1.Growth inhibition and radiosensitization of Celecoxib in nasopharyngeal carcinoma cell line CNE-2
Xinhua XU ; Fang YI ; Xiangyang FU ; Jingtao DU ; Xiaohong ZHANG ; Changju ZHANG ; Yanlin WANG
Chinese Journal of Radiation Oncology 2009;18(3):249-253
Objective To investigate the growth inhibition and radiosensitization of Celecoxib in hu-man nasopharyngeal carcinoma cell line CNE-2. Methods CNE-2 growth inhibition by Celecoxib was eval-uated by MTT method. Apoptosis-related changes in morphology were observed by transmission electron mi-croscopy (TEM). Cell cycle distribution and apoptosis rate were measured by flowcytometry (FCM). The ex-pression of COX-2 protein was observed by SP method after the treatment of Celecoxib. Cells were randomly planted into four groups: irradiation control(Ci), drug group(Cd), irradiation group(R), and Celecoxib plus irradiation group(D+R). Single irradiation of 2,4,6,8,and 10 Gy were administered for colonogenic assay. Cell cycle distribution and apoptosis rate were analyzed at 6 Gy irradiation. Results The growth of CNE-2 cell was inhibited by celecoxib in a dose-and time-dependent manner, the IC50 was 80 μmol/L After the treatment, cell ratio of GO and G, phases was increased (47.03±2.76 vs 56.17±1.95, t=4.68, P= 0.010), whereas the ratio of S and G2/M phases was decreased (33.07±1.86 vs 24.87±1.76, t=5.54, P = 0.010; 19.30±0.53: 17.73±0.83, t=2.75, P=0.050), and the apoptosis rate was increased (1.57±0.47:10.47±0.31, t = 27.39, P = 0.000) in a dose-dependent manner. Apoptosis with nuclear chromatin condensation, fragmentation and cell shrinkage was found by TEM. SP method showed that Celeib decreased COX-2 expression (17.48±0.34 vs 12.82±0.51,t=13.20,P =0.00). The sensitivity ratio(D0) was 1.15. FCM showed that the percentage of cells in G2/M phase was significanty more in R and D+R groups than in Ci and Cd groups (68.00±1.65,54.27±5.74,17.60±0.80,14.86±1.23, t=47.70,P=0.000; t=11.63, P=0.000), and also significantly different between R group and D + R group (t=3.99, P= 0.020). The apoptosis rate was higher in R and D + R groups than Ci and Cd groups(4.83±0.97,9.50± 1.35,1.33±0.86 and 2.28±0.42,t=4.67,P=0.010;t=8.81, P=0.000), D + R group than R group(t =4.85,P=0.010). Conclusions Celecoxib can markedly inhibit the growth and induce apoptosis in CNE-2 cells,which may depend on COX-2 pathway. Celeeoxib potently enhances the radiosensitivity of CNE-2 cells,which may due to the repair inhibit of radiation-induced DNA damage, inhibit of cell proliferation,and enhancement of cell apoptosis after irradiation.
2.Expression of SALL4 in acute myeloid leukemia and its potential clinical significance
Ye GUO ; Wei CUI ; Jingtao CUI ; Xiaodong XU ; Wei WU ; Juan DU ; Wei XIA ; Anping NI
Chinese Journal of Laboratory Medicine 2009;32(1):25-29
Objective To detect the expression of SALL4 in patients with acute myeloid leukemia (AML) and analyze its potential clinical significance. Methods Reverse transcription polymerase chain reaction and Real-time fluorescence quantitative reverse transcription polymerase chain reaction (FQ-RT-PCR) was used to examine SALLA expression in peripheral blood mononuclear cells (PBMCs) of 68 cases of AML including 36 cases in acute phase and 32 cases in remission phase, 30 healthy controls, Kasumi-1 cells and THP-1 cells. Then, flow cytometry, bone marrow smear and automated hematology analyzer were used to analyze the relationship between the SALL4 expression and blast cell counts in the bone marrow, peripheral white blood cell (WBC) counts, peripheral large unstained cell (LUC), CD34 in blast cells. Further, the change of SALL4 level during pre-chemotherapy, chemotherapy (2nd w to 3rd w) and remission were investigated in 5 AML cases. Results The level of SALL4 expression in patients with AML in acute phase [69.01 (17.20-120.28)] was 26-fold and 61-fold high compared with that in remission phase [2.64(1.35-5.41)] and in healthy control [1.14(0.50-1.62)] (Z=-6.48,-6.83,P<0.01). The level of SALL4 expression in remission phase was 2.3-fold high compared with that in healthy control (Z=-3.61 ,P<0.01). The expression level of SALL4 was decreased along with efficient chemotherapy in 5 AML cases in which SALL4 expression level was 79.74 (33.76-89.09), 7.19 (5.97-20.21) and 3.40 (1.44-15.53) during pre-chemotherapy, chemotherapy (2nd w to 3rd w) and remission, respectively. In groups of abnormal increased counts of blast cell, peripheral LUC% and CD34%, expression of SALL4 [33.82 (16.00-144.01), 30.70(23.75-72.50) and 56.25(23.79-153.81), respectively] were higher than that in groups of normal counts [2.74 (1.59-5.13), 5.71 (2.52-22.40) and 20.82 (14.03-55.12), respectively ] (Z=-4.64,-2.18,-3.66,P<0.01 or P<0.05). The expression of SALL4 in the group of increased WBC counts [89.26(23.75-154.34)] was higher than that in the group of normal WBC counts [3.86(2.03-6.01)] and the group of decreased WBC counts [6.66(2.51-17.06)] (Z=-4.91,-4.21,P<0.01). The level of SALL4 expression was positively correlated with blast cell counts in bone marrow and peripheral WBC counts (r=0.45,0.40,P<0.01). Conclusions FQ-RT-PCR method can be used successfully to detect the expression of SALL4,and the expression of SALLA may be useful to predict disease progression of AML.
3.Female sexual abnormality caused by androgen-producing adrenocortical adenoma:One case report
Xiaolin ZHANG ; Guoqing YANG ; Weijun GU ; Zhaohui LYU ; Jin DU ; Jianming BA ; Jingtao DOU ; Yiming MU
Chinese Journal of Endocrinology and Metabolism 2014;(8):673-677
Objective To describe a case of female sexual abnormality with 46, XX caused by an androgen-producing adrenocortical tumor and to explore the mechanism of abnormal androgen secretion from the tumor. Methods The tumor tissues as the experimental group were compared with the normal adrenal tissue. The LH/human chorionic gonadotropin ( hCG) receptor was determined by immunohistochemisty, the activity of 3β-hydroxysteroid dehydrogenase ( 3β-HSD ) , 17α-hydroxylase ( CYP17 ) , and 17β-hydroxysteroid oxidoreductase ( 17β-HSD ) by enzyme linked immunosorbent assay(ELISA) and the expression of mRNA of 3β-HSD2, 17β-HSDB3, CYP17, and LH/hCG receptor by real-quantitative polymerase chain reaction ( RQ-PCR ) . Results The immunohistochemisty results showed that the LH/hCG receptor was negative in the experiment group, but positive in control. The activity of 3β-HSD and CYP17 of the experiment group was higher than that in the control (P<0. 01), while the activity of 17β-HSD was lower(2 638. 798±70. 551 vs 9 148. 174±382. 836, P<0. 01) according to ELISA results. The relative contentof3β-HSD2mRNAoftheexperimentgroupwashigherthanthatinthecontrol(P<0.05),andtherelative content CYP17 mRNA of the experiment group was much higher than that in the control (P<0. 01). However, the relative content of 17β-HSDB3 mRNA and LH/hCG receptor mRNA were much lower than those in the control ( P<0. 01) by RQ-PCR. Conclusion Sexual abnormality and virilization could be caused by the excessive androgen secreted by androgen-producing adrenocortical tumor, which is an extremely rare disease. The mechanism of the secretion of androgen from the tumor remains unknown so far. It may be related to the increased activity of 3β-HSD and CYP17, but has no relationship with the expression of LH/hCG receptor.
4.Effects of celecoxib on the proliferation and apoptosis of human nasopharyngeal carcinoma cell line CNE-2
Xinhua XU ; Fang YI ; Xiangyang FU ; Daojun LI ; Qiao HUANG ; Jingtao DU
Journal of Clinical Otorhinolaryngology Head and Neck Surgery 2009;(15):682-685
Objective:To detect the effect of Celecoxib on the proliferation and apoptosis of human nasopha-ryngeal carcinoma cell line CNE-2. Method:The growth inhibition rate of CNE-2 by Celecoxib was evaluated with MTT method. Apoptosis related morphology changes were observed with transmission electron microscopy (TEM). The cell cycle and apoptosis were measured with flow cytometric method (FCM). Apoptotic index ( AI) was counted by the TDT-mediated dUTP-biotin nick end-labeling (TUNED assay. Result: The growth of CNE-2 cell was inhibited by celecoxib in a dose-and time-dependent manner. Apoptosis with nuclear chromatin condensa-tion, cell shrinkage, periplast loss and the formation of apoptotic bodies was observed with TEM. Apoptotic rates of CNE-2 cells treated with 80 and 100 μmol/L celecoxib were (10. 47±0. 18)% and (20. 17±0. 55)% respective-ly, significantly higher than those of the control group (1. 57±0. 27)% with FCM. The percentage of G_0/G_1 phase cells increased, whereas the S and G_2/M phases cells decreased in a dose-dependent manner after the treatment. TUNEL assay showed that the apoptosis ratio( AI) of CNE-2 treated with Celecoxib was higher than control group (P<0. 01). Conclusion:Celecoxib can inhibit the growth of human nasopharyngeal carcinoma cell line CNE-2 and induce the cell apoptosis, which may be related to blocking the cell cycle progress of CNE-2 cells.
5.Investigation on evaluation of pituitary function after transsphenoidal ectomy of hypophysoma
Zhichao ZHOU ; Jingtao DOU ; Zhaohui Lü ; Jianming BA ; Weijun GU ; Jin DU ; Qinghua GUO ; Juming LU
Chinese Journal of Endocrinology and Metabolism 2012;28(7):542-545
Objective To assess the pituitary function in patients with pituitary adenoma after transsphenoidal ectomy of hypophysoma.Methods Data of 106 patients with pituitary adenoma who were admitted in endocrine department and underwent the operation in PLA General Hospital from January 1993 to January 2010 were collected.Assessments of pituitary function were made before and after surgery.Results Total 23.6% and 16.0% of 106 patients underwent pituitary function evaluation by 1 week and 3 months after surgery,respectively.23.5% and 5.9% of patients with hyopituitarism before surgery underwent pituitary function evaluation by 1 week and 3 months after surgery respectively,and the respective figures in those without hypopituitarism were 23.6% and 20.8%.The incidences of new onset of hypopituitarism among 106 patients that underwent surgical procedure were 48.0% within 1 week after surgery and 35.3% by 3 months after surgery.Conclusion The rate of re-evaluation of pituitary function by 1 week and 3 months after pituitary surgery was low.Probably,many patients were missed the diagnosis of hypopituitarism after pituitary surgery.
6.Recurrent lymphocytic hypophysitis successfully treated with glucocorticoids plus azathioprine: three cases report
Guoqing YANG ; Zhaohui Lü ; Weijun GU ; Jin DU ; Qinghua GUO ; Xianling WANG ; Jianming BA ; Jingtao DOU ; Yiming MU ; Juming LU
Chinese Journal of Endocrinology and Metabolism 2011;27(1):43-46
All three cases of recurrent lymphocytic hypophysitis were female, one of them being 70-yearold postmenopausal woman.Adenohypophysis, cavernous sinus, and optic chiasm were involved in case 1,hypothalamus and neurohypophysis were invaded in case 2, and adenohypophysis and hypophyseal stalk were involved in case 3.Relapse occured 4 months after operation in case 1, then high dosage methylprednisolone pulse therapy (HDMPT) brought about a remission lasting for 14 months before second relapse set in.Relapse occurred in case 2 at 16 months after prednisone treatment was discontinued, and case 3 recurred during the period of dose reduction.All three patients were then treated with glucocorticoid plus azathioprine for 16 weeks, and good response was seen in 3 cases.During follow-up, the symptoms were relieved and significant reduction of lesion was revealed by MRI in all thee patients, and the pituitary function was resumed in case1and 3.
7.Diagnosis and treatment of 3 cases with cystic lymphangiomas of adrenal gland
Juan MIAO ; Weijun GU ; Yiming MU ; Nan JIN ; Guoqing YANG ; Qinghua GUO ; Jin DU ; Zhaohui LYU ; Jingtao DOU
Chinese Journal of Endocrinology and Metabolism 2015;(8):668-671
Objective To investigate the clinical features, diagnosis, differential diagnosis, treatment, and prognosis of adrenal lymphangioma. Methods Three cases of adrenal lymphangioma were reported, and the clinical features, treatment and prognosis were analyzed. Results Three cases were incidentally discovered, laboratory tests and endocrine hormone examinations were normal, CT or MRI showed lesions with low density, no reinforced or mild enhancement. All 3 cases underwent laparoscopic adrenalectomy, postoperative pathology supported the diagnosis of adrenal lymphangioma. They were followed up for 8-months, 1-year, and 4-years respectively, with no recurrence. Conclusions Adrenal lymphangioma is a rare benign adrenal leison, with no typical clinical manifestations. Preoperative diagnosis depends on imaging examinations. Histopathological examination is essential in making final diagnosis. Surgery is the preferred treatment option. The prognosis is relatively good.
8.Clinical Value of Captopril Test for Primary Aldosteronism Diagnosis
Lixue WANG ; Yiming MU ; Jianming BA ; Jingtao DOU ; Zhaohui LU ; Xianling WANG ; Jin DU ; Guoqing YANG ; Juming LU
Chinese Circulation Journal 2016;31(8):772-774
Objective: To evaluate the clinical value of Captopril test for diagnosing primary aldosteronism (PA) and to calculate the best cut-off point for PA diagnosis. Methods: We retrospectively analyzed 96 PA patients with conifrmed diagnosis by clinical situation, laboratory test and auxiliary examination in our hospital from 1994-06 to 2012-05, and meanwhile, studied 45 highly suspicious PA patients with final exclusion by confirmed diagnosis of primary hypertension (PH). All patients received the in-hospital Captopril test, the area under the curve of receiver operating characteristic (AUCROC) was applied to evaluate plasma aldosterone level and the ratio of aldosterone/renin after Captopril test and to obtain the best cut-off point with the corresponding sensitivity and speciifcity for PA diagnosis. Results: At 1h and 2h after Captopril test, AUCROC for plasma levels of aldosterone were 0.831 and 0.818, the ratios of aldosterone/rennin were 0.909 and 0.922 respectively. At 1h after Captopril test, the cut-off point of aldosterone level was 544.95 pmol/L and the diagnostic sensitivity was 70%, speciifcity was 90.7%; at 2h after Captopril test, the cut-off point of aldosterone level was 466.8 pmol/L and the diagnostic sensitivity was 69.8%, speciifcity was 70.5%. At 1h after Captopril test, the ratio of aldosterone/rennin was 34.6 [ng/dl: μg/(ml·h)] with the sensitivity at 78.3% and speciifcity at 88.4%. At 2h after Captopril test, the maximum AUCROC for the ratio of aldosterone/rennin was obtained, when cut-off point of aldosterone level was 42.2[ng/dl: μg/(ml·h)] , the diagnostic sensitivity was 76.7%, speciifcity was 95.3%. Conclusion: At 1h and 2h after Captopril test, plasma aldosterone level and the ratio of aldosterone/rennin had been valuable for PA diagnosis, the maximum diagnostic value could be obtained at 2h after Captopril test.
9.Diagnostic Value of Saline Load Test in Patients With Primary Aldosteronism
Lixue WANG ; Yiming MU ; Jianming BA ; Jingtao DOU ; Chaohui LV ; Xianling WANG ; Jin DU ; Gouqing YANG ; Juming LU
Chinese Circulation Journal 2016;31(6):604-605
Objective: To evaluate the diagnostic value of saline infusion test (SIT) in patients with primary aldosteronism (PHA). Methods: A total of 116 patients with PHA or essential hypertension (EH) treated in our hospital from 1994-06 to 2013-05 were retrospectively studied. The patients were divided into 2 groups: PHA group,n=72 and EH group, the patients with excluded PHA,n=44. post-SIT plasma levels of aldosterone and post-SIT ratio of aldosterone/renin activity were evaluated by ROC curve in order to analyze the diagnostic capability and the best diagnostic cut-off point. Results: The area under curve (AUC) by ROC for post-SIT aldosterone level was 0.759, the sensitivity and speciifcity were 74.6% and 63.6% respectively; AUC for post-SIT ratio of aldosterone/renin activity was 0.899, the sensitivity and speciifcity were 83.6% and 88.6% with the best diagnostic cut-off point at 111 [ng/dl:ng/(ml?h)]. Conclusion: Post-SIT plasma level of aldosterone and post-SIT ratio of aldosterone/renin activity had the diagnostic value of PHA; post-SIT ratio of aldosterone/renin activity had the higher diagnostic value of PHA.
10.A study on clinical features of ACTH-independent bilateral macronodular adrenal hyperplasia
Qian ZHANG ; Jingtao DOU ; Weijun GU ; Jin DU ; Xinnling WANG ; Guoqing YANG ; Qinghua GUO ; Lijuan YANG ; Zhaohui Lü ; Jianming BA ; Yiming MU ; Juming LU ; Changyu PAN
Chinese Journal of Endocrinology and Metabolism 2011;27(11):892-896
Objective To study the clinical features of patients with ACTH-independent bilateral macronodular adrenal hyperplasia(AIMAH).Methods Eighteen cases with AIMAH diagnosed in Chinese PLA General Hospital from 1998 to 2010 were analyzed retrospectively.Results ( 1 ) AIMAH was equally distributed between genders,mean age at diagnosis was ( 51.4 ± 10.7 ) years,and average course was ( 9.9:t:2.7 ) years.( 2 )Most patients visited hospital because of adrenal incidentaloma; hypertension and impaired glucose tolerance/diabetes mellitius were the most common clinical presentations; typical signs of Cushing′s syndrome (CS) such as moon face and central obesity were less frequent.( 3 ) All cases′ laboratory findings met the diagnostic criteria of ACTH-independent CS,some cases with the features of subclinical CS.24 h dynamic plasma ACTH and cortisol monitoring had the advantage of revealing the features of subclinical CS for some AIMAH cases; there was remarkable elevation of plasma cortisol after exogenous ACTH stimulation in AIMAH patients.(4) Bilateral adrenal macronodular changes could be visualized on CT scan.( 5 ) Pathology identified bilateral macronodular adrenal hyperplasia.( 6 ) Bilateral adrenalectomy was the most useful treatment.Conclusions AIMAH is an infrequent cause of CS,and some patients present subclinical CS.Its marked clinical feature is ACTH-independent bilateral macronodular adrenal hyperplasia.