1.Feasibility study of a domestic fully automated NAT system for blood screening in blood donors
Fenglan YAO ; Rui WANG ; Jinghui HU ; Hongwei GE ; Chan LENG ; Yi ZHA ; Zifu ZHAO ; Zhengmin LIU
Chinese Journal of Blood Transfusion 2025;38(7):941-949
Objective: To validate the analytical performance, operational performance, and process control measures of a domestic fully automatic nucleic acid testing (NAT) system, thereby ensuring an efficient and orderly blood screening workflow. Methods: The concordance rate and sensitivity of WanTag-Vortex Plus system were verified using WHO standard reference panels of HIV-1, HCV and HBV, while precision was assessed using weak positive samples of HIV-1, HCV and HBV. As for its operational performance evaluation, cross-contamination resistance was assessed using strong positive samples, and throughput and stress testing were conducted using negative samples. Reagent stability was verified using weak positive samples, and inter-system performance consistency was assessed using verification panels. In addition, the process control measures were verified using the laboratory quality control demand scale. Results: 1) Verification of concordance rate: The detection results of negative and positive samples of HIV-1, HCV and HBV by WanTag-Vortex Plus system were all consistent with expectations, and the concordance rate was 100%. 2) Precision verification: the repeatability and intermediate precision were extremely high, and the coefficient of variation was less than 5%. 3) Verification of analytical sensitivity: The detection limit of 95% for standard strains of HIV-1, HCV and HBV by WanTag-Vortex Plus system in our laboratory was consistent with the analytical sensitivity provided by reagent manufacturers. 4) Verification of cross-contamination resistance: Five strong positive samples and 87 negative samples were placed according to the actual working conditions and equipment operation design, and the test results were consistent with expectations, with no cross-contamination in the testing system. 5) Throughput and stress testing: Each system completed the individual donor-nucleic acid amplification testing (ID-NAT) of 276 samples in three batches within 12 hours, and successfully completed the ID-NAT test of 828 samples in three consecutive days. 6) Verification of reagent stability: After extreme storage (unsealed storage for 1 week with 4 freeze-thaw cycles), the reagents maintained 100% detection rate in the weak positive samples of HIV-1, HCV, and HBV, showing no significant differences from the control group (Kappa=1). 7) Verification of inter-system performance consistency: The system has stable operation performance, and the performance comparison results across the four devices were consistent (Kappa=1). 8) Process control measures: WanTag-Vortex Plus system software accurately controlled the equipment operation process with strict quality control measures, and correctly interpreted and safely reported the test results. Conclusion: The analytical and operational performance of the WanTag-Vortex Plus system complies with manufacturer design standards and essential laboratory workflow requirements. Integrated with laboratory information system (LIS), the system's control software meets standard process control requirements, yet requires further improvement.
2.Feasibility study of a domestic fully automated NAT system for blood screening in blood donors
Fenglan YAO ; Rui WANG ; Jinghui HU ; Hongwei GE ; Chan LENG ; Yi ZHA ; Zifu ZHAO ; Zhengmin LIU
Chinese Journal of Blood Transfusion 2025;38(7):941-949
Objective: To validate the analytical performance, operational performance, and process control measures of a domestic fully automatic nucleic acid testing (NAT) system, thereby ensuring an efficient and orderly blood screening workflow. Methods: The concordance rate and sensitivity of WanTag-Vortex Plus system were verified using WHO standard reference panels of HIV-1, HCV and HBV, while precision was assessed using weak positive samples of HIV-1, HCV and HBV. As for its operational performance evaluation, cross-contamination resistance was assessed using strong positive samples, and throughput and stress testing were conducted using negative samples. Reagent stability was verified using weak positive samples, and inter-system performance consistency was assessed using verification panels. In addition, the process control measures were verified using the laboratory quality control demand scale. Results: 1) Verification of concordance rate: The detection results of negative and positive samples of HIV-1, HCV and HBV by WanTag-Vortex Plus system were all consistent with expectations, and the concordance rate was 100%. 2) Precision verification: the repeatability and intermediate precision were extremely high, and the coefficient of variation was less than 5%. 3) Verification of analytical sensitivity: The detection limit of 95% for standard strains of HIV-1, HCV and HBV by WanTag-Vortex Plus system in our laboratory was consistent with the analytical sensitivity provided by reagent manufacturers. 4) Verification of cross-contamination resistance: Five strong positive samples and 87 negative samples were placed according to the actual working conditions and equipment operation design, and the test results were consistent with expectations, with no cross-contamination in the testing system. 5) Throughput and stress testing: Each system completed the individual donor-nucleic acid amplification testing (ID-NAT) of 276 samples in three batches within 12 hours, and successfully completed the ID-NAT test of 828 samples in three consecutive days. 6) Verification of reagent stability: After extreme storage (unsealed storage for 1 week with 4 freeze-thaw cycles), the reagents maintained 100% detection rate in the weak positive samples of HIV-1, HCV, and HBV, showing no significant differences from the control group (Kappa=1). 7) Verification of inter-system performance consistency: The system has stable operation performance, and the performance comparison results across the four devices were consistent (Kappa=1). 8) Process control measures: WanTag-Vortex Plus system software accurately controlled the equipment operation process with strict quality control measures, and correctly interpreted and safely reported the test results. Conclusion: The analytical and operational performance of the WanTag-Vortex Plus system complies with manufacturer design standards and essential laboratory workflow requirements. Integrated with laboratory information system (LIS), the system's control software meets standard process control requirements, yet requires further improvement.
3.RRP15 Regulates Sensitivity of Hepatocellular Carcinoma to Sorafenib Through Ferroptosis
Saili ZHAO ; Zhangding WANG ; Fenglan WANG ; Lei WANG ; Bin ZHANG ; Xiaoping ZOU
Chinese Journal of Gastroenterology 2024;29(2):68-74
Background:Knockdown of ribosomal RNA processing 15 homolog(RRP15)inhibited the proliferation and growth of hepatocellular carcinoma(HCC),but its relationship with the sensitivity of HCC to sorafenib has not been reported.Aims:To elucidate the role of RRP15 in modulating the sensitivity of HCC to sorafenib and to unravel the underlying mechanisms.Methods:The constitutive expression of RRP15 in human HCC cell lines was assessed using real-time PCR and Western blotting,and then manipulated via infection with either RRP15 knockdown or overexpression lentivirus.The impact of RRP15 expression on sorafenib sensitivity of HCC cells was investigated by CCK-8 and colony formation assays.Changes in ferroptosis markers,including reactive oxygen species(ROS),Fe2+,lipid peroxide,reduced glutathione,etc in HCC cells were measured to determine the effect of RRP15 on ferroptosis.The combination of the ferroptosis inhibitor Ferrostatin-1 and RRP15 knockdown was used to verify the modulatory effect of RRP15 on sorafenib sensitivity and ferroptosis.Furthermore,xenograft tumor in nude mice was used to confirm the relationship between RRP15 and sorafenib sensitivity.Results:Sorafenib treatment induced RRP15 expression in HCC cells.The expression levels of RRP15 in HCC cells were negatively associated with the sensitivity to sorafenib.RRP15 knockdown enhanced the sorafenib sensitivity and sorafenib-induced ferroptosis in HCC cells,presenting as reduced cell viability,decreased colony formation ability,and increased intracellular ROS,Fe2+,and lipid peroxidation.Treatment with Ferrostatin-1 effectively compromised the increased ferroptosis and sorafenib sensitivity caused by RRP15 downregulation.Mechanistically,inactivation of p62-KEAP1-NRF2 pathway was involved in the RRP15 depletion-mediated ferroptosis and sorafenib sensitization in HCC cells.In in vivo study,RRP15 knockdown combined with sorafenib treatment notably inhibited the subcutaneous xenograft tumor growth in nude mice.Conclusions:This study demonstrates that inhibition of RRP15 significantly enhances the sensitivity of HCC to sorafenib,potentially through the promotion of ferroptosis.These findings may provide new strategies for improving the therapeutic response of HCC to sorafenib treatment.
4.Effects of Abiotic Elicitors MeJA and SA on Alkaloids Accumulation and Related Enzymes Metabolism inPinellia ternata Suspension Cell Cultures
Yongbo DUAN ; Fang LU ; Tingting CUI ; Fenglan ZHAO ; Jingtong TENG ; Wei SHENG ; Aimin ZHANG ; Jianping XUE
Chinese Journal of Information on Traditional Chinese Medicine 2017;24(1):87-90
Objective To study the effects of abiotic elicitors methyl jasmonate (MeJA) and salicylic acid (SA) on the alkaloids accumulation and related enzymes metabolism inPinellia ternata suspension cell cultures. Methods Using the leaf petioles-derived suspension cell cultures as the study object, the culture duration, concentrations of MeJA and SA were determined to get the optimal alkaloids accumulation, and the activities of metabolic enzymes IMP dehydrogenase and sAMP synthase were also measured.Results A 9-fold of dried biomass and a 3-fold of alkaloids accumulation were observed inP. ternata suspension cell cultures after culture for 21 d. Both MeJA and SA could significantly promote the accumulation of alkaloids inP. ternata suspension cells. 150 μmol/L MeJA enhanced alkaloids content (4.7 mg/gDW) by 3.6 folds in comparison with control group, whereas 50 μmol/L SA showed a 2.5-fold increase. Meanwhile, 100 μmol/L MeJA and 50 μmol/L SA promoted the increase in IMP dehydrogenase activity by 3.0 and 3.7 fold respectively, and 150 μmol/L MeJA and 100 μmol/L SA showed the increase by 2.6 and 4.4 fold respectively.Conclusion Proper adding exogenous MeJA and SA can promote the accumulation of alkaloids inPinellia ternata suspension cell cultures.
5.Effect of Clearing Heat and Resolving Phlegm on Blood Level of Neutrophil Elastase in AECOPD Rats
Jie LI ; Qi WANG ; Cuiling FENG ; Lishan ZHANG ; Zeji QIU ; Jun WU ; Hui LUO ; Fenglan DONG ; Fuding LIU ; Fucheng ZHAO ; Ying QI ; Guangwei XUE
China Pharmacist 2014;(5):722-724
Objective:To preliminarily study the effect of Louqin Zhisou decoctions with the mechanism of clearing heat and resol-ving phlegm on the blood level of neutrophil elastase ( NE) in the rat model of acute exacerbation of chronic obstructive pulmonary dis-ease ( AECOPD) . Methods:The rat model of AECOPD was established by passive cigarette smoking, intratracheal instillation of li-popolysacchricle ( LPY) and intranasal instillation of staphylococcus aureus. Totally 60 AECOPD rats were randomly divided into 3 groups, the model control group (n=20), ambrohexel group (n=20), and Louqin Zhisou decoctions group (n=20). NE was detec-ted by ELISA. Results:Compared with that before the treatment, NE in ambrohexel group and Louqin Zhisou decoctions group was de-creased significantly(P<0. 01). Conclusion:Clearing heat and resolving phlegm method probably can decrease NE by reducing air-way mucus hypersecretion and obstruction in AECOPD rats.
6.Expression of potassium ion channel mRNA in myocardial tissue of mice with low selenium
Meina SUN ; Fenglan LI ; Handong ZHAO ; Hui LI
Chinese Journal of Endemiology 2014;33(1):46-48
Objective To examine the expression of potassium channel mRNA in myocardial tissue of mice with low selenium.Methods Forty C57BL/6 mice were randomly divided into four groups according to body weight(18-22 g),10 mice in each group,half male and half female.The low-selenium treatment groups were fed with low-selenium diet(selenium content was 0.004 mg/kg) for 4,12 and 24 weeks,respectively,and the control group was fed with normal diet(selenium content was 0.256 mg/kg).The mice were killed by cutting neck method,hearts were taken out and RNA was extracted by Trizol method.The expressions of potassium ion channel genes (KCNA4,KCND2,KCND3,KCNE1,KCNE2,KCNJ2,KCNJ12 and KCNQ1) at the mRNA level in heart were determined using real-time polymerase chain reaction.Results In low-selenium 4 weeks group,the mRNA expressions of KCNA4 gene(25.3 ± 0.09) and KCND2 gene(4.85 ± 0.05) were higher than that of the control group (1.00 ± 0.00,1.00 ± 0.00,all P < 0.05); in low-selenium 24 weeks group,the mRNA expression of KCNJ12 gene (22.7 ± 0.10) was higher than that of the control group(1.00 ± 0.00,P < 0.05); the mRNA expressions of KCND3,KCNE1,KCNE2,KCNJ2,KCNQ1 genes were compared with the corresponding control groups,the differences were not statistically significant(all P > 0.05).Conclusions Low selenium can affect the mRNA expression of mouse cardiac potassium ion channel genes.
7.The relationships among organization climate, psychological empowerment and job embeddedness
Yixin ZHAO ; Yang ZHAO ; Jiali FU ; Xiaoyun CHEN ; Wenfang WU ; Fenglan LOU
Chinese Journal of Practical Nursing 2014;30(13):29-32
Objective To explore the relationships among organization climate,psychological empowerment and job embeddedness by using path analysis.Methods A total of 514 clinical nurses from 26 departments in 3 hospitals were recruited by convenience cluster sampling method and investigated with demography questionnaire,Nurses Organizational Climate Scale,Psychological Empowerment Scale and Job Embeddedness Scale.Results The mean scores of organization climate,psychological empowerment and job embeddedness was (3.01±0.46),(3.27±0.44),(3.08±0.39).The predictors of job embeddedness of nurses were resources support,work experience,human resources management,marital status,job title,management support,quality management,and self-egicacy,explained 48.9% of its variance; organization climate,psychological empowerment had direct positive influence on nurses' job embeddedness,explained 33.3% of its variance.Conclusions Improvement and maintenance of sound organizational climate,increase the sense of psychological empowerment of nurses,are effective ways and means to increase the degree of job embedding of nurses.
8.Relationship between expression of selenocysteine synthase and endothelial cell line ECV-304 injury induced by hydrogen peroxide
Handong ZHAO ; Meina SUN ; Fenglan LI ; Hui LI
Chinese Journal of Endemiology 2014;(4):383-386,387
Objective To observe the influence of selenocysteine synthase(SEPSECS) on injury of human umbilical vein endothelial cell line EVC-304 induced by hydrogen peroxide (H2O2). Methods Transfection was conducted to transfect EVC-304 which was maintained in vitro. The cells were divided into four groups: control group, SEPSECS over-expression group, empty vector group and SEPSECS silenced expression group, then Real-time PCR and Western blotting were performed to detected SEPSECS mRNA and protein expression , respectively. Flow cytometry(FCM) was performed to detect cell cycle. Different concentrations of H2O2, which including 0, 200, 400, 600, 800, 1 000 μmol/L, were used to treat EVC-304 . Then malonaldehyde (MDA), superoxide dismutase(SOD) secreted by the cells which were treated with H2O2 for 6 h, were checked by MDA or SOD kit. Results The SEPSECS mRNA expressions of control, SEPSECS silenced expression, empty vector and SEPSECS over-expression groups were 1.03 ± 0.24, 0.43 ± 0.11, 0.98 ± 0.27 and 1.61 ± 0.13, respectively. The protein expressions of control, SEPSECS silenced expression, empty vector and SEPSECS over-expression groups were 1.00 ± 0.26, 0.51 ± 0.10, 1.12 ± 0.38 and 1.51 ± 0.20, respectively. There was a significant difference between control and SEPSECS silenced expression groups (all P<0.05), at the same time , this phenomenon was also observed between empty vector and SEPSECS over-expression groups (all P<0.05). The level of MDA was increased along with the H2O2 concentration. Besides, cell cycle was also tested, however, significant difference was not observed(all P > 0.05). Meanwhile, MDA of SEPSECS silenced expression groups[(15.8 ± 0.5),(19.6 ± 1.5)μmol/L] were significantly higher than control groups[(12.4 ± 0.1),(17.1 ± 0.5)μmol/L, all P < 0.05], on the other hand, MDA of SEPSECS over-expression groups[(10.8 ± 0.4),(14.2 ± 1.1)μmol/L] were lower than empty vector groups [(12.7 ± 0.7),(16.2 ± 1.1)μmol/L, all P < 0.05], when the H2O2 concentration was 800 or 1 000μmol/L. The level of SOD was decreased along with the H2O2 concentration. SOD of SEPSECS silenced expression groups[(7.7 ± 0.4),(2.4 ± 0.3)μmol/L] were lower than control groups[(10.0 ± 1.0),(6.0 ± 0.6)μmol/L, all P < 0.05], on the contrary, SOD of SEPSECS over-expression groups[(11.3 ± 0.6),(12.7 ± 1.6)μmol/L] were higher than empty vector groups[(9.2 ± 0.6),(6.7 ± 0.2)μmol/L, all P < 0.05], when the H2O2 concentration was 800 or 1 000μmol/L. Conclusion Expression of SEPSECS has a significant protective role on damaged EVC-304 which was induced by H2O2.
9.Preparation and clinical application of a nose prosthesis by means of selective laser simtering and wax powder
Fenglan LI ; Yimin ZHAO ; Guofeng WU
Chinese Journal of Medical Aesthetics and Cosmetology 2009;15(6):384-386
Objective To explore the clinical application of reconstructing silicone elastomer nose prosthesis by means of selected laser sintering and wax powder PCPI. Methods Laser scanning was used to get the 3-D data of a nose model. Surfacere 10.0 etc softwares was used to reconstruct the nose by mirroring the digitalized model of absent nose. Selective laser sintering and wax powder was chosen to fabricate a wax nose model and the nose prosthesis made by silicone clastomer. Results Perfect silicone clastomer nose prosthesis was made for 2 patients. Conclusion This study suggests that the wax nose model and the new wax powder can meet the requirement of clinical expectation for maxillofacial prosthesis.
10.Preparation and properties of 5-fluorouracil-loaded chitosan microspheres for the intranasal administration
Aiping ZHENG ; Shaoyun YU ; Ying ZHAO ; Fenglan WU
Journal of Peking University(Health Sciences) 2004;0(03):-
Objective: To study the preparation technique and release characteristic of 5- fluorouracil-loaded chitosan microspheres for the intranasal administration. Methods:Using the liquid paraffin as the oil phase,and span-80 as the emuifier; 5- fluorouracil-loaded chitosan microspheres were achieved by emulsion-chemical crosslink technique. The orthogonal experimental design was applied to optimize the preparation procedure. Dynamic dialysis method was used to determine the releasing characteristic of microspheres in vitro and it influencing fators.Swelling behavior was expressed by swelling ratio.The degree of mucoadhesion was investigated by determining the mucociliary transport rate(MTR) of the microparticle across a frog palate. Results: Microspheres with a good shape and narrow size distribution were prepared. The average diameter was (43?4) ?m. The drug loading was 38.5%? 1.0%. The entrapment efficiency was 79.0%?1.8%.The drug release profile in vitro could be described by Higuichi eqution Q=0.1035t 1/2 +0.0284 (r=0.9965). Chitosan had good mucoadhesive property and caused a siginificant reduction in MTR(P

Result Analysis
Print
Save
E-mail