1.Long non-coding RNA SPRY4-IT1 expression in esophageal squamous cell carcinoma and its effects on cell growth
Haiwei XIE ; Fangjun CHEN ; Bin ZHU ; Gang CAO ; Lei JIN ; Guozhi ZHOU ; Jin LV ; Xiufeng CAO
Chinese Journal of Clinical Oncology 2013;(17):1011-1015
Objective:This study aimed to clarify the correlation of SPRY4-IT1 expression with the clinicopathological character-istics and prognosis of patients with esophageal squamous cell carcinoma (ESCC), as well as the role of SPRY4-IT1 in promoting ES-CC cell growth. Methods:Quantitative real-time polymerase chain reaction for SPRY4-IT1 expression was performed on 50 paired can-cerous and adjacent non-cancerous esophageal specimens. Small interfering RNA was used to suppress SPRY4-IT1 expression to fur-ther explore its role in tumor progression. Cell viability was tested in vitro by MTT assay (OD=490 nm), and cell apoptosis and cell cy-cle were investigated by flow cytometry. Results:We found markedly elevated SPRY4-IT1 expression in cancerous tissues compared with adjacent non-cancerous tissues (90%, P<0.01). Relative SPRY4-IT1 expression levels were correlated with some clinicopathologi-cal characteristics, such as tumor size (χ2=5.333, P=0.021), elevated TNM (2009) stage classi fi cation (χ2=5.556, P=0.018), and de-creased overall survival rates (χ2=5.296, P=0.021). SPRY4-IT1 expression level was not correlated with patient age, gender, smoking status, or alcohol consumption (all P>0.05). Further experiments showed that SPRY4-IT1 expression levels were significantly higher in three ESCC cell lines than in the normal human esophageal epithelial cell line Het-1A. In vitro assays of the ESCC cell line KYSE30 demonstrated that knockdown of SPRY4-IT1 expression by small interfering RNA reduced cell growth, mediated cell cycle arrest at the G0-G1 phase, and promoted cell apoptosis (all P<0.01). Conclusion:SPRY4-IT1 was overexpressed in ESCC tissues and ESCC cell lines and promoted the growth of ESCC cells. The dysregulated expression of long non-coding RNA SPRY4-IT1 may play an important role in the process of ESCC development and may be developed as a useful biomarker for the diagnosis and prognosis of ESCC.
2.Affection of Zidan decoction containing serum on endometrial receptivity in uterine immunity environment
Qian CHEN ; Zhe JIN ; Qiudan LU ; Ying CAO ; Nan LI
International Journal of Traditional Chinese Medicine 2012;34(3):220-223
Objective To investigate the effect of Zidan decoction containing serum on endometrial receptivity in uterine immunity environment,and then to discuss if it's effect on endometrial receptivity through its controlling the expression of cell factors which related to endometrial receptivity.Methods Mice's endometrium tissue samples and uterine Natural Killer (UNK) cells were obtained from pregnant mice on gd 4.5 days and gd 12.5 days.After endometrial glandular epithelial cells,endometrial stromal cells and UNK cells were separated through primary culture,epithelial cells and endometrial stromal cells at rate of 1 ∶ 1 a certain concentration UNK were mixed together and highly purified-cultured.Different concentrations of Zidan decoction was acted on the culture solution.FBS,black and aspirin control group were setup for contrast.The expression of LIF and VEGF which influenced by Zidan decoction in the immunity environment was assayed by Realtime-PCR.Results The expression of LIF and VEGF was significantly increased in the immunity environment of uterine (in 10% Zidan decoction group,10% aspirin group and 10% blank control group:LIF was 2.10±0.20,1.98±0.14,0.90±0.05 separately,and VEGF was 3.17±0.31,2.52±0.09,0.92±0.06respectively,P<0.05).Conclusion Zidan decoction could improve endometrial receptivity by increasing the expression of cell factor which related to endometrial receptivity with the help ofUNK paracrine.
3.Assessment of Bax gene transfer influence on survival of heterotopic cardiac allograft in rats by high-frequency echocardiography
Jiamei JIN ; Yuhui ZHANG ; Ming CHEN ; Ye ZHU ; Hao CAO
Chinese Journal of Ultrasonography 2013;22(11):986-990
Objective To evaluate the value of high-frequency echocardiography in assessing Bax gene transfer influence on survival of heterotopic cardiac allograft in rats.Methods Thirty rat models of heterotopic heart transplantation were established.Group A received heart transplantation only; Group B received cyclosporin (CsA) after operatiom Group C received ultrasound targeted microbubble destruction (UTMD) combined with Bax-shRNA.All rats were tested by high-frequency echocardiography at day 1,3,6 after transplantation.The ultrasound parameters included left ventricular internal dimension diastole (LVIDd),left ventricular internal dimension systole(LVIDs),left ventricular ejection fraction(LVEF),left ventricular thickness (LVT),left ventricular thickening (LVTR) and so on.The pathologic examinations were carried out on five rats at day 6 after echocardiography exam.The other rats in each group were observed for the survival time of cardiac allograft.Results ①The left ventricular long axis view and the apical four chamber view could be displayed clearly by high-frequency echocardiography.②The survival time of allografts in group C was (16.21 ± 5.01)d,which was longer than that in group B [(11.14 ± 1.72)d,P < 0.05] and group A [(7.26 ± 1.50)d,P <0.01].③LVT index got higher after transfection.At day 6,LVEF of group A was lower than group B and C markedly(P <0.05) while no significant difference was found between group B and C(P >0.05).At day 3 and 6,LVT of group A and B were higher than group C (P <0.05) while LVTR was lower(P <0.05).Conclusions The Bax gene transfer influence on survival of heterotopic cardiac allograft in rats could be evaluated accurately by high-frequency echocardiography which could assess cardiac structure and function.LVT and LVTR can be considered as early evaluation indexes for its high sensitivity over LVEF.
4.Effect of Acupuncture at Shixuan (EX-UE11) plus Rehabilitation on Upper-limb Spasticity After Cerebral Stroke
Liping CHEN ; Haiping BIAN ; Jin WANG ; Defeng CAO
Shanghai Journal of Acupuncture and Moxibustion 2016;35(10):1154-1157
Objective To observe the clinical efficacy of acupuncture at Shixuan (EX-UE11) plus rehabilitation in improving the upper-limb spasticity and hand function after cerebral stroke. Method Sixty patients with upper-limb spasm due to cerebral stroke were randomized into two groups. The observation group was intervened by conventional acupuncture plus needling Shixuan and rehabilitation training, while the control group was by conventional acupuncture plus rehabilitation training. The Modified Ashworth Scale (MAS) and Modified Barthel Index (MBI) were evaluated before and after the intervention. The effects of the two methods in improving the upper-limb spastic intensity, hand function, and activities of daily living after cerebral stroke were evaluated based on the scores before and after the intervention. Result After treatment, the upper-limb spastic intensity and muscle tension decreased in both groups, and there was a significant difference in comparing the MAS score between the two groups (P<0.05); the improvement of MBI in the treatment group was more significant than that in the control group (P<0.05). Conclusion Acupuncture at Shixuan plus rehabilitation can produce a more significant efficacy in treating upper-limb spasm, improving hand function and activities of daily living after cerebral stroke, compared to the conventional acupuncture plus rehabilitation.
5.Detection of interleukin-10 in sera and culture supernatants of CD4 +CD25 + T cells from children with atopic dermatitis
Wanwan JIN ; Boying HU ; Xinhua MA ; Shisheng CHEN ; Yu CAO
Chinese Journal of Dermatology 2013;(1):13-15
Objective To determine the level of interleukin (IL)-10 in sera and culture supernatants of CD4+CD25+T cells from children with atopic dermatitis (AD),and to evaluate its relationship with clinical course and severity of AD.Methods Totally,46 children with AD and 31 healthy controls were included in the study.All the patients were divided into 3 groups,i.e.,mild (n =10),moderate (n =16) and severe (n =20) group,according to severity scoring of atopic dermatitis (SCORAD) score.Venous blood samples were obtained from the patients and healthy controls.CD4+CD25+ regulatory T cells were separated from the blood samples by magnetic cell sorting (MACS) system in two steps and cultured in vitro.Enzyme linked immunosorbent assay (ELISA) was conducted to quantify the level of IL-10 in sera and culture supernatants of CD4+CD25 + T cells from these subjects.Analysis of variance was carried out to compare the level of supematant and serum IL-10 between the patients and controls,and Pearson correlation analysis to assess the relationship between the level of IL-10 and SCORAD score.Results The patients with mild,moderate and severe AD showed a similar serum IL-10 level compared with the healthy controls ((43.10 ± 25.07) pg/ml,(68.40 ± 36.65) pg/ml and (55.55 ± 41.97) pg/ml vs.(58.27 ± 36.84) pg/ml,all P > 0.05).The level of supernatant IL-10 secreted by CD4+CD25+ T cells from the controls was significantly higher than that from the patients with severe AD ((55.15 ± 11.15) pg/ml vs.(27.25 ± 7.01) pg/ml,P < 0.05),but similar to that from the patients with mild and moderate AD ((52.96 ± 11.69) pg/ml and (49.86 ± 9.18) pg/ml,respectively,both P > 0.05).The level of secreted IL-10 was negatively correlated with SCORAD score (r =-0.757,P < 0.01),whereas the serum level of IL-10 showed no statistical correlation with SCORAD score.Conclusion CD4+CD25+ T cells and IL-10 may be implicated in the development of AD.
6.Correlation between immune response to hepatitis B vaccine and HLA DRB1*07 DRB1*04 DRB1*1001.DQB1*0401 genes in Han population in Ningbo
Jin-Feng ZHAO ; Hai-Tang CHEN ; Guang-Wen CAO ;
Academic Journal of Second Military Medical University 2000;0(08):-
Objective:To study the correlation between responses to Hepatitis B and HLA DRB1*07 DRB1*04 DRB1*1001. DQB1*0401 genes in Han population in Ningbo.Methods:A total of 240 Han people living in Ningbo received the routine vaccination of recombinant hepatitis B vaccine.The serum levels of anti-HBs antibody were examined and the subjects were divided into 2 groups according to tile results:negative responses group(n=120.anti-HBs D
7.Effects of spinal pedicle screw internal fixation at different extraversion angles Biomechanical evaluation
Qun CHEN ; Zhengshuai JIN ; Xiaojian CAO ; Quan JIANG
Chinese Journal of Tissue Engineering Research 2008;12(13):2573-2575
BACKGROUND: It has been demonstrated that pedicle screw internal fixation influencing factors involve screw structural morphology, thread parameter, bone density, moment size for tightening screw during operation, and depth of screw placement. There is little known about the correlation of pullout strength of spinal pedicle screw with device for transverse traction to extraversion angle. OBJECTIVE: To assess effect of the spinal pedicle screw with device for transverse traction on pullout strength at different extraversion angles. DESIGN: Repeated measurement. SETTING: Center for Bone Joint, the First Affiliated Hospital of Nanjing Medical University. PARTICIPANTS: This study was performed at Laboratory for Material Mechanics, Hehai University between June and November 2003. A total of 18 adult dried lumbar vertebrae (L1-5) were provided by Department of Anatomy, Nanjing Medical University, and recruited for this study. The protocol was approved by the hospital's Ethics Committee. The pedicle screw was made of stainless steel. Each pedicle screw had a diameter of 5.5 nun, total length of 150 nun (thread part 50 into included), and the same thread parameter. Electrical universal material machine (EW type) was provided by Laboratory for Material Mechanics of Hehai University. METHODS: Bone density was measured with a single photon bone density determinator. According to the bone density, the lumbar vertebrae were numbered and randomly divided into 3 groups with 6 lumbar vertebrae in each: extraversion angle 5 ° group, extraversion angle 15° group, and extraversion angle 30° group. ① Installation of pedicle screw and clamping apparatus: According to Wein-Stein method, one entry-point was selected at each side of lumbar vertebra, and at the sametime, extraversion angle 5°, 15° ,and 30° were respectively defined for extraversion angle 5° , 15° ,and 30° groups. A 50 mm-depth pinhole was drilled with a drill bit with a diameter of 3.0 nun. Pedicle screw was screwed into 50 nun, and its end part was connected to the device for transverse traction. Spinal vertebrae and the device for transverse traction were fixed with a specially made clamping apparatus. ② Determination of pullout strength and observation of pedicle and vertebral injury: Spinal vertebrae, on which pedicle screw and device for transverse traction were installed, was placed on a EW electrical universal material machine together with clamping apparatus for determining the pullout strength of pedicle screw. Sensor was connected to a computer to draw strength-displacement curve. The wave crest of the curve was considered the maximum pullout strength. At the same time, injuries to pedicle and vertebra caused by pullout of pedicle screw were observed. MAIN OUTCOME MEASURES: Pullout strength and injuries to pedicle and vertebra. RESULTS:①The mean maximum pullout strength of pedicle screw was respectively 0.878 167, 1.420 333, and 2.154 167 KN for extraversion angle 5° , 15 ° , and 30° groups. There was significant difference among the 3 groups (F = 12.554 22, P < 0.01). ② In the extraversion angle 5° group, 4 patients presented with cortical bone fracture which occurred at the entrance for pedicle screw, and 2 patients presented with fragmentation of junctional zone between pedicle and vertebral posterior edge; In the extraversion angle 15° group, 1 patient presented with cortical bone fracture which occurred at the entrance for pedicle screw, 4 patients presented with fragmentation of junctional zone between pedicle and vertebral posterior edge, and 1 patient presented with vertebral posterior coronal fragmentation; In the extraversion angle 30° group, 1 patient presented with cortical bone fracture which occurred at the entrance for pedicle screw, 2 patients presented with fragmentation of junctional zone between pedicle and vertebral posterior edge, and 3 patients presented with vertebral posterior coronal fragmentation. There was statistical significance in the intergroup rank-sum test (P < 0.01).CONCLUSION: During application of pedicle screw with device for transverse traction, proper accrescence of extraversion angle can increase pullout strength of the screw and enhance fixative strength, and excessive extraversion angle easily injures vertebra.
8.Lipopolysaccharide Down-regulates SR-BI Expression in a Nucleus Factor-κB Pathway-dependent Manner
Dongli CAO ; Lijin WANG ; Tao JIN ; Hui CHEN
Chinese Journal of Primary Medicine and Pharmacy 2011;18(8):1011-1013
Objective To investigate the changes of cholesterol efflux,the scavenger receptor class B type Ⅰ(SR-BI) protein expression in THP-1 maerophage derived foam cells treated with Lippolysaecharide (LPS), and to discover the role of NF-κB pathway in this process.Methods The foam cells were treated with LPS along or treated with N-p-Tosyl-L-phenylalanine chloromethyl ketone(TPCK) for 24 h.The protein levels of SR-BI and intranuclear NF-κB p65 were measured by Western blotting.Cellular lipid accumulation was determined by high performance liquid ehromatograpby analysis.Cholesterol efflux was determined by FJ-2107P type liquid scintillator.Results The expression of SR-BI was decreased after treated with LPS,while the intranuclear NF-κB p65 protein level was increased by LPS.The results also showed that cellular lipid accumulation was increased ,while the cellular cholesterol efflux was decreased in THP-1 maerophage derived foam cells after exposed to LPS for 24 h and these changes can be reversed partly by pretreatment with TPCK.Conclusion LPS could down-regulate the expression of SR-BI, promote the accumulation of lipid and decrease cellular cholesterol efflux in THP-1 maerophage derived foam cells ,which should be related to the TLR4/NF-κB dependent pathway.
9.Influence of Gastroesophageal Reflux Disease Morbidity Rate by High Salt Diet in Mare Island Area
Wuheng TANG ; Hai JIN ; Songcai LI ; Chunyu CAO ; Guoguang CHEN
Journal of Medical Research 2006;0(11):-
12,9.41%(148/1578);RDQ≤12,91.58%(1430/1578).Logistic multiple regression analysis of gastroesophageal reflux correlation factor studied:OR= 2.781.Conclusions The results showed:there were close correlation of high salt diet and GERED.
10.Application of flow cytometry in the diagnosis of X-linked hyper-IgM syndrome
Ruiming CAO ; Tongxin CHEN ; Xi WANG ; Yazhong ZHU ; Yingying JIN
Chinese Journal of Laboratory Medicine 2008;31(5):509-512
Objective To develop a flow cytometrie assay to for diagnosis of X-Linked Hyper.1gM Syndrome(XHIM).Methods Heparinized peripheral blood obtained from patient,mother and a healthy control was diluted with RPMI- 1640(unstimulated control)or with RPMI-1640 containing 15μl PMA(1 rig/μl)and 6 trl ionomycin(50 ng/μl)(stimulated cell).Using directly labeled antibodies,we have examined CD40 ligand levels on CD3+ CD8- lymphocyte surface,and CD69 levels on CD;lymphocyte Surface to determine whether the cells were activated.Results CD69 levels on CD3+ lymphocyte surface from stimulated group and from unstimulated group were above 96% and below 3%,respectively.CD40L levels Oil CD3 CDs-T lymphocyte surface from stimulated group were 0.8% (patient),60.04%(mother) and 62.87%(healthy contr01).CD40L levels on CD3+ CD8-T lymphocyte surface from unstimulated group were 0.88% (patient),4.15%(mother)and 5.51%(healthy contr01).Conclusion This flow cytometric assayis accurate and convenient,which Can be used in neonatal screening.