1.Professor WANG Zimin's Experience in Treating IgA Nephropathy by Stages and Types
Kunpeng FAN ; Haiyan XING ; Yabin GAO
Journal of Zhejiang Chinese Medical University 2024;48(5):572-575
[Objective]To summarize Professor WANG Zimin's experience in the differentiation and treatment of IgA nephropathy by stages and types.[Methods]Through learning from teachers,collecting medical records,summarizing and exploring Professor WANG's experience in treating IgA nephropathy,and attaching a medical record.[Results]Professor WANG believes that IgA nephropathy belongs to a disease with a combination of deficiency and excess,mainly located in the kidney and closely related to the liver,spleen and lungs.The etiology is divided into internal and external causes,which are combined to form the disease.He also believes that the core pathogenesis of this disease is"dampness,heat,blood stasis and deficiency".The disease is divided into acute attack stage and chronic persistent stage.The acute attack stage is divided into wind heat attacking the lung type,heart fire hyperactivity type,dampness turbidity invading the spleen type and lower Jiao dampness heat type;chronic persistent stage can be divided into Yin deficiency and fire excess type,Qi and Yin deficiency type,spleen and kidney deficiency type,and blood stasis and toxin obstruction type.Professor WANG treats different syndrome types with Yinqiao Powder,Daochi Powder,Xiaoji Yinzi,Zhibai Dihuang Pill and Erzhi Pill,Huangqi Dihuang Decoction,Liujunzi Decoction and Yougui Pill,self-made Huanghuai Wendan Decoction and Xuefu Zhuyu Decoction,and other modified treatments.Professor WANG points out that the treatment of this disease should be based on the principles of nourishing the body,dispelling pathogenic factors,and combining traditional Chinese and western medicine,and advocates for a combination of traditional Chinese and western medicine,and he is good at using drug pair for precise treatment.In the attached medical record,the initial diagnosis was identified as Yin deficiency and excessive fire type.The treatment was to nourish Yin and reduce fire,clear heat and cool blood,modified Zhibai Dihuang Pill and Erzhi Pill were used,and then added or subtracted the herbs according to the symptoms,resulting in excellent therapeutic effects.[Conclusion]Professor WANG has rich experience in staging and typing the treatment of IgA nephropathy,and proposes that the core pathogenesis of this disease is"dampness,heat,stasis and deficiency",which has high guiding significance for the treatment of this disease.
2.Analysis of surgical situations and prognosis of pancreaticoduodenectomy in Jiangsu province (a report of 2 886 cases)
Zipeng LU ; Xin GAO ; Hao CHENG ; Ning WANG ; Kai ZHANG ; Jie YIN ; Lingdi YIN ; Youting LIN ; Xinrui ZHU ; Dongzhi WANG ; Hongqin MA ; Tongtai LIU ; Yongzi XU ; Daojun ZHU ; Yabin YU ; Yang YANG ; Fei LIU ; Chao PAN ; Jincao TANG ; Minjie HU ; Zhiyuan HUA ; Fuming XUAN ; Leizhou XIA ; Dong QIAN ; Yong WANG ; Susu WANG ; Wentao GAO ; Yudong QIU ; Dongming ZHU ; Yi MIAO ; Kuirong JIANG
Chinese Journal of Digestive Surgery 2024;23(5):685-693
Objective:To investigate the surgical situations and perioperative outcome of pancreaticoduodenectomy in Jiangsu Province and the influencing factors for postoperative 90-day mortality.Methods:The retrospective case-control study was conducted. The clinicopathological data of 2 886 patients who underwent pancreaticoduodenectomy in 21 large tertiary hospitals of Jiangsu Quality Control Center for Pancreatic Diseases, including The First Affiliated Hospital of Nanjing Medical University, from March 2021 to December 2022 were collected. There were 1 732 males and 1 154 females, aged 65(57,71)years. Under the framework of the Jiangsu Provincial Pancreatic Disease Quality Control Project, the Jiangsu Quality Control Center for Pancreatic Diseases adopted a multi-center registration research method to establish a provincial electronic database for pancrea-ticoduodenectomy. Observation indicators: (1) clinical characteristics; (2) intraoperative and post-operative conditions; (3) influencing factors for 90-day mortality after pancreaticoduodenectomy. Measurement data with skewed distribution were represented as M( Q1, Q3) or M(IQR), and comparison between groups was conducted using the Mann-Whitney U test. Count data were expressed as absolute numbers or constituent ratio, and comparison between groups was conducted using the chi-square test, continuity correction chi-square test and Fisher exact probability. Maximal Youden index method was used to determine the cutoff value of continuous variables. Univariate analysis was performed using the corresponding statistical methods based on data types. Multivariate analysis was performed using the Logistic multiple regression model. Results:(1) Clinical characteristics. Of the 2 886 patients who underwent pancreaticoduodenectomy, there were 1 175 and 1 711 cases in 2021 and 2022, respectively. Of the 21 hospitals, 8 hospitals had an average annual surgical volume of <36 cases for pancreaticoduodenectomy, 10 hospitals had an average annual surgical volume of 36-119 cases, and 3 hospitals had an average annual surgical volume of ≥120 cases. There were 2 584 cases performed pancreaticoduodenectomy in thirteen hospitals with an average annual surgical volume of ≥36 cases, accounting for 89.536%(2 584/2 886)of the total cases. There were 1 357 cases performed pancrea-ticoduodenectomy in three hospitals with an average annual surgical volume of ≥120 cases, accounting for 47.020%(1 357/2 886) of the total cases. (2) Intraoperative and postoperative conditions. Of the 2 886 patients, the surgical approach was open surgery in 2 397 cases, minimally invasive surgery in 488 cases, and it is unknown in 1 case. The pylorus was preserved in 871 cases, not preserved in 1 952 cases, and it is unknown in 63 cases. Combined organ resection was performed in 305 cases (including vascular resection in 209 cases), not combined organ resection in 2 579 cases, and it is unknown in 2 cases. The operation time of 2 885 patients was 290(115)minutes, the volume of intra-operative blood loss of 2 882 patients was 240(250)mL, and the intraoperative blood transfusion rate of 2 880 patients was 27.153%(782/2 880). Of the 2 886 patients, the invasive treatment rate was 11.342%(327/2 883), the unplanned Intensive Care Unit (ICU) treatment rate was 3.087%(89/2 883), the reoperation rate was 1.590%(45/2 830), the duration of postoperative hospital stay was 17(11)days, the hospitalization mortality rate was 0.798%(23/2 882), and the failure rate of rescue data in 2 083 cases with severe complications was 6.529%(19/291). There were 2 477 patients receiving postoperative 90-day follow-up, with the 90-day mortality of 2.705%(67/2477). The total incidence rate of complication in 2 886 patients was 58.997%(1 423/2 412). The incidence rate of severe complication was 13.970%(291/2 083). The comprehensive complication index was 8.7(22.6) in 2 078 patients. (3) Influencing factors for 90-day mortality after pancreaticoduodenectomy. Results of multivariate analysis showed that age ≥ 70 years, postoperative invasive treatment, and unplanned ICU treatment were independent risk factors for 90-day mortality after pancreaticoduodenectomy ( odds ratio=2.403, 2.609, 16.141, 95% confidence interval as 1.281-4.510, 1.298-5.244, 7.119-36.596, P<0.05). Average annual surgical volume ≥36 cases in the hospital was an independent protective factor for 90-day mortality after pancreaticoduodenectomy ( odds ratio=0.368, 95% confidence interval as 0.168-0.808, P<0.05). Conclusions:Pancreaticoduodenectomy in Jiangsu Province is highly con-centrated in some hospitals, with a high incidence of postoperative complications, and the risk of postoperative 90-day mortality is significant higher than that of hospitallization mortality. Age ≥ 70 years, postoperative invasive treatment, and unplanned ICU treatment are independent risk factors for 90-day motality after pancreaticoduodenectomy, and average annual surgical volume ≥36 cases in the hospital is an independent protective factor.
3.Application of optical surface monitoring system guided volumetric modulated arc therapy in total body irradiation
Zhuangling LI ; Heli ZHONG ; Yan GAO ; Longxing LI ; Yabin SHI ; Xiaonian DENG ; Xin FU ; Ding ZHANG ; Fang ZHENG ; Hongtao CHEN ; Weisi CHEN
Chinese Journal of Medical Physics 2024;41(9):1070-1077
Objective To establish a novel clinical application process of the optical surface monitoring system(OSMS)guided volumetric modulated arc therapy(VMAT)for total body irradiation(TBI),and to assess the accuracy and effectiveness of OSMS in inter-fractional auxiliary positioning before radiotherapy and real-time monitoring of intra-fractional motion during radiotherapy.Methods A retrospective analysis was conducted on 15 leukemia patients who underwent OSMS-guided VMAT-TBI before hematopoietic stem cell transplantation.CT simulation positioning was performed,and the whole-body image data which were collected in head-first supine position(HFS)and feet-first supine position(FFS)were transmitted to the treatment planning system for image registration,multicenter VMAT planning and dose verification.The prescription dose was 800 cGy in 4 fractions twice daily.OSMS was used to assist positioning before delivery,and CBCT was used for position verification.During treatment,OSMS was used for monitoring.The intra-fractional error monitored by OSMS in real time was obtained by analyzing the offline log files.Results The mean dose and coverage of the target area in HFS plan were(905.4±19.0)cGy and 93.0%±2.8%.The mean doses to lung and kidney were(603.7±55.7)cGy and(600.4±49.6)cGy,respectively,and the maximum dose to the lens was(393.9±58.9)cGy.The mean dose and coverage of the target area in FFS plan were(888.5±58.9)cGy and 94.0%±3.2%;and the maximum dose at the junction was(1148.9±72.9)cGy.Fractional treatment delivery time was(75.1±15.1)min.OSMS-assisted positioning was carried out before delivery,and the total deviations of CBCT three-dimensional vector in translational and rotation directions were(2.71±1.96)mm and 0.91°±0.90°,respectively.The three-dimensional vector deviation of the intra-fractional motion amplitude in translational direction monitored by OSMS during the treatment was(1.95±1.88)mm,of which the deviation within 1 mm accounted for 57.5%,79.7%and 62.1%in longitudinal,lateral and vertical directions,respectively.The three-dimensional vector deviation in rotation direction was 0.76°±0.72°,of which the deviation within 1°accounted for 93.1%,85.7%and 94.3%in rotation,pitch and roll directions,respectively.Conclusion VMAT simplifies TBI process,while improving target coverage and organs-at-risk sparing.The use of OSMS can reduce positioning errors,especially rotation errors.In order to ensure the accurate implementation of TBI and the safety of patients,it is necessary to use OSMS for auxiliary positioning and intra-fractional position monitoring.
4.Optimization of immobilization and positioning methods in hypofractionated stereotactic radiotherapy for brain metastases
Longxing LI ; Yan GAO ; Hongtao CHEN ; Zihuang LI ; Yabin SHI
Chinese Journal of Radiological Medicine and Protection 2024;44(7):587-593
Objective:To compare translational and rotational setup errors between immobilization with open masks combined with positioning with the optical surface monitoring system (OSMS) and immobilization with full masks combined with positioning with laser lights and mask markers, find the advantages of open masks combined with OSMS in hypofractionated stereotactic radiotherapy (HSRT) for brain metastases, and calculate planning target volume (PTV) expansions with different immobilization and positioning method for patients with brain metastases.Methods:The setup data of 55 patients with brain metastases who received HSRT were analyzed retrospectively. According to immobilization and positioning method, the patients were divided into group A (OSMS + open masks), group B1 (full head-neck-shoulders masks + polyurethane foam cushions), and group B2 (full head-neck-shoulders masks + standard headrests). Positioning was directed by automatic couch motion in OSMS in group A and by laser lights and mask markers in groups B1 and B2. Cone beam computed tomography (CBCT) scans were registered using the bone registration method to obtain setup errors in six directions ( x, y, z, roll, pitch, yaw). PTV expansions were calculated according to the van Herk formula. Results:A total of 288 sets of CBCT registration data were acquired. Among three groups, group A showed the smallest mean setup errors and 3D vector error, which were (0.47±0.33) mm, (0.49±0.31) mm, (0.44±0.31) mm, (0.42±0.32)°, (0.48±0.31)°, (0.42±0.22)°, and (0.90±0.39) mm, respectively. Group A differed significantly from group B1 and group B2 in the errors at all directions ( P < 0.05) except for the yaw direction compared with group B1 ( P > 0.05). Group A had no setup error ≥2 mm in translational directions or ≥2° in rotational directions. Group B1 showed significantly smaller setup errors in the y, z, and yaw directions and 3D vector error than group B2 ( P < 0.05). In group A, PTV expansions in three directions ( x, y, and z) were 1.32, 1.19, and 1.22 mm, respectively, which were smaller than those of the other two groups. Conclusions:In HSRT for patient with brain metastases, compared with full head-neck-shoulders masks combined with laser lights and mask markers, open masks combined with OSMS can significantly improve setup precision in six directions and reduce repeated setup and PTV expansions, which shows promise for clinical application.
5.Effect of air-oxygenated normothermic mechanical perfusion on rat kidney recovery following ischemic injury from donation after cardiac death
Zhu CHEN ; Kaikai CHI ; Peng LU ; Yabin GAO ; Qing ZHANG ; Kang YANG ; Zheng WANG
Chinese Journal of Organ Transplantation 2022;43(2):95-101
Objective:To develop an ex vivo normothermic mechanical perfusion(NMP)and compare the effect of air-oxygenated NMP versus oxygen-oxygenated NMP on reducing renal injury from donor after cardiac death(DCD).Methods:All kidneys from DCD rats were subjected to 30 min in situ warm ischemia after cardiac attest.And harvested kidneys were stored for 8h under static cold preservation after NMP for 2h.In experimental groups, kidneys were subjected to either air-oxygenated NMP(group A, n=6)or oxygen-oxygenated NMP(group O, n=6). Sham operation(group C, n=6)and DCD kidneys under static cold preservation without NMP(group SCS, n=6)were employed as controls.The evaluation parameters included creatinine(Cr), aspartate amino transferase(AST)and lactate dehydrogenase(LDH)in perfusate, pathological changes by hematoxylin-eosin(HE)staining, histological criteria, expressions of myeloperoxidase and intercellular adhesion molecular-1(ICAM-1)by immunohistochemistry and Western blot, tumor necrosis factor-alpha(TNF-α)and interleukin-6(IL-6)by enzyme-linked immunoadsorbent assay and level of malondialdehyde(MDA)by thiobarbital method and activity of superoxide dismutase(SOD)by WST-8 in renal tissues.Differences between two groups were analyzed by two-tailed unpaired Student's test and differences among more than two groups by one-way ANOVA.Results:Renal arterial oxygen tensions in NMP perfusate were(576.3±68.2)mmHg with oxygen-oxygenation and(137.0±39.1)mmHg with air-oxygenation.There was significant difference( P<0.05). The pathological injury scores in groups SCS, O and A by HE staining were(7.0±0.1), (5.0±0.9)and(2.5±0.5); injury scores and the expressions of renal proximal tubular epithelial cell vacuolar degeneration in groups O and A were lower than those in group SCS( P<0.05)and injury score in group A was lower than group O( P<0.05). In perfusate, the levels of △Cr, △AST and △LDH in groups O and A were(43.9±52.8)μmol/L and(12.6±3.5)μmol/L, (532.3±52.8)U/L and(49.1±50.4)U/L and(9998.0±2014.4)U/L and(1477.0±810.4)U/L.There were significant differences( P<0.05). In perfused kidneys, the MDA level and SOD activity in groups O and A were(0.192±0.018)mmol/g, (0.162±0.023)mmol/g, (0.6±0.3)×10 3 U/g, (1.7±0.4)×10 3 U/g; TNF-α and IL-6 levels in groups O and A were(124.376±19.635)and(89.331±13.123)ng/g, and(4.038±1.026)×10 3 and(1.774±0.518)×10 3 ng/g.After air-oxygenated NMP, lower renal damage indices were characterized by a lower MDA level and a higher SOD activity, the lower levels of TNF-α and IL-6 and the lower expressions of MPO and ICAM-1 than those in oxygen-oxygenated NMP( P<0.05). Conclusions:NMP with air-oxygenation mimics renal perfusion under physiological conditions and decreases oxidative stress and inflammation injury.It may confer a better retrieval in DCD kidney against warm ischemia injury.
6.Meta-analysis of dosimetric comparison between volumetric-modulated arc therapy and intensity-modulated radiotherapy for breast cancer after modified radical mastectomy
Yan GAO ; Heli ZHONG ; Zhuangling LI ; Guangwei ZHANG ; Longxing LI ; Yabin SHI ; Xianming LI
Chinese Journal of Radiation Oncology 2021;30(11):1159-1166
Objective:To compare the dosimetric differences between volumetric-modulated arc therapy (VMAT) and intensity-modulated radiation therapy (IMRT) on planning target volume (PTV) and organ-at-risk (OAR) for breast cancer after modified radical mastectomy, aiming to provide evidence-based reference for clinical practice.Methods:According to strict inclusion and exclusion criteria, literature search was performed in PubMed, Cochrane Library, FMRS, CNKI, Wanfang Data and VIP full text databases from the inception of databases up to March 2020. The controlled clinical trials of dosimetric comparison between VMAT and IMRT for breast cancer following modified radical mastectomy were selected. The meta-analysis was performed using Stata14 software.Results:The meta-analysis included 281 patients from 13 observational studies. Compared with IMRT, VMAT significantly increased the PTV dose coverage D 95%( P<0.001) and significantly improved the PTV homogeneity index (HI, P<0.001) and conformity index (CI, P=0.004). Compared with IMRT, VMAT more effectively decreased the ipsilateral lung V 20Gy (WMD=1.332, P=0.027) and contralateral lung V 10Gy ( P=0.003). There were no significant differences in theD mean, V 5Gy, V 10Gy and V 30Gy of the ipsilateral lung, D mean and V 5Gy of the contralateral lung, D mean, V 10Gy and V 30Gy of the heart between VMAT and IMRT (all P>0.05). Compared with VMAT, IMRT reduced the cardiac V 5Gy ( P=0.001). However, sensitivity analysis of included literature on cardiac V 5Gy showed that the P value was reversed, indicating that the stability of the results was poor. VMAT significantly shortened the delivery time ( P<0.001) and the number of monitor units ( P<0.001) compared to IMRT. Conclusion:Compared with IMRT, VMAT can achieves superior target dose coverage, HI and CI, better protection for the ipsilateral and contralateral lung, fewer monitor units and shorter delivery time.
7.Cardiac functional alterations and its risk factors in elderly patients with obstructive sleep apnea syndrome free of cardiovascular disease.
Yinghui GAO ; Yongfei WEN ; Xiaoshun QIAN ; Libo ZHAO ; Hu XU ; Weihao XU ; Xiaoxuan KONG ; Hebin CHE ; Yabin WANG ; Lin LIU
Journal of Southern Medical University 2020;40(11):1587-1592
OBJECTIVE:
To evaluate the changes of cardiac structure and function and their risk factors in elderly patients with obstructive sleep apnea syndrome (OSA) without cardiovascular complications.
METHODS:
Eighty-two elderly OSA patients without cardiovascular disease admitted between January, 2015 and October, 2016 were enrolled in this study. According to their apnea-hypopnea index (AHI, calculated as the average number of episodes of apnoea and hypopnoea per hour of sleep), the patients were divided into mild OSA group (AHI < 15) and moderate to severe OSA group (AHI ≥ 15). The demographic data and the general clinical data were recorded and fasting blood samples were collected from the patients on the next morning following polysomnographic monitoring for blood cell analysis and biochemical examination. Echocardiography was performed within one week after overnight polysomnography, and the cardiac structure, cardiac function and biochemical indexes were compared between the two groups.
RESULTS:
Compared with those with mild OSA group, the patients with moderate to severe OSA had significantly higher hematocrit (0.22±0.08
CONCLUSIONS
Cardiac diastolic function impairment may occur in elderly patients with moderate or severe OSA who do not have hypertension or other cardiovascular diseases, and the severity of the impairment is positively correlated with AHI.
Aged
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Cardiovascular Diseases/etiology*
;
Humans
;
Risk Factors
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Severity of Illness Index
;
Sleep Apnea, Obstructive/complications*
;
Stroke Volume
;
Ventricular Dysfunction, Left
;
Ventricular Function, Left
8.Correlation between severity of obstructive sleep apnea syndrome and red cell distribution width in elderly patients.
Shuping WU ; Yinghui GAO ; Xiaoshun QIAN ; Libo ZHAO ; Hu XU ; Weihao XU ; Xiaoxuan KONG ; Yang YANG ; Hebin CHE ; YaBin WANG ; Xina YUAN ; Lin LIU
Journal of Southern Medical University 2020;40(5):703-707
OBJECTIVE:
To investigate the correlation between the severity of obstructive sleep apnea syndrome (OSAS) and red cell distribution width (RDW) in elderly patients.
METHODS:
A cross-sectional study was conducted among 311 elderly patients diagnosed with OSAS in the snoring clinic between January, 2015 and October, 2016 and 120 healthy controls without OSAS from physical examination populations in the General Hospital of PLA. The subjects were divided into control group with apnea-hypopnea index (AHI) <5 (=120), mild OSAS group (AHI of 5.0-14.9; =90), moderate OSAS group (AHI of 15.0-29.9; =113) and severe OSAS group (AHI ≥ 30; =108). The clinical characteristics and the results of polysomnography, routine blood tests and biochemical tests of the subjects were collected. Multiple linear regression analysis was used to examine the correlation between OSAS severity and RDW.
RESULTS:
The levels of RDW and triglyceride were significantly higher in severe OSAS group than in the other groups ( < 0.01). The levels of fasting blood glucose and body mass index were significantly higher in severe and moderate OSAS groups than in mild OSAS group and control group ( < 0.05 or < 0.01). Multiple linear regression analysis showed that AHI was positively correlated with body mass index (β=0.111, =0.032) and RDW (β=0.106, =0.029). The area under ROC curve of RDW for predicting the severity of OSAS was 0.687 (=0.0001).
CONCLUSIONS
The RDW increases as OSAS worsens and may serve as a potential marker for evaluating the severity of OSAS.
Aged
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Cross-Sectional Studies
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Erythrocyte Indices
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Humans
;
Polysomnography
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Severity of Illness Index
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Sleep Apnea, Obstructive
9.Mechanism of lncRNA-SRLR induced invasion and metastasis in U2OS osteosarcoma cells
Fei CAO ; Xiaohong KANG ; Dafeng WANG ; Long MA ; Xiangjun CAO ; Ying WANG ; Yuanyuan GAO ; Zhanhui MIAO ; Haibin DENG ; Yabin GONG
Chinese Journal of Oncology 2020;42(12):1007-1013
Objective:To explore the potential mechanism of sorafenib resistance associated long non-coding RNA (lncRNA-SRLR) promoted invasion and metastasis in U2OS osteosarcoma cells.Methods:We transfected U2OS cells with negative control lentivirus (LV-NC) or lncRNA-SRLR overexpressed lentivirus (LV-over/SRLR) particles. LV-NC and LV-over/SRLR stable transfected cells (U20S/NC and U20S/SRLR) were selected by primary cell culture medium containing puromycin. The mRNA expressions of lncRNA-SRLR and procollagen-lysine, procollagen-lysine 2-oxoglutarate 5-dioxygenase 2 (PLOD2) were detected by quantitative real-time polymerase chain reaction (qRT-PCR). The effect of lncRNA-SRLR on the invasion of U2OS cells were determined by wound-healing assay and Transwell migration assay. The effect of SRLR on the interleukin-6 (IL-6) secretion of U2OS cells was evaluated by enzyme-linked immunosorbent assay (ELISA) analysis. The subcellular distribution of SRLR in U2OS cells was detected by fluorescence in situ hybridization (FISH) analysis.The expression of PLOD2 in cells was detected by immunofluorescence (IF). The expressions of PLOD2 and focal adhesion kinase (FAK)/signal transducer and activator of transcription 3 (STAT3) signal pathway related proteins in U2OS/NC and U2OS/SRLR cells were detected by western blotting.Results:qRT-PCR assay showed that mRNA expressions of lncRNA-SRLR and PLOD2 in U2OS/SRLR cells were (3 964.97±0.05) and (2.77±0.11), respectively, significantly higher than those in U2OS/NC cells ( P<0.001 or P<0.01). The results of wound-healing and Transwell migration assay showed that over-expression of SRLR markedly promoted the invasion ability of U2OS cells ( P<0.05). The result of ELISA analysis showed that the IL-6 secretions in U2OS/NC or U2OS/SRLR cells were (125.38±11.22) pg/ml or (119.97±13.43) pg/ml, without statistical significance ( P>0.05). The subcellular distribution assay revealed that lncRNA-SRLR is predominately located in the nucleus. The result of IF showed that compared with U2OS/NC cells, the expression of PLOD2 was up-regulated in U2OS/SRLR cells. The result of western blotting showed that over-expression of SRLR significantly increased the expression levels of PLOD2, phosphorylation (p)-FAK and p-STAT3 in U2OS cells ( P<0.01). Conclusion:lncRNA-SRLR promotes invasion and metastasis of osteosarcoma by activating PLOD2-FAK/STAT3 signal axis.
10.Mechanism of lncRNA-SRLR induced invasion and metastasis in U2OS osteosarcoma cells
Fei CAO ; Xiaohong KANG ; Dafeng WANG ; Long MA ; Xiangjun CAO ; Ying WANG ; Yuanyuan GAO ; Zhanhui MIAO ; Haibin DENG ; Yabin GONG
Chinese Journal of Oncology 2020;42(12):1007-1013
Objective:To explore the potential mechanism of sorafenib resistance associated long non-coding RNA (lncRNA-SRLR) promoted invasion and metastasis in U2OS osteosarcoma cells.Methods:We transfected U2OS cells with negative control lentivirus (LV-NC) or lncRNA-SRLR overexpressed lentivirus (LV-over/SRLR) particles. LV-NC and LV-over/SRLR stable transfected cells (U20S/NC and U20S/SRLR) were selected by primary cell culture medium containing puromycin. The mRNA expressions of lncRNA-SRLR and procollagen-lysine, procollagen-lysine 2-oxoglutarate 5-dioxygenase 2 (PLOD2) were detected by quantitative real-time polymerase chain reaction (qRT-PCR). The effect of lncRNA-SRLR on the invasion of U2OS cells were determined by wound-healing assay and Transwell migration assay. The effect of SRLR on the interleukin-6 (IL-6) secretion of U2OS cells was evaluated by enzyme-linked immunosorbent assay (ELISA) analysis. The subcellular distribution of SRLR in U2OS cells was detected by fluorescence in situ hybridization (FISH) analysis.The expression of PLOD2 in cells was detected by immunofluorescence (IF). The expressions of PLOD2 and focal adhesion kinase (FAK)/signal transducer and activator of transcription 3 (STAT3) signal pathway related proteins in U2OS/NC and U2OS/SRLR cells were detected by western blotting.Results:qRT-PCR assay showed that mRNA expressions of lncRNA-SRLR and PLOD2 in U2OS/SRLR cells were (3 964.97±0.05) and (2.77±0.11), respectively, significantly higher than those in U2OS/NC cells ( P<0.001 or P<0.01). The results of wound-healing and Transwell migration assay showed that over-expression of SRLR markedly promoted the invasion ability of U2OS cells ( P<0.05). The result of ELISA analysis showed that the IL-6 secretions in U2OS/NC or U2OS/SRLR cells were (125.38±11.22) pg/ml or (119.97±13.43) pg/ml, without statistical significance ( P>0.05). The subcellular distribution assay revealed that lncRNA-SRLR is predominately located in the nucleus. The result of IF showed that compared with U2OS/NC cells, the expression of PLOD2 was up-regulated in U2OS/SRLR cells. The result of western blotting showed that over-expression of SRLR significantly increased the expression levels of PLOD2, phosphorylation (p)-FAK and p-STAT3 in U2OS cells ( P<0.01). Conclusion:lncRNA-SRLR promotes invasion and metastasis of osteosarcoma by activating PLOD2-FAK/STAT3 signal axis.

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