2.Preparation of targeting CPI-444-loaded nanoparticles and investigation of its effects on T cell activity and anti-tumor response
CHEN Mingshui ; LI Jieyu ; WANG Ling ; ZHOU Zhifeng ; ZHANG Linteng
Chinese Journal of Cancer Biotherapy 2024;31(8):777-785
[摘 要] 目的:制备并表征包载CPI-444并偶联CD8抗体的纳米微粒(CNP/αCD8),探讨其对CD8+ T细胞活化、增殖和抗肿瘤作用的影响。方法: 采用复乳溶剂蒸发法和EDC/NHS法制备包载腺苷受体A2A(A2AR)特异性拮抗剂CPI-444(C)或香豆素6(C6)荧光素的纳米微粒并分别在其表面偶联CD8抗体,制得CNP/αCD8和C6NP/αCD8。扫描电镜和NanoPlus粒度测定仪表征纳米微粒形态和粒径,液相色谱与串联质谱联用(LC-MS/MS)法和离心法测定纳米微粒的载药量和药物释放情况,荧光显微镜和流式细胞仪检测CD8+ T细胞内化C6NP/αCD8的情况,流式细胞仪、ELISA和LDH法检测CNP/αCD8对CD8+ T细胞增殖、活化、细胞毒活性和杀瘤能力的影响。结果: CNP/αCD8纳米微粒为圆形、粒径约150 nm,能有效包载CPI-444和偶联CD8抗体,药物包封率和CD8抗体偶联效率分别约为60%和53.4%;CNP/αCD8纳米微粒具有良好稳定性,能被CD8+ T细胞内化,抑制A2AR分子表达。生物学功能实验显示,CNP/αCD8增强CD8+ T细胞的增殖能力、促进T细胞活化、分泌细胞因子及产生颗粒酶B和穿孔素,并增强CD8+ T细胞杀伤肿瘤细胞的能力。结论: CNP/αCD8纳米微粒能显著增强CD8+ T细胞免疫效应功能,其增强CD8+ T细胞功能可能是通过抑制A2AR分子的表达起作用。
3.Relationship between Iron Metabolic Parameters and Platelet Counts in Blood Donors.
Wen-Juan ZHONG ; Qiu-Fang ZHANG ; Cheng-Yong HUANG ; Ying-Chun CHEN ; Ye-Ping ZHOU ; Jin-Ying CHEN ; Jia ZENG
Journal of Experimental Hematology 2023;31(5):1481-1485
OBJECTIVE:
To investigate the correlation of iron metabolic parameters with platelet counts in blood donors.
METHODS:
A total of 400 blood donors who met requirements of apheresis platelet donation were collected, and their hematological parameters were analyzed. The donors were divided into low ferritin group and normal group, the differences of hematological parameters between the two groups were compared, and the correlation of iron metabolic parameters and routine hematology parameters with platelet counts were analyzed.
RESULTS:
Whether male or female, low ferritin group had higher platelet counts than normal group (P < 0.01). Among the iron metabolic parameters, the platelet counts was negatively correlated with serum ferritin (SF), serum iron (SI), and transferrin saturation (TSAT) (r =-0.162, r =-0.153, r =-0.256), and positively correlated with total iron binding capacity (TIBC) and unsaturated iron binding capacity (UIBC) (r =0.219, r =0.294) in female blood donors. Platelet counts was also negatively correlated with SF, SI and TSAT (r =-0.188, r =-0.148, r =-0.224) and positively correlated with UIBC (r =0.220) in male blood donors. Among the routine hematology parameters, platelet counts was negatively correlated with mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH), mean corpuscular hemoglobin concentration (MCHC), and reticulocyte hemoglobin equivalent (Ret-He) in female blood donors (r =-0.236, r =-0.267, r =-0.213, r =-0.284). Platelet counts was also negatively correlated with MCH, MCHC and Ret-He in male blood donors (r =-0.184, r =-0.221, r =-0.209).
CONCLUSION
In blood donors with low C-reactive protein level, the lower the iron store capacity, the lower the iron utilization, and the platelet counts tends to rise.
Male
;
Humans
;
Female
;
Iron/metabolism*
;
Blood Donors
;
Platelet Count
;
Anemia, Iron-Deficiency
;
Hemoglobins
;
Ferritins
4.Analysis of Iron Stores in the Plateletpheresis Donors.
Wen-Juan ZHONG ; Ben-Chun REN ; Ye-Ping ZHOU ; Xiao-Mei LIN ; Ming WANG
Journal of Experimental Hematology 2019;27(3):925-929
OBJECTIVE:
To understand the iron stores of the plateletpheresis donors, so as to provide some new experimental data for further exploration and more perfect health examination criteria of the plateletpheresis donors.
METHODS:
A total of 297 plateletheresis donors conformed to standard in October 2018 were selected by the cross sectional study. The related factors affecting iron stores were analyzed; the effect of plateletpheresis times of donation on the levels of the hemoglobin(Hb) and serum ferritin(SF) as well as the iron deficency rate in the blood donors was also analyzed; the iron stores in the blood donors was evaluated.
RESULTS:
The SF level in plateletpheresis donors negatively correlated with annual plateletphersis times of donation(r=-0.416, P<0.001); The SF level decreased with the increase of annual times of donation(P<0.05); The iron deficiency rate in plateletpheresis donors showed the increase trend with the increase of annual times of donation. The iron deficiency rate in male and femal with 18-23 times of donation was 12.5%(8/64) and 40%(6/15) respectively.
CONCLUSION
The blood center should reduce recruitment frequency and increase the testing of SF for regularly plateletpheresis donors.
Blood Donors
;
Cross-Sectional Studies
;
Ferritins
;
Hemoglobins
;
Humans
;
Iron
;
Male
;
Plateletpheresis
5.Treatment of erectile dysfunction in patients with spinal cord injury.
National Journal of Andrology 2017;23(2):99-102
The majority of men with spinal cord injury (SCI) suffer from different types and degrees of erectile dysfunction (ED). The impact of SCL on erectile function depends on the severity and location of the injury. Some patients can restore the residual sexual potential in the period of SCI recovery. Treatments of ED in men with SCI include psychotherapy, medication of oral PDE5 inhibitors, intracavernous injection of vasoactive drugs, transurethral medication, application of the vacuum erectile device and penile ring, penile prostheses, sacral neuromodulation, etc., among which PDE5 inhibitors remain the first option and usually give good clinical results for those with higher SCI. However, intracavernous injection of vasoactive drugs and combined treatment are preferable for those with lower SCI.
Erectile Dysfunction
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etiology
;
therapy
;
Humans
;
Male
;
Penile Prosthesis
;
Phosphodiesterase 5 Inhibitors
;
therapeutic use
;
Psychotherapy
;
Spinal Cord Injuries
;
complications
6.Application of TMA Technology in Donors' HBV-DNA Detection.
Ben-Chun REN ; Xiao-Zhen ZHOU ; Xiao-Fu ZHUO
Journal of Experimental Hematology 2019;27(6):1998-2002
OBJECTIVE:
To understand the infection of hepatitis B virus(HBV) in blood donors, and to evaluate the effectiveness and necessity of TMA technology for HBV-DNA screening in blood donors.
METHODS:
Using the ELISA/NAT model, routine serology test and NAT were performed in the 169 160 donors,including voluntary blood donors and some of donors returned to donor team. For some donors with test positive NAT, nucleic acid identification test was performed. And the HBsAg neutralized and confirmed assay would conduct in blood donors with unilateral HBsAg positive and HBV-DNA negative result.
RESULTS:
Among 169 160 donation cases-times, the donors of bilateral positive of HBsAg detection was 803, accounted for 0.476%; donors of unilateral positive was 243, accounted for 0.144%. For 40 specimens with HBV-DNA negative, unilateral HBsAg positive, the neutralization and confirmed assay was performed.In result, only 4 specimens were confirmed to be HBsAg positive, the confirmed positive rate was 10%. Among detected 1003 specimens with HBV-DNA positive specimens, both HBsAg and HBV-DNA positive were 739, the consistency rate between 2 kinds of detection was 73.7%. The comparision of positive rate detected by using 3 kinds of reagents showed that there were statistical differences (P<0.05); moreover, there were statistical difference in positive rate detected by using Murex reagent and In Tec reagent (P<0.0125). The comparison of detected rate of HBsAg and HBV-DNA during March 2016-February 2017 showed no statistical difference (P>0.05). Among 60 blood donors with HBsAg and HBV-DNA who has retured to the donor team, 1 donor presented the transformation of HBsAg from negative to positive, suggesting the HBV infection of window period, HBsAg of the other 59 was negative. The detection of HBV-DNA showed that the HBV-DNA in 28 donors was negative, and the HBV-DNA in 31 donors was positive, 1 donor showed HBV-DNA was uncertain.
CONCLUSION
The routine TMA technology combined with ELISA HBsAg can effectively shorten the window period for detection of HBV infection, effectively detect the occult HBV infection, and reduce the potential risk of hepatitis B spread due to blood transfusion.
Blood Donors
;
DNA, Viral
;
Hepatitis B
;
Hepatitis B Surface Antigens
;
Hepatitis B virus
;
genetics
;
Humans
7.Pyramiding of 3-resistant-gene to improve rice blast resistance of a restorer line, Fuhui 673.
Zhiwei CHEN ; Huazhong GUAN ; Xiaofang WANG ; Ruixia DONG ; Chenghai ZHUO ; Damei MAO ; Runsen PAN ; Yuanchang ZHOU ; Weiren WU
Chinese Journal of Biotechnology 2019;35(5):837-846
To improve the blast resistance of elite rice restorer line Fuhui 673, 3 blast resistance genes Pi-1, Pi-9 and Pi-kh were introduced into Fuhui 673 from a good-quality restorer line Jinhui 1059 through 3 successive backcrosses followed by one selfing using the technique of marker-assisted selection. Ten near-isogenic lines (NILs) of Fuhui 673 carrying the 3 introduced resistance genes were created. Genotype analysis using 68 SSR markers evenly distributed in the genome indicated that 92.96%-98.59% of the NILs' genetic background had been recovered to Fuhui 673. Both indoor and field resistance tests indicated that the NILs and their hybrids with sterile line Yixiang A were all resistant to rice blast, with resistance levels significantly higher than those of controls Fuhui 673 and hybrid Yiyou 673 (Yixiang A Fuhui 673). In addition, among the 10 hybrids between the NILs and Yixiang A, 2 showed significantly higher yield than and 4 displayed similar yield to that of control Yiyou 673, suggesting that most of the NILs retained the elite characteristics of Fuhui 673. Two new hybrid rice cultivars Liangyou 7283 and Jintaiyou 683 from NIL Line 9 showed high yield, good resistance to blast and moderate growth period in regional trial, suggesting that the NIL Line 9 has a good prospect for application.
Breeding
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Disease Resistance
;
genetics
;
Genes, Plant
;
genetics
;
Oryza
;
genetics
8.A Case-Control Study on Receptor Gene Polymorphism and Risk Suffering from Adult Acute Leukemia in Fujian Area.
Feng ZHANG ; Hai-Ying FU ; Hua-Rong ZHOU ; Rong CHEN ; Jian-Zhen SHEN
Journal of Experimental Hematology 2021;29(1):1-8
OBJECTIVE:
To investigate the correlation of receptor gene (P2X7, VDR and SLC19A1) polymorphisms with risk suffering from acute leukemia (AL) in Fujian area.
METHODS:
Ninety-three cases of newly diagnosed AL as AL group and 90 persons not suffered from hematologic and other tumors as control group were selected and used for comparative analysis of receptor gene polymorphisms and risk suffering from AL between case and control groups. The bone marrow and peripheral blood were collected, from which the DNA was extracted. The PCR-RFLP was used to detect 8 SNP sites (P2X7: rs208294, rs2230911, rs3751143; VDR: rs2228570, rs7975232; SLC194A1: rs1051266, rs1131596, rs3788200) of receptor genes related with the environment response, and the genotypes analysis was used to the correlation of receptor gene polymorphisms with risk suffering from adult AL.
RESULTS:
The unvariate logistic analysis showed that as compared with control group, P2X7 rs208294 T>C mutation and rs3751143 A>C mutation in codominant model, dominant model and over-dominant model were higher in case group, moreover the differences were statistically significant (P<0.01, P<0.05 and P<0.05, respectively), which suggested that they could reduce the risk suffering from AL. The recessive inheritance model showed that SLC1941 rs1131596 G>A mutation could increase the risk suffering from AL (P<0.05). The stepwise multivariate logistic regression analysis showed that there was still statistically significant difference in P2X7 rs208294 mutation between case group and control group (P<0.05), moreover, the heterozygous mutation (CT) could decrease the risk suffering from AL, showing the better protective effect, compared with homozygous mutation(CC).
CONCLUSION
The P2X7 rs208294 T>C mutation is one of protective factors against adult acute leukemia.
Adult
;
Case-Control Studies
;
Gene Frequency
;
Genetic Predisposition to Disease
;
Genotype
;
Homozygote
;
Humans
;
Leukemia, Myeloid, Acute
;
Polymorphism, Genetic
;
Polymorphism, Single Nucleotide
;
Receptors, Purinergic P2X7
9. Preparation of silymarin-SBE-β-CD inclusion complex
Chinese Traditional and Herbal Drugs 2019;50(17):4100-4107
Objective: The inclusion compound of silymarin-SBE-β-CD was prepared. Methods: Phase solubility method was used to screen the inclusion materials of cyclodextrin and determine the type of inclusion. The technological conditions of silymarin inclusion complex were optimized by orthogonal design. The solubility of inclusion was determined and its structure was characterized by means of microscope, infrared spectrum analysis and X-ray diffraction analysis. Results: Due to its higher solubilization effect on silymarin, SBE-β-CD was determined as inclusion material. The ratio of drug to cyclodextrin was coated in the mode of 1:n. The process optimized by orthogonal design was as follow: molar ratio of silymarin to SBE-β-CD of 1:8, inclusion temperature of 60 ℃, and inclusion time of 3 h. Microscope, infrared spectrum analysis and X-ray diffraction analysis showed that the inclusion compound was formed and the drug existed in the inclusion compound as amorphous. Conclusion: The inclusion compound of silymarin-SBE-β-CD has been successfully prepared, which can significantly improve the solubility of the drug, and provide experimental basis for its clinical application.
10.Transcriptome analysis reveals the role of withering treatment in flavor formation of oolong tea (Camellia sinensis).
Chen ZHU ; Shuting ZHANG ; Chengzhe ZHOU ; Biying SHI ; Linjie HUANG ; Yuling LIN ; Zhongxiong LAI ; Yuqiong GUO
Chinese Journal of Biotechnology 2022;38(1):303-327
Oolong tea is a semi-fermented tea with strong flavor, which is widely favored by consumers because of its floral and fruity aroma as well as fresh and mellow taste. During the processing of oolong tea, withering is the first indispensable process for improving flavor formation. However, the molecular mechanism that affects the flavor formation of oolong tea during withering remains unclear. Transcriptome sequencing was used to analyze the difference among the fresh leaves, indoor-withered leaves and solar-withered leaves of oolong tea. A total of 10 793 differentially expressed genes were identified from the three samples. KEGG enrichment analysis showed that the differentially expressed genes were mainly involved in flavonoid synthesis, terpenoid synthesis, plant hormone signal transduction and spliceosome pathways. Subsequently, twelve differentially expressed genes and four differential splicing genes were identified from the four enrichment pathways for fluorescence quantitative PCR analysis. The results showed that the expression patterns of the selected genes during withering were consistent with the results in the transcriptome datasets. Further analysis revealed that the transcriptional inhibition of flavonoid biosynthesis-related genes, the transcriptional enhancement of terpenoid biosynthesis-related genes, as well as the jasmonic acid signal transduction and the alternative splicing mechanism jointly contributed to the flavor formation of high floral and fruity aroma and low bitterness in solar-withered leaves. The results may facilitate better understanding the molecular mechanisms of solar-withering treatment in flavor formation of oolong tea.
Camellia sinensis/genetics*
;
Gene Expression Profiling
;
Plant Leaves
;
Plant Proteins/metabolism*
;
Taste
;
Tea
;
Transcriptome/genetics*