Objective To construct and evaluate the real-time fluorescence quantitative polymerase chain reaction for detecting IL-2R? mRNA in ankylosing spondylitis(AS) based on TaqMan technique.Methods A pair of primers and a TaqMan probe were designed by sequence in GenBank.Total RNA isolated from the fresh peripheral blood monocytes (PBMC) of homo sapiens was amplified by the real-time FQ-RT-PCR.The product was collected by agarose gel electrophoresis and sequencing analysis then ligated with pUCm-T.The recombined plasmid was transcribed to cRNA by T7 RNA polymerase in order to prepare serial standard materials.A new method was created to quantify IL-2R? mRNA in ideal condition.Sensitivity, reproducibility and efficiency were evaluated and used, combined with sIL-2R,for clinical application of AS.Results The linear range was (7~107) cells/ml.The intra-and-inter-assay coefficient variation was 8.4% and 9.6% respectively. Recombined plasmid contained the target fragment was specific and accurate by BLAST.Standard reference was constructed successfully.The RT-PCR product in AS with HLA-B27 positive groups was higher than that with HLA-B27 negative groups and health controls (P0.05).The sensitivity of IL-2R? mRNA was 96.7%.Conclusions Real Time FQ-RT-PCR of IL-2R? mRNA is constructed successfully.This is an easy,rapid,sensitive, accurate and reliable method for quantifing IL-2R? mRNA. There is highly statistical significance, compared with sIL-2R, on the expression of IL-2R? mRNA and inflammatory states between AS and control group and effective information for administration of patients.

[Objective]Observe the clinical effect of leverage replacement manipulation on lumbar intervertebral disc herniation.[Method]299 hospitalized patients were divided into two groups.153 cases were in trial group and 146 patients were in controlled group.Trial group accepted constant traction and leverage replacement manipulation.While the controlled group accepted the same traction and muscular massage.[Result]In trial group,90 patients were cured,57 improved and 6 no improvement.The total effective rate was 96.08%;in controlled group,21 were cured,75 cases improved,and 50 no improvement.The total effective rate was 65.75%.The effect of trial group was better than the controlled group(P

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.

AIM:To develop and validate a predic-tive model for the risk of high plasma concentra-tion of voriconazole,and to guide clinical individual-ized medication of voriconazole.METHODS:Based on the real-world data from the hospital Informa-tion system(HIS),the clinical data of hospitalized patients who received voriconazole treatment and underwent voriconazole plasma concentration monitoring in our hospital from August 2017 to Au-gust 2021 were collected.Univariate and multivari-ate logistic regression analysis were performed on the included influencing factors.At the same time,in order to minimize the potential collinearity and overfitting between variables,the least absolute shrinkage and selection operator regression were used to screen the potential predictors.Logistic re-gression analysis was used to construct a predic-tion model for the risk of high plasma concentra-tion of voriconazole.C-index,calibration chart and clinical decision curve analysis were used to evalu-ate the discrimination,consistency and clinical ap-plicability of the model,and a nomogram was drawn.RESULTS:A total of 147 patients were en-rolled in this study.Plasma albumin and procalcito-nin were selected as predictive variables for Logis-tic regression analysis,and the prediction model was established.Draw predict voriconazole nomo-gram risk blood drug concentration on the high side.The receiver operating characteristic curve showed that the AUC of the prediction model for predicting the risk of high plasma concentration of voriconazole was 0.787(95%CI 0.663-0.911).Vori-conazole blood drug concentration was high inci-dence of cut-off value was 33.06%,sensitivity was 63.64%,87.65%and 58.33%positive predictive val-ue,negative predictive value of 89.87%.The cali-bration curve showed good consistency,and the clinical decision curve showed that the model had a positive net benefit when the threshold probabili-ty was between 6.67%and 99.99%.CONCLUSION:The predictive model for the risk of high plasma concentration of voriconazole has good predictive efficacy,which can provide guidance for clinical in-dividualized medication of voriconazole.