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1.
Anim Genet ; 54(6): 803-807, 2023 Dec.
Article in English | MEDLINE | ID: mdl-37705287

ABSTRACT

Semen is a measure of the reproductive efficiency of roosters, which affects the economic benefits of white-feathered broilers. Over the years, research in this field has mainly focused on hens, while there have been fewer studies on the reproductive traits of roosters. To identify the genes related to the semen traits of roosters, we used a chicken 55 K SNP chip to genetically type the white-feathered population (220) and performed imputation with resequencing data from 97 roosters. In total, 1 048 576 SNPs were obtained and used for genome-wide association analysis of semen volume, from which 197 genome-wide significant markers were identified, all within the interval of 13.82-16.12 Mb on chromosome 7. By combining our results with the biological functions of genes in the interval, four candidate genes were identified that potentially relate to semen volume: FAPP1, OSBPL6, SESTD1 and SSFA2. Our findings may provide a basis for further research on the genetic mechanism and marker-assisted selection of semen volume in white-feathered broilers.


Subject(s)
Chickens , Genome-Wide Association Study , Animals , Male , Female , Genome-Wide Association Study/veterinary , Chickens/genetics , Semen , Semen Analysis , Phenotype , Polymorphism, Single Nucleotide
2.
Zhonghua Yi Xue Za Zhi ; 103(19): 1455-1460, 2023 May 23.
Article in Chinese | MEDLINE | ID: mdl-37198107

ABSTRACT

Objective: To investigate the anatomic zone localization based on biparametric magnetic resonance imaging (bpMRI) for the prediction of the risk degree in patients with prostate cancer. Methods: A total of 92 patients with prostate cancer confirmed by radical surgery in First Affiliated Hospital, Air Force Medical University, from January 2017 to December 2021 were collected. All patients underwent bpMRI (non-enhanced scan and DWI). According to ISUP grade, those patients were divided into low-risk group [≤grade 2, n=26, aged 71 (64.0, 5.2) years] and high-risk group[≥grade 3, n=66, aged 70.5 (63.0, 74.0) years]. The interobserver consistency test for ADC values was evaluated using the intraclass correlation coefficients (ICC). The differences in total prostate specific antigen (tPSA) between the two groups were compared and the χ2 test was used to compare the differences in the risk of prostate cancer in the transitional and peripheral zone. Independent correlation factors for prostate cancer risk were analyzed by logistic regression using high and low risk of prostate cancer as dependent variables, including factors such as anatomical zone, tPSA, apparent diffusion coefficient mean (ADCmean), apparent diffusion coefficient minimum (ADCmin) and age. Receiver operating characteristic (ROC) curves were plotted to assess the efficacy of the combined models of anatomical zone, tPSA, and anatomical partitioning+tPSA for diagnosing prostate cancer risk. Results: The ICC values of the ADCmean and ADCmin between the observers were 0.906 and 0.885, respectively, with good agreement. The tPSA in the low-risk group was lower than that in the high-risk group [19.64 (10.29, 35.18) ng/ml vs 72.42 (24.79, 187.98) ng/ml; P<0.001]; the risk of prostate cancer in the peripheral zone was higher than that in the transitional zone, and the difference was statistically significant (P<0.01). Multifactorial regression showed that anatomical zones (OR=0.120, 95%CI:0.029-0.501, P=0.004) and tPSA (OR=1.059, 95%CI:1.022-1.099, P=0.002) were risk factors for prostate cancer risk. The diagnostic efficacy of the combined model (AUC=0.895, 95%CI: 0.831-0.958) was better than the predictive efficacy of the single model for both anatomical partitioning (AUC=0.717, 95%CI:0.597-0.837) and tPSA (AUC=0.801, 95%CI: 0.714-0.887) (Z=3.91, 2.47; all P<0.05). Conclusions: The malignant degree of prostate cancer in peripheral zone was higher than that in transitional zone. Combination of anatomic zone located by bpMRI and tPSA can be used to predict the risk of prostate cancer before surgery, expected to provide support for patients to develop personalized treatment strategies.


Subject(s)
Prostatic Neoplasms , Male , Humans , Prostatic Neoplasms/diagnosis , Magnetic Resonance Imaging/methods , Prostate-Specific Antigen , Diffusion Magnetic Resonance Imaging/methods , ROC Curve , Retrospective Studies
3.
Zhonghua Gan Zang Bing Za Zhi ; 28(4): 338-344, 2020 Apr 20.
Article in Chinese | MEDLINE | ID: mdl-32403887

ABSTRACT

Objective: To observe the curative effects of berberine in rats with high-fat diet induced non-alcoholic fatty liver and to further explore its possible mechanism. Methods: Twenty-six Sprague-Dawley rats (120-160 g) were randomly divided into 3 groups: control group (n = 8), model group (n = 10) and treatment group (n = 8). Rats in the control group were fed with regular diet, and the model group and the treatment group were fed a high-fat diet. At the 12th week, two rats in the in the model group were sacrificed to verify whether model was successful established. Subsequently, treatment group rats were given a gavage of berberine at a dose of 150 mg·kg(-1)·d(-1) for 4 weeks, and the control and the model group rats were given the same dose of normal saline. Rats were sacrificed at week 16th. HE staining was used to observe the changes in the intestinal mucosa of rats. Sudan black B staining was used to observe the fatty changes in liver. Immunohistochemical staining was used to observe the expression level of occludin protein in the intestinal epithelium. A real-time 16S rDNA PCR method was used to measure the number of escherichia coli, bacteroides and faecalibacterium prausnitzii in the feces of rats. Results: Model group had a higher serum levels of endotoxin (0.288 ± 0.045) and tumor necrosis factor (TNF)-α (1.07 ± 0.11) than the control group (0.192 ± 0.049, 0.94 ± 0.07) (P < 0.05). Berberine intervention had significantly reduced endotoxin (0.213 ± 0.025) and TNF-α level (0.93 ± 0.07) (P < 0.05). The expression level of occludin protein was significantly lower in the intestinal mucosa of model group than that of control group (0.166 ± 0.014), and berberine had promoted the expression of occludin protein in intestinal mucosa (0.055 ± 0.009), but the difference was not statistically significant (P > 0.05). At the same time, compared with the model group (7.29 ± 0.47), the number of bacteroidetes in the control group (9.49 ± 0.59) was decreased, while the number of bacteroidetes in the treatment group was increased (9.77 ± 0.87). The number of escherichia coli (6.92 ± 0.77) and faecalibacterium prausnitzii (8.70 ± 0.62) in the model group were increased than control group (5.42 ± 0.63, 9.49 ± 0.59), while the number of escherichia coli (6.34 ± 0.71) and faecalibacterium prausnitzii (9.77 ± 0.87) (P < 0.05) was reduced with the intervention of berberine. Conclusion: Berberine could effectively protect the intestinal barrier function in rats with NAFLD and the possible mechanism of action behind it may be the regulation of intestinal flora.


Subject(s)
Berberine/therapeutic use , Non-alcoholic Fatty Liver Disease/drug therapy , Animals , Diet, High-Fat , Liver , Random Allocation , Rats , Rats, Sprague-Dawley
6.
Nat Commun ; 6: 8014, 2015 Aug 14.
Article in English | MEDLINE | ID: mdl-26272722

ABSTRACT

The distribution of cations in Li-ion battery cathodes as a function of cycling is a pivotal characteristic of battery performance. The transition metal cation distribution has been shown to affect cathode performance; however, Li is notoriously challenging to characterize with typical imaging techniques. Here laser-assisted atom probe tomography (APT) is used to map the three-dimensional distribution of Li at a sub-nanometre spatial resolution and correlate it with the distribution of the transition metal cations (M) and the oxygen. As-fabricated layered Li1.2Ni0.2Mn0.6O2 is shown to have Li-rich Li2MO3 phase regions and Li-depleted Li(Ni0.5Mn0.5)O2 regions. Cycled material has an overall loss of Li in addition to Ni-, Mn- and Li-rich regions. Spinel LiNi0.5Mn1.5O4 is shown to have a uniform distribution of all cations. APT results were compared to energy dispersive spectroscopy mapping with a scanning transmission electron microscope to confirm the transition metal cation distribution.

7.
Genet Mol Res ; 14(4): 18650-61, 2015 Dec 29.
Article in English | MEDLINE | ID: mdl-26782515

ABSTRACT

Recombinant adenovirus vector systems have been used extensively in protein research and gene therapy. However, the construction and characterization of recombinant adenovirus is a tedious and time-consuming process. TIGIT is a recently discovered immunosuppressive molecule that plays an important role in maintaining immunological balance. The construction of recombinant adenovirus mediating TIGIT expression must be simplified to facilitate its use in the study of TIGIT. In this study, the TIGIT gene was combined with green fluorescent protein (GFP); the TIGIT-GFP gene was inserted into a gateway plasmid to construct a TIGIT-GFP adenovirus. HEK 293A cells were infected with the adenovirus, which was then purified and subjected to virus titering. TIGIT-GFP adenovirus was characterized by flow cytometry and immunofluorescence, and its expression in mouse liver was detected by infection through caudal vein injection. The results showed the successful construction of the TIGIT-GFP adenovirus (5 x 10(10) PFU/mL). Co-expression of TIGIT and GFP was identified in 293A and liver cells; synthesis and positioning of TIGIT-GFP was viewed under a fluorescence microscope. TIGIT-GFP was highly expressed on liver cells 1 day (25.53%) after infection and faded 3 days (11.36%) after injection. In conclusion, the fusion of TIGIT with GFP allows easy, rapid, and uncomplicated detection of TIGIT translation. The construction of a TIGIT-GFP adenovirus, mediating TIGIT expression in vitro and in vivo, lays the foundation for further research into TIGIT function and gene therapy. Moreover, the TIGIT-GFP adenovirus is a helpful tool for studying other proteins (which could replace the TIGIT gene).


Subject(s)
Adenoviridae/genetics , Genetic Vectors/genetics , Green Fluorescent Proteins/genetics , Receptors, Immunologic/genetics , Recombinant Fusion Proteins/genetics , Animals , Cell Line , Gene Expression , Gene Order , Genetic Vectors/isolation & purification , Hepatocytes/metabolism , Humans , Mice , Transduction, Genetic
8.
Tech Coloproctol ; 16(4): 277-83, 2012 Aug.
Article in English | MEDLINE | ID: mdl-22527922

ABSTRACT

BACKGROUND: Invasion of submucosa (ISM) is required for the pathological diagnosis of colorectal cancer according to the WHO criteria. A large proportion of colorectal cancers may be underdiagnosed as high-grade intraepithelial neoplasia (HGIN) because ISM is not identified in the preoperative biopsy. The aim of this study was to investigate the clinicopathologic features that are associated with missing the diagnosis of ISM in biopsy specimens of invasive colorectal cancer. METHODS: Three hundred and sixteen patients diagnosed with colorectal cancer between January 2007 and December 2008 with well-preserved preoperative biopsy specimens were enrolled in the study. Three hundred and eleven patients had an isolated lesion, and five had two lesions. Biopsy specimens were reevaluated by two senior pathologists. Clinicopathologic features, biopsy pathology and surgical pathology results of all patients were analyzed by univariate and multivariate analyses. RESULTS: ISM was identified in 216 cases (67.3 %) by biopsy-based pathological examination, and missed in 105 (32.7 %) cases, 72 of which were diagnosed as HGIN. Univariate analysis indicated that in colorectal cancer patients with smaller biopsy specimens (P = 0.042), mucinous or signet-ring cell carcinoma (P = 0.003), higher WHO tumor grade (P = 0.001) and positive lymph nodes (P = 0.011), ISM was more likely to be missed. There was a trend toward an increased diagnosis of ISM with the increase in the number of biopsy specimens (P = 0.105). On multivariate logistic regression analysis, smaller biopsy specimens (OR, 1.810; 95 % CI, 1.081-3.032; P = 0.024) and higher WHO tumor grade (OR, 2.073; 95 % CI, 1.046-4.107; P = 0.037) were the only factors associated with failure to identify ISM. CONCLUSIONS: A large number of invasive colorectal cancers are at risk of being underdiagnosed as HGIN by biopsy-based pathology. The smaller the biopsy size, the less likely it is that the muscularis mucosae is included in the specimen. Also, in the more advanced or aggressive colorectal cancers, ISM is more likely to be missed on biopsy, which may be due to the destruction of the muscularis mucosae by more aggressive cancers.


Subject(s)
Carcinoma in Situ/pathology , Colorectal Neoplasms/pathology , Adult , Aged , Aged, 80 and over , Biopsy , Chi-Square Distribution , Diagnosis, Differential , Female , Humans , Incidence , Logistic Models , Lymphatic Metastasis , Male , Middle Aged , Neoplasm Grading , Neoplasm Invasiveness , Retrospective Studies
9.
Diabetologia ; 55(3): 801-11, 2012 Mar.
Article in English | MEDLINE | ID: mdl-22130579

ABSTRACT

AIMS/HYPOTHESIS: Increased renal mast cells have been detected in diabetic nephropathy. However, only a few patients have been examined. Evidence of the involvement of mast cells in diabetic nephropathy is still scarce, and no observation of mast cells during the development of diabetic nephropathy has yet been reported in humans. Here, we examined changes in renal mast cells in patients at different stages of diabetic nephropathy and related these to the development of the disease. METHODS: Eighty patients at different clinical stages of diabetic nephropathy and 16 normal kidney donors were recruited. Immunohistochemical staining for tryptase, chymase, TGF-ß1, renin and TNF-α was done on renal sections from patients and control participants. Changes in mast cell number, degranulation, subtype and phenotype were examined. Correlation between mast cells and patients' clinical and pathological indices was analysed. RESULTS: With progression of diabetic nephropathy, the number and degranulation level of mast cells increased. Increase in mast cell number and degranulation level correlated significantly with tubular interstitial injury. Almost all renal mast cells in patients with diabetic nephropathy were found to produce chymase, renin, TGF-ß1 and TNF-α. The level of TNF-α in mast cells increased with progression of diabetic nephropathy. CONCLUSIONS/INTERPRETATION: This study suggests that mast cells are involved in development of diabetic nephropathy. Through release of bioactive substances, such as tryptase, chymase, TGF-ß1, renin and TNF-α, into the tubular interstitium by degranulation, mast cells could promote renal inflammation and fibrosis, and thus contribute to diabetic nephropathy.


Subject(s)
Diabetic Nephropathies/immunology , Diabetic Nephropathies/physiopathology , Kidney/pathology , Kidney/physiopathology , Mast Cells/immunology , Adult , Albuminuria/etiology , Albuminuria/physiopathology , Cell Count , Cell Degranulation , Diabetic Nephropathies/pathology , Diabetic Nephropathies/urine , Disease Progression , Female , Fibrosis , Humans , Hypertension/etiology , Hypertension/physiopathology , Immunohistochemistry , Kidney/immunology , Kidney Tubules/immunology , Kidney Tubules/metabolism , Kidney Tubules/pathology , Kidney Tubules/physiopathology , Male , Mast Cells/pathology , Mast Cells/physiology , Middle Aged , Proteinuria/etiology , Proteinuria/physiopathology , Renal Insufficiency/etiology , Renal Insufficiency/physiopathology , Severity of Illness Index , Tumor Necrosis Factor-alpha/metabolism
10.
Transplant Proc ; 43(10): 3690-3, 2011 Dec.
Article in English | MEDLINE | ID: mdl-22172827

ABSTRACT

OBJECTIVE: The aim was to investigate the factors that affect graft function at 2 years after transplantation in living related-donor kidney transplantation. METHODS: We retrospectively reviewed the clinical data of 144 patients who underwent living related-donor kidney transplantation in our hospital from December 2005 to December 2008. Recipients were divided into 2 groups according to glomerular filtration rate (GFR) at 2 years after transplantation: ≤60 mL/min/1.73 m2 (n=51) and >60 mL/min/1.73 m2 (n=93). Variables which affected graft function were compared between the groups. The significant factors were analyzed using logistic regression. RESULTS: Univariate analysis showed significant differences for donor age, donor GFR, recipient weight, recipient body mass index, donor-to-recipient body weight ratio, and acute rejection episodes (P<.05). Logistic regression analysis revealed the independent factors affecting renal function at 2 years after transplantation to be donor GFR (ß=0.032; odds ratio [OR] 1.032; P=.004) and recipient body weight (ß=-0.069; OR 0.934; P=.001). Receiver operating characteristic (ROC) curve analysis showed cutoff values of donor GFR and recipient body weight to be >111.25 mL/min/1.73 m2, and ≤67 kg, respectively. Areas under the ROC curve of donor GFR and recipient body weight were 0.612 and 0.665, respectively. Sensitivity and specificity of donor GFR were 43.0% and 78.4%, respectively. Sensitivity and specificity of recipient body weight were 82.8% and 45.1%, respectively. CONCLUSIONS: Donor GFR and recipient body weight were the independent factors effecting renal function at 2 years after transplantation.


Subject(s)
Graft Survival , Kidney Transplantation , Living Donors , Adult , Body Weight , Chi-Square Distribution , China , Female , Glomerular Filtration Rate , Humans , Kidney Transplantation/adverse effects , Living Donors/statistics & numerical data , Logistic Models , Male , Middle Aged , Odds Ratio , ROC Curve , Retrospective Studies , Risk Assessment , Risk Factors , Time Factors , Treatment Outcome
11.
Br J Pharmacol ; 153(7): 1456-64, 2008 Apr.
Article in English | MEDLINE | ID: mdl-18264122

ABSTRACT

BACKGROUND AND PURPOSE: Rhein, an anthraquinone compound isolated from rhubarb, has been proved effective in treatment of experimental diabetic nephropathy (DN). To explore the mechanism of its therapeutic effect on DN, rhein was tested for its effect on the hexosamine pathway. EXPERIMENTAL APPROACH: The influence of rhein on cellular hypertrophy, fibronectin synthesis, glucose uptake, glutamine: fructose 6-phosphate aminotransferase (GFAT) activity, UDP-N-acetylglucosamine (UDP-GlcNAc) level and TGF-beta1 and p21 expression was evaluated in MCGT1 cells, a GLUT1 transgenic rat mesangial cell line. GFAT activity in normal rat mesangial cells in high glucose concentrations and in vitro was also measured. KEY RESULTS: Significantly increased fibronectin synthesis, cellular hypertrophy, much higher GFAT activity and UDP-GlcNAc level and increased TGF-beta1 and p21 expression were found in MCGT1 cells cultured in normal glucose concentration. Rhein treatment decreased all these features of MCGT1 cells but did not exert a direct effect on GFAT enzymatic activity. CONCLUSIONS AND IMPLICATIONS: There was over-activity of the hexosamine pathway in MCGT1 cells, which may explain the higher expression of TGF-beta1 and p21, the cellular hypertrophy and the increased expression of extracellular matrix (ECM) components in the cells. By inhibiting the increased activity the hexosamine pathway, rhein decreased TGF-beta1 and p21 expression and thus contributed to the decreased cellular hypertrophy and ECM synthesis. Inhibition of the hexosamine pathway may be one of the mechanism through which rhein exerts its therapeutic role in diabetic nephropathy.


Subject(s)
Anthraquinones/pharmacology , Diabetic Nephropathies/drug therapy , Enzyme Inhibitors/pharmacology , Glucose Transporter Type 1/drug effects , Animals , Cells, Cultured , Cyclin-Dependent Kinase Inhibitor p21/drug effects , Cyclin-Dependent Kinase Inhibitor p21/metabolism , Diabetic Nephropathies/physiopathology , Extracellular Matrix/drug effects , Extracellular Matrix/metabolism , Gene Expression Regulation/drug effects , Glucose Transporter Type 1/metabolism , Hexosamines/metabolism , Hypertrophy/metabolism , Male , Mesangial Cells/drug effects , Mesangial Cells/metabolism , Phenotype , Rats , Rats, Sprague-Dawley , Transforming Growth Factor beta1/drug effects , Transforming Growth Factor beta1/metabolism
12.
Exp Clin Endocrinol Diabetes ; 115(9): 571-6, 2007 Oct.
Article in English | MEDLINE | ID: mdl-17943690

ABSTRACT

OBJECTIVE: Rhein and angiotensin-converting enzyme inhibitor (ACEI) have been reported to prevent the progression of diabetic nephropathy (DN). We further explore the unknown ability to induce renal-protection of rhein and ACEI combined therapy in DN compared with the therapeutic effects of single treatment of them by using db/db mouse of type 2 diabetes model. METHODS: db/db and db/m mice, 8 weeks of age, were divided into five groups according to the following treatments: (A) db/m, given saline treatment; (B) db/db, given saline treatment; (C) db/db, given rhein treatment (150 mg/kg/day); (D) db/db, given benazepril treatment (10 mg/kg/day); (E) db/db, given rhein (150 mg/kg/day) with benazepril (10 mg/kg/day). Body weight, plasma glucose, plasma lipid and 24 h urinary albumin excretion levels were measured every 4 weeks. Morphometry of renal tissue and immunohistology of transforming growth factor-beta1 (TGF-beta) and fibronectin were determined for all groups at the end of the treatment. RESULTS: It was found that after treatment urinary albumin excretion was reduced after 4 weeks treatment in group E and after 8 weeks treatment in groups C and D, when compared to group B (p<0.05). Plasma creatinine levels dropped significantly for group E, compared with the diabetic control group by the end of the treatment period. Furthermore, after the treatment body weight, plasma glucose, cholesterol, triglyceride and low density lipoprotein all decreased in groups C and E compared to group B (p<0.05). Histological morphometric analysis revealed that the whole glomerular area and extracellular matrix area was significantly reduced in groups C, D and E compared to group B, at 20 weeks of age, an effect most pronounced in group E. Using immunohistochemistry, the expression of fibronectin and TGF-beta1 in groups C, D and E was found to have decreased compared to group B, after 12 weeks treatment, again the effect being more pronounced in group E. CONCLUSIONS: There appeared to be a similar renal protective effect of rhein compared with benazepril in diabetic nephropathy. A combined therapy may offer a more beneficial complementary effect on kidney injury in db/db mice, as reflected by urinary albumin excretion, renal function and histological changes. Our findings suggest that a therapeutic approach that combines rhein with ACEI provides a more effective therapy for DN than does either agent alone.


Subject(s)
Angiotensin-Converting Enzyme Inhibitors/pharmacology , Anthraquinones/pharmacology , Benzazepines/pharmacology , Diabetes Mellitus, Type 2/drug therapy , Diabetic Nephropathies/drug therapy , Albuminuria/drug therapy , Animals , Blood Glucose/metabolism , Cholesterol/blood , Diabetes Mellitus, Type 2/blood , Diabetes Mellitus, Type 2/urine , Diabetic Nephropathies/blood , Diabetic Nephropathies/prevention & control , Diabetic Nephropathies/urine , Drug Therapy, Combination , Enzyme Inhibitors/pharmacology , Fibronectins/metabolism , Fluorescent Antibody Technique, Direct , Immunohistochemistry , Mice , Mice, Inbred C57BL , Random Allocation , Transforming Growth Factor beta/metabolism , Triglycerides/blood
13.
Histopathology ; 51(2): 195-203, 2007 Aug.
Article in English | MEDLINE | ID: mdl-17573940

ABSTRACT

AIMS: To evaluate primary and recurrent embryonal sarcoma of the liver and to improve recognition of its morphological variants and immunohistochemical features. METHODS AND RESULTS: Fourteen primary and two recurrent cases of hepatic embryonal sarcoma were evaluated histologically and investigated immunohistochemically with a panel of antibodies using the EnVision+ system. They were usually single, large, globular masses with solid and cystic gelatinous areas. Microscopic features included spindle, oval, stellate, epithelioid or multinucleated cells loosely or densely arranged in a myxomatous matrix. Entrapped bile ducts and hepatic cords were often present at the periphery of the tumours. Intracellular and extracellular periodic acid-Schiff-positive, diastase-resistant hyaline globules were commonly present. Recurrent tumours showed greater cellularity, anaplasia and pluripotential differentiation compared with the primary tumour. Immunohistochemistry showed evidence of widely divergent differentiation into mesenchymal and epithelial phenotypes. CONCLUSIONS: Embryonal sarcoma of the liver may undergo pluripotential differentiation and diagnosis should be based mainly on morphological features. Immunohistochemistry has no specific or diagnostic relevance, but, by using a panel of antibodies, may help to exclude other tumours.


Subject(s)
Liver Neoplasms/metabolism , Liver Neoplasms/pathology , Neoplasms, Germ Cell and Embryonal/metabolism , Neoplasms, Germ Cell and Embryonal/pathology , Actins/metabolism , Adolescent , Adult , Biomarkers, Tumor/metabolism , Cell Differentiation , Child , Diagnosis, Differential , Female , Humans , Immunohistochemistry , Male , Middle Aged , Neoplasm Recurrence, Local/metabolism , Neoplasm Recurrence, Local/pathology , Proteins/metabolism , alpha 1-Antitrypsin/metabolism , alpha-Fetoproteins
14.
Pharmazie ; 57(7): 482-4, 2002 Jul.
Article in English | MEDLINE | ID: mdl-12168531

ABSTRACT

The purpose of this study was to investigate electroporation and iontophoresis as a means for in vitro delivery of Defibrase--a thrombin-like enzyme (TLE) from Agkistrodon halys ussuriensis Emelianov snake venom--through human epidermis membrane (HEM). Electroporation was carried out using an exponential decay pulse generator (BioR-ad Genepulser, USA) for a period of 0.5 h, followed by a period of 5.5 h passive diffusion or iontophoresis. The results indicated that the combined use of electroporation and anodal iontophoresis in pH 6.4 permeation medium could effectively enhance the skin permeation of Defibrase, whose apparent permeability coefficient was 1.6 +/- 0.8 x 10(-4) cm.h-1. The delivery of Defibrase by the combined use of electroporation and anodal iontophoresis was more effective than by electroporation alone (P < 0.01) or by the combined use of electroporation and cathodal iontophoresis (P < 0.01). Moreover, when the pH of the permeation medium was raised from 6.4 to 7.4 the permeation of Defibrase caused by a combined use of electroporation and anodal iontophoresis showed a tendency to increase. These results implied that electroosmotic flow effect might be important for the iontophoretic (following electroporation) skin permeation of Defibrase.


Subject(s)
Batroxobin/pharmacokinetics , Crotalid Venoms/pharmacokinetics , Fibrinolytic Agents/pharmacokinetics , Skin Absorption/physiology , Electric Stimulation , Electrodes , Hydrogen-Ion Concentration , Iontophoresis
15.
Steroids ; 66(12): 905-10, 2001 Dec.
Article in English | MEDLINE | ID: mdl-11711119

ABSTRACT

OBJECTIVE: Estrogen-dependent growth of breast cancer can be blocked by anti-estrogens. Estrogen receptor (ER) presence in breast cancer implies responsiveness to endocrine therapy. However, for those patients who ultimately develop resistance to endocrine therapy, the mechanisms remain unclear. The present study attempted to compare the expression status of ER mRNA in a series of primary breast tumors with matched metastases and explored the relation between ER and mutant p53 expression. METHODS: In situ hybridization using a digoxigenin-labeled estrogen receptor cDNA probe was employed to determine the expression of ER mRNA in 52 cases of primary tumors and their matched axillary lymph node metastases. Immunohistochemical staining using a monoclonal antibody against ER was also performed. RESULTS: ER expression was observed in 53.8% (28/52) of primary tumors and 48% (25/52) of metastases, while 57.7% (30/52) of primary tumors and 53.8% (28/52) of metastases showed ER mRNA positivity. There were variations in ER status between in situ hybridization and immunohistochemistry measurements and between primary tumors and metastases. Mutant p53 expression was inversely associated with ER-negative, high-grade tumors. CONCLUSIONS: In situ hybridization may be a more specific and sensitive method for determination of ER status than immunohistochemistry. It is possible that the biologic properties of ER change, and these changes may influence tumor response to endocrine therapy. In view of the ER variation, it was suggested that the ER status of metastatic tumors in addition to primary tumors should be taken into consideration in order to better determine the benefit of clinical endocrine therapy.


Subject(s)
Breast Neoplasms/genetics , Breast Neoplasms/pathology , Gene Expression Regulation, Neoplastic , Genes, p53/genetics , Lymphatic Metastasis/genetics , Receptors, Estrogen/genetics , Female , Genetic Predisposition to Disease , Humans , Immunohistochemistry , In Situ Hybridization , Middle Aged , Mutation/genetics , RNA, Messenger/genetics , RNA, Messenger/metabolism , Receptors, Estrogen/metabolism
16.
Shi Yan Sheng Wu Xue Bao ; 34(4): 279-82, 2001 Dec.
Article in Chinese | MEDLINE | ID: mdl-12549206

ABSTRACT

Exchange plasmid pF-HPRT-F3 and Flp expression plasmid pCMV-Flp were constructed and then introduced using electroporation system into F18 ES cell line, which have an exchange cassette F-Neo-F3 at HPRT locus. After HAT selection, HAT resistant clones were obtained. Then G418 sensitivity test and Southern blotting were carried out to screen RMCE recombinants. The results indicated that RMCE had taken place in three of 12 HAT resistant clones. The frequency is 25%. The result demonstrates that it is realizable to introduce transgene to HPRT locus by using Flp recombinase mediated cassette exchange reaction.


Subject(s)
DNA Nucleotidyltransferases/physiology , Hypoxanthine Phosphoribosyltransferase/genetics , Animals , Cell Line , Clone Cells , Electroporation , Genetic Vectors , Mice , Mutagenesis, Insertional , Plasmids/genetics , Transfection
17.
Yao Xue Xue Bao ; 36(9): 707-10, 2001 Sep.
Article in Chinese | MEDLINE | ID: mdl-12580114

ABSTRACT

AIM: To study the effect of pH on the transcorneal permeability of timolol maleate (TM). METHODS: The apparent distribution coefficients of TM were determined. The permeability of TM across isolated rabbit cornea was measured using in vitro method at various pH. RESULTS: The partition coefficient and pKa of TM were 63.63 and 9.17 respectively. At neutral pH, the apparent permeation coefficient of TM was 1.43 x 10(-5) cm.s-1. When the pH varied from 6.65 to 9.20, the cumulative amount for timolol transcorneal penetration increased 1.3 times and the lag time decreased more than 19-folds. The calculated permeability coefficients of ionized and un-ionized timolol were 1.29 x 10(-5) cm.s-1 and 4.22 x 10(-5) cm.s-1, respectively. CONCLUSION: Timolol penetrated corneal membrane mainly as free base by intracellular pathway, and corneal epithelium was the rate-limiting barrier.


Subject(s)
Antihypertensive Agents/pharmacokinetics , Cornea/metabolism , Timolol/pharmacokinetics , Animals , Epithelium, Corneal/metabolism , Female , Hydrogen-Ion Concentration , In Vitro Techniques , Male , Permeability , Rabbits
18.
Sheng Wu Gong Cheng Xue Bao ; 17(5): 566-9, 2001 Sep.
Article in Chinese | MEDLINE | ID: mdl-11797223

ABSTRACT

Using the HPRT genomic DNA fragment and synthesized oligonucliotides, pSP-HPRT-F-Neo-F3 was designed and constructed as a replacement gene targeting vector by usual molecular cloning techniques. Structure of pSP-HPRT-F-Neo-F3 was identified by restrictive digestion analysis and partly sequencing. Then linearized pSP-HPRT-F-Neo-F3 DNA was electroporated into ES cells, and transfected cells were screened by being cultured in medium containing 200 micrograms/mL G418 and 2 micrograms/mL 6-GT. Twenty-four double drug resistant clones were picked up and analyzed, among them, two clones were proved to have taken place the required recombination by PCR and southern blotting analysis.


Subject(s)
DNA Nucleotidyltransferases/genetics , Embryo, Mammalian/metabolism , Gene Targeting/methods , Animals , Blotting, Southern , Cell Line , Clone Cells , DNA/genetics , DNA Nucleotidyltransferases/metabolism , DNA, Recombinant/genetics , Embryo, Mammalian/cytology , Genetic Vectors/genetics , Hypoxanthine Phosphoribosyltransferase/genetics , Hypoxanthine Phosphoribosyltransferase/metabolism , Mice , Polymerase Chain Reaction , Transfection
19.
Acta Crystallogr D Biol Crystallogr ; 56(Pt 7): 918-9, 2000 Jul.
Article in English | MEDLINE | ID: mdl-10930844

ABSTRACT

Mabinlin II is a sweet protein with the highest known thermostablility and is isolated from the seeds of Capparis masaikai Levl. grown in south China. Two crystal forms of mabinlin II were obtained using the hanging-drop vapour-diffusion method. One of them diffracts to 2.8 A resolution and belongs to space group P2, with unit-cell parameters a = 50.16, b = 50.17, c = 76.60 A, beta = 99.6 degrees. There are four molecules per asymmetric unit, with a solvent content of 35.3%.


Subject(s)
Plant Proteins/chemistry , Sweetening Agents/chemistry , Crystallization , Crystallography, X-Ray , Molecular Structure , Temperature
20.
Yao Xue Xue Bao ; 32(10): 777-81, 1997 Oct.
Article in Chinese | MEDLINE | ID: mdl-11596223

ABSTRACT

In this paper, poloxamer and PVP were used as carriers. CyA-poloxamer and CyA-PVP solid dispersions were prepared using the melting-solvent method or solvent method, respectively. The percentage compositions of the solid dispersions ranged from 10:90 to 90:10. Hot stage microscopy, powder X-ray diffraction and differential scanning calorimetry were used to examine the physical-chemical characteristics of the solid dispersions to provide some reliable and scientific basis for preformulation of CyA.


Subject(s)
Cyclosporine/chemistry , Calorimetry, Differential Scanning , X-Ray Diffraction
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