1.GDNF regulates the proliferation of glioma cells through AKT/β-catenin signaling pathway
Zhouru LI ; Daohui TENG ; Guokai DONG ; Wenjiang YIN ; Hongxing CAI
Chongqing Medicine 2015;(22):3034-3036
Objective To study the mechanism that glial cell line-derived neurotrophic factor (GDNF)promotes human glio-ma cells proliferation.Methods We divided glioma samples into two groups,including low-grade glioma group and high-grade glio-ma group,while cerebral contusion patients were treated as the control group,12 cases in each group.C6 glioma cell lines were di-vided into three groups,such as GDNF group,BSA(bovine serum albumin)group and control group.CCK-8 (cell counting kit-8) was used to detect the cell proliferation,while Western blot was used to detect the expression of AKT,p-AKT,β-catenin and p-β-catenin in each group.Results Comparing with the control group,the expression levels of AKT,p-AKT,β-catenin and p-β-catenin in glioma group had a significantly increased (P <0.05).Meanwhile,the high-grade gliomas group also had a significant increase in those more than low-grade gliomas group (P <0.05).CCK-8 test showed that the cell proliferation in GDNF group was significant-ly higher than the control group (P <0.05),and the expression levels of p-AKT,β-catenin and p-β-catenin proteins all had a signifi-cant increase (P <0.05).However,the expression level of AKT had no obvious difference.Conclusion GDNF might promote the proliferation of glioma cells by up-regulating the expression of p-AKT,β-catenin and p-β-catenin.
2.Expression of caspase-3 and HAX-1 after Cerebral Contusion in Rat
Zhouru LI ; Daohui TENG ; Guokai DONG ; Wenjiang YIN ; Hongxing CAI
Journal of Forensic Medicine 2015;(1):7-10,14
Objective To observe the expression pattern of caspase-3 and HCLS1-associated protein X-1 (HAX-1) at different time after cerebral contusion in rat, and explore the new method for estimating the injury interval. Methods The cerebral contusion model was established using adult SD male rats. Then the rats were randomly allocated into 8 groups: 2 h, 6 h, 12 h, 1 d, 3 d, and 7 d after cerebral con-tusion, sham-operation and normal control. Expression of caspase-3 and HAX-1 protein after cerebral contusion in rat was detected by Western blotting. Laser scanning confocal microscope was used to ob-serve the number of HAX-1 positive cells and TUNEL-stained cells after cerebral contusion. Results The expression of caspase-3 increased parallelly with the time after cerebral contusion and reached the peak value on 3 d. The expression of caspase-3 decreased gradually and still maintained a high level expression on 7 d (P<0.05). The expression of HAX-1 positive cell went up after injury, and reached the peak value at 6 h (P<0.05), then turned down gradually after 12 h and went out of detection after 3 d. The number of TUNEL-stained cells increased obviously at 2 h and reached the peak value on 3 d. The number of TUNEL-stained apoptotic cells decreased gradually and still maintained a high level expres-sion on 7 d (P<0.05). Conclusion The expression of caspase-3 and HAX-1 after cerebral contusion has time sequential regularity, which may provide new evidence for forensic diagnosis of cerebral contusion interval.
3.Increased Egr-1 binding to promoter induced by histone hyperacetylation promotes gdnf gene transcription.
Zhouru LI ; Jie LIU ; Yu LEI ; Haibo NI ; Hongxing CAI ; Baole ZHANG
Journal of Southern Medical University 2015;35(5):697-701
OBJECTIVETo investigate the mechanism of high transcription of the glial cell-line derived neurotrophic factor (gdnf) gene induced by hyperacetylation of histone H3 lysine 9 (H3K9) at its promoter region II in rat C6 glioma cells.
METHODSThe acetylation level of H3K9 at Egr-1 binding site in gdnf gene promoter region II and the binding capacity of Egr-1 to its binding site in gdnf promoter were examined by ChIP-PCR in C6 astroglioma cells and normal rat astrocytes, and its changes were investigated in C6 astroglioma cells after treatment with histone acetyltransferase inhibitor curcumin or deacetylase inhibitor trichostatin A.
RESULTSCompared normal astrocytes, C6 astroglioma cells showed significantly increased acetylation level of H3K9 at Egr-1 binding site in gdnf gene promoter region II and Egr-1 binding capacity (P<0.01). Curcumin treatment significantly reduced H3K9 acetylation level at Egr-1 binding site and decreased both the binding of Egr-1 to promoter region II and gdnf mRNA levels in C6 astroglioma cells (P<0.05). Conversely, increased H3K9 acetylation at the Egr-1 binding site induced by trichostatin A significantly increased the binding of Egr-1 to promoter region II and gdnf mRNA expression levels (P<0.05).
CONCLUSIONH3K9 hyperacetylation induces increased Egr-1 binding to gdnf gene promoter II, which might be the reason for the high transcription level of gdnf gene in rat C6 glioma cells.
Acetylation ; Animals ; Astrocytes ; metabolism ; Binding Sites ; Cell Line, Tumor ; Early Growth Response Protein 1 ; metabolism ; Glial Cell Line-Derived Neurotrophic Factor ; genetics ; Glioma ; metabolism ; Histones ; chemistry ; Promoter Regions, Genetic ; Protein Processing, Post-Translational ; RNA, Messenger ; Rats ; Transcription, Genetic
4.Research practice of using two heating blankets in combination to affect intraoperative temperature and postoperative resuscitation in patients undergoing endoscopic three-incision radical resection of esophageal cancer
Yanfang BAI ; Mingtian LI ; Zhouru ZHAN ; Huiying QIN
Chinese Journal of Practical Nursing 2020;36(33):2600-2606
Objective:To compare the effects of three heating methods on intraoperative temperature and postoperative resuscitation in patients undergoing three-incision radical resection of esophageal cancer under full endoscope.Methods:A total of 145 patients undergoing endoscopic three-incision radical resection of esophageal cancer were randomized to receive forced-air warming respectively on the under-body blanket(UB group, n=48), over-body blanket (OB group, n=48), underbody blanket in combination with over-body blanket(Combined group, n=49). The nasopharyngeal temperature were recorded at the time of entering the operating room, at the beginning of the intubation, at the beginning of the operation and every 30min after the operation, at the end of the operation. The incidence of hypothermia, hypothermia related adverse reactions and postoperative resuscitation indicators of the three groups were compared. Results:During the operation, the patients' temperature fluctuation showed two processes of decrease and two processes of increase. The temperature of Combined group showed a lower fall and a faster rise. The average temperature of the Combined group were higher than the other two group at each time point from the beginning of the intubation, the incidence of hypothermia was 4.08% (2/49), which was lower than the OB group [22.92% (11/48)] and the UB group [18.75% (9/48)], The difference was statistically significant ( χ2 value was 7.397, P=0.025). The temperature of the UB group was (36.52±0.18),(36.31±0.35)℃ at the beginning of intubation and at the end of the operation, respectively, which were higher than that of the OB group (36.44±0.15),(36.13±0.32)℃ ( t value was 2.393, 3.723, P < 0.05). The temperature comfort score of the combined group was 7.81±0.52, higher than that of the other two groups ( F value was 19.962, P<0.01), and the scores of chills, agitation and chills in the recovery period were lower than that of the other two groups ( F value was 8.186, 6.705, 4.051, all P < 0.05). The extubation time and the waking time of the combined group was (15.90±2.97)min, (31.47±4.42)min, respectively, which were both lower than those of the other two groups ( F value was 69.094, 114.549, P < 0.01). Conclusions:In endoscopic three-incision radical resection of esophageal cancer, combined heating makes the patients' temperature rise rapidly to offset the loss of heat, which is conducive to maintain a stable temperature. At the same time, the incidence of intraoperative hypothermia, postoperative chills and agitation is reduced, the temperature comfort of patients is improved, the extubation time and waking time are shortened, which is conducive to promote postoperative recovery of patients, thus reduce postoperative hypothermia related complications.
5.Increased Egr-1 binding to promoter induced by histone hyperacetylation promotes gdnf gene transcription
Zhouru LI ; Jie LIU ; Yu LEI ; Haibo NI ; Hongxing CAI ; Baole ZHANG
Journal of Southern Medical University 2015;(5):697-701
Objective To investigate the mechanism of high transcription of the glial cell-line derived neurotrophic factor (gdnf) gene induced by hyperacetylation of histone H3 lysine 9 (H3K9) at its promoter region II in rat C6 glioma cells. Methods The acetylation level of H3K9 at Egr-1 binding site in gdnf gene promoter region II and the binding capacity of Egr-1 to its binding site in gdnf promoter were examined by ChIP-PCR in C6 astroglioma cells and normal rat astrocytes, and its changes were investigated in C6 astroglioma cells after treatment with histone acetyltransferase inhibitor curcumin or deacetylase inhibitor trichostatin A. Results Compared normal astrocytes, C6 astroglioma cells showed significantly increased acetylation level of H3K9 at Egr-1 binding site in gdnf gene promoter region II and Egr-1 binding capacity (P<0.01). Curcumin treatment significantly reduced H3K9 acetylation level at Egr-1 binding site and decreased both the binding of Egr-1 to promoter region II and gdnf mRNA levels in C6 astroglioma cells (P<0.05). Conversely, increased H3K9 acetylation at the Egr-1 binding site induced by trichostatin A significantly increased the binding of Egr-1 to promoter region II and gdnf mRNA expression levels (P<0.05). Conclusion H3K9 hyperacetylation induces increased Egr-1 binding to gdnf gene promoter II, which might be the reason for the high transcription level of gdnf gene in rat C6 glioma cells.
6.Increased Egr-1 binding to promoter induced by histone hyperacetylation promotes gdnf gene transcription
Zhouru LI ; Jie LIU ; Yu LEI ; Haibo NI ; Hongxing CAI ; Baole ZHANG
Journal of Southern Medical University 2015;(5):697-701
Objective To investigate the mechanism of high transcription of the glial cell-line derived neurotrophic factor (gdnf) gene induced by hyperacetylation of histone H3 lysine 9 (H3K9) at its promoter region II in rat C6 glioma cells. Methods The acetylation level of H3K9 at Egr-1 binding site in gdnf gene promoter region II and the binding capacity of Egr-1 to its binding site in gdnf promoter were examined by ChIP-PCR in C6 astroglioma cells and normal rat astrocytes, and its changes were investigated in C6 astroglioma cells after treatment with histone acetyltransferase inhibitor curcumin or deacetylase inhibitor trichostatin A. Results Compared normal astrocytes, C6 astroglioma cells showed significantly increased acetylation level of H3K9 at Egr-1 binding site in gdnf gene promoter region II and Egr-1 binding capacity (P<0.01). Curcumin treatment significantly reduced H3K9 acetylation level at Egr-1 binding site and decreased both the binding of Egr-1 to promoter region II and gdnf mRNA levels in C6 astroglioma cells (P<0.05). Conversely, increased H3K9 acetylation at the Egr-1 binding site induced by trichostatin A significantly increased the binding of Egr-1 to promoter region II and gdnf mRNA expression levels (P<0.05). Conclusion H3K9 hyperacetylation induces increased Egr-1 binding to gdnf gene promoter II, which might be the reason for the high transcription level of gdnf gene in rat C6 glioma cells.
7.A comparative study between retro-auricular single-site endoscopic thyroidectomy and transoral endoscopic thyroidectomy vestibular approach: a single-center retrospective analysis
Fan DONG ; Yong AO ; Mingtian LI ; Zhouru ZHAN ; Yongqin LIN ; Qingjian TAN ; Hao LI ; Ankui YANG ; Dian OUYANG
Chinese Journal of Surgery 2021;59(11):891-896
Objectives:To compare the efficiacy of retro-auricular single-site endoscopic thyroidectomy (RASSET) and that of transoral endoscopic thyroidectomy vestibular approach (TOETVA).Methods:In Department of Head and Neck Surgery, Sun Yat-sen University Cancer Center, 10 patients underwent RASSET from June 2021 to August 2021, and 21 patients underwent TOETVA from January 2016 to August 2021. All the 21 patients′ clinical data was analyzed retrospectively. There were 2 males and 8 females in the RASSET group, aging (48.2±13.9) years (range: 28 to 67 years). There were 5 males and 16 females in the TOETVA group, aging (31.3±8.2) years (range: 21 to 49 years). All patients underwent thyroid lobectomy. A 3 cm in length incision was cut on single auricula posterior sulci to creat the approach in the RASSET group. Then a Trocar made with of a glove was inserted. Retaining the omohyoid, the sternocleidomastoid muscle and anterior cervical muscle were pulled apart, exposing a single lobe of the thyroid gland and lymphatic tissue of zone Ⅵ, for en-bloc resection. The clinical data of the two groups were collected and analyzed by t test, Mann-Whitney U test, Fisher exact test or χ 2 test. Clinical data and postoperative efficacy indexes such as operation time, postoperative C reactive protein level, and postoperative complications were recorded. Results:Compared with the TOETVA group, the operation time was longer in the RASSET group ((256.8±77.0) minutes vs. (201.2±54.9) minutes, t=2.31, P=0.028), and increase of postoperative C reaction protein (24 hours postoperative vs. preoperative) was lower in the RASSET group (8.58(13.24) mg/L vs. 46.24(48.88) mg/L, Z=-4.311, P<0.01). But there was no significant difference between the RASSET group and TOETVA group in the number of lymph nodes dissection (2(5) vs. 2(3), Z=-0.326, P=0.759). Besides, there were no complications in the RASSET group. Conclusion:Retro-auricular single-site endoscopic thyroid loectomy is easy to achieve the en-bloc resection of tumors with a well-concealed scar and less traumatic dissection.
8.A comparative study between retro-auricular single-site endoscopic thyroidectomy and transoral endoscopic thyroidectomy vestibular approach: a single-center retrospective analysis
Fan DONG ; Yong AO ; Mingtian LI ; Zhouru ZHAN ; Yongqin LIN ; Qingjian TAN ; Hao LI ; Ankui YANG ; Dian OUYANG
Chinese Journal of Surgery 2021;59(11):891-896
Objectives:To compare the efficiacy of retro-auricular single-site endoscopic thyroidectomy (RASSET) and that of transoral endoscopic thyroidectomy vestibular approach (TOETVA).Methods:In Department of Head and Neck Surgery, Sun Yat-sen University Cancer Center, 10 patients underwent RASSET from June 2021 to August 2021, and 21 patients underwent TOETVA from January 2016 to August 2021. All the 21 patients′ clinical data was analyzed retrospectively. There were 2 males and 8 females in the RASSET group, aging (48.2±13.9) years (range: 28 to 67 years). There were 5 males and 16 females in the TOETVA group, aging (31.3±8.2) years (range: 21 to 49 years). All patients underwent thyroid lobectomy. A 3 cm in length incision was cut on single auricula posterior sulci to creat the approach in the RASSET group. Then a Trocar made with of a glove was inserted. Retaining the omohyoid, the sternocleidomastoid muscle and anterior cervical muscle were pulled apart, exposing a single lobe of the thyroid gland and lymphatic tissue of zone Ⅵ, for en-bloc resection. The clinical data of the two groups were collected and analyzed by t test, Mann-Whitney U test, Fisher exact test or χ 2 test. Clinical data and postoperative efficacy indexes such as operation time, postoperative C reactive protein level, and postoperative complications were recorded. Results:Compared with the TOETVA group, the operation time was longer in the RASSET group ((256.8±77.0) minutes vs. (201.2±54.9) minutes, t=2.31, P=0.028), and increase of postoperative C reaction protein (24 hours postoperative vs. preoperative) was lower in the RASSET group (8.58(13.24) mg/L vs. 46.24(48.88) mg/L, Z=-4.311, P<0.01). But there was no significant difference between the RASSET group and TOETVA group in the number of lymph nodes dissection (2(5) vs. 2(3), Z=-0.326, P=0.759). Besides, there were no complications in the RASSET group. Conclusion:Retro-auricular single-site endoscopic thyroid loectomy is easy to achieve the en-bloc resection of tumors with a well-concealed scar and less traumatic dissection.