Objective To study cortactin function in cancer cell endocytosis.Methods We applied cortactin siRNA interference to MDA-Mb-231,a human breast adenocarcinoma cell line in which cortactin was over-expressed,and introduced anti-cortactin immunoreagents into the cells with BioPorter system to interfere with cortactin function in vivo.Capture-ELISA assay was used to measure transferrin uptake.We used immunoblot assay to assess the effect of cortactin knock-down and immunoflurescence microscopy to examine the effect of cortactin down-regulation on transferrin uptake.Results Interference of cortactin function in cells resulted in impairment of transferrin endocytosis.Transferrin fluorescent intensity in cytoplasm in cortactin siRNA treated-cells was significantly reduced in comparison to that of mock-treated cells.Less than 50% of cells subjected to cortactin siRNA treatment had normal transferrin uptake.Endocytosis in MDA-Mb-231 cells introduced with cortactin antibodies was impaired as well,showing a 30%～ 60% reduction in transferrin uptake.Conclusion Crtactin,an actin-binding protein,plays an essential role in cell endocytosis.

Objective To research the possible influence of operation and human chorionic gonadotropin(HCG) therapy on the development of antisperm antibody (AsAb) and serum levels of estradiol (E_2) and testosterone (T) of cryptorchidism boys after orchiopexy. Methods Forty cryptorchidism boys were studied who were divided into HCG treatment group and without HCG treatment group. Twenty boys who underwent inguinal operation were studied as the operation control group and 20 normal boys as the control group. The serum were collected for detection AsAb, E_2,and T. Results There were no significant difference in serum levels of E_2 and T among the three groups(P 0.05 ). Conclusions The level of AsAb is associated with the destroy of blood barrier and the disorder of immune modulation and endocrine. Treatment with HCG has no effect on AsAb level.

OBJECTIVETo examine the expression of matrix metalloproteinases (MMP)-2, -9, tissue inhibitor of matrix metalloproteinase (TIMP)-1 and hs-CRP, and their relationship with coronary artery in children with Kawasaki disease.

METHODSOne hundred and fifty-one children with Kawasaki disease (111 cases with coronary artery damage and 40 cases without) and 60 healthy children were enrolled. The expression of MMP-2, MMP-9 and TIMP-1 was detected using ELISA, and the hs-CRP concentration was measured using the endpoint nephelometry.

RESULTSThere were significant differences in the level of MMP-2, MMP-9 and hs-CRP between the patients with or without coronary artery damage and the healthy children (p<0.05). The levels of MMP-2, MMP-9 and hs-CRP were the highest in the cardiovascular damage group (p<0.05). There were positive correlations between MMP-2, MMP-9 and TIMP-1 in children with Kawasaki disease.

CONCLUSIONSMMP-2, MMP-9, TIMP-1 and hs-CRP may play important roles in the development of Kawasaki disease. The combined measurement of MMP-2, MMP-9 and hs-CRP may be useful in the evaluation of the severity in children with Kawasaki disease.

C-Reactive Protein ; analysis ; Child ; Child, Preschool ; Female ; Humans ; Infant ; Male ; Matrix Metalloproteinase 2 ; blood ; Matrix Metalloproteinase 9 ; blood ; Mucocutaneous Lymph Node Syndrome ; blood ; etiology ; Tissue Inhibitor of Metalloproteinase-1 ; blood

To investigate the therapeutic effect of artesunate on mouse cytomegalovirus pneumonia, the BALB/c-nu mice were infected with murine cytomegalovirus-green fluorescent protein (MCMV-GFP) by nose dropping method. The experimental protocol was approved by the Medical Laboratory Animal Ethics Committee of Guangzhou Medical University. The BALB/c-nu mice were randomly divided into five groups: control group, MCMV pneumonia group, and artesunate (60, 120, and 240 mg·kg^-1) groups. The survival rate, weights, and virus loads in lungs among the groups were observed. The degree of histopathologic changes in lungs was assessed directly by hematoxylin-eosin (HE) assay. MCMV-GFP expression was assessed by immunofluorescence. In addition, reverse transcription polymerase chain reaction (RT-PCR) analysis was performed to investigate the content of major immediate early 1 (Mie1) mRNA, and enzyme-linked immunosorbent assay (ELISA) was used to detect the changes of inflammatory factors, interleukin 10 (IL-10), IL-6, and tumor necrosis factor-α (TNF-α). Western blot analysis was used to detect the expression of the changes of nuclear factor-kappa B (NF-κB) signaling pathways in total proteins. Compared with MCMV group, artesunate (120 mg·kg^-1) significantly increased body weights of MCMV-infected nude mice over 30 days, and decreased the viral titer in lung homogenate, lung inflammation, and histological severity. Moreover, the administration of artesunate (120 mg·kg^-1) could downregulate the expression of phospho-NF-κB (p-NF-κB) p65 in the lungs of mice. The present study suggested that artesunate can protect the immunocompromised mice from MCMV-induced interstitial pneumonia via downregulating NF-κB signaling pathway, thus attenuating inflammation in the lungs.

Objective:To investigate the infection status of Yersinia in the main host animals of plague in Xiahe and Luqu counties, the Himalayan marmot plague foci of Gansu Province, and to provide a basis for exploring the epidemic status of plague in these foci. Methods:Samples of the ileocecal region and contents, pharyngeal swabs (or tongue roots), and blood of the main host animals of plague in Xiahe County and Luqu County where the plague were active in the 1950s and 1960s were collected from 2014 to 2018. The Yersinia isolation, virulence determination and F1 antibody detection were performed, respectively. Results:Totally 24 strains of Yersinia were detected in 958 samples of ileocecal region and contents with a bacterial detection rate of 2.51%, which were 13 strains of Yersinia enterocolitia (Y.e), 1 strain of Yersinia kristensenii (Y.k), 2 strains of Yersinia frederiksenii/ intermedia (Y.f/i), 6 strains of Yersinia intermedia (Y.i), 1 strain of Yersinia aldouae (Y.a) and 1 strain of Yersinia massiliensis (Y.m). Totally 19 strains of Yersinia were detected in 958 samples of pharyngeal swabs (or tongue roots), and the detection rate was 1.98%, which were 8 strains of Y.e, 1 strain of Yersinia pseudotuberculosis (Y.p), 4 strains of Y.k, 1 strain of Y.f/i, 4 strains of Y.i, and 1 strain of Yersinia ruckeri (Y.r). The virulence types of 21 strains of Y.e were ail -ystA -ystB +yadA -virF -rfbc -, ail -ystA -ystB -yadA -virF -rfbc -, respectively, accounting for 9.52% (2/21) and 90.48% (19/21), none were pathogenic. The results of F1 antibody in 1 079 serum samples were all negative. Conclusions:Yersinia are widely found in the pharynx and intestines of the main host animals of plague in Xiahe and Luqu counties, and the Y.e detected are all non-pathogenic strains. The results of this investigation can provide clues for further study on the preservation of Yersinia pestis in host animals and their living environment.

OBJECTIVEHematopoietic stem cell transplantation is currently a unique curative therapy for beta-thalassemia major. However, only 30% of patients have HLA-identical siblings to serve as donors. This study investigated the feasibility of hematopoietic stem cell transplantation from HLA mismatched related donors for beta-thalassemia major in children.

METHODSBetween November 2001 and November 2007, 10 patients with beta-thalassemia major at median ages of 4.4 years (range:1.6-9.4 years) received 11 transplantations from their haploidentical donors, either HLA mismatched sibling umbilical cord bloods (n=6) or parents marrows (n=4) or sibling marrow (n=1). The conditioning regiment included fludarabine (100 mg/m2), busulfan (16 mg/kg), cyclophosphamide (200 mg/kg) and antithymocyte globulin.

RESULTSOf the 10 patients, 6 (60%) had sustained engraftment and red blood cell transfusion independence; 2 patients showed transient engraftment but rejected the graft quickly; 1 patients had no evidence of engraftment and developed aplastic anemia; 1 patient who received two transplantations had no evidence of engraftment and developed persistent aplastic anemia. All eight engrafted patients showed grade I to III acute graft-versus-host disease (GVHD), and only one developed limited skin chronic GVHD. The probability of overall and disease-free survival was 90% and 60%, respectively, with a median follow-up duration of 57.1 months (range: 2.5 to 85.1 months).

CONCLUSIONSHaploidentical stem cell transplantation is an alternative option for children with beta-thalassemia major, particularly when a matched sibling donor is not available.

Child ; Child, Preschool ; Female ; Follow-Up Studies ; Graft vs Host Disease ; etiology ; Haplotypes ; Hematopoietic Stem Cell Transplantation ; adverse effects ; Histocompatibility Testing ; Humans ; Infant ; Male ; beta-Thalassemia ; therapy

OBJECTIVETo evaluate the effect of decoction for invigorating the kidney and improving blood circulation to thrombosis and pathology on rabbit blood stasis model.

METHODThirty rabbits were ramdomly divided into normal group, model group, high dose group, low dose group and Xue Shuan Ning group. Tissue-type plasminogen activator (t-PA), plasminogen activator inhibitor (PAI), fibrinogen (Fbg) and D-dimer (DD) were investigated after those rabbits had been treated. One rot was solected randomly from each group to observe pathological changes.

RESULTThere were significant differences in t-PA, PAI, Fbg and DD between normal group and other groups is very obvious (P < 0.01) . Between groups of high dose low dose Xue Shuan Ning and model, the statistical differeces were significant, as well as between groups of high dose, low dose and Xue Shuan Ning groups (P < 0.05). However, there was no statistical difference between high dose group and high dose group (P > 0.05). The pathological changes in model group were most serious, those in Xue Shuan Ning were less serious. There were slight pathological changes in high dose group and low dose group.

CONCLUSIONModels ware made successfully. High dose group and low dose group have stronger effect on thrombosis than Xue Shuan Ning group.

Animals ; Blood Viscosity ; drug effects ; Dose-Response Relationship, Drug ; Drug Combinations ; Drugs, Chinese Herbal ; administration & dosage ; isolation & purification ; pharmacology ; Female ; Fibrin Fibrinogen Degradation Products ; metabolism ; Fibrinogen ; metabolism ; Hematocrit ; Male ; Plants, Medicinal ; chemistry ; Plasminogen Inactivators ; blood ; Rabbits ; Random Allocation ; Thrombosis ; blood ; pathology ; Tissue Plasminogen Activator ; blood

OBJECTIVETo observe the expression and clinical implication of plasma miR-328 in patients with atrial fibrillation (AF).

METHODSFifty-eight patients with AF (AF group: 17 paroxysmal AF, 21 persistent AF, and 20 permanent AF) and 15 healthy volunteers (Control group) were included. General clinical data and related biochemical parameters were collected. Plasma miR-328 levels were detected with quantitative real-time polymerase chain reaction (qRT-PCR) analysis. The correlation between plasma miR-328 and AF risk factors was analyzed.

RESULTS(1) Compared with the control group, the expression level of plasma miR-328 was significantly elevated in AF group (fold 7.72 ± 9.32) (P < 0.05). (2) In AF group, the expression of plasma miR-328 was significantly different in different type of AF[paroxysmal AF with (1.98 ± 0.81), persistent AF with (6.57 ± 5.82) and permanent AF with (13.47 ± 12.29)] (P < 0.05), and which was increased in proportion to the duration of AF. (3) There was a positive correlation between plasma miR-328 level and left atrial diameter in the AF group (r = 0.310, P < 0.05).

CONCLUSIONmiR-328 expression is significantly increased in patients with AF, which may be involved in the atrial remodeling process of AF.

Aged ; Atrial Fibrillation ; blood ; Case-Control Studies ; Female ; Humans ; Male ; MicroRNAs ; blood ; Middle Aged

OBJECTIVETo construct eukaryotic expression plasmids containing green fluorescent protein gene and CYP19 wild-type or its variants (W39R, R264C, W39R-R264C) and to observe its expression in MCF-7 and Bcap-37 cells.

METHODSThe aromatase WT cDNA sequence was obtained by RT-PCR amplification and cloned into the eukaryotic expression vector pcDNA3.1(+). pcDNA3.1(+)-CYP19-GFP plasmid was then used as the template for site-directed mutation to create variant constructs (W39R, R264C, W39R-R264C). pcDNA3.1(+)-CYP19-GFP was transfected and expressed in MCF-7 and Bcap-37 cells.

RESULTThe construction of pcDNA3.1(+)-CYP19-GFP plasmid was confirmed by enzyme digestion and DNA sequencing. pcDNA3.1(+)-CYP19(W39R)-GFP, pcDNA3.1(+)-CYP19(R264C)-GFP, pcDNA3.1(+)- CYP19(W39R-R264C)-GFP plasmids were confirmed by DNA sequencing. The MCF-7 and Bcap-37 cells transfected with the pcDNA3.1(+)-CYP19-GFP plasmid expressed reporter gene of GFP.

CONCLUSIONThe eukaryotic expression plasmids have been constructed and expressed in MCF-7 and Bcap-37 cells successfully, which lays the foundation for the research of biological activities of CYP19 variant allozymes.

Aromatase ; genetics ; Cell Line, Tumor ; Genetic Vectors ; Green Fluorescent Proteins ; genetics ; Humans ; Mutagenesis, Site-Directed ; Plasmids ; genetics ; Recombinant Fusion Proteins ; genetics ; Transfection

OBJECTIVETo compare the clinical efficacy in the treatment of post-stroke shoulder-hand syndrome between floating-needle therapy and conventional acupuncture on the basis of rehabilitation training.

METHODSOne hundred cases of post-stroke shoulder-hand syndrome were randomized into a floating-needle group and an acupuncture group, 50 cases in each one. The passive and positive rehabilitation training was adopted in the two groups. Additionally, in the floating-needle group, the floating-needle therapy was used. The needle was inserted at the site 5 to 10 cm away from myofasical trigger point (MTrP), manipulated and scattered subcutaneously, for 2 min continuously. In the acupuncture group, the conventional acupuncture was applied at Jianqian (EX-UE), Jianyu (LI 15), Jianliao (TE 14), etc. The treatment was given once every two days, 3 times a week, and 14 days of treatment were required. The shoulder hand syndrome scale (SHSS), the short form McGill pain scale (SF-MPQ) and the modified Fugl-Meyer motor function scale (FMA) were used to evaluate the damage severity, pain and motor function of the upper limbs before and after treatment in the two groups. The clinical efficacy was compared between the two groups.

RESULTSSHSS score, SF-MPQ score and FMA score were improved significantly after treatment in the two groups (all P < 0.01), and the improvements in the floating-needle group were superior to those in the acupuncture group (all P < 0.05). The total effective rate was 94.0% (47/50) in the floating-needle group, which was better than 90.0% (45/50) in the acupuncture group (P < 0.05).

CONCLUSIONThe floating-needle therapy combined with rehabilitation training achieves a satisfactory efficacy on post-stroke shoulder-hand syndrome, which is better than the combined therapy of conventional acupuncture and rehabilitation training.

Acupuncture Therapy ; instrumentation ; methods ; Adult ; Aged ; Aged, 80 and over ; Female ; Humans ; Male ; Middle Aged ; Reflex Sympathetic Dystrophy ; etiology ; rehabilitation ; therapy ; Stroke ; complications ; Treatment Outcome ; Young Adult