1.Optimization of Vacuum Belt Drying Process ofChuan-XiongConcrete by Box-Behnken Design
Ruiting JIN ; Xuefeng LI ; Yonghua TAO ; Yun WU ; Zhenzhong WANG ; Wei XIAO
World Science and Technology-Modernization of Traditional Chinese Medicine 2015;(7):1454-1459
This study was aimed to optimize the belt drying process conditions ofChuan-XiongConcrete. On the basis of the single factor experiment with drying temperature, feeding speed, drying time as three main variables and the transfer rate of ferulic acid as variable, the optimization of the belt drying process conditions ofChuan-XiongConcrete was identified using Box-Behnken Design. The results showed that the optimal drying parameters were as follows. The feeding speed was 1 kg·h-1. The drying temperature was 95℃. The drying time was 90 min. The transfer rate of ferulic acid was 95.48%. Three verification experiments were taken under these technological parameters. And the average transfer rate of ferulic acid was 95.71%, which was close to the predicted value of the model. It was concluded that the belt drying process of Chuan-Xiong Concrete was steady and feasible, which can be used in the drying process ofChuan-XiongConcrete.
2.Application of the rapid influenza testing during 2007-2008 flu season in Beijing city
Ran LI ; Bin CAO ; Shufeng CUI ; Ruiting BAI ; Chen MA ; Yuyu ZHANG ; Yiqun GUO ; Lin WU ; Shufan SONG ; Cuilian LI
Chinese Journal of Laboratory Medicine 2009;32(1):51-54
Objective To analyze the application value of the rapid testing for influenza during 2007-2008 flu season at fever clinic in Beijing Chaoyang hospital Methods 500 patients with diagnosis of influenza-like illness were prospectively enrolled. Pharyngeal swabs were collected for influenza viral culture and rapid testing for influenza. Demographic characteristics, age, symptoms, lab tests, symptom recovery time and medical expense were also collected. The sensitivity, specificity, positive predictive value and negative predictive value for rapid testing were analyzed. Results A total of 500 patients were enrolled between Dec 2007 and March 2008. Among them 498 cases were used for analysis. Influenza B was most common by virus culture methed(n=208,41.8%) ,followed by influenza A (n=51,10.2%). The average age was 35, and the ratio of male to female was 1.47:1. Compared with the group of positive culture, patients with influenza were more likely to get cough, sore throat, and nasal congestion (t=13.728, 4.014and 4.720,P<0.001 or 0.05, respectively). A total of 260 cases were subjected to rapid testing, Among them 18 cases were influenza A positive and 132 cases were influenza B positive. The rapid testing had a sensitivity of 77.1 % and a specificity of 70.1%. The positive predictive value was 78.6% and the negative predictive value was 68.2%. The rapid testing had enhanced the proportion of anti-viral treatment from 0 to 26% and reduced the proportion of antibiotic use from 63.4% to 20. 7%. Conclusions Influenza B is the most predominant pathogen during 2007-2008 flu season among patients with influenza-like illness in Beijing. The rapid testing with high sensitivity and specificity provides guidance on clinical practice.
3.Effects of Shizidaiping formula on MIN6 cell apoptosis and expressions of MEK1/2 and ERK1/2
Ruiting CHENG ; Yushan DONG ; Jian LI ; Xiaojin LA ; Chunyu TIAN ; Xiujuan GAO ; Xuemei ZHOU ; Qianru FU ; Bo WU ; Siqi DONG
Chinese Journal of Tissue Engineering Research 2017;21(4):603-608
BACKGROUND:Apoptosis of islet cel s is closely related to the long-term hyperglycemia-and hyperlipemia-induced injuries. OBJECTIVE:To observe the effect of Shizidaiping formula on the apoptosis and insulin secretion in MIN6 cel s under the high glucose and lipid environment, and to explore the protective effect of Shizidaiping formula and the related apoptosis mechanism. METHODS:MIN6 cel s were divided into normal, model, melbine, low-, medium-and high-dose Shizidaiping formula groups. The cel activity was examined by cel counting kit-8, the insulin secretion was measured by ELISA, the rate of apoptosis was measured by Annexin V-FITC&PI and the expression levels of MEK1/2, ERK1/2 and p-ERK1/2 were examined by western blot assay. RESULTS AND CONCLUSION:Shizidaiping formula significantly improved MIN6 cel activity under high glucose and lipid condition (P<0.05), decreased early cel apoptosis, increased the level of insulin stimulated by low glucose in cel supernatant (P<0.05), and improved the expression levels of MEK1/2, ERK1/2 and p-ERK1/2 (P<0.05). These results suggest that Shizidaiping formula can protect islet cel s from hyperglycemia and hyperlipemia damage by improving the activity of MIN6 cel s, reducing the insulin secretion and inhibiting the apoptosis of pancreaticβcel s in MIN6 cel s.
4.Mechanism of malignant behavior in cervical cancer cells based on NOD2-mediated AMPK/mTOR signaling pathway
Ruiting DU ; Dongyue WU ; Qingmin GUO ; Dongmei JIN
Acta Universitatis Medicinalis Anhui 2024;59(2):316-324
Objective To explore the mechanism of malignant behavior of cervical cancer(CC)cells based on AMP-activated protein kinase(AMPK)/rapamycin target protein(mTOR)signaling pathway mediated by nucleo-tide-binding oligomerization domain receptor 2(NOD2).Methods Bioinformatics analysis was performed to deter-mine the expression of NOD2 in CC tissue.Plasmids targeting NOD2(shNOD2)and shRNAs negative control(shNC),NOD2 overexpression(NOD2)and vectors(Vec)were transfected into CC cells.The effect of NOD2 on the growth of CC cells was determined by cell counting kit-8 assay,colony formation and Transwell cell invasion as-say.Transcriptome analysis was performed by high throughput RNA sequencing(RNA-Seq).Western blot was used to detect the expression of NOD2,AMPK/mTOR signaling pathway and autophagy protein in the cell line.24 female BALB/c nude mice were randomly divided into four groups,with 6 mice in each group:vector group(Vec group),NOD2 overexpression group(NOD2 group),shNC group and shNOD2 group.The distant metastasis mod-el was established in mice,and the fluorescence intensity of lung metastasis was monitored and the number of lung metastasis nodules was counted.Results On-line database analysis showed that the expression of NOD2 in CC tis-sues was significantly higher than that in normal tissues,and there were significant differences in the mRNA expres-sion of NOD2 in different stages of CC(P<0.05).In addition,the high expression of NOD2 was associated with poor overall survival and disease-free survival(P<0.05).NOD2 overexpression promoted the proliferation,colony formation,migration and invasion of CC cells,while NOD2 knock-down was the opposite.Consistent with the re-sults in vitro,the lung colonization and lung metastasis of CC cells in NOD2 group were significantly higher than those in Vec group(P<0.05),while those in shNOD2 group were significantly lower than those in shNC group(P<0.05).RNA-Seq results showed that the expression of NOD2 was significantly related to AMPK signal activa-tion,mTOR signal inhibition,autophagy regulation pathway activation and autophagy formation.Compared with shNC group,the expression of phosphorylated AMPK and LC3 protein decreased significantly in shNOD2 group(P<0.05),and the expression levels of phosphorylated mTOR and p62 protein increased significantly(P<0.05).Compared with Vec group,the expression levels of LC3 and AMPK protein in NOD2 group increased significantly(P<0.05),and the expression levels of phosphorylated mTOR and p62 protein decreased significantly(P<0.05).Compared with shNC group,the point accumulation of GFP-mRFP-LC3 in shNOD2 group decreased signifi-cantly(P<0.05).Compared with Vec group,the point accumulation of GFP-mRFP-LC3 increased significantly(P<0.05).Conclusion NOD2 may promote the proliferation,migration and invasion of CC through AMPK/mTOR signal,and its mechanism partly involves autophagy activation.
5.Construction of a hypoglycemia prediction model for older adults with type 2 diabetes based on random forest algorithm
Ruiting ZHANG ; Yu LIU ; Aiqing HAN ; Quanying WU ; Jing WANG ; Jingyi LIU ; Xiaoyan BAI
Chinese Journal of Practical Nursing 2023;39(23):1829-1835
Objective:To construct a hypoglycemia random forest prediction model for older adults with type 2 diabetes, and assess the model′s prognostication performance through internal and external verification.Methods:From August 2022 to January 2023, 300 older adults with type 2 diabetes in Beijing Hospital were selected. The demographic characteristics, medical history, laboratory tests, and other data of the patients were collected, and the data set was randomly divided into the training set and verification set in a ratio of 7∶3. The hypoglycemia prediction model for older adults with type 2 diabetes was constructed and optimized based on the random forest algorithm. The calibration curve was used to evaluate the model′s calibration, and the ROC was used to evaluate the model′s discrimination. The clinical applicability of the model was assessed by the decision curve analysis. The risk factors for hypoglycemia in the older adults were explored by prioritizing the contributions of variables in prediction. The Bootstrap method was used for internal validation, and the validation set was used for external validation.Results:Among the 300 older adults with type 2 diabetes, 128 cases (42.67%) experienced hypoglycemia within one week. The predictive contributions of risk factors in the model were ranked as follows: the number of episodes of hypoglycemia in one month, HDL-C, heart disease, diabetes knowledge and education, combination therapy, age, duration of diabetes, staple food restriction, glycosylated hemoglobin, and gender. The internal and external calibration curves of the hypoglycemia random forest model for the older adults with type 2 diabetes fluctuated around the diagonal, indicating that the calibration degree of the predictive model is good. The AUROC of internal verification was 0.823 (95% CI 0.752-0.894), the sensitivity and specificity were 0.867 and 0.698, respectively. The external verification was 0.859 (95% CI 0.817 - 0.902), and sensitivity and specificity were 0.789 and 0.804, respectively, showing that the overall discrimination of the prediction model was good. The DCA curves were far from the all-positive line and all-negative line, which indicated that the prediction model had good clinical applicability. Conclusions:The predictive effect of this model is good, and it is suitable for predicting the risk of hypoglycemia in older adults with type 2 diabetes, and it provides a reference for early hypoglycemia screening and predictive intervention for this kind of patients.
6.Inhibitory effect of Pingwei capsule on oxidative stress in GES-1 cells af-ter malignant transformation and its mechanism
Lijuan WANG ; Longde WANG ; Xiaoying NIU ; Xia WANG ; Ruiting ZHANG ; Yuqian WU ; Zekun FAN
Chinese Journal of Pathophysiology 2024;40(1):58-67
AIM:To observe the effect of Pingwei capsule on the precancerous lesions of gastric cancer(PLGC)cell model induced by N-methyl-N'-nitro-N-nitrosoguanidine(MNNG),and to preliminarily explore its mecha-nism.METHODS:Blank serum and Pingwei capsule-containing serum were prepared for later use.A PLGC cell model was established by MNNG-induced human gastric mucosal epithelial cell line GES-1.To evaluate the model,cell morpho-logical changes were observed under inverted microscope,and the expression of proliferating cell-related antigen Ki67 was detected by immunofluorescence staining.CCK-8 assay was used to screen the optimal intervention concentration and time of serum containing drugs.Reactive oxygen species(ROS)content in cells was detected using a fluorescent probe DCFH-DA.Malondialdehyde(MDA)content was detected by ELISA,and the activity of superoxide dismutase(SOD)and gluta-thione peroxidase(GSH-Px)was detected using biochemical reagents.A novel fluorescent probe JC-10 was used to detect mitochondrial membrane potential.The mRNA expression levels of Ki67 and melanoma differentiation-associated gene-7(MDA-7)were detected by real-time fluorescence quantitative PCR.The protein expression levels of Ki67,interleukin-6(IL-6)and MDA-7 were detected by Western blot.RESULTS:Compared with normal group,the ROS and MDA levels in model group and blank serum group were significantly increased(P<0.01),while the activity of SOD and GSH-Px was significantly decreased(P<0.01).The mitochondrial membrane potential was significantly decreased(P<0.01).The protein expression levels of Ki67 and IL-6 were significantly increased(P<0.01),while the protein expression level of MDA-7 was significantly decreased(P<0.01).There were no significant differences of the above indicators between model group and blank serum group(P>0.05).Compared with blank serum group,the Pingwei capsule-containing serum group showed significantly decreased ROS and MDA levels(P<0.01),significantly increased activity of SOD and GSH-Px(P<0.05),significantly increased mitochondrial membrane potential(P<0.01),significantly decreased protein expres-sion levels of Ki67 and IL-6(P<0.01),and significantly increased protein and mRNA expression levels of MDA-7(P<0.01).CONCLUSION:Pingwei capsule can significantly alleviate MNNG-induced oxidative damage and inflammatory response,and regulate the expression of oncogenes and tumor suppressor genes,thereby playing a role in prevention and treatment of PLGC.
7.Research progress in the role of M1/M2 polarization of macrophages in different liver diseases
Yuanyuan NIU ; Longde WANG ; Wenjuan XU ; Zhengju LI ; Ruiting ZHANG ; Yuqian WU
Journal of Shanghai Jiaotong University(Medical Science) 2024;44(4):509-517
Macrophages have strong plasticity and heterogeneity,and can undergo functional transformation in response to different signal stimuli,such as classical activation of M1 type(M1 type polarization)and selective activation of M2 type(M2 type polarization).The pathways of macrophage M1/M2 polarization are quite extensive,involving nuclear factor-κB(NF-κB)/mitogen-activated protein kinase(MAPK)signaling pathway,interleukin-4(IL-4)/signal transduction and activator of transcription 6(STAT6)signaling pathway,Notch signaling pathway,Wnt/β-catenin signaling pathway,etc.At the same time,M1/M2 polarization of macrophages is also regulated by exosomes,metabolites,non-coding RNA,electrical stimulation,probiotics,etc.,and its imbalance is closely related to the occurrence and development of different types of liver disease.In this paper,the mechanism of its polarization was reviewed,and it was found that M1 polarization of macrophages played a promoting role in the process of liver tissue injury,inflammation and fibrosis,while M2 polarization of macrophages played the opposite role.Among them,hepatocellular carcinoma,as the advanced stage of chronic liver disease,was characterized by increased M2 polarization and impaired M1 polarization of macrophages.Therefore,this paper pays attention to the role of M1/M2 polarization of macrophages in different types of liver diseases,in order to better establish the targeted therapy of macrophage subsets.
8.Research progress of theory selection tools in the field of implementation science
Ruiting ZHANG ; Yu LIU ; Yufang HAO ; Shishi WU ; Xue QIAO
Chinese Journal of Modern Nursing 2022;28(13):1681-1686
In the field of implementation science, the use of appropriate theories is an important basis for promoting the promotion and application of evidence-based practice. However, the specific methods of selecting theories in the research design are rarely mentioned. This paper reviews the main contents, usage methods, characteristics and applications of "Theory Comparison and Selection Tool" (T-CaST) and A Pragmatic Guide for Selecting Theoretical Approaches in the field of implementation science, aiming at providing a reference for researchers to select appropriate theories.
9.Effect of lentivirus vector-mediated RNA interference dbpA gene silencing on the biological behavior of colorectal cancer cells
Ruiting LIU ; Yali HOU ; Xiangtian WU ; Guorong WANG ; Chang LIU ; Jirong BAI ; Jian QIU ; Likun YAN ; Xiaojun LI ; Xiaoqiang WANG
Chinese Journal of General Surgery 2019;34(7):613-617
Objective To investigate the effects of lentivirus-mediated RNA interference (RNAi) targeting DNA binding protein A (dbpA) on the proliferation and the biological behavior of colorectal cancer cell line SW620.Methods The experiment was divided into 3 groups:KD group (siRNA-dbpA,lentivirus interference group),CON group (non-specific sequence group) and NC group (blank control group).The lentiviral vector siRNA-dbpA was constructed and verified by PCR and DNA sequencing.SW620 cells were transfected with siRNA-dbpA plasmid,nontargeting siRNA plasmid,or empty plasmid.After 48 h the transfection,the cells were examined for dbpA expression using Western blot.After 72 hrs transfection,flow cytometry was used to detect the cell apoptosis and cell cycle changes.The cell growth inhibition rate was detected by MTT (4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide) assay,and then clone formation was detected,and the ability of SW620 cells to form tumors in vivo after dbpA was silenced was studied in nude mice.Results PCR analysis and DNA sequencing demonstrated that the RNAi sequence targeting dbpA gene was successfully inserted into the lentiviral vector.siRNA-dbpA transfection resulted in reduced expression of dbpA in SW620 cells.After transfection,the apoptosis rate of siRNA-dbpA-transfected cells increased to 26.60% ± 0.38%,significantly higher than that in cells transfected with the nontargeting plasmid or the empty plasmid 12.54% ± 0.25% and 4.46% ± 0.19%,respectively (F =28.159,P <0.01).The growth inhibition test indicate that the OD value of the fifth day in siRNA-dbpA group was 0.194 ±0.037,significantly lower than that in the other two groups 0.814 ±0.043 and 1.625 ±0.061,respectively(F =23.214,P < 0.01).The colony formation number is 37 ± 3,64 ± 5and 175 ± 10 respectively,siRNA-dbpA is significantly higher than that in the other two groups(F =40.254,P < 0.01).After the completion of nude mouse transplantation tumor model,through the detection of tumor volume,KD group (group siRNA-dbpA) tumor volume after 14 d and CON and NC group had obvious difference (F =38.256,P < 0.05),and after 21d is more significant difference in tumor size (F =40.241,P < 0.01),can be clearly observed after 35 d KD group (group siRNA-dbpA) growing tumors had differences with the control group (F =30.257,P < 0.05).Conclusion Lentivirus-mediated RNAi targeting dbpA can effectively suppress the expression of dbpA in colorectal tumor in nude mice,it is proved that dbpA silencing has a significant inhibitory effect on the growth of living tumor cells and decrease the proliferation of the colorectal cells.
10.Analysis of pathogenic gene variant in two children with Treacher-Collins syndrome
Jie WANG ; Xiaoping JI ; Lichun ZHANG ; Ruiting XU ; Yan HUANG ; Yaxian LIU ; Liqiong WU ; Jin AN ; Zhiyuan GUO ; Xiaohua WANG
Chinese Journal of Medical Genetics 2022;39(6):625-629
Objective:To explore the clinical and genetic characteristics of two children with a clinical diagnosis of Treacher Collins syndrome (TCS).Methods:Whole-exome sequencing was used to screen potential variants in the two children. Confirmation of suspected variants was performed through Sanger sequencing , multiplex ligation dependent probe amplification and real-time PCR in probands and their parents.Results:A heterozygous deletion variant, c. 4357_4360delGAAA, was detected in case one, while was de novo and verified by Sanger sequencing. Thevariant was classified as pathogenic(PVS1 + PM2+ PM6)according to ACMG guideline. The heterozygous deletion of exon 1-7 was seen in the same gene in case 2, which MLPA verified as heterozygous deletion of exon 1-6. This deletion was inherited from the father with a normal phenotype, and the father’s TCOF1 gene was suspected to be chimeric heterozygous deletion of exon 1-6 verified by MLPA. Conclusion:The identified variants in the TCOF1 gene probably underlie the two cases of TCS. There was no apparent correlation between genotype and phenotype. In addition, it shows a high interfamilial variability ranging from normal to full presentation of TCS. Genetic detection provided clinical diagnosis and genetic counselling for TCS patients .