1.Expression,Purification of Fusion Protein TGF?-PE40 and the Cytotoxicity of TGF?-PE40 on Tumor Cells
Chinese Journal of Cancer Biotherapy 1996;0(04):-
Objective: To express and purify transforming growth factor ?(TGF?)-pseudomonas exotoxin 40 and investigate its cytotoxic effect on cancer cells overexpressing epidermal growth factor (EGF) receptor. Methods: Recombi nant plasmid pV28 was constructed by inserting the gene coding TGF?-PE40 into the vector pET28a Expression of fusion protein was conducted using the host BL21. Production of the recombinant protein was induced by IPTG, following extraction and purification of inclusion bodies with His-tag purification system. Cell viability assay (by MTT) was performed to determine the cytotoxic effect of TGF?-PE40 on cancer cells ( A431 and SK-OV3). Results: Recombinant plasmid pV28, which expresses TGF?-PE40, was constructed successfully. Purity of TGF?-PE40 was about 98% after a purification procedure using His-tag column. Cytotoxic experiment showed that at a concentration of 0. 86?0. 07 UUUUg/ml, TGF?-PE40 could reduce 50% viability of A431, which has high expression of EGFR. Whereas the IC50 for ovarian cancer cell SK OV3, which expresses less EGFR, was 6.37?2.18 ?g/ml. There was a significant difference between these two groups (P
2.Effects of sodium hyaluronate on the expression of TGF-β3 and Smad 6 of the epidural scar tissue
Yueliang ZHENG ; Zhimin XIA ; Hui ZHOU ; Wanjun ZHAO
Chinese Journal of Emergency Medicine 2008;17(9):956-960
Objective To study the effect of sodium hyaluronate on the expression of transforming growth factor-β3(TGF-β3) and Smad 6 of the epidural scar tissue. Method Totally 60 male SD rats were randomly divided into two groups:the control group(A, n = 30), sodium hyaluronate group(B, n = 30). 0.25 × 1 cm2 dura mater uncovered area laminectomy was performed at L4 and L5, covered with 0.3 ml sodium hyaluronate in group B, covered with same amount of saline in group A. The animals were sacrificed at 2, 4 and 8 weeks after operation. The specimens were prepared for determination of the expression of TGF-β3 and Smad 6 at scar tissue and the degree of scar adhesion according to Rydell method, and observed the ultrastructure changes of scar tissue with transmission electron microscope. Results At 2 weeks after operation, the expression of TGF-β3 mRNA of two groups were 0.22 and 0.257 ( P = 0.027), respectively. At 4 weeks, group B was was increased significantly, and the mean numbers were 0.362 and 0.411 (P = 0.006). At 8 weeks, the expression of TGF-β3 mRNA of group A was increased significantly, too, but the difference between two groups was significantly, they were 0.427 and 0.470 (P =0.015), respectively. The trend of the expression of TGF-β3 and Smad 6 mRNA was similar. At 2, 4, 8, weeks, the expression was 0.169 and 0.205 (P = 0.089), 0.294 and 0.351 (P = 0.031), and 0.469 and 0.543 (P = 0.021), respectively.In group B the duramater adhesion was decreased (P < 0.05), the proliferation of fibroblasts and fihroblastic function were inhibited (P < 0.05). Conclusions Sodium hyaluronate up-regulated the expression of TGF-β3 and Smad6 and reduced the proliferation and collagen synthesis in fibroblast culture in the scar tissue.
3.The clinical and imaging characteristics of sellar regionlesions in children
Lihong LI ; Yuhua LI ; Hui ZHENG ; Wenjun CAO ; Zhengrong XIA
Journal of Practical Radiology 2017;33(4):593-596,652
Objective To evaluate the clinical and imaging features of the sellar region lesions in children.Methods The clinical and imaging features of 112 cases with sellar region lesions were analyzed retrospectively,which were confirmed by pathology.Results 37 cases were craniopharyngiomas, which had two main symptoms of intracranial hypertension(57%)and diabetes insipidus(11%).On imaging it demonstrated as a calcified cystic tumor(81%).18 cases were gliomas, the main clinical feature of which was decreased visual acuity,13 of them were pilocytic astrocytoma, which manifested as a solid tumor with significantly enhancement(94%).16 cases were germ cell tumors,the main complaint was diabetes insipidus (75%),13 of them were germinoma, which showed iso-high density on CT and moderate enhancement after administration of contrast.And it showed high signal intensity on DWI.7 cases were hamartoma,86% patients of which showed gelasmus epilepsy,the imaging showed iso-signal masses with no enhacement in the hypothalamic papillary region.5 cases were LCH,80% of which had the complaint of diabetes insipidus,imaging findings manifested as thinkened pituitary stalk and loss of hyperintensity of posterior pituitary on T1WI.3 cases were pituitary tumors.23 cases were Rathke's cleft cysts,3 cases were arachnoid cyst.Most of the patients presented with headache.Conclusion The clinical and imaging features shows some specific features, which is helpful to improve the correct rate of diagnosis and provide the basis for further treatment.
4.Survey and thought of laboratory medicine undergraduates
hui, XIA ; mei-fang, ZHENG ; ming-min, GU
Journal of Shanghai Jiaotong University(Medical Science) 2008;0(S1):-
Objective To investigate students' view about the laboratory medicine curriculum and teaching reform,and to provide reference for further reform and development. Methods The students specialized in laboratory medicine in School of Medicine,Shanghai Jiaotong University were involved in the survey.Questionnaires were answered and analyzed.The research method of expert consultation was adopted. Results Students had some different opinions for course content,structure and so on,and they expected more clinical practice,more hands operation,and more teachers' guidance.Conclusion To train laboratory technicians,it was necessary to build a reasonable system of professional courses and train practical talents.
6.Effect of zuoguiyin on the expression of ovarian follicle-stimulating hormone receptor in rats during peri-menopausal period.
Wei ZHAO ; Hai-xia WEN ; Hui-li ZHENG
Chinese Journal of Integrated Traditional and Western Medicine 2010;30(3):286-290
OBJECTIVETo investigate the effect and mechanism of Zuoguiyin (ZGY, a Chinese medical remedy for regulating qi and nourishing yin) on expression of ovarian follicle-stimulating hormone receptor (FSHR) in rats during peri-menopausal period (PMP).
METHODSPMP rats were administered with low (13.78 g/kg), middle (20.67 g/kg) and high (31.00 g/kg) dose ZGY respectively by gastrogavage for eight weeks. FSHR mRNA and protein expressions were detected by RT-PCR and immunohistochemistry respectively. Besides, granulosa cells, collected from pregnant mare serum treated nonage rats, were incubated, and the level of FSHR mRNA expression in the cultured cells were detected after various disposals.
RESULTS(1) Ovarian levels of FSHR mRNA and protein expressions in PMP rats were significantly lower than those in young rats (P<0.01), but were up-regulated significantly by ZGY treatment (P<0.01). (2) As compared with the control, FSHR mRNA expression increased in cultured granulosa cells after treated by ZGY extract; the increasing effect of ZGY was enhanced by combined treatment with Forskolin and attenuated by Genistein (a tyrosine protein kinase inhibitor).
CONCLUSIONZGY could improve the ovarian functions during PMP by up-regulating the expression of FSHR and raise the ovarian responsibility to FSH, which may be possibly realized by activating cAMP-protein kinase and tyrosine protein kinase signal pathway.
Animals ; Cells, Cultured ; Drugs, Chinese Herbal ; pharmacology ; Female ; Ovary ; drug effects ; metabolism ; Perimenopause ; drug effects ; RNA, Messenger ; genetics ; Rats ; Rats, Wistar ; Receptors, FSH ; metabolism
7.A preliminary clinical study on two kinds of ultrasonic elastographic technique for diagnosis of hepatic fibrosis
Hui FENG ; Xinli ZHANG ; Meng LI ; Yanxian ZHOU ; Min CHEN ; Xia CHEN ; Zheng DONG
Chinese Journal of Ultrasonography 2010;19(3):227-230
Objective To explore the clinical value of virtual touch quantification technique(VTQ)and fibroscan technique for the diagnosis of hepatic fibrosis.Methods A total of 1 02 patients with chronic liver disease and 78 normal individuals were enrolled in the study.They were all examined with VTQ and fibroscan technique.Pathological results were used as standard criterion.Results The liver tissue riqidity was associated with pathological results.The coefficient of relativity was 0.43309(VTQ)and 0.35840(Fibroscan).ROC curve displayed that VTQ value of 1.4 m/s and fibroscan value of 7.75 kPa can be used to differential diagnose the lowgrade liver fibrosis and high-grade liver fibrosis.The probability of success was 100 0A(102/102)and 100%(78/78)by VTQ,but 88%(90/102)and 100%(78/78)by fibroscan technique.Conclusions VTQ and fibroscan technique are useful in the diagnosis of hepatic fibrosis.Compared with fibroscan technique,VTQ has more advantages in sensitivity practicability and convenience.
8.Fasting plasma obestatin levels in patients with different glucose tolerance and its association with lipid metabolism
Pengying GU ; Dongmei KANG ; Yuan XIA ; Hui ZHENG ; Weidong WANG ; Jian ZHOU ; Cuiping ZHAO ; Yan CHEN
Chinese Journal of Geriatrics 2010;29(6):480-482
Objective To investigate whether fasting obestatin level is different in patients with impaired glucose tolerance or type 2 diabetes, and to explore the association between obestatin and lipid metabolism. Methods Eighty-four subjects without known diabetes were divided into three groups: normal glucose tolerance(NGT), impaired glucose tolerance (IGT) and type 2 diabetes (DM) Plasma obestatin levels were measured with a radioimmunoassay. The relationship between fasting obestatin levels and metabolic parameters was also analyzed. Results Fasting obestatin levels were lower in DM group [(2.82±0.78)ng/ml] and IGT group [(3.25±0.29)ng/ml] than in NGT group[(3.55±0.57) ng/ml, P<0.01]. Triglycerides and low density lipoprotein cholesterol levels gradually increased among the three groups (P<0.05). Multiple linear regression analysis revealed fasting obestatin level was independently associated with waist-to-hip ratio, triglyeride and low density lipoprotein cholesterol. The regression equation was obestatin=6.953-3.412×W/H-0.175×TG-0.123×LDL-C. Conclusions The decreased obestatin may be associated with IGR and T2DM, and obestatin level may be associated with lipid metabolism.
9.Hypothesis of 'saturated or nonsaturated cytotoxicity model' clarifies antibodies mediated immunoreaction in transplant recipient
Bicheng CHEN ; Hui GUO ; Yong LIU ; Sheng CHANG ; Yong CAI ; Peng XIA ; Shaoling ZHENG ; Yirong YANG
Chinese Journal of Organ Transplantation 2011;32(6):351-354
Objective To develop the hypothesis ‘saturated or non-saturated cytotoxicity model' and explain the various phenomena of antibody mediated immunoresponses in recipients,including rejection and accommodation.Methods The imitating complement dependent cytotoxicity.The threshold set to identify as saturated or non-saturated cytotoxicity depends on antigen-antibody complex(R)whether or not above lethal number(D)in effective time.Feasibility of the hypothesis was examined through explaining various phenomena mediated by anti-donor antibodies,especially some contradictory phenomena.Results Hyperacute rejection,accelerated rejection and acute rejection could be well explained by saturated cytotoxicity.Accommodation of ABO imcompatible transplantion,de novo antibody induced injury,change of protein profile,and C4d deposition in graft could be well elucidated by the hypothesis.Conclusion The hypothesis saturated or nonsaturated cytotoxicity model' help to interpret and interconnect various phenomena of antibodies mediated immune response,such as rejection and accommodation.
10.Use of the Y-shaped mesh in functional repair of the pelvic floor in women
Yudi ZHANG ; Dan LU ; Ping ZHENG ; Xia WU ; Hui LI ; Juhong LIU
Chinese Journal of Tissue Engineering Research 2016;20(30):4503-4508
BACKGROUND:The Y-shaped mesh graft material weaved using lightweight polypropylen has the appropriate porosity, which not only can make the vaginal tissues grow and fuse rapidly on the mesh, but also can maintain good biological strength to ensure the fixed strength for the presacral suspension. OBJECTIVE:To retrospectively analyze the clinical effect of Y-shaped mesh for the biological function reconstruction of the female pelvic floor. METHODS:Ten female cases of pelvic floor dysfunction were enrol ed, aged 37-73 years, and al were given the treatment of sacral colposuspension under laparoscopy. Then perioperative complications were recorded;patients were fol owed up regularly to record the Pelvic Organ Prolapse Quantification (POP-Q) score at different time points;and the subjective satisfaction was investigated using the Pelvic Floor Impact Questionnaire (PFIQ-7). RESULTS AND CONCLUSION:After at least 6-month fol ow-up, no postoperative pelvic infection, nerve damage and complications appeared, the patients healed wel , and no mesh erosion, infection and other adverse reactions occurred. The POP-Q and PFIQ-7 scores at 1, 3 and 6 months after repair were significantly improved than those before repair (P<0.05). These results suggest that the Y-shaped biological mesh repairing female midpelvic floor dysfunction has good biocompatibility, and can restore the anatomy of the pelvic floor.