1.miR-222 can inhibit the autophagy of renal cell carcinoma cells through down-regulating the expression of DDIT4
Xiaochen NI ; Zhihong ZHAO ; Yongliang MA ; Zongtao REN ; Bin LIU ; Aili ZHANG
China Oncology 2015;(3):161-166
Background and purpose:MicroRNA (miRNA, miR) plays an important regulatory role in cancer. miR-222 is reported to be up-regulated in various tumors, but its role in renal cell carcinoma (RCC) remains unclear. In this study, we detected the expression of miR-222 in both RCC and adjacent tissue samples. The aim of this study was to investigate the role of miR-222 in RCC. Methods:The expression levels of miR-222 in RCC tissue samples were quantified by quantitative real-time polymerase chain reaction (qRT-PCR). DDIT4 and LC3-Ⅱ protein expressions were determined by Western blot. Dual luciferase assay was performed to verify the target of miR-222. EGFP-LC3 microscopy assay was performed to assess autophagy. Results:Results from qRT-PCR showed that the expression of miR-222 was up-regulated in RCC tissues. Knockdown of miR-222 with speciifc antagomiR decreased the cell viability of 786-O cells, whereas overexpression of miR-222 increased the cell viability (P<0.01). The levels of DDIT4 were up-regulated in 786-O cells transfected with miR-222 antagomiR, whereas overexpression of miR-222 induced the down-regulation of DDIT4 expression. Data from dual luciferase assay indicated that miR-222 directly targeted the expression of DDIT4. Consistently, the expression of DDIT4 in RCC tissues was down-regulated compared with adjacent tissues. Knockdown of miR-222 in 786-O cells induced a signiifcant increase of autophagosome formation and LC3 lipidation.These results supported that miR-222 could inhibit autophagy in RCC cells, which may affect the clinical characteristcs of RCC. Conclusion: miR-222 is up-regulated in RCC and can inhibit the autophagy of RCC cells through down-regulating the expression of DDIT4.
2.Hinokitiol induces clear cell renal cancer 786-O cell apoptosis via autophagy induction
Xiaochen NI ; Zhihong ZHAO ; Yongliang MA ; Zongtao REN ; Bin LIU ; Bo FAN ; Shufei WEI ; Aili ZHANG
Chinese Journal of Clinical Oncology 2015;(1):43-46
Objective: To investigate the effects of hinokitiol on the proliferative inhibition and apoptosis induction in human clear cell renal cancer 786-O cells. Methods:CCK-8 assays were performed to analyze the effects of hinokitiol on the proliferation of 786-O cells. The apoptosis rate was determined by flow cytometry. EGFP-LC3 microscopy assays were performed to assess the autoph-agy flux. Cleaved Caspase-3, LC3, and P62 were detected by Western blot. Results: Hinokitiol could inhibit the proliferation of the 786-O cells and could induce cell apoptosis via Caspase pathway. Hinokitiol induced the autophagy of 786-O cells, increased LC3 ex-pression, and downregulated P62 expression. Conclusion: Hinokitiol can inhibit the proliferation of 786-O cells and can induce cell apoptosis via autophagy induction.
3.DACT2 gene promoter area methylation status and mRNA expression in renal cell carcinoma
Bo FAN ; Pan QI ; Aili ZHANG ; Zhihong ZHAO ; Xiaochen NI ; Bin LIU ; Yongliang MA ; Zongtao REN
Chongqing Medicine 2017;46(21):2895-2897,2901
Objective To explore the role of the DACT2 gene in the occurrence and development of renal cell carcinoma(RCC).Methods The samples of RCC tissues and corresponding tumor-adjacent tissues after radical operation and normal kidney tissues were collected.The methylation specific PCR (MSP) and real time fluorescence reverse transcriptase-PCR (RT-PCR) methods were adopted to detect the methylation status and mRNA expression of DACT2.The streptavidin-peroxidase (SP) method labeled by immunohistochemistry peroxidase was used to examine the expression of β-catenin protein.Then the relationship between DACT2 gene methylation status and mRNA expression with the clinicopathologic characteristics was analyzed.The relationship between DACT2 gene methylation with mRNA and β-catenin expression was analysed,as well.Results The DACT2 mRNA relative expression level in RCC tissues was 0.427±0.025,which was significantly lower than (0.801±0.047) in tumor-adjacent tissues and (0.872±0.022) in normal tissue,the positive rate of DACT2 gene methylation in RCC tissues was 45.76%,which was significantly higher than 6.78% in tumor-adjacent tissues and 5.08% in normal tissues,the difference was statistically significant (P<0.05),while the difference between tumor-adjacent tissues and normal tissues had no statistical significance (P>0.05).The DACT2 gene mRNA expression level in RCC tissues and promoter area methylation occurrence rate had no obvious correlation with the clinical data such as patients age,gender,tumor size,clinical stage and Fuhrman grade (P>0.05).The DACT2 gene mRNA relative level in the methylation group was lower than that in the non-methylation group,the difference was statistically significant (P<0.05).The expression rate of β-catenin protein in cytoplasma in RCC tissues was higher than that in the tumor-adjacent tissues and normal tissues,the difference was statistically significant (P<0.05),moreover,DACT2 gen methylation had a positive correlation with β-catenin protein expression (r=0.324,P=0.012).Conclusion The decrease of DACT2 gene promoter area methylation and mRNA relative expression level may participate in the RCC occurrence,but has no relationship with RCC clinical progression.Methylation occurred in DACT2 gene promoter area may be one of reasons causing mRNA relative expression decrase.DACT2 gene methylation occurrence in RCC tissue might be related to the high expression of β-catenin.
4.The effect study of bladder perfusion with sodium hyaluronate combined with ultrashort wave in treatment of glandular cystitis
Yongliang NI ; Haixin WANG ; Qiang ZHOU ; Benkang SHI
Chinese Journal of Urology 2017;38(z1):34-39
Objective To evaluate the efficacy of bladder perfusion with sodium hyaluronate combined with ultrashort wave irradiation in bladder region in treatment of glandular cystitis.Methods 22 patients diagnosed as typical glandular cystitis were selected from our hospital between March 2013 and March 2013. Those patients were given bladder perfusion with sodium hyaluronate combined with ultrashort wave irradiation in bladder region according to the designed course. The lesion areas were taked photos and bited samples for microscopic observation and HE staining.The evaluation indicators include pelvic pain and urgency/frequency symptom scores,cystitis histologic scores ,infiltrating lymphocyte counts, Brunn nest or cystica counts and residual urine volume were measured in the self-control expertiment before treatment and after. Results Over the course of treatment, above mentioned indicators had significant improvement compared with original data among those patients (P<0.01).Only 1 patient appeared itchy skin two times after bladder perfusion with sodium hyaluronate, others without serious complications. Conclusions The method that bladder perfusion with sodium hyaluronate combined with ultrashort wave irradiation has clear effect without serious complications.This method provides a new train of thought for glandular cystitis and opens up a new space.
5.Coordinated Hospital-Home Fecal Microbiota Transplantation via Percutaneous Endoscopic Cecostomy for Recurrent Steroid-Dependent Ulcerative Colitis.
Xiaodong NI ; Shengxian FAN ; Yongliang ZHANG ; Zhiming WANG ; Lan DING ; Yousheng LI ; Jieshou LI
Gut and Liver 2016;10(6):975-980
Since its introduction as an alternative intestinal microbiota alteration approach, fecal microbiota transplantation (FMT) has been increasingly used as a treatment of choice for patients with ulcerative colitis (UC), but no reports exist regarding FMT via percutaneous endoscopic cecostomy (PEC). This report describes the case of a 24-year-old man with a 7-year history of recurrent, steroid-dependent UC. He received FMT via PEC once per day for 1 month in the hospital. After the remission of gastrointestinal symptoms, he was discharged from the hospital and continued FMT via PEC twice per week for 3 months at home. The frequency of stools decreased, and the characteristics of stools improved soon thereafter. Enteral nutrition was regained after 1 week, and an oral diet was begun 1 month later. Two months after the FMT end point, the patient resumed a normal diet, with formed soft stools once per day. The follow-up colonoscopy showed normal mucus membranes; then, the PEC set was removed. On the subsequent 12 months follow-up, the patient resumed orthobiosis without any gastrointestinal discomfort and returned to work. This case emphasizes that FMT via PEC can not only induce remission but also shorten the duration of hospitalization and reduce the medical costs; therefore, this approach should be considered an alternative option for patients with UC.
Cecostomy*
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Colitis, Ulcerative*
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Colonoscopy
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Diet
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Enteral Nutrition
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Fecal Microbiota Transplantation*
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Follow-Up Studies
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Gastrointestinal Microbiome
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Hospitalization
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Humans
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Membranes
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Mucus
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Ulcer*
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Young Adult
6.Reproducibility of a portable spirometer based on differential pressure sensor.
Kun ZOU ; Chaomin NI ; Zhijun HE ; Yongliang ZHANG ; Zuchang MA ; Jianxia SONG ; Ke ZHANG
Chinese Journal of Medical Instrumentation 2013;37(1):30-48
OBJECTIVEAssessing the reproducibility of a portable spirometer, including reproducibility of inter-observer and day-today.
METHODSLung ventilation function was performed in 22 healthy volunteers by two observers on the same day and repeated by the first observer after 24h.
RESULTSThe inter-observer and day-to-day intra-class correlation coefficients are all higher than 0.75. There are no significant difference between each other. Bland-Altman chart shows good limits of agreement between inter-observer and day-to-day, only scattered data are outside of the limits of agreement.
CONCLUSIONSThe portable spirometer shows good inter-observer and day-to-day reproducibility, and can be used for testing lung function in clinical.
Adult ; Female ; Humans ; Male ; Middle Aged ; Monitoring, Ambulatory ; instrumentation ; Observer Variation ; Reproducibility of Results ; Spirometry ; instrumentation
7.The distribution characteristics of urine flora in patients with ureteral stent tube crusting
Yongliang NI ; Wei WEI ; Xiangtao WANG ; Xiaolu SUN ; Zhongxian HUANG ; Bo WANG ; Mingjie LI ; Deqi JIANG ; Yunwei LI ; Qiang WEI ; Xia LIU ; Benkang SHI
Chinese Journal of Urology 2020;41(4):262-266
Objective:To investigate the distribution characteristics of bacteria in urine of patients with ureteral stent crusting.Methods:Thirty-five patients who underwent ureteral stent placement at the Shandong Provincial Third Hospital, Shandong University Qilu Hospital, Jinan Central Hospital, and Jinan Jigang Hospital were selected from October, 2018 to March, 2019(the clinical study registration number is ChiCTR1800020025). The inclusion criteria were patients who had the stent intubated for 4 weeks after ureteroscopic lithotripsy, aged between 18 and 65 years. Exclusion criteria were patients with positive urine bacterial culture, severe gross hematuria, recent oral antibiotics, and patients with significant residual stones. This clinical study uses a cross-sectional study method, and those patients were divided into crusting group (n=23) and non-crusting group (n=12) according to the presence or absence of stent crusting. On the day of extubation, urine of the patients was collected for bacterial 16s DNA detection. The distribution characteristics of bacteria in urine of the two groups were analyzed using UPARSE, UCHIME and RDP calssifier. The total number of bacteria species, bacterial abundance and bacterial species with large-scale abundance in urine of the two groups were determined. The quantity of bacteria species and bacterial abundance in the urine between the two groups were compared, and the bacterial species with large-scale abundance in urine of the patients with stent crusting were identified.Results:There were no significant differences in general information such as age, body mass index, gender, affected side, type of stent tube, and stone composition between the two groups. Using 16s DNA sequencing to detect the bacteria in the urine of the two groups revealed that the number of bacterial species with abundance >1% was 11, and the number of bacterial species with abundance >0.01% was 74 in the crusting group. In the non-crusting group, the number of bacterial species with abundance >1% and >0.01% was 7 and 11, respectively. Compared with the non-crusting group, the number of bacterial species with abundance >1% in the crusting group was significantly larger ( t=5.12, P=0.000). In the crusting group, bacterial species with the top three abundance were g_Lactobacillus (23.1%), g_Bacteroides (18.8%) and g_norank_Bacteroides (17.1%). In the non-crusting group, bacterial species with the top three abundance were g_Escherichia-Shigella (32.2%), g_Enterococcus (24.9%) and g_Pseudomonas (18.2%). The three bacteria with the greatest difference between the two groups were g_ Lactobacillus ( P=0.010), g_Bacteroides ( P=0.004) and g_norank_Bacteroides ( P=0.004), respectively. Conclusion:The species and quantity of bacteria in the urine of patients with stent crusting are both significantly larger than those of patients without stent crusting. Bacteroides with larger-scale abundance in the urine of patients with stent crusting may promote the deposition of crystals on the stent wall through its structure, function and urease positive characteristics.