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Pulmonary alveolar microlithiasis (PAM) is a rare genetic disease characterized by calcifications within the alveoli in the lung. Mutations in SLC34A2 gene, which encodes a type IIb sodiumphosphate cotransporter, are responsible for PAM, leading to the intra-alveolar accumulation of phosphate which favors the formation of microliths. A "sandstorm" appearance is the typical radiographic presentation of PAM. The hallmark of this disorder is clinical-radiological dissociation, with typical imaging findings, specific pathological findings and closely correlated specific genetic mutations. The disease has an insidious onset, runs a chronic course and the prognosis is poor. There is no effective treatment except for lung transplantation. This article summarizes the epidemiology, molecular genetics and clinical features of pulmonary alveolar calculi.
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Chinese medical culture is an important part of Chinese traditional culture and high-quality resources to cultivate and practice the core values of traditional Chinese medical culture among students.Institutions of Chinese medicine should make full use ofpeople-oriented,harmony,balance,big medicine sincere and other important concepts in Chinese medical culture,enhance the self-confidence of culture combined with the features of contemporary medical students,innovate the cultivation path of the core values of Chinese medical culture,cultivate a number of medical students who have good behavior habits and noble character literacy,thus to make outstanding contributions to the future development of Chinese medical culture.
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The WeChat public platform has profoundly influenced the propagation mode of medical ethics.Through the fitting of logistic ordinary differential equation,the analysis of two-level of propagation model and path,this paper constructed the propagation mode of medical ethics based on WeChat public platform.In practice,by means of accurate positioning,pushing quality content,the integration with new media forms,online interaction and other ways,the conversion rate of WeChat push will be enhanced,which will therefore promote the transmission efficiency of medical ethics.
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Objective To survey aesthetics cognition for smiling beauty in young people smile. Methods Smile model was established. The middle incisor width/length ratio, lateral incisor and middle incisor width ratio, difference of lateral incisor and the middle incisor gingival level, gingiva exposure, buccal corridor width and smile line radian were used as variable values to change respectively. A total of 200 young people were selected to evaluate results. The differences in index of ideal value and acceptable range between different dender groups were calculated. Results The ideal value of middle incisor width/length ratio was 0.75, and acceptable range was 0.65-0.85. The ideal value of lateral incisor and middle incisor width ratio was 0.618, and acceptable range was 0.518-0.718. The ideal value of lateral incisor and the middle incisor gingival level difference was-0.5 mm, and the acceptable range was-1-0 mm. The ideal value of gingiva exposure was 0 mm, and the acceptable range was 0-2 mm. The ideal value of buccal corridor was 0.09, and the acceptable range was 0.05-0.21. Coordinate smile was ideal smile line (value=1), and the acceptable range was 0.5-1. There were no statistically significant differences in smile esthetics of six ideal value indicators and acceptable ranges between different gender groups. Conclusion The ideal values and acceptable ranges of six indexes of quantitative criteria can be used for clinical treatment.
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The etiology of deep vein thrombosis (DVT) is still not elucidated nowadays. Based on the accordance between DVT incidence and the anemophilous pollen concentration in the air, we proposed the hypothesis that allergic reaction induced by anemophilous pollen may cause "idiopathic" DVT, and proinflammatory factors may play an important role in the thrombosis process.
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Rap1 is expressed in human umbilical vein endothelial cells (HUVECs). Rap1-GTPase activating protein (Rap1GAP), with its specific target, Rap1, has been shown to be important in the regulation of many physiological and certain pathological processes. In this study, we investigated the effect of Rap1GAP expression on endothelial cell function, or, more specifically, proliferation and migration of endothelial cells. HUVECs were transfected with pcDNA3.1 (empty vector), pcDNA3.1 containing Flag-tagged-Rap1GAP or Myc-tagged-Rap1N17. The proliferation, migration and tube formation were examined and compared among the 3 groups. Expression of Rap1, Rap1GAP, extracellular signal-regulated kinase (ERK), phospho-ERK, Akt, phosphor-Akt was detected by Western blotting. The results showed that the proliferation, migration and tube formation were significantly reduced in Rap1GAP- and Rap1N17-transfected HUVECs as compared with empty vector-transfected control. These changes were coincident with increased expression of Rap1GAP and decreased expression of activated Rap1, phospho-ERK and -Akt. After treatment of Rap1GAP-transfected HUVECs with a stimulator of Rap1 guanine-nucleotide-exchange factor (Rap1GEF) 8CPT-2'OMe-cAMP, it was found that Rap1 activity was decreased as compared with empty vector-transfected control. Pretreatment of HUVECs with an ERK inhibitor PD98059 or a PI3K inhibitor LY294002 prior to stimulation not only blocked 8CPT-2'OMe-cAMP-induced phosphorylation of ERK and Akt, but also significantly reduced cell proliferation and migration. Finally, we examined the effect of vascular endothelial growth factor (VEGF) on HUVECs overexpressing Rap1GAP. VEGF-stimulated Rap1 activity, phosphorylation of ERK and Akt, cyclin D1 expression and cell proliferation were repressed in HUVECs overexpressing Rap1GAP as compared to empty vector-transfected control. Taken together, our findings demonstrate that Rap1GAP/Rap1 and their downstream effectors regulate proliferation and migration of HUVECs via ERK and Akt pathways.
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Objective To summarize the clinical experience of hepatectomy with hepatic artery resection and reconstruction using gastroduodenal artery during radical resection of hilar cholangiocarcinoma.Methods From Dec.2004 to Dec.2008,nine cases of hilar cholangiocarcinoma with hepatic artery invasion were subjected to radical resection comhined with tumor invaded hepatic artery resection and reconstruction using gastroduodenal artery.The clinical data of these patients were reviewed.Results Nine cases underwent hilar cholangiocarcinoma radical resection with hepatic artery resection,immediate hepatic artery reconstruction using gastroduodenal artery end to end anastomosis while hepatic artery resection exceeding 1 cm.One patient underwent partial resection of the portal vein and repair using autogenous segment of great saphenous vein.Roux-en-Y hepaticojejunostomy was performed in 9 patients with intrahepatic bile duct stents in 8 patients.All patients suffered from postoperative transient SIRS and recovered within 2-3 days after operation.One patient experienced massive bleeding from the upper alimentary tract 3 day after operation and the bleeding was controlled afterwards.The blood flow in the reconstructed hepatic arteries monitored by Doppler was normal two weeks after operation.There was no inhospital mortality.9 patients were followed up for 1-4 years,the median survival time is 23 months (6 months to 32 months).Conclusion Hepatic artery can be reconstructed using gastroduodenal artery during a radical resection of hilar cholangiocarcinoma,and hepatic artery reconstruction decreases the postoperative complications.
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BACKGROUND: Research about magnetic drug microsphere has developed rapidly at home or abroad; additionally, previous studies have been focused on glucan, chitosan, and polyester as coating materials; however, gel material has been less reported,in particular gel-coating berberine microsphere has been not reported yet.OBJECTIVE: To prepare berberine magnetic gelatin drug microsphere, and to evaluate drug releasing, loading, and cross-linking using dynamic dialysis and light electron microscope.DESIGN, TIME AND SETTING: Preparation and property of microsphere were performed in Guizhou Yikang Pharmacy Co., Ltd.And Guizhou University from February 2004 to May 2006.MATERIALS: Gel, berberine, glutaraldehyde, liquid paraffin, and Fe_3O_4 were used in this study.METHODS: Magnetic microspheres of berberine drug with gelatin and Fe_3O_4 were prepared by a reverse phase suspension cold-condensation method according to biocompatibility of gel, solidification of glutaraldehyde, and disperse medium of liquid paraffin.MAIN OUTCOME MEASURES: Structure, morphology, drug-loading rate, entrapment efficiency, and in vitro releasing of drug microsphere.RESULTS: The diameter of the microsphere was under 20 urn. Scanning electron microscope demonstrated that Fe_3O_4 and berberine were compounded by gel into a round shape, and the mixture was expressed on micrometer. Ultraviolet absorption and infrared analysis suggested that berberine magnetic gelatin drug microsphere still had structure and property of raw drug. The drug-loading rate of berberine microsphere was 7.2%, the content of Fe_3O_4 was 20.8%, and the entrapment efficiency was 62.6%.the sustained-releasing ability of microsphere was well, and the releasing content counted for 83.5% of total drug within 7 hours.CONCLUSION: The berberine magnetic gelatin drug microsphere which was prepared by reverse phase suspension cold-condensation method characterizes by sample preparation, small diameter, regular morphology, and great sustained-releasing ability.
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In order to investigate the immunity of unloaded dendritic cells (DCs) derived from murine bone marrow to preexisting lung melanoma metastases of mice, MO5 were intravenously injected to induce lung metastases in syngeneic C57BL/6 mice. Unloaded GM-CSF DCs, PBS and DCs+SIINFEKEL were subcutaneously injected into the mice, which were divided as experimental group, negative control group and positive control group respectively. Monoclonal antibody was used to deplete NK or T cells separately. The immunity-inhibitory effects on the lung melanoma were observed and the corresponding effector cells were examined. It was found that in the experimental and positive groups, the regression was induced in metastatic nodules in the lungs of tumor-bearing mice,but abrogated by treatment with anti-asialo-GM1 but not anti-CD8. It was concluded that the unloaded DCs could suppress the lung melanoma metastases to some extent, which was mediated by NK cells, and could be used as a potent therapeutic agents for lung tumor.
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In order to investigate the origin of neointimal smooth muscle cells in transplant arteriosclerosis in rat aortic allograft, sex-mismatched bone marrow transplantation was performed from male Wistar rats to female Wistar rats. Four weeks after transplantation, the aortic transplant model was established by means of micro-surgery in rats. The recipients were divided into 4 groups: female Wistar-female Wistar aortic isografts, female SD-female Wistar aortic allografts, male SD-male Wistar aortic allografts, female SD-chimera Wistar aortic allografts. Eight weeks after transplantation, aortic grafts were removed at autopsy and processed for histological evaluation and immunohistochemistry. The results indicated that excessive accumulation of α-SMA-positive smooth muscle cells resulted in significant neointima formation and vascular lumen stricture in rat aortic allografts.Neointima assay revealed that the neointimal area and NIA/MA ratio of transplanted artery were significantly increased in all of aortic allograft groups as compared with those in aortic isograft group (P＜0.01). Neointimal smooth muscle cells were harvested from cryostat sections of aortic allograft by microdissection method. The Sry gene-specific PCR was performed, and the result showed that a distinct DNA band of 225 bp emerged in the male-male aortic allograft group and chimera aortic allograft group respectively, but not in the female-female aortic allograft group. It was suggested that recipient bone-marrow cells, as the origin of neointimal smooth muscle cells, contributed to the pathological neointimal hyperplasia of aortic allograft and transplant arteriosclerosis.
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In order to investigate the origin of neointimal smooth muscle cells in transplant arteriosclerosis in rat aortic allograft, sex-mismatched bone marrow transplantation was performed from male Wistar rats to female Wistar rats. Four weeks after transplantation, the aortic transplant model was established by means of micro-surgery in rats. The recipients were divided into 4 groups: female Wistar-female Wistar aortic isografts, female SD female Wistar aortic allografts, male SD-male Wistar aortic allografts, female SD-chimera Wistar aortic allografts. Eight weeks after transplantation, aortic grafts were removed at autopsy and processed for histological evaluation and immunohistochemistry. The results indicated that excessive accumulation of alpha-SMA-positive smooth muscle cells resulted in significant neointima formation and vascular lumen stricture in rat aortic allografts. Neointima assay revealed that the neointimal area and NIA/MA ratio of transplanted artery were significantly increased in all of aortic allograft groups as compared with those in aortic isograft group (P<0.01). Neointimal smooth muscle cells were harvested from cryostat sections of aortic allograft by microdissection method. The Sry gene-specific PCR was performed, and the result showed that a distinct DNA band of 225 bp emerged in the male-male aortic allograft group and chimera aortic allograft group respectively, but not in the female-female aortic allograft group. It was suggested that recipient bone-marrow cells, as the origin of neointimal smooth muscle cells, contributed to the pathological neointimal hyperplasia of aortic allograft and transplant arteriosclerosis.
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The eukaryotic expression of human arresten geneand its effect on the proliferation of in vitro cultured vascular smooth cells (VSMCs) in vitro were investigated. COS-7 cells were transfected with recombinant eukaryotic expression plasmid pSecTag2-AT or control plasmid pSecTag2 mediated by liposome. Forty-eight h after transfection, reverse transcription-polymerase chain reaction (RT-PCR) was used to detect the expression of arresten mRNA in the cells,while Western blot assay was applied to detect the expression of arresten protein in concentrated supernatant. Primary VSMCs from thoracic aorta of male Sprague-Dawley rats were cultured using the tissue explant method, and identified by immunohistochemical staining with a smooth muscle-specific anti-αactin monoclonal antibody before serial subcultivation. VSMCs were then co-cultured with the concentrated supernatant and their proliferation was detected using Cell Counting Kit-8 (CCK-8) in vitro. The results showed that RT-PCR revealed that the genome of arresten-transfected cells contained a 449 bp specific fragment of arresten gene, suggesting the successful transfection. Successful protein expression in supernatants was confirmed by Western blot. CCK-8 assay showed that the proliferation of VSMCs were inhibited significantly by arresten protein as compared with control cells (F=40.154, P＜0.01). It was concluded that arresten protein expressed in eukaryotic cells can inhibit proliferation of VSMCs effectively in vitro, which would provide possibility to the animal experiments.
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The eukaryotic expression of human arresten gene and its effect on the proliferation of in vitro cultured vascular smooth cells (VSMCs) in vitro were investigated. COS-7 cells were transfected with recombinant eukaryotic expression plasmid pSecTag2-AT or control plasmid pSecTag2 mediated by liposome. Forty-eight h after transfection, reverse transcription-polymerase chain reaction (RT-PCR) was used to detect the expression of arresten mRNA in the cells, while Western blot assay was applied to detect the expression of arresten protein in concentrated supernatant. Primary VSMCs from thoracic aorta of male Sprague-Dawley rats were cultured using the tissue explant method, and identified by immunohistochemical staining with a smooth muscle-specific anti-alpha-actin monoclonal antibody before serial subcultivation. VSMCs were then co-cultured with the concentrated supernatant and their proliferation was detected using Cell Counting Kit-8 (CCK-8) in vitro. The results showed that RT-PCR revealed that the genome of arresten-transfected cells contained a 449 bp specific fragment of arresten gene, suggesting the successful transfection. Successful protein expression in supernatants was confirmed by Western blot. CCK-8 assay showed that the proliferation of VSMCs were inhibited significantly by arresten protein as compared with control cells (F=40.154, P<0.01). It was concluded that arresten protein expressed in eukaryotic cells can inhibit proliferation of VSMCs effectively in vitro, which would provide possibility to the animal experiments.
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To express recombinant arresten in Escherichia coli (E. Coli) and investigate its biological activity, prokaryotic expression vector of human arresten gene was constructed by gene engineering. Human arresten gene was amplified from recombinant plasmid pGEMArr by polymerase chain reaction (PCR), and inserted into prokaryotic expression vector pRSET containing T7 promoter. Restriction analysis and DNA sequencing verified that the arresten gene was correctly cloned into the expression vector. The recombinant plasmid pRSETAt was subsequently transformed into E. Coli BL21 (DE3), and the target gene was expressed under induction of IPTG. SDS-PAGE analysis revealed that the recombinant protein with a molecular weight of 29 kD (1 kD=0. 992 1 ku) amounted to 29 % of the total bacterial proteins. After purification and renaturation, the recombinant protein could significantly suppress the proliferation of human umbilical vein endothelial cells (HUVECs). These results suggested that the expression of a biologically active form of human arresten in the pRSET expression system laid a foundation for further study on the mechanistic insight into arresten action on angiogenesis and the development of powerful anti-cancer drugs.
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AIM: To express human Arresten gene in eukaryotic cell,and to investigate its effect on the proliferation and migration in vitro of rat primary cultured thoracic aortic vascular smooth cells (VSMCs).METHODS: COS-7 cells were transfected with recombinant eukaryotic expression plasmid pSecTag2-AT or control plasmid pSecTag2 mediated by liposome.48 hours after transfection,polymerase chain reaction(RT-PCR) was used to detect the expression of Arresten mRNA in the cells,while Western blotting assay was applied to detect expressed Arresten protein in concentrated supernatants.VSMCs were then co-cultured with the concentrated supernatants;and its proliferation was detected using cell counting kit-8(CCK-8) in vitro.Migration of VSMCs was assayed by a microchemotaxis chamber and a polycarbonate filter (Transwell's chamber) with pores of 8 ?m in diameter.RESULTS: RT-PCR revealed that the genome of Arresten-transferred cells contained a 449bp specific fragment of Arresten gene.Successful protein expression in supernatants was confirmed by Western blotting.CCK-8 assay showed that the proliferation of VSMCs was inhibited significantly by Arresten protein as compared with control group(P