1.Effect of knockdown of ARHGAP30 on proliferation and apoptosis of Siha cells
Ya-Ting PENG ; Duan LIU ; Jie MENG ; Wen-Chao LI ; Hui-Qi LI ; Hua GUO ; Mei-Lan NIU ; Qiao-Hong QIN
Chinese Pharmacological Bulletin 2024;40(5):847-853
Aim To investigate the changes in the proliferation and apoptosis of Siha cells after knocking down Rho GTPase-activating protein 30(ARHGAP30).Methods After designing specific shARHGAP30 primers and connecting them to the pLKO.1 vector,we transformed them into Escherichia coli competent cells,then co-transfecting them with lentiviral helper plasmids into HEK-293T cells.We collected and filtered cell supernatant to obtain the vi-rus to infect Siha cells.RT-qPCR and Western blot were used to detect knockdown efficiency,as well as changes in the expression of Bax and Bcl-2 after trans-fection.The CCK-8 method was employed to measure the proliferation level of cells after knockdown.Results After successful construction of a lentiviral plasmid with knockdown of the ARHGAP30 gene and establish-ment of stably transfected Siha cells,ARHGAP30 tran-scription and translation(P<0.01)in Siha cells de-creased,Bax/Bcl-2 significantly decreased(P<0.01),indicating decreased apoptosis and increased cell proliferation(P<0.01).Conclusions This study suggests the involvement of ARHGAP30 in the proliferation and apoptosis of Siha cells,and regulating the ARHGAP30 gene may interfere with the occurrence and development of cervical cancer.
2.A study on reversing multidrug resistance of MCF-7/ADR cells by inhibiting doxorubicin efflux with a fluorinated nanomedicine delivery system
Huai-Tao SUN ; Yin-Hua LIN ; Yu-Rong SUN ; Qiao CHEN ; Shuo CHEN ; Yu CAO
Chinese Pharmacological Bulletin 2024;40(6):1124-1130
Aim To achieve efficient loading of doxo-rubicin(doxorubicin,dox)by synthesizing fluorinated nanomaterials,and to study the mechanism of nano-medicine against multidrug resistance in tumors.Methods The physicochemical properties of nano dox(nano-doxorubicin)at different degrees of fluorination(PF4 and PF8)were evaluated through various physi-cochemical parameters,and nano dox formulations were screened out with superior fluorination.The mechanism of nano dox in overcoming multidrug resist-ance in tumors was investigated and in vitro anti-tumor activity was studied through research on cytotoxicity,cellular uptake,apoptosis,and resistance against the P-glycoprotein(P-glycoprotein,P-gp)efflux pump.Results By examining various physicochemical indi-cators,PF8 exhibited superior nano performance in loading doxorubicin compared to PF4.Cytotoxicity ex-periments demonstrated that the IC50 value of nano dox was only 1/40 of free dox,indicating excellent in vitro activity of nano dox in overcoming resistance and inhib-iting cell proliferation.Cell uptake and apoptosis stud-ies indicated that nano dox could effectively enhance the uptake of doxorubicin in MCF-7/ADR cells and successfully enter the cell nucleus,ultimately leading to apoptosis-induced anticancer activity.Studies on drug efflux mediated by P-glycoprotein indicated that nano dox could rapidly enter MCF-7/ADR cells through an efficient cellular uptake pathway,effective-ly bypassing the P-glycoprotein efflux pump.This dual action resulted in increased drug uptake within resistant cells and a reduction in efflux,demonstrating the po-tential of nano doxorubicin to overcome drug resist-ance.Conclusions The design of fluorinated nano dox effectively circumvents the efflux of drug molecules by P-glycoprotein,enhancing drug uptake within re-sistant tumor cells and reducing efflux.This results in an effective strategy in combating multidrug resistance in tumors.
3.Effect mechanism of dihydromyricetin on the tumor biological characteristics of endometrial cancer cells
Wei-Hua CUI ; Jing-Ying SONG ; Shu-Xin QIAO ; Shu-Zhong DUAN
Journal of Regional Anatomy and Operative Surgery 2024;33(12):1039-1043
Objective To explore the effects of dihydromyricetin on the proliferation,apoptosis,and invasion of endometrial cancer(EC)cells and its possible mechanisms.Methods Ishikawa cells in the logarithmic growth phase were taken and divided into the control group,20 μmol/L dihydromyricetin intervention group,40 μmol/L dihydromyricetin intervention group,and 80 μmol/L dihydromyricetin intervention group,which were treated with different final concentrations of dihydromyricetin(0 μmol/L,20 μmol/L,40 μmol/L,80 μmol/L).Then,CCK-8 assay and flow cytometry were used to detect the effects of dihydromyricetin on the cell proliferation and apoptosis.Transwell experiment was used to detect the effect of dihydromyricetin on the cell invasion.qRT-PCR and Western blot were used to detect the effects of dihydromyricetin on the cell expression of miR-21 and PTEN.Results Compared with the control group,the cell proliferation inhibition rate and apoptosis rate in the dihydromyricetin intervention group were significantly increased(P<0.05),and gradually increased with the increase of dihydromyricetin concentration(P<0.05).Compared with the control group,the numbers of migration and invasion cells in the dihydromyricetin intervention group were significantly decreased(P<0.05),and gradually decreased with the increase of dihydromyricetin concentration(P<0.05).Compared with the control group,the cell expression of miR-21 in the dihydromyricetin intervention group was significantly decreased(P<0.05),and gradually decreased with the increase of dihydromyricetin concentration(P<0.05),the expression levels of PTEN mRNA and protein were significantly increased(P<0.05),and gradually increased with the increase of dihydromyricetin concentration(P<0.05).Conclusion Dihydromyricetin can inhibit the growth and metastasis of EC cells,and the inhibitory effect is positively correlated with its concentration.The mechanism may be related to the effect of dihydromyricetin on the miR-21/PTEN signaling pathway of EC cells.
4.Correlation Analysis between Serum Fibronectin 3 Levels and Early Severe Bleeding in Patients with Newly Diagnosed Acute Promvelocvtic Leukemia
Hong-Bin ZHAO ; Jia-Jia QIAO ; Xue-Hua HE
Journal of Experimental Hematology 2024;32(4):1058-1062
Objective:To analyze the correlation between serum fibronectin 3(Ficolin-3)levels and early severe bleeding in newly diagnosed acute promyelocytic leukemia(APL)patients.Methods:A total of 125 patients with newly diagnosed APL admitted to Shanxi Bethune Hospital from January 2020 to August 2023 were selected.All patients were given all-trans retinoic acid+arsenic for induction therapy.The severe bleeding events within 30 days of induction therapy(assessed by WHO bleeding score,grade 0,grade 1 and grade 2 were no bleeding or mild bleeding,grade 3 and grade 4 were severe or fatal bleeding)were used as observation endpoints.The serum Ficolin-3 levels was dected by ELISA method,baseline data and other laboratory indicators were counted,and the correlation between serum Ficolin-3 levels and early severe bleeding in newly diagnosed APL patients was analyzed.Results:23 out of 125 APL patients experienced early severe bleeding during induction therapy,including 13 cases of grade 3 bleeding and 10 cases of grade 4 bleeding.There were 102 cases of non-serious bleeding,including 30 cases of grade 0,24 cases of grade 1 bleeding,and 48 cases of grade 2 bleeding.The proportion of serum promyelocytes,white blood cell count,and D-D level in the severe bleeding group were significantly higher than those in the non severe bleeding group(P<0.05),while the levels of PLT and FIB were significantly lower than those in the non-serious bleeding group(P<0.05).The serum Ficolin-3 levels in the severe bleeding group were significantly lower than those in the non severe bleeding group before treatment,7 days of treatment,14 days of treatment,and 30 days of treatment(P<0.05).Confirmed by point two column correlation,serum Ficolin-3 levels were negatively correlated with early severe bleeding in newly diagnosed APL patients before treatment,7 days,14 days,and 30 days after treatment(r values were-0.485,-0.397,-0.304,and-0.183,respectively).The receiver operating characteristic curve(ROC)graph of the subjects was drawn,and the results showed that the area under the curve(AUC)of serum Ficolin-3 levels before treatment and at 7 and 14 days after treatment for predicting early severe bleeding in newly diagnosed APL patients was greater than 0.7,all of which had certain predictive efficacy,and the serum Ficolin-3 level before treatment had the best predictive efficacy.Conclusion:The serum Ficolin-3 levels in newly diagnosed APL patients are associated with early severe bleeding,and the serum Ficolin-3 levels before treatment have a significant advantage in predicting early severe bleeding in newly diagnosed APL patients.
5.The crosstalk of Wnt/β-catenin signaling and p53 in acute kidney injury and chronic kidney disease
Wen-Hua MING ; Lin WEN ; Wen-Juan HU ; Rong-Fang QIAO ; Yang ZHOU ; Bo-Wei SU ; Ya-Nan BAO ; Ping GAO ; Zhi-Lin LUAN
Kidney Research and Clinical Practice 2024;43(6):724-738
Wnt/β-catenin is a signaling pathway associated with embryonic development, organ formation, cancer, and fibrosis. Its activation can repair kidney damage during acute kidney injury (AKI) and accelerate the occurrence of renal fibrosis after chronic kidney disease (CKD). Interestingly, p53 has also been found as a key modulator in AKI and CKD in recent years. Meantime, some studies have found crosstalk between Wnt/β-catenin signaling pathways and p53, but more evidence is required on whether they have synergistic effects in renal disease progression. This article reviews the role and therapeutic targets of Wnt/β-catenin and p53 in AKI and CKD and proposes for the first time that Wnt/β-catenin and p53 have a synergistic effect in the treatment of renal injury.
6.Genetic analysis and PGT-SR outcome of a male carrier of exceptional complex chromosome rearrangement
Dun LIU ; Yun-Qiao DONG ; Chuang-Qi CHEN ; Xing-Su YU ; Jin YAN ; Feng-Hua LIU ; Xi-Qian ZHANG
National Journal of Andrology 2024;30(7):627-633
Objective:To investigate the clinical and genetic characteristics of a male carrier of exceptional complex chromo-some rearrangement(CCR)and the outcome of preimplantation genetic testing for chromosomal structural rearrangement(PGT-SR).Methods:Using the modified high resolution G banding technique and whole-genome low-coverage sequencing(WGLCS),we analyzed the cellular karyotype and molecular karyotype of a male carrier of CCR,performed an analysis of the single-sperm chromosome copy number and conducted PGT-SR for the patient by next-generation sequencing(NGS).In addition,we reviewed the literature on repor-ted male carriers of CCRs and summarized their normal/balanced sperm ratios and PGT-SR outcomes.Results:The karyotype of the patient was 46,XY,der(5)inv(5)(q14.3q23.2)t(5;14;11)(q23.2;q31.1;q21),der(11)t(5;14;11);der(14)t(5;14;11),with the translocation breakpoints located in the intergenic region.Single-sperm sequencing revealed 20.0%(7/35)of normal haploids in the male's spermatozoa,and the results PGT-SR showed a proportion of 25.0%(4/16)of normal/balanced embryos.After thawing and transferring of 2 euploid blastocysts,a healthy male infant was successfully delivered.Conclusion:The proportion of normal hap-loids in the spermatozoa of male CCR carriers may be higher than theoretically predicted,and PGT-SR can effectively improve the preg-nancy outcome in male CCR carriers and provide valuable data for genetic counseling.
7.Chemical constituents from Solanum lyratum and their anti-tumor activities
Lin-Guo ZHANG ; Fu-An WANG ; Man-Hua QIAO ; Li-Ya HAO ; Li LIU
Chinese Traditional Patent Medicine 2024;46(7):2252-2258
AIM To study the chemical constituents from Solanum lyratum Thunb.and their anti-tumor activities.METHODS The extract from S.lyratum was isolated and purified by silica gel column chromatography and Sephadex LH-20,then the structures of obtained compounds were identified by physicochemical properties and spectral data.The anti-tumor activities were determined by MTT mothod.RESULTS Twenty-one compounds were isolated and identified as royleanone(1),dinbutyl phthalate(2),dipentyl phthalate(3),sativan(4),2′,4′-dihydroxy-chalcone(5),isopetasin(6),balanophonin(7),10-eicosenoic acid(8),4,6,7-trimethoxy-5-methyl-coumarin(9),hydroxydihydrobovolide(10),axillarin(11),xanthocyanin(12),magnoflorine(13),verlinic acid(14),apigenin(15),luteolin(16),7-ketositosterol(17),pinoresinol(18),bavachinine(19),bisbenzopyran(20),isotachioside(21).The IC50 values of compounds 6-7,11 and 18-19 ranged from(28.37±3.71)to(73.26±8.61)μmol/L.CONCLUSION All the compounds are first isolated from this plant.Compounds 6-7,11 and 18-19 have certain anti-tumor activities.
8.Analysis of Learning Curve of Bikini Incision Direct Anterior Approach Total Hip Arthroplasty
Zuzhou WEN ; Hongqiang CHEN ; Xizhi WU ; Ang LUO ; Hua ZHOU ; Junzhao QIAO ; Yun YE
Chinese Journal of Minimally Invasive Surgery 2024;24(9):604-609
Objective To explore the learning curve of Bikini incision direct anterior approach total hip arthroplasty(Bikini-THA).Methods Clinical data of 80 cases of Bikini-THA initially completed by an operator skilled in posterolateral approach and direct anterior approach total hip arthroplasty from March 2020 to March 2023 were retrospectively analyzed,and the learning curve was observed by scatter plots of operative time and intraoperative bleeding.They were divided into groups A to D according to the order of surgery,with 20 cases in each group.The operative time,intraoperative bleeding,acetabular abduction angle,anteversion angle,angle between stem and femoral axis,postoperative hip Harris score and complications were compared among the 4 groups.Results After about 40 cases,the decreasing trend of operative time and intraoperative bleeding slowed down and stabilized.The operative time and intraoperative bleeding in the 4 groups were group A>group B>groups C and D(P<0.05),and the differences between the group C and group D were not statistically significant(P>0.05).The acetabular prosthesis was well-positioned in the 4 groups(abduction angle of 30°-50°,anteversion angle of 5°-25°).The femoral prosthesis center fixation rate(angle between stem and femoral axis ≤3°)was group A<groups C and D(P=0.003,0.003).The differences in the Harris scores of the hip joints at 2 weeks,1 month,3 months,and 12 months postoperatively of the 4 groups were not statistically significant(P>0.05),and the efficacy evaluations of the hip joints at 12 months postoperatively were all excellent.There were 5 cases of complications in the group A(2 cases of greater trochanter fracture and 3 cases of lateral femoral cutaneous nerve injury),3 cases of complications in the group B(1 case of greater trochanteric fracture,1 case of lateral femoral cutaneous nerve injury,and 1 case of incision infection),1 case of complications in the group C(lateral femoral cutaneous nerve injury),and no complications in the group D.The follow-up period lasted for 12-26 months,with a mean of(19.4±4.7)months.There were no complications such as dislocation or loosening of the prosthesis.Conclusion The Bikini-THA learning curve was approximately 40 cases.
9.Electroacupuncture combined with Western medication improves lower-limb motor function and blood circulation in patients with cerebral infarction-induced hemiplegia in acute stage:a clinical study
Xuerong YANG ; Lin QIAO ; Jun YAN ; Linxin ZHANG ; Yaya GAO ; Hua GUO ; Jue HONG
Journal of Acupuncture and Tuina Science 2024;22(3):232-238
Objective:To observe the effects of electroacupuncture(EA)combined with Western medication on lower-limb motor function and blood circulation in patients with cerebral infarction-induced hemiplegia in the acute stage. Methods:One hundred eligible patients with acute ischemic stroke accompanied by lower-limb motor dysfunction were allocated to an observation group and a control group using the random number table method,with 50 cases in each group.The control group received routine Western medications for treatment,and the observation group received additional EA intervention.After 2-week and 4-week treatments,the improvement of lower-limb motor function was assessed using the Fugl-Meyer assessment scale for lower extremity(FMA-LE),and changes in the peak blood flow velocities of the posterior tibial(PT)and dorsalis pedis(DP)arteries on the affected side were detected using Doppler. Results:Three cases dropped out during the study,so there were 48 cases in the observation group and 49 in the control group collected for statistical analysis.The FMA-LE score and the peak blood flow velocities of PT and DP arteries increased after 2-week and 4-week treatments in both groups compared with the pre-treatment baseline(P<0.05).After 2-week treatments,the FMA-LE score and the peak blood flow velocities of PT and DP arteries showed no significant differences between the two groups(P>0.05).After 4-week treatments,compared with the control group,the FMA-LE score was higher(P<0.05),and the peak blood flow velocities of PT and DP arteries on the affected side were larger in the observation group(P<0.05). Conclusion:EA combined with Western medication can significantly improve the motor function and blood flow velocity of the affected lower limb in patients with acute cerebral infarction accompanied by hemiplegia.
10.The crosstalk of Wnt/β-catenin signaling and p53 in acute kidney injury and chronic kidney disease
Wen-Hua MING ; Lin WEN ; Wen-Juan HU ; Rong-Fang QIAO ; Yang ZHOU ; Bo-Wei SU ; Ya-Nan BAO ; Ping GAO ; Zhi-Lin LUAN
Kidney Research and Clinical Practice 2024;43(6):724-738
Wnt/β-catenin is a signaling pathway associated with embryonic development, organ formation, cancer, and fibrosis. Its activation can repair kidney damage during acute kidney injury (AKI) and accelerate the occurrence of renal fibrosis after chronic kidney disease (CKD). Interestingly, p53 has also been found as a key modulator in AKI and CKD in recent years. Meantime, some studies have found crosstalk between Wnt/β-catenin signaling pathways and p53, but more evidence is required on whether they have synergistic effects in renal disease progression. This article reviews the role and therapeutic targets of Wnt/β-catenin and p53 in AKI and CKD and proposes for the first time that Wnt/β-catenin and p53 have a synergistic effect in the treatment of renal injury.

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