The aim of this study was to promote fucoxanthin accumulation in Phaeodactylum tricornutum by photo-fermentation through optimizing the mode of multiple nitrogen supplementation and blue light enhancement. The results showed that the mixed nitrogen source (tryptone: urea=1:1, N mol/N mol; total nitrogen concentration at 0.02 mol/L) added to the culture system by six times was the best mode in shake flasks. Two-phase culture with light adjustment was then carried out in 5 L photo-fermenter with an enhanced blue light (R: G: B=67.1:16.7:16.3) in the second phase, leading to improved cell density (1.12×10⁸ cells/mL), biomass productivity (330 mg/(d·L)), fucoxanthin content (19.62 mg/g), titer (69.71 mg/L) and productivity (6.97 mg/(d·L)). Compared with one-phase culture under red/blue (R: G: B=70.9:18.3:10.9) light and six-times nitrogen supplementation, the fucoxanthin content was significantly increased by 7.68% (P < 0.05) but the productivity did not change significantly (P > 0.05). Compared with one-phase culture under red/blue (R: G: B=70.9:18.3:10.9) light and one-time nitrogen supplementation, the content and productivity of fucoxanthin were significantly increased by 45.98% and 48.30% (P < 0.05), respectively. This study developed a two-phase culture mode with multiple nitrogen supplementation and blue light enhancement, which effectively promoted the accumulation of fucoxanthin and improved the efficiency of nitrogen source utilization, thus providing a new approach for fucoxanthin accumulation in P. tricornutum by photo-fermentation.
Nitrogen ; Light ; Xanthophylls ; Diatoms ; Dietary Supplements

The aim of this study was to develop a technical system for high-efficient production of fucoxanthin by photo-fermentation of Phaeodactylum tricornutum. In a 5 L photo-fermentation tank, the effects of initial light intensity, nitrogen source and concentration as well as light quality on biomass concentration and fucoxanthin accumulation in P. tricornutum were investigated systematically under mixotrophic condition. The results showed that the biomass concentration, fucoxanthin content and productivity reached the highest level of 3.80 g/L, 13.44 mg/g and 4.70 mg/(L·d) under the optimal conditions of initial light intensity of 100 μmol/(m²·s), 0.02 mol TN/L of tryptone: urea (1:1, N mol/N mol) as mixed nitrogen source, and a mixed red/blue (R: B=6:1) light, 1.41, 1.33 and 2.05-fold higher than that before optimization, respectively. This study developed a key technology for enhancing the production of fucoxanthin by photo-fermentation of P. tricornutum, facilitating the development of marine natural products.
Fermentation ; Xanthophylls ; Light ; Diatoms ; Nitrogen

嵌合抗原受体T（chimeric antigen receptor T， CAR-T）细胞是一种通过基因工程表达受体的T细胞，能够识别特定的抗 原，是目前最具潜力的靶向肿瘤治疗方法。然而，作为抗癌免疫系统中主要效应细胞之一的CD8+T细胞在肿瘤中发挥作用时， 通 常处于耗竭状态，而这种功能缺陷的CD8+T细胞是杀伤肿瘤的障碍。肿瘤微环境（tumor microenvironment，TME）中存在许多抑 制性因素，例如耗竭性T细胞表面高表达的抑制性受体、免疫抑制细胞群、抑制性因子、转录因素、代谢因素等都对T细胞的分化 及耗竭有重要影响。当然， CAR的结构和共刺激域也对CAR-T细胞整体功能发挥着重要作用。本文着重总结近年有关CD8+T 细胞耗竭的机制及改善策略的研究进展，为增强CAR-T细胞的抗肿瘤效应提供了潜在思路。

Objective To improve the quality of clinical biochemistry laboratory by quality indicators of pre-analytical,analytical,post-analytical phase and the whole process.Methods Analytical Phase:The Sigma values of items were calculated,applying the equation Sigma =(TEa％-Bias％)/CV％.Total allowable error (TEa) is from analyticalal specification defined in WS/T403-2012 of China,Bias％ is from the evaluation results of National Center for Clinical Laboratory (NCCL) trueness verification PT series and CV％ is from internal quality control data during the last 6 months in our lab.Normalized Sigma metrics plot was made to evaluate the analysis performance and the quality control strategies were designed accordingly.The quality goal indexes (QGI) were also calculated to propose improvement measures for items below 6 Sigma.Quality indicators of pre-,post-analytical and whole analytical phase,such as quality of specimen,critical value notification,critical value notification in time,TAT of hs-cTnT,TAT of emergency biochemical items,rewrite of laboratory reports and unacceptable performance in EQA-PT were measured in Sigma metrics too.The Sigma metrics changes before and after taking improvement measures were compared to conform the effectiveness.Results The average Sigma value of 17 biochemical tests was 5.29,of which 8 items (UA,K,ALP,CK,AMY,AST,TG,Na) achieved excellent to world class level (≥ 5 Sigma),6 items (LDH,Cre,TC,ALT,Mg,Glu) achieved marginal to good level (5 ＞ Sigma ≥ 3),BUN performed poorly (3 ＞ Sigma ≥ 2),Ca,TP performed unacceptably (Sigma ＜ 2) with serious quality defects.The Sigma values of unacceptable specimen,critical value notification,critical value notification in time,unacceptable turn around time (TAT) of hs-cTnT,unacceptable turn around time (TAT) of emergency biochemical items,rewrite of laboratory reports,unacceptable performance in EQA-PT were 4.17,3.60,2.75,1.72,3.27,4.52,3.33 respectively,rising to 4.30,4.30,2.90,2.45,3.75,4.80,3.60 accordingly after improvement.Conclusions Sigma metrics is potentially an ideal approach for clinical biochemistry laboratories management,which is helpful to find out problems,put forward improvement measures,and confirm the effectiveness,so as to achieve the purpose of continuous quality improvement.

Objective To test the hypothesis that the CCL2/CCR2 signaling pathway plays an important role in pain induced by experimental tooth movement.Methods Male Sprague-Dawley rats weighing between 200 and 300 g were used in this study.Expression of CCL2/CCR2 in the trigeminal ganglion(TG) was determined by Western blotting 0 h,4 h,1 d,3 d,5 d,7 d after tooth movement.Localization of the CCL2 was revealed by immunohistochemistry.Changes in body weight,nocifensive behaviors,and the effects of CCL2/CCR2 antagonists on these changes in pain behaviors were evaluated.Exogenous CCL2 was injected into periodontal tissues and added to TG neurons in culture and the resulting c-fos expression and pain responses were detected.In addition,the expression and cellular localization of CCL2 in the medullary dorsal horn (MDH) was determined by immunohistochemistry 3 d and 14 d after tooth movement.Results Experimental tooth movement led to a statistically significant increase in CCL2/CCR2 expression at the protein level from day 3 to 7 after application of force initiating tooth movement.When compared with control group(1.000± 0.000),CCL2 increased to (2.620 ± 0.128),(3.300±0.197) and (1.740±1.290) at day 3,5 and 7 respectively,which were statistically significant (P＜0.05).CCR2 expression levels were (1.636±0.061) and (1.766±0.126) compared with that in control group (1.000±0.000) at day 3 and 5 respectively with statistical significance (P＜0.05).Both of them peaked on day 5 (3.3 and 1.8 time compared to control group).Application of recombinant CCL2 led to the up-regulation of c-fos expression in vivo and in vitro,and triggered a corresponding nocifensive behavior in rats.The magnitude of the nocifensive behavior could be reduced by a CCR2 antagonist,and by CCL2 neutralizing antibody.Furthermore,we found a significant increase in the expression of CCL2,corresponding well to the upregulation of the time spent on nocifensive behaviors after ETM.In addition,CCL2 was up-regulated in TG neurons and astrocytes in Vc.Conclusions The CCL2/CCR2 axis was modulated by experimental tooth movement and involved in the development of tooth movement pain,and thus palyed an important role in orthodontic pain mechanism.

Objective To assess the two-dimensional carotid strain as an index of arterial stiffness in patients without carotid atherosclerotic plaques,and its clinic value in cardiovascular risk stratification.Methods All patients were divided into three groups (low risk,intermediate risk,high risk group) by Framingham cardiovascular risk scores,two-dimensional carotid circumferential strain (CS),carotid intimamedia thickness (IMT) were evaluated.CS was adjusted for pulse pressure (CS/PP).Results CS,CS/PP,IMT were significant difference between low risk and intermediate risk groups,low risk and high risk groups (P ＜0.05).IMT and CS were not significant difference between intermediate risk and high risk group (P =0.23,P =0.57).CS/PP was significant difference between three groups (P ＜0.05).Both CS and CS/PP were correlated with IMT (r =-0.30,r =-0.33,P ＜0.05).Conclusions Two dimensional strain could assess the carotid arterial mechanics.IMT and CS could evaluate the structural and functional alternations of carotid stiffness.Combining these two indices allowed more accurate evaluation of the subclinical phase of the atherosclerotic disease.

OBJECTIVETo investigate the frequency of anti-Mur and Mur antigen among blood donors in Guangzhou to provide evidence for guiding clinical transfusion and prenatal screening.

METHODSDG Gel Coombs cards were used to screen active anti-Mur at 37 degrees celsius; from 2725 blood donors. Multiplex ligation-dependent probe amplification (MLPA) was used to genotype Mur antigen from 91 blood donors, and human anti-Mur serum was used to verify the phenotypes deduced from the genotypes.

RESULTSThe frequency of anti-Mur and genotyped Mur antigen was 0.04% (1/2725) and 6.59% (6/91), respectively, and the phenotyping results were consistent with the genotyping results.

CONCLUSIONThe blood donors in Guangzhou show a low frequency of anti-Mur and a relatively high frequency of Mur antigen. Genotyping using MLPA allows Mur antigen genotyping when commercial anti-Mur is not available.

Blood Donors ; Blood Group Antigens ; genetics ; immunology ; China ; Genotype ; Genotyping Techniques ; Humans ; Multiplex Polymerase Chain Reaction ; Phenotype

OBJECTIVETo report a rare case of hemolytic disease of the newborn (HDN) with kernicterus caused by anti-Di(a) diagnosed using high-throughput genotyping multiplex ligation-dependent probe amplification (MLPA).

METHODSConventional serological methods were used to detect the antibodies related with HDN. The genotypes of more than 40 red blood cell antigens for the newborn and her parents were obtained using the high-throughput MLPA assay. The antibody titers were tested using a standard serological method.

RESULTSThe unknown antibody against the low-frequency antigens was predicted based on the primary serological tests. The genotyping results for more than 40 red blood cell antigens of the newborn and her parents showed incompatible antigens of MNS and Diego blood group system, indicating the existence of anti-N or anti-Di(a). Further serological tests confirmed anti-Di(a) existence in the plasma of the newborn and her mother. The titer of anti-Di(a) in the mother's plasma was 1:32.

CONCLUSIONSevere HDN including kernicterus can result from anti-Di(a). High-throughput genotyping MLPA assay can help type some rare antigens in complicated cases. The reagent red cell panels including Di(a)-positive cells are necessary in routine antibody screening test in Chinese population.

Blood Group Incompatibility ; genetics ; Erythroblastosis, Fetal ; diagnosis ; immunology ; Exchange Transfusion, Whole Blood ; Female ; Genotype ; Humans ; Infant, Newborn ; Nucleic Acid Amplification Techniques ; methods ; Rh-Hr Blood-Group System ; genetics ; immunology ; Rho(D) Immune Globulin ; genetics ; immunology

ObjectiveTo investigate the ability of three-dimensional(3D) speckle tracking imaging (STI) for assessing left ventricular (LV) strain,and measure the value and angle of myocardial main strain vector.MethoodsLongitudinal,circumferential and radial strain of LV were measured in 31 healthy adults by 3D-STI and two-dimensional(2D) STI.The main strain vector and the angle between main strain vector and the LV short-axis plane were calculated from longitudinal and circumferential strains.Results The global longitudinal,circumferential strains and strains at basal,middle,apical level of LV by 3D-STI were significantly smaller than those by 2D-STI,whereas radial strain showed an opposite trend.The 3D strains presented significant difference between different levels of LV,longitudinal,circumferential and radial 3D strains were greater at middle level than at basal and apical level of LV,meantime 2D stain didn't show such obvious trend.Calculated from three-dimensional longitudinal and circumferential strain,the main strain vector was greatest at middle level of LV,whereas the angle between main strain vector and LV short axis plane didn't show any significant difference between three levels of LV.Conclusions3D-STI provides us a new and reproducible method for the evaluation of LV regional function by measuring LV strains.

ObjectiveTo study the molecular genetic mechanism of para-bombay phenotype in two individuals.MethodsThe proband was a female.When the proband donated blood,because the forward blood group wasn't coincident with her reverse blood group,the blood and saliva specimen from proband and her family members were sent to Guangzhou Blood Center for further identification.Routine serological techniques were used to determine proband's and her family members' blood group and ABH antigen in saliva.The coding regions of FUT1 and FUT2 gene,exon 6 and exon 7 of ABO gene were amplified by polymerase chain reaction using proband's and her family members' genomic DNA.All amplified products were analyzed after being directly sequenced.The two-base deletion regions of FUT1 gene were certified by cloning and haplotype sequencing.Results Proband's and her little brother's blood group were identified as para-bombay while other family members' blood group were normal.Two-base deletion heterozygous mutations of FUT1 gene were found in proband and her brother,AG deletion at position 547-552 and TT deletion at position 880-882,which caused a reading frame shift and a premature stop eodon.Meanwhile,880-882del TT heterozygous mutation was found in proband's grandfather and her father and 547-552del AG heterozygous mutation was found in proband's mother and her little sister.ResultsOf cloning and haplotype sequencing certified that these two-base deletion mutations occurred at 547-548 and 881-882 position respectively.Three new mutations were found in FUT2 gene,390C ＞ T,418A ＞ T and 749G ＞ A,which could cause the change of amino acid at position 140Ile ＞ Phe and 250Arg ＞ Gln.Conclusions Two-base deletion heterozygous mutations in different positions in FUT1 gene were found in 2 individuals,which maybe the molecular genetic mechanism of para-bombay phenotype.Heterozygous deletion mutation in one-strand DNA wouldn't change the ABO blood group.Three new mutations were also found in FUT2 gene.( Chin J Lab Med,2012,35:815-819)