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1.
Chinese Journal of Pathology ; (12): 1001-1005, 2023.
Artigo em Chinês | WPRIM | ID: wpr-1012353

RESUMO

Objective: To investigate the clinicopathological features of glomuvenous malformation (GVM). Methods: Thirty-one cases of GVM diagnosed at the Henan Provincial People's Hospital from January 2011 to December 2021 were collected. Their clinical and pathological features were analyzed. The expression of relevant markers was examined using immunohistochemistry. The patients were also followed up. Results: There were 16 males and 15 females in this study, with an average age of 11 years (range, 1-52 years). The locations of the disease included 13 cases in the limbs (8 cases in the upper limbs, 5 cases in the lower limbs), 9 cases in the trunks, and 9 cases in the foot (toes or subungual area). Twenty-seven of the cases were solitary and 4 were multifocal. The lesions were characterized by blue-purple papules or plaques on the skin surface, which grew slowly. The lumps became larger and appeared to be conspicuous. Microscopically, GVM mainly involved the dermis and subcutaneous tissue, with an overall ill-defined border. There were scattered or clustered irregular dilated vein-like lumens, with thin walls and various sizes. A single or multiple layers of relatively uniform cubic/glomus cells were present at the abnormal wall, with scattered small nests of the glomus cells. The endothelial cells in the wall of abnormal lumen were flat or absent. Immunohistochemistry showed that glomus cells strongly expressed SMA, h-caldesmon, and collagen IV. Malformed vascular endothelial cells expressed CD31, CD34 and ERG. No postoperative recurrence was found in the 12 cases. Conclusions: GVM is an uncommon type of simple venous malformation in the superficial soft tissue and different from the classical glomus tumor. Morphologically, one or more layers of glomus cells grow around the dilated venous malformation-like lumen, which can be combined with common venous malformations.


Assuntos
Masculino , Feminino , Humanos , Criança , Tumor Glômico/cirurgia , Células Endoteliais/patologia , Paraganglioma Extrassuprarrenal/patologia , Imuno-Histoquímica
2.
International Journal of Biomedical Engineering ; (6): 73-79,85, 2022.
Artigo em Chinês | WPRIM | ID: wpr-954195

RESUMO

Theranotics, an integrated diagnosis and treatment nanoplatform technology based on nanomaterials, integrates the diagnosis and treatment of diseases seamlessly and shows a broad prospect in medical practice. With the rapid development of nanomedicine, the technology of magnetic nanoparticles (MNPs) synthesis is becoming more and more mature. MNPs have controllable shape and particle size, admirable stability and biocompatibility, excellent magnetic properties, and can be readily chemically modified. These advantages make them widely used in clinical practice, such as diagnosis, targeted drug delivery, medical imaging, hyperthermia therapy, and radiotherapy, which also make MNPs high-quality materials for integrated diagnosis and treatment platforms.In this paper, the research progress of MNPs in the areas of magnetically guided drug delivery, magnetic thermotherapy, and multimodal imaging was reviewed; their advantages as an integrated platform for diagnosis and treatment were discussed; and the problems faced in research and application prospects were summarized and outlooked.

3.
Chinese Journal of Pathology ; (12): 202-206, 2022.
Artigo em Chinês | WPRIM | ID: wpr-935505

RESUMO

Objective: To investigate the clinical and pathologic features, diagnosis and differential diagnosis of congenital hemangioma (CH). Methods: Forty cases of CH were diagnosed from January 2017 to December 2020 in Henan Provincial People's Hospital. The clinical and pathological and immunohistochemical data were analyzed, with review of literature. Results: There were 24 male and 16 female patients. The lesions were located in the head, neck (11 cases), limbs (14 cases), and trunk (15 cases). The clinical manifestations were congenital painless plaques or masses, the larger ones protruded on the skin surface, mostly dusky purple or bright red, with surrounding white halos. Under low magnification, the tumor was lobular and well demarcated, composed of neo-microvascular lumen of different sizes. The vascular endothelial cells were cuboidal or hobnail in appearance, forming stellar drainage vessels within the lobules. Extra-medullary hematopoiesis was seen in one case of rapidly involuting CH; there were different number of tortuous and dilated vascular lumen between the lobular structures, and some non-involuting CH cases were vascular malformations, which were devoid of lobulated structures. Immunohistochemistry showed that endothelial cells were strongly positive for CD31, CD34 and ERG, while D2-40 and GLUT-1 were negative. Conclusions: CH is a benign congenital vascular tumor with characteristic lobulated growth and abnormal blood vessels in the stroma. Pathological diagnosis often needs to be differentiated from infantile hemangioma, pyogenic granuloma, kaposiform hemangioendothelioma and vascular malformation.


Assuntos
Feminino , Humanos , Masculino , Células Endoteliais/patologia , Hemangioendotelioma/patologia , Hemangioma/patologia , Síndrome de Kasabach-Merritt/patologia , Sarcoma de Kaposi/patologia , Neoplasias Cutâneas/patologia
4.
Chinese Health Economics ; (12): 78-80, 2018.
Artigo em Chinês | WPRIM | ID: wpr-703488

RESUMO

According to The Budget Law,the hospital needed to compile the government department budget.According to The Hospital Financial System,the hospital needed to compile a comprehensive budget.Because of the institutional reasons,there were existing problems,such as the unclear limit of government budget constraint on hospital,accountants did not understand the government department budget and the comprehensive budget,the budget system and the accounting system were not coordinated.Suggestions were prooosed to solve the problems.

5.
Chinese Journal of Clinical and Experimental Pathology ; (12): 1356-1359, 2017.
Artigo em Chinês | WPRIM | ID: wpr-695051

RESUMO

Purpose To study the infection mode and distribution of high risk human papillomavirus (HR-HPV) in lowgrade squamous intraepithelial lesions (LSILs) and to analysis its effect.Methods HPV genotype of 328 cervical tissues embedded by paraffin which contained 168 LSILs and 160 highgrade squamous intraepithelial lesions (HSILs) were detected by the methods of real-time fluorescence quantitative PCR and in situ hybridization.Results Diseased cells of LSIL stained with hematoxylin and eosin(HE) were concentrated in the upper layer of the cervical squamous epithelium,whereas HSIL were concentrated in the lower layer of the epithelium in HE staining.In LSIL,the positive ratcs of HR-HPV was 95.2%.In LSIL and HSIL,HPV 16/18 positive rates were 26.2%,57.5%,respectively.Other HR-HPV positive rates were 80.9%,55.0%,respectively.The differences were statistically significant (P <0.001).HR-HPV positive rate was 70.2% in the group of in situ hybridization.The positive cells were confined to the surface and middle layer of the LSIL cervical squamous epithelium.Conclusion HR-HPV infection is closely related to LSIL,especially other HR-HPV (except for HPV 16/18).The infection mode of HR-HPV in LSIL starts from the surface layer of the cervical squamous epithelium.

6.
China Journal of Chinese Materia Medica ; (24): 1213-1219, 2017.
Artigo em Chinês | WPRIM | ID: wpr-350201

RESUMO

This article reviewed the current situation of ethnomedicines in China, and propos that the important research on ethnomedicine is to focus on the theoretical and empirical characteristics of ethnomedicine and the special features in the processes of collecting,processing and clinical application of ethnomedicine, which will be benefit to find the research principles and methods with the characteristics of ethnomedicine. These principles include making clear of the effective substances and mechanisms, and theoretical and empirical characteristics of ethnomedicine. The three methods in the new drug discovery technology system for ethnomedicine are to solve the difference in varieties of traditional medical systems depending on finding the similarities of them, to find the conjunction in varieties of traditional medical systems through the key-herbs, and to find medicinal sources in varieties of traditional medical systems according to pharmaphylogeny.

7.
Chinese Journal of Current Advances in General Surgery ; (4): 505-510, 2017.
Artigo em Chinês | WPRIM | ID: wpr-660406

RESUMO

Objective:To investigate the inhibitory effect of Shenqi Fuzheng injection on the expression of VEGF gene in human hepatocellular carcinoma cell line MHCC97L,and to elucidate the effect of Shenqi Fuzheng injection on tumor invasiveness,and to validate it through siRNA technology.Methods:After 24 hours of addition of Shenqi Fuzheng Injection (MHC) in the culture system of MHCC97L,Real-time PCR and Western-blot were used to detect the expression of MMP-9 and MMP-9 in the control group (Shenqi Fuzheng),negative control group (NaCI) And the expression of VEGF and RNA in the blank control group,and to determine the effect of Shenqi Fuzheng injection on the expression of VEGF and RNA.The effect of Shenqi Fuzheng Injection on the invasiveness of MHCC97L cells was detected by Transwell assay.The lentiviral vectors of MHCC97L cells were transfected into 293T cells and transfected into MHCC97L cells.The transfected cells were transfected into 293T cells by Lipofectamine 2000,and transfected into MHCC97L cells.The Effect of Shenqi Fuzheng Injection on the Invasion of VEGFi Tumor by Transwell.Results:Shenqi Fuzheng injection could significantly decrease the expression of VEGF mRNA and protein in normal MHCC97L cells (P<0.05),and decrease the invasiveness of tumor cells (P<0.05).VEGF RNA interference was successful.After intervention,Shenqi Fuzheng injection could not decrease the invasiveness of MHCC97L tumor (P>0.05).Conclusion:Shenqi Fuzheng injection can reduce the invasion of MHCC97L cells by inhibiting the expression of VEGF gene.

8.
Chinese Journal of Current Advances in General Surgery ; (4): 505-510, 2017.
Artigo em Chinês | WPRIM | ID: wpr-657890

RESUMO

Objective:To investigate the inhibitory effect of Shenqi Fuzheng injection on the expression of VEGF gene in human hepatocellular carcinoma cell line MHCC97L,and to elucidate the effect of Shenqi Fuzheng injection on tumor invasiveness,and to validate it through siRNA technology.Methods:After 24 hours of addition of Shenqi Fuzheng Injection (MHC) in the culture system of MHCC97L,Real-time PCR and Western-blot were used to detect the expression of MMP-9 and MMP-9 in the control group (Shenqi Fuzheng),negative control group (NaCI) And the expression of VEGF and RNA in the blank control group,and to determine the effect of Shenqi Fuzheng injection on the expression of VEGF and RNA.The effect of Shenqi Fuzheng Injection on the invasiveness of MHCC97L cells was detected by Transwell assay.The lentiviral vectors of MHCC97L cells were transfected into 293T cells and transfected into MHCC97L cells.The transfected cells were transfected into 293T cells by Lipofectamine 2000,and transfected into MHCC97L cells.The Effect of Shenqi Fuzheng Injection on the Invasion of VEGFi Tumor by Transwell.Results:Shenqi Fuzheng injection could significantly decrease the expression of VEGF mRNA and protein in normal MHCC97L cells (P<0.05),and decrease the invasiveness of tumor cells (P<0.05).VEGF RNA interference was successful.After intervention,Shenqi Fuzheng injection could not decrease the invasiveness of MHCC97L tumor (P>0.05).Conclusion:Shenqi Fuzheng injection can reduce the invasion of MHCC97L cells by inhibiting the expression of VEGF gene.

9.
China Journal of Chinese Materia Medica ; (24): 1759-1761, 2015.
Artigo em Chinês | WPRIM | ID: wpr-351269

RESUMO

Endophytic fungi Penicillium dangeardii, isolated from Lysidice rhodostegia Hance root, was fermented and the secondary metabolites were studied. By means of Sephadex LH-20 column chromatography, ODS column chromatography and PHPLC over the fermented culture, 5 compounds were isolated. By using ESI-MS and NMR, the structures of the compounds were determined as N-[9-(β- D-ribofuranosyl)-9H-purin-6-yl]-L-aspartic acid (1), 3-caffeoylquinic acid (2), 4-caffeoylquinic acid (3), and 5-caffeoylquinic acid (4), 3-hydroxy-benzoic acid-4-O-β-D-glucopyranoside (5).


Assuntos
Fatores Biológicos , Química , Metabolismo , Endófitos , Química , Metabolismo , Fabaceae , Microbiologia , Fermentação , Estrutura Molecular , Penicillium , Química , Metabolismo , Metabolismo Secundário
10.
China Journal of Chinese Materia Medica ; (24): 1893-1897, 2015.
Artigo em Chinês | WPRIM | ID: wpr-351244

RESUMO

Valeriana jatamansi (syn. V. wallichii), a traditional Chinese medicine recorded in Chinese Pharmacopeia (1977 and 2010 edition), has been used for treatment of a variety of conditions including sleep problems, obesity, nervous disorders, epilepsy, insanity, snake poisoning, eye trouble, and skin diseases. Also, it was used as an important substitute for the European V. officinalis, whose root preparation, popularly known as valerian, has been employed as a mild sedative for a long time. In recent years, much attention has been draw to the iridoids, one of the major bioactive constituents of V. jatamansi, leading to the discovery of a series of new iridoids with anti-tumor and neuroprotective activities. Their action machnism also has been discussed. This paper summerized the iridoids and their bioactivities from V. jatamansi in recent years, which could provide basic foundation for development and research of V. jatamansi.


Assuntos
Animais , Humanos , Medicamentos de Ervas Chinesas , Química , Farmacologia , Iridoides , Química , Farmacologia , Valeriana , Química
11.
Chinese Journal of Urology ; (12): 721-725, 2015.
Artigo em Chinês | WPRIM | ID: wpr-479750

RESUMO

Objective To evaluate the feasibility and outcomes of robotic-assisted laparoscopic pyeloplasty in children .Methods A retrospective study was performed in patients who underwent robotic-assisted laparoscopic pyeloplasty ( Anderson-Hynes ) at our institution between January 2014 to August 2014.Totally 6 boys were diagnosed as left ureteropelvic junction obstruction depending on the symptoms and radiographic studies .The mean age was 9 years ( range 4 -12 years ) .Results The procedure was performed successfully without conversion to open surgery in all of the cases .Mean operative time was 216 min (range 175-269 min), with a mean robotic anastomosis time of 45 min (range 30-60 min).Mean estimated blood loss was less than 15 ml.The mean hospitalization was 4.5 days.Mean follow-up period was 10 months ( range 7 -14 months ) .There were no perioperative complications , and recovery was uncomplicated (without recurrence, pyelonephritis, nephrarctia) in all of the patients.Conclusion Robotic-assisted laparoscopic pyeloplasty can be safely performed in children older than 4-year-old with ureteropelvic junction obstruction .

12.
China Journal of Chinese Materia Medica ; (24): 3974-3977, 2014.
Artigo em Chinês | WPRIM | ID: wpr-310955

RESUMO

The PDB culture medium was selected to ferment the endophyte strain, and the secondary metabolites of endophytic fungi Penicillium polonicum were studied. Combined application of Sephadex LH-20, ODS and HPLC chromatographies over the ethyl acetate extract of the fermented culture led to the isolation of 6 compounds. By spectral methods, the structures were elucidated as [3, 5-dihydroxy-2-(7-hydroxy-octanoyl)]-ethylphenylacetate (1), (3, 5-dihydroxy-2- octanoyl)-ethyl phenylacetate (2), (5, 7-di- hydroxy-9-heptyl)-isobenzo pyran-3-one (3), 3-(hydroxymethyl) 4-(1E)-1- propen-1-yl-(1R, 2S, 5R, 6S)-7-oxabicyclo [4.1.0] hept-3-ene-2, 5-diol (4), (E)-2-methoxy-3-(prop-1-enyl) phenol (5) and p-hydroxylphenylethanol (6).


Assuntos
Fatores Biológicos , Química , Metabolismo , Endófitos , Química , Metabolismo , Fabaceae , Microbiologia , Fermentação , Penicillium , Química , Metabolismo , Metabolismo Secundário
13.
China Journal of Orthopaedics and Traumatology ; (12): 331-334, 2012.
Artigo em Chinês | WPRIM | ID: wpr-321908

RESUMO

<p><b>OBJECTIVE</b>To explore the curative effect of intramedullary nailing pressure within a fixed period with a bone graft taken from the opening of intramedullary nailing on tibial fracture nonunion.</p><p><b>METHODS</b>From February 2008 to October 2010, 18 patients with nonunion of tibial shaft were treated by pressurized intramedullary nail fixation and bone grafting taken from the opening of intramedullary nailing. They included 12 males and 6 females ranging in age from 31 to 67 (mean 42) years. The time from injury to surgery was 6-18 months (mean 8 months). There were 11 cases of nonunion and 7 cases of delayed union. After operation, the knee function was evaluated according to HSS scoring. The therapeutic effect was assessed by Tenny-Wiss scoring system.</p><p><b>RESULTS</b>All patients were followed up for 12-36 (mean 18) months. The wound healed well without infection or skin necrosis. All patients had no nonunion, infection, deformity, and re-fracture. The bone healing time was from 4 to 8 months (mean 6 months). The HHS score of knee function 1 year after surgery averaged (89.97 +/- 3.21). The effect result of Tenny and Wiss scoring system was excellent in 16 cases and good in 2 cases.</p><p><b>CONCLUSION</b>Open reduction by intramedullary nail fixed within a period with bone graft (from the opening of intramedullary nailing) for treatment of nonunion and delayed union of tibial fracture can improve the fracture healing rate, avoid complications from iliac bone grafting, and reduce medical expenses.</p>


Assuntos
Adulto , Idoso , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Transplante Ósseo , Fixação Intramedular de Fraturas , Métodos , Consolidação da Fratura , Fraturas não Consolidadas , Cirurgia Geral , Fraturas da Tíbia , Cirurgia Geral
14.
China Journal of Chinese Materia Medica ; (24): 3083-3085, 2012.
Artigo em Chinês | WPRIM | ID: wpr-337988

RESUMO

Aspergillus fumigatus, a type of endophytic fungi from Erthrophleum fordii, was fermented with GPY culture medium. Fermented liquid and mycelium were extracted from fermented products after freezing and thawing treatment. After alcohol extraction, mycelium was extracted with ethyl acetate and n-butyl alcohol, respectively. According to the results of cytotoxity of tumor cells, ethyl acetate extracts were studied for their chemical constituents. Five diketopiperazine compounds were separated and purified with silica gel, MCI and Sephadex LH-20 column chromatography, reversed-phase chromatographic column and preparative HPLC, their structures were identified as cyclo- (R-Pro-R-Phe) (1), cyclo- (trans-4-OH-D-Pro-D-Phe) (2), cyclo- (R-Tyr-S-Ile) (3), cyclo-(R-Phe-S-Ile) (4), and cyclo-(R-Val-S-Tyr) (5) by using spectral methods.


Assuntos
Humanos , Aspergillus fumigatus , Química , Metabolismo , Linhagem Celular Tumoral , Dicetopiperazinas , Química , Metabolismo , Farmacologia , Endófitos , Química , Metabolismo , Fabaceae , Microbiologia , Micélio , Química , Metabolismo
15.
Microbiology ; (12)2008.
Artigo em Chinês | WPRIM | ID: wpr-685984

RESUMO

In order to increase the production of insecticidal crystal proteins Cry1 and Cry2, firstly, Plack-ett-Burman design was applied to evaluate the effectiveness of the related nutrition factors; it was found that the soybean powder and MnSO4?H2O were significant factors for Cry1 production, but the yield of Cry2 wasn’t effected remarkably in such medium. Then the steepest ascent experiment was adopted to approach the optimal region of the medium composition. Lastly, the optimal concentration of the soybean powder and MnSO4?H2O was 11.5 and 0.02 g/L, obtained by response surface methodology (RSM). The final yields of Cry1 and Cry2 was 0.32 mg/mL and 0.11 mg/mL, increasing twice more than that in the medium optimized before. The median lethal concentration (LC50) of optimal medium was 1.09 ?L/mL. The toxicity to Heli-coverpa armigera was significantly enhanced than the old one.

16.
Chinese Journal of Plastic Surgery ; (6): 109-111, 2007.
Artigo em Chinês | WPRIM | ID: wpr-297085

RESUMO

<p><b>OBJECTIVE</b>To investigate the feasibility and results of application of both expanded cutaneous flap and temporoparietal fascia flap in total ear reconstruction with Medpor framework.</p><p><b>METHODS</b>The main procedure consists of two stages: Stage I-skin expansion; Stage II -auricle formation consists of orientation of Medpor implant and creation of coverage for the implant by both expanded skin flap and temporoparietal fascia flap.</p><p><b>RESULTS</b>Twenty-two ears in 22 unilateral microtia patients were constructed using Medpor implants covered with both expanded cutaneous flap and temporoparietal fascia flap over the last three years, they were accepted as pleasing by the patients.</p><p><b>CONCLUSIONS</b>Application of both expanded cutaneous flap and temporoparietal fascia flap can assure no extrusion of Medpor implant in ear reconstruction. Either more, the two layers of transferred tissues will not affect the profile details of the reconstructed ear. And because the skin covering the framework and fascia is derived from mastoid region, the appearance and profile of the reconstructed auricle is true to nature and close to that of the opposite one.</p>


Assuntos
Adolescente , Adulto , Criança , Feminino , Humanos , Masculino , Adulto Jovem , Materiais Biocompatíveis , Orelha Externa , Cirurgia Geral , Fáscia , Transplante , Polietilenos , Implantação de Prótese , Procedimentos de Cirurgia Plástica , Métodos , Transplante de Pele , Stents , Retalhos Cirúrgicos , Osso Temporal
17.
China Journal of Chinese Materia Medica ; (24): 224-226, 2007.
Artigo em Chinês | WPRIM | ID: wpr-246004

RESUMO

<p><b>OBJECTIVE</b>To establish a HPLC method for bufothionine in the skin of Bufo bufo gargarizans and Huachansu injection.</p><p><b>METHOD</b>The samples were separated using a Lichrosob C18 column with CH3CN-H2O (10:90) as mobile phase. Flow rate was at 1.0 mL x min (-1) and the detection wavelength was at 225 nm.</p><p><b>RESULT</b>The calibration curve of bufothionine was linear over the range of 0.0772-0.4632 microg and the average recovery was 99. 2%. The contents of bufothionine were fluctuated from 36.4-641.8 microg x g(-1) in the skin of Bufo bufo gargarizans and 22.47-33.16 microg x mL(-1) in Huachansu injection, respectively.</p><p><b>CONCLUSION</b>The contents of bufothionine were greatly different between cultured and wild species. The method was suitable for the quality control of the skin of Bufo bufo gargarizans and its preparation.</p>


Assuntos
Animais , Bufo bufo , China , Cromatografia Líquida de Alta Pressão , Métodos , Injeções , Materia Medica , Química , Fenotiazinas , Controle de Qualidade , Reprodutibilidade dos Testes , Pele , Química
18.
China Journal of Chinese Materia Medica ; (24): 289-290, 2006.
Artigo em Chinês | WPRIM | ID: wpr-350958

RESUMO

<p><b>OBJECTIVE</b>To establish a method of the quantitative determination of acetylharpagide in Ajuga decumbens.</p><p><b>METHOD</b>The chromatographic conditions were as follows: a Phenomenex Luna C18 column was used, the mobile phase was composed of acetontrile-water (15:85), the flow rate was 1.0 mL x min(-1) and the UV absorbance detection was set at 197 nm.</p><p><b>RESULT</b>Linearity of acetylharpagide was in the range of 0.6-3.6 microg (r = 0.9993), and the average recovery and RSD were 99.13% and 2.48%, respectively.</p><p><b>CONCLUSION</b>The contents of acetylharpagide ranged 0.40%-6.39% in A. decumbens. The method was simple, accurate and sensitive.</p>


Assuntos
Ajuga , Química , Cromatografia Líquida de Alta Pressão , Métodos , Medicamentos de Ervas Chinesas , Plantas Medicinais , Química , Piranos , Reprodutibilidade dos Testes , Sensibilidade e Especificidade
19.
China Journal of Chinese Materia Medica ; (24): 408-410, 2004.
Artigo em Chinês | WPRIM | ID: wpr-256343

RESUMO

<p><b>OBJECTIVE</b>To develop a HPLC method to determine the contents of aristolochic A in aristolochia debilis and Asarun spp..</p><p><b>METHOD</b>Methanol-water-formic acid extracts were separated on an Alltech C18 column with methanol-water-acetic acid (68:32:1) as mobile phase. The flow rate was 1.0 mL x min(-1). UV detection wavelength was 390 nm. Column temperature was 35 degrees C.</p><p><b>RESULT</b>Aristolochic acid A was separated well. The relationship of injection amounts and peak areas was linear (r = 0.9999) the range of 0.12-1.89 microg x g(-1) and the recovery rate was 101.8% (n = 5). 11 samples of aristolochia debilis which bought from different areas in China were determined, and the contents of aristolochic acid A varied from 0.9 to 2 mg x g(-1). The difference of the contents in Asarum spp. was obvious. The highest is 0.35, and aristolochic acid A couldn't be detected in one sample.</p>


Assuntos
Aristolochia , Química , Ácidos Aristolóquicos , Asarum , Química , China , Cromatografia Líquida de Alta Pressão , Métodos , Ecossistema , Raízes de Plantas , Química , Plantas Medicinais , Química
20.
Chinese Journal of Biotechnology ; (12): 656-661, 2004.
Artigo em Chinês | WPRIM | ID: wpr-249959

RESUMO

The CrylA Crystal Protein from Bacillus thuringiensis is associated with DNA, but the role and sequences of these DNA molecules are unknown. CrylA bipyramidal crystals from B. thuringiensis strain 4.0718 was selectively dissolved and associated DNA was extracted from protoxin. The DNA was digested with Nde I to obtain 3 to 5 kb fragments and then the fragments were subcloned into pMD18-T vector, screening of recombinants were done by PCR-RFLP and sequencing. The ORF of cry1Ac gene was amplified by primers designed and then subcloned. The 3.5 kb BamH I and Sal I fragments of pMDX35 was inserted into the pET30a vector, giving 8.9 kb recombinant plasmid, pETX35. ETX35 strain were obtained by transformed pETX35 into B121 (DE3). A 141 kD fusion protein was superexpressed as inclusion bodies. Quantitative protein analysis indicated that the amount of 141 kD protein was above the level of 51.36% of total cellular protein. Plasmid pHTX42 constructed from shuttle vector pHT304 was transformed B. thuringiensis acrystalliferous strain XBU001 with electroporation to obtain the recombinant HTX42. The recombinant protein was found with a molecular mass of 130 kD on sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Scanning analysis indicated that the expressed protein accounted up to 79.28% of total cellular proteins and accumulated in the cells mounted up to 64.13% of cellular dry weight. Under Atomic Force Microscopy (AFM), typical bipyramidal crystals from HTX42 strain were found with a size of 1.2 microm x 2.0 microm. Bioassay showed that these inclusion bodies of ETX35 strain and crystals from HTX42 strain were highly toxic against the larvae of Plutella xylostella. On such a base, constructing insecticidal recombinant and analyzing the source, structure, and function of the 20 kb DNA can be further achieved.


Assuntos
Animais , Bacillus thuringiensis , Genética , Proteínas de Bactérias , Genética , Farmacologia , Clonagem Molecular , Endotoxinas , Genética , Farmacologia , Proteínas Hemolisinas , Genética , Farmacologia , Microscopia de Força Atômica , Mariposas , Plasmídeos , Proteínas Recombinantes , Farmacologia
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