1.The effects and mechanisms of Gankyrin silencing on reversing the cisplatin resistance of human ovarian cancer SKOV3/DDP cell line
China Oncology 2014;(1):35-40
Background and purpose: Ovarian cancer is the common gynecological cancer, and the drug resistance of anti-tumor drug was one of major reasons for therapy failure, some studies considered that there is a closed relationship between Gankyrin and drug resistance. In this study, we investigated the effects and mechanisms of Gankyrin silencing on reversing the cisplatin resistance of ovarian cancer drug-resistant SKOV3/DDP cell line. Methods:The expression of Gankyrin in SKOV3 and SKOV3/DDP cells was measured by real-time PCR assay, MTS assay was employed to determine the effect of Gankyrin on SKOV3/DDP sensitivity to cisplatin, apoptosis rate and intracellular concentration of rhodamine-123 (Rh-123) were determined by lfow cytometry, the expression of multi-drugs resistant protein MDR1, Caspase-3/8, Survivin and Bcl-2 were determined by Western blot and real-time PCR. The phosphorylation of AKT and expression of p53, NF-κB and PTEN were analyzed by Western blot assay. Results:The expression of Gankyrin was increased in SKOV3/DDP cells, Gankyrin silencing was able to increase the cisplatin sensitivity of SKOV3/DDP. Before and after gene silencing, the reverse folds (RF) to cisplatin were 1.81 and 2.45, respectively, the intracellular levels of Rh-123 were 1.73 and 2.42 fold, the apoptosis rates were 2.23 and 4.23 fold,the expressions of MDR1, Survivin and Bcl-2 were downregulated, the mRNA expressions of MDR1 were 62.8%and 21.6%, the mRNA expressions of Survivin were 24.5%and 10.3%, the mRNA expressions of Bcl-2 were 47.5%and 18.4%, the levels of Caspase-3/8, p53 and PTEN were elevated, phosphorylation of AKT and expression of NF-kB were downregulated compared with control group. Conclusion:Gankyrin silencing was able to reverse the cisplatin resistance of SKOV3/DDP cells by inhibiting the drug eflfux and promoting cell apoptosis, the PTEN/AKT/NF-κB/p53 may be the key pathway.
2.Clinical therapeutic effect of Neurotropin combined with oxycodone hydrochloride for neuropathic cancer pain
Xianjiang CHENG ; Weishuai LIU ; Kun WANG
Chinese Journal of Clinical Oncology 2015;42(11):546-549
Objective:To investigate the clinical efficacy of Neurotropin combined with oxycodone hydrochloride in moderating the severe pain in neuropathic cancer pain (NCP) patients. Methods: NCP patients who received drug therapy with a visual analog scale (VAS) score of>4 were randomly divided into the placebo combined oxycodone group (group A) and Neurotropin combined oxy-codone group (group B). The VAS score, pain relief rate, frequency of pain outbreaks, average dose of oxycodone per day, and adverse drug reactions between the two groups were compared. Results:The VAS scores in groups A and B both had significant reduction after treatment (P<0.05), whereas the VAS score in group B after 14 days of treatment decreased more significantly than that in group A (P=0.03). The pain relief rate in group B patients 14 days after treatment was significantly higher than that in group A (P<0.001). The out-break pain in groups A and B 7 and 14 days after treatment significantly decreased, whereas the outbreak pain in group B was signifi-cantly lower than that in group A (P values were 0.07 and 0.07, respectively). The average dose of oxycodone per day in group B 14 days after treatment was lower than that in group A (P<0.001). Adverse reactions, such as nausea and vomiting, in group B were signifi-cantly less than those in group A (P<0.05). Conclusion:Neurotropin combined with oxycodone can effectively lower the NCP, average dose of oxycodone per day, and adverse reactions.
3.Efficacy of percutaneous vertebroplasty with radiotherapy for bone metastasis pain
Xianjiang CHENG ; Yuejuan SHAO ; Kun WANG
Chinese Journal of Clinical Oncology 2016;43(9):371-375
Objective:To compare efficacy of percutaneous vertebroplasty (PVP) with radiotherapy and radiotherapy alone for bone me-tastasis pain. Methods:A total of 247 bone metastasis patients with pain were analyzed. The radiotherapy group comprised 158 cases, whereas the combination group comprised 89 cases. We mainly observed the effect of pain treatment, behavioral states, and im-proved emotional condition. The side effects and complications were also investigated. Daily medicine consumption of background pain treatment was observed between the two groups. Analysis was done by SPSS 17.0 statistical software. Numerical variables were analyzed using t test and comparisons between groups used chi-square test. Results:The VAS scores of radiotherapy group decreased from 8.12±1.45 to 3.06±1.68 after treatment (P<0.05), and combination group VAS scores from 8.46±1.73 to 2.45±1.47 (P<0.05). The time to pain relief following PVP and radiotherapy were 1.63±0.81 and 8.56±2.87 days, respectively (P<0.001). The breakthrough pain frequency was 4.56 ± 1.98 times/day, which decreased to 1.57 ± 0.98 times/day after PVP (P<0.05). By contrast, the breakthrough pain frequency was 4.73±2.24 times/day before treatment, which decreased to 3.56±1.56 times/day after radiotherapy. No serious compli-cations were observed in the two groups. The depression and anxiety mood in the combination group improved after treatment. Daily medicine consumption in radiotherapy group increased after therapy. However, daily medicine consumption in combination group was reduced after therapy. Conclusion:PVP with radiotherapy can effectively relieve bone metastasis pain and improve patients' quality of life and it is worthy of promotion in clinical practice.
4.Preparation of TPGS-modified artesunate liposomes and their in vitro anti-tumor activity
Cheng HU ; Kun LIANG ; Rui AN ; Xinhong WANG ; Lisha YOU
Chinese Traditional Patent Medicine 2017;39(3):492-498
AIM To prepare D-α-tocopherol polyethylene glycol 1000 succinate (TPGS)-modified artesunate liposomes and to investigate the in vitro anti-tumor activity.METHODS The liposomes prepared by thin-film dispersion method were characterized by transmission electron microscopy and particle size analyzer,and the encapsulation efficiency was determined by ultrafiltration centrifugation.The liposomes' cytotoxicity to human hepatoma HepG2 cells was evaluated by MTT method.RESULTS The average particle size,PDI,Zeta potential,encapsulation efficiency,drug loading of the liposomes were 126.7 nm,0.182,-10.1 mV,78.8% and 18.38%,respectively.The liposomes displayed a significant inhibition on HepG2 cells with the IC50 value of 0.034 μmol/mL.CONCLUSION Compared with non-TPGS-modified artesunate liposomes,the TPGS-modified artesunate liposomes prepared by this method afford smaller vesicle size,better stability and higher encapsulation efficiency with stronger in vitro anti-tumor activity.
5.Cloning and Expression of a Bradyzoite-Specific Gene of Toxoplasma gondii and Immunoreactive Analysis on the Recombinant Antigen
Qiong WANG ; Kun WU ; Xiaoguang CHEN ; Li HAO ; Lu CHENG
Chinese Journal of Parasitology and Parasitic Diseases 1987;0(04):-
Objective To clone and express bradyzoite antigen 1(BAG1) gene of T. gondii,and analyze the immunoreactivity of the recombinant product. Methods The differentiation of T. gondii RH strain tachyzoites into bradyzoites was induced in vitro,and the coding sequence of BAG1 was amplified from bradyzoites by RT-PCR. The PCR product was analyzed by sequencing. The BAG1 coding sequence was further subcloned into the plasmid pET32a(+). The plasmid pET32a(+)-BAG1 was then transformed into BL21(DE3) to express after IPTG induction. The expression product was purified with Ni-NTA agarose and the purified BAG1 was further analyzed by Western blotting and ELISA. Results BAG1 cDNA was amplified from bradyzoites. After IPTG induction,BAG1 was expressed in a fusional form in E. coli. Western blotting showed that the purified recombinant protein could be specifically recognized by sera from mice chronically infected by T. gondii B36 strain. ELISA showed that the positive rate of T. gondii IgG antibodies of 350 human sera detected by the recombinant BAG1(17.4%) was higher than by recombinant SAG1 (12.6%)(P
6.Effects of interferon-γ on interleukin-10 and mononuclear macrophages in a mouse model of gallbladder cancer
Chunlin GE ; Tao SUN ; Ying CHENG ; Kun WANG
Chinese Journal of Digestive Surgery 2014;13(1):51-54
Objective To investigate the effects of interferon-γ (IFN-γ) on interleukin-10 (IL-10) and mononuclear macrophages in a mouse model of gallbladder cancer.Methods Mouse models of gallbladder cancer were constructed by inoculating the human gallbladder cancer cell line GBC-SD subcutaneously in 20 BALB/C mice,and then all the mice were randomly divided into the IFN-γ group and the control group (10 mice in each group).Murine recombinant IFN-γ (0.1 mL,1 × 105 kU/L,diluted with normal saline) was injected into the tumors in the IFN-γgroup,and normal saline was injected into the tumors in the control group.The expression of IL-10 was detected by ELISA,and the numbers of CD14 + cells (mononuclear macrophages),CD64 + cells (M1 macrophages) and CD206+ cells (M2 macrophages) were counted by the immunohistochemistry.All data were analyzed using the Student's t test.Results The mouse models of gallbladder cancer were successfully constructed 1 week later.Nine mice survived in the IFN-γ group,and 7 mice survived in the control group.The tumor weight was (518 ± 138)mg in the IFN-γ group and (669 ± 128)mg in the control group,with a significant difference between the 2 groups (t =2.240,P > 0.05).The volume of the tumor was (456 ± 172)mm3 in the IFN-γ group and (505 ± 146)mm3 in the control group,with no significant difference between the 2 groups (t =1.503,P > 0.05).The concentration of IL-10 was (58 ± 16) μg/g in the IFN-γgroup,which was significantly lower than (102 ± 45) μg/g in the control group (t =2.796,P < 0.05).The number of mononuclear macrophages was 81 ± 16 in the IFN-γ group,which was significantly greater than 50 ± 21 in the control group; the number of M1 macrophages was 66 ± 12 in the IFN-γ group,which was significantly greater than 9 ± 4 in the control group ; the number of M2 macrophages was 15 ± 4 in the IFN-γgroup,which was significantly lower than 40 ± 14 in the control group (t =3.214,13.127,6.914,P < 0.05).Conclusions IFN-γ could decrease the concentration of IL-10 in the tumor microenvironment,and it could induce the mononuclear macrophage to infiltrate into the stroma of the gallbladder cancer cells,and most of the monocytes and macrophages were differentiate to M1 macrophages.Gallbladder neoplasms; Interleukin-10; Interferon-γ; Mononuclear macrophages
7.Curative effectiveness of human chorionic gonadotropin treatment on hypospadias with micropenis
Chao CHEN ; Tiquan YANG ; Chunqiang DONG ; Qiang LIU ; Kun DONG ; Cheng SU ; Congjun WANG ; Cheng ZHANG
Chinese Journal of Applied Clinical Pediatrics 2017;32(11):821-824
Objective To evaluate the curative effectiveness and safety of human chorionic gonadotropin(HCG) treatment on children with microphallic hypospadias.Methods A total of 48 consecutive children with microphalic hypospadias were enrolled in the study,and the children were randomized into the experiment group(HCG treatment) and the control group with the research randomizer.The patients in experiment group were treated with HCG prior to surgery,and the control group did not received any hormone therapy preoperatively.All children in the experiment group and the control group underwent hypospadias repair by using transverse preputial island flap (Duckett technique) urethroplasty.Penile length,diameter of glans penis,bone age,serum testosterone level,and secondary effects were recorded before and after therapy in the experiment group.Postoperative complications were assessed with respect to fistulas,urethral strictures,diverticula,meatal stenosis,and glanular dehiscence in both groups.Results (1)Mean penile length and diameter of the experiment group increased significantly by (1.08±0.47) cm (t=-5.196,P<0.05) and (0.31±0.06) cm and there was a significant difference between before and after treatment (t=-5.080,P<0.05).(2)Urethrocutaneous fistulas were observed in 8 patients in the control group compared to 2 patients in the experiment group with a statistically significant difference (χ2=4.547,P<0.05).There was a significant difference between the overall reoperation rates of control group (9 patients) and the experiment group (3 patients,χ2=4.000,P<0.05).The penile tissue of the patients in the experiment group was soft and able to be easily separated and released during the operation and the flap had more blood supply.Conclusions Pretreatment with HCG therapy prior to hypospadias repair is beneficial to children with microphallic hypospadias.Significant penile growth was seen in the children treated with HCG and there was no obvious side effect.Moreover,pretreatment with HCG is beneficial to decrease the complications and reoperation rates of hypospadias repair which proves to be effective and safe.
8.Sarpogrelate hydrochloride in the treatment of chronic arterial ischemia of the lower extremities
Jun CHENG ; Xuehu WANG ; Le XIAO ; Jianming SUN ; Kun WANG ; Jianghua ZHENG ; Yu ZHAO ; De SHI
Chinese Journal of General Surgery 2012;(11):890-892
Objective To evaluate the clinical efficacy of oral Sarpogrelate hydrochloride in the treatment of chronic arterial ischemia of the lower extremities.Methods In this study 892 patients,who suffered from arteriosclerosis (ASO) or thromboangiitis obliterans ( TAO ) or diabetic foot ( DF ),with symptoms of intermittent claudication, sensation of cold, pain, ulcer, and without a history of vasotransplantation or bypass grafting or interventional therapy, were treated by taking Sarpogrelate hydrochloride tablets 100 mg tid for consecutive 8 weeks.The improvement rate of concomitant symptoms and the total effective rate of ASO, TAO, DF were evaluated.Drug adverse reaction were recorded.Results The improvement rate of intermittent claudication,sensation of cold,pain and ulcer were 96.9%,97.1%,89.0% and 86.9% respectively.The total effective rate for ASO,TAO,DF was 83.5%.A total of 81 cases (9.1%) reported mild side effects,including 7 patients with mild rash after 2- 5 days' medication,21 patients with mild nausea and 53 patients with stomach discomfort after 1 - 2 days' medication.Symptoms were managed conservatively without discontinuing taking sarpogrelate hydrochloride.Conclusions Sarpogrelate hydrochloride oral is a safe and effective therapy for chronic arterial ischemia diseases of the lower extremities.
9.Application study of malignant pancreas tumor treatment with embedment of ~(125)I by CT guide
Xiao-Kun HU ; Cheng-Bin YIN ; Shao-Kui WANG ; Yong-Kui WANG ; Jin JIN ;
Cancer Research and Clinic 2006;0(10):-
Objective To discuss the method,safety and effect of malignant pancreas tumor treated by CT guided percutaneous embedding of~(125)I. Methods 32 cases of malignant pancreas tumor with CT scan and contrast enhancement were retrospectively analysed.All the cases had been confirmed pathologically be- fore CT guided therapy.The number of~(125)I particle was 12~46,The distance between particles was 0.~1.2 cm. The number of puncture point was 1~2.The number of puncture direction was 2-5 times.Results Regarding 32 cases for 1 month,the size of the tumor reduced in 11 cases,no change in 20 cases,and increased in 1 case.As for 31 cases for 2 months,the size reduced in 16 cases,no change in 13 cases,and enlarged in 3 cases.Regarding 30 cases for 3 months,the size reduced in 18 cases,no change in 11 cases,and increased in 3 cases. Regarding 28 cases for 6 months, he size reduced in 10 cases, no change in 5 cases, and in- creased in 13 cases.Regarding 22 cases for 1 year,12 cases had been done the second therapy,the size of the tumor reduced in 16 cases,no change in 3 cases,and increased in 3 cases.Conclusion The size of pancreas tumors were reduced obviously,the symptoms were relieved after the treatment.The method turned out to be safe and accurate.
10.Expression of monocyte chemotactic protein-1 and its receptor in sudden coronary death.
Yuan-yuan KUANG ; Xia-xia CHEN ; Cang-cheng WANG ; Kun YE ; Ying WANG ; Yong-hua SHI
Journal of Forensic Medicine 2014;30(6):413-418
OBJECTIVE:
To investigate the expression of monocyte chemotactic protein-1 (MCP-1) and its receptor CC chemokine receptor-2 (CCR-2) in coronary atherosclerosis plaques between sidden coronary death (SCD) and non-SCD. Methods The expression levels of MCP-1 and CCR-2 in SCD group, coronary atherosclerosis group (non-SCD), control group (normal coronary artery) were detected by immunohistochemistry.
RESULTS:
Positive rates of MCP-1 among the three groups were 78%, 47%, and 0%, respectively, with significant expressing differences between each two groups (P<0.05). Positive rates of CCR-2 among three groups were 72%, 47%, and 0%, respectively, with significant expressing differences between the SCD group and coronary atherosclerosis group as well as between the SCD group and control group (P<0.05), but with no significant expressing difference between coronary atherosclerosis group and control group (P>0.05).
CONCLUSION
Overexpression of MCP-1 and CCR-2 in coronary atherosclerotic plaques is closely correlated with SCD.
Chemokine CCL2/metabolism*
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Coronary Artery Disease/pathology*
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Death, Sudden, Cardiac/pathology*
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Humans
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Immunohistochemistry
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Receptors, CCR2/metabolism*