2.Expression of CCRK in small cell lung cancer and its clinical significance
Zewei SHAO ; Fenglian SHAN ; Weiwei SUN ; Leisheng LI ; Shangdan NIE ; Xiaoxia ZHANG ; Hong ZHENG ; Qisen GUO
Journal of Chinese Physician 2015;(3):343-346
Objective To investigate the pathogenesis of small cell lung cancer ( SCLC) and to ex-plore the expression of cell cycle related kinase ( CCRK) in SCLC and its clinical significance.Methods Reverse transcription polymerase chain reaction ( RT-PCR) and Western blot were performed to examine ex-pression of CCRK in SCLC and normal tissues.Results The expressions of gene [(0.51 ±0.11)IU/L] and protein [(0.61 ±0.13)IU/L] of CCRK in SCLC tissues were significantly higher than normal tissues [(0.30 ±0.08)IU/L, (0.34 ±0.09)IU/L] ( P <0.05).The expression of CCRK was closely correlated with the clinical curative effect ( P <0.05 ) rather than the clinical stages ( P >0.05 ) .Conclusions The expressions of gene and protein of CCRK in SCLC tissues were significantly higher than normal tissues. CCRK promoted the occurrence and progress of SCLC.Chem can restrain effectually the excessive expres-sion of CCRK.The expressions of gene and protein of CCRK in the different clinical curative effect group had significant difference.
3.Reformation and exploration on laboratory teaching in bacteriological analysis
Haixia YANG ; Ting CHEN ; Leisheng LI ; Zewei SHAO ; Xiaoxia ZHANG ; Shangdan NIE
Chinese Journal of Medical Education Research 2005;0(05):-
The laboratory teaching of bacteriological analysis is very important for training practical analysis professionals.For better quality of laboratory teaching,in this paper,we practice a series of reform facilitating the lab teaching of bacteriological analysis,which is involved in the exploration and study of new curriculum system and teaching evaluation.The educational practices create effective teaching and learning environment and better performance.
4.Effect and mechanism of gene therapy of lentivirus mediated RhoA shRNA on ovarian cancer xenograft in vivo
Wenyan JIANG ; Jiali KANG ; Xiaoxia WANG ; Wenjuan YANG ; Miaoling NIE ; Cong ZHOU
Chinese Journal of Obstetrics and Gynecology 2013;48(10):778-783
Objective To investigate treatment effects of lentivirus mediated RhoA short hairpin RNA(shRNA) on xenograft tumor of ovarian cancer in nude mice in vivo and the underlying mechanism.Methods Human ovarian cancer cell line HO8910 were inoculated to establish subcutaneous xenograft model of human ovarian cancer.Tumor-bearing nude mice were assigned randomizely to three groups:LentiRhoA-sh group,Lenti-negative control (NC) group and phosphate buffered saline (PBS) group.lentivirus mediated RhoA shRNA,negative control lentivirus and PBS were respectively injected in the three groups.Effects of treatment were observed by tumor growth curve,tumor volume,tumor weight,and tumor inhibition rate.Xenograft tissues and liver,spleen,lung,and renal tissues were examined by hematoxylin and eosin (HE) staining or were detected by streptavidin-perosidase (SP)immunochemical method.The changes of RhoA gene expression in xenograft tissues after lentivirus mediated RhoA shRNA treated were also detected by real-time qPCR,immunochemistry and Western blot assay.Cell apoptosis in xenograft tissues were examined by terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling(TUNEL) method and apoptotic index (AI) were counted.Results Compared with Lenti-NC group and PBS group,the growth speed of xenograft in Lenti-RhoA-sh group delayed significantly after injection 9 days(P < 0.01).Tumor volume (338 ± 114) mm3 decreased significantly in the Lenti-RhoA-sh group when compared with those in Lenti-NC group(1190 ± 332)mm3 and PBS group (1101 ± 396) mm3 (P < 0.01).Tumor weight (0.23±0.11)g decreased significantly in the Lenti-RhoA-sh group when compared with Lenti-NC group (0.79 ± 0.19)g and PBS group (0.74 ± 0.17)g (P < 0.01).Real-time qPCR result shown that the expression of RhoA mRNA (0.30 ± 0.05) decreased significantly in the Lenti-RhoA-sh group compared with Lenti-NC group (0.95 ±0.06) and PBS group(1.00 ±0.11 ; P <0.01).Western blot result showed that the expression level of RhoA protein decreased significantly in the Lenti-RhoA-sh group (0.14 ± 0.06) compared with those in Lenti-NC group(0.78 ± 0.14) and PBS group (0.75 ± 0.13;P < 0.01).TUNEL staining displayed that AI significantly increased in the Lenti-RhoA-sh group (20.9 ± 3.4) % compared with those in Lenti-NC group(5.2 ±±2.0)% and PBS group(6.0 ±2.1)% (P <0.01).Conclusion Lentivirus mediated RhoA shRNA may be effectively down-regulate of the expression of RhoA,inhibit the growth of subcutaneous xenograft tumor of ovarian cancer in nude mice by increasing the cell apoptosis.
5.Establishment of TaqMan RT-PCR assay for Flanders virus
Hao LI ; Xiaoxia HE ; Yuxi CAO ; Kai NIE ; Yan LIU ; Ying HE ; Xiaoyan GAO ; Shihong FU ; Huanyu WANG
Chinese Journal of Zoonoses 2015;(3):212-215
The Flanders virus (FLAV) is a number of family Rhabdoviridae ,contains a single‐stranded ,negative‐sense vi‐ral RNA .Here we describe a molecular detection method developed for fast measurement of FLAV based on Taqman RT‐PCR method .In this study ,FLAV specific primers and probe were designed based on the FLAV L gene sequences published in GeneBank .Quantitative standard curve of FLAV TaqMan PCR was also successfully established .The specificity and stability test showed that the system is specific and the coefficient variables were all less than 1 .7% .Quantitative standard curve based on the genomic copy was drawn ,and the lowest detectable limit (LOD) of system was 100 copies/PCR ,with higher sensitivity and stability than that of the conventional RT‐PCR assay targeting the same gene .
6.The detection of relationship between cellular immune function and CD4 + T lymphocytes apoptosis in patients with pulmonary tuberculosis
Ting CHEN ; Shangdan NIE ; Yuexia ZHAO ; Xiaoxia ZHANG ; Haixin DONG ; Yuanming ZHAO ; Zhuoxiang SUN ; Xinke CHEN ; Leisheng. LI
Journal of Chinese Physician 2011;13(12):1595-1598
ObjectiveTo study the apoptosis of CD4 + T lymphocytes and the detection of immune function in patients with pulmonary tuberculosis and explore the clinical significance.MethodsThe mononuclear cells were separated from the blood of the tuberculosis patients or the healthy.The flow cytometry was used to measure the percentage of apoptotic CD4 + T lymphocytes,and the standard of T-lymphocyte subsets were detected by using SAP technology.The red cell immune function were determined by using yeast wreath way.Results The apoptosis rate of CD4 + T lymphocytes and CD8 + T lymphocyte was ( 15.882 ± 4.65 ) %,and (27.69 ± 0.74) %.The Immune complex positive rate ( 19.40 ± 0.58) % in patients with tuberculosis was significantly higher than those in controls ( P < 0.01 ).C3b receptor positive rate in red blood cells was ( 17.73 ± 0.63 ) %,( 46.48 ± 1.34 ) % in CD3 + T lymphocyte,( 28.12 ±0.69 ) % in CD4 + T lymphocyte,and the ratio of CD4/CD8 ( 1.0223 ± 0.09362) in the patients with tuberculosis was lower than the control group( P < 0.01 ).There were certain relationships between the apoptosis rate of CD4 + T lymphocytes and the percentages of CD4 + T lymphocyte,the standard of T lymphocyte subsets and the red cell immune function.ConclusionsThe apoptosis rate of CD4 + T lympho,cytes in patients with tuberculosis were significantly higher than the healthy,which led to reducing the number of CD4 + T lymphocytes.There was positive correlation between red cell immunity and T-lymphocyte immunity,and the immunity in red cell and T- lymphocyte was lower than normal controls,which may be related to the immune pathogenesis of pulmonary tuberculosis.
7.Prokaryotic expression, purification, and identification of recombinant human IL-11 protein.
Jia TANG ; Xiaoling XU ; Xiaoxia NIE ; Qifeng MAO ; Jimin GAO
Journal of Biomedical Engineering 2012;29(3):530-533
A DNA fragment encoding recombinant human interleukin 11 (hrIL-11) was obtained by PCR from previously constructed pET24a-hrIL-11 plasmid. Then pET21a-hrIL-11 expression vector was constructed routinely and transformed into BL-21(DE3). By the induction of Isopropyl-1-thio-beta-D-galactoside (IPTG), hrIL-11 protein was highly expressed at about 20% of the total bacterial proteins and was identified by Western blot. After purification with Ni-NTA affinity chromatography and refolding with renaturation buffer, the purity of the target hrIL-11 protein reached 95% and its biology activity was 1 x 10(6) IU/mg, determined by stimulating the proliferation of T1165, which facilitates further researches into effects of IL-11 on platelet proliferation and other function.
Escherichia coli
;
metabolism
;
Genetic Vectors
;
genetics
;
Humans
;
Interleukin-11
;
genetics
;
metabolism
;
Recombinant Proteins
;
genetics
;
isolation & purification
;
metabolism
8.Effect of dexmedetomidine mixed with dexamethasone on efficacy of ropivacaine for popliteal sciatic nerve block in patients undergoing ankle surgery
Xiaona WANG ; Zhixue WANG ; Chong LIU ; Long DONG ; Hanxiao NIE ; Deli ZHANG ; Xiaoxia ZHANG
Chinese Journal of Anesthesiology 2020;40(5):600-602
Objective:To evaluate the effect of dexmedetomidine mixed with dexamethasone on efficacy of ropivacaine for popliteal sciatic nerve block in the patients undergoing ankle surgery.Methods:A total of 120 patients of either sex, aged 30-64 yr, with body mass index of 19.6-29.7 kg/m 2, of American Society of Anesthesiologists physical status Ⅰ or Ⅱ, undergoing elective ankle surgery, were divided into 4 groups ( n=30 each) by a random number table method: control group (group C), dexmedetomidine group (group DD), dexamethasone group (group DM), and dexmedetomidine plus dexamethasone group (group DD+ DM). In group C, 0.5% ropivacaine 30 ml was injected around the popliteal sciatic nerve guided by ultrasound combined with a nerve stimulator.Dexmedetomidine 1 μg/kg, dexamethasone 10 mg and dexmedetomidine 1 μg/kg plus dexamethasone 10 mg were added to 0.5% ropivacaine in group DD, group DM and group DD+ DM, respectively.The analgesic time, consumption of sufentanil and adverse reactions were recorded after popliteal sciatic nerve block. Results:Compared with group C, the analgesic time was significantly prolonged, the consumption of sufentanil was reduced, and the incidence of nausea and vomiting was decreased in group DD, group DM and group DD+ DM ( P<0.05). Compared with group DD and group DM, the analgesic time was significantly prolonged, and the consumption of sufentanil was reduced in group DD+ DM ( P<0.05). No itching, drowsiness, hypotension, bradycardia or respiratory depression occurred in each group. Conclusion:Dexmedetomidine mixed with dexamethasone can effectively enhance the efficacy of ropivacaine for popliteal sciatic nerve block in the patients undergoing ankle surgery.
9.Analysis of public demand for information related to congenital birth defects in “Baidu know” based on word frequency of internet retrieval
Zehao WU ; Zimo SUN ; Yaguang PENG ; Xiaolu NIE ; Siyu CAI ; Xiaoxia PENG
Chinese Journal of Health Management 2021;15(3):237-242
Objective:To analyze the public demands for information about congenital birth defects in “Baidu zhidao” based on word frequency retrieval.Methods:Based on discussion between obstetrics and gynecology experts and epidemiological experts, the key words related to congenital birth defects were determined and the search strategy was formulated. Python 2.7 was used for web crawler search. Questions related to congenital birth defects were obtained on the “Baidu zhidao” platform, and then the R 4.0.2 software was used to process the data, complete the semantic analysis of keywords and statistical analysis of word frequency, and draw word cloud graph and polar chart to describe the key results.Results:A total of 16668 non-repetitive questions were retrieved from “Baidu zhidao” platform, and the frequency of semantic words was 15 371. Among them, 35.02% were the names and symptoms of congenital birth defects. In addition, the frequency of congenital heart disease was the highest (26.09%). The results of subject analysis of key words of birth defects showed that the average word frequency of diagnosis and treatment semantic words (49.55) was significantly higher than that of etiology and prevention semantic words (12.47). In addition, the key words of examination, cause, treatment, development and heredity were more frequently used in the semantic words related to the seven types of systemic malformations.Conclusion:The public in China has a high demand for information on congenital birth defect related diseases, and their causes, prevention and treatment, especially congenital heart disease.
10.Development and validation of a diagnostic model based on machine learning algorithms for the development of interstitial lung diseases in patients with rheumatoid arthritis
Yancong NIE ; Yanqing JIN ; Meilin YIN ; Xiaoxia WANG ; Lixia QIU
Chinese Journal of Rheumatology 2024;28(3):167-175
Objective:Screening factors that might influence rheumatoid arthritis (RA) complicating interstitial lung diseases (ILD) by constructing and validating a model for early diagnostic.Methods:The study subjects were composed of 712 RA patients in the Department of Rheumatology and Immunology of the Second Hospital of Shanxi Medical University during December 2019 to October 2022. Fifty-two variables such as their demographic data, clinical symptoms, and laboratory indexes were collected. Patients were categorized into RA-only group and RA-ILD group with or without the occurrence of ILD disease. After data preprocessing, subjects were randomly assigned to the modeling and validation groups in a 7:3 ratio.Univariate analysis comparing baseline characteristics of the two groups of patients. Feature selection was performed using LASSO and SVM-RFE regression algorithms.Screening indicators were analyzed by logistic regression and the results were used to develop a nomograms model for the early diagnosis of RA complicating interstitial lung disease; and the modeling group was evaluated for its performance for internal assessment of the model and internal validation using data from the validation group.Results:A total of 712 subjects participated in the study, of which 498 in the modeling group and 214 in the validation group. Univariate analysis showed that the differences between the two groups were statistically significant ( P<0.05) in 18 characteristic indexes, including male, gender, age, smoking history, drinking history, number of swollen joints, number of painful joints, use of prednisone, WBC, ESR, CRP, IL-2, IL-10, IL-17, TNF-α, INF-γ, AFA family, APF, and serum albumin. The LASSO algorithm identified 13 risk variables for RA-ILD, the SVM-RFE algorithm identified 12 variables for RA-ILD, and the intersecting risk variables were male, age, history of alcohol consumption, number of painful joints, prednisone acetate, IL-2, AFA family, TNF-α, serum albumin, and IL-10. The results of multifactorial logistic regression analysis confirmed that the differences between males [ OR(95% CI)=3.61(2.11, 6.18)], gender, age [ OR(95% CI)=1.05(1.03, 1.08)], number of painful joints [ OR(95% CI)=1.03(1.01, 1.06)], IL-2 [ OR(95% CI)=0.91 (0.84, 0.99)], and TNF-α[ OR (95% CI)=1.06 (1.02, 1.10)] were statistically significant ( P<0.05) and were independently influences on ILD complicated by RA. The modeling and validation groups that were used to construct early diagnostic Nomograms had high calibration curve accuracies, and the model had a high diagnostic power, which was mainly demonstrated by the receiver operating characteristic (ROC) area under the curve (AUC) and decision curve analysis(DCA), the model modeling group had an AUC of 0.76 (95% CI=0.71, 0.81), with net benefit rates of 3%~82% and 93%~99%, whereas the model validation group had an AUC of 0.71 (95% CI=0.64, 0.79), with net benefit rates of 5%~11%, 14%~60% and 85%~89%. Conclusion:Male, gender, age, number of painful joints, IL-2, and TNF-α are independent factors for RA complicated with ILD, and the Nomogram model constructed has good performance in early diagnosis of the disease.