1.Pharmacokinetic study of perlolyrine in rats by means of stable isotope dilution
Ganghua TANG ; Guohui JIANG ; Lianfang ZHENG
Chinese Pharmaceutical Journal 2001;(1):45-47
OBJECTIVE To determine the pharmacokinetic parameters of perlolyrine in rats.METHODS The plasma concentration and pharmacokinetic parameters of perlolyrine were determined by means of GC-MS with selected ion (m/z 247 and m/z 248) and [2-15N]perlolyrine(m/z 248) as the internal standard.RESULTS The concentration-time profile of perlolyrine after oral administration of perlolyrine fitted a two-compartment open model.The pharmacokinetic parameters were t1/2α=0.31 h, t1/2β=4.62 h, t1/2ka=0.10 h, tmax=0.34 h, cmax=18.74 ng.mL-1,K12=0.82 h-1, K21=0.38h-1,K10=0.29 h-1,Vb=108.16 L.kg-1,AUC=98.54 ng.h.mL-1,respectively.CONCLUSION The method is constant,sensitive and accurate.It provided a scientific basis for the clinical use of perlolyrine.
2.rAAV2-EGFP transfer to rat retina mediated by ultrasound and SonoVue after intravitreal and subretinal injection
Hongli LI ; Lianfang DU ; Xiaozhi ZHENG ; Huiping WANG ; Jin QIAN ; Feng WANG ; Huiming LI
Chinese Journal of Ultrasonography 2010;19(5):440-442
Objective To investigate the practical efficacy and safety of ultrasound with microbubbles mediated rAAV2-EGFP to retina of rat after intravitreal and subretinal injection. Methods Gene transfer was examined by rAAV2-EGFP intravitreal and subretinal injection into the Wistar rats with or without microbubbles. The eyes were exposed to US (1 MHz,2 W/cm2, duration 5 minutes,duty cycle 50%,pulse recurrent frequency 100 Hz). The onset of EGFP gene expression, lightness of fluorescence, area of fluorescence and its distribution in the fundus in vivo via fluorescence stereosocope were investigated on the 4th,7th, 35th,49th and 120th day respectively. The value of gene transfer was quantified through the EGFP fluorescence quantitative methods by Axiovision 3. 1 software. HE staining was used to observe tissue damage. Results There was no fluorescence observed by fluorescence stereosocope after intravitreal injection after two-month study. After subretinal injection, ultrasound-targeted microbubbles destruction (UTMD) strongly increased gene transfer efficiency. UTMD used in the experiment did no harm to the rat retina structure. Conclusions UTMD could not enhance rAAV2-EGFP transfecion efficiency to rat retina after intravitreal injection but the transduction could be enhanced significantly after subretinal injection.
3.Inhibition effect of anti-epidermal growth factor receptor antibody mediated by recombinant adeno-associated virus vector on pancreatic cancer cell lines
Shiqi WAN ; Taiping ZHANG ; Tianxiao WANG ; Lianfang ZHENG ; Fang LI ; Huihua LI
Chinese Journal of Digestive Surgery 2011;10(4):286-289
Objective To construct a recombinant adeno-associated virus(rAAV)vector containing a human anti-epidermal growth factor receptor(anti-EGFR)single-chain variable fragment antibody gene,and observe its inhibitory effects on pancreatic cancer cell lines.Methods Human anti-EGFR single-chain variable fragment antibody gene was inserted into the Kpn I and Bgl Ⅱ sites to construct a rAAV-anti EGFR vector,and then rAAV1-EGFP group and rAAV1-anti EGFR group were established.The expression of anti-EGFR antibody was observed.Antibody expression was detected by Western blot,and the inhibition and apoptosis rates of human pancreatic cancer cell lines(PCT-3,SW1990,Capan-1,ASPC-1,MiaPaCa-2 and PANC-1 cells)were detected by CCK-8 assay and flow cytometry,respectively.All data were analyzed using the t test.Results The results of Western blot assay demonstrated that anti-EGFR antibody was expressed in 6 pancreatic cancer cell lines.The inhibition rates of rAAV1-EGFP and rAAVl-anti EGFR on pancreatic ASPC-1 cells were 1.1%± 2.4% and 15.1%±3.5%,respectively,with a significant difference between the 2 groups(t =6.598,P <0.05).The apoptosis rates of PANC-1 cells were 7.0% ± 3.0% in the rAAV1-EGFP group and 1 1.4% ± 2.5% in the rAAV1-anti EGFR group,with no significant difference between the 2 grouvs(t = 1.952,P >0.05).The apoptosis rates of SW1990,ASPC-1,Capan-1,PCT-3,MiaPaCa-2 cells were 1.1% ± 0.8%,1.5% ± 0.7%,1.7% ± 1.2%,1.1%±0.7% and 2.2% ± 1.1% in the rAAV1-EGFP group,and 17.6% ± 2.2%,46.9% ± 3.9%,20.0% ±2.8%,12.1% ± 1.6% and 31.1% ±2.5% in the rAAV1-anti EGFR group,respectively,with significant differences between the 2 groups(t = 12.208,19.846,10.405,10.909,18.327,P <0.05).Conclusions A rAAV-anti EGFR vector with human anti-EGFR single-chain variable fragment antibody gene was constructed.Anti-EGFR antibody has obvious inhibition effects on pancreatic cancer cell lines.
4.The combination of rAAV-anti EGFR with gemcitabine and radiation in pancreatic cancer
Tianxiao WANG ; Jianwei XU ; Lianfang ZHENG ; Shiqi WAN ; Taiping ZHANG ; Yupei ZHAO
Chinese Journal of Hepatobiliary Surgery 2013;(4):291-295
Objective EGFR targeted therapy mediated by adeno-associated virus is a promising way to treat pancreatic cancer.This study aimed to assess the feasibility and activity of combining rAAV-anti EGFR,gemcitabine,and radiation in pancreatic cancer cells.Methods Aspc-1 human pancreatic carcinoma cells were divided into several groups,in vitro and in vivo,which were respectively exposed to gemcitabine alone,radiation alone,rAAV-anti EGFR alone,the combination of rAAV-anti EGFR with gemcitabine,the combination of rAAV-anti EGFR with radiation,and the combination of all three agents.The pancreatic cancer tumor growth and apoptotic rate were measured.Results The apoptotic rate was higher in cells treated with a single or combination of agents compared to the negative control (P<0.05).The combination of rAAV-EGFR,gemcitabine,and radiation produced the highest induction of apoptosis compared to a single agent alone (P < 0.05).Treatment with rAAV-anti EGFR greatly inhibited growth in the tumor xenografts (P<0.05),and a synergistic effect of rAAV-anti EGFR,gemcitabine,and radiation was found.The number of tissue cancer cells that expressed cleaved caspase-3 after treatment with rAAV EGFR was more than that of the control group (P<0.05).The combined treatment of rAAV-anti EGFR,gemcitabine,and radiation induced the highest numbers of cells expressing cleaved caspase-3 compared to that with a single agent alone (P<0.05).Conclusions The rAAV-anti EGFR therapy in combination with chemotherapy and radiation therapy demonstrated a greater efficacy over therapy with a single agent alone.rAAV-anti EGFR increased the efficacy of gemcitabine and radiation in the treatment of pancreatic cancer cells.
5.Ultrasound-targeted microbubble mediated liposome small interference RNA transfection to retinal pigment epithelium cells
Hongli LI ; Lianfang DU ; Xiaozhi ZHENG ; Huiping WANG ; Qing GU ; Xiafang CHEN
Chinese Journal of Medical Imaging Technology 2010;26(1):22-24
Objective To investigate the transfection efficiency of combination of ultrasound (US) microbubbles (MBs) mediated liposome small interference RNA (siRNA) to human and rat retinal pigment epithelium (RPE) cells. Methods Human and rat RPE cells and siRNA were incubated in 24-well plates (2×10~5/well and 1×10~5/well, respectively). The cells were devided into 5 groups: siRNA+US, siRNA+MBs+US, siRNA+L (liposome), siRNA+L+US, siRNA+L+US+MBs. After 12 h, transfection efficiency was observed with fluorescence microscopy and flow cytometry. Results US or ultrasound targeted microbubbles destruction without liposome-mediated could not promote siRNA transfection efficiency to human and rat RPE cells. The transfection efficiency of human and rat RPE cells significantly decreased in the siRNA+L+US+MBs group, but increased in siRNA+L+US group. Conclusion Ultrasonic irradiation can promote lipid-mediated siRNA transfected human RPE cells.
6.The endpoint detection of cough signal in continuous speech.
Guoqing YANG ; Hongqiang MO ; Wen LI ; Lianfang LIAN ; Zeguang ZHENG
Journal of Biomedical Engineering 2010;27(3):544-555
The endpoint detection of cough signal in continuous speech has been researched in order to improve the efficiency and veracity of manual recognition or computer-based automatic recognition. First, using the short time zero crossing ratio(ZCR) for identifying the suspicious coughs and getting the threshold of short time energy based on acoustic characteristics of cough. Then, the short time energy is combined with short time ZCR in order to implement the endpoint detection of cough in continuous speech. To evaluate the effect of the method, first, the virtual number of coughs in each recording was identified by two experienced doctors using the graphical user interface (GUI). Second, the recordings were analyzed by automatic endpoint detection program under Matlab7.0. Finally, the comparison between these two results showed: The error rate of undetected cough is 2.18%, and 98.13% of noise, silence and speech were removed. The way of setting short time energy threshold is robust. The endpoint detection program can remove most speech and noise, thus maintaining a lower rate of error.
Algorithms
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Artificial Intelligence
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Cough
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physiopathology
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Endpoint Determination
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Humans
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Pattern Recognition, Automated
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methods
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Signal Processing, Computer-Assisted
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Sound
7. Correlation between miR-1178 expression and clinicopathological significance in human pancreatic cancer
Zhe CAO ; Suli ZHENG ; Gang YANG ; Mengyu FENG ; Lianfang ZHENG ; Taiping ZHANG ; Yupei ZHAO
Chinese Journal of Surgery 2017;55(6):468-473
Objective:
To test the expression of miR-1178 in pancreatic cancer and study its clinicopathological significance and mechanism.
Methods:
The expression of miR-1178 in 87 paired paraffin pancreatic ductal adenocarcinoma specimens and adjacent non- cancerous pancreatic tissue diagnosed by Pathology Department of Peking Union Medical College Hospital was detected by hybridization in situ. The relationship between the expression of miR-1178 and clinicopathological characters was analyzed.miR-1178 mimics and inhibitor were used to further detect the close relationship among miR-1178 and cancer invasion. Establish a nude mice subcutaneously transplanted tumor model, 4 weeks after vaccination for tumor volume and weight measurement.Student
8. The expression of KLK7 in pancreatic cancer and the effects on the biological behavior of pancreatic cancer cells
Suli ZHENG ; Mengyu FENG ; Gang YANG ; Guangbing XIONG ; Lianfang ZHENG ; Taiping ZHANG ; Yupei ZHAO
Chinese Journal of Surgery 2018;56(5):391-397
Objective:
To investigate the expression of KLK7 in pancreatic cancer and its clinical significance.
Methods:
Immunohistochemistry was used to detect the expression of KLK7 protein in pancreatic cancer tissue microarray with 92 samples. Statistical analysis of the relationship between KLK7 and clinicopathological characteristics was finished. Pancreatic cancer cell lines were infected with lentiviuses in order to get cells with KLK7 stable overexpression.KLK7-siRNA was transfected into pancreatic cancer cells to knock down KLK7.Cell proliferation and chemosensitivity were detected by CCK-8 assay; Cell invasion and migration abilities were detected by Transwell assay. At the same time, subcutaneous xenograft tumor models were established in nude mice to observe the effect of KLK7 on tumor growth in nude mice. Data were statistically analyzed by rank sum test, χ2 test and Logistic regression analysis.
Results:
The expression level of KLK7 in pancreatic cancer tissues was higher than that in paired adjacent tissues (
9.Comparison of prosthesis locations and postoperative hip functions between supercapsular percutaneously-assisted total hip (SuperPATH) and posterolateral approaches in total hip arthroplasty
Weicheng ZHANG ; Feng ZHU ; Kai ZHENG ; Mingzhou WU ; Lianfang ZHANG ; Jun ZHOU ; Rongqun LI ; Yaozeng XU
Chinese Journal of Orthopaedic Trauma 2021;23(7):571-576
Objective:To compare the prosthesis locations and postoperative hip functions between supercapsular percutaneously-assisted total hip (SuperPATH) approach and traditional posterolateral approach (PLA) in total hip arthroplasty.Methods:A retrospective analysis was conducted of the 107 patients who had undergone unilateral total hip arthroplasty at Department of Orthopedic Surgery, The First Affiliated Hospital to Soochow University from August 2016 to February 2019. They were divided into 2 groups according to their surgical approaches. In the SuperPATH group of 54 cases, there were 20 males and 34 females with an age of (64.3±9.1) years; in the PLA group of 53 cases, there were 20 males and 33 females with an age of (62.2±10.6) years. The 2 groups were compared in terms of abduction angle, ratio of abduction angle to safety zone, anteversion angle, ratio of anteversion angle to safety zone, retroversion angle, incidence of retroversion, and differences in eccentricity and lower limb length on the first day after operation, and Harris hip scores at 1 week, 3 months and the last follow-up postoperatively. Their complications were also recorded as well.Results:There were no statistically significant differences in the preoperative general data between the 2 groups, showing comparability ( P>0.05). The postoperative imaging data were complete for the 107 patients who had been followed up satisfactorily for 14 to 36 months (average, 25 months). The SuperPATH group had significantly larger retroversion angle (13.6°±9.6°) and incidence of retroversion (18.5%, 10/54), significantly smaller difference in eccentricity [0.26 (0.13,0.49) cm], and significantly higher Harris hip score [(74.8±7.8) points] at one week after surgery than those in the PLA group [3.0°±1.0°; 5.7%, 3/53; 0.38 (0.13,0.70) cm; (72.0±6.7) points] ( P<0.05). There were no statistically significant differences between the 2 groups in abduction angle, ratio of abduction angle to safety zone, anteversion angle, ratio of anteversion angle to safety zone, difference in lower limb length, or Harris hip scores at 3 months or the last follow-up postoperatively (all P>0.05). Follow-ups in both groups observed no more than one case of dislocation which responded to manual reduction. Conclusion:The minimally invasive SuperPATH approach may obtain better femoral eccentricity and higher early hip function scores than the traditional posterolateral approach, but may lead to a higher incidence of retroversion after prosthesis placement.
10.The long coding RNA GSTM3TV2 acts an oncogene to promote chemoresistance in pancreatic cancer
Guangbing XIONG ; Gang YANG ; Mengyu FENG ; Fangyu ZHAO ; Lianfang ZHENG ; Taiping ZHANG ; Yupei ZHAO
Chinese Journal of Surgery 2019;57(9):691-697
Objectives To examine the expression of the long coding RNA GSTM3TV2 in pancreatic cancer tissues and to examine its role and mechanism in chemoresistance of pancreatic cancer cells. Methods The expression of lncRNA GSTM3TV2 in 15 pancreatic cancer specimens and corresponding adjacent to cancer tissue samples diagnosed by Department of Pathology, Peking Union Medical College Hospital was detected by real?time PCR.And the expressions of GSTM3TV2 in pancreatic cancer cell AsPC?1,BxPC?3,MIAPaCa?2,PanC?1,SU86.86,T3M4,and chemoresistant cells AsPC?1/GR and MIAPaCa?2/GR, and human pancreatic nestin?expressing cells hTERT?HPNE were detected. Pancreatic cancer cell lines were transfected with GSTM3TV2?pcDNA3.1(+)in order to get cells with GSTM3TV2 overexpression.GSTM3TV2?siRNA was transfected into pancreatic cancer cells to knock down GSTM3TV2. The cell chemoresistance was measured by CCK?8 and flow cytometry assay when incubated with nab?paclitaxel. At the same time, subcutaneous xenograft tumor models were established in nude mice to observe the effect of GSTM3TV2 on chemoresistance of tumor growth in nude mice.Western blot assay was also performed to detect the molecular mechanism of chemoresistance of GSTM3TV2. Results Comparing toadjacent tissues(0.084 ± 0.019), GSTM3TV2 expression was significantly upregulated in the pancreatic cancer tissues(0.493 ± 0.084) (t=5.146, P<0.05). GSTM3TV2 expression were higher in the chemotherapy resistance pancreatic cancer cells AsPC?1/GR(210.799±19.788) and MIAPaCa?2/GR(122.408±23.419) than that in the AsPC?1(3.793±0.615) and the MIAPaCa?2(5.179±1.095)(t=21.800,P<0.05;t=-18.490,P<0.05). The results of in vivo experiments showed that the volume of subcutaneously transplanted tumors in the overexpressing GSTM3TV2 group ((1 059.609±102.498)mm3) was significantly larger than that in the control group((566.414±81.087) mm3) by treated with nab?paclitaxel(t=4.230,P<0.05).Meanwhile,GSTM3TV2 could promote the expression of Cyclin D1, CDK6, Cyclin E1, Vimentin, N?cadherin, ZEB1, Snail and Slug; but decrease cleaved caspase?3,cleaved PARP in pancreatic cancer cells.Conclusions The expression level of GSTM3TV2 in pancreatic canceris higher than that in paired adjacent tissues. GSTM3TV2 may act as an oncogene to promote chemoresistance in pancreatic cancer through regulation of cell proliferation,apoptosis, and epithelial?mesenchymal transition.