Warm acupuncture improves arthritic injury by down-regulating expression of skeleton proteins in rats with knee osteoarthritis / 针刺研究

OBJECTIVE: To investigate the effect of warm acupuncture on chondrocyte cytoskeleton protein Rho associa-ted protein kinase (ROCK)/ monopherine domain kinase 1 (LIMK1)/Cofilin signaling of synovial tissue of the knee-joint in knee osteoarthritis (KOA) rats, so as to explore its mechanisms underlying improvement of KOA. METHODS: One hundred-twenty SD rats (half male and half female) were randomly divided into 5 groups: normal control, model, acupuncture, moxibustion and warm acupuncture, with 24 rats in each group. The KOA model was established by injection of 4% Papain (0.25 mL/kg) into the right knee cavity on day 1, 3 and 7. Rats in the acupuncture, moxibustion and warm acupuncture groups were treated with manual acupuncture, moxibustion and warm acupuncture stimulation of "Neixiyan"(EX-LE4), "Waixiyan"(EX-LE5) and "Zusanli"(ST36), respectively for 20 min, once a day for 21 days. The volume of the right knee-joint was measured by using drainage method and its width measured using a vernier caliper. The histopathological changes of the right knee cartilage were observed after H．E. stain, and scored (0 to 14 points) with reference to Markin's methods. The expression levels of ROCK, Cofilin, phospho-Cofilin, LIMK1 and phospho-LIMK1 proteins of the right knee synovial tissue were detected by Western blot. RESULTS: After modeling, the width and the volume since day 6 of the right knee-joint and Markin score of the cartilage, as well as the expression levels of ROCK, phospho-Cofilin, and phospho-LIMK1 proteins were significantly increased in the model group in contrast to the normal control group (P0.05)．. CONCLUSION: Acupuncture, moxibustion and warm acupuncture can reduce arthritic injury in KOA rats, which is closely associated with their effects in down-regulating the expression of chondrocyte cytoskeletal proteins ROCK, phospho-Cofilin and phospho-LIMK1. The efficacy of warm acupuncture is evidently superior to that of simple acupuncture and simple moxibustion.

Characterisation of the in vitro activity of a Nitazoxanide-N-methyl-1H-benzimidazole hybrid molecule against albendazole and nitazoxanide susceptible and resistant strains of Giardia intestinalis and its in vivo giardicidal activity

BACKGROUND It was previously demonstrated that CMC-20, a nitazoxanide and N-methyl-1H-benzimidazole hybrid molecule, had higher in vitro activity against Giardia intestinalis WB strain than metronidazole and albendazole and similar to nitazoxanide. OBJETIVES To evaluate the in vitro activity of CMC-20 against G. intestinalis strains with different susceptibility/resistance to albendazole and nitazoxanide and evaluate its effect on the distribution of parasite cytoskeletal proteins and its in vivo giardicidal activity. METHODS CMC-20 activity was tested against two isolates from patients with chronic and acute giardiasis, an experimentally induced albendazole resistant strain and a nitazoxanide resistant clinical isolate. CMC-20 effect on the distribution of parasite cytoskeletal proteins was analysed by indirect immunofluorescence and its activity was evaluated in a murine model of giardiasis. FINDINGS CMC-20 showed broad activity against susceptible and resistant strains to albendazole and nitaxozanide. It affected the parasite microtubule reservoir and triggered the parasite encystation. In this process, alpha-7.2 giardin co-localised with CWP-1 protein. CMC-20 reduced the infection time and cyst load in feces of G. muris infected mice similar to albendazole. MAIN CONCLUSIONS The in vitro and in vivo giardicidal activity of CMC-20 suggests its potential use in the treatment of giardiasis.

Characterisation of the in vitro activity of a Nitazoxanide-N-methyl-1H-benzimidazole hybrid molecule against albendazole and nitazoxanide susceptible and resistant strains of Giardia intestinalis and its in vivo giardicidal activity

BACKGROUND It was previously demonstrated that CMC-20, a nitazoxanide and N-methyl-1H-benzimidazole hybrid molecule, had higher in vitro activity against Giardia intestinalis WB strain than metronidazole and albendazole and similar to nitazoxanide. OBJETIVES To evaluate the in vitro activity of CMC-20 against G. intestinalis strains with different susceptibility/resistance to albendazole and nitazoxanide and evaluate its effect on the distribution of parasite cytoskeletal proteins and its in vivo giardicidal activity. METHODS CMC-20 activity was tested against two isolates from patients with chronic and acute giardiasis, an experimentally induced albendazole resistant strain and a nitazoxanide resistant clinical isolate. CMC-20 effect on the distribution of parasite cytoskeletal proteins was analysed by indirect immunofluorescence and its activity was evaluated in a murine model of giardiasis. FINDINGS CMC-20 showed broad activity against susceptible and resistant strains to albendazole and nitaxozanide. It affected the parasite microtubule reservoir and triggered the parasite encystation. In this process, alpha-7.2 giardin co-localised with CWP-1 protein. CMC-20 reduced the infection time and cyst load in feces of G. muris infected mice similar to albendazole. MAIN CONCLUSIONS The in vitro and in vivo giardicidal activity of CMC-20 suggests its potential use in the treatment of giardiasis.

Effect of Electroacupuncture on Spermatogenesis in Rats with Oligozoospermia of Insufficiency of Shen (Kidney) Essence Syndrome / 中国结合医学杂志

OBJECTIVE@#To assess the effect of electroacupuncture (EA) on expression of cytoskeletal proteins from Sertoli cells (SCs) and spermatogenesis in rats with oligozoospermia of insufficiency of Shen (Kidney) essence syndrome (OIKES).@*METHODS@#Twenty healthy male Sprague-Dawley rats were randomly assigned to four groups using a random number table: control, tripterygium glycosides (TG) treatment, sham and EA groups (n=5 in each group). A rat model of OIKES was established by oral gavage with TG. The EA group was treated with TG and received EA at Shenshu (BL 23) and Zusanli (ST 36) acupoints for 20 min, once daily for 30 days, while the sham group received EA at identical acupoints with skin penetration without stimulation. After 30 days, the final body weight and coefficients for the testis and epididymis were calculated and sperm parameters were measured. Immunohistochemical analyses were performed to detect expression of vimentin and α-tubulin in SCs and proliferating cell nuclear antigen (PCNA) immunoreactivity in germ cells. Apoptosis in germ cells was quantified by the transferase biotin-dUTP nick end labeling assay.@*RESULTS@#Compared with the control group, the final body weight and testis/epididymis coefficients of rats in the TG-treated group were not significantly different, but the sperm count and motility were lower (P<0.05). Expressions of vimentin and α-tubulin were also significantly weaker (P<0.01). The PCNA immunoreactivity of germ cells was decreased (P=0.059), whereas the apoptotic index of germ cells was increased significantly (P<0.01). In contrast, EA at BL 23 and ST 36 acupoints significantly improved the final body weight as well as the sperm count, concentration and motility (P<0.01 or P<0.05). EA increased expression of vimentin and α-tubulin in SCs markedly, and significantly enhanced PCNA immunoreactivity with decreased apoptosis in germ cells (P<0.01 or P<0.05).@*CONCLUSIONS@#EA at BL 23 and ST 36 acupoints has protective effects on spermatogenesis in rats with OIKES. This effect seems to be achieved by attenuating TG-induced disruption of cytoskeletal protein in SCs.

The clinical value of Homer expression in peripheral blood leukocytes of patients with ischemic stroke / 中华检验医学杂志

Objective To investigate clinical significance of Homer expression in peripheral blood leukocytes of patients with ischemic stroke (IS).Methods It was a retrospecive study.The gene expression levels of Homer were measured by RT-qPCR.266 patientscollected in Zhongnan Hospital from September 2015 to June 2016were divided into 5 groups:large-artery atherosclerosis (LAA,100 cases),cardioembolism (CE,42 cases),small vessel occlusion (SVO,68 cases),stroke of other demonstrated etiology (SOE,23 cases) and stroke of undemonstrated etiology (SUE,33 cases).Meanwhile,age and sex matched 126 healthy controls were also collected.IS diagnostic criteria for cerebral infarctionwas in accordance with the guideline for acute ischemic stroke in China in 2010.The levels of Homers in subgroups were compared by Oneway ANOVA.The area under curve (AUC) and 95％ confidence interval (CI) were calculated using ROC analyses.The odds ratio (OR) and 95％ CI were calculated using the multivariate logistic regression analyses.Results The levels of Homer1 [2.01 ± 0.15] and Homer2 [1.81 ± 0.31] in LAA patients were significantly higher than othergroups [Homer1 CE:2.40 ± 0.34;SVO:2.38 ± 0.35;SOE:2.36 + 0.33;SUE:2.40 ± 0.30;control group:2.35 ± 0.28;Homer2 CE:2.09 ± 0.38;SVO:2.08 ± 0.30;SOE:2.09 ± 0.41;SUE:2.10 ± 0.34;control group:2.12 ± 0.31] (Homer1 CE:t =9.353,P＜0.001;SVO:t =9.258,P＜0.001;SOE:t =5.396,P＜0.001;SUE:t=9.644,P＜0.001;control group:t =11.882,P＜0.001;Homer2 CE:t =4.725,P＜0.001;SVO:t =5.545,P＜0.001;SOE:t=3.640,P ＜ 0.001;SUE:t =4.669,P ＜ 0.001);There was no significant difference in the expression of Homer1 (F =0.940,P =0.441) and Homer2 (F =0.336,P =0.854) between non-LAA groupsand healthy controls.There was no significant difference in the expression of Homer3among the groups (F =0.641,P =0.669).Multinomial logistic regression analyses revealed that,higher Homerl (adjusted OR =8.62,95％ CI:4.13-18.00,P＜0.001) and Homer2 (adjusted OR=2.42,95％ CI:1.75-3.36,P ＜ 0.001) levels showed significant associations with increased odds of having LAA stroke,compared with the controls.ROC curves showed that the AUC of the combination of Homer1 and Homer2 for differentiating LAA and controls was 0.896 (95％ CI:0.862-0.929,P ＜0.001) and the AUCfor differentiating LAAand non-LAA was 0.847 (95％ CI:0.800-0.894,P ＜ 0.001).Conclusion The expression of Homer1 and Homer2 in peripheral blood leukocytes could be used as novel biomarkers for LAA stroke.

Cytoskeletal proteins in the follicular wall of normal andcystic ovaries of sows

The expression of cytoskeletal proteins was evaluated immunohistochemically in 36 normal ovaries sampled from 18 sows and 44 cystic ovaries sampled from of 22 sows, was evaluated. All sows had history of reproductive problems, such as infertility or subfertility. The immunohistochemically stained area (IHCSA) was quantified through image analysis to evaluate the expression of these proteins in the follicular wall of secondary, tertiary, and cystic follicles. Cytokeratins (CK) immunoreactivity was strong in the granulosa cell layer (GC) and mild in the theca interna (TI) and externa (TE) of the normal follicles. There was severe reduction of the reaction to CK in the GC in the cystic follicles, mainly in the luteinized cysts. The immunoreactivity for vimentin was higher in the GC from normal and cystic follicles in contrast with the other follicular structures. In the luteinized cysts, the IHCSA for vimentin was significantly higher in TI and in both observed cysts, the labeling was more accentuated in TE. Immunohistochemical detection of desmin and -SMA was restricted to the TE, without differences between the normal and cystic follicles. The results of the current study show that the development of ovarian cysts in sows is associated to changes in the expression of the cytoskeletal proteins CK and vimentin.

A expressão de proteínas do citoesqueleto foi avaliada por imuno-histoquímica em ovários normais e císticos de porcas matrizes. Amostras de 36 ovários normais (18 porcas) e de 44 císticos (22 porcas) foram avaliadas. Todas as matrizes apresentaram histórico de problemas reprodutivos, como infertilidade ou subfertilidade. As áreas coradas por imuno-histoquímica (IHCSA) foram quantificadas por avaliação de imagens avaliando a expressão dessas proteínas na parede folicular de folículos secundários, terciários e císticos. A imuno-reatividade para citoqueratina (CK) foi forte na camada de células da granulosa (GC) e discreta nas tecas interna (TI) e externa (TE) dos folículos normais. Houve redução acentuada da reação de CK na CG dos folículos císticos, principalmente nos cistos luteinizados. A reação para vimentina foi mais intensa na CG dos folículos normais e císticos em comparação com outras estruturas foliculares. Nos cistos luteinizados, a IHCSA para vimentina foi significativamente maior na TI e, em ambos os cistos observados, a marcação foi mais acentuada na TE. A marcação de desmina e actina alfa de músculo liso foi restrita a TE, sem diferenças entre os folículos normais e císticos. Os resultados do presente estudo mostram que o desenvolvimento de cistos ovarianos em porcas matrizes está associado a alterações na expressão das proteínas do citoesqueleto CK e vimentina.

Inhibitory effect of radixin shRNA on retinal neovascularization induced by hyperoxia in mice / 中华实验眼科杂志

Background Retinal neovascularization is pathological basis of a variety of fundus diseases,but its pathogenesis is unclear.Studies showed that the expression level of radixin in retina is remarkably increased in retinal neovascularization-related diseases.It is presumed that silencing or down-regulating the abnormal expression of radixin is helpful for curing retinal neovascularization-related diseases.Objective This study was to investigate the inhibitory effect of radixin short hairpin RNA (shRNA) plasmid on expression of radixin gene in retina of oxygen-induced retinopathy (OIR) mice.Methods Sixty-four 7-day-old C57BL/6J mice were randomly divided into normal control group, model control group, radixin shRNA plasmid group and shRNA plasmid group by random number table.There were 16 mice in every group.OIR models were established by exposing the mice in an environment of (75±2) ％ oxygen for 5 days and then returned to the normal air in the model control group,radixin shRNA plasmid group and shRNA plasmid group,while the mice of the normal control group were fed in the normal air environment.Radixin shRNA plasmid or control shRNA plasmid at the dose of 1 μg was intravitreally injected in 12-day-old mice of the radixin shRNA plasmid group or shRNA plasmid group, respectively.Five days later, FD-2000S angiography was performed on the mice of each group and then retinal flatmounts were prepared for the observation of retinal vessels.The mice from various groups were sacrificed and retinal sections were prepared.The vascular endothelial nucleus and new blood vessels extending inner limiting membrane (ILM) were examined by hematoxylin and eosin staining;the expression of radixin in the retinas was detected using immunochemistry;the relative expression levels of radixin mRNA and protein were quantitative assayed by real-time quantitative RCR and Western blot, respectively.The use and care of the animals adhered to the Association for Research in Vision and Ophthalmology Statement for the Use of Animals in Ophthalmic and Vision Research.Results The distribution of retinal vessels was normal in the normal control group.Non-perfusion zone at the posterior pole of retina, circuity of blood vessels,leakage of vessel wall and new blood vessels were found in the mice of the model control group.Non-perfusion zone and microaneurysms were also exhibited in the shRNA plasmid group.However,these findings were slight in the radixin shRNA plasmid group.The surface of ILM was in discontinuity in the model mice and shRNA-injected mice with more vascular endothelial cell nucleus and more tubes extending ILM than that in the radixin shRNA plasmid group.The immunochemistry results showed that the expressions of radixin in the normal control group and radixin shRNA plasmid group were weaker than those in the model control group and control shRNA plasmid group.The relative expression levels of radixin mRNA were 1.002±0.043,2.236-±0.093,0.556±0.015 and 2.272±0.096 in the normal control group, model control group,radixin shRNA plasmid group and control shRNA plasmid group,and those in the radixin shRNA plasmid group were significantly reduced in comparison with the normal control group, model control group and the shRNA plasmid group (all at P＜0.01).The relative expression levels were 1.000±0.082,1.193±0.021,0.263± 0.016 and 1.235±0.005 in the normal control group,model control group,radixin shRNA plasmid group and shRNA plasmid,with the lowest expression level in the radixin shRNA plasmid group (all at P＜0.01).Conclusions Radixin shRNA can downregulate the expression of radixin gene in the retinas of OIR mice and further inhibit pathological retinal neovascularization.

Culture of bone marrow mesenchymal stem cells plus allogenic decalcified bone matrix in the knee cavity of rabbits for tissue engineered cartilage / 中国组织工程研究

10.3969/j.issn.2095-4344.2013.24.001

Study of the relationship between Wnt/β-catenin and drug resistance of choriocarcinoma / 中国医师杂志

Objective To investigate the relationship between the expression of β-catenin and drug-resistance mechanism of choriocarcinoma according to the expression of β-catenin in JEG-3 cells (human choriocarcinoma cell line) and drug resistant JEG-3/VP16 cells.Methods The mRNA and protein expressions of β-catenin were analyzed with reverse transcription-polymerase chain reaction(RT-PCR) and Western blotting.Flow cytometry was used to determine the percentages of β-catenin-positive cells in the two choriocarcinoma cell lines.Results Both drug resistant choriocarcinoma cells and drag sensitive cells were found to express β-catenin; but the expression of β-catenin mRNA (1.43 ±0.24) and protein(1.49 ±0.17)in drug resistant choriocarcinoma cells was found much higher than that in drug sensitive cells(0.65 ±0.14,0.66 ±0.16,P ＜0.01).And according to detect by flow cytometry,we found the number of β-catenin-positive cells in JEG-3/VP16 cells [(40.13 ±5.17) ％] was much more than that in JEG-3 cells [(13.15 ± 1.48) ％,P ＜ 0.01].Conclusions β-catenin was highly expressed in the drug resistant choriocarcinoma cell line (JEG-3/VP16).It indicates β-catenin might be involved in the drug resistance mechanism of choriocareinoma.

New Perspectives of Dyrk1A Role in Neurogenesis and Neuropathologic Features of Down Syndrome

Down syndrome (DS) is one of the most common genetic disorders accompanying with mental retardation, cognitive impairment, and deficits in learning and memory. The brains with DS also display many neuropathological features including alteration in neurogenesis and synaptogenesis and early onset of Alzheimer's disease (AD)-like symptoms. Triplication of all or a part of human chromosome 21, especially the 21q22.1~21q22.3 region called 'Down syndrome critical region (DSCR)', has been considered as the main cause of DS. One gene product of DSCR, dual-specificity tyrosine-phosphorylation-regulated kinase 1A (Dyrk1A), has been highlighted as a key contributor to the neural consequences of DS. This minireview summarizes accumulating recent reports about Dyrk1A involvement in the neuritogenesis, synaptogenesis, and AD-like neurofibrillary tangle formation, which is mainly focusing on Dyrk1A-mediated regulation of cytoskeletal proteins, such as tubulin, actin, and microtubule-associated protein tau. Understanding the molecular mechanisms of these phenomena may provide us a rational for new preventive and therapeutic treatment of DS.

Potential role of ezrin and its related microRNA in ovarian cancer invasion and metastasis / 中华妇产科杂志

Objective To screen microRNA (miRNA) that inhibit expression of the metastasisrelated gene ezrin in ovarian cancer cells and explore their correlation to the invasion and metastasis of ovarian cancer. Methods The differential expression of ezrin in two paired high-metastatic and lowmetastatic cell lines were examined by real time reverse transcription (RT)-PCR and western blot. A functional screen with microarray was employed to identify miRNA that were differentially expressed between SKOV3 and SKOV3ip cell lines. Three programs, TARGETSCAN ( http://www. targetscan. org ),MICROCOSM ( http ://www. ebi. ac. uk/enright-srv/microcosm/htdocs/targets/v5/) and PICTAR (http://www. pictar. mdc-berlin. de), were employed to identify all miRNA, which may inhibit the expression of ezrin and were differentially expressed between SKOV3 and SKOV3ip cells. To test the repressive potential of these miRNA, synthetic mimetics were transfected individually into SKOV3ip cells and endogenous ezrin expression levels monitored by western blot and real-time RT-PCR. Results ( 1 ) The mRNA average level of ezrin were (81.74 ± 5.34) -fold higher expression level in SKOV3ip versus SKOV3 cells ( P ＜ 0. 01 ), while (2. 61 ±0. 14)-fold in HO-8910PM versus HO-8910 cells (P ＜0. 01 ). Elevated protein level of ezrin were observed in SKOV3ip cells compared with that in SKOV3 cells, and the same that in HO-8910PM cells compared with HO-8910 cells. Paired SKOV3 and SKOV3ip cells were employed to study the more significant difference in ezrin expression between them. (2) By a functional screen using miRNA microarray combined with bioinformatics analysis,the miR-183 and miR-22 were indentified as two candidate miRNA,which may have the potential regulatory role in ezrin expression. Real time RT-PCR assays revealed that miR-183 and miR-22 were, respectively, an average of (5.84 ± 0.66)-fold and(6.67 ± 0.67)-fold higher expression level in SKOV3ip versus SKOV3 cells (P ＜0. 01 ), which were in agreement with the microarray data. A subsequent validation by western blot and real time RT-PCR revealed that over-expression of miR-183 and miR-22 could both lead to an obvious decrease in ezrin protein level,while there were not signicant difference in the level of ezrin mRNA( P ＞0. 05 ). Conclusion Increased expression of miR-183 and miR-22 may both repress the protein level of ezrin,indicating that miR-183 and miR-22 may bear a potential role in inhibiting ovarian cancer metastasis in a ezrin-mediated way.

Fibroblasts in three dimensional matrices: cell migration and matrix remodeling

Fibroblast-collagen matrix culture has facilitated the analysis of cell physiology under conditions that more closely resemble an in vivo-like environment compared to conventional 2-dimensional (2D) cell culture. Furthermore, it has led to significant progress in understanding reciprocal and adaptive interactions between fibroblasts and the collagen matrix, which occur in tissue. Recent studies on fibroblasts in 3-dimensional (3D) collagen matrices have revealed the importance of biomechanical conditions in addition to biochemical cues for cell signaling and migration. Depending on the surrounding mechanical conditions, cells utilize specific cytoskeletal proteins to adapt to their environment. More specifically, cells utilize microtubule dependent dendritic extensions to provide mechanical structure for matrix contraction under a low cell-matrix tension state, whereas cells in a high cell-matrix tension state utilize conventional acto-myosin activity for matrix remodeling. Results of collagen matrix contraction and cell migration in a 3D collagen matrix revealed that the use of appropriate growth factors led to promigratory and procontractile activity for cultured fibroblasts. Finally, the relationship between cell migration and tractional force for matrix remodeling was discussed.

Analysis on the gene mutations of MYOC in primary open angle glaucoma pedigree / 中华检验医学杂志

Objective To screen the mutations of MYOC gene in a Chinese primary open angle glaucoma (POAG) family from Cbengqing and investigate the relationship between the mutations in MYOC/TIGR gene and POAG.Methods In a large 4-generation glaucoma family, myocilin gene (MYOC) was screened in 39 family members, 8 of which were confirmed patients. Normal controls included 100 normal Chinese subjects.The known mutations of MYOC gene ( including G34C, C136T, G144T, G227A, C624G,G736A, C1009G, A1036G, C1081T, G1099A, G1138A, A1139C, T1430A, C1441A and C1442T) were detected by single strand conformation polymorphism(SSCP) , po]ymerase chain reaction restriction fragment length polymorphism (PCR-RFLP) analysis and DNA sequencing.Results G227A mutation was detected in 2 POAG patients and 1 asymptomatic patient, but not in the controls.Cl009del mutation was identified in all patients of the pedigree and an offspring member but not in the controls. No other mutations were detected.Since the C1009del mutation was revealed for the first time, a new GenBank number FJ237047 correponding to ACI62293 was applied.Conclusions The G227A mutation is a known site and there is no relationship between G227A mutation and glaucoma. But C1009del may be related to glaucoma which suggests that morbidity could be higher in the relatives of POAG than the controls.

uPA,β-catenin expression status and microvessel density correlated to invasion and metastasis in thyroid carcinoma / 中国医师杂志

Objective To investigate the expression of urokinase-type plasminogen activator(uPA)and β-catenin proteins and their relationship with tumor angiogenesis,and explore their correlation with invasion and metastasis of thyroid carcinoma.Methods SP immunohistochemistry was performed to detect the expression of uPA,β-catenin and CD105 in 90 cases of thyroid carcinoma tissues.MVD counting was performed and analyzed in conjunction with the clinicopathological features of human thyroid carcinomas.Results The positive rate of uPA expression and abnormal expression rate of β-catenin and the MVD value was positively correlated with histological typing of thymid carcinoma.There was significant difference in thyroid carcinoma with lymph node metastasis,which was significantly higher than those cases without lymph node metastasis(P＜0.05),as well as between clinical stage Ⅲ～Ⅳ and Ⅰ～Ⅱ.The MVD value was significantly correlated with the positive expression of uPA and abnormal expression of β-catenin in thyroid.There was statistically significant correlation between the expression of uPA and β-catenin.Conclusion The expression of uPA protein in thyroid carcinoma was higher than those in normal.A reduced membranous expression rate and abnormal nuclear/cytoplasmic expression of β-catenin was observed in thyroid carcinoma cells,which was positively correlated with histological typing and clinical staging,invasion and metastasis in thyroid carcinoma.uPA and β-catenin were cooperated in angiogenesis of thyroid carcinoma.MVD was positively correlated with invasion and metastasis in thyroid carcinoma.They were hopeful to be an important prognostic indicator of thyroid carcinoma.

Caracterização molecular e funcional de ANKHD1 na hematopoese normal e neoplasica / Molecular and functional characterization of ANKHD1 in normal and neoplastic hematopoiesis

A identificação e caracterização estrutural e funcional de genes diferencialmente expressos entre tecidos tumorais e normais constituem etapas fundamentais para permitir a compreensão do processo neoplásico e o desenvolvimento de novas estratégias antitumorais. Ankyrin Repeat Single KH Domain containing 1 (ANKHD1) foi inicialmente identificada em células de adenocarcinoma de próstata humano (LNCaP), no ano de 2003. Entretanto, seu padrão de expressão e sua função ainda não haviam sido caracterizados. A ANKHD1 é uma proteína ortóloga à Multiple Ankyrin repeat and single KH domain (Mask) da Drosophila melanogaster. Mask foi identificada através de um rastreamento genético utilizado para detectar novas proteínas associadas à proteína tirosina fosfatase Corkscrew (CSW), homóloga à Src Homology-2 domain-containing protein tyrosine Phosphatase-2 (SHP2) humana. SHP2 é uma fosfatase de tirosina citoplasmática codificada pelo gene PTPN11 e exerce papel fundamental no desenvolvimento da hematopoese normal e leucêmica. Os objetivos gerais do presente estudo foram caracterizar o padrão de expressão gênica de ANKHD1 em células hematopoéticas normais e leucêmicas e verificar sua função nos processos celulares. Neste estudo foi demonstrado que o gene ANKHD1localiza-se no cromossomo 5, possui vários transcritos variantes possivelmente gerados por mecanismos de clivagem alternativa e codifica proteínas com domínios de repetições de anquirina. A região promotora desse gene possui vários elementos regulatórios importantes como...

The identification and the structural and functional characterization of genes differentially expressed between tumors and normal tissues are fundamental steps towards the understanding of the neoplastic process and the development of new anti-cancer strategies. The Ankyrin Repeat Single KH Domain containing 1 (ANKHD1) was first described in humans in a prostate carcinoma cell line LNCaP, in 2003; however, the expression pattern and function of ANKHD1 have not yet been described. ANKHD1 is an orthologous protein of the Drosophila melanogaster, MASK (Multiple Ankyrin repeat and single KH domain), where it was first identified using a genetic screen designed to discover proteins that interact with the protein tyrosine phosphatase Corkscrew (CSW), which is a homolog to the SH2-containing protein tyrosine phosphatase (SHP2) in humans. SHP2 is a cytoplasmic protein-tyrosine phosphatase, coded by the PTPN11 gene and plays an important role in the development of normal hematopoiese and leukemogenesis. The aim of the present study was to characterize the gene expression pattern of ANKHD1 in normal and leukemic hematopoietic cells and to determine their function in cellular process.This study has demonstrated that the ANKHD1 gene is located on chromosome 5, this gene has several possible variant transcripts...

Advance in mechanism research of the interaction between bone plant on osteoblast / 国际生物医学工程杂志

The conformity between bone implant and bone tissue plays an important role in osseointegration.Protein molecules such as extracellular matrix proteins,cytoskeletal proteins,and attachment proteins,which produced by osteoblasts,promote the attachment of osteoblasts to bone implant and the surrounding ceils.In vitro studies show that nano-plated implant material bears excellent biocompatibility and superior physical and chemical features.Nano-scale material has the advantage of high surface roughness,wettability and electrochemistry features which erdaance the adherence and interaction of extracellular matrix proteins and promotes the attachmerit,proliferation and differentiation of osteoblasts.The expression of cytokines,such as prostaglandin E2(PGE2)and transforming growth factor-β(TGF-β)are also increased in nano-structured bone implants which in turn promote the new bone formation.Further studies need to be done to explain the mechanism of the effect of nano-tructured bone implant on osteoblasts.

Family based analysis of quantitative changes of erythrocyte membrane proteins in essential hypertension.

Our previous studies have found significant quantitative changes in the erythrocyte membrane proteins in essential hypertension (EH). The purpose of the present study was to quantify genetic and environmental contributions to quantitative variability of erythrocyte membrane proteins in EH. We studied 115 hypertensive patients, 126 normotensive subjects, 235 of their first-degree relatives and 24 twin pairs by sodium dodecyl sulphate (SDS) polyacrylamide gel electrophoresis. The decomposition of total phenotypic variance of erythrocyte membrane proteins to genetic and environmental components was performed by the least squares method. We found that genetic factors play a significant role in the control of quantitative changes in erythrocyte membrane proteins in EH. The genetic contribution to anion exchanger variation was stronger in hypertensives (88%) than in normotensives (36%), and was attributed exclusively to additive polygenic effects. Variation in glucose transporter was under marked control of major gene effect (74%). Importantly, variations in anion and glucose transporters in EH but not in healthy controls were strongly affected by common underlying genes with strong pleiotropic effects (r=0.921, P<0.05). These data provide evidence to support the genetic source of quantitative changes in membrane proteins in EH. Furthermore, the pleiotropic effects of common underlying genes seem to be responsible for variations in the transport proteins likely associated with genetic susceptibility to essential hypertension.

Expressions of E-cadherin, and ?-,?-catenin in Breast Cancers and Their Lymph-node Metastatic Tumors / 中国医师杂志

Objective To investigate the protein expressions of E-cadherin and ?-,?-catenin in breast cancers and their lymph-node metastatic tumors to evaluate their possible role in breast cancer invasion and metastasis. Methods High sensitive S-P immunohistochemical method was used to detect the protein expressions of E-cadherin and ?-,?-catenin in 60 breast cancers(31cases with lymphatic metastasis) and their lymph-node metastatic tumors. Results The abnormal expression rates of the E-cadherin and ?-,?-catenin were 51 7%, 61 7%, 70% respectively in the 60 cases of breast cancers. The expression of E-cadherin was significantly associated with lymphatic metastasis(P

Prognostic significance of E-cadherin/catenin complex expression in gastric cancer

Abnormal expression of E-cadherin/catenin complex in cancer has been associated with poor differentiation and acquisition of invasiveness, suggesting a possible role of this protein as an invasion suppressor. In this study, we conducted an immunohistochemical investigation of all components of the E-cadherin/catenin complex in 65 gastric cancer patients. Abnormal expression of E-cadherin and, alpha- and gamma-catenin occurred more frequently in diffuse than in intestinal type of gastric cancer, and correlated with poor differentiation. Abnormal expression of E-cadherin and beta-catenin correlated with poor survival. Abnormal expression of all four components of the complex was associated with poorly differentiated and diffuse-type carcinoma, and poor survival. In the multivariate analysis, abnormal expression of the E-cadherin/catenin complex was not an independent prognostic factor. These results suggest that the E-cadherin/catenin complex may be a useful marker of differentiation and prognosis in gastric cancer. Further studies are warranted to clarify the impact of the E-cadherin/catenin complex on prognostic factor of gastric cancer.

Application of the immunofluorescent technique for diagnosing muscular dystrophy / 中华神经科杂志

Objective To establish a specific detection technique in patients with Duchenne muscular dystrophy (DMD),Becker muscular dystrophy (BMD) and limb-girdle muscular dystrophy (LGMD). Methods Immunofluorescent technique was applied,and 4 monoclonal antibodies against dystrophin,?-sarcoglycan (?-SG) and ?-sarcoglycan (?-SG) were respectively used.Results Dystrophin was negative in 9 DMD patients,and partially absent in 4 BMD patients. In two LGMD patients,50DAG was diminished in one patient and 35DAG was diminished in the other. Therefore,the two patients were diagnosed as having LGMD2D and LGMD2C respectively. Conclusion Analysing gene product expressed on surface membrane of muscle fiber is helpful to diagnose and classify the DMD/BMD,and LGMD diseases by using immunofluorescent technique.