1.Effect of APRIL siRNA in Nude Mice with SW480
Xiaojuan REN ; Jingchun WANG ; Haidan CHU ; Hui CONG
Journal of Modern Laboratory Medicine 2014;(6):55-58
Objective To investigate the role of APRIL in SW480 cell line.Methods The CRC model was established in the nude mice,all the mice were divided into 3 groups,the mice were separately treated with APRIL siRNA,pGC-vector and PBS solution.The APRIL mRNA was detected by RT-PCR and the APRIL protein was surveyed by the way of immunohis-tochemistry (IHC).The proteins of TIMP-3,Syndecan-1and MMP-9 also were assessed by IHC.Results ①Tumor mass in the group of nude mice injected with PBS (2.15±0.30 g)was significantly higher than the injection APRIL siRNA group (0.95±0.15 g,P<0.05),and was not statistically significant (P>0.05)compared with the injection of empty vector group (2.20±0.25 g).②APRIL mRNA/18S rRNA ratio (2.48±0.25)in the group of mice injected with PBS was signifi-cantly higher than the injection APRIL siRNA group (0.39±0.15,P<0.05),and was not statistically significant (P>0.05)compared with the injection of empty vector group (2.51±0.30).③SW480 cells injected with APRIL siRNA signifi-cantly inhibited invasion and metastasis.TIMP-3 Allred scores in three groups were 7.70±0.35,1.10±0.16 and 1.15± 0.12,Syndecan-1 protein was 7.80±0.30,1.05±0.20 and 1.10±0.22 MMP-9 protein was 1.20 ±0.10,8.00±0.25 and 8.20±0.20,respectively.Conlusion APRIL was closely connected with the growth and metabasis of CRC.
2.Differences of genome-wide methylation level and methylated genes be-tween nasopharyngeal carcinoma cells in same genetic background but different radiation resistance
Haidan LUO ; Donghui WANG ; Weida LIN ; Bojin SU ; Cunyou ZHAO ; Huiling YANG
Chinese Journal of Pathophysiology 2015;(7):1300-1304
[ ABSTRACT] AIM:To compare the differences of the genome-wide methylation levels and methylated regions be-tween nasopharyngeal carcinoma ( NPC) cells in the same genetic background but different radiation resistance ( CNE-2 cells and CNE-2R cells).METHODS:Using the method which was developed by Doctor Zhao Cun-you, based on using methyl-sensitive restriction enzyme to measure the genome-wide methylation levels.In addition, MeDIP-Seq was used to analyze the methylated regions in 6 gene functional elements, including the upstream 2k sequence, 5’ UTR, coding se-quence, intron, 3’UTR and downstream 2k sequence, between CNE-2 cells and CNE-2R cells.RESULTS:The genome-wide methylation level was approximately 30%lower in CNE-2R cells than that in CNE-2 cells.No obvious difference on the amount of genes and the coverage of the peak in the 6 gene functional elements was observed.However, the methylation pattern of plentiful genes had altered in the gene function elements.CONCLUSION:The genome-wide methylation levels and methylated regions between NPC cells in the same genetic background but different radiation resistance were quite dif-ferent, indicating that the DNA methylation may be associated with NPC radioresistance.
3.Preparation of monoclonal antibody against LRR-WSC domain of polycystin-1 and distribution of polycystin-1 in kidney tissues and kidney cell lines
Haidan ZHAO ; Changlin MEI ; Xuefei SHEN ; Tianmei SUN ; Shuzhong ZHANG ; Wenjing WANG ; Yume WU ; Ji SONG
Chinese Journal of Laboratory Medicine 2003;0(08):-
Objective To prepare and identify monoclonal antibody against LRR-WSC domain of polycystin-1 and to investigate the distribution of polycystin-1 in kidney tissues and kidney cell lines. Methods BALB/c mice were immunized with fusion protein PC1-e of polycystin-1 LRR-WSC domain. The splenocytes were fused with myeloma cells by PEG 4000 and the hybridomas were selected in HAT medium. The hybridoma clones secreting antibodies against polycystin-1 LRR-WSC domain were detected by enzyme-linked immunosorbent assay ( ELISA) and cloned by limiting dilution. The specificity of anti-polycystin-1 LRR-WSC domain monoclonal antibody from hybridoma was verified by ELISA and Western blot. The distribution of polycystin-1 in tissues and cells was detected by immunohistochemical method. Results One cell line of hybridoma secreting monoclonal antibody against polycystin-1 was established. Western blot analysis showed that the monoclonal antibody reacted strongly and specifically to polycystin-1 LRR-WSC domain. Distribution of polycystin-1 in fetal kidney was localized in tubular epithelium. In normal adult kidney tissues, our study showed that polycystin-1 was mainly expressed in the medullary collecting ducts and distal convoluted tubules. Positive staining was also found in the majority of cyst-lining epithelial ceEs of cystic tissue from autosomal dominant polycystic kidney disease ( ADPKD) patients. Expressions of polycystin-1 were found in either ADPKD cyst-lining epithelia cell line and LLC-PK1, clearly plasma membrane and intracytoplasmic staining of polycystin-1 were observed. Conclusion Specific monoclonal antibody against polycystin-1 LRR-WSC domain were obtained. The antibody is important to researching the mechanism of ADPKD. The distribution of polycystin-1 in kidney tissues and cells show that polycystin-1 was important in tubular elongation and the maintenance of tubular architecture.
4.Preliminary evaluation of the relationship between the level of plasma of miR-125b-5p and the lesion degree of coronary arteryin in coronary atherosclerotic heart disease
Hui CONG ; Haidan CHU ; Rongrong JING ; Shaoqing JU ; Bingying ZHU ; Xianjuan SHEN ; Xudongn WANG ; Shang WU
Chinese Journal of Laboratory Medicine 2017;40(5):396-399
Objective To investigate the clinical value of circulating miR-125b-5p in coronary atherosclerotic heart disease.Methods With case-control study,80 cases of coronary atherosclerotic heart disease were recruited in Affiliated Hospital of Nantong University from February 2014 to august 2015.According to coronary angiography result they were divided into two groups: there are coronary artery stenosis group(n=49)and control group(n=31).All patients were also divided into non-ST-segment elevation myocardial infarction and ST-segment elevation myocardial infarction group(n=35),unstable angina group group(n=25),stable angina group(n=20).The level of miR-125b-5p before coronary angiograph was detected.By independent sample t test and variance analysis,the levels of miR-125b-5p were compared between the groups of coronary artery stenosis and the group with no stenosis of the coronary artery,the coronary artery lesions in each group,and between the various types of coronary atherosclerotic heart disease respectively.Results MiR-125b-5p expression level of Coronary artery stenosis group(0.35±0.10)was lower than that in group coronary artery with no stenosis(0.95±0.12),the difference was statistically significant(t=24.179,P<0.000 1).With the increase in the number of diseased coronary arteries,miR-125b-5p expression level decreased gradually.There is also statistical significance(t=8.399,P<0.000 1; t=13.067,P<0.000 1)in miR-125b-5p expression among NSTEMI+STEMI,UA and SAP groups.miR-125b-5p expression level was negatively correlated with Gensini score(R2=0.822,P<0.05).The area under the ROC curve(AUC)of miR-125b-5p was 0.86(95%CI 0.67-0.90),and 0.66 was the optimal cut-off value with sensitivity of 81.22%and specificity of 78.62%.Conclusions With the increase of the number of stenosis,plasma miR-125b-5p expression level decreased gradually.The expression level of miR-125b-5p was negatively correlated with the Gensini score of coronary artery,which indicated that the expression level of miR-125b-5p may be a potential biomarker that can reflect the lesion degree of coronary artery.
5.Effect of Compound Yibazhen Granules on Early Renal Injury in Spontaneous Hypertensive Rats
Xinli LIN ; Zhuyuan FANG ; Xuanxuan ZHU ; Haidan WANG ; Rui FU ; Di ZHANG ; Xuefei ZHANG
Herald of Medicine 2016;(2):125-128,129
Objective To observe the antihypertensive effect of compound Yibazhen granules on spontaneous hypertensive rats ( SHR) . Methods Wistar rats were served as normal control group. Sixty SHR were randomly divided into model control group,captopril group,Jane chrysanthemum antihypertension tablet group and compound high dose group,middle dose group and low dose group ( n = 10 each group ) by digital table method. Captopril group was given captopril 30 mg.kg-1 .d-1 ,and Jane chrysanthemum antihypertension tablet group was treated with Jane chrysanthemum antihypertension tablet ( 0. 6 tablet per kg ) , compound Yibazhen granules high dose group, middle dose group and low dose group received compound of 13.18,6.59 and 3.3 mg.kg-1 .d-1 ,respectively. Normal control group and model control group were intragastrically administered with 0.9% sodium chloride solution for 8 weeks. Changes of systolic and diastolic blood pressure of rats and blood urea,creatinine,nitric oxide (NO),nitric oxide synthase (NOS) and angiotensin Ⅱ (Ang Ⅱ) were observed. Results Diastolic pressure of rats in compound Yibazhen granules high dose group, middle dose group and low dose group decreased significantly in 2 weeks. Systolic blood pressure and diastolic blood pressure of compound Yibazhen granules high dose group decreased significantly in 4 weeks,compared with the model control group (P<0.05). Compared with the model control group, concentration of urea and crea in compound Yibazhen granules high dose group, middle dose group and low dose group were significantly lower( P<0.05) . The content of NOS and AngⅡ in rats of compound Yibazhen granules high dose group decreased significantly and the contents of NO increased, which were compared with the model control group ( P<0. 05, P<0. 01 ) . Conclusion The protective effect of compound Yibazhen granules in treating early renal damage in SHR is related to decreasing diastolic blood pressure,concentration of urea,crea and AngⅡ and regulating the levels of NOS and NO.
6.Practice and reflection on teaching of rehabilitation medicine in general practitioners' training
Kaiting LI ; Dawu WANG ; Yuhan KONG ; Juanwen HE ; Jia HE ; Haidan LIN ; Xiaohong LI ; Dingqun BAI
Chinese Journal of Medical Education Research 2016;15(7):742-744,745
The practice of general practice emphasizes that the general practitioner is the backbone, and the primary health institution is the main body, which solves the main health problems of the commu-nity and meets the basic health service needs of the residents. Rehabilitation medicine occupies a prominent position in the community health service, and its related knowledge and skills are the important part of the whole medical education. However, rehabilitation medicine has more teaching contents, less training time and out-of-date teaching model, which have made the effect of rehabilitation medicine teaching not good. In the rehabilitation medicine teaching for training general practitioners, by strengthening the concept of reha-bilitation, elaborating teaching content, optimizing teaching ward-round, and outstanding ability as well as reforming evaluation way, we try to exercise the students' clinical thinking and improve clinical skills, to cultivate outstanding qualified general medical practitioners for community rehabilitation ultimately.
7.Long term follow up of patients with the ileal orthotopic neobladder
Xuede QIU ; Hongyi XU ; Yongfu SHI ; Zehui LI ; Jiongming LI ; Jiansong WANG ; Zhipeng LI ; Kewei FANG ; Guihua CAO ; Haidan LI ; Jin HE ; Shuchen HE
Chinese Journal of Urology 2009;30(10):677-680
Objective To discuss the long term clinical effect of ileal orthotropic neobladder.Methods From 1991 to 1998,79 patients,mean age 55(41~75)years,male 74,female 6,were followed up.The serum creatinine and urea,electrolytes,blood routine,B ultrasonic scan of the neobladder residual urine and IVU or MRU of the patients were followed up.The max transverse diameter of renal pelvis and the max verticaI/level diameter of neobladder were measured in 5,10 to 14,15 years of postoperative when IVU or MRU.All results of different time were compared by the multiple comparisons.The local or distant cancer recurrence and the complications of the operation Were evaluated. Results Sixty-four cases,58 male,6 famle,were long term followed up:mean time was 167 (range,121~216)months.Seven cases died of other diseases.Seven cases had pelvic recarrence.Two cases had urethral recurrence.Three cases died of tumor metastasis.One case had ureter recurrence.Forty-eight patients were alive more than 10 years.The value of the serum creatinine,urea,electrolytes and bloods routine of the patients were normal after 5,10 to 14 and 15 vears postoperative (P>0.05).The max transverse diameter of the renal pelvis in 5,10 to 14 and 15 years Dostoperative were 14.0 mm,14.1 mm and 13.7 mm,respectively,P>0.05.The max vertical/level diameter of the neobladder in 5,10 to 14,15 years of postoperative were 110.4 mm/90.4 mm,111.5 mm/95.3mm and 127.0 mm/97.0 mm,respectively,P>0.05.The residual urine of 5 cases was more than 50 ml and had not increased during follow up.Eight cases with neobladder stone were cured by the intracavitary lithothrypsis.Two cases with uretheral stricture were cured by the intracavitary therapy.Twelve cases of 14 cases with inguinal hernia were cured by reoperation,2 cases accepted conservative treatment.Only 17 cases had no complication involve of the cancer and the operation. Conclusion The upper urinary tract and neobladder of the ileal orthotopic neobladder could be stable for long time,the cure rate of tumor is satisfactory and the lifetime follow up is necessary.
8.Practice and discussion on evaluation system of exploratory experiment's teaching
Haidan LUO ; Yunling XIE ; Donghui WANG ; Hua ZHOU ; Kaiyu TANG ; Xuelan WANG ; Huiling YANG
Chinese Journal of Medical Education Research 2018;17(4):325-329
Exploratory experiment is the key to the experimental physiology,which is universally applicable training of innovative talents.More importantly,it is an important way to cultivate innovative talents of medicine.On the basis of the original multi-subject evaluation system,we have formed a comprehensive evaluation system through refining the scoring item for teachers and students,enriching the forms of student evaluation,and building the student self-assessment.This comprehensive evaluation system is set to promote the development of experimental physiology,especially the exploratory experiments.The practice of this evaluation system effectively improve the objectivity of the evaluation,provide a basis for reform of exploratory experiment,and then promote the training of innovative talents.
9.Perceived quality of patient-centered practice in surgical services designed for elderly orthopedic patients
Fei WANG ; Haifei XIANG ; Weiping WANG ; Haidan SHI ; Liping JIA
Chinese Journal of Geriatrics 2021;40(6):766-769
Objective:To investigate the effects on perceived quality of a management assessment system for orthopedic surgery in elderly patients.Methods:Based on the concept of lean management and process nodes in systems, an information management system was established to evaluate the perceived quality of surgical service for elderly orthopedic patients.Four hundred and two elderly orthopedic patients receiving orthopedic surgery after the implementation of the management system were included in the observation group, while 402 elderly patients undergoing orthopedic surgery before the implementation of management system formed the control group.Results:The incidence of hypothermia in patients admitted to the resuscitation room after surgery was lower in the observation group than in the control group(1.7% vs.4.5%, χ2=5.136、 P=0.023). The incidence of shivering during resuscitation was lower and the satisfaction rate of surgical services was higher in the observation group than in the control group(0.7% vs.3.2%) χ2=5.279、 P=0.022, 96.55(95.58, 97.28)分, vs.87.9(83.2, 89.95)分, Z=-4.16, P=0.00). Conclusions:A perceived quality assessment system composed of lean management and information technology for surgical services can reduce complications and improve satisfaction for patients receiving surgery.
10.Clinical diagnostic value of circulating tumor cells and circulating cell-free DNA combined detection in peripheral blood for breast cancer
Xiaofen ZHANG ; Chunping JIA ; Hongmei CHEN ; Yingjuan SHI ; Shaoqing JU ; Xudong WANG ; Haidan CHU ; Hui CONG
Chinese Journal of Laboratory Medicine 2019;42(8):662-668
Objective To investigate the clinical diagnostic value of circulating tumor cells (CTCs)and circulating cell-free DNA (cfDNA) in peripheral blood samples in breast cancer. Methods From July 2017 to April 2018, 47 patients with BMC (7 in stage Ⅱ, 19 in stage Ⅲ and 21 in stage Ⅳ), 24 patients with benign breast diseases and 28 healthy people were selected. After collecting peripheral blood samples, serum and blood cells were separated. The size-based high-throughput microfluidic chip was used to capture CTCs. The real-time fluorescent quantitative PCR based on Alu sequence was used to detect the length of cfDNA(247 bp, 115 bp)in the serum, and the ratio of amplified products of long and short fragments was used as the index of DNA integrity. The Mann-Whitney U test or Kruskal-Wallis H test was used to compare the differences between the groups and analyze the relationship between CTCs and cfDNA and clinical parameters of breast cancer. The ROC curve was drawn and the area under the curve (AUC) was used to evaluate the feasibility of blood cell CTCs and plasma cfDNA detection as diagnostic criteria. Results The CTCs and cfDNA of 47 BMC patients were analyzed. The CTCs and cfDNA integrity index (Alu 247/115) of BMC patients were significantly higher than those of physical examination patients[(13.98± 12.36)cells / ml vs (1.14 ± 1.35) cells / ml; 0.7687 ± 0.3868 vs 0.5094 ± 0.2456], and the difference was statistically significant(the U value was 126.5,359.0;P<0.001), the area under ROC curve of CTCs was 0.885 (95%CI: 0.805-0.965), cut-off value was 7.68/ml, sensitivity was 80.4%, specificity was 96.4%. The area under ROC curve of Alu 247/115 was 0.727(95%CI: 0.608-0.847), cut-off value was 0.431, sensitivity was 71.7%, specificity was 71.4%. The AUC of CTCs and Alu 247/115 was 0.919 (95%CI 0.854-0.984), which was higher than the single test of each indicator. Conclusions CTCs and cfDNA may be the potential biological indicators for breast cancer diagnosis. The combined detection of CTCs and cfDNA maybe improve the diagnosis rate of breast cancer patients.