Objective: To prepare and purify siRNA targeting a proliferation-inducing ligand targeted (APRIL-siRNA), so as to provide a basis for studying the role of APRIL in human pancreatic cancer. Methods: pET-22b-APRIL was constructed to express APRIL dsRNA of human pancreatic cancer cell line CFPAC-1 in E. coli and the product was purified by chromatography using CF-11 column. APRIL dsRNA was digested by RNase III to prepare APRIL siRNA, then the reaction mixture was loaded onto a DEAE ion exchange chromatography to remove RNase III from oligonucleotides, and size exclusion chromatography was used to purify 21 bp siRNA. The purified APRIL siRNA was used to transfect Chinese hamster ovary (CHO) cells and the expression of APRIL in CHO cells was observed under fluorescence microscope. Results: APRIL dsRNA was successfully expressed in E. coli after IPTG induction and was purified by CF-11 column. dsRNA was hydrolyzed with RNase III and was purified by DEAE ion exchange chromatography and size exclusion chromatography. 15% nondenaturing PAGE and 12% SDS-PAGE confirmed that RNase III was removed from oligonucleotides and 21 bp siRNA was purified with size exclusion chromatography. It was also found that APRIL siRNA obviously depressed APRIL expression in CHO cells. Conclusion: We have successfully constructed APRIL siRNA targeting APRIL gene of CFPAC-1 cells with in vitro transcription, which provides a basis for knock-down of APRIL gene in CFPAC-1 cells.

Objective:To prepare and purify siRNA targeting a proliferation-inducing ligand targeted(APRIL-siRNA),so as to provxde a basis for studying the role of APRIL in human pancreatic cancer.Methods:pET-22b-APRIL was constructed to express APRIL dsRNA of human pancreatic cancer cell line CFPAC-1 in E.coli and the product was purified by chromatography using CF-11 column.APRIL dsRNA was digested by RNaseⅢto prepare APRIL siRNA,then the reaction mixture was loaded onto a DEAE ion exchange chromatography to remove RNaseⅢfrom oligonucleotides,and size exclusion chromatography was used to purify 21 bp siRNA.The purified APRIL siRNA was used to transfect Chinese hamster ovary(CHO)cells and the expression of APRIL in CHO cells was observed under fluorescence microscope Results:APRIL dsRNA was successfully expressed in E.coli after IPTG induction and was purified by CF-11 column.dsRNA was hydrolyzed with RNaseⅢand was purified by DEAE ion exchange chromatography and size exclusion chromatography.15% nondenaturing PAGE and 12% SDS- PAGE confirmed that RNaseⅢwas removed from oligonucleotides and 21 bp siRNA was purified with size exclusion chromatography.It was also found that APRIL siRNA obviously depressed APRIL expression in CHO cells.Conclusion:We have successfully constructed APRIL siRNA targeting APRIL gene of CFPAC-1 cells with in vitro transcription,which provides a basis for knock-down of APRIL gene in CFPAC-1 cells.

Objective To explore the feasibility of evaluating the lung function by MSCT in emphysema.Methods The MSCT scan and pulmonary function tests(PFF)were respectively performed in 147 receptors within one week.They were randomly divided into 2 groups:group A(120 receptors), including normal,mild,moderate and severe abnormal pulmonary function based on the PFT,for comparing the correlation between pulmonary quantitative indexes of MSCT pulmonary function and PFT and settingup the primary grade criteria of abnormal pulmonary function in emphysema,group B(27 receptors)for evaluating the diagnostic accuracy in group A.The total lung was respectively scanned at the full inspiration and full expiration with MSCT.The pulmonary quantitative indexes of MSCT were measured with Siemens Pulmo pulmonary quantitative software.Results There was correlation between pulmonary quantitative indexes of MSCT and PFF.The Piex/in_(-910)showed best correlation with FEV_1%(r=-0.905,P

OBJECTIVETo study the correlation between the lung pixel indexes of multi-slice spiral CT (MSCT) and pulmonary function test (PFT), and to explore the CT features of histograms of lung attenuation.

METHODSMSCT scan and pulmonary function test (PFT) were separately performed in 127 subjects within one week. The total lung was scanned with MSCT at full inspiration, full expiration, and normal respiration. The lung pixel indexes were measured by Pulmo pulmonary quantitative software. These indexes include PI(-910Huin), PI(-960HUEX), PI(-910Huin), PI(-960Huex), PI(-910(Hun), PI(-960Hun), PI(910Huex/in), and the histograms of lung attenuation.

RESULTSCorrelation existed between all the lung pixel indexes of MSCT and PFT. PI(-910Huex/in showed the best correlation with FEV1/FVC (r = 0.901, P < 0.01). Three distribution patterns were found in the histograms of lung attenuation of total lung, including double-peak distribution (n = 28), similar normal distribution (n = 81), and partial distribution (n = 18).

CONCLUSIONThe lung pixel indexes of MSCT have significant correlation with PFT and can be used to assess the pulmonary function.

Adult ; Aged ; Aged, 80 and over ; Female ; Humans ; Lung ; diagnostic imaging ; Male ; Middle Aged ; Respiratory Function Tests ; Tomography, Spiral Computed

Adult ; Aged ; Aged, 80 and over ; Diagnosis, Differential ; Humans ; Middle Aged ; Positron-Emission Tomography ; Retrospective Studies ; Solitary Pulmonary Nodule ; diagnosis ; Tomography, X-Ray Computed

OBJECTIVETo investigate the cut-off value in screening of thalassemia in pregnant women from Shenzhen region by capillary hemoglobin electrophoresis.

METHODSThe data of capillary hemoglobin electrophoresis and genetic diagnosis of thalassemia from 2122 examined prenatal women were retrospectively analyzed. Capillary hemoglobin electrophoresis and α-, β- genetic diagnosis of thalassemia were carried out for every woman. Hemoglobin electrophoresis was performed using Capillarys 2 full-automated electrophoresis instrument. Gap polymerase chain reaction and reverse dot blot were used for genetic diagnosis of thalassemia genotyping test. The cut-off value in screening of thalassemia was determined by receiver operating characteristic curve and next to analyze the value of HbA2 and HbF in screening of thalassemia using the decided cut-off value.

RESULTSThe areas under the curve (AUC(Roc)) of HbA2 for diagnosis of α-, β- thalassemia were 0.75 and 0.981 respectively, and the AUC(Roc) of HbF for diagnosis of β-thalassemia was 0.787. When HbA2 ≤ 2.55 was taken as the cut-off value of HbA2 for diagnosis of α-thalassemia, the sensitivity, specificity, positive likelihood ratio (LR(+)) and negative likelihood ratio (LR(-)) were 89.5%, 54.8%, 1.98, 0.19 respectively. When HbA2 ≥3.9 was taken as the cut off value of HbA2 for diagnosis of β-thalassemia, the sensitivity, specificity, LR(+) and LR(-) were 96.1%, 99.8% 480.5, 0.04 respectively. When HbF ≥0.75 was taken as the cut off value of HbF for diagnosis of β-thalassemia, the sensitivity, specificity, LR(+) and LR(-) were 83.6%, 61.8% respectively.

CONCLUSIONThe cut-off value in screening of thalassemia by capillarys 2 full automated electrophoresis instrument is different from that of the traditional method of hemoglobin electrophoresis, such as cellulose acetate membrane electrophoresis and agarose gel electrophoresis. Each laboratory should establish their own respective cut off value.

Area Under Curve ; China ; Electrophoresis, Capillary ; Female ; Fetal Hemoglobin ; analysis ; Genotyping Techniques ; Hematologic Tests ; Hemoglobin A2 ; analysis ; Humans ; Mass Screening ; Pregnancy ; Reference Values ; Retrospective Studies ; Sensitivity and Specificity ; alpha-Thalassemia ; diagnosis ; beta-Thalassemia ; diagnosis

During coronavirus disease 2019 epidemic, the treatment of severe trauma has been impacted. The Consensus on emergency surgery and infection prevention and control for severe trauma patients with 2019 novel corona virus pneumonia was published online on February 12, 2020, providing a strong guidance for the emergency treatment of severe trauma and the self-protection of medical staffs in the early stage of the epidemic. With the Joint Prevention and Control Mechanism of the State Council renaming "novel coronavirus pneumonia" to "novel coronavirus infection" and the infection being managed with measures against class B infectious diseases since January 8, 2023, the consensus published in 2020 is no longer applicable to the emergency treatment of severe trauma in the new stage of epidemic prevention and control. In this context, led by the Chinese Traumatology Association, Chinese Trauma Surgeon Association, Trauma Medicine Branch of Chinese International Exchange and Promotive Association for Medical and Health Care, and Editorial Board of Chinese Journal of Traumatology, the Chinese expert consensus on emergency surgery for severe trauma and infection prevention during coronavirus disease 2019 epidemic ( version 2023) is formulated to ensure the effectiveness and safety in the treatment of severe trauma in the new stage. Based on the policy of the Joint Prevention and Control Mechanism of the State Council and by using evidence-based medical evidence as well as Delphi expert consultation and voting, 16 recommendations are put forward from the four aspects of the related definitions, infection prevention, preoperative assessment and preparation, emergency operation and postoperative management, hoping to provide a reference for severe trauma care in the new stage of the epidemic prevention and control.