To examine whether or not the regulatory sequence of chicken ovalbumin gene can drive transgene expression specifically in hen oviduct, the authors constructed an oviduct-specific expression vector (pOV), containing 3.0 kilobases (kb) of the 5'-flanking sequence and 3.0 kb of the 3'-flanking sequence of the chicken ovalbumin gene. Jellyfish green fluorescence protein (EGFP) reporter gene and bacterial LacZ reporter gene were respectively inserted into the downstream of the 5'-regulatory region.The recombinants were named as pOVEGFP and pOVLacZ. Two transfer systems, in vitro and in vivo, were used to verify the function of the vector. In vitro, the plasmid DNA pOVEGFP and pEGFP-N1 were transfected respectively by the polyethyleneimine procedure into the primary chicken oviduct epithelium (PCOE) and fibroblasts cells isolated from laying hens. In vivo, the recombinant vector pOVLacZ was injected into egg-laying hens via wing vein and the tissues were collected for RT-PCR analysis.The results showed that expression of pEGFP-N1 was achieved at low level in oviduct epithelial cells and at high level in fibroblasts, but that the recombinant vector was not expressed in both cells. RT-PCR analysis showed that the LacZ gene was transcribed in the oviduct, but not in the heart, liver, kidney and spleen of the injected hens. Accordingly, the ?-galactosidase activity was only detected in the oviduct magnum (116.7 mU/ml) and eggs (16.47 mU/ml). These results indicated that the cloned regulation regions of chicken ovalbumin gene could drive exogenous gene expression specifically in the oviducts of hens. In vivo gene injection via wing vein may serve as a rapid production system of recombinant proteins in chicken eggs. In addition, the cultured primary oviduct cells from laying hens were not efficient temporary expression systems for analyzing the function of regulating elements of ovalbumin gene.

Objective To compare proprotein convertase subtilisin/kexin type 9 (PCSK9) levels between premenopausal and postmenopausal women,and to investigate the relationship between serum PCSK9 and metabolic factors.Methods Totally 515 women were enrolled from the study on diabetes of prediction,prevention,and intervention in Nanjing in 2009.Survey,physical examinations,and determination of related metabolic indexes were performed.Serum PCSK9 level was measured by sandwich ELISA.Results Serum PCSK9 level was positively correlated with low density lipoprotein-cholesterol (LDL-C),total cholesterol (TC),triglyceride,fasting plasma glucose,body mass index,waist-hip ratio,and age in women (all P＜0.01).PCSK9 level was significantly lower in premenopausal women than that in postmenopausal women [(58.18 ± 25.44 vs 80.91 ± 33.74) ng/ml,P ＜0.01].Conclusion Higher level of PCSK9 exists in postmenopausal women compared with premenopausal women.The level of PCSK9 is closely correlated with age,TC,and LDL-C.

Aged ; Humans ; Male ; Neoplasms, Muscle Tissue ; Skull Neoplasms ; Temporal Bone ; pathology

Objective To evaluate the value of reconstruction of three-dimentional CT in development dislocation of hip(DDH)in children.Methods Twelve cases of DDH concluded 4 bilateral and 8 unilateral cases.To sum up,16 sick hips were operated and 8 normal hips were also obtained by three-dimensional CT(Hip speed Fi/x,GE Co).Results 3D reconstruction were used to show femoral head,(acetabulum) and relationship of acetabulum and femoral head respectively.The difference between FNA measurement of sick hips and those of normal hips were significant(P

Objective To investigate the clinical epidemiologic features of transient synovitis(TS) of hip in children occurred in Shen-zhen district.Methods The medical files were reviewed and a standard questionnaire was filled according to the conditions of 705 cases such as pathogeny,clinical manifestation,therapy and prognosis.Results Transient synovitis occurred in a sporadic form all the year round.The peak age of patients with TS was 3-7 years old.The ratio of boys to girls was 2.9:1.About 19.3% patients were attacked an upper respiratory tract infection and 11.9% patients attributed the symptoms to trauma or severe activities before 1 week.A varying degree of painful limp and restriction of movement at the hip were found clinically.All of cases were cured by skin traction.The incidence of recurrence was 6.95%.Conclusions Male predominance is found in TS.It is characteristic of sporadic form in the 4 seasons and intently relation to an upper respiratory tract infection and trauma or severe activities.TS is recurrent and the prognosis is good by skin traction.

To examine whether or not the regulatory sequence of chicken ovalbumin gene can drive transgene expression specifically in hen oviduct, the authors constructed an oviduct-specific expression vector (pOV), containing 3.0 kilobases (kb) of the 5'-flanking sequence and 3.0 kb of the 3'-flanking sequence of the chicken ovalbumin gene. Jellyfish green fluorescence protein (EGFP) reporter gene and bacterial LacZ reporter gene were respectively inserted into the downstream of the 5'-regulatory region. The recombinants were named as pOVEGFP and pOVLacZ. Two transfer systems, in vitro and in vivo, were used to verify the function of the vector. In vitro, the plasmid DNA pOVEGFP and pEGFP-N1 were transfected respectively by the polyethyleneimine procedure into the primary chicken oviduct epithelium (PCOE) and fibroblasts cells isolated from laying hens. In vivo, the recombinant vector pOVLacZ was injected into egg-laying hens via wing vein and the tissues were collected for RT-PCR analysis. The results showed that expression of pEGFP-N1 was achieved at low level in oviduct epithelial cells and at high level in fibroblasts, but that the recombinant vector was not expressed in both cells. RT-PCR analysis showed that the LacZ gene was transcribed in the oviduct, but not in the heart, liver, kidney and spleen of the injected hens. Accordingly, the beta-galactosidase activity was only detected in the oviduct magnum (116.7 mU/ml) and eggs (16.47 mU/ml). These results indicated that the cloned regulation regions of chicken ovalbumin gene could drive exogenous gene expression specifically in the oviducts of hens. In vivo gene injection via wing vein may serve as a rapid production system of recombinant proteins in chicken eggs. In addition, the cultured primary oviduct cells from laying hens were not efficient temporary expression systems for analyzing the function of regulating elements of ovalbumin gene.
Animals ; Cells, Cultured ; Chickens ; Cloning, Molecular ; methods ; Fallopian Tubes ; metabolism ; Female ; Organ Specificity ; Ovalbumin ; biosynthesis ; genetics ; Protein Engineering ; methods ; Recombinant Proteins ; biosynthesis ; Transfection ; methods ; Women

Associating liver partition and portal vein ligation for staged hepatectomy (ALPPS) could induce extensive and rapid future liver remnant hypertrophy. However, the morbidity for ALPPS is very high. This paper reports a modified ALPPS (associating microwave ablation and portal vein ligation for staged hepatectomy, AMAPS), which was successfully applied in the treatment of huge hepatocellular carcinoma with cirrhosis, and the procedure of operation was greatly simplified. Hence, AMAPS is feasible and safe in selected patients with primary hepatocellular carcinoma and cirrhosis.
Carcinoma, Hepatocellular* ; Fibrosis* ; Hepatectomy* ; Humans ; Hypertrophy ; Ligation* ; Liver ; Liver Cirrhosis ; Microwaves* ; Portal Vein*

Antioxidant activity of bronchial epithelial cells (BECs) plays an essential role in preventing the airway epithelium integrity from damage in structure and function. Integrin expressed by BECs is the receptor of extracellular matrix such as fibronectin (Fn), and it is involved in modulation of proliferation, differentiation and metabolism of the cells. In order to test the hypothesis that integrin-ligand binding reaction supports the ability of cells to withstand oxidant attack, the present study evaluated the antioxidant activity of primary cultured rabbit BECs treated with fibronectin or its sequence Arg-Gly-Asp (RGD peptide), by determining changes in the activity of glutathione peroxidase (GSH-Px), superoxide dismutase (SOD) and catalase (CAT) and in the level of glutathione (GSH). The results are as follows: (1) Fn (10 μg/ml) increased significantly the activity unit of GSH-Px (P<0.05, n=5), which was inhibited by calmodulin-inhibitor W7 　(10－5 mol/L) (P<0.05). Both Fn (5～20 μg/ml) and RGD (15～60 μg/ml) showed a dose-dependent upregulatory effect (respectively r=0.93 and r=0.73). (2) Treatment with Fn increased SOD activity (P<0.01,　n=7), which was abolished by W7 　(P<0.01). (3) Catalase activity was also stimulated by Fn (P<0.05, n=6) and reversed by W7 　(P<0.01). (4) A dose-dependent increase of GSH level was observed in both Fn (r=0.82) and RGD treatment (r=0.84). The data suggest that the binding of integrin with extracellular matrix can upregulate activity of antioxidant enzymes, and increase the content of GSH and improve the ability of BECs to resist oxidant injury.

Objective Clinical data of 19 Chinese patients with 21 hydroxylase deficiency (21OHD) were analyzed to improve the diagnosis and treatment level. Methods Clinical features and laboratory data were collected from 19 patients with 21OHD before and after treatment. Results In male patients, the average age of early appearance of secondary sexual character was (9.3?2.8)yrs, and excess androgen resulted in phallic enlargement. Primary amenorrhea was the most common complaint in female(87.5%), and the signs included a varying degree of labioscrotal fusion and clitoral enlargement. The average level of 17-hydroxy progesterone(17OHP) was (63.42?35.07) ?g/L, and adrenocorticotrophic hormone(ACTH), dehydroepiandrosterone(sodium) sulfate(DHEAS) and testosterone(T) were obviously elevated. CT scan showed bilateral adrenal hyperplasia. The level of 17OHP was significantly decreased after treatment[(63.42?35.07) ?g/L vs (3.15?2.71) ?g/L](P

OBJECTIVETo investigate the effect of adenovirus-mediated endostatin gene transfer on transplanted lung cancer in mice and its mechanism of action.

METHODSTransplant tumor model was induced by subcutaneous inoculation of 2 x 10(6) Lewis lung cancer (LLC) cells into the back of C57BL/6 mice. The mice were treated by intratumoral injection of 2 x 10(9) pfu Ad-mEndostatin. The expression of endostatin in situ and its maintaining time were detected by immunohistochemistry and Western Blot, respectively. The endostatin level in serum was determined by ELISA . The inhibition of tumor growth and changes of survival were recorded and the microvessel density (MVD) was determined by histochemical stainingwith CD31 and CD105 antibodies. The tumor apoptosis was observed by electron microscopy.

RESULTSIn comparison with controls, intratumoral injection of Ad-mEndostatin significantly inhibited the tumor growth and metastasis, and prolonged the survival rate of mice (P < 0.05). Strong positive expression of mEndostatin was seen in the tumor tissue after injection of Ad-mEndostatin, immunhistochemically ostained by mouse endostatin monoclonal antibody, while the control groups showed only very low expression or absence. Serum endostatin concentration was 1540 +/- 560 ng/ml at the second week of administration, the expression of endostatin diminished a month later. The microvessel density (MVD)) decreased from 42.4 +/- 4.8 to 10.5 +/- 3.2 per x 200 magnificetion microscopic field by CD10 staining and from 68.5 +/- 4.5 to 37.5 +/- 4.6 by CD31 staining, respectively (P < 0.05). More apoptotic tumor cells were seen under the transmission electron microscope.

CONCLUSIONEndostatin gene therapy mediated by adenoviral vector efficiently induces expression of endostatin in vivo, and inhibits the growth and metastasis of tumor. It is concluded that its action is targeted to tumor neovasculature and the mechanism is inhibition of tumor angiogenesis.

Adenoviridae ; genetics ; Angiogenesis Inhibitors ; genetics ; metabolism ; therapeutic use ; Animals ; Carcinoma, Lewis Lung ; metabolism ; pathology ; therapy ; Cell Line, Tumor ; Endostatins ; genetics ; metabolism ; therapeutic use ; Genetic Therapy ; Genetic Vectors ; Male ; Mice ; Mice, Inbred C57BL ; Microvessels ; pathology ; Neoplasm Transplantation ; Neovascularization, Pathologic ; pathology ; Random Allocation ; Transfection ; Tumor Burden