3.Effects and mechanisms of 2-methoxyestradiol on the expression of apoptosis-related genes in K562 leukemia cells
Dongliang LI ; Jing ZHANG ; Ling PAN ; Lanfen HOU ; Zhiwei WANG ; Xiao GUO
Journal of Leukemia & Lymphoma 2009;18(6):323-326,330
Objective To investigate the effects of 2-methoxyestradiol( 2-ME) on apoptosis of K562 cells and its mechanisms. Methods The K562 cells were cultured and divided into three groups. The control group: K562 cells were cultured without 2-ME treatment. The experimental group: K562 cells were cultured in the medium containing different concentrations of 2-ME (1, 2, 4, 8 and 16 μmol/L) for 36 h. The negative control group: K562 cells were replaced by water without RNase in the medium containing different concentrations of 2-ME for 36 h. The apoptosis rate, the protein and its mRNA expression of Caspase-3 and XIAP, the activity of superoxide dismutase (SOD) and reactive oxygen species (ROS) of K562 cells wasdetected by TUNEL, flow cytometry (FCM), half-quantitative RT-PCR and xanthenes oxidized enzyme assay,respectively. Results After treated with 2-ME at different concentrations for 36 hours, in the specified concentration range, 2-ME induced apoptosis of K562 cells in a concentration-dependent manner. The possible functional mechanism of 2-ME was to up-regulate Caspase-3 but down-regulate XIAP mRNA expression, and increase ROS activity but decrease SOD activity. Conclusion 2-ME can induce apoptosis of K562 cells in a concentration-dependent manner and indicate its promising potential in the treatment of chronic myeloid leukemia(CML) patients.
4.Cloning and expression of the key enzyme hyoscyamine 6 beta-hydroxylase gene (DaH6H) in scopolamine biosynthesis of Datura arborea.
Wei QIANG ; Yan-ling HOU ; Xiao LI ; Ke XIA ; Zhi-hua LIAO
Acta Pharmaceutica Sinica 2015;50(10):1346-1355
Hyoscyamine 6 beta-hydroxylase (H6H) is the last rate-limiting enzyme directly catalyzing the formation of scopolamine in tropane alkaloids (TAs) biosynthesis pathway. It is the primary target gene in the genetic modification of TAs metabolic pathway. Full-length cDNA and gDNA sequences of a novel H6H gene were cloned from Datura arborea (DaH6H, GenBank accession numbers for cDNA and gDNA are KR006981 and KR006983, respectively). Nucleotide sequence analysis reveals an open reading frame of 1375 bp encoding 347 amino acids in the cDNA of DaH6H, while the gDNA of DaH6H contains four exons and three introns, with the highest similarity to the gDNA of H6H from D. stramonium. DaH6H also exhibited the most identity of 90.5% with DsH6H in amino acids and harbored conserved 2-oxoglutarate binding motif and two iron binding motifs. The expression level of DaH6H was highest in the mature leaf, followed by the secondary root, and with no expression in the primary root based on qPCR analysis. Its expression was inhibited by MeJA. DaH6H was expressed in E. coli and a 39 kD recombinant protein was detected in SDS-PAGE. Comparison of the contents of scopolamine and hyoscyamine in various TAs-producing plants revealed that D. arborea was one of the rare scopolamine predominant plants. Cloning of DaH6H gene will allow more research in the molecular regulatory mechanism of TAs biosynthesis in distinct plants and provide a new candidate gene for scopolamine metabolic engineering.
Cloning, Molecular
;
DNA, Complementary
;
Datura
;
enzymology
;
genetics
;
Escherichia coli
;
Hyoscyamine
;
chemistry
;
Mixed Function Oxygenases
;
genetics
;
Plant Leaves
;
enzymology
;
Plant Roots
;
enzymology
;
Recombinant Proteins
;
genetics
;
Scopolamine Hydrobromide
;
chemistry
5.The effects of rapamycin on the proliferation and apoptosis of human retinal pigment epithelium cells in vitro
Min, HOU ; Hong-ling, LIU ; Zeng-wei, YIN ; Xiao-mei, ZHANG
Chinese Journal of Experimental Ophthalmology 2011;29(10):879-883
Background Rapamycin(RAPA)is a specific inhibitor of the mammalian target of rapamycin (mTOR).Researches showed that RAPA inhibits the proliferation of lens epithelium cells(LECs)and tumor cells and induces apoptosis of tumor cells.To investigate whether rapamycin has the inhibitory effect on retinal pigment epithelium(RPE)cells is very important for the prevention and management of proliferative vitreoretinopathy (PVR).Objective This study was to investigate the effects of RAPA on the proliferation and apoptosis of human RPE cells in vitro.Methods Human RPE cells(D407 strain)were cultured and passaged and then divided into regular culture group(blank control group),DMSO control group(0.1‰ DMSO +regular culture),and different concentrations RAPA-treatment groups(5,10,20,40,80,160,320 nmol/L).The proliferation(A490)of human RPE cells was detected using MTT,and the inhibitory rates of RAPA on the proliferation of RPE cells were calculated and compared among different groups at 12,24 and 48 hours.The apoptosis rates of the cells were analyzed among various groups by Hoechst staining after 12,24,48 hours.Results The inhibitory rates of RAPA on RPE cells were significantly different among various groups(F=484.451,P<0.01)and evidently elevated in 20-320 nmol/L RAPA groups compared with DMSO control group(P < 0.01).The inhibition of RAPA on the cells was considerably enhanced as the lapse of time(F=232.262,P<0.01)with more dominant effects in 24 and 48 hours compared to 12 hours after addition of RAPA(P<0.05-0.01).Compared with blank control group and DMSO control group,the apoptotic rates of the cells were evidently increased in 12,24,48 hours in 10 nmol/L RAPA group(all P<0.05),and higher cellular apoptotic rates were found in 20-320 nmol/L RAPA groups(all P<0.01).The alteration of cellular apoptotic rate showed a gradually incremental trend as the acting time of RAPA(F =625.584,P<0.01).Karyorrhexis and mass-like density staining and chromatin substance were seen in RPE cells under the fluorescence microscope in ≥ 10 nmoL/L RAPA groups.Conclusions RAPA suppresses the proliferation and induces the apoptosis of human RPE cells in concentration-and time-dependent manner in vitro.
6.Independent Relationship between Body Mass Index and LH Peak Value of GnRH Stimulation Test in ICPP Girls: A Cross-sectional Study
ZHAO YUE ; HOU LING ; GAO HONG-JIE ; ZHAN DI ; ZHANG CAI ; LUO XIAO-PING
Journal of Huazhong University of Science and Technology (Medical Sciences) 2017;37(4):556-562
The effect of obesity on idiopathic central precocious puberty (ICPP) girls is still under discussion.The relationship between body mass index (BMI) and sexual hormone levels of gonadotropin-releasing hormone (GnRH) stimulation test in ICPP girls is controversial and the underlying mechanism is unclear.This study aims to further explore the independent effect of excess adiposity on peak luteinizing hormone (LH) level of stimulation test in ICPP girls and the role of other related factors.A retrospective cross-sectional study was performed on 618 girls diagnosed as having ICPP,including 355 cases of normal weight,99 cases of overweight and 164 cases of obese.The results showed that obese group had more progressed Tanner stage and no significant difference (P=0.28) in LH peak was found as basal LH value was used as a covariate.The obese group had higher total testosterone (TT),adrenocorticotrophic hormone (ACTH),17-α hydroxyprogesterone (17-αOHP) and androstendione (AN),with significantly increased fasting insulin (FIN) and homeostasis model of assessment for insulin resistance index (HOMA-IR).Stratified analysis showed inconsistency of the relationship between BMI-standard deviation score (BMI-SDS) and LH peak in different Tanner stages (P for interaction=0.017).Further smoothing plot showed linear and non-linear relationship between BMI-SDS and LH peak in three Tanner stages.Then linear regression model was used to analyze the relationship between BMI-SDS and LH peak in different Tanner stages,with and without different confounding factors being adjusted.In B2 stage,BMI-SDS was negatively associated with LH peak.In B3 stage,when BMI-SDS <1.5,as BMI-SDS increased,the level of LH peak decreased (model Ⅰ:β=-1.8,95% CI=-4.7 to 1.1,P=0.214).When BMI-SDS ≥1.5,BMI-SDS was significantly positively associated with LH peak (model Ⅰ:β=4.5,95% CI=1.7 to 7.4,P=0.002).In B4 stage,when BMI-SDS <1.5,BMI-SDS was negatively associated with LH peak (model Ⅰ:β=-11.6,95% CI=-22.7 to-4.5,P=0.049).When BMI-SDS ≥1.5,BMI-SDS was positively associated with LH peak (model Ⅰ:β=-4.2,95% CI=-3.3 to 11.7,P=0.28).It is concluded that there is an independent correlation between BMI-SDS and LH peak of stimulation test in ICPP girls,their relationships are different in different Tanner stages,and the effect of BMI-SDS can be affected by adrenal androgens,estradiol and glucose metabolism parameters.
7.Senile dementia: differentiation of syndromes according to meridians based on the theory of "cerebral collaterals injury by toxin".
Li ZHOU ; Yun-Ling ZHANG ; Xiao-Bing HOU ; Hui HU
Chinese Acupuncture & Moxibustion 2012;32(11):1031-1034
The pathogenesis and treatments based on meridian differentiation of senile dementia are discussed through analyses and researches on the theory of "cerebral collaterals injury by toxins" and "collateral diseases". The symptoms of "Cerebral collaterals injury by toxins" are preliminary characterized by toxins and blood stasis occluding brain collaterals. "Cerebral collateral injury by toxins" and "Governor Vessel occlusion by blood stasis" are taken as the major pathogeneses of senile dementia. And the treatment should be focused on clearing the collaterals. Clearance acting as reinforcing as well as to clear and modify the Governor Vessel are taken as crucial sections in the treatment of senile dementia based on meridian differentiation. It is also the application of acupuncture-moxibustion intervention in senile dementia based on the theory of "cerebral collateral injury by toxins", which expands the application of the theory concerning "collateral diseases" in disease prevention and treatment with acupuncture-moxibustion.
Acupuncture Therapy
;
Alzheimer Disease
;
diagnosis
;
metabolism
;
therapy
;
Brain
;
drug effects
;
metabolism
;
Diagnosis, Differential
;
Humans
;
Meridians
;
Toxins, Biological
;
metabolism
;
pharmacology
8.Abdominal aortic calcification detected by lateral lumbar X-ray in maintenance hemodialysis patients
Zijin CHEN ; Xiaonong CHEN ; Xiaobo MA ; Liang HOU ; Bei DING ; Huawei LING ; Xiao LI ; Hong REN ; Nan CHEN
Chinese Journal of Nephrology 2012;28(8):606-610
Objective To explore the incidence and associated factors of aortic artery calcification (AAC) by lateral lumbar X-ray score in maintenance hemodialysis (MHD) patients.Mehtods A total of 155 MHD patients with complete clinical data in our hospital were enrolled in the study.Lateral lumbar X-ray score of the abdominal aorta was used to determine AAC in MHD patients.Results Aortic calcification was most severe in front of the fourth lumbar segment and ameliorated in higher lumbar levels.63.63% of MHD patients presented visible calcification in the abdominal aorta,and 28.39% had severe calcification with more than three segments.Age (OR=1.094,P<0.01),dialysis vintage (OR=1.013,P=0.022),triglyceride (OR=1.261,P=0.030) and phosphate level (OR=1.324,P=0.023) were risk factors of abdominal aorta calcification,however serum albumin level (OR =0.239,P=0.013) was protect factor of aortic calcification.Conclusions Incidence of AAC is quite high in MHD patients and associated with increasing of age,duration of hemodialysis,serum triglyceride,phosphate level and plasma albumin.The semi-quantitative X-ray method of determining vascular calcification is less expensive and may be widely available clinically.
9.Clinical characteristics of Wolfram syndrome.
Ling HOU ; Han-Hua LIN ; Li WU ; Xiao-Ping LUO
Chinese Journal of Contemporary Pediatrics 2009;11(2):113-115
OBJECTIVEWolfram syndrome (WFS) is a rare, autosomal recessive inherited disease characterized by various clinical manifestations. The aim of this study was to investigate clinical characteristics of WFS.
METHODSOne case of WFS was reported. Combined with the clinical data of 8 cases of WFS which had been reported in China between 1994 and 2007, the clinical characteristics of WFS were reviewed.
RESULTSInsulin-dependent diabetes mellitus as the earliest manifestation was found in all of the 9 patients, with a median onset age of 5.0 years. Optic atrophy occurred in 8 patients (onset age: 8.5 years), diabetes insipidus in 7 patients (onset age: 8.5 years) and deafness in 7 patients (onset age: 9.8 years). Short stature was found in 6 patients and hydroureteronephrosis in 4 patients.
CONCLUSIONSInsulin-dependent diabetes mellitus was the first presentation in children with WFS. Optic atrophy, diabetes insipidus and deafness were common complications, with a various onset age.
Child ; Humans ; Male ; Wolfram Syndrome ; complications ; diagnosis ; therapy
10.Totic Implantable Collamer Lens for extreme high myopic astigmatism
Zhi-gao, ZUO ; Li, YIN ; Su-bing, LIU ; Ying, HOU ; Xiao-li, NIE ; Zhi-bin, MAI ; Hong-ling, JIANG
Chinese Journal of Experimental Ophthalmology 2011;29(5):460-463
Background In recent years,with the contiunous progress of the refractive surgery,the operation skill of phakic intraocular lens(PIOL)implantation for correcting extreme high myopia,astigmatism,farsightedness have made greater progression,and its security,effectiveness in clinical attract much more attention. Objective This study was to evaluate the efficacy,safety and stability of Toric Implantable Collamer Lens(TICL)for extreme high myopic astigmatism. Methods This retrospective case series included 33 eyes of 27 patients from May 2008 to February 2009.A TICL was intraocularly implanted via a 3 mm clear corneal incision after paraocular anesthesia.Patients were examined preoperatively and followed-up at 1 day,1 week,1 month,3,6,12 and 18 months postoperatively.The examinations included uncorrected visual acuity,best corrected visual acuity(BCVA),slit lamp examination,refraction,intraocular pressure,endothelial cell morphometry,etc.The written informed consent was obtained from each patient before any medical procedure. Results The uncorrected visual acuity in 96.97% eyes was equal or improved after operation in comparison with BCVA of preoperation.The spherical refraction was within-1.00 D-+0.25 D.The cylinder refraction was within-1.00 D-0 D.The axial deviation of TICL within 10 degree was 93.94%(31/33).No significant differences were found in the intraocular pressure and endothelial cell morphometry between preoperation and postoperation(intraocular pressure:F=3.35,P=5.49;endothelial cell morphometry:t=1.835,P=0.082).The visual acuity and refraction were stable during the follow-up.Astigmatic axial rotation required surgical intervention on one eye.One eye occurred high intraocular pressure because of bigger TICL diameter.The intraocular pressure returned to normal after TICL was exchanged.No cataract occurred during the follow-up duration. Conclusion TICL implantation appears to be an effective,safe and reliable method for extreme high myopic astigmatism.