1.The role of IDO/TTS mediated tryptophan metabolic pathway in patients with immune thrombocytopenia
Qiujie CHEN ; Yu ZENG ; Yunlong GAO ; Yan MA ; Xinzhong WU
The Journal of Practical Medicine 2014;(21):3441-3444
Objective To discuss the role of indoleamine 2, 3-dioxygenas e (IDO) and tryptophanyl-tRNA synthetase (TTS) mediated tryptophan catabolism in immune thrombocytopenia (ITP) patients treated with high doses of dexamethasone through the expressions of IDO and TTS in T cells , and the concentrations of plasma kynurenine and tryptophan. Methods 20 newly diagnosed or relapse ITP patients were treated with 40 mg/d × 4 d dose of dexamethasone. The heparin anticoagulant blood samples were obtained before treatment and the 5th day after treatment. 20 healthy subjects were selected as the control group. The IDO and TTS expressions in CD4+and CD8+ T cells were analyzed by flow cytometry. The concentrations of plasma kynurenine and tryptophan were detected by liquid-mass spectrometry system. Results Compared with healthy controls group, the plasma tryptophan and kynurenine concentration and the ratio of Kyn/Trp were significantly elevated in ITP patients (P <0.05); the IDO expressions of CD4+ and CD8+ T cells in ITP patients were significantly lower than those in healthy controls (P < 0.05), but the TTS expressions were significantly higher (P < 0.05). The concentration of tryptophan in effective group was significantly lower than before treatment (P < 0.05), in contrast, the kynurenine concentration and the ratio of Kyn/Trp were significantly higher than before (P < 0.05). The expression of IDO in effective group were significantly higher than that before treatment (P < 0.05), conversely, the expression of TTS in effective group were significantly decreased (P < 0.05). No significant difference can be found in ineffective group. Conclusion IDO/TTS-mediated tryptophan catabolism pathway could indicate the onset of ITP. The sensitivity of ITP patients with high dose of dexamethasone treatment can be observed through the level of IDO and TTS.
2.Effects of miR-145 on the proliferation, apoptosis and cell cycle of a human keratinocyte cell line HaCaT
Jingrong LI ; Jianqin WANG ; Ruihua FANG ; Renshan ZENG ; Jinxue MO ; Yunlong GUO ; Shuqing JIA
Chinese Journal of Dermatology 2015;48(6):416-420
Objective To investigate the regulatory effects of miR-145 on the proliferation,cell cycle and apoptosis of a human keratinocyte cell line HaCaT.Methods miR-145 mimics and negative control (NC) mimics were chemically synthesized and then transiently transfected into HaCaT cells respectively.After additional culture for different durations,real-time PCR was performed to determine the expression level of miR-145,MTS assay to estimate cell proliferation,and flow cytometry to detect cell apoptosis and cycle.Luciferase assay,real-time PCR and Western blot were conducted to determine whether NRAS was the target gene of miR-145.Results The miR-145 expression level in miR-145 mimic-transfected cells increased by 85.00 ± 1.21 folds compared with NC mimic-transfected cells (t =115.90,P < 0.0001).The transfection with miR-145 mimics significantly inhibited the proliferation of HaCaT cells (F =8.76,P =0.008),and the inhibitory effect significantly varied with the duration (24-96 hours) of culture after transfection,with no interaction effect between the transfection with miR-145 mimics and culture duation (F =1.21,P =0.18).Compared with NC mimic-transfected cells,those transfected with miR-145 mimics showed a significant increase in the proportion of early apoptotic cells (18.9% ± 4.1% vs.4.3% ± 1.2%,t =7.126,P < 0.01),late apoptotic cells (9.3% ± 2.3% vs.3.6% ± 1.6%,t =12.38,P < 0.01),G1-phase cells (85.83% ± 5.2% vs.62.08% ± 6.23%,t =11.78,P =0.007),but a significant decrease in the percentage of G2-phase cells (6.26% ± 1.2% vs.19.36% ± 3.45%,t =7.610,P =0.017) and S-phase cells (7.91% ± 1.3% vs.18.56% ± 5.23%,t =7.230,P=0.019).As luciferase assay showed,luciferase activity was significantly lower in HaCaT cells cotransfected with miR-145 mimics and a recombinant luciferase reporter vector psi-CHECK2-NRAS-wild carrying the wild-type 3'UTR of NRAS than in those cotransfected with NC mimics and the vector psi-CHECK2-NRAS-wild (t =11.09,P =0.008),but similar between cells cotransfected with miR-145 mimics and a recombinant luciferase reporter vector psi-CHECK2-NRAS-mut carrying the mutant-type 3'UTR of NRAS and those cotransfected with NC mimics and the vector psi-CHECK2-NRAS-mut (P > 0.05).Real-time PCR and Western blot revealed that the overexpression of miR-145 mimics had no significant effect on NRAS mRNA expression (P > 0.05),but significantly inhibited NRAS protein expression (1.52 ± 0.07 vs.0.20 ± 0.02,t =28.43,P< 0.01).Conclusion miR-145 might inhibit proliferation and promote apoptosis of HaCaT cells by influencing cell cycle via NRAS.
3.Study on rejoin detection mode for unpaid blood donors with HBsAgor anti-HCV positive reactivity
Jinfeng ZENG ; Yunlong CHEN ; Xin ZHENG ; Heng LIU ; Linfeng WU ; Dandan DU
International Journal of Laboratory Medicine 2014;(23):3207-3209
Objective To investigate the detection mode for those unpaid blood donors screening as positive HBsAg+ and/or positive anti-HCV could be permitted to recall again for re-detection under the regulation condition in order to determine whether or not regain their qualifications of donating blood and rejoin the blood donation again.Methods The unpaid blood donors preliminari-ly screening as positive HBsAg and/or positive anti-HCV in Shenzhen from Otcober 2007 to December 2013 and conforming to the rules for recalling to re-detection formulated by our center were analyzed and researched for conducting the feasibility discussion on the rejoin mode of unpaid blood donors.Results A total of 415 759 case-times of blood donation were conducted during 2007 ~2013.Among them,2 506 cases(0.60%)and 1 357 cases(0.33%)were screened as positive HBsAg or positive anti-HCV,respec-tively.The recall process of rejoin re-detection was initiated in 59 positive HBsAg donors and 16 positive anti-HCV donors with many times of blood donation.But only 31 positive HBsAg donors and 9 positive anti-HCV donors successfully completed the detec-tion items of re-detection process.Among them 29 positive HBsAg donor regained the qualifications of donating blood and 2 cases were shielded for the blood donation qualification due to unqualification in the following detection.All of the 9 recalled donators with positive anti-HCV regained the blood donation qualification.Conclusion Under present detection mode,the detection tech-nique of blood screening is hard to avoid the occurrence of false positive results caused by the reagents,instruments and personnel operating.In order to protect the donation qualifications of the unpaid blood donators,a set of scientific,reasonable and practical re-detection mode for rejoin of the blood donators should be established for protecting the limited blood donation resource.
4.Induction of osteogenic differentiation of human renal fibroblasts in vitro
Yijie YAN ; Chengyang LI ; Yaoliang DENG ; Guohua ZENG ; Zhiwei TAO ; Yunlong LIU ; Chun SUN ; Yang WANG
Chinese Journal of Pathophysiology 2015;(12):2259-2264
AIM:To investigate the effects of osteogenic induction media and the medias containing different concentration of calcium on the induction of osteogenic differentiation of human renal fibroblasts in vitro.METHODS: Culturedhuman renal fibroblasts were divided into 5 groups in this experiment: osteogenic induction group (osteogenic inductionmedia), CaⅠgroup (0.5 mmol/L Ca2 + media), CaⅡgroup (1.5 mmol/L Ca2 + media), Ca Ⅲ group (2.5 mmol/LCa2 + media) and control group (PBS).The cell activity in each groups was measured by MTT assay .At 9th day, the cellcalcium Alizarin red S staining and alkaline phosphatase (ALP) Gomori calcium cobalt staining were performed respectivelyto observe the formation of calcium nidus and the expression of ALP .In addition, the expression of Runt-related transcriptionfactor 2 (Runx2) at mRNA and protein levels was determined by real -time PCR and Western blot, respectively.RE- SULTS: The remarkable positive signs which represented the formation of calcium nidus and the deposit of calcium objectsin all experiment groups were observed .The mRNA and protein expression of Runx2 in osteogenic induction group increasedin accordance with the induction time .Compared with control group, the mRNA and protein expression of Runx2 inthe CaⅠ ~Ⅲ groups increased gradually in a calcium concentration dependent manner at the 9th induction day.CON- CLUSION: Human renal interstitial fibroblasts show the potential activity in osteogenic differentiation induced by osteogen -ic induction media or high level calcium in vitro, which may be account for the cytological formation of the Randall ’splaque in the kidney.
5.Determination of Moroxydine Residue in Tomatoes Using CdTe Quantum Dots as Fluorescence Probes
Xiaoyun YUAN ; Huiyan KUANG ; Lei FENG ; Haowen HUANG ; Chunran TANG ; Yunlong ZENG
Chinese Journal of Analytical Chemistry 2014;(7):1057-1060
A novel method for the detection of moroxydine ( ABOB) residue in tomatoes was developed based on the enhancement of the fluorescence intensity of CdTe quantum dots( QDs) in the presence of ABOB. The factors influencing the performance of the QDs fluorescent probes were investigated, and the optimum conditions were determined:the concentration of mercaptoacidic acid ( TGA) capped-CdTe quantum dot was 1×10-4 mol/L, and the reaction time was 20 min at pH=5. 6. Under the optimum conditions, the relative fluorescence intensity increases linearly proportional to the ABOB concentration in the range of 1. 0×10-12-5. 0×10-10 mol/L with a limit of detection of 5. 2×10-13 mol/L, R=0. 9981, the recovery was 97%-106%, without obvious interference on the determinations of moroxydine from the common coexisting ions, antibiotics, and vitamins. The proposed method has been successfully applied in the detection of trace moroxydine hydrochloride residue in tomatoes.
6.Improvement of synthesis method of 6-methoxyisatin
Hongyu WANG ; Yunlong SONG ; Ling ZHANG ; Qiang ZENG ; Ju ZHU ; Hui ZHANG
Journal of Pharmaceutical Practice 2015;(2):127-130
Objective To improve the synthetic condition of 6‐methoxyisatin .Methods The starting material 3‐me‐thoxyaniline was firstly converted into the intermediate 1‐oximino‐N‐ (3‐methoxyphenyl) acetamide through Sandmeyer reac‐tion ,then 6‐methoxyisatin was conducted with methanesulfonic acid as catalyst in the following ring closure reaction .Results In the second step of the cyclization reaction ,methanesulfonic acid was used to replace the concentrated sulfuric acid which is widely used in literature .The temperature was 80℃ ,the reaction time was 30 min ,and the yield was 81.24% .Conclusion This study provided a synthesis process of 6‐methoxyisatin with simple operation ,mild reaction condition and high yield ,which is suitable for large scale preparation of the compound .
7.Construction of risk prediction model for predicting death or readmission in acute heart failure patients during vulnerable phase based on machine learning
Jing ZENG ; Xiaolong HE ; Huajuan HU ; Xiaoyu LUO ; Zhinian GUO ; Yunlong CHEN ; Min WANG ; Jiang WANG
Journal of Army Medical University 2024;46(7):738-745
Objective To construct risk prediction models of death or readmission in patients with acute heart failure(AHF)during the vulnerable phase based on machine learning algorithms and screen the optimal model.Methods A total of 651 AHF patients with admitted to Department of Cardiology of the Second Affiliated Hospital of Army Medical University from October 2019 to July 2021 were included.The clinical data consisting of admission vital signs,comorbidities and laboratory results were collected from electronic medical records.The composite endpoint was defined as all-cause death or readmission for worsening heart failure within 3 months after discharge.The patients were divided into a training set(521 patients)and a test set(130 patients)in a ratio of 8:2 through the simple random sampling.Six machine learning models were developed,including logistic regression(LR),random forest(RF),decision tree(DT),light gradient boosting machine(LGBM),extreme gradient boosting(XGBoost)and neural networks(NN).Receiver operating characteristic(ROC)curve and decision curve analysis(DCA)were used to evaluate the predictive performance and clinical benefit of the models.Shapley additive explanation(SHAP)was used to explain and evaluate the effect of different clinical characteristics on the models.Results A total of 651 AHF patients were included,of whom 203 patients(31.2%)died or were readmitted during the vulnerable phase.ROC curve analysis showed that the AUC values of the LR,RF,DT,LGBM,XGBoost and NN model were 0.707,0.756,0.616,0.677,0.768 and 0.681,respectively.The XGBoost model had the highest AUC value.DCA showed that the XGBoost model exhibited greater clinical net benefit compared with other models,with the best predictive performance.SHAP algorithm analysis showed that the clinical features that had the greatest impact on the output of the model were serum uric acid,D-dimer,mean arterial pressure,B-type natriuretic peptide,left atrial diameter,body mass index,and New York Heart Association(NYHA)classification.Conclusion The XGBoost model has the best predictive performance in predicting the risk of death or readmission of AHF patients during the vulnerable phase.
8.Results analysis of unexpected antibody screening for blood donors and cost comparison of two detection modes
Liqin HUANG ; Lifeng WU ; Tong LI ; Ran LI ; Heng LIU ; Dandan DU ; Yunlong CHEN ; Xuezheng ZENG ; Jinfeng ZENG
Chinese Journal of Blood Transfusion 2022;35(3):324-327
【Objective】 To establish a routine screening method for unexpected antibodies of blood donors, analyze the results of centralized screening for unexpected antibody of blood donors in the blood center, and compare the cost of centralized and decentralized screening modes. 【Methods】 A total of 35 591 blood donors were screened for unexpected antibodies from March 31, 2021 to July 31, 2021, using microcolumn gel method. Unexpected antibody screening reactive samples were further confirmed by the Transfusion Research Institute of Shenzhen Blood Center, and the demographic characteristics were further determined through the analysis of unexpected antibody positive population. The direct cost and indirect cost of centralized and decentralized unexpected antibody screening mode were compared. 【Results】 Forty unexpected antibody positive samples were confirmed in Shenzhen, with the positive rate at 0.11%(40/35 591), among which MNS, Rh and Lewis system accounted for 35% (14/40), 32.5% (13/40) and 17.5% (7/40), respectively. Males and females accounted for 45% (18/40) and 55% (22/40), respectively (P<0.01). No significant difference was noticed by age and repeated-donor or not (P>0.05). Unexpected antibody screening in a centralized way saved about 1.16 million yuan per year. 【Conclusion】 It is necessary to carry out unexpected antibody screening for all blood donors, and centralized screening is more economical than decentralized screening.
9.Tactic movement of microalgae and its application in targeted transport: a review.
Yuanyuan LIU ; Weiyang ZENG ; Ru CHEN ; Yunlong GE ; Lihan ZI ; Jun YANG ; Fantao KONG
Chinese Journal of Biotechnology 2022;38(2):578-591
Microalgae are a group of photosynthetic microorganisms, which have the general characteristics of plants such as photosynthesis, and some species have the ability of movement which resembles animals. Recently, it was reported that microalgae cells can be engineered to precisely deliver medicine-particles and other goods in microfluidic chips. These studies showed great application potential in biomedical treatment and pharmacodynamic analysis, which have become one of the current research hotspots. However, these developments have been rarely reviewed. Here, we summarized the advances in manageable movement exemplified by a model microalgae Chlamydomonas reinhardtii based on its characteristics of chemotaxis, phototaxis, and magnetotaxis. The bottlenecks and prospects in the application of microalgae-based tactic movement were also discussed. This review might be useful for rational design and modification of microalgal manageable movement to achieve targeted transport in medical and other fields.
Chlamydomonas reinhardtii
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Microalgae
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Microfluidics
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Photosynthesis