelB phenotype was chosen for further analysis of transcript structure of ABO gene.

RESULTSSequence analysis indicated one Ael phenotype sample with reported Ael01 allele, one Ael phenotype sample with an Ael05 allele, and two AelB and one Ael individuals did not contain referred A allele, but contain O01 or O02 allele with 261G deletion.

CONCLUSIONMolecular bases for the Ael have highly polymorphism. The mechanism responsible for the express weak A antigen of O allele with 261G deletion awaits to be elucidated.

ABO Blood-Group System ; genetics ; Alleles ; Base Sequence ; Cloning, Molecular ; DNA ; analysis ; Female ; Humans ; Male ; Molecular Sequence Data ; Polymerase Chain Reaction ; Polymorphism, Genetic

OBJECTIVETo study the effect of macrocalyxin A (MA) on proliferation, differentiation and apoptosis in HL-60 cells and explore its possible mechanisms.

METHODSDifferent concentration of MA were used to treat HL-60 cells. Proliferation inhibition was analyzed by Trypan blue staining and MTT assay, cell apoptosis by cell morphology, DNA content, cell cycle analysis, Annexin-V/PI and Hoechst 33258 fluorescence staining. The differentiation of HL-60 cells was evaluated by cell morphology, NBT tests and expression of CD11b, CD13, CD14. The expressions of bcl-2, bax, Fas, P53, mitochondrial transmembrane-potential (DeltaPsim) and mitochondrial membrane protein were analyzed by flow cytometry.

RESULTSMA could inhibit HL-60 cells proliferation capacity in a time-and dose-effect, with a 24 h IC50 value of 8.76 microg/ml, 48 h of 7.17 microg/ml and 72 h of 7.14 microg/ml. The HL-60 cells apoptosis was confirmed by cell morphology, sub-G1 phase and Annexin-V/PI labeling in a time and dose dependent manner. The more mature HL-60 cells were a with higher positivity of NBT and expressions of CD11b than those cultured without MA. The expression of bax was increased, while bcl-2, P53, Fas were unchanged on the MA treatment. MA could increase the expression of mitochondrial membrane protein in a dose-dependent manner while the DeltaPsim was reduced.

CONCLUSIONMA can inhibit proliferation and induce differentiation and apoptosis of the HL-60 cells. The mechanism may be related with up-regulating bax, opening the mitochondrial permeability transition pore and reducing DeltaPsim.

Apoptosis ; drug effects ; Cell Differentiation ; drug effects ; Cell Proliferation ; drug effects ; Diterpenes ; pharmacology ; HL-60 Cells ; Humans

OBJECTIVETo observe the therapeutic effect of needle-pricking therapy combined with spinal massage on ankylosing spondylitis and to probe into the mechanism.

METHODSNinety-three cases who were definitely diagnosed as having ankylosing spondylitis at active stage were randomly divided into a medication group (n=46) and an observation group (n=47). The observation group were treated by needle-pricking the main points, Neck No. 2 nerve, Neck No. 5 nerve point, etc., combined with spinal rotation massage, and the medication group were treated with Azulfidine. Changes of cumulative score of arthralgia and arthroncus, function of joint, Keitel test, and ESR and CRP before and after treatment were observed.

RESULTSThe effective rate and the markedly effective rate were 95.8%0 and 68.1% in the observation group and 78.3% and 23.9% in the medication group, respectively, the former being significantly better than the latter (P < 0.01).

CONCLUSIONNeedle-pricking therapy combined with spinal massage has a significant therapeutic effect with a steady and long-term effect for treatment of ankylosing spondylitis at active stage.

Acupuncture Therapy ; methods ; Adolescent ; Adult ; Child ; Combined Modality Therapy ; Female ; Humans ; Male ; Massage ; Medicine, Chinese Traditional ; Middle Aged ; Spine ; Spondylitis, Ankylosing ; therapy

OBJECTIVEThis is a study on some ABO subgroup samples which show discordant results of serological and molecular blood typing, the aim is to clarify their true ABO type by means of nucleotide analysis on exons 6 and 7 of their ABO gene.

METHODSAbsorb-elution test and family investigation were conducted to study 7 samples which were involved in ABO grouping discrepancies. Duplex polymerase chain reaction-restriction fragment length polymorphism(PCR-RFLP) method was used to identify their ABO genotypes. PCR products of exons 6 and 7 were cloned and sequenced.

RESULTSAll the 7 ABO subgroup samples with the discordant results of serological and molecular blood typing were found to have the normal O gene. Four out of them were typed as ABsub by serology, they were all of the A*102/O genotype. Sequencing analysis found all their A gene having the nt467 (C-->T) and nt803 (G-->C) mutation by comparison with the A*101 allele, i.e. their real type should be CisAB/O. Three out of 7 were typed as AsubB by serology and as BO by genotype; and point mutation was detected in all of their B gene. One of them had the nt700 (C-->G) mutation, the other 2 unrelated individuals had the novel nt640 (A-->G) mutation in their B alleles.

CONCLUSIONThrough nucleotide analysis, 7 samples have been typed as AB subgroup in serology with the normal O gene, their real ABO type being CisAB in 4 cases and B(A) in 3 cases. At the same time, a kind of novel B (A)640 allele has been uncovered in this study.

ABO Blood-Group System ; genetics ; Asian Continental Ancestry Group ; genetics ; Blood Grouping and Crossmatching ; China ; Female ; Genotype ; Humans ; Male ; Mutation ; Polymerase Chain Reaction ; Polymorphism, Restriction Fragment Length