1.Effects of RhoGDI2 gene silencing by siRNA on proliferation and invasion of colon carcinoma cell lines
Wenrui WANG ; Hongli YAN ; Lijun MA ; Siqi LIU ; Zhenjing JIN
Chinese Journal of Immunology 2017;33(4):549-553
Objective:To study the silencing gene expression level of RhoGDI2 small interfering RNA(siRNA)and the colorectal cancer cell malignant behaviors such as cell proliferation,migration,invasion.Methods:Testing RhoGDI2 expression using Westen blot analysis and Real-time reverse transcription polymerase chain reaction(RT-qPCR)in the colorectal cancer cell lines of RKO,HT29,SW620,SW480,HCT116.The siRNA of RhoGDI2 with Lipofecta mineTM2000 was transfected into target cells,as well as negative control and normal control groups.Cell counting kits(CCK-8)to detect proliferation,Wound healing assay and the Transwell plate migration and invasion was detected.The epithelial-mesenchymal transition(EMT)relevant protein E-cadherin/Vimentin expression was detected.Results:Human colon cancer cell lines RhoGDI2 expression levels decreased in the order of RKO,HT29,SW620,SW480,HCT116:siRNA inhibited RhoGDI2 expression rate of RKO cell by 70%;in silence group,negative control group and blank contro1 group,the proliferation rates were(0.683±0.013),(0.866±0.088),(0.905±0.008),P<0.05;Wound healing assay and Transwell assay suggested RhoGDI2 silencing could inhibit migration;siRNA interference of colon cancer cells downregulated Vimentin,but upregulated E-Cadherin protein.Conclusion:RhoGDI2 down-regulation could significantly inhibit the cell proliferation,migration,invasion of colon cancer cell.
2.The association between peripheral blood circulating follicular helper T cells and antibody-secreting B cells in patients with ankylosing spondylitis
Siqi LONG ; Li MA ; Hong LI ; Qingqing WU
Chinese Journal of Rheumatology 2016;20(3):164-168
Objective To investigate the relationship between peripheral blood circulating follicular helper T cells and antibody-secreting B cells in patients with ankylosing spondylitis (AS). The role of circulating follicular helper T cells and antibody-secreting B cells in the pathogenesis of AS were explored. Methods Flow cytometry was used to detect the levels of peripheral blood CD4 +CXCR5+ T (cTfh), CD4+CXCR5+PD-1+ T, CD4+CXCR5+ICOS+ T, CD19+ B, CD19+CD38+ B cells in 59 patients with AS (including 36 cases of active AS patients and 23 cases of inactive AS patients). In addition, twenty healthy persons were selected as the controls. Data analysis were performed by independent-sample t test, One way ANOVA analysis, Pearson's and Spearman's correlation test. Results The percentages of cTfh [(26.8 ±10.4)%], CD4+CXCR5+PD-1+T [(12.1±14.0)%], CD4+CXCR5+ICOS+T [(13.6±9.5)%], CD19+CD38+B [(80.7±13.0)%] in the peripheral blood of AS group were significantly higher than healthy controls [(15.6 ±4.5)%, (6.4 ±2.4)%, (9.4 ± 4.5)%, (68.2±13.0)%] (t=6.663, P<0.01; t=2.999, P<0.01; t=2.573, P<0.05; t=2.712, P<0.01). The percentages of cTfh in the active AS group [(30.2 ±11.0)%] were significantly higher than those in the inactive AS group [(21.4±6.5)%] and HC (t=3.444, P<0.01;t=7.004, P<0.01). The percentage of CD19+CD38+B[(85.1±10.0)%] in the peripheral blood of active AS group was significantly higher than that of the inactive AS group [(73.8 ±14.2)%] and HC (t=3.561, P<0.01;t=5.410, P<0.01). The relationship between Bath AS disease activity index (BASDAI) and the percentage of cTfh, CD19+CD38+B was a positive correlation (r=0.442, P<0.01; r=0.405, P=0.001), and significant positive correlation was observed between the percentages of cTfh and CD19+CD38+B cells (r=0.420, P=0.001). Conclusion CD4+CXCR5+cTfh cells are significantly increased in peripheral blood in AS patients with aberrant CD19+CD38+ antibody-secreting B cells, suggesting that cTfh and CD19+CD38+antibody-secreting B cells may play an important role in the pathogenesis of AS .
3.Analysis difference of lymphocyte subgroups between children with severe pneumonia and non-severe pneumonia
Siqi LONG ; Li MA ; Min CHEN ; Hong LI
Chinese Journal of Immunology 2016;32(3):401-404
Objective:To observe the levels of lymphocyte subgroups in children with severe pneumonia and non -severe pneumonia , and to investigate the change of celluer immune funciton of patients , moreover provide available reference for treatment.Methods:Flow cytometry instrument was used to detect peripheral blood CD 3+T, CD3+CD4+T, CD3+CD8+T, CD19+B lymphocytes and NK cells in 27 children with severe pneumonia and 28 children with non-severe pneumonia.In addition 20 heathy children were selected as the controls .Results:Compared with the heathy control group ,the levels of CD3+T,CD3+CD4+T,CD3+CD8+T lymphocytes and NK cells in children with severe pneumonia decreased significantly (P<0.01,P<0.05,P<0.01,P<0.05),while CD4/CD8 ratio,CD19+B cells were significantly increased (P<0.05,P<0.01).The level of CD3+CD8+T lymphocytes in children with non-severe pneumonia significantly lower than the heathy control group (P<0.01),but the level of CD19+B cells in children with non-severe pneumonia significantly higher than the heathy control group (P<0.01).Compared with non-severe pneumonia group.The levels of CD3+and CD3+CD8+T lymphocytes in children with severe pneumonia decreased significantly (P<0.01),however CD19+B cells were significantly increased ( P<0.01 ) .Conclusion: Immune dysfunction exists in both of children with severe pneumonia and non-severe pneumonia.Children with severe pneumonia has more serious damage in inmmunity .This provides certain theoretical basis for clinical e-valuation and treatment .
4.Reliability and Validity of Job Satisfaction Scale for Public Health Practitioners
Siqi WANG ; Yansong SONG ; Shuhua XI ; Yanan MA
Journal of China Medical University 2017;46(6):536-539
Objective The study aimed to explore the reliability and validity of public health practitioners 'job satisfaction scale. Methods A questionnaire survey was conducted with professional and technical personnel in the province engaged in the work of public health of investigation;they were enrolled through the multi?stage cluster random sampling method for establishing the scale 's internal consistency reliability and structur?al validity. Results The Cronbach's alpha coefficient of the total satisfaction scale was 0.900. The Cronbach's alpha coefficients were 0.896, 0.781,and 0.799 for career development and achievement,relationship and career retention,and income and promotion latitudes,respectively. Four principal components were extracted and the results showed that the cumulative contribution rate was 69.72%and the factor analysis results were basically consistent with the theoretical structure. Conclusion The scale is thus considered to be a reliable and effective measurement tool assessing public health practitioners'job satisfaction,as it had good reliability and validity.
5.RhoGDI2 expression and clinical significance in colorectal cancer
Siqi LIU ; Lijun MA ; Hongli YAN ; Wenrui WANG ; Zhenjing JIN
Chinese Journal of Immunology 2017;33(1):108-111
Objective:To explore the expression and clinical significance of RhoGDI2 in colorectal cancer. Methods:Immuno-histochemistry was used to identify RhoGDI2 expression in clinical samples of colorectal cancer tissues,para-tumorous tissues and lymph node metastasis tissues. The relationships between CRC clinical factors and survival were analyzed. Results: RhoGDI2 expression contributed positively with tumor progression and metastasis in clinical tissues. It was associated with the stage of the tumor,lymph node metastasis, remote metastasis, venous invasion and vessel invasion. Patients with higher RhoGDI2 expression had poorer overall survival. Conclusion:RhoGDI2 showed high expression in colorectal cancer and it was associated with the stage of the tumor,lymph node metastasis,remote metastasis, venous invasion and vessel invasion. Patients with higher RhoGDI2 expression had poorer overall survival.
6.Infrared spectrum analysis of Taichong(LR3)and Taixi(KI3)points before and after menstruation in healthy women
Yingbo HAO ; Siqi MA ; Xueyong SHEN ; Jianzi WEI ; Yanping YANG
Journal of Acupuncture and Tuina Science 2024;22(2):160-166
Objective:To investigate the variations in infrared radiation at Taichong(LR3),Taixi(KI3),and control points before and after menstruation and to examine the infrared radiation patterns associated with Yuan-primordial points of Zang-Fu organs during the physiological menstrual cycle. Methods:Using a point infrared radiation spectrum detection system,we detected the infrared radiation spectra of Taichong(LR3),Taixi(KI3),and the control points located 1 cm away from the two points,in a range of 1.50-18.00 μm wavelengths during the premenstrual,menstrual,and postmenstrual phases in 32 healthy adult women.Subsequently,data mining and analysis were conducted. Results:Before,during,and after menstruation,the infrared spectral shapes of bilateral Taichong(LR3),Taixi(KI3),and their control points were generally consistent,with characteristic infrared spectral wavelengths located at 11.25 μm.Prior to menstruation,the total intensity of infrared radiation at the right Taixi(KI3)was significantly lower than that at the control point(P<0.05),and that at the left Taichong(LR3)was significantly lower than that at the control point(P<0.01).During and after menstruation,the total infrared radiation intensity at both Taixi(KI3)was significantly lower than that of the control point(P<0.05).The wavelength points exhibiting significant differences in the infrared radiation intensity between points and control points were concentrated at the primary peak of 7.50-14.25 μm and the secondary peak of 15.00-17.25 μm. Conclusion:During different menstrual phases,the infrared radiation spectra of Taichong(LR3)and Taixi(KI3)exhibited distinct point specificity,mainly evident in the infrared radiation intensity and wavelength.
7.Risk factors for symptomatic intracranial atherosclerotic stenosis: a comparison of anterior circulation and posterior circulation
Yan ZHAO ; Haoyuan MA ; Yaxin DUAN ; Hanlei PEI ; Siqi CHENG ; Guodong XU ; Peiyuan LYU
International Journal of Cerebrovascular Diseases 2021;29(1):13-17
Objective:To investigate the risk factors for anterior circulation and posterior circulation symptomatic intracranial atherosclerotic stenosis (sICAS).Methods:The clinical data of patients admitted to Hebei General Hospital for ischemic stroke or transient ischemic attack (TIA) and diagnosed with sICAS by digital subtraction angiography from May 2019 to May 2020 were retrospectively included. The patients were divided into anterior circulation group and posterior circulation group according to the stenosis sites, and the distribution of sICAS and its risk factors were analyzed.Results:A total of 134 patients with sICAS were enrolled, including 82 males (61.2%) and 52 females (38.8%). Their age was 60.28±11.46 years; 115 (85.8%) had ischemic stroke and 19 (14.2%) had TIA. There were 92 patients (68.7%) in the anterior circulation group and 42 (31.3%) in the posterior circulation group. Body mass index (BMI), systolic and diastolic blood pressure levels, as well as the proportion of patients with hypertension, diabetes, smoking and drinking in the posterior circulation group were significantly higher than those in the anterior circulation group (all P<0.05). Multivariate logistic regression analysis showed that higher BMI (odds ratio [ OR] 1.191, 95% confidence interval [ CI] 1.029-1.379; P=0.019), hypertension ( OR 4.073, 95% CI 1.135-14.616; P=0.031) and diabetes ( OR 2.783, 95% CI 1.149-6.738; P=0.023) were independently correlated with the posterior circulation sICAS. Conclusions:Compared with anterior circulation, high BMI, hypertension and diabetes are the independent risk factors for posterior circulation sICAS.
8.Efficacy of remimazolam-propofol-sufentanil for anesthesia in patients undergoing painless gastroscopy
Xingpeng XIAO ; Ming GUO ; Lan MA ; Siqi LI ; Xuan HE ; Yeda XIAO ; Zhongyuan XIA
Chinese Journal of Anesthesiology 2021;41(2):192-194
Objective:To evaluate the efficacy of remimazolam-propofol-sufentanil for anesthesia in patients undergoing painless gastroscopy.Methods:Eighty American Society of Anesthesiologists physical statusⅠor Ⅱ patients, aged 20-59 yr, weighing 44-69 kg, scheduled for elective painless gastroscopy, were divided into 2 groups ( n=40 each) using a random number table method: remimazolam-propofol-sufentanil group (group RPS) and propofol-sufentanil group (group PS). The patients in group RPS received successive intravenous injection of sufentanil 0.1 μg/kg, remimazolam 0.15 mg/kg and propofol (at a rate of 4 mg/s). The patients in group PS received intravenous injection of sufentanil 0.1 μg/kg and propofol (at a rate of 4 mg/s). When Observer′ s Assessment of Alertness/Sedation Scale score was 0, gastroscopy was performed.The consumption of propofol, time of anesthesia, time for gastroscopy, emergence time and discharge time were recorded.The number of intraoperative assisted respiration cases, body movement and occurrence of adverse reactions at the time of discharge were observed. Results:Compared with group PS, the consumption of propofol was significantly decreased, and the time of anesthesia, emergence time and discharge time were shortened in group RPS ( P<0.05). There was no significant difference in the time for gastroscopy, the number of intraoperative assisted respiration cases, body movement and the occurrence of adverse reactions at discharge time between the 2 groups ( P>0.05). Conclusion:Remimazolam-propofol-sufentanil produces better efficacy for anesthesia than propofol-sufentanil in patients undergoing painless gastroscopy.
9.Effects of IL-12 combined with GM-CSF on apoptosis of H22 hepatoma cells
Siqi LIU ; Qian ZHANG ; Xue SHAO ; Shengnan JIA ; Jingting MA ; Liulan PAN
Clinical Medicine of China 2017;33(7):591-595
Objective To investigate the effects of granulocyte macrophage colony-stimulating factor (GM-CSF) combined with interleukin-12 (IL-12) genes on apoptosis of hepatoma cells.Methods The hepatoma cell lines were cultured in vitro and were divided into four groups: GM-CSF transfection group,IL-12 transfection group,GM-CSF and IL-12 co-transfection group,negative control group (empty load group),respectively.The PIB-CMV3-GM-CSF and PIB-CMV3-IL-12 eukayotic expression vector was built,and 36 h after transfection,fluorescence microscope was used to detect the transfection effect;the expression level of IL-12,GM-CSF,p53,p38 and C-JUN mRNA were detected by RT-PCR,and Western blot was used to examine the expression level of IL-12,GM-CSF,p53,p38 and C-JUN protein.In addition,the flow cytometry was applied to detect cell apoptosis.Results Through fluorescence microscope,green fluorescence was observed in cells of GM-CSF transfection group,IL-12 transfection group,GM-CSF and IL-12 co-transfection group,indicating that the plasmid has successfully transferred into cells.In addition,the expression of p53mRNA in empty load group,GM-CSF transfection group,IL-12 transfection group,GM-CSF and IL-12 co-transfection group were 1.2±0.10,4.3±0.98,4.2±0.34,9.2±0.87,and the protein expression were 1.0±0.10,3.6±0.34,3.8±0.30,5.0±0.60.Compared with the empty load group,the expression level of p53 mRNA and protein were significantly increased in the three plasmid transfection groups (P<0.01).The expression of p53 mRNA and protein were significantly increased in co-transfection group than GM-CSF group and IL-12 group (P<0.01),while in the comparison with GM-CSF transfection group and IL-12 transfection group,the expression level of p53mRNA and protein in the co-transfection group could be improved to a higher degree(P<0.01).Meanwhile,p38 C-JUN mRNA expression levels in empty load group,GM-CSF transfection group,IL-12 transfection group,GM-CSF and IL-12 co-transfection group were as follows: 7.5± 0.9,3.5±0.45,3.7±0.25,1.0±0.11,while p38protein expression levels were 10.1±1.03,6.1± 0.67,7.1 ± 0.61,1.0 ± 0.12,respectively,C-JUN mRNA expression levels were 11.2 ± 1.20,4.1 ± 0.19,3.3 ± 0.30,1.0 ± 0.01,separately,C-JUN protein expression levels were 2.25 ± 0.2,1.8 ± 0.13,1.4 ± 0.12,1.0 ± 0.09.P38, C-JUN mRNA and protein levels were significantly reduced in the three plasmid transfection groups compared with the empty load group (P<0.01).The expression of p38,C-JUN mRNA and protein were reduced to a lower degree in co-transfection group than in GM-CSF transfection group and IL-12 transfection group (P<0.01).Flow cytometer showed that the hepatoma cell apoptosis rate of the empty load group,GM-CSF transfection group,IL-12 transfection group,co-transfection group were (3.43±0.9)%,(5.87±1.02)%,(7.32±1.1)%,(17.47±2.11)%,the rates of the three plasmid transfection groups were significantly higher than that of the empty load group (P<0.01).And the apoptosis rate was significantly increased in the co-transfected group compared with other plasmid groups (P<0.01). Conclusion The combination of GM-CSF and IL-12 could significantly accelerate the apoptosis of hepatoma cells by up-regulating the expression of p53,and down-regulating the expression of p38 and C-JUN.
10.Preparation of polyvinyl alcohol/lota-carrageenan scaffolds and its biocompatibility
Jing CUI ; Yabin ZHANG ; Siqi MA ; Yanjie XIONG ; Man CUI ; Shuofeng LI ; Pengcheng CHE ; Fanglian YAO ; Hong SUN
Chinese Journal of Tissue Engineering Research 2017;21(2):215-220
BACKGROUND:Polyvinyl alcohol (PVA) hydrogel with similar porous structure and mechanical properties to the natural cartilage is very suitable for the repair of articular cartilage. However, the pure PVA hydrogel after lyophilization wil be accompanied by the shrinkage of the polymer network and the col apse of the pores, leading to the inhomogeneous performance of the material even in the state of re-swel ing. Addition of the active polymer wil increase the cel adhesion ability of PVA hydrogel. OBJECTIVE:To construct PVA/lota-carrageenan (l-CA) composite materials with different mass fractions of l-CA and evaluate the biocompatibility with vascular endothelial cel s. METHODS:PVA/l-CA composite films with different contents of l-CA were fabricated and then co-cultured with vascular endothelial cel s. Attachment, proliferation and morphological changes of vascular endothelial cel s on the composite were observed by scanning electron microscope and MTT assay to evaluate its biocompatibility. PVA/l-CA three-dimensional scaffold with different contents of l-CA were constructed, and hemolysis experiment was conducted according to the biological evaluation standards of medical devices, and the porosity and pore size were observed using scanning electron microscope. RESULTS AND CONCLUSION:In vitro experimental results showed that the addition of l-CA could significantly increase the biological activity of PVA hydrogel, and promote the cel attachment and proliferation on the scaffold. The hemolysis rate of each experimental group was less than 5%(the accepted safety standard), suggesting that the composite materials were in accordance with the standard of medical devices for hemolysis experiment. These findings indicate that the composite scaffolds with 20%-30%l-CA possess the pore size suitable for cel growth and proliferation and the porosity beneficial for transportation of nutrients and metabolites, which can serve as an excel ent scaffold for tissue engineering.