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Objective:To investigate the regulation of transcription factor CCCTC binding factor (CTCF) on the expression of B-cell lymphoma 2 ( Bcl-2) gene in pterygium and its molecular mechanism. Methods:Pterygium tissue samples from 22 primary pterygium patients who underwent pterygium excision combined with autologous limbal stem cell transplantation in The First Hospital of Changsha from June 2017 to February 2019 were collected during the operation as pterygium group.Normal conjunctival tissue from 20 patients with ocular trauma due to conjunctiva rupture, eyeball rupture or eyeball perforation in the same period were collected during the repair of ocular trauma as control group.Real-time PCR and Western blot were used to detect the expression levels of CTCF and Bcl-2 in the two groups.The DNA methylation level of the Bcl-2 promoter in the samples of the two groups was detected by bisulfite sequencing PCR (BSP). Pterygium fibroblasts were isolated and cultured.Fibroblasts were identified by immunohistochemistry using vimentin antibody.The cultured pterygium fibroblasts were divided into a CTCF interference group transfected with CTCF interference plasmid, and a control group transfected with control plasmid.The expression levels of CTCF and Bcl-2 in pterygium fibroblasts in CTCF interference and control groups were detected by real-time PCR and Western blot.The cell vitality was detected with cell counting kit-8 at 12, 24, and 48 hours after transfection.The DNA methylation level of the Bcl-2 promoter in the cells of the CTCF interference and control groups after transfection was determined by BSP.Differences of the indexes among groups were analyzed.Correlation between Bcl-2 mRNA and Bcl-2 gene promoter methylation level of CTCF protein in pterygium tissue was analyzed by Pearson linear correlation analysis.This study protocol was approved by the Ethics Committee of The First Hospital of Changsha (No.KL-2017021). Written informed consent was obtained from the patients from whom the specimens were collected.Results:The relative expression levels of CTCF mRNA and protein in pterygium group were 7.23±3.34 and 0.92±0.21, respectively, which were significantly higher than 1.10±0.44 and 0.28±0.07 in normal conjunctiva group ( t=-8.136, -13.025; both at P<0.01). The relative expression levels of Bcl-2 mRNA and protein in pterygium group were 10.27±4.64 and 0.95±0.27, which were higher than 1.10±0.41 and 0.32±0.14 in normal conjunctiva group, showing statistically significant differences ( t=-8.789, -10.782; both at P<0.01). The CTCF protein expression was significantly positively correlated with the Bcl-2 mRNA expression in pterygium group ( r=0.746, P<0.01). The DNA methylation level of the Bcl-2 promoter in pterygium group was 0.65±0.09, which was lower than 0.83±0.06 in normal conjunctiva group, with a statistically significant difference ( t=7.408, P<0.01). The DNA methylation level was significantly negatively correlated with the Bcl-2 mRNA expression in pterygium group ( r=-0.635, P<0.01). After the interference of CTCF expression in pterygium fibroblasts, the relative expression levels of CTCF and Bcl-2 mRNA in CTCF interference group were 0.37±0.03 and 0.53±0.06, which were significantly lower than 1.02±0.06 and 0.99±0.07 in control group ( t=20.035, 9.029; both at P<0.01). The relative expression levels of CTCF and Bcl-2 proteins in CTCF interference group were 0.23±0.06 and 0.56±0.07, which were lower than 0.52±0.05 and 0.92±0.12 in control group, showing statistically significant differences ( t=6.914, 4.719; both at P<0.01). The cell viability of pterygium fibroblasts in CTCF interference group was 0.10±0.01, 0.17±0.01, 0.38±0.04 at 12, 24, and 48 hours after interference, respectively, which were lower than 0.12±0.01, 0.29±0.01 and 0.85±0.06 in control group, and the differences were statistically significant ( t=3.718, 18.350, 15.621; all at P<0.01). The DNA methylation level of Bcl-2 promoter in CTCF interference group was 0.75±0.04, which was significantly higher than 0.61±0.03 in control group ( t=-4.472, P<0.05). Conclusions:CTCF is excessively expressed in pterygium, which may mediate the overexpression of Bcl-2 through down-regulating DNA methylation level.
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Objective:To observe the clinical curative e ect of treating diarrhea-predominant irritable bowel syndrome(D-IBS) with Lichang Tang(Intestine-Regulating Decoction) and sertraline hydrochloride.Methods:72 patients with(D-IBS) was randomly divided into 2 groups:32 cases in the treatment group was given self-prescribed Lichang Tang(modi ed Tongxie Yaofang [Main recipe for painful diarrhea]+Jinlingzi San [Toosendan Powder]) and sertraline hydrochloride(50 mg,p.o.once in the early morning);35 cases in the control group were given Lichang Tang alone for oral administration.During administering these drugs,other drugs were discontinued in the 2 groups.Psychological counseling and dietary modi cation were used in all patients.The course of treatment lasted for 4 weeks.Results:The total e ective rates in the treatment group and the control group were 86.49% and 65.71% respectively,and there was signi cant di erence(P
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Objective To evaluate the mid-long term effects of laparoscopic treatment of ovarian endometriosis cysts.Methods Clinical data of 209 cases of ovarian endometriosis cysts treated by laparoscopic operation from January 2002 to December 2005 were retrospectively analyzed.All the patients were followed for at least 12 months(12-54 months).There were 115 cases of unilateral ovarian cyst enucleation,62 cases of bilateral enucleation,11 cases of unilateral adnexectomy,and 21 cases of unilateral adnexectomy combined with contralateral cyst enucleation.Oral gestrinone was administrated in patients without childbearing intentions,while in patients demanding a child,no drug was given and an early pregnancy was encouraged.Results Conversions to open surgery were required in 7 cases.No complications occurred.Ovarian endometriosis cysts recurred in 17 cases(8.1%),including 7 cases of stage Ⅲendometriosis and 10 cases of stage Ⅳ endometriosis.Of the 35 cases of infertility,intrauterine pregnancy was obtained in 16 cases,with a pregnancy rate of 45.7%.The pregnancy occurred at 4-29 months after operation(within 2 years in 10 cases).Of the 157 cases of dysmenorrhea,a complete relief was achieved in 95 cases(60.5%)and an improvement was obtained in 62 cases(39.5%).Conclusions Laparoscopic treatment is suitable for all stages of ovarian endometriosis cysts.Postoperative drug administration and long-term follow-up are necessary to lower the recurrence rate.
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Objective To evaluate the mid-long term effects of hysteroscopic treatment of abnormal uterine bleeding(AUB).Methods A total of 432 patients admitted from January 2002 to November 2005 with AUB were treated with hysteroscopy.All the patients were followed for at least 12 months.Among the 432 patients,391 were diagnosed as having benign lesions,including endometrial polyp in 96 patients(22.2%),submucous myoma in 38 patients(8.8%),and endometrial hyperplasia in 257 patients(59.5%),receiving transcervical myoma resection or transcervical endometrium resection.The other 41 patients were diagnosed as having stage Ⅰ endometrial carcinoma,and a total hysterectomy and bilateral adnexectomy with selective pelvic lymphadenectomy was performed.Results All the operations were successfully completed,without uterine perforation or other complications.During a follow-up of 12～57 months(mean,39 months) in the 391 patients with benign lesions,amenorrhea was found in 63 patients(16.1%,63/391),hypomenorrhea( 1.5 cm in 8 patients,and a second hysteroscopic endometrial resection was given at 5～19 months after first operation.All the 8 patients were further followed for 18～36 months,with findings of amenorrhea in 5 of them and hypomenorrhea in 3.All the 41 patients with stage Ⅰ endometrial carcinoma survived without recurrence during a follow-up period for 26～41 months(mean,38 months).Conclusions Hysteroscopic treatment can be the first choice for treating non-malignant AUB.Pre-operational pathological examination and ultrasonography can enhance the safety and efficacy of the operation.