1.Effect of DNA methyltransferase 1 gene silencing on proliferation and apoptosis of HeLa cells
Xianlan ZHAO ; Yanling RAO ; Yuhuan QIAO ; Huili ZHANG
Chinese Journal of Obstetrics and Gynecology 2009;44(7):533-537
Objective To observe the effect of DNA methyltransferase 1 (DNMT1 ) gene silencing by RNA interfering technology on the proliferation and apoptosis of HeLa cells. Methods Recombinant plasmid pshRNA-DNMT1-A, B and C were respectively transfected into HeLa cells by lipofectamine 2000, while cells transfected plasmid vector pSilencer3.1-HI and cells untreated as control groups. RT-PCR was adopted to select the recombinant plasmid which showed the most optimal inhibition effect. RT-PCR and western blotting was used to detected the mRNA and protein expression of DNMT1 in HeLa cells transfected for 24, 48 and 72 hours. Cell counting kit-8 (CCK-8 ) assay was used to investigate the proliferation of the HeLa cells after transfection, while apoptosis was detected by flowcytometry(FCM ) method. Results Three DNMT1-targeted short hairpin RNA (shRNA) A,B and C were successfully inserted into the plasmid vector PShRNA, and the coding sequences of the obtained shRNA were consistent with the designed fragments. The results indicated that both recombinant plasmid pshRNA-DNMT1-A and B could effectively knock down the expression of DNMT1 gene in human cervical cancer cells, of which pshRNA-DNMTI-B was the better choice. While no effect of pshRNA-DNMTI-C was seen. BT-PCR results showed that the relative mRNA expression of DNMT1 gene in Helm cells transfected with pshRNA-DNMT1 for 24, 48 and 72 hours were 0.406±0.057,0.191±0.036 and 0.104±0.015, which were significantly lower than that in Helm cells transfected by empty vector and non-transfected cells (0.520±0.020, 0.537±0.041, respectively, P < 0.05 ). The western blotting analysis manifested that the relative expression of DNMT1 protein of Helm cells transfected by pshRNA-DNMT1 for 24, 48 and 72 hours were 0.197±0.024, 0.075±0.015, 0.040± 0.013, which were significantly lower than that in transfected cells by empty vector and non-transfected cells (0.273±0.010, 0.283±0.016, respectively, P <0.05). The CCK-8 results showed that the cell survival rates of HeLa cells transfected by pshRNA-DNMT1 for 24, 48, 72, 96 and 120 hours were 70.8%, 64.8%, 51.6%, 45.3% and 38.0%, there were statistically different compared with cells transfected by empty vector and non-transfected cells at different time-points (P < 0.01 ). The results of FCM indicated that the apoptesis rate of HeLa cells trandected with pshRNA-DNMTI for 24, 48 and 72 hours were (17.7± 1.3 ) %, (35.3±1.3 ) %, (47.6±1.6 ) %, which were significantly higher than empty vector transfected cells and non-transfected cells [(4.9±0.5 ) %, (5.1±0.7 ) %, respectively, P < 0.05]. Conclusions DNMT1 can be successfully silenced by RNA interfering in cervical Helm cells. Downregulation of DNMT1 can inhibit cervical cancer cells proliferation and induce cell apoptosis.
2.The Effects of Carboxymethylated Starch on Childhood Extrinsic Asthma and Its Immunological Mechanism
Yuhuan ZHAO ; Zhijia ZHOU ; Lihua HUANG ; Kunfeng WU ; Jinjin JIANG
Academic Journal of Second Military Medical University 1985;0(06):-
45 cases of childhood extrinsic asthma (CEA) were divided into the treatment group with carboxymethykited starch (CMS) (n=30) and the control group (n=15). The total effective rate of the former was 63% (P
3.Quality control in maternal serum prenatal screening in the second trimester
Fengxian SHEN ; Li ZHAO ; Shiming LYU ; Yuhuan SHEN
Chinese Journal of Laboratory Medicine 2014;(11):870-875
Objective To analyze the features of quality management of maternal serum prenatal screening for Down syndrome in the second trimester and investigate the methods to establish and improve quality control system.Methods Prenatal screening and clinical data were collected for statistical analysis from 17 prenatal screening centers in Zhejiang Province from 2011 to 2012.The impacts of maternal age , weight, gestational age , specimen collection , the median values of serum markers selection and different analysis software on prenatal screening results for Down syndrome were analyzed respectively .Results Totally 417 347 and 463 846 prenatal screening cases were collected in 2011 and 2012 respectively.The maternal age of the entire province presented as a Gaussian distribution.The age distributions of different screening institutions were inconsistent , while maternal weight distributions were similar.The approach to determine gestational age was significantly different among screening institutions.Median MoM of the samples from the blood collection points was different from those of the main screening center.Median MoM-freeβ-hCG was positive bias , particularly the specimens from the blood collection points.After strengthening the control of temperature and time during samples transportation , Median MoM-free β-hCG of specimens from blood collection points became nearly equal to those of main screening center .Use of local median made the median MoM value closer to 1.0 in the centers with positive or negtive bias of the markers , which had little effect in detection rate and decreased the false positive rate and screening positive rate.Six more positive cases were detected by the Lifecycle software instead of 2T software in 213 followed-up positive cases, which means the Lifecycle software can improve the detection rate.Conclusions The impact of maternal age, weight, gestational age, specimen collection and other factors on screening performance should be taken into consideration when it evaluates the screening quality of different laboratory .The use of local medians is necessary in prenatal screening.Optimization of screening analysis software can reduce false negative rate.
4.Study of correlation between polymorphism of HLA-DRB allele and susceptivity of atopic dermatitis
Hongli ZHAO ; Xueqian WANG ; Yuhuan ZHANG ; Xiaohui SUN ;
Chinese Journal of Laboratory Medicine 2000;0(06):-
Objective To look for susceptible and resistant HLA gene of Atopic Dermatitis(AD), analyze single strand conformation polymorphism of the susceptible gene and observe whether the base sequence was different from healthy Method Polymerase chain reaction sequence specific primer(PCR SSP)was used to classify the HLA DRB allele of 122 patients and 80 controls PCR single strand conformation polymorphism(SSCP) was used to analyse the susceptible gene Results The DR 3 frequency of extrinic type of “pure” AD(EAD)group and complicated group was higher than that of the control group ( RR =5 27 and 3 59 respectively P 0 05). Conclusion HLA DR 3 was the susceptible gene of AD, while HLA DR 6 was the resistant gene of AD It indicated that people with HLA DR 3 were reliable to AD, while people with HLA DR 6 were not But there was no base difference between these people and the healthy in HLA DR 3
5.Application research of dry powder from stone of Garcinia oblongifolia in south of the Five Ridges in treatment of refractory Ⅲ and Ⅳ stage bedsore
Jun YUAN ; Xuejin ZHAO ; Guifen ZHANG ; Yuhuan WU ; Yanhui ZENG ; Zhizhu WU
Chinese Journal of Practical Nursing 2013;29(19):27-28
Objective To discuss the effect of dry powder from stone of Garcinia oblongifolia in south of the Five Ridges on refractory Ⅲ and Ⅳ stage bedsore.Methods 120 cases of patients with refractory Ⅲ and Ⅳ stage pressure sore were randomly divided into the treatment group and the control group with 60 cases in each group.The treatment group applied dry powder from stone of Garcinia oblongifolia in south of the Five Ridges,while the control group used conventional infrared irradiation plus chloramphenicol powder treatment method.The treatment effect was compared between two groups.Results There were significnat differences in clinical efficacy,pain relief and cure time between the treatment group and the control group,and the treatment group acquired better effect than the control group.Conclusions The clinical efficacy,pain relief and healing time of the treatment group in the treatment of refractory Ⅲ and Ⅳ stage pressure ulcer was superior to the control group,and without skin lesions and human adverse reaction,and with the advantages of simple operation,quick,good curative effect,no pain to the patient,easy to accept and no toxic side effect.
6.Clinical observation of pulmonary function in patients of be severe secondary hyperparathyroidism caused by uremia
Yuhuan SONG ; Yiping WANG ; Lianxiang QIN ; Na ZHAO ; Yuefei XIAO ; Ling ZHANG ; Li YAO
Military Medical Sciences 2014;(10):819-822
Objectives To evaluate the lung function in patients of severe secondary hyperparathyroidism caused by uremia and to analyze related factors.Methods The pulmonary function of 70 patients with uremia ( 40 patients with severe secondary hyperparathyroidism, 30 MHD patients without SHPT) and 30 healthy people was determined.Correlative analysis was performed between parameters such as pulmonary function indexes and clinical or laboratory parameters. Results Lung function was significantly different between patients with uremia and healthy subjects(P<0.05, respectively). A number of lung function indicators were negatively correlated with iPTH,C reactive protein( CRP) ,dialysis age but were positively correlated with hemoglobin (Hb) and albumin(Alb) (P <0.05).Conclusion Patients with SHPT show impaired lung function.IPTH, dialysis age, Hb, Alb and CRP can be used as important indicators of lung function.
7.The study of distinguishing solid solitary pulmonary nodule with diffusion weighted imaging
Jinjun REN ; Baohong ZHAO ; Xuerong ZI ; Yuhuan ZHANG ; Hongwei CHI ; Shuping WANG ; Qian XU ; Yanfang CHEN
Journal of Practical Radiology 2015;(6):925-928
Objective To evaluate the value of diffusion weighted imaging (DWI)in distinguishing the solid solitary pulmonary nodule (SPN).Methods 42 patients with SPN (malignant in 25 and benign in 1 7)who were confirmed by operation,biopsy or follow up after treatment underwent routine chest T1 WI,T2 WI and DWI.The b values were chosen as 300,500,800 and 1 000 s/mm2 ,and the corresponding apparent diffusion coefficient (ADC)values and the signal intensity (SI)were respectively measured.Results The ADC values and SI of benign and malignant SPNs were gradually reduced with increasing b value.The ADC value between benign and malignant SPNs was statistically significant with b value of 500 s/mm2 (P 500 =0.03 <0.05 ),meanwhile the SI was statistically significant with b values from 300 to 1000 s/mm2 (P 300 <0.001,P 500 =0.03 <0.05,P 800 =0.01 <0.05, P 1 000 =0.02<0.05).Conclusion Both SI and ADC value of DWI play important role in distinguishing benign and malignant SPNs, and the diagnostic efficiency of SI is superior to ADC value.
8.Association of rs3660 Single Nucleotide Polymorphisms with Non-Hodgkin's Lymphoma
Guimin ZHAO ; Yuhuan GAO ; Lihong LIU ; Xiaolin WU ; Zhe GAO ; Lanping DIAO
Journal of China Medical University 2017;46(4):321-325
Objective To evaluate the effect of single-nucleotide polymorphisms at the miRNA binding site rs3660 in the 3'-untranslated region of the KRT81 gene (miR-SNPs) on the cancer risk and clinical prognosis of non-Hodgkin's lymphomas (NHL).Methods The single-nucleo-tide polymorphisms of rs3660 was genotyped with ligation detection reaction method.The association of rs3660 with NHL survival was calculated with log-rank test using Kaplan-Meier method.Multivariate survival analysis was performed using a Cox proportional hazards model.Results The rs3660 genotype distribution difference was not statistically significant between the case and control group (P =0.50).Patients carrying the rs3660 CG/CC genotype exhibited a significantly longer survival time than patients carrying the GG genotype (P =0.012).In addition,rs3660 was associated independently with the survival of NHL patients in multivariate analysis (RR=0.589,95% CI:0.415-0.832,P =0.004).The association of this miR-SNP with NHL survival was further confirmed in the peripheral T cell lymphoma (PTCL) subtype.Conclusion Our results indicate that KRT81 rs3660 GG type is an independent prognostic marker in NHL.
10.Interpretation of ICRU report No.89-radiation physics
Hongfu ZHAO ; Dongmei HAN ; Guanghui CHENG ; Yuhuan TANG
Chinese Journal of Radiation Oncology 2019;28(1):74-77
In this article,the content of radiation physics in the ICRU report No.89 were interpreted in details,aiming to provide reference in the radiation physics for radiologists performing brachytherapy for cervical cancer patients.