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Objective: To investigate the efficacy and safety of pedunculated rectus abdominis combined with bilateral ureteral extravestheter drainage in the treatment of refractory bladder-vaginal stump fistula. Methods: The clinical data of 8 cases of the refractory bladder-vaginal stump fistula were admitted to the Second Hospital of Hebei Medical University and Henan Cancer Hospital and underwent the clinical treatment of bladder-vaginal stump from December 2019 to December 2022 were collected. The reason of refractory bladder-vaginal stump fistula was analyzed, the operation manner of pedunculated rectus abdominis combined with peduncle and bilateral ureter for the treatment of bladder-vaginal stump through extrabladder drainage was explored. The operation time, bleeding volume and clinical effect were record. Results: The median operation time of 8 patients was 150 minutes(120~180 min), and the median blood loss was 400 ml(200~600 ml). During the perioperative period, there were 2 cases of incision infection, delayed healing by debridement and dressing, 2 cases of incision rupture and suture wound healing after reoperation, and 2 cases of urinary tract infection were cured by anti-infection. When followed up for 6 months, 8 cases of vesicovaginal stump fistula were cured. Conclusion: Bilateral ureteral external drainage of the rectus abdominis muscle, has a practical effect in the treatment of refractory bladder-vaginal stump fistula, which can be one of the clinical repairing treatment.
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Femenino , Humanos , Vejiga Urinaria/cirugía , Uréter/cirugía , Recto del Abdomen , Drenaje , FístulaRESUMEN
Objective: To investigate the efficacy and safety of pedunculated rectus abdominis combined with bilateral ureteral extravestheter drainage in the treatment of refractory bladder-vaginal stump fistula. Methods: The clinical data of 8 cases of the refractory bladder-vaginal stump fistula were admitted to the Second Hospital of Hebei Medical University and Henan Cancer Hospital and underwent the clinical treatment of bladder-vaginal stump from December 2019 to December 2022 were collected. The reason of refractory bladder-vaginal stump fistula was analyzed, the operation manner of pedunculated rectus abdominis combined with peduncle and bilateral ureter for the treatment of bladder-vaginal stump through extrabladder drainage was explored. The operation time, bleeding volume and clinical effect were record. Results: The median operation time of 8 patients was 150 minutes(120~180 min), and the median blood loss was 400 ml(200~600 ml). During the perioperative period, there were 2 cases of incision infection, delayed healing by debridement and dressing, 2 cases of incision rupture and suture wound healing after reoperation, and 2 cases of urinary tract infection were cured by anti-infection. When followed up for 6 months, 8 cases of vesicovaginal stump fistula were cured. Conclusion: Bilateral ureteral external drainage of the rectus abdominis muscle, has a practical effect in the treatment of refractory bladder-vaginal stump fistula, which can be one of the clinical repairing treatment.
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Femenino , Humanos , Vejiga Urinaria/cirugía , Uréter/cirugía , Recto del Abdomen , Drenaje , FístulaRESUMEN
Globally, more than 2 million bone grafts are performed every year for bone defects in orthopedics, neurosurgery, and dental procedures. Current treatment options include the use of grafts of human, animal or synthetic origin. In this case, autograft is the current gold standard. However, its quantity is limited, a second wound(donor site) needs to be created, and the risk of infection, pain, and morbidity increases. In recent years, the rise of tissue engineering and 3D bioprinting has provided a new idea for treating bone defects in patients. 3D bioprinting is a branch of the applications of "additive manufacturing" in biological tissue engineering. It can precisely control cells, personalize macro and micro structures as needed, and can be used in bone regeneration applications. The establishment of osteoblast scaffolds is the basis of 3D bioprinting, and hydrogels suitable for the growth of bone and cartilage are the basis of scaffold research. For this reason, domestic and foreign scholars have developed and researcheda variety of hydrogel scaffolds, and they have found that mixed hydrogels with multiple biological materials have more advantages than single-material hydrogels. For example, hydroxyapatite, alginate or hyaluronic acid is used as the main component to mix several or more bioprinting materials, and 3D printed bone scaffold formed after combining the required cells can promote bone growth and differentiation better than traditional scaffolds. As the printed structure becomes thicker, the diffusion of nutrients and oxygen becomes more and more difficult. This is especially true in the reconstruction of bone tissue and it is necessary to create an interconnected and effective vascular network. Therefore, the formation of blood vessels in the stent is indispensable. This article mainly reviews the step-by-step research progress of bone printing scaffold materials and vascular network formation in 3D bioprinting.
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Objective To explore the clinical characteristics ,diagnosis ,treatment and prognosis of parainfluenza virus (PIV) pneumonia after lung transplantation .Methods One case of PIV pneumonia after lung transplantation was retrospectively analyzed . The relevant domestic and foreign cases and literature review were summarized .Results The recipient underwent sequential bilateral lung transplantation for chronic obstructive pulmonary disease ,bullae and respiratory failure .Donor lung was sourced from donation after cardiac death .Routine anti-rejection therapy was prescribed postoperatively .At 14 months ,cough and shortness of breath lead to hospitalization for over 1 month .At 15 months ,sputum/fungal smear and culture showed that nucleic acid of PIV was positive . The definite diagnosis was PIV pneumonia after lung transplantation .After ribavirin antiviral therapy ,tracheal intubation and invasive ventilation ,followed by imipenem plus doxycycline plus anti-infective therapy ,ganciclovir antiviral therapy ,repeated bronchoscopic sputum aspiration and lavage treatment ,the patient's condition deteriorated and died from breathing failure and septic shock at 16 months .Conclusions Preventing PIV infection after lung transplantation is of vital importance .PCR is essential for a rapid detection of virus infection .However ,there is no curative treatment of PIV infection .Specific parainfluenza immunoglobulin and DAS 181 aerosol inhalation may be applied for future treatment of PIV infection in lung transplant recipients .
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Objective: To screen the active fraction of Euscaphis japonica roots with inhibitory activity on hepatic lipid accumulation and investigate its chemical constituents. Methods: Different polar fractions were prepared by extraction with organic solvents, oil Red O staining and triglyceride content assay were used to determine inhibitory activity of different polar fraction on oleic acid induced triglyceride accumulation on HepG2 cells, and the constituents of active fraction were isolated and purified by various chromatography techniques such as column chromatography on silica gel, Sephadex LH-20, and HPLC, and their structures were identified by physicochemical properties and spectral data. Results: The petroleum ether fraction exhibited significantly inhibitory activity on oleic acid induced triglyceride accumulation on HepG2 cells. Nine compounds were isolated and identified as 3β, 19- dihydroxy-24-trans-ferulyloxyurs-12-en-28-oic acid (1), β-sitosterol (2), 7-hydroxy-2-octen-5-olide (3), 3, 3'-dimethoxy-ellagic acid (4), vanillin (5), ethyl-5-oxo-tetrahydro-3-furancarboxylate (6), gallic acid (7), 3, 3'-di-O-methylellagic acid 4-(5″-acetyl)-α-L- arabinofuranoside (8), and bergapten (9). Conclusion: The petroleum ether fraction is main active fraction. The compounds 1, 6, 8, and 9 are obtained from genus Euscaphis Sieb. et Zucc. for the first time.
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Objective To investigate the effects and cost of four formulations of niclosamide ethanolamine salt in Oncomela-nia hupensis snail control in the field in marshland and lake regions,so as to provide the evidence for drawing up the plan of mol-luscicide using in schistosomiasis endemic areas. Methods One drainage channel and one channel without water in the same area with snails in Jiangling County,Jingzhou City were selected as the research fields. The drainage channel was divided into 9 sections,except one section as a blank control group where the natural death rate of snails was observed only,and the remaining 8 sections were taken as the observation groups,where different dosages of 4%niclosamide ethanolamine salt powder,5%ni-closamide ethanolamine salt granules,25% niclosamide ethanolamine salt suspending agent,26% metaldehyde and ni-closamide ethanolamine salt suspending agent,and 50%niclosamide ethanolamine salt wettable powder were used re-spectively. The channel without water were divided into 4 sections,except one section as a blank control group,the oth-er 3 segments were taken as the observation groups,where 4%niclosamide ethanolamine salt powder,5%niclosamide ethanolamine salt granules,and 50% niclosamide ethanolamine salt wettable powder were used respectively. Before and after spraying molluscicide for 7 days and 15 days,the system sampling method was used to observe the effects of snail control. Meanwhile,the unit cost method was used to calculate the costs of the different mulluscicide formulations abovementioned in unit area(1 m2). Results In the field at the drainage channel,the snail mortality rates of the groups spraying 4%niclosamide ethanolamine salt powder(50 g/m2),5%niclosamide ethanolamine salt granules(40 g/m2),25% niclosamide ethanolamine suspending agent,26% metaldehyde and niclosamide ethanolamine salt sus-pending agent,and 50%niclosamide ethanolamine salt wettable powder(2 g/m2 and 4 g/m2)for 7 days were 79.52%-97.87%,while the rates after spraying for 15 days were 71.00%-96.30%,and compared with those before spraying, the differences were statistically significant(all P<0.01). For the groups spraying with 2 g/m2 or 4 g/m2 suspending agent as well as wettable powder for 7 days,the snail mortality rates were significantly different(both P<0.05). In the field at the channel without water,the snail mortality rates of the 3 observation groups after spraying molluscicide for 7 days were 97.14%-100%,while for 15 days were 94.32%-100%,and compared with the rates before spraying,all the differences were statistically significant(all P<0.01). The unit costs per 1 m2 of the molluscicide abovementioned were ranged from 0.280 Yuan to 0.416 Yuan. Conclusions In marshland area inside embankment,the molluscicide formulations of the powder and granule are suitable for the environments without water or with instability water level , while the molluscicide formulations of the suspended agents and wettable powder are suitable for the water environment. Though the unit cost of powder is the lowest,the molluscicide in this formulation flies away seriously.
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Objective To evaluate the efifcacy and safety of hemostatics of injected gelatin matrix (HIGM) under the guidance of contrast-enhanced ultrasound (CEUS) for treating splenic trauma in canine model. Methods A total of 24 commercial hybrid dogs underwent celiotomy with creation of uniformly blunt splenic trauma lesion of 4.0 cm×4.0 cm×2.5 cm (length, width and depth, respectively) by hemostatic clamp. Subjects were prospectively randomized into two groups. The treatment group was treated with HIGM under the guidance of CEUS and the positive control group received thrombin solution. Conventional ultrasound and CEUS were performed to record the ascites and the splenic lesion areas at 1st, 3rd, 7th, 14th and 21st day. The ifne needle biopsy and splenectomy were performed for histopathologic examination. The weight, free intraperitoneal lfuid and injury site were compared with t test between HIGM and postive group. Results All animals in two groups survived. All dogs stopped hemorrhage after injection of HIGM under CEUS guidance. The area of injury site was (12.91±0.89) cm2, (4.45±0.75) cm2 and (1.38±0.23) cm2 at 1st, 3rd and 7th day and splenic lesions were not found at 14th and 21st day in all dogs (n=12) of HIGM group. The splenic lesion was (16.74±0.91) cm2, (11.26±0.99) cm2, (8.02±0.82) cm2 and (1.58±0.36) cm2 in the postive group at 1st, 3rd, 7th and 14th day and splenic lesions were not found at 21st day in all dogs (n=12). At 7th and 14th day post-injection, lesion areas were statistically significant between two groups (t=27.162, P=0.008;t=15.129, P=0.001). Free intraperitoneal lfuid was (0.91±0.05) cm at 1st day detected by conventional ultrasound and free intraperitoneal fluid was not found at 3rd, 7th, 14th and 21st day in all dogs (n=12) of HIGM group. The free intraperitoneal fluid in thepositive group was (1.96±0.17) cm, (1.30±0.11) cm and (0.81±0.12) cm at 1st, 3rd and 7th day and free intraperitoneal lfuid was not found at 14th and 21st day in all dogs (n=12). At 1st, 3rd and 7th day post-injection, free intraperatitoneal lfuid was statistically significant between two groups (t=20.934, P=0.003; t=41.310, P=0.000; t=22.520, P=0.000). Histopathological examination showed that there was no foreign body and foreign body granuloma and the structure of red pulp was recovered at 7th, 14th and 21st day. Gross anatomy showed that the splenic injury site was recovered completely without complications. Conclusion This study explored the value of HIGM for splenic trauma and provided a preliminary experimental evidence for clinical treatment.
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Objective To analyze the clinical characteristic of autoimmune gastritis (AIG).Methods From January 1990 to April 2010,the clinical data of 55 AIG patients were retrospectively analyzed,which included hemoglobin,lactate dehydrogenase (LDH),α-hydroxybutyrate dehydrogenase (α-HBDH),gastrin,intrinsic factor antibody (IFA),parietal cell antibody (PCA),gastrointestinal endoscopy examination and 24-hour esophageal pH recording.Another 31 megaloblastic anemia (MA) patients were selected as control.Statistical analysis was performed by independent-samples t test.Results Among 55 AIG patients,49 patients were associated with MA,and three out of four cases were identified of IFA.About 43.8% (21/48) patients were PCA positive.Before treatment,the levels of LDH and α-HBDH of AIG patients with MA were (1045.50±853.46)U/L and (853.71±824.23) U/L which significantly increased,than those of patients without MA [(166.67±41.03) U/L,(133.67±27.90) U/L],the differences were statistically significant (t=-4.665,-2.120,both P<0.05),however there was no significant difference when compared with the control group [(1047.52±1028.31) U/L,(1050.23±1264.37) U/L,both P>0.05)].A total of 46 patients underwent gastroendoscopy examination,63.0% (29/46) patients had gastric body atrophy while gastric antrum not involved; 34.8% (16/46) patients had neither gastric body nor antrum atrophy; seven patients gastric mucosal showed intestinal metaplasia and one patient showed intestinal metaplasia with atypical hyperplasia and 2.2% (1/46) presented both the antrum and the body atrophy.Conclusions The levels of LDH and α-HBDH increased in AIG patients might be related with MA caused marrow in-situ hemolysis.IFA is recommended as a routine test for AIG.There is still some limitations of AIG diagnosis according to histopathological features of gastric endoscopy specimen.The clinical features should be taken into consideration.
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Objective To investigate the effects of sulforaphane (SFN) on proliferation and invasion of human small cell lung cancer NCI-H446 and the activity of matrix metalloproteinase (MMP) -9.Methods NCI-H446 cells were cultured with 0,25,50,100 μmol/L SFN for 24 ~ 72 h,then MTT assay was employed to detect cell proliferation.Chamber invasion assay was used to study the cell invasion,and gelatin zymography assay was implied in MMP-9 enzyme activity.Results After treatment of 25,50,100μmol/L SFN,the growth of NCI-H446 cells were inhibited.When cells were incubated with 25,50,100μmol/L of SFN for 72h,the inhibition ratio was ( 11.1 ± 2.26 ) %,( 25.2 ± 3.24 ) % and ( 44.6 ±4.2) %,respectively,the difference was statistically significant compared with the solvent control group ( t =10.685,8.417,5.264,P <0.05 ).Chamber invasion assay showed that NCI-H446 cell invasion could be reduced.25,50,100 μmol/L of SFN could decrease the trans-membrane cells to (48.6 ± 1.84)%,(35.4 ± 2.22) % and (27.8 ± 1.36) %,and it was statistically significant compared with the solvent control group ( t =6.341,5.562,4.925,P <0.05 ),respectively.In addition,MMP-9 activity was significantly inhibited by SFN.25,50,100 μmol/L of SFN could decrease the gray value of MMP-9 to 764 ±18.4,685 ± 14.74 and 638 ± 21.54 ( control group 822 ± 12.53,t =4.971,7.582,11.235,respectively,P <0.05).Conclusions SFN can inhibit NCI-H446 cells growth,invasion and the activity ofMMP-9.
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<p><b>OBJECTIVE</b>To explore the efficacy of homemade hemostatics of injected gelatin matrix (HIGM) for immediately treating blunt hepatic trauma in canine model without additional pressure.</p><p><b>METHODS</b>A total of 27 commercial hybrid dogs underwent celiotomy to establish hepatic trauma model after general anesthesia. The dogs were prospectively randomized into 3 groups: the treatment group (n=9, with the direct application of homemade hemostat), the positive control group (n=9, with thrombin solution), and the negative control group (n=9, with 0.9% normal saline). Time to hemostasis and intra-abdominal blood loss were recorded, and heart rate (HR), mean arterial pressure (MAP), and hematological parameters were compared among these three groups. Gross examinations were performed 30 minutes after surgery.</p><p><b>RESULTS</b>Significantly shorter time to hemostasis [(1.20±0.33) min] and less blood loss [(47.22±8.61) ml] were observed in the treatment group than in control groups (P 0.05). No cases of bleeding occurred in any animals in the treatment group, and no signs of infection and adhesion formation were evident due to exposure to HIGM. Two cases in the positive control group (22.22%) were found to have rebleeding. All animals in the negative control group experienced visible bleeding.</p><p><b>CONCLUSION</b>HIGM is effective for controlling bleeding after hepatic trauma without the additional compression, and therefore may be valuable in field surgery.</p>
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Animales , Perros , Modelos Animales de Enfermedad , Gelatina , Hemostáticos , Inyecciones , Hígado , Heridas y LesionesRESUMEN
The expressions of estrogen receptor (ER) α,ERβ1,and ERβ2 in thyroid carcinoma,thyroid adenoma,and adjacent normal thyroid tissue were determined by RT-PCR and immunohistochemistry,and the relationship of expression levels of three ERs and the main clinical pathologic parameters were evaluated.The result showed that ERα mRNA expression in papillary thyroid carcinoma(PTC) tissues was significantly higher than adjacent normal tissue,while,ERβ2 mRNA expression in PTC tissues was significantly lower than normal tissue ( P<0.05 ).The positive expression of ERβ1 and ERβ2 in the thyroid carcinoma was related with the histological variant of carcinomas; the ERβ2 expressions were related with the lymph node metastasis and TNM staging of the carcinomas.These results suggest that the decreased ERβ2 expression may induce proliferation and carcinogenesis of thyroid follicular epithelial cells as well as progression of the carcinoma,and is a powerful prognostic indicator.
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Sedum alfredii Hance has been identified as zinc (Zn) and cadmium (Cd) co-hyperaccumulator. In this paper the relationships of Zn or Cd hyperaccumulation to the generation and the role of H2O2 in Sedum alfredii H. were examined. The results show that Zn and Cd contents in the shoots of Sedum alfredii H. treated with 1000 micromol/L Zn2+ and/or 200 micromol/L Cd2+ increased linearly within 15 d. Contents of total S, glutathione (GSH) and H2O2 in shoots also increased within 15 d, and then decreased. Total S and GSH contents in shoots were higher under Cd2+ treatment than under Zn2+ treatment. However, reverse trends of H2O2 content in shoots were obtained, in which much higher H2O2 content was observed in Zn2+-treated shoots than in Cd2+-treated shoots. Similarly, the microscopic imaging of H2O2 accumulation in leaves using H2O2 probe technique showed that much higher H2O2 accumulation was observed in the Zn2+-treated leaf than in the Cd2+-treated one. These results suggest that there are different responses in the generation of H2O2 upon exposure to Zn2+ and Cd2+ for the hyperaccumulator Sedum alfredii H. And this is the first report that the generation of H2O2 may play an important role in Zn hyperaccumulation in the leaves. Our results also imply that GSH may play an important role in the detoxification of dissociated Zn/Cd and the generation of H2O2.
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Cadmio , Farmacología , Glutatión , Metabolismo , Peróxido de Hidrógeno , Metabolismo , Cinética , Hojas de la Planta , Metabolismo , Brotes de la Planta , Sedum , Metabolismo , Azufre , Metabolismo , Zinc , FarmacologíaRESUMEN
<p><b>BACKGROUND</b>The chronic pathological changes in vascular walls of hypertension may exert destructive effects on multiple organ systems. Accumulating evidence indicates that inflammatory reactions are involved in the pathological changes of hypertension. Three peroxisome proliferator-activated receptors (PPARs) have been identified: PPARalpha, PPARbeta/delta, and PPARgamma, all of which have multiple biological effects, especially the inhibition of inflammation. The aim of this study was to evaluate PPAR isoforms expression profile in important organs of spontaneously hypertensive rats (SHR) and to understand the modulation of endogenous PPAR isoforms under inflammatory condition.</p><p><b>METHODS</b>Tissues (kidney, liver, heart, and brain) were dissected from SHR and age-matched control Wistar-Kyoto rats (WKY) to investigate the abundance of PPAR isoforms and PPAR-responsive genes (acyl-CoA oxidase and CD36). The expression of CCAAT/enhancer-binding protein delta (C/EBPdelta), which can trans-activate PPARgamma expression, was also observed. The inflammatory response was analyzed by the expression of inflammatory mediators inducible nitric oxide synthase (iNOS), intercellular adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1), E-selectin, interleukin-1 beta (IL-1beta), and tumor necrosis factor alpha (TNFalpha), and formation of carbonyl and nitrated proteins.</p><p><b>RESULTS</b>The expressions of 3 PPAR isoforms and PPAR-responsive genes were markedly upregulated in SHR compared with those of WKY. Specifically, the expression of PPARalpha protein in the kidney, liver, heart and brain increased by 130.76%, 91.48%, 306.24%, and 90.70%; PPARbeta/delta upregulated by 109.34%, 161.98%, 137.04%, and 131.66%; PPARgamma increased by 393.76%, 193.17%, 559.29%, and 591.18%. In consistent with the changes in PPARgamma, the expression of C/EBPdelta was also dramatically elevated in SHR. Inflammatory mediators expressions were significantly increased in the most organs of SHR than WKY. As a consequence, increased formation of carbonyl and nitrated proteins were also observed in the most organs of SHR.</p><p><b>CONCLUSIONS</b>These findings suggest an enhanced inflammatory response in the organs of SHR, which might play a key role in pathogenesis of hypertension and secondary organ complications. Changes (increases) in PPARs expression may reflect a compensatory mechanism to the inflammatory status of hypertensive rats.</p>
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Animales , Masculino , Ratas , Presión Sanguínea , Western Blotting , Selectina E , Genética , Metabolismo , Expresión Génica , Hipertensión , Genética , Metabolismo , Inflamación , Genética , Metabolismo , Interleucina-1beta , Genética , Metabolismo , PPAR alfa , Genética , Metabolismo , PPAR delta , Genética , Metabolismo , PPAR gamma , Genética , Metabolismo , Receptores Activados del Proliferador del Peroxisoma , Genética , Metabolismo , Pletismografía , Métodos , Ratas Endogámicas SHR , Ratas Endogámicas WKY , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Factor de Necrosis Tumoral alfa , Genética , Metabolismo , Molécula 1 de Adhesión Celular Vascular , Genética , MetabolismoRESUMEN
<p><b>OBJECTIVE</b>To screen proteins interacting with HCV NS4A protein in leukocytes by yeast-double hybridization.</p><p><b>METHODS</b>The bait plasmid pGBKT7-NS4A was transformed into yeast AH109 was transformed, and the expressing of the fusion protein was identified by SDS-page. The transformed yeast was mated with yeast Y187 containing leukocytes cDNA library plasmid in 2xYPDA medium. Diploid yeast was plated on synthetic dropout nutrient medium (SD/-Trp-Leu-His-Ade) and synthetic dropout nutrient medium (SD/-Trp-Leu-His-Ade) containing x-alpha-gal for selecting two times and screening. After extracting and sequencing of plasmid from blue colonies, analysis was conducted by bioinformatics. And, the gene encoding the interesting protein was cloned, and back-cross was performed.</p><p><b>RESULTS</b>Forty-five colonies were sequenced, among them, 29 colonies were human calcium modulating cyclophilin ligand (CAML). The gene encoding CAML was cloned, and the interaction between NS4A and CAML was ensured.</p><p><b>CONCLUSION</b>Seven kinds of proteins interacting with NS4A in leukocytes were successfully screened and the results brought some new clues for studying the pathogenesis of HCV.</p>
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Humanos , Proteínas Adaptadoras Transductoras de Señales , Genética , Metabolismo , Proteínas Portadoras , Genética , Metabolismo , Clonación Molecular , Biblioteca de Genes , Leucocitos , Biología Celular , Metabolismo , Unión Proteica , Transformación Genética , Técnicas del Sistema de Dos Híbridos , Proteínas no Estructurales Virales , Proteínas Virales , Genética , MetabolismoRESUMEN
<p><b>OBJECTIVE</b>To provide a sound cell source for further ex-vivo gene therapy for chronic pain, we attempt to develop an immortalized rat astrocyte cell line that expresses enkephalin regulated by doxycycline.</p><p><b>METHODS</b>Retrovirus infection method was employed to develop an immortalized rat astrocyte cell line that could express enkephalin regulated by doxycycline. The hPPE gene expression level of immoralized astroyte cells (IAC)/ hPPE was detected by RT-PCR, indirect immunofluorescence staining and radioimmunoassay.</p><p><b>RESULTS</b>IAC carrying Tet-on system transfected with preproenkephalin gene could secrete enkephalin that was regulated by doxycycline in a dose-dependent manner and hPPE gene activation could be repeated in on-off-on cycles through administration or removal of doxycycline.</p><p><b>CONCLUSION</b>An immortalized rat astrocyte cell line that secrete enkephalin under the control of doxycycline is established successfully, which provides a research basis for transgenic cell transplantation for analgesia.</p>
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Animales , Ratas , Antibacterianos , Farmacología , Astrocitos , Línea Celular Transformada , Enfermedad Crónica , Doxiciclina , Farmacología , Encefalinas , Genética , Metabolismo , Terapia Genética , Métodos , Vectores Genéticos , Neurotransmisores , Genética , Metabolismo , Manejo del Dolor , Precursores de Proteínas , Genética , Metabolismo , Retroviridae , GenéticaRESUMEN
<p><b>BACKGROUND</b>NR2B containing N-methyl-D-aspartate (NMDA) receptor plays an important role in the facilitation and maintenance of neuropathic pain. The discrete distribution of NR2B subunit in the central nervous system (CNS) may support reduced side effects of agents that act selectively at this site. Therefore, we investigated the hypothesis that a humoral autoimmune response targeting the NR2B subunit of NMDA receptor relieves pain like behaviours produced by peripheral injury.</p><p><b>METHODS</b>Rats were immunized subcutaneously with NR2B-Keyhole Limpet Hemocyanin (NR2B-KLH) three times at two-week intervals. NR2B specific IgG titres in sera and cerebrospinal fluid were determined by indirect ELISA. Seven days after the third immunization, 2 of the 3 terminal branches of the sciatic nerve (tibial and common peroneal nerves) were tightly ligated. Behavioural testing was carried out on every other day after surgery, until 7 days after surgery. The lumbar spinal cord (L4-6) was removed on day 7 after ligation. The expression of NR2B protein in the lumbar spinal cord was determined using Western blotting.</p><p><b>RESULTS</b>After the second vaccination, NR2B specific IgG in sera was detected to be > 0.5 microg/ml in six of nine rats. After the third vaccination, all the immunized rats had > 2.2 microg/ml. Titres of NR2B specific IgG in sera peaked 42 days post initial immunization and persisted for over 70 days. No NR2B specific IgG was detected in sera from PBS or KLH group. The behavioural thresholds in NR2B group were significantly higher than those in PBS and KLH groups on day 7 after ligation. NR2B specific IgG in CSF in NR2B group could not be detected on day 1 before spinal dissection; but could be detected on day 7 after surgery. The expression of NR2B protein in group NR2B was significantly lower than in PBS and KLH groups on day 7 after surgery.</p><p><b>CONCLUSION</b>The NR2B peptide could be used as a vaccine against neuropathic pain, which could be associated with reduction of NR2B protein in the lumbar spinal cord.</p>
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Animales , Femenino , Ratas , Adyuvantes Inmunológicos , Western Blotting , Modelos Animales de Enfermedad , Ensayo de Inmunoadsorción Enzimática , Hemocianinas , Alergia e Inmunología , Inmunoglobulina G , Alergia e Inmunología , Neuralgia , Alergia e Inmunología , Dimensión del Dolor , Ratas Sprague-Dawley , Receptores de N-Metil-D-Aspartato , Alergia e Inmunología , Metabolismo , Proteínas Recombinantes de Fusión , Alergia e Inmunología , Médula Espinal , Metabolismo , Factores de Tiempo , Vacunas , Alergia e InmunologíaRESUMEN
The hypertension is one of chronic vascular diseases, which often implicates multiple tissues causing stroke, cardiac hypertrophy, and renal failure. A growing body of evidence suggests that inflammatory mechanisms are important participants in the pathophysiology of hypertension. In this study, the inflammatory status of these tissues (kidney, liver, heart, and brain) in spontaneously hypertensive rats (SHR) was analyzed and its molecular mechanism was explored. The tissues were dissected from SHR and age-matched control Wistar-Kyoto (WKY) rats to investigate the abundance of inflammation-related mediators (IL-1beta, TNFalpha, ICAM-1, iNOS, C/EBPdelta and PPARgamma). mRNA levels were determined by reverse transcription-polymerase chain reaction and protein expression was evaluated by Western blot. To evaluate the oxidative stress of tissues, carbonyl protein content and total antioxidant capacity of tissues were detected by spectrophotometry and ferric reduction ability power (FRAP) method. The results suggest that: (1) Expressions of inflammation-related mediators (IL-1beta, TNFalpha, ICAM-1, iNOS, C/EBPdelta and PPARgamma) in SHR were higher compared with those in WKY rats except no evident increase of IL-1beta mRNA in liver and brain in SHR. (2) Tissues in SHR contained obviously increased carbonyl protein (nmol/mg protein) compared to that in WKY rats (8.93+/-1.08 vs 2.27+/-0.43 for kidney, 2.23+/-0.23 vs 0.17+/-0.02 for heart, 13.42+/-1.10 vs 5.72+/-1.01 for brain, respectively, P<0.05). However, no evident difference in the amount of carbonyl protein in liver was detected between SHR and WKY rats. (3) Total antioxidant capacities of kidney, liver, heart and brain were markedly lower in SHR than that in WKY rats (P<0.05). Thus, the present data reveal a higher inflammatory status in the important tissues in SHR and indicate that inflammation might play a potential role in pathogenesis of hypertension and secondary organ complications.
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Animales , Masculino , Ratas , Encéfalo , Metabolismo , Patología , Citocinas , Genética , Metabolismo , Hipertensión , Patología , Inflamación , Patología , Interleucina-1beta , Genética , Metabolismo , Riñón , Metabolismo , Patología , Miocardio , Metabolismo , Patología , Estrés Oxidativo , Alergia e Inmunología , ARN Mensajero , Genética , Metabolismo , Ratas Endogámicas SHR , Ratas Endogámicas WKY , Factor de Necrosis Tumoral alfa , Genética , MetabolismoRESUMEN
Elsholtzia argyi and Elsholtzia splendens, which are Chinese endemic Pb/Zn mined and Cu mined ecotype respectively, were investigated in the aspect of their response to Pb toxicity in the presence or absence of EDTA addition. After 8 d's Pb treatment, root length, root surface area and root volume of E. splendens decreased much more than those of E. argyi, and reduced considerably with increase of Pb, while no marked change was noted for root average diameter. Compared to E. argyi, length of root with diameter (D)<0.2 mm was significantly reduced for E. splendens as Pb increasing. Root with cross-sectional area of D<0.1 mm for E. splendens was at Pb> or =10 mg/L, while for E. argyi, it was at Pb> or =25 mg/L. DW of E. splendens decreased much more than that of E. argyi with increase of Pb. E. argyi exhibited much more tolerance to Pb toxicity than E. splendens. Treatment with 100 mg/L Pb plus 50 mmol/L EDTA significantly decreased the length and surface area of D< or =0.2 mm root, increased the length and surface area of 0.2< or =D< or =0.8 mm root for the case of E. argyi, while for E. splendens, length and surface area of D<0.6 mm root reduced, as compared to 100 mg/L Pb treatment, alone. At 100 mg/L Pb, shoot Pb accumulation in E. splendens and E. argyi were 27.9 and 89.0 microg/plant DW respectively, and much more Pb was uptaken by the root and translocated to the stem of E. argyi as compared to E. splendens. Treatment of the plant with 100 mg/L Pb plus 50 mmol/L EDTA increase leaf Pb accumulation from 16.8 to 84.9 g/plant for E.splendens and from 18.8 to 52.5 g/plant for E. argyi, while both root and stem Pb pronouncedly reduced for both Elsholtzia species. The increased translocation of Pb to the leaf of E. splendens than that of E. argyi at the treatment of 100 mg/L Pb plus 50 mmol/L EDTA should be further investigated.
Asunto(s)
Biodegradación Ambiental , Proliferación Celular , Relación Dosis-Respuesta a Droga , Residuos Industriales , Lamiaceae , Clasificación , Metabolismo , Plomo , Farmacocinética , Toxicidad , Raíces de Plantas , Metabolismo , Especificidad de la EspecieRESUMEN
Copper accumulation and intracellular distribution in Elsholtzia splendens, a native Chinese Cu-tolerant and accumulating plant species, was investigated by transmission electron microscope (TEM) and gradient centrifugation techniques. Copper concentrations in roots, stems and leaves of E. splendens increased with increasing Cu levels in solution. After exposure to 500 micromol/L Cu for 8 d, about 1000 mg/kg Cu were accumulated in the stem and 250 mg/kg Cu in the leaf of E. splendens. At 50 micromol/L Cu, no significant toxicity was observed in the chloroplast and mitochondrion within its leaf cells, but separation appeared at the cytoplasm and the cell wall within the root cells. At >250 micromol/L Cu, both root and leaf cell organelles in E. splendens were damaged heavily by excessive Cu in vivo. Copper subcellular localization in the plant leaf after 8 days' exposure to 500 micromol/L Cu using gradient centrifugation techniques was found to be decreased in the order: chloroplast>cell wall>soluble fraction>other organelles. The plant root cell wall was found to be the site of highest Cu localization. Increase of Cu exposure time from 8 d to 16 d, increased slightly Cu concentration in cell wall fraction in roots and leaves, while that in the chloroplast fraction decreased in leaves of the plants grown in both 0.25 micromol/L and 500 micromol/L Cu. TEM confirmed that much more Cu localized in cell walls of E. splendens roots and leaves, but also more Cu localized in E. splendens' chloroplast when the plant is exposed to Cu levels>250 micromol/L, as compared to those in the plant grown in 0.25 micromol/L Cu. Copper treatment at levels>250 micromol/L caused pronounced damage in the leaf chloroplast and root organelles. Copper localization in cell walls and chloroplasts could mainly account for the high detoxification of Cu in E. splendens.
Asunto(s)
Cobre , Metabolismo , Lamiaceae , Biología Celular , Metabolismo , Microscopía Electrónica de Transmisión , Hojas de la Planta , Biología Celular , Metabolismo , Raíces de Plantas , Biología Celular , Metabolismo , Tallos de la Planta , Biología Celular , MetabolismoRESUMEN
A solution with different Cu supply levels was cultured to investigate gama-aminobutyric acid (GABA) accumulation in Elsholtzia splendens, a native Chinese Cu-tolerant and accumulating plant species. Increasing Cu from 0.25 to 500 micromol/L significantly enhanced levels of GABA and histidine (His), but considerably decreased levels of aspartate (Asp) and glutamate (Glu) in the leaves. The leaf Asp level negatively correlated with leaf Cu level, while leaf GABA level positively correlated with leaf Cu level. The leaf Glu level negatively correlated with leaf GABA level in Elsholtzia splendens. The depletion of leaf Glu may be related to the enhanced synthesis of leaf GABA under Cu stress.