Isochrysis galbana is considered an ideal bait for functional foods and nutraceuticals of humans because of its high fucoxanthin(Fx)content.However,multi-omics analysis of the regula-tory networks for Fx biosynthesis in I.galbana has not been reported.In this study,we report a high-quality genome assembly of I.galbana LG007,which has a genome size of 92.73 Mb,with a contig N50 of 6.99 Mb and 14,900 protein-coding genes.Phylogenetic analysis confirmed the monophyly of Haptophyta,with I.galbana sister to Emiliania huxleyi and Chrysochromulina tobinii.Evolutionary analysis revealed an estimated divergence time between I.galbana and E.huxleyi of～133 million years ago.Gene family analysis indicated that lipid metabolism-related genes exhibited significant expansion,including IgPLMT,IgOAR1,and IgDEGS1.Metabolome analysis showed that the content of carotenoids in I.galbana cultured under green light for 7 days was higher than that under white light,and β-carotene was the main carotenoid,accounting for 79.09％of the total carotenoids.Comprehensive multi-omics analysis revealed that the content of β-carotene,antheraxanthin,zeaxanthin,and Fx was increased by green light induction,which was significantly correlated with the expression of IgMYB98,IgZDS,IgPDS,IgLHCX2,IgZEP,IgLCYb,and IgNSY.These findings contribute to the understanding of Fx biosynthesis and its regulation,pro-viding a valuable reference for food and pharmaceutical applications.

【Objective】 To develop a new approach for the preparation of 0.7~0.8 hematocrit concentrated washed red blood cells(RBCs) for fetal anemia in utero transfusion and apply it in clinical. 【Methods】 The erythrocyte suspension and frozen stored erythrocytes within expiry date in Guangzhou Blood Center from March 2020 to February 2021 were taken to prepare concentrated washed RBCs. According to the derivation formula, corresponding weight of RBC preservation solution was added to obtain 0.7~0.8 hematocrit concentrated washed RBCs. Routine blood test data were statistically analyzed by single-sample t test, and P<0.05 was considered statistically significant. Qualified Rh-negative/ O-type 0.7~0.8 hematocrit concentrated RBCs within expiry date were used in clinical intrauterine blood transfusion. 【Results】 The hematocrit of concentrated washing RBCs prepared by the new approach could reach 0.7~0.8. The RBCs count (8.389 ±0.808)×1 0¹²/ L and hemoglobin content(233.730±15.498)g/L were higher while the erythrocyte count (0.732±0.469)×10⁹ /L and platelets count(26.000±26.276)×10⁹/L were lower than the normal values of adults. The mean erythrocyte volume(fL), hemoglobin content(pg) and concentration(g/L )were 88.123±6.359, 30.004±2.809 and 339.980±11.865, respectively, which were normal values of adults. Fetal anemia was significantly improved and the prognosis was good after intrauterine blood transfusion. 【Conclusion】 The 0.7~0.8 hematocrit concentrated washed RBCs prepared by the new approach is consistent with the special blood requirements during fetal anaemia transfusion, meets the clinical treatment standards, and can be applied in clinical.

In order to ensure the normal teaching order during the prevention and control of the COVID-19 epidemic, the second semester of the 2019-2020 academic year in Central South University was devoted to the online teaching. In response to the school's call, the diagnostics teaching team has applied the Tencent classroom software, WeChat mini programs, analog teaching software and digital curriculum platform to carry out online teaching activities. On the basis of summarizing the previous online teaching experience, we have made a preliminary discussion and reflection on the online teaching, which will provide ideas and directions for the reform of medical education.

Objective:To evaluate the clinical efficacy of multi-disciplinary single center's CCCG-HB-2016 regimen in the treatment of hepatoblastoma (HB) in children.Methods:Clinical data of 36 HB patients treated with CCCG-HB-2016 program from Aug 2016 to March 2020 were analyzed.Results:These 36 patients included 20 boys and 16 girls. The serum AFP was all higher than 2 792 ng/ml,there was a correlation between AFP and tumor risk stratification ( H=14.973, P<0.05). Twenty eight cases (77.78%) were epithelial type and 8 cases (22.22%) were mixed epithelial mesenchymal type.All children were treated by tumor resection combined with chemotherapy, and there was a correlation between tumor risk stratification and surgical resection of liver lobe ( H=8.847, P<0.05). The probability of bone marrow suppression in the low-risk group was 58.33% (35/60),that in the intermediate-risk group was 73.49% (61/83) and in the high-risk group was 80.23% (69/86).All 36 cases were followed up to March 31, 2020,with an average follow-up of 21.9 months and the median survival was 22.5 months.The overall survival rate (OS) and event-free survival rate (EFS) were 97.2% and 83.3% respectively. Conclusions:The multidisciplinary CCCG-HB-2016 regimen was with a high success rate and along with a high incidence of bone marrow suppression.

Objective:Based on ear cartilage of congenital microtia, we tried to find the different metabolites and variational metabolic pathways on molecular level by gas chromatography time-of-flight mass spectrometry (GC-TOFMS).Methods:Patients with microtia were collected from June 2017 to January 2018. During operation, the ear cartilage tissue was collected and labeled, and the case group was residual ear cartilage tissue of the patients with microtia, with 18 cases. The control group was normal ear cartilage tissue, total 18 cases. After the sample pretreatment, based on the technique of GC-TOFMS and the software of XploreMET, the differential metabolites in the residual ear cartilage of microtia were analyzed by orthogonal partial least square discriminant analysis (OPLS-DA) and its variable importance projection (VIP) and U test statistical method. Compared with the metabolite database, the metabolic pathway enrichment analysis (MPEA) was used to screen the most related metabolic pathways. Results:According to the multidimensional statistical analysis, it is different in the metabolites of ear cartilage between the two groups. According to the verification of single dimensional statistical analysis, 14 specific biomarkers were identified, and their P value was less than 0.05. According to the metabolic pathway enrichment analysis (MPEA), 3 metabolic pathways had statistically significant difference, including arginine metabolism, taurine and the taurine metabolism, pantothenate and CoA metabolism, beta-alanine metabolism, glycerophospholipid metabolism, P <0.05. Conclusions:Arginine, taurine, L-cysteine and pantothenic acid may have an association with microtia, which proved that it is feasible to study the pathogenesis of microtia by metabonomics.

Objective:To investigate the microenvironment associated with tumorimmunity in regenerated lymph nodes treated with VEGF-C, which involves the lymphangiogenesis, chemokine as well as alteration of cell populations.Methods:A total of 10 nude mice were randomized to 2 groups. 5 mice in group A were treated with autologous lymph nodes fragmentary transplantation together with VEGF-C, 5 mice without any intervention in group B were served as control group. 4 weeks after surgeries, MDA-MB-231-Luc-GFP cells (5×10 6 cells/mouse, 100 μl of 40% matrigel solution) were injected subcutaneously in the thoracic flunk of nude mice in group A, the same number cells were implanted into the second mammary fat pad in group B as well. 4 weeks post inoculation, the tumor draining lymph nodes (TDLNs) in both groups were harvested, and the microenvironment was assessed by hematoxylin-eosin (HE) staining and multispectral immunofluorescence staining for LYVE-1, chemokine CCL21, programmed death ligand-1(PD-L1) and F4/80. Results:The specific markers were analyzed based on positive cell percentage between group A and group B. The LYVE-1 and F4/80 expression in the group A was significantly higher than that in group B[LYVE-1: group A 21.44 0(34.675)% vs group B 2.964 (4.160)%, P<0.001; F4/80: group A 5.396 (7.205)% vs group B 3.573 (2.670)%, P=0.020)], and there were no significant differences in the expression of CCL21 and PD-L1 between these two groups [CCL21: group A 21.470((29.145)% vs group B 16.430((29.145)%( P=0.166); PD-L1: group A10.000 (14.430)%vs group B 4.070 (26.740)%, P=0.091]. Additionally, the double positive expression of LYVE-1/CCL21 and LYVE-1/F4/80 had still statistically significant differences[LYVE-1/CCL21: group A 8.637 (13.150)% vs group B 1.571 (1.680)%, P<0.001; LYVE-1/F4/80: group A 6.181 (9.050)% vs group B 1.454 (0.830)%, P<0.001], whereas, the two groups did not differ from each other in the double positive expression of LYVE-1/PD-L1 [group A 3.617 (4.195)% vs group B 2.363 (3.900)%, P=0.266]. Conclusions:The renewed lymph nodes treated with VEGF-C demonstrate some more immune suppressive conditions such as lymphatic endothelial cells hyperplasia, myeloid-derived suppressor cells infiltration, enhanced expression of CCL21 and PD-L1.

Objective:Based on ear cartilage of congenital microtia, we tried to find the different metabolites and variational metabolic pathways on molecular level by gas chromatography time-of-flight mass spectrometry (GC-TOFMS).Methods:Patients with microtia were collected from June 2017 to January 2018. During operation, the ear cartilage tissue was collected and labeled, and the case group was residual ear cartilage tissue of the patients with microtia, with 18 cases. The control group was normal ear cartilage tissue, total 18 cases. After the sample pretreatment, based on the technique of GC-TOFMS and the software of XploreMET, the differential metabolites in the residual ear cartilage of microtia were analyzed by orthogonal partial least square discriminant analysis (OPLS-DA) and its variable importance projection (VIP) and U test statistical method. Compared with the metabolite database, the metabolic pathway enrichment analysis (MPEA) was used to screen the most related metabolic pathways. Results:According to the multidimensional statistical analysis, it is different in the metabolites of ear cartilage between the two groups. According to the verification of single dimensional statistical analysis, 14 specific biomarkers were identified, and their P value was less than 0.05. According to the metabolic pathway enrichment analysis (MPEA), 3 metabolic pathways had statistically significant difference, including arginine metabolism, taurine and the taurine metabolism, pantothenate and CoA metabolism, beta-alanine metabolism, glycerophospholipid metabolism, P <0.05. Conclusions:Arginine, taurine, L-cysteine and pantothenic acid may have an association with microtia, which proved that it is feasible to study the pathogenesis of microtia by metabonomics.

Objective:To investigate the microenvironment associated with tumorimmunity in regenerated lymph nodes treated with VEGF-C, which involves the lymphangiogenesis, chemokine as well as alteration of cell populations.Methods:A total of 10 nude mice were randomized to 2 groups. 5 mice in group A were treated with autologous lymph nodes fragmentary transplantation together with VEGF-C, 5 mice without any intervention in group B were served as control group. 4 weeks after surgeries, MDA-MB-231-Luc-GFP cells (5×10 6 cells/mouse, 100 μl of 40% matrigel solution) were injected subcutaneously in the thoracic flunk of nude mice in group A, the same number cells were implanted into the second mammary fat pad in group B as well. 4 weeks post inoculation, the tumor draining lymph nodes (TDLNs) in both groups were harvested, and the microenvironment was assessed by hematoxylin-eosin (HE) staining and multispectral immunofluorescence staining for LYVE-1, chemokine CCL21, programmed death ligand-1(PD-L1) and F4/80. Results:The specific markers were analyzed based on positive cell percentage between group A and group B. The LYVE-1 and F4/80 expression in the group A was significantly higher than that in group B[LYVE-1: group A 21.44 0(34.675)% vs group B 2.964 (4.160)%, P<0.001; F4/80: group A 5.396 (7.205)% vs group B 3.573 (2.670)%, P=0.020)], and there were no significant differences in the expression of CCL21 and PD-L1 between these two groups [CCL21: group A 21.470((29.145)% vs group B 16.430((29.145)%( P=0.166); PD-L1: group A10.000 (14.430)%vs group B 4.070 (26.740)%, P=0.091]. Additionally, the double positive expression of LYVE-1/CCL21 and LYVE-1/F4/80 had still statistically significant differences[LYVE-1/CCL21: group A 8.637 (13.150)% vs group B 1.571 (1.680)%, P<0.001; LYVE-1/F4/80: group A 6.181 (9.050)% vs group B 1.454 (0.830)%, P<0.001], whereas, the two groups did not differ from each other in the double positive expression of LYVE-1/PD-L1 [group A 3.617 (4.195)% vs group B 2.363 (3.900)%, P=0.266]. Conclusions:The renewed lymph nodes treated with VEGF-C demonstrate some more immune suppressive conditions such as lymphatic endothelial cells hyperplasia, myeloid-derived suppressor cells infiltration, enhanced expression of CCL21 and PD-L1.

Objective: Autologous lymph nodes fragmentary transplantation combined with vascular endothelial growth factor-C (VEGF-C) on athymic nude mice to explore the association between regeneration of lymphatic vessel and tumor cell migration. Methods: A total of 45 nude mice were randomly divided into 3 groups: Group A, simple autologous lymph nodes fragmentary transplantation, n=15; Group B, autologous lymph nodes fragmentary transplantation together with VEGF-C, n=15; Group C, without any intervention, n=15. At 1 month, 2 months and 6months after surgeries, the axillary lymph nodes of 5 mice in each group were dynamically examined by in vivo indocyanine green (ICG) fluorescence imaging respectively. The regenerated lymph nodes and relevant skin were evaluated using hematoxylin-eosin (H&E) staining and the skin was quantitatively analyzed via immunofluorescence staining for lymphatic vessel endothelial hyaluronan receptor 1(LYVE-1) as well. Results: One month after surgery, the right regenerated axillary lymph nodes in group B (5/5) were visible by in vivo ICG fluorescence imaging, whereas the same signals were not detected in group A (0/5). The results were the same at 2 and 6 months after surgery. HE staining showed that the cortical, paracortical, and medullary regions of the right axillary lymph nodes of the experimental group B were clear, and the lymphatic vessel structure was present, accompanied by lymphocyte infiltration. Immunofluorescence staining of the right upper limb showed that the expression of LYVE-1 in the lymphatic endothelium of the B group was significantly higher than that in group A (P<0.001) and the control group (both P<0.001). Conclusions Due to the promising consequence of regenerated lymph nodes, the procedure of autologous lymph nodes fragmentary transplantation combined with VEGF-C in athymic nude mice provides a reliable animal model for the next stage research.

Objective The research about the effect of different chemotherapeutic drugs on CD19?CAR?T cells with CCK8 test to provide the theoretical support about the development of chemotherapy for clinical support. Methods Extract T cells from a normal adult peripheral blood and synthesize CD19?CAR?T cell. CD19?CAR?T cells were treated with different doses of chemotherapeutic drugs for 24,48,72 h and(or)96 h,and inhibition rate was calculated. Results First,we observed that the inhibition rates of fludarabine and Mafosfamide for CD19?CAR?T cells were increasing with the time and concentration (P < 0.05). Secondly , Bus ulfan had no effect in CD19?CAR?T cells in vitro(P > 0.05). Finally ,Cyclophosphamide had no effect in CD19?CAR?T cells in vitro (P > 0.05). Conclusion Mafosfamide and Fludarabine can inhibit the CD19?CAR?T cells. Cyclophosphamide have no activity in vitro.