摘要
@#Objective - To analyze the relationship between cobalt level of post shift urine and individual exposure level of , cobalt and its compounds in cobalt exposed workers and to explore the feasibility of using urine cobalt as a biomarker. Methods - A total of 148 occupational cobalt exposed workers from a new material company were selected as the exposed , - - group and 44 non occupational cobalt exposed workers from the company were selected as the control group using the typical sampling method. The exposure concentration of time weighted average of cobalt and its compounds in the workplace air of the - two groups was determined by inductively coupled plasma mass spectrometry as the individual exposure level. The cobalt levels - - of pre shift and post shift urinary samples of the two groups were detected by this method. The linear relationship between the - cobalt level of post shift urine and the individual exposure level of cobalt and its compounds in the air of the workplace was Results - 3 analyzed. The individual exposure level of cobalt and its compounds in the exposed group was 1.10 131.71 μg/m with (M) 3 the median of 12.23 μg/m. No cobalt and its compounds were detected in the workplace air in the control group. The cobalt - - levels of pre shift and post shift urines in exposed group were higher than those in the control group at the same time point (M: vs , vs , P ) - - 1.54 0.56 μg/L 8.77 0.83 μg/L all <0.01 . The cobalt level of post shift urine was higher than that in pre shift (M: vs ,P ), urine in the exposed group 8.77 1.54 μg/L <0.01 and it was positively correlated with the individual exposure level ( ,P ) , of cobalt and its compounds Spearman correlation coefficient=0.86 <0.01 . After common logarithm conversion the linear regression equation of the cobalt level of post shift urine and the common logarithm of individual exposure level of cobalt and (x) :ŷ x( ;F , its compounds in the exposed group was as follows = −0.178 + 0.988 coefficient of determination=0.72 =374.75 P ;t , P ) Conclusion - <0.01 = - 19.36 <0.01 . There was a linear correlation between cobalt level of post shift urine and occupational cobalt exposure level of cobalt exposed workers. Urinary cobalt can be used as a biomarker of occupational cobalt
摘要
OBJECTIVE To investigate the pharmacological effect and mechanism of Sanguisorba officinalis L. (SOL) in non-small cell lung cancer (NSCLC) in vitro and in vivo based on network pharmacology. METHODS Network pharmacology was used to analyze the improving effect of SOL on NSCLC and possible targets. Cell counting kit 8 (CCK-8) and 5-ethynyl-2'-deoxyuridine (EdU) staining, Western blotting, flow cytometry of Annexin Ⅴ/PI, Hoechst 33342/PI staining detection and immunofluorescence were utilized in vitro. H&E staining, immunohistochemistry staining and Western blotting were performed in vivo. RESULTS Based on network prediction, we analyzed the 208 common targets of SOL and NSCLC. 36 core targets in 208 common targets were obtained through cytoscape analysis. And the top 10 core targets included Akt, mTOR, EGFR, etc.. KEGG analysis showed that PI3K-Akt signaling pathway was the most likely pathway. Furthermore, the mechanism study found that SOL could activate the PI3K/Akt/mTOR signaling pathway in vitro and in vivo. The anti-proliferative effect of SOL in A549 and H1299 cells was measured and validated by CCK-8 and EdU assay. Immunohistochemical results of Ki67 showed that SOL effectively inhibited tumor growth in vivo. SOL also significantly inhibited the migration and invasion of A549 and H1299 cells. SOL significantly increased the percentage of cells with PI signal in A549 and H1299, and the process of cell death of A549 cells indicated that SOL induced apoptosis. The PARP-1 and caspase-3 in A549 and H1299 were found to be activated in a dose manner. The results in vivo were consistent with those in vitro. CONCLUSION SOL-induced, caspase-3-mediated apoptosis via the induction of the PI3K/Akt/mTOR signaling pathway in NSCLC, which further clarified the mechanism of SOL in the inhibition of NSCLC, and thereby provided a possibility for SOL to serve as a novel therapeutic agent for NSCLC in the future.
摘要
<p><b>OBJECTIVE</b>To investigate the effects of total saponins from Sanguisorba officinalis (DYS) on hematopoietic cell proliferation, differentiation and the expression level of IL-3R and c-kit.</p><p><b>METHOD</b>Baf3 and 32D cells were cultured with or without IL-3, then the cells were exposed to DYS in different concentrations of 5, 10, 20, 30 and 40 mg x L(-1) for 24, 48, 72 and 96 hours separately. After that, the cell proliferation and differentiation capacity were determinated by the methods of CCK8 and Giemsa staining separately. The effects of DYS on the expression level of IL-3 receptor in Baf3 cells and the expression level of c-kit in 32D cells were determinated using RT-PCR.</p><p><b>RESULT</b>DYS promotes alone proliferation of Baf3 cells and 32D cells after 48 h. In contrast to control cells, 32D cells containing DYS without IL-3 form many large clusters. DYS also increases the proliferation when cultured with IL-3. High concentration of DYS induce alone the differentiation of 32D cells and increase alone the number of the polyploidy megakaryocyte. Moreover, DYS increases alone the expression level of IL-3R in Baf3 cells and the expression level of c-kit in 32D cells separately.</p><p><b>CONCLUSION</b>Our data shows DYS can promote alone proliferation and differentiation of megakaryocyte progenitor cells. The proliferative and differentiative effect of DYS on megakaryocyte progenitor cells is correlated to the up-regulation of IL-3 receptor and c-kit expression.</p>
Subject(s)
Animals , Mice , Cell Differentiation , Genetics , Cell Line , Cell Proliferation , Dose-Response Relationship, Drug , Gene Expression , Interleukin-3 , Pharmacology , Megakaryocyte Progenitor Cells , Metabolism , Proto-Oncogene Proteins c-kit , Genetics , Receptors, Interleukin-3 , Genetics , Reverse Transcriptase Polymerase Chain Reaction , Sanguisorba , Chemistry , Saponins , Pharmacology , Time Factors摘要
Based on the NSP4 sequence of bovine rotavirus (BRV), the shRNA was designed and synthesized, and a shRNA recombinant lenti-virus vector RNAi-H1-89 was constructed. The recombinant RNAi-H1-89 Lenti-virus was packaged by transfecting the 293T cell with the recombinant vector RNAi-H1-89 and two helper plasmids using lipofectamine, and then used to infect MA104 cells. The MA104 cells were further infected with BRV strain G6 24h post-infection, with the LacZ shRNA recombinant lenti-virus as control. Thirty-six hours later, the CPE of the infected cells was observed under microscope, shRNA of NSP4 gene inhibited CPE in MA104 cell; the shRNA against NSP4 gene also inhibited NSP4 gene expression by RT-PCR, The virus titer in the cell culture supernatant was significant lower compared with the control group. The above results showed that RNAi-H1-89 against NSP4 gene could specifically silence NSP4 gene expression, and inhibit the proliferation of BRV.
Subject(s)
Animals , Cattle , Base Sequence , Cell Line , DNA, Recombinant , Genetics , Glycoproteins , Genetics , Molecular Sequence Data , Plasmids , Genetics , RNA, Small Interfering , Genetics , Rotavirus , Genetics , Physiology , Toxins, Biological , Genetics , Viral Load , Genetics , Viral Nonstructural Proteins , Genetics , Virus Replication , Genetics摘要
<p><b>AIM</b>To investigate the sweat regulation mechanism of human body.</p><p><b>METHODS</b>Arm muscular work was performed on bicycle ergometer by eight healthy male subjects on 20 W and 40 W work loads lasting 10 min or 20 min in 16 degrees C and 21 degrees C ambient temperature. Sweat, metabolic rate and corresponding skin and core temperature changes were measured during different periods.</p><p><b>RESULTS</b>Sweat varied directly with ambient temperature and there were the corresponding changes in mean skin temperature, rectal temperature and metabolic rate. And when the work load was elevated, the skin temperature at chest and metabolic rate increased as sweat increased. There were no differences in the physiological indices between 20 W 20 min and 20 W 10 min, but mean skin temperature and sweat rate during 40 W 20 min work were higher than 40 W 10 min while metabolic rate did not change. The time when chest temperature arrived at the threshold was in correspondence with obvious sweat onset. Both local skin temperature at chest and metabolic rate were significantly correlated with sweat, but the latter was stronger. The regression equation relating metabolic rate and sweat rate was compound function.</p><p><b>CONCLUSION</b>Skin temperature was important for sweat onset, and the sweat predicted model based on the metabolic rate or ambient temperature was more suitable and practical.</p>
Subject(s)
Adult , Humans , Male , Body Temperature Regulation , Physiology , Energy Metabolism , Environment , Sweating , Physiology , Temperature摘要
<p><b>OBJECTIVE</b>To observe the effects of the grafting of a platelet-derived growth factor gene-modified artificial composite skin on rat wounds with full thickness defect.</p><p><b>METHODS</b>Platelet derived growth factor-B (PDGF-B) eukaryotic expression plasmid was constructed, and the fibroblasts were transfected with it by liposome mediation. Artificial composite skins 1 and 2 were constructed respectively. The skin1 was composed of keratinocyte, porcine acellular dermal matrix and PDGF-B gene-transfected fibroblasts while the skin 2 contained keratinocyte, porcine acellular dermal matrix and fibroblasts. The two kinds of composite skin were grafted onto wounds on the rat back to form composite skin group 1 (C1) and 2 (C2), respectively, with 18 rats in each group. Eight rats with wounds without treatment served as control (C) group. The survival rate of the composite skin was observed at 2 post-operative weeks (POWs). The rat wounds were examined grossly on 2, 4 and 6 POWs for the calculation of wound contraction rate. Wound tissue samples were harvested for histological examination.</p><p><b>RESULTS</b>(1) Up to 2 POWs, 14 grafts in C1 group survived completely, 3 with partial survival and 1 failure. In C2 group, 10 skin grafts survived completely, 4 with partial survival and 4 failures. (2) A scab was formed in the wound at 2 POW in C group. The surface of the grafted skin in C1 group was smooth, elastic, and showed good anti-friction properly, and it was better in quality compared with that in other two groups at 6 POW. (3) The wound contraction rate of the grafts in C group of rats was higher than that in C1 and C2 groups at 2, 4 and 6 POWs, while that in C1 was lower than that in C2 group. (4) Capillary formation was more intense in the grafted skins in C1 group at 2 POWs, and the epithelia differentiated well into 7 to 10 layers of epithelial cells with compact and orderly arrangement and evenly distributed fibrous tissue at 6 POWs.</p><p><b>CONCLUSION</b>Repair of the wound with artificial composite skin containing PDGF-B gene could improve the quality of wound healing.</p>
Subject(s)
Animals , Female , Humans , Rats , Cells, Cultured , Proto-Oncogene Proteins c-sis , Genetics , Rats, Sprague-Dawley , Skin Transplantation , Methods , Skin, Artificial , Swine , Tissue Engineering , Transfection , Wound Healing摘要
<p><b>OBJECTIVE</b>To introduce a compound external ear framework characterized by few complications, simplicity and very low rate of implant exposure or absorption.</p><p><b>METHODS</b>A compound auricular framework, being composed of the ninth autogenous costal cartilage as ear rim and Medpor ear base, was used in one-stage total ear reconstruction. All patients were followed up postoperatively.</p><p><b>RESULTS</b>Nine cases have been performed clinically using this compound framework and all of them achieved satisfactory efficacies. The reforged ears looked natural and achieved superior cosmesis, and none of them experienced ear rim fracture or implant exposure or distortion.</p><p><b>CONCLUSION</b>This compound auricular framework has advantages of practicability, simplicity and minimal complications. It may be an ideal framework for the reconstruction of total external ear at present.</p>
Subject(s)
Adolescent , Adult , Child , Female , Humans , Male , Biocompatible Materials , Bone Transplantation , Methods , Cartilage , Transplantation , Ear, External , Congenital Abnormalities , General Surgery , Polyethylenes , Plastic Surgery Procedures , Methods , Transplantation, Autologous摘要
<p><b>OBJECTIVE</b>To investigate the role of tumor necrosis factor alpha: TNF-alpha in the hypertrophic scar and find a valid way to treat the hypertrophic scars with gene therapy.</p><p><b>METHODS</b>TNF-alpha mRNA expression was tested with a semiquantitative reverse polymerase chain reation (RT-PCR) method in the different period of a hypertrophic scar.</p><p><b>RESULTS</b>There was a significant increase of TNF-alpha mRNA as the hypertrophic scar was getting mature (P < 0.01). However, the ratios of TNF-alpha mRNA was significantly in a lower level than the normal scar.</p><p><b>CONCLUSIONS</b>The results suggest that TNF-alpha may play an important role in normal would healing, but the hypertrophic scar may result from the shortage of the TNF-alpha. This indicates that to increase the TNF-alpha with gene therapy may be a good way to treat the hyphertrophic scar.</p>
Subject(s)
Adult , Humans , Cicatrix, Hypertrophic , Genetics , Pathology , Gene Expression , RNA, Messenger , Metabolism , Reverse Transcriptase Polymerase Chain Reaction , Tumor Necrosis Factor-alpha , Genetics摘要
<p><b>OBJECTIVE</b>To establish a new method for the preparation of porcine acellular dermal matrix.</p><p><b>METHODS</b>The antigenicity of the porcine dermis was weakened by removing epidermal and dermal cells from the porcine skin through the digestion with low-concentration trypsin and repeated freeze-thaw cycles. Split thickness porcine skin was treated with 0.05% trypsin to remove the cells from the epidermis and dermis. Repeated freeze-thaw cycles were employed to further weed out the residual cells within the dermis. The prepared acellular dermis was then examined grossly, as well as histologically, and also by immunohistochemical method.</p><p><b>RESULTS</b>No cell could be identified in the prepared porcine acellular dermal matrix. The integral basement membrane was preserved on the surface of dermal matrix with compact dermal matrix collagen structure.</p><p><b>CONCLUSION</b>Low concentration trypsinization and repeated freeze-thaw cycles seemed to be a simple and effective method for the preparation of xenogeneic acellular dermal matrix.</p>
Subject(s)
Animals , Dermis , Cell Biology , Transplantation , Extracellular Matrix , Transplantation , Freezing , Skin Transplantation , Swine , Tissue Engineering , Methods , Trypsin摘要
<p><b>OBJECTIVE</b>To study the effect and the key points in the operative procedure of the one-stage reconstruction of postburn whole auricle defect with medpor car scaffold covered with superficial temporoparietal fascia (TPF) flap.</p><p><b>METHODS</b>Medpor car scaffold was embedded under the superficial temporal (TFP) fascia. Razor-thin skin was grafted onto the surface of the fascia flap.</p><p><b>RESULTS</b>Fifteen patients with postburn whole auricle defect were treated by one-stage reconstruction with Medpor ear scaffold during the last four years. It was successful in all the patients with satisfactory appearance of the reconstructed ears.</p><p><b>CONCLUSION</b>Medpor possessed friendly biological compatibility. The reconstruction gave satisfactory results, and its advantages consisted of short operational time, easy manipulation, less injury to patients and good auricular contour.</p>
Subject(s)
Adolescent , Adult , Child , Child, Preschool , Female , Humans , Young Adult , Biocompatible Materials , Burns , General Surgery , Ear Deformities, Acquired , General Surgery , Polyethylenes , Plastic Surgery Procedures , Methods , Subcutaneous Tissue , Transplantation , Surgical Flaps , Tissue Scaffolds摘要
Objective To construct an expression system to produce the virus-like particles containing a part of the sequence of PSA mRNA, which are ribonuclease-resistant due to the encapsulation of the mRNA by bacteriophage MS2 coat proteins. Methods The PCR products of PSA cDNA fragments were cloned to TA vector pBS-T, then the targeted segments could be obtained when the pBS-T-PSA were digested by restriction endonuclease Hind Ⅲ and cloned to prokaryocytic expression vector pNCCL1. The recombinant plasmids named PNCCL1-PSA were transfected into E. Coli BL21-DE3 and induced to express with IPTG. Results The recombinant plasmids were successfully constructed. The bacteriophage MS2 coat protein which expressed in BL21 can self- assemble to form ribonuclease resistant virus-like particles and the PSA mRNA was encapsulated into virus-like particles. Conclusions The virus-like particle containing PSA mRNA can be expressed in prokaryocyte and it can be used as standard and control in detecting PSA mRNA. It provides a new, stable and ribonuclease-resistant RNA standard in RNA detection.