1.The Value of CT Perfusion Imaging for Breast Tumor
Jianjian WANG ; Feng CHEN ; Kaihong WU
Journal of Practical Radiology 2001;0(08):-
Objective To research the value of CT perfusion imaging in diagnosing breast tumor .Methods 21 focuses in 17 cases withbreast tumorlike pathological changes diagnosed by molybdenum target examination were scanned by CT perfusion imaging . Cine model,0.5 s/circle ,5 mm in thickness?4,120 kV,60 mA,delayed time 10 s,scan time in all 50 s ,GE 4.2 workstation and perfusion 3 software were used ,the data of creating dynamic picture and the relative informations that image intention changes with time change were analyzed and every parameters related with perfusion were studied and analyzed statistically .Results Blood flow(BF),blood volume(BV),mean transit time(MTT) and permeability surface(PS) were (36.46?17.62)ml?min-1?100 g-1,( 13.76?8.59) ml/100 g,( 28.23?15.75) s and (16.45?12.36) ml?min-1?100 g-1.In cancer group,( 17.35?10.67)ml?min-1?100 g-1,( 4.63?3.47) ml/100 g,( 25.52?12.91) s and ( 3.57?3.36) ml?min-1?100 g-1 in benign lesion group,respectively,there was significant difference between two groups in BF,BV and PS(P
2.Construction and Verification of High-capacity Ribosome Display Single-chain Fv Library
Kaihong HUANG ; Xuexian LI ; Yingting CHEN
Chinese Journal of Practical Internal Medicine 2006;0(18):-
Objective To construct high-capacity ribosome display single-chain Fv library for selection of high affinity ScFv antibody.Methods We isolate human lymphocyte from peripheralblood(2 normal,3 gastric cancer,3 colonic cancer,1 pancreatic cancer,each 5 mL and 2 newborn,each 2 mL)and extract RNA for cloning whole human heavy chain and light chain gene by RT-PCR.VH and VL were rearranged randomly by SOEing(splicing by overlap extension,SOEing).Finally,the elements for in vitro screening such as T7 promoter and ribosome binding site were introduced while the SOEing products were amplified.Moreover,ribosome display template were verified by blue/white screening and further sequencing.Results We successfully constructed ribosome display ScFv library with a volume of 1.1?1013.Conclusion The construction of high-capacity ScFv library shed light on multiple therapeutic ScFv screening.
3.Influencing factors for in-stent restenosis in patients with coronary heart disease after sirolimus-eluting stent implantation
Ganyang LI ; Yong FANG ; Weiguo LI ; Guoyong HUANG ; Kaihong CHEN
Chinese Journal of cardiovascular Rehabilitation Medicine 2015;24(6):610-613
Objective:To study influencing factors for in‐stent restenosis (ISR) during one year in patients with coro‐nary heart disease (CHD) after coronary sirolimus‐eluting stent (SES) implantation .Methods :According to results of coronary angiography (CAG) ,a total of 275 patients ,who hospitalized in our department from Jan 1st ,2012 to Dec 30th ,2013 and have received SES implantation and reviewed CAG after one year ,were divided into non‐ ISR group (n=247) and ISR group (n=38) .Clinical characteristics were compared between two groups ,and Logistic regression analysis was used to analyze influencing factors for ISR .Results:Compared with non‐ISR group ,there were significant rise in percentages of occlusion lesions (17. 9% vs .31. 9% ) ,multiple overlapping stents (16. 7% vs . 31.9% ) ,and significant reduction in percentage of stent post‐dilatation (34.9% vs .10.6% ) in ISR group ,P<0.05 or <0. 01 ;Logistic regression analysis indicated that coronary occlusion lesion was a risk factor (OR :2. 855 ,95%CI :1.197~6.808 ,P=0.018) ,and post‐dilatation was a protective factor (OR :0.198 ,95% CI :0.057~0.691 , P=0.011) for ISR occurrence .Conclusion:Multiple overlapping stents and coronary occlusion lesions increase one‐year in‐stent restenosis rate ;stent post‐dilatation can reduce one‐year in‐stent restenosis rate ;coronary occlusion le‐sions is a risk factor , and stent post‐dilatation is a protective factor for restenosis during one‐year after coronary drug‐eluting stent implantation .
4.Effects of angiotensin II receptor antagonist on remodeling of renal arterioles in hypertension
Xiangqun YANG ; Jianliang ZHANG ; Kaihong JI ; Zonglai JIANG ; Eryu CHEN
Chinese Journal of Pathophysiology 2000;0(08):-
AIM: To investigate the effects of angiotensin II receptor antagonist on remodeling of renal arterioles in hypertension. METHODS: Eighteen 4 weeks old male rats were divided into three groups: Wistar-Kyoto rats (WKY) for normotensive group, and spontaneously hypertensive rats (SHR) for hypertensive group, and SHR treated with losartan orally (15 mg?kg -1 ?d -1 ). The rats were raised to 16 weeks old. The morphometric parameters of the renal arterioles, and the widths of vascular smooth muscle cells (VSMC) and intercellular space were studied on kidney slices by light microscope and electromicroscope respectively, combined with computer-assistant image analysis system. The minimal renal vascular resistance (RVR min ) was studied by isolated kidney perfusion system. RESULTS: The systolic blood pressure of the tail artery, wall thickness, wall area, ratio of wall thickness to inner diameter, width of VSMC of renal arterioles and RVR min were all smaller or lower in losartan group than those of SHR. CONCLUSION: Ang II receptor antagonist losartan can prevent the remodeling of renal arterioles in SHR.
5.Effects of aspirin on cell cycle in pancreatic carcinoma cells
Jiao HAN ; Qikui CHEN ; Zhiqing HUANG ; Kaihong HUANG ; Jingwu CHEN ; Zhaohua ZHU
Chinese Journal of Pathophysiology 1986;0(02):-
AIM: To observe the effects of aspirin and prostaglandin E_2 (PGE_2) on the cell viability and cell cycle in SW1990 human pancreatic carcinoma cell lines, and to investigate the mechanisms of aspirin-induced growth inhibition and cell cycle arrest. METHODS: After incubated with aspirin or PGE_2 and their combination, the viability of SW1990 cells was measured by MTT assay. The levels of intracellular PGE_2 were determined by ELISA. The effects of aspirin or PGE_2 on cell cycle were investigated by flow cytometry (FCM). The expression of p21~ Wafl/cipl and p27~ Kipl/pic2 (the cyclin-dependent kinase inhibitors) were analyzed by Western blotting. RESULTS: Aspirin could inhibit the growth of cells and level of intracellular PGE_2 in a dose-dependent manner. Aspirin enhanced the expression of p21~ Wafl/cipl and p27~ Kipl/pic2 and induced cell cycle arrest at G_0/G_1 phase. PGE_2 increased the cell viability of SW1990 cells. However, it couldn't antagonize the changes of cell viability and cell cycle that induced by aspirin. CONCLUSIONS: The inhibitory effects of aspirin on growth and cell cycle of pancreatic carcinoma cells might not be mediated by a COX-dependent pathway completely. Cell cycle arrest induced by aspirin might be associated with up-regulation of p21~ Wafl/cipl and p27~ Kipl/pic2 .
6.Experimental study of 5-fluorouracil loaded polylactic acid nanoparticles control-releasing preparation on tumor
Kaihong HUANG ; Jianhua LIU ; Lingyun WANG ; Zhaohua ZHU ; Qikui CHEN ; Jun MIN ; Rufu CHEN
Chinese Journal of Pathophysiology 1999;0(09):-
AIM:To investigate the preparation techniques and anti-tumor effects both in vitro and in vivo of a novel nanoparticles control-releasing preparation of 5-fluorouracil(5-FU)by intravenous injection.METHODS:With polylactic acid(PLA)as marix materials,we adopted ultrasound emulsification method to prepare PLA enveloped 5-FU nanoparticles(5-FU-NPs).Scanning electricity microscopy was used to observe the morphology of 5-FU-NPs and laser optical scattering experiment was conducted to determine its diameter distribution.The drug-carrying capacity(ratio)of the nanoparticles was determined by means of high-power liquid chromatography(HPLC)and MTT test was used to observe cytotoxicity in vitro.The anti-tumor effects were determined at different dosages,frequencies of taking drugs in vivo.RESULTS:Scanning electron microscopy showed that the 5-FU-NPs were globular particles with smooth surface in an average particle diameter of 191.9 nm with a normal distribution,and the drug-carrying capacity of 5-FU-NPs was 15.2%.5-FU-NPs had the same anti-cancer effect as unenveloped drug in vitro and showed typical dose-effect relationship.Compared to naked 5-FU,5-FU-NPs presented significant difference(P
7.Nanoparticle for siRNA delivery and its pancreatic cancer targeting abili-ty
Jiajia LI ; Yinting CHEN ; Linjuan ZENG ; Guoda LIAN ; Shaojie CHEN ; Yaqing LI ; Kaihong HUANG
Chinese Journal of Pathophysiology 2014;(9):1567-1573
AIM:To synthesize a safe , efficient and targeted nanoparticulate carrier for siRNA delivery to pan-creatic cancer cells .METHODS: Iron oxide nanocrystal with carboxylic acid group-polyethyleneimine ( IONP-PEI ) was synthesized and investigated as a nonviral carrier of siRNA to the pancreatic cells .The size, surface and charge using zeta potential were characterized .The perfect charge ratio between amino groups of IONP-PEI and phosphate groups of siRNA ( N/P) was determined by the transfection efficiency detection , gel retardation assay and MTS assay .An antibody-directed nonviral vector , scFvCD44v6-IONP-PEI nanoparticle attaching to the cancer-associated CD44v6 single-chain variable frag-ment, was constructed as a cancer-targeting nanocarrier for siRNA delivery .Prussian blue staining and immunofluorescent staining were performed to detect the distribution of scFv CD44v6-IONP-PEI/siRNA complexes in the cells .The transfection efficiency , fluorescence intensity and the expression of KRAS at mRNA and protein levels in the cells transfected by IONP -PEI/siRNA and scFv CD44v6-IONP-PEI/siRNA were detected by flow cytometry , fluorescence microscopy , real-time PCR and Western blotting, respectively.RESULTS:The mass ratio of IONP to PEI was 0.75.The suitable ratio of N/P was 20. The averaged size and surface zeta potential of IONP-PEI/siRNA in deionized water were (51.3 ±2.2)nm (diameter) and (21.73 ±8.07)mV, respectively.Red fluorescence was seen in both targeting and nontargeting groups , which clearly re-vealed the intracellular distribution of siRNA and delivery agents .Transfection efficiencies in targeting and nontargeting groups were (89.75 ±1.81)%and (59.87 ±4.52)%, respectively.Down-regulation of the KRAS mRNA in Panc-1 cells transfected with siKRAS by scFvCD44v6-IONP-PEI and IONP-PEI was up to (34.02 ±6.15)%and (51.09 ±6.70)%, re-spectively .The protein level of KRAS was lower in targeting group than that in nontargeting group .CONCLUSION:scFvCD44v6-IONP-PEI is a safe and efficient nanoparticulate carrier for gene delivery .It is more effective to transfer siRNA into the cells and mediate gene silencing effect in vitro than the nontargeting group .
8.Effect of placental growth factor gene silencing on migration and invasion of human pancreatic carcinoma cell line PANC1
Jianhua LIU ; Dong MA ; Shaojie CHEN ; Guoda LIAN ; Jiajia LI ; Kaihong HUANG
Chinese Journal of Pancreatology 2016;16(3):159-163
Objective To explore the effect of inhibiting placental growth factor ( PIGF ) by small interfering RNA ( siRNA) on migration, invasion and chemoresistance of human pancreatic cancer cell line PANC1.Methods Three specific siRNAs targeting PIGF (siRNA-PIGF) were designed.PANC1 cells were transfected with siRNA-PIGF by liposome transfection using untransfected cells as blank controls and nonspecific siRNA ( siRNA-NC) transfected cells as negative controls .The PIGF mRNA and protein expression was examined by real-time RT-PCR and ELISA.MTT method was used to assess the inhibition rate of chemotherapeutic reagents on cell proliferation .The abilities of migration and invasion were evaluated by Transwell assay.Results The inhibition rate of PIGF mRNA in PANC1 cells transfected by 3 siRNA-PIGF were (64.38 ±8.92)%, (70.48 ±7.72)% and (81.25 ±6.02)%, which was lowest in siRNA-PIGF-3 transfected cells.The expression of PIGF mRNA in PANC1 cells were decreased by (63.72 ±8.20)%at 24 h after siRNA-PIGF transfection compared with siRNA-NC transfected cells;and the level of PIGF protein in the supernatant of cultured PANC1 cells was lowered by (42.92 ±1.34)% compared with siRNA-NC transfected cells and by (46.25 ±3.64)% compared with untransfected cells at 48h after transfection, which all had significant difference .There was no statistical difference between untransfected and siRNA-NC transfected cells.After 3 ng/L gemicitabine treatment , the inhibition rate of cell proliferation in siRNA-PIGF group was even higher than that in siRNA-NC and untransfected group [(44.35 ±5.05)% vs(34.29 ±3.60)% and (31.01 ±1.08)%;both P<0.05], and no significant difference was not observed after 5-FU and adriamycin treatment.In migration and invasion assay , the number of transmembrane cells from siRNA-PIGF group was 38.1%and 28.2%of that from siRNA-NC group and 40.8% and 36.2% of that from untransfected group , which had statistical difference (all P<0.05).Conclusions PIGF silencing could significantly suppress the migration and invasion of PANC 1 cells and improve the sensitivity to gemicitabine .
9.Association of MYH9,STAT4 and uPA gene single nucleotide polymorphism with idiopathic membranous nephropathy
Yu GUO ; Lu LI ; Kaihong GU ; Zongfeng ZHAO ; Dan XU ; Chen LU
The Journal of Practical Medicine 2016;32(3):376-381
Objective To investigate thecorrelation between nonmusle myosin heavy chain 9 gene (MYH9) rs12107,signal transducer and activator of transcription (STAT4) rs3024912, Urokinase plasminogen activator (uPA) rs4065 single nucleotide polymorphism and idiopathic Uighur membranous nephropathy (IMN). Methods Patients admittedby People′s Hospital of Xinjiang Uyghur Autonomous Region from June 2011 to May 2015 were selected in the research,of which 45 with IMN (group A),45 patients with IgA nephropathy (group B) and 45 healthy controls(group C). The polymorphisms of rs12107,rs3024912 and rs4065 were measured with direct sequencing, in order to analyzing the correlation between genotype and allele with IMN. Results Group Ars12107 (MYH9) locus genotype CC, C allele (48.9%, 65.6%) frequency were higher than those in group B (13.3%, 33.3%) and group C (20.0%, 46.7%), and the difference was statistically significance (P < 0.05). C allele carriers of the risk of IMN is 2.18 times that of the T allele (95% CI: 1.19-3.97). Univariate Logistic regression analysis of rs12107 CC genotype showed patients with CC genotype faced with high risk of renal failure (OR = 5.56,95% CI:1.27-24.29, P = 0.023) compared with non-CC genotype patients. rs3024912 genotype and allele frequencies showed no significant difference among the three groups (P > 0.05). rs3024912 GG genotype patients showed higher risk of renal failure compared with non-GG genotype patients (95% CI:1.48-26.83, P = 0.013). Only TT genotype was detected on rs4065 locus. TC and CC genotype were not detected. Conclusions MYH9 gene rs12107 locus CC genotype and C allele are associated with susceptibility to IMN in Xinjiang Uygur, and CC genotypes associated with renal function. rs3024912 (STAT4)GG genotype are not susceptibility gene,but associated with renal function in patients with IMN.
10.Hematoporphyrin derivative photodynamic therapy of human pancreatic cancer cells in vitro
Zhong YU ; Kaihong HUANG ; Lingyun WANG ; Zhaohua ZHU ; Rufu CHEN ; Jinxin ZHANG
Chinese Journal of Pancreatology 2010;10(1):34-36
Objective To investigate the killing effect of hematoporphyrin derivative photedynamic therapy (PDT) on cultured human pancreatic cancer cell,and to explore the mechanism of this effect.Methods Biolitec PDT 630 semi-conductor laser therapeutic apparatus was used as the light source.After pancreatic cancer cell PANC1 was incubated 8 h with different concentrations of Photosan(hematoporphyrin derivative) as photosensitizer (0.5mg/L,1 mg/L,2 mg/L,4 mg/L),the cells were given different doses of 630nm laser irradiation(1 J/cm2' 5 J/cm~2,10 J/cm~2 ).The A492 value was determined in each group with MTT method.Cell apoptosis rate was detected by flow cytometry after PDT.Results There was no killing effect when no Photosan was administrated;10 J/cm~2 irradiation had killing effect on PANC1 when Photosan was administrated as 1 mg/L(0.140±0.013 vs 0.213±0.008,P<0.05);5 and 10 J/cm~2 irradiation all had killing effect on PANC1 when Photosan was administrated as 2 mg/L (0.081±0.024 and 0.049±0.013vs 0.211±0.031,P<0.05 and P<0.01 );all doses of irradiation had killing effect when Photosan was administrated as 4 mg/L.There was no significant difference between 5 and 10 J/cm~2 irradiation in term of killing effect.Cell apoptosis rates with 0 or 2 or 4 mg/L Photosan and 10 J/cm~2 irradiation were(13.8±1.8) %,(40.9±1.6)%,(62.5±2.0)%,the difference was statistically significant(P<0.05).Conclusions Photosensitizer or irradiation alone did not produce PDT effect.With certain dose of photosensitizer and irradiation,the PDT effect increased accordingly.