1.Effects and underlying mechanism of siRNA targeting PARP1 on the proliferation of PC3 cells
Wenqi WU ; Zhenzhen KONG ; Hanliang ZHU ; Xiaolu DUAN ; Shujue LI
Chinese Journal of Urology 2013;(2):130-134
Objective To observe the effects and study the underlying mechanism of siRNA targeting PARP1 on the proliferation of androgen independent prostate cancer PC3 cell line.Methods Three specific siRNA sequences targeting PARP1 were designed and synthesized.And two sequences which had better interfering effect on the expression of PARP1 were evaluated and selected through lipofectamine transfection,RT-PCR and Western Blot.The effect of PARP1 silencing on the proliferation of PC3 cells was observed with MTS assay and the levels of the phosphorylation of Akt and GSK3β were detected by Western Blot.Results Compared to the blank control group,the transfected group with the negative control sequence had no significant impact on the expression of PARP1,however the transfected group with siRNA-1706,-2003 or-2907 could significantly suppress the mRNA and protein expression of PARP1.The mRNA inhibition rate reached to(52.07 ± 4.65)%,(44.38 ± 9.15)% and(22.05 ± 6.65)%,respectively;and the protein inhibition rate reached to(86.86 ± 4.94)%,(83.30 ± 7.18)% and(63.05 ± 10.19)%,respectively.The siRNA-1706 and-2003 could significantly inhibit the proliferation of PC3 cells;the inhibition rate was(38.93 ± 3.87)% and(34.93 ± 1.21)%.And they also could down-regulate the intracellular levels of phosphorylated Akt and GSK3β in PC3 cells.Conclusion PARP1-targeted siRNA can significantly suppress the expression of endogenous PARP1 and inhibit the proliferation of PC3 cells,which is related to the inhibition of Akt activity and the activation of GSK3 β.
2.An investigation of life value and suicide attitude of college students in the earthquake-stricken area
Yan ZHANG ; Fanchang KONG ; Zhenzhen QUAN ; Fang XU
Chinese Journal of Behavioral Medicine and Brain Science 2010;19(7):641-643
Objective To understand the life attitude of college students in earthquake-stricken area. Methods 166 college students were conducted by life value scale (LVS) and suicide attitude questionnaire (QSA). Results ①On the LVS, males( (1.85 ±0.57) , (1.73±0.55) ) got significantly higher scores than females ((1.66±0.43),(1.54±0.40)) on survival values-oriented and spiritual values-oriented, and lower scores on tool values-oriented ((2.59±0.46 vs (2.76±0.51) ) , there was significant difference on overcome values-oriented between different grades(F = 3.710, P = 0.013). On the QSA, males (3.01 ±0.51) got significantly lower scores than females (3.28 ±0.51) on the cognition of suicidal behavior difference (t=3.110, P=0.002). There were significant differences on attitude to relationship of suicides between different gender and areas (P<0.05) , and attitude to euthanasia between different gender, grades and subjects (P<0.05). ②Social valuesoriented can predict cognition of suicidal behavior, personal and tool values-oriented can predict attitude to relationship of suicides, the total of life value can predict cognition of suicidal behavior and attitude to relationship of suicides. Conclusion The life value can predict the suicide attitude of college students in earthquake-stricken area, thus the life education should be enhanced to improve their quality of life.
3.Value of three-dimensional power Doppler ultrasound in diagnosis of cervical cancers
Wei LIU ; Litao SUN ; Nana LIU ; Zhenzhen WANG ; Xinying DOU ; Xianchao KONG
Chinese Journal of Ultrasonography 2013;22(12):1056-1059
Objective To evaluate the clinical value of three-dimensional power Doppler ultrasound (3D PDUS) in diagnosing cervical cancers.Methods 162 patients with cervical lesions were enrolled.All the cases were confirmed by autopsy or surgery.Vascularization index (VI),flow index (FI) and vascularization flow index(VFI) were all measured and compared.The pathological findings was taken as golden standard.ROC curve was used to find the best cut-off value.Results 3D PDUS indices of benign and malignant lesions were statistically different (P <0.0001).The best cut-off value of VI was 4.63 using ROC curves,the area under the curve (AUC) was 0.94,the sensitivity and specificity were 89.10% and 86.30%,respectively.The best cut-off value of FI was 34.39 using ROC curves,AUC was 0.76,the sensitivity and specificity were 73.40% and 76.70%,respectively.The best cut-off value of VFI was 1.60 using ROC curves,AUC was 0.93,the sensitivity and specificity were 87.50 % and 86.30 %,respectively.Conclusions 3D PDUS was a new and noninvasive way for the clinical to differentiate benign and malignant cervical lesions.
4.Effect of autophagy inhibitor chloroquine on the renal calcium oxalate crystals formation in rats
Xin MAI ; Zhenzhen KONG ; Tuo DENG ; Zhou YANG ; Yang LIU ; Yu LAN ; Xiaolu DUAN ; Wenqi WU ; Guohua ZENG
Chinese Journal of Urology 2017;38(7):542-547
Objective To investigate the effect and potential mechanism of autophagy inhibitor chloroquine on the calcium oxalate crystals formation in rats.Methods From September 2016 to October 2016,Thirty healthy male SD rats were randomly divided into 3 groups:control group,model group and chloroquine intervention group.The method to establish calcium oxalate stone model was drinking water with 1% ethylene and 1% ammonium chloride freely.The rats of chloroquine intervention group were treat with chloroquine (40mg/kg · d) by intraperitoneal injection.Modeling was finished after 28 days.The amounts of renalcalcium oxalate crystals were detected by polarizing microscope.For all groups,the amounts of autophagosome were detected by transmission electron microscope.Twenty four hour urine compositions for stone risk factors were detected.The expressions of oxidative stress injury related molecular markers (SOD,MCP-1 and 8-OHdG) and the expressions of autophagy markers (LC3 and P62) were detected by immunohistochemistry.The RNA expressions of SLC26A6 in kidney were detected by Real-time PCR.Results Compared to the model group,the amounts of renal calcium oxalate crystals were significantly reduced in chloroquine intervention group (32.37 ± 5.14 vs.4.18 ± 0.25,P < 0.05).Compared to the control group,the level of autophagy was increased in the model group.Compared to the model group,the level of autophagy was inhibited in the chloroquine intervention group.For control group,model group and chloroquine intervention group,the excretion of urinary oxalate were (3.1 ± 1.5) mmol,(22.5 ± 8.1) mmol,(2.8 ± 1.2) mmol,respectively;the excretion of urinary citrate were (63.4 ± 7.4) mmol,(45.9 ± 9.5)mmol,(15.6 ± 8.2) mmol,respectively.Compared to the control group,the amounts of urinary oxalate weresignificantly elevated in model group (P < 0.05),but citrate were significantly reduced in the chloroquineintervention group(P < 0.05).For control group,model group and chloroquine intervention group,theexpressions of SOD were 42.24 ±4.16,19.21 ± 2.25,39.08 3.53,respectively;the expressions of MCP-1 were 4.02 0.51,8.45 ± 0.55,5.52 ± 0.34,respectively;the expressions of 8-OHdG were 7.16 ± 0.54,11.21 ± 1.12,8.67 ±0.34,respectively;the RNA expressions of SLC26A6 were 0.35 ±0.07,1.02 ±0.17,0.70 ± 0.06,respectively.Compared to the control group,the expressions of SOD were significantly reduced in the model group,but the expressions of MCP-1,8-OHdG and SLC26A6 were significantly elevated(P <0.05).Compared to the model group,the expressions of SOD were significantly elevated chloroquine intervention group (P < 0.05),but the expressions of MCP-1,8-OHdG and SLC26A6 were significantly elevated(P < 0.05).Conclusions The autophagy inhibitor chloroquine could inhibit the formation of calcium oxalate crystals induced by ethylene in rat kidney via inhibit the renal autophagy level and expressions of the SLC26A6,reducing the renal oxidative stress injury and urinary oxalate excretion.
5.Detection of chromosome aneuploidies in spontaneous abortion villus samples by quantitative fluorescence PCR.
Zhenzhen WU ; Ning LIU ; Yongjiang ZHAO ; Zhenhua ZHAO ; Xiangdong KONG
Chinese Journal of Medical Genetics 2016;33(2):227-230
OBJECTIVETo assess the value of quantitative fluorescence polymerase chain reaction (QF-PCR) for the detection of chromosomal aneuploidies in chorionic villus samples from early abortion.
METHODSOne hundred seventy seven specimens were collected. Genomic DNA was extracted, and aneuploidies of 8 chromosomes (13, 15, 16, 18, 21, 22, X and Y) were detected by QF-PCR analysis.
RESULTSThe QF-PCR was successful in 176 (99.4%) of the cases. All detection was completed in 48 hours. Sixty three(35.8%) cases have shown abnormal signals, which included 3 cases of trisomy 13, 3 cases of trisomy 15, 14 cases of trisomy 16, 2 cases of trisomy 18, 7 cases of trisomy 22, 3 cases of trisomy 21, 13 cases of 45,X, 1 case of 47,XXX, 2 cases of 47,XXY, 2 cases of haploidy, 11 cases of triploidy, 1 case of trisomy 16 and trisomy 22, 1 case of trisomy 21 and trisomy 22. Trisomy 16 was the most common chromosome aneuploidy (22.22%), which was followed by 45,X (20.63%), triploidy (17.46%) and trisomy 22 (11.11%).
CONCLUSIONQF-PCR is a quick and easy method for detecting chromosomal aneuploidies in chorionic villi tissue. The results can provide important information for genetic counseling for spontaneous abortions.
Abortion, Spontaneous ; diagnosis ; genetics ; Adult ; Aneuploidy ; Chorionic Villi ; chemistry ; Female ; Fluorescence ; Humans ; Pregnancy ; Prenatal Diagnosis ; Real-Time Polymerase Chain Reaction ; methods
6.Immunological regulation of CD70/CD27 pathway and the expression of CD70 in tumor
Zhenzhen LIU ; Yingjun KONG ; Wenxue SUN ; Shimin LI ; Min ZHANG ; Hesong YIN
Journal of International Oncology 2018;45(8):487-489
CD70/CD27 pathway plays an important role in human immune regulation.The role of CD70/CD27 pathway in immune regulation is mainly to promote the activation and proliferation of T cells,induce the differentiation and formation of effector T cells and memory T cells,and interfere with regulatory T cells.In addition,the high level of CD70 expression in some tumor cells provides a new way for tumor immunotherapy.