1.Research on the expression of Survivin gene suppressed by pre-irradiation and intratumor injection siRNA in lung cancer of mouse
Lihua WANG ; Chao YAN ; Rongrong LI ; Wei LU
China Oncology 2015;(5):339-344
Background and purpose: The effects of tumor after radiotherapy and chemotherapy with tumor cell apoptosis have positive correlation. Survivin is an apoptosis suppressor gene, which is overexpressed in lung cancer. The reduction of its expression can increase lung cancer cell apoptosis. RNA interference can speciifcally and effectively blocked the expression of this gene. After accepting a dose of radiation, tumor tissues can raise the rate of gene transduction. The purpose of this study was to investigate the effects of the expression of Survivin gene suppressed by intratumor injection siRNA and radiation in lung cancer. Methods:Mouse subcutaneous transplanted with tumors were randomly divided into four groups:untreated group (group A), group with intratumor injection siRNA (group B), group with radiation (group C), group 4 Gy pre-irradiation combination with intratumor injection siRNA (group D). Two days later, mouse with different treatments were executed by cervical dislocation and stripped the subcutaneous tumors. mRNA and protein levels of Survivin gene were detected by quantitative real-time PCR (qRT-PCR) and Western blot; Cell cycle and cell apoptosis were assayed by lfow cytometry (FCM). Results:The expression of Survivin gene in mRNA and protein levels in group B, C, and D were signiifcantly reduced compared with group A. The differences of Survivin expression in mRNA level between group A and group D were statistically signiifcant. Compared with the other 3 groups, the expression of Survivin gene in group D was signiifcantly reduced, the differences were statistically signiifcant (P=0.036);The proportion of cells in S-phase of group D was (2.70±0.34)%, compared with group B [(8.93±0.75)%] and group C [(6.71±0.51)%], that was significantly reduced. The differences of the proportion of S-phase cells in group A and group D were statistically signiifcant (P=0.034);The rate of cell apoptosis in group D was (25.67 ± 0.65)%, which was signiifcantly increased compared with the rate of cell apoptosis in the other 3 groups, the differences were statistically signiifcant (F=78.82, P<0.05). Conclusion:Pre-irradiation can enhance the transduction rate of siRNA, reduce the expression of Survivin gene in lung cancer, promote cell apoptosis, and increase the sensitivity of the radiotherapy.
2.Measurement of the angular velocity and perpendicular ground reaction force of the ankle joint in parachute landing simulation
Chao ZHENG ; Ji WU ; Rongrong HUANG ; Songchao CUI ; Yanwu WEN ; Yi LI ; Di WU
Chinese Journal of Orthopaedics 2014;34(6):688-693
Objective To measure the angular velocity and perpendicular ground reaction force of the ankle joint under different heights with half-squat jumping in parachute training simulation,providing a reliable experiment basis for the preventing of ankle injury.Methods A total of 18 volunteers participated in this study.The experimental group included 9 male with experience of parachute landing,while the other 9 male without experience of parachute landing were assigned to the control group.Each subject was instructed to jump off a platform with a height of 30 cm and 60 cm and land on a hard surface in a half-squat posture.The dynamic landing process was recorded with a high speed camera and the biomechanical data was collected and analyzed,including perpendicular ground reaction force,angular displacement,velocity and acting time.Results From 30 cm's height,the ankle angular displacement of the control group was significantly larger than the experimental group (25.73°± 8.13° vs 20.05°± 12.27°,P < 0.05).The perpendicular ground reaction force of the control group was significantly smaller than the experimental group (3 372.4±748.6 N vs 5 181.5±1 726.2 N,P < 0.05).The acting time of the control group was significantly longer than the ex perimental group (0.049±0.015 s vs 0.012±0.004 s,P < 0.05).The buffer time of the control group was significantly shorter than the experimental group (1.397±0.746 s vs 1.737±0.451 s,P < 0.05).From 60 cm's height,the ankle angular velocity of the control group was significantly higher than the experimental group (25.45± 15.01 °/s vs 16.51 ±4.18 °/s,P < 0.05).The perpendicular ground reaction force of the control group was significantly smaller than the experimental group (4 616.0±1 124.7 N vs 7 119.5±2 307.4 N,P < 0.05).The acting time of the control group was significantly longer than the experimental group (0.048±0.013 s vs 0.015±0.006 s,P < 0.05).The buffer time of the control group was significantly shorter than the experimental group (0.922±0.347 s vs 1.617±0.547 s,P < 0.05).Conclusion Jumping from different heights,the experinental group was larger in perpendicular ground reaction force but smaller in the angular velocity and displacement than the control group.There was a shorter acting time and a longer buffer time in the experimental group than the control group.
3.Interleukin-1 receptor type 1 signaling induces excessive inflammatory responses in H1N1 influenza ;virus infection
Rongrong REN ; Xiaonan REN ; Boyin QIN ; Mengjiao YUAN ; Hua YANG ; Chao WANG ; Shun LI ; Xiaohui ZHOU
Chinese Journal of Microbiology and Immunology 2016;36(12):887-893
Objective To investigate the role of interleukin-1 receptor type 1 (IL-1R1) signaling in H1N1 influenza virus infection. Methods IL-1R1 knockout ( IL-1R1-/-) mice and wild type ( WT) mice were infected intranasally with 2×104 TCID50(50% tissue culture infective dose) of influenza virus H1N1 PR8. Changes in clinical signs, survivals and bodyweights of those mice were monitored daily for 14 consecutive days. Three mice from each group were sacrificed at 3, 7 and 14 days post infection (d. p. i), from which whole lungs were harvested. A part of the lobes was fixed in 4% paraformaldehyde for histopatho-logical assessment and the rest were split and stored at-80 centigrade for further analysis. Real-time quanti-tative PCR and cytometric bead array ( CBA) were performed to detect viral loads in lungs and inflammatory cytokines in supernatants of lung homogenates. Results The mice in both groups showed severe symptoms after the infection of PR8. The maximum bodyweight loss of IL-1R1-/- mice [(24. 22±0. 80) % at 8 d. p. i] was lower than that of WT mice [(28. 03±1. 51)% at 9 d. p. i] (P<0. 05). The IL-1R1-/- mice with PR8 infection showed a higher survival rate (90%) as compared with that of the control group (40%) (P<0. 05). No statistical differences in virus loads were observed between the two groups at 3, 7 and 14 d. p. i. The lung weight to body weight ratio of IL-1R1-/-mice [(1. 42±0. 03) %] was lower than that of WT mice [(1. 79±0. 08) %] at 3 d. p. i (P<0. 05). Pathological changes in IL-1R1-/- mice were less severe than those in WT mice. CBA detection assay revealed that the proinflammatory cytokines in lungs of IL-1R1-/-mice were less than those in WT mice. Conclusion IL-1R1 signaling plays a pathogenic role in mice infec-ted with 2×104 TCID50 of influenza virus PR8 by promoting inflammatory responses.
4.Establishment of comprehensive evaluation indexing system and weight values on performing HIV/AIDS prevention
Qin XIAO ; Pinyi CHEN ; Guohui WU ; Rongrong LU ; Chao ZHOU ; Ling LIU ; Yanqi ZHANG ; Zhonghong YAN ; Dong YI
Chongqing Medicine 2013;(28):3408-3410
Objective To establish a comprehensive evaluation indexing system to appraise the implications of prevention and treatment of HIV/AIDS ,and to calculate the weight of each indicator .Methods Based on the idea of performance and input-out-put ,professional consultation ,and Delphi method was determined as the evaluation index system ,analytical hierarchy process (AHP) was used to calculate the weight value for each indicator .Results The evaluation indexing system had been established af-ter three rounds of professional consultation .It contained two 1st class indicators ,six 2nd class indicators and thirty-one in 3rd class indicators .The weight value of each indicator was calculated .Conclusion The evaluation indexing system that has been established and the weight value quantities are of completeness ,practicality ,operability and logic .They have important value for application in the future .
5.Artificial cervical disc replacement: range of motion of replacement segment and degeneration of adjacent segments
Xuanyu CHEN ; Ji WU ; Chao ZHENG ; Rongrong HUANG ; Yuming CUI ; Yong SHANG ; Henghua FAN ; Panfeng YU ; Xuhong ZHAO ; Dong CHU
Chinese Journal of Tissue Engineering Research 2015;19(17):2672-2676
BACKGROUND:In recent years,artificial cervical disc replacement surgery as a new method for the treatment of cervical disease has gradualy been accepted and understood,but relevant complications have gradualy attracted attention.OBJECTIVE:To investigate the clinical outcomes of artificial cervical disc replacement in the treatment of cervical disease and the range of motion of the replacement segment.METHODS: A total of 25 patients with artificial cervical disc replacement in the treatment of cervical spondylosis,who were treated in the Department of Orthopedics,Air Force General Hospital of Chinese PLA from August 2006 to April 2012,were enroled in this study,including 15 males and 10 females,aged 31-76 years,averagely 51.04 years.There were 6 cases of double segments and 19 cases of single segment.They were folowed up for 24 to 93 months.Clinical results were assessed using the Japanese Orthopaedic Association score,cervical dysfunction index and pain visual analog scale scores.Imaging was used to observe range of motion,cervical curvature,heterotopic ossification,and degeneration of adjacent segments.RESULTS AND CONCLUSION:Neurological function in al patients was improved to different degrees.One case suffered from mild heterotopic ossification,but no clinical symptoms were found.No significant difference in range of motion of surgical segment,and range of motion of upper and lower adjacent segments was detected between pre-replacement and final folow-up results (P>0.05).No significant difference in range of motion of C2-C7 was found between pre-replacement and final folow-up results (P>0.05).Japanese Orthopaedic Association score,cervical dysfunction index and pain visual analog scale scores were significantly improved during final folow-up compared with pre-replacement (P<0.05).These results indicated that artificial cervical disc replacement in the treatment of cervical disease can achieve better clinical efficacy,can keep the range of motion of replacement segment and avoid the accelerated degeneration of adjacent segments.
6.MicroRNA-708-3p as a potential therapeutic target via the ADAM17-GATA/STAT3 axis in idiopathic pulmonary fibrosis
Bo LIU ; Rongrong LI ; Jinjin ZHANG ; Chao MENG ; Jie ZHANG ; Xiaodong SONG ; Changjun LV
Experimental & Molecular Medicine 2018;50(3):e465-
MicroRNAs (miRNAs) are important diagnostic markers and therapeutic targets for many diseases. However, the miRNAs that control the pathogenesis of idiopathic pulmonary fibrosis (IPF) and act as potential therapeutic targets for the disease are rarely studied. In the present study, we analyzed the function and regulatory mechanism of microRNA-708-3p (miR-708-3p) and evaluated this marker’s potential as a therapeutic target in IPF. The clinical and biological relevance of fibrogenesis for miR-708-3p was assessed in vivo and in vitro, specifically in matching plasma and tissue samples from 78 patients with IPF. The data showed that the miR-708-3p levels decreased during fibrosis and inversely correlated with IPF. The experiments showed that the decreased miR-708 promoter activity and primer-miR-708(pri-miR-708) expression were the potential causes. By computational analysis, a dual luciferase reporter system, rescue experiments and a Cignal Finder 45-Pathway system with siADAM17 and a miR-708-3p mimic, we identified that miR-708-3p directly regulates its target gene, a disintegrin and metalloproteinase 17 (ADAM17), through a binding site in the 3′ untranslated region, which depends on the GATA/STAT3 signaling pathway. Finally, an miR-708-3p agomir was designed and used to test the therapeutic effects of the miR-708-3p in an animal model. Small-animal imaging technology and other experiments showed that the dynamic image distribution of the miR-708-3p agomir was mainly concentrated in the lungs and could block fibrogenesis. In conclusion, the miR-708-3p–ADAM17 axis aggravates IPF, and miR-708-3p can serve as a potential therapeutic target for IPF.
7.Effects and mechanism of insulin-like growth factor 1 on the secretion of transforming growth factor β2 and matrix metalloproteinase 2 in human reti-nal pigment epithelial cells
Rongrong CHAO ; Liu ZHENG ; Jing FAN ; Zhixiang DING
Recent Advances in Ophthalmology 2024;44(7):512-517
Objective To investigate the effects of insulin-like growth factor 1(IGF-1)on the expression of transfor-ming growth factor β2(TGF-β2)and matrix metalloproteinase 2(MMP-2)in human retinal pigment epithelial cells(ARPE-19)and related mechanisms.Methods ARPE-19 cells were cultured for 6 h,12 h,24 h and 48 h,respectively,with dif-ferent concentrations of IGF-1 and LY294002.The cell viability was detected using the cell counting kit-8 to determine the optimal action concentration and time of IGF-1 and LY294002.The cell migration activity was detected using the cell scratch assay.The concentration of TGF-β2 in cell culture supernatant was detected using the enzyme-linked immunosorbent assay(ELISA).ARPE-19 cells were divided into the control group,IGF-1 group(80 μg·L-1 IGF-1),IGF-1+LY294002 group(80 μg·L-1 IGF-1+30 mmol·L-1 LY294002),and LY294002 group(30 mmol·L-1 LY294002)and cultured with serum-free DMEM/F12 medium,while cells in the control group received no treatment.The mRNA and protein expression levels of TGF-β2,MMP-2,phosphoinositide 3-kinase(PI3K)and protein kinase B(AKT)in the cells were measured using the re-verse transcription-polymerase chain reaction(RT-PCR)and Western blot,respectively.Results Compared with the 0μg·L-1 IGF-1 group,the cell viability in the 80 μg·L-1 IGF-1 group changed the most significantly at 24 h(P<0.05);thus,the optimal concentration of IGF-1 was 80 μg·L-1 and the optimal culture time was 24 h.Compared with the 0 mmol·L-1 LY294002 group,the inhibition concentration in the 30 mmol·L-1 LY294002 group at 24 h was close to half;thus,the optimal concentration of LY294002 was 30 mmol·L-1 and the optimal culture time was 24 h.The cell scratch assay re-sults showed that the cell migration rate was significantly different among the 0 μg·L-1 IGF-1 group,40 μg·L-1 IGF-1 group,and 80 μg·L-1 IGF-1 group(all P<0.05).ELISA results showed that there was a statistically significant difference in the concentration of TGF-β2 in cell supernatant among the 0 μg·L-1 IGF-1 group,40 μg·L-1 IGF-1 group,and 80 μg·L-1 IGF-1 group(all P<0.05).RT-PCR and Western blot results showed that after 24 h culture with IGF-1 and LY294002,compared with the control group,the mRNA and protein expression levels of TGF-β2,MMP-2,PI3K and AKT in the IGF-1 group increased,while the mRNA and protein expression levels of TGF-β2,MMP-2,PI3K and AKT in the LY294002 group decreased(all P<0.05).Compared with the IGF-1 group,the mRNA and protein expression levels of TGF-[32,MMP-2,PI3K and AKT in the IGF-1+LY294002 group decreased(all P<0.05).Conclusion IGF-1 can promote the proliferation and migration of ARPE-19 cells.IGF-1 may up-regulate the expression of TGF-β2 and MMP-2 in ARPE-19 cells through the PI3K/AKT signaling pathway and participate in the occurrence and development of myopia.
8.Endovascular therapy combined with laparoscopic surgery for acute mesentery artery occlusion
Keqiang ZHAO ; Peng ZHANG ; Junlai ZHAO ; Tong ZHANG ; Zhanjiang CAO ; Yu YANG ; Chao JIANG ; Rongrong ZHU ; Weiwei WU
Chinese Journal of General Surgery 2024;39(3):192-196
Objective:To explore the efficacy and safety of endovascular intervention combined with preoperative laparoscopic exploration in the treatment of patients with acute mesenteric artery ischemia.Methods:This was a prospective cohort study (NCT04686981). The study enrolled 31 patients with acute mesenteric artery ischemia from Oct 1, 2020 to Oct 1, 2022. Among them, 26 patients (84%) were male, with a mean age of (67±13) years and a mean time to onset of (21±8) hours. All patients underwent laparoscopic exploration in the hybrid operating room. If the presence of intestinal necrosis or suspected necrosis was clearly determined, the patient would undergo open surgery (mesenteric artery embolization, intestinal resection and intestinal double stoma) as the treatment by gastrointestinal surgeon. If intestinal necrosis or suspected necrosis was not found by laparoscopy, the patient would undergo endovascular intervention by vascular surgeon. The primary observational endpoints of this study were the proportion of patients who were not dependent on total parenteral nutrition and all-cause mortality within 30 days after operation. The secondary observational endpoints were the rate of mesenteric vascular patency within 30 days and the proportion of interventions that were converted to open surgery.Results:Six patients underwent open surgery and 25 patients underwent endovascular intervention, including 13 cases of thrombus reduction alone, 3 cases of stent implantation during the same period after reduction, and 9 cases of stent implantation alone. Twenty-four patients (77%) were completely weaned from the TPN within 30 days after the procedure, and all-cause mortality was observed in 3 cases (9.7%). The patency rate of the mesenteric artery within 30 days after the procedure was 82.1%. The rate of conversion to open surgery after intervention was 16%.Conclusions:Endovascular intervention combined with preoperative laparoscopic exploration can clarify intestinal ischemia in acute mesenteric patients as early as possible, and individualized treatment strategies for each patient by multidisciplinary care team can potentially improve the prognosis of such patients.
9.Effect of intraoperative continuous infusion insulin on myocardial perfusion in patients after cardiac surgery under cardiopulmonary bypass
Zhuan ZHANG ; Jiajia YIN ; Ning LI ; Chao CHEN ; Kai ZHANG ; Rongrong MA ; Shiyu GUAN ; Jianyou ZHANG ; Qiang WANG ; Hu LI
The Journal of Clinical Anesthesiology 2024;40(5):497-502
Objective To investigate the effect of continuous intraoperative insulin infusion on my-ocardial blood perfusion after cardiac surgery under cardiopulmonary bypass(CPB).Methods Forty-eight patients,21 males and 27 females,aged 55-80 years,BMI 18-28 kg/m2,ASA physical status Ⅱ-Ⅳ,who underwent elective cardiac surgery with CPB were selected and randomly divided into two groups:the insulin group(group I,n = 25)and the control group(group C,n = 23).The same anesthesia protocol was implemented in both groups.After induction of anesthesia,group Ⅰ received intravenously infusion of in-sulin 30 mU·kg-1·h-1,glucose 0.12 g·kg-1·h-1,and potassium chloride 0.06 mmol·kg-1·h-1,and group C received saline 10 ml/h,all of which were infused until the end of surgery.The targeted blood glucose range for both groups was set at 6.1-11.1 mmol/L.Transesophageal echocardiography(TEE)was performed 10 minutes after induction of general anesthesia(T2)and before the end of surgery(T6)to ex-amine the coronary sinus(CS)flow spectrum and diameter,pulmonary venous flow spectrum,and calculate CS net antegrade flow velocity time integral(VTI).Femoral mean arterial pressure(MAP),central venous pressure(CVP),stroke volume(SV),cardiac index(CI)and peripheral vascular resistance index(SVRI)were recorded at T2,2 minutes before CPB(T3),the end of CPB(T5),and T6.The concentra-tions of blood glucose and lactate 5 minutes before anesthesia induction(T1),T3,30 minutes after CPB(T4),T5,T6,6 hours after surgery(T7),12 hours after surgery(T8),and 24 hours after surgery(T9)were recorded.The levels of high-sensitivity C-reactive protein(hs-CRP),high-sensitivity troponin I(hs-TnI),and creatine kinase isoenzyme(CK-MB)were recorded 1 day preoperatively,1 and 2 days post-operatively.Results Compared with group C,in group I,CS net antegrade flow VTI and blood flow per minute were significantly increased(P<0.05),and pulmonary venous peak atrial reversal wave velocity(ARp)was significantly reduced at T6(P<0.05),SV and CI were significantly increased and SVRI was significantly decreased at T5 and T6(P<0.05),lactate concentration was significantly decreased at T7 and T8(P<0.05),hs-CRP and CKMB were significantly decreased 1 and 2 days postoperatively(P<0.05),hs-TnI was significantly reduced 2 days postoperatively(P<0.05).Conclusion Continuous insulin admin-istration during cardiac surgery with CPB while maintaining blood glucose at 6.1-11.1 mmol/L can enhance myocardial blood perfusion,mitigate postoperative inflammatory response,and reduce myocardial injury.
10.Experience and early results of radiofrequency ablation therapy for varicose veins
Junlai ZHAO ; Weiwei WU ; Keqiang ZHAO ; Zhanjiang CAO ; Yu YANG ; Chao JIANG ; Rongrong ZHU
Chinese Journal of General Surgery 2019;34(7):598-600
Objective To evaluate the experience and early results of radiofrequency ablation (RFA) therapy for varicose veins.Methods In this study 380 extremities of 356 patients undergoing RFA therapy with RFA catheter for varicose veins from Nov 2015 to Sep 2017 in Beijing Tsinghua Changgung Hospital were retrospectively reviewed.Each patient was scheduled to follow up at 1,6 and 12 months respectively.Results Technical success were achieved in all cases.Obliteration rate was 99.0% in 1 month,97.9% in 6 months and 97.1% in 12 months after the RFA therapy.Conclusions RFA therapy for varicose veins is safe and effective and achieves good early result.Intraoperative ultrasound examination and endovascular technique is important for a successful RFA procedure.