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Objective To study the clinical use of indocyanine green (ICG) fluorescence imaging in laparoscopic liver surgery.Methods The clinical and pathological data of 68 patients who underwent laparoscopic hepatectomy using the ICG fluorescence imaging technique during the study period from September 2016 to October 2018 in Zhongnan Hospital,Wuhan University were retrospectively analyzed.Analysis was carried out on the surgical methods,fluorescence navigation methods,ICG injection time and dose,tumor characteristics,and pathological studies of the resected specimens.Results Of 68 patients,3 patients were converted to open surgery,and the remaining 65 patients completed the ICG fluorescence laparoscopic hepatectomy.Thirty-two of these 65 patients underwent ICG fluorescent guided laparoscopic anatomical resection of lower hepatic segment / hepatic hemilivers (positive staining in 17 patients,negative staining in 15 patients),with 19 patients successfully staining with ICG(19 / 32,59.4％).Postoperative histopathology showed primary hepatic solid tumors (n=31),secondary liver tumors (n=12),hepatic cysts (n=4),hepatic hemangiomas (n =5),hepatolithiasis (n =12) and hepatic focal nodular hyperplasia (n =1).These lesions were combined with hepatitis B liver fibrosis in 29 patients.Conclusions ICG fluorescence imaging positively impacted on laparoscopic liver surgery.Proper preoperative ICG injection was helpful for the identification,localization and intraoperative surgical guidance of tumors,especially for patients with deep-seated and central tumors.As a consequence,oncological and surgical safety of laparoscopic liver surgery was improved.Targeted visualization of liver segments and surgical navigation using intraoperative ICG injections facilitated accurate and precise resection of anatomical liver segments or hemi-hepatectomies.The use of intraoperative ICG fluorescence technology for hepatic hemangioma,hepatic cyst,intrahepatic bile duct stones and other benign liver lesions,helped to improve safety of surgery.
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BACKGROUND: Myogenetic seed cells are the fundamental for construction of tissue-engineered smooth muscle, and optimizing the amplification of seed cells is the key in further clinical application. OBJECTIVE: To analyze the myogenetic potential of rabbit adipose-derived stromal cells induced with the combination of MyoD, transforming growth factor β1 and 5-azacytidine, and to investigate a new way to induce cells. METHODS: The rabbit abdominal fats were isolated and then the adipose-derived stromal cells were separated by col agenase digestion method for myogenetic induction with different methods: 5-azacytidine induction group, MyoD+transforming growth factor β1 induction group, 5-azacytidine+MyoD+transforming growth factor β1 combination induction group. The blank control group was set. The morphological characteristics of cells were observed at 1, 4, 8, 12, 16, 20, 24 and 28 days after induction, and the col agen type Ⅲ level were detected with 4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, flow cytometry and immunohistochemistry. RESULTS AND CONCLUSION: Compared with the other groups, the cel activity was higher in the combination induction group and reached peak at 16 days after induction, the number and volume of myotubes were increased at 20 days with regular arrangement, and desmin showed strong expression. Cel cycle showed the ratio of adipose-derived stromal cells in the DNA replication phase was increased in the combination induction group, the ratio of cells in the clearance period was decreased; the level of col agen type Ⅲ was increased significantly, and the difference was significant. The results indicate that 5-azacytidine combined with multiple factors can promote the differentiation of adipose-derived stromal cells into myoblasts with significant cel proliferation, which is considered as the ideal method to in vitro induction of myoblasts.
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As the progress of organ and tissue transplantation technology, pancreas and islet transplantation has been introduced to treat diabetes and became hot. However, many shortcomings and re-lated issues need to be improved concerning the islet cell sources. Bone marrow mesenchymal stem cells (BMSCs) has become the hot topic on induction of islet-like cell because it was simple to access, easy to culture and possess the function of immunosuppressian. In recent years, artificial construction of pancreatic islet cells, which uses gene recombinant and gene transfection to reforge islet-like cells, has improved the induction rate of BMSCs and increased the amount of insulin serection, and it also provide new ideas and direction for the treatment of diabetes.
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Objective To study the effect of Coxsackie virus B3 (CVB3) infection on islet cells in vitro, and to explore the mechanism of islet cells caused by CVB3. Methods Bone marrow mesenchymal stem cells( BMSCs) were separated from the bone marrow and cultured. Then they were induced to differentiate into islet-like cells using nicotinamide and mercaptoethanol. Differentiated cells were detected by morphology , special staining and RT-PCR. Observe CVB3 infection on islet cells under inverse microscope and detect the specific gene fragment by RT-PCR. Results BMSCs showed half suspended shape and gathered to form a cluster after induction. Cells became red brown by dithizone specific staining. RT-PCR also proved the existence of mRNA expressing insulin. Infected islet cells appeared typical pathological changes like shrinks, refraction decreases. RT-PCR detected the desired specific gene fragment of 299 bp in infected islet cells. Conclusion CVB3 can directly injury islet cells, and damage the function of islet cells of secreting insulin.
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Objective To investigate whether the defective interfering(DI) particles of Paramyxovirus, Tianjin strain could be used as a candidate adjuvant. Methods DI particles were separated and purified. After being identified, it was equally mixed with the inactivated poliovirus (PV) vaccine. Kunming mice were administered subcutaneously with the mixture, besides we set groups of inactivated PV vaccine containing Al(OH) 3 as an adjuvant, inactivated PV vaccine of which the dose was doubled without any adjuvant, and negative control. Sera were collected in the 14th day after immunization, and the specific antibodies of PV were detected. T/B lymphocyte stimulation indexes(SI) were counted through the lymphocyte proliferation tests. The quantities of IFN-γ/IL-4 produced by the splenocytes which were stimulated again by PV antigen were detected. Results The SI and the quantity of IFN-γof the mice in the group of inactivated PV vaccine combining with DI particles of Paramyxovirus, Tianjin strain were more than other groups. Conclusion DI particles of Paramyxovirus, Tianjin strain could enhance the immune response of inactivated poliovirus, especially the cellullar immunologic response of Th1 type.
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Objective To evaluate the biological potential of surface topography of biomimetic matrix combined PRP gels with RGD modification. Methods Surface topography of nβ-TCP/Cs/PCL matrix was made by Nd: YAG laser, tissue-engineered bone was constructed in the following ways: ADSCs were loaded to nβ-TCP/Cs/PCL matrix with PRP gels plus RGD modification (group A), ADSCs were cultured to nβ-TCP/Cs/PCL matrix with RGD modification (group B), ADSCs implantation with topography-surfaced treatment of matrix (group C), ADSCs cultivation with smooth-surfaced treatment of matrix (group D). SEM and CLM were used to observe cellular pattern, survival rate, cell activity, ALP and collagen type Ⅰ level were detected at 1, 4, 8, 12, 16, 20, 24,28 days. Runx2 and OPG expressions were assayed at different interval. Results Under observation of SEM and CLM, new tissue showed more remarkable cell proliferation with abundant ECM in group A. Compared with other groups, the survival rate in group A was significantly higher (88.16 ± 1.29, P ＜ 0.05),and the level of cell activity, ALP, collagen type Ⅰ were significantly higher (0.92± 0.13, 87.27 ± 3.08, 93.27 ± 3.91, P＜ 0.05), and remarkable expression of Runx2 and OPG was also seen. Conclusion Topography-surfaced treatment of matrix combined PRP gels with RGD modification enhances the cell proliferation and acts as a feasible osteopromotive method.
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The amino acid sequences of the NP,P, M, F,HN and L proteins of the paramyxovirus Tianjin strain were analyzed by using the bioinformatics methods. Phylogenetic analysis based on 6 structural proteins among the Tianjin strain and 25 paramyxoviruses showed that the Tianjin strain belonged to the genus Respirovirus, in the subfamily Paramyxovirinae, and was most closely related to Sendal virus (SeV). Phylogenetic analysis with 14 known SeVs showed that Tianjin strain represented a new evolutionary lineage. Similarities comparisons indicated that Tianjin strain P protein was poorly conserved, sharing only 78.7%-91.9% amino acid identity with the known SeVs, while the L protein was the most conserved, having 96.0%-98.0% amino acid identity with the known SeVs. Alignments of amino acid sequences of 6 structural proteins clearly showed that Tianjin strain possessed many unique amino acid substitutions in their protein sequences, 15 in NP, 29 in P, 6 in M, 13 in F, 18 in HN, and 29 in L. These results revealed that Tianjin strain was most likely a new genotype of SeV. The presence of unique amino acid substitutions suggests that Tianjin strain maybe has a significant difference in biological, pathological, immunological, or epidemiological characteristics from the known SeVs.
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Paramyxovirus Tianjin strain, a new genotype of Sendal virus, was isolated from the lungs of common cotton-eared marmoset that died of severe respiratory infection in the marmoset colonies. The 19.28% IgM positive rate in the young children with acute respiratory tract infection suggested a close relationship between Tianjin strain and humans. Hemagglutinin-neuraminidase (HN) is its major transmembrane glycoprotein responsible for viral attachment, penetration and release. To clear the relationship between HN structure and function of paramyxovirus Tianjin strain, rHN1, rHN2 and rHN3 overlapping the ectodomain of HN protein were expressed. Their antigenicity and hemaglutination activity, as well as cross reactivity to standard antisera against influenza virus type A, type B were analyzed. The results indicated expressed rHNs have the natural antigenicity.The segment rHN2 possesses more linear epitopes exposed on the surface of the native I-IN protein than found in segments rHN3 and rHN1. The hemagglutination activity of segment rHN3 is higher than that of segments rHN2 and rHN1, and partially dependent on the three-dimensional conformation of HN3 protein. Cross-reactivity between rHNs and standard antisera against influenza virus type A, type B suggested that rHNs might not be the best alternative as specific antigens to detect virus in clinicalserum specimens.
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Gene transfection is an important method for studying gene function. The origination and development of gene transfection are reviewed in this article. Six methods of gene transfection are introduced, among which, special attention are given to two new ones: particle bombardment and photochemistry transfection technology. Besides, the advantages and disadvantages of these methods are compared in order that more suitable methods can be chosen in researches. The application of the technology in gene research is discussed in the end of this article.
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<p><b>BACKGROUND</b>To provide a simple, specific and early serodiagnostic technique for the patients with aseptic meningitis caused by echovirus.</p><p><b>METHODS</b>An indirect enzyme-linked immunosorbent assay (ELISA) has been developed to detect echovirus-IgM and the specificity and availability of the assay were also examined.</p><p><b>RESULTS</b>In 78 cerebrospinal fluid (CSF) specimens which came from the children with aseptic meningitis, the positive rate was 17.9(14/78). In 64 CSF collected from non-aseptic meningitis (bacterial meningitis and cerebral trauma), the positive rate was 1.56(1/64). In 5 CSF specimens which were ELISA positive, the positive rate of neutralization test (NT) was 4/5, all the specimens which were ELISA negative were NT negative. In this assay there was no cross-reaction with poliovirus, Coxsackie virus B type 1-6 and A type 7. By blocking and destructive test of specific IgM, all CSF specimens with ELISA positive became negative.</p><p><b>CONCLUSIONS</b>The established indirect ELISA was specific and reliable. The te st was quick, simple and available, which is suitable for early and specific clinical diagnosis, and will be greatly significant to clinical treatment.</p>
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Adulto , Criança , Pré-Escolar , Humanos , Lactente , Recém-Nascido , Anticorpos Antivirais , Líquido Cefalorraquidiano , Infecções por Echovirus , Diagnóstico , Ensaio de Imunoadsorção Enzimática , Métodos , Imunoglobulina M , Líquido Cefalorraquidiano , Meningite Asséptica , Diagnóstico , VirologiaRESUMO
<p><b>BACKGROUND</b>To clone the type common antigen gD of human herpes simplex virus I, II (HSV-I, II), the authors constructed recombinant expression vector Pmal-c2/gD and induced to express the fusion protein MBP-gD.</p><p><b>METHODS</b>The authors extracted HSV DNA,amplified gD gene by PCR assay and directly cloned it into prokaryotic expression vector pMAL-c2, then transformed it into E.coli DH5alpha. After proved to be correct by PCR, double enzyme digestion and sequencing, the fusion protein is induced to express by IPTG and detected by both Western blot and ELISA.</p><p><b>RESULTS</b>The constructed expression vector pMAL-c2/gD can be expressed with high efficiency. The product expressed was about 35.5% of the total bacterium proteins by SDS?PAGE analysis and was found nearly 39% as soluble protein,61% as inclusion in cytoplasm.</p><p><b>CONCLUSIONS</b>The authors constructed recombinant expression vector pMAL-c2/gD, the Western blotting result showed that the recombinant protein could be identified with gD specific monoclonal antibody DL6. Therefore the protein was of natural antigenic structure of gD.</p>
