2.A study on biological activity of co-expression plasmid of human tissue plasminogen activator and vascular endothelial growth factor 165
Zhongjun WU ; Yi ZHU ; De SHI ; Shusen ZHENG
Chinese Journal of General Surgery 1997;0(04):-
Objective To observe the co-expression plasmid of tissue plasminogen activator (tPA) and vascular endothelia growth factor165 (VEGF165) in vascular endothelial cell (VEC) and to study the effect of the product on the proliferation of VEC and fibrinolysis activity. Methods pBudCE4.1/tPA-VEGF165 was transfected into VECs by using lipofection. The expression of tPA and VEGF165 at mRNA level was detected by RT-PCR and expression at protein level was detected by Western blot. The fibrinolysis activity of VEC culture solution of transfecting tPA and VEGF165 genes were detected by fibrin plate technique. The VEC and VSMC were cultured with VEC culture solution of transforming tPA and VEGF165 genes, the proliferation of VEC and VSMC were evaluated with 3?H-TdR incorporation and flow cytometry (FCM). Results The expression of tPA and VEGF165 in the transfected VECs was detected. The fibrinolysis activity of transfected VEC culture solution was also detected. tPA and VEGF165 products in VECs elevated proliferation of VEC, while there was no effect on the proliferation of VSMC. Conclusion The tPA and VEGF165 eukaryotic co-expression plasmid could express in transfected VECs, and the expression products have biology activity.
3.Amelioration of mitochondrial dysfunction in heart failure through S-sulfhydration of Ca2+/calmodulin-dependent protein kinaseⅡ
WU DAN ; HU QING-XUN ; ZHU DE-QIU ; ZHU YI-ZHUN
Chinese Journal of Pharmacology and Toxicology 2017;31(10):1025-1026
OBJECTIVE To determine the functional role of hydrogen sulfide (H2S) in protecting against mitochondrial dysfunction in heart failure through the inhibition of Ca2 +/calmodulin-dependent protein kinaseⅡ (CaMKⅡ) using wild type and CSE knockout mouse models. METHODS Continuous subcutaneous injection isoprenaline (7.5 mg·kg-1·d-1), once a day for 4 weeks to induce heart failure in Male C57BL/6 (6-8 weeks old) mice and CSE-/- mice. 150 μmol·L-1 H2O2 was used to induce oxidative stress in H9c2 cells. Echocardiograph was used to detect cardiac parameters. H&E stain and Masson stain was to observation histopathological changes. Western blot was used to detect protein expression and activity. The siRNA was used to silence protein expression. HPLC was used to detect H2S level. Biotin assay was used to detect the level of S- sulfhydration protein. RESULTS Treatment with S-propyl-L-cysteine (SPRC) or sodium hydrosulfide (NaHS), modulators of blood H2S levels, attenuated the development of heart failure in animals, reduced lipid peroxidation, and preserved mitochondrial function. The inhibition CaMKⅡ phosphorylation by SPRC and NaHS as demonstrated using both in vivo and in vitro models corresponded with the cardioprotective effects of these compounds. Interestingly, CaMKⅡ activity was found to be elevated in CSE-/- mice as compared to wild type animals and the phosphorylation status of CaMKⅡ appeared to relate to the severity of heart failure. Importantly, in wild type mice SPRC was found to promote S-sulfhydration of CaMKII leading to reduced activity of this protein however, in CSE-/- mice S-sulfhydration was abolished following SPRC treatment. CONCLUSION A novel mechanism depicting a role of S-sulfhydration in the regulation of CaMKⅡ is presented. SPRC mediated S-sulfhydration of CaMKII was found to inhibit CaMKⅡ activity and to preserve cardiovascular homeostasis.
4.Amelioration of mitochondrial dysfunction in heart failure through S-sulfhydration of Ca2+/calmodulin-dependent protein kinaseⅡ
WU DAN ; HU QING-XUN ; ZHU DE-QIU ; ZHU YI-ZHUN
Chinese Journal of Pharmacology and Toxicology 2017;31(10):976-976
OBJECTIVE To determine the functional role of hydrogen sulfide (H2S) in protecting against mitochondrial dysfunction in heart failure through the inhibition of Ca2 +/calmodulin-dependent protein kinaseⅡ (CaMKⅡ) using wild type and CSE knockout mouse models. METHODS Continuous subcutaneous injection isoprenaline (7.5 mg·kg-1 per day), once a day for 4 weeks to induce heart failure in male C57BL/6 (6-8 weeks old) mice and CSE-/- mice. 150 μmol·L-1 H2O2 was used to induce oxidative stress in H9c2 cells. Echocardiograph was used to detect cardiac parameters. H&E stain and Masson stain was to observation histopathological changes. Western blot was used to detect protein expression and activity. The siRNA was used to silence protein expression. HPLC was used to detect H2S level. Biotin assay was used to detect the level of S-sulfhydration protein. RESULTS Treatment with S-propyl-L-cysteine (SPRC) or sodium hydrosulfide (NaHS), modulators of blood H2S levels, attenuated the development of heart failure in animals, reduced lipid peroxidation, and preserved mitochondrial function. The inhibition CaMKⅡ phosphorylation by SPRC and NaHS as demonstrated using both in vivo and in vitro models corresponded with the cardioprotective effects of these compounds. Interestingly, CaMKⅡ activity was found to be elevated in CSE-/- mice as compared to wild type animals and the phosphorylation status of CaMK Ⅱ appeared to relate to the severity of heart failure. Importantly, in wild type mice SPRC was found to promote S-sulfhydration of CaMKⅡ leading to reduced activity of this protein however, in CSE-/- mice S-sulfhydration was abolished following SPRC treatment. CONCLUSION A novel mechanism depicting a role of S-sulfhydration in the regulation of CaMKⅡ is presented. SPRC mediated S-sulfhydration of CaMKⅡ was found to inhibit CaMKⅡ activity and to preserve cardiovascular homeostasis.
5.Comparison of the VISX WaveScan and OPD-ScanⅢwith the subjective refraction
Ran, ZHU ; Ke-Li, LONG ; Xiu-Mei, WU ; Quan-De, LI
International Eye Science 2014;(10):1849-1851
AIM:To compare the refractive errors measured by the VISX WaveScan, OPD - Scan Ⅲ and the subjective refraction.
METHODS: Seventy - six patients ( 152 eyes ) were recruited from January 2013 to December 2013. All patients were measured with subjective refraction by the phoropter (NIDEK, RT-5100), objective refraction by the WaveScan ( AMO Company, USA) , OPD-ScanⅢ ( Nidek Technologies, Japan). The sphere, cylinder, axis of the three methods were compared and analyzed.
RESULTS: The sphere measured by WaveScan was lower than that by subjective refraction, the difference was 0. 13±0. 30D (t=3. 753, P<0. 001). For cylinder, the difference was 0. 13±0. 43D (t=3. 664, P<0. 001). There was no significance for sphere, cylinder, and spherical equivalent between OPD - Scan Ⅲ and subjective refraction (P>0. 05). The value of the difference between WaveScan and subjective refraction was 5. 87o±6. 19o for the axis and the difference between OPD-Scan Ⅲ and subjective refraction was 3. 82o±3. 95o. There was statistic significance (t=2. 817, P=0. 006).
CONCLUSION: For sphere and cylinder, WaveScan generated some deviation relative to subjective refraction. The Nidek OPD-ScanⅢ gives more accurate measures of objective refraction when compared with subjective refraction.
6.Risk factors for lymph node metastasis of early colorectal cancer and indices of endoscopic resection
Ji DE ; Jinyu QIN ; Jin WANG ; Linlin ZHU ; Jinlin YANG ; Yiping WANG ; Junchao WU
Chinese Journal of Digestive Endoscopy 2015;32(6):367-370
Objective To analyse the risk factors for lymph node metastasis of early colorectal cancer and the therapeutic indication for endoscopy.Methods The clinical data of the 269 early colorectal cancer patients who underwent surgical treatments between January 2009 and December 2013 in the West China hospital of Sichuan University were analyzed retrospectively.Pathologic features were compared between different histological types and investigated by univariate and multivariate analysis of their possible risk factors for lymph node metastasis.Results Univariate analysis showed that tumor size (P =0.029),depth of tumor invasion (P =0.006),histological type (P =0.000) and lymphatic involvement (P =0.035) were correlated with lymph node metastasis.Multivariate analysis revealed that tumor size (OR =5.385,95% CI:1.156-25.075,P =0.032)and histological type (OR =5.145,95% CI:1.553-17.053,P =0.007) were independent risk factorsfor lymph node metastasis.Comprehensive analysis showed that lymph node metastasis could not be found in patients with tumor invading the mucous layer.However,lymph node metastasis occurred if the tumor invaded the submucous layer.The larger and less differentiated tumor was,the higher occurrence of lymph node matastasis would be (P < 0.05).Conclusion Endoscopic resection is recommended for those with early colorectal cancer localized in the mucous layer and without lymph node metastasis.It is recommended that patients with submucosal carcinomas undergo a preoperative endoscopic ultrasonography or pathological screening for the most appropriate surgical treatment,as the larger and the lower differentiated the tumor is,the higher chance the lymphatic metastasis is.
7.Influence of pregnant rats' prenatal chronic stress on hippocampal dendritic spines and the function of learning and memory in their offsprings
Xicheng TAO ; De WU ; Jiulai TANG ; Ziyun ZHOU ; Jing ZHU ; Wencheng XU
Chinese Journal of Applied Clinical Pediatrics 2015;30(12):909-912
Objective To study the influence of pregnant rats' prenatal chronic stress (PS) on learning and memory of their offspring rats and its possible molecular mechanisms.Methods Pregnant females were individually restrained for 45 min 3 times a day during pregnancy from day 14 to day 21.Control pregnant females were left undisturbed in their home cages.The rat offsprings were randomly assigned to PS group or control group.Males and females were kept for the study separately.The learning and memory of the developing rat offspring in the Morris water maze were examined.The basal levels of corticosterone (COR) and adreno-cortico-tropic-hormone (ACTH) were analyzed by using radioimmunoassay.The Golgi-Cox impregnation technique was used to compare density and morphology of the CA1 hippocampal dendritic spines.Results The escape latency (EL) to find the platform in the control group was significantly less than that in the PS group in female rat offspring (F =4.533,P < 0.05),and the difference was statistically significant on the 5th day (t =2.788,P < 0.01).EL to find the platform in the control group was significantly less than that in the PS group in male rat offspring (F =6.101,P <0.05),and the difference was statistically significant on the second day (t =3.051,P < 0.01).In the space exploration experiments of the water maze,the retention time observed for the control group and the PS group in the goal quadrant was similar(P > 0.05).The basal levels of the serum COR in the PS group were higher than those in the control group of female rat offspring(t =3.658,P < 0.01) and the basal levels of the serum ACTH in the PS group were higher than those in the control group of male rat offsprings(t =2.319,P < 0.05).A simplified pattern was observed in the CA1 hippocampal dendritic spines in the PS group,showing a less extent of dendritic arborization and the density was significantly lower than that in the control group(t =-3.072,P < 0.01).Conclusions Altered function of the hypothalamic-pituitary-adrenal axis in the offspring mediates the cognitive alterations observed following prenatal stress should to be associated with the lower density and simplified pattern of CA1 dendritic spines.
8.Plasmid-mediated quinolone resistance in clinical isolates of gram-negative bacilli
Xiao-Gang XU ; Shi WU ; Ming-Gui WANG ; Xin-Yu YE ; Yang LIU ; De-Mei ZHU ;
Chinese Journal of Infection and Chemotherapy 2007;0(05):-
Objective To investigate the importance of plasmid-mediated quinolone resistance in the development of quinolone resistance in clinical isolates of gram-negative bacteria.Methods A total of 541 consecutive clinical isolates of gram-negative ba- cilli resistant or intermediate to ciprofloxacin were screened for the qnrA gene by PCR.Conjugation experiments were carried out with azide-resistant E.coli J53 as a recipient.The aac(6')-Ib-cr gene was detected.The mutations in the quinolone-resist- ance-determining region (QRDR) of the gyrA and parC genes were identified in qnrA positive strains.Results qnrA was identi- fied in 7 of the 541 strains.Among the qnrA positive strains,5 were Enterobacter cloacae.No qnrA was detected in nonfer- menters.Quinolone resistance was transferred in 4 of 7 qnrA positive strains.Transconjugants had 12-to 125-fold increases in MIC of ciprofloxacin relative to that of the recipient.Seven strains contained qnrA with a nucleotide sequence identical to that originally reported.Two transconjugants with higher ciprofloxacin MICs contained aac(6')-Ib-cr gene.Mutations occurred in the QRDR of the gyrA and parC genes in 5 PCR-positive clinical strains.Conclusions Transferable plasmid-mediated quinolone resistance associated with qnrA is highly prevalent in clinical strains of Enterobacter spp.aac(6')-Ib-cr gene and mutations in the quinolone targets may co-exist with qnrA,which may contribute to the further increase of resistance to quinolones.
9.Expression of P16,CyclinD1 and P53 in hydatidiform mole and its significance
Xue-Qin WU ; Jin-Quan LIANG ; De-Ju JIANG ; Zheng ZHU ;
Chinese Journal of Primary Medicine and Pharmacy 2006;0(09):-
Objective To study the relationship between P16,CyclinD1 and P53 anti-oncogene and the gen- esis of hydatidiform mole.Methods 30 samples of hydatidiform moles and normal early pregnant aborted placenta villi respectively were obtained to detect the P16,CyclinD1 and P53 anti-oncogene expression in two kinds of tissues by using SP immunohistochemical staining.Results Compared with that of normal villi,the expressions of P16,P53 and CyclinD1 anti-oncogene were quite different in hydatidiform moles.The expression of P16 was all positive,while CyclinD1 and P53 were all negative in the chorion of early gestation.A descending tendency of P16 expression was found,while the expression of CyclinD1 showed an ascending tendency.The positive rate of P16,CyclinD1 and P53 expression was significantly different between the groups.It was also observed that there was significant difference between the P16 and the proliferation trophocyte.Conclusion P16,CyclinD1 and P53 anti-oncogene have a close relationship with the genesis of human hydatidiform mole.
10.Dynamic changes of interleukin-1, interleukin-6 and tumor necrosis factor in i ntermingled skin graft in burned rats
Jianhe WU ; Yanfei ZHAO ; Baoyu HU ; Deyuan LU ; Dean ZHU ; Jixiang SHI
Chinese Journal of Traumatology 2001;4(1):31-36
Objective: To compare the dynamic changes of int erleukin-1 (IL-1), interleukin-6 (IL-6), and tumor necrosis factor (TNF) in intermingled skin graft with those in other types of skin grafts in rats. Methods: A 10%-15% third-degree burn was created in 180 Spreg ue-Dawley (SD) rats. After removing the scar, skin grafts were performed on the open wounds immediately with autoskin (aus, n=54), allosk in (als, n=54) and intermingled skin (n=36). That is to say, in the intermingled skin graft, a big piece of alloskin (mals) was grafted first, and 3 days later, small pieces of autoskin (maus) wer e embedded in the alloskin. The rest 36 rats were taken as the controls. And the biological activities of IL-1, IL-6 and TNF in graft sheets in each group wer e detected after skin graft. Results: The levels of IL-1, IL-6 and TNF in the aus group de creased steadily after their initial elevations, whereas in the als group they i ncreased significantly and kept on the peak level in the later phases. In the in termingled group, there appeared a lowest IL-1 level in the mals and a highest one in the maus simultaneously at 7 (4) days (The number out of parenthesis is t he days after transplanting with alloskin sheets, and the number in parenthesis is the days after embedding autoskin sheets in the intermingled skin graft. Simi larly hereinafter.) after skin graft (P<0.01), and the high level in the maus abruptly decreased at 14 (11) days after skin graft. At exactly the same phase on day 7 (4), a prominent peaked IL-6 in the mals occurr ed. In the later phases, the levels of TNF remained relatively low both in the m als and in the maus. From day 7 (4) on, each cytokine fluctuation in the mals sy nchronized with that in the maus. The longer the post transplantation period las ted, the more the positive cytokine correlated between the mals and the maus. Conclusions: The low levels of IL-1 and TNF may be important f actors to lighten the intensity of local rejection in the intermingled skin graf t. The temporarily peaked IL-6 is both an inducer which induces the production of local IL-1 receptor antagonists and soluble TNF receptors and a signal which indicates a local enhancement of Th2 cells. The mild rejection process and th e synchronized cytokine level during the later phases suggest a possible chimeri sm between the mals and the maus.