Cervical cancer with transferrin receptor has a poor prognosis and is associated with immune infiltration, according to a comprehensive bioinformatics study

Background:The molecular mechanism underlying the involvement of the Transferrin receptor(TFRC)in cervical cancer remains poorly understood.This study aims to elucidate the role of TFRC in cervical cancer by analyzing data from The Cancer Genome Atlas(TCGA)and Genotype-Tissue Expression(GTEx)databases.Methods:TFRC protein expression was obtained from Human Protein Altas(HPA).All datas were collected from TCGA and GTEx.In this study,we analyzed the expression of TFRC in cervical cancer and its clinical significance.Through Kyoto Encyclopedia of Genes and Genomes(KEGG)and Gene set enrichment analyses(GSEA),investigated the related molecular pathways of TFRC.The relationship between TFRC and immune infiltration was then examined.The prognosis of different immune cell subsets was then analyzed after dividing cervical cancer patients into high and low expression of TFRC groups.Results:TFRC is highly expressed in various tumor tissues compared to control normal tissues,including cervical cancer.An increased expression of TFRC was associated with higher Tumor(T)and Node(N)stage,as well as a higher clinical stage.Kaplan–Meier(KM)survival analysis investigated that higher TFRC expression patients have a poor overall survival(OS),disease specific survival(DSS)and progress free interval(PFI).Both KEGG and GSEA enriched signaling pathway by high TFRC and low TFRC groups.There was a significant negative linear correlation between TFRC expression and immune infiltration.TFRC affects the prognosis of cervical cancer patients through immune pathway.Conclusions:Cervical cancer patients with TFRC expression may have a worse prognosis.

Effect of Insulin-Transferrin-Selenium on Goat Oocytes Maturation and Embryo Development

[Objective] The research aimed to enhance culture efficiencies of oocyte and embryo of goat in vitro and to explore serum-free culture system in vitro.[Method] At present,the conventional solutions of oocyte maturation and embryo development in vitro were always added into 1% ITS(Insulin-transferrin-selenium) or using 1% ITS to replace FBS in 2 kinds culture solutions for conducting in vitro cultures of goat oocyte and parthenogenetic embryo.The influences of ITS on their developments were detected.[Result] ITS in maturation liquid of oocytes could not increase oocytes maturation rate but significantly increased blastocyst rate (58.06% vs. 48.19%)of parthenogenetic embryo.If FBS in maturation liquid of oocytes was replaced by ITS, the maturation rate, cleavage rate and blastocyst rate were basically unchanged.Adding ITS into embryo medium could increase blastocyst rate (68.30% vs. 56.82%)of parthenogenetic embryo of goat.If FBS in embryo medium was replaced by ITS,the cleavage rate didn’t change basically,while the blastocyst rate in ITS was obviously lower than that in FBS group(42.33% vs.56.82%).[Conclusion] ITS could promote maturation of oocyte in vitro and early embryonic development, in addition,ITS could replace serum in maturation medium of oocyte as serum-free culture system for conducting relevant researches.

Expression of transferrin in hematoma brain tissue at different stages after intra cerebral hemorrhage in rats

Objective: To explore the expression of transferrin(Tf) and transferrin receptor(Tf R) in hematoma brain tissue at different stage after intracerebral hemorrhage(ICH) in rats. Methods: ICH rats model were established by collagenase method, and rats were sacrificed at 24 h, 72 h, 7 d and 14 d after operation. The levels of Tf and Tf R in different periods of rats were detected by immunohistochemical method, and correlation between two groups was analyzed. Results: Tf, Tf R-positive cells at each time after operation in observation group were significantly higher than that in control group(P<0.05). Tf, Tf R-positive cells began to increase from 24 h after the operation and reached the peak 72 h-7 d after surgery, but then gradually decreased. Tf was mainly expressed in nucleus and cytoplasm of neurons and glial cells around the hematoma, but Tf R was mainly expressed in nucleus and cytoplasm of neurons and choroid plexus endothelial cells. Correlation analysis showed that the Tf-positive cell was significantly positively correlated with Tf R-positive cell expression(r=0.447, P=0.022). Conclusions: Tf and Tf R were important transporters in brain tissue excessive load iron transport after ICH, and detecting the expression levels of the two indicators can provide a reference for prognosis treatmentin ICH.

Transferrin receptor and ferritin-H are developmentally regulated in oligodendrocyte lineage cells

Iron is an essential trophic element that is required for cell viability and differentiation, especially in oligodendrocytes, which consume relatively high rates of energy to produce myelin. Multiple iron metabolism proteins are expressed in the brain including transferrin receptor and ferritin-H. However, it is still unknown whether they are developmentally regulated in oligodendrocyte lineage cells for myelination. Here, using an in vitro cultured differentiation model of oligodendrocytes, we found that both transferrin receptor and ferritin-H are significantly upregulated during oligodendrocyte maturation, implying the essential role of iron in the development of oligodendrocytes. Additional different doses of Fe3＋ in the cultured medium did not affect oligodendrocyte precursor cell maturation or ferritin-H expression but decreased the expression of the transferrin receptor. These results indicate that upregulation of both transferrin receptor and ferritin-H contributes to maturation and myelination of oligodendrocyte precursor cells.

Construction of single chain Fv antibody against transferrin receptor and its protein fusion with alkaline phosphatase

AIM: To construct fusion protein of a single-chain antibody (scFv) against transferrin receptor (TfR) with alkaline phosphatase(AP). METHODS: The VH-linker-VL,namely scFv gene,was prepared by amplifying the VH and VL genes from plasmid pGEM-T-VH and pGEM-T-VL with splicing overlap extension polymerase chain reaction (SOE PCR). After the ScFv gene was modified by 5/71 and Not I,it was subcloned into the secretory expression vector pUC19/119, and then was transformed into E.coli TG1.The positive colonies were screened by colony PCR and their expressions were induced by IPTG.ScFv gene was gained by digesting ScFv expression vector pUC19/119 with 5/71 and NotI restriction enzymes, then subcloned into expression vector pDAP2, followed by transformation in E.coli TG1.The positive colonies were selected by bacterial colony PCR.The expression of fusion protein (scFv-AP) was induced by IPTG.Its activity was detected by enzyme immunoassay. The molecular weights of scFv and scFv-AP were measured by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). RESULTS: The product of SOE PCR formed a band of 700 bp in agarose gel electrophoresis. SDS-PAGE demonstrated the molecular weight of scFv was 27 ku.Immunofluorescent assay (IFA) demonstrated its reactivity with TfR.The molecular weight of scFv-AP was 75 ku.Enzyme immunoassay showed that scFv-AP could specifically bind to human TfR and play AP activity. CONCLUSION: We have successfully prepared the anti-human TfR scFv and constructed the fusion protein of scFv and AP.It is promising for immunological experiments.

Study on The Method of Quantitative Analysis of Serum Ferritin and Soluble Transferrin Receptor with Protein Microarray Technology

Objective To establish and evaluate a protein serum ferritin （SF） and soluble transferrin receptor microarray method for combined measurement of （sTfR）. Methods Microarrayer was used to print both anti-SF antibodies I and anti-sTfR antibodies I on each protein microarray. Anti-SF antibodies II and anti-sTfR antibodies II were used as detection antibodies and goat antibodies coupled to Cy3 were used as antibodies Ill. The detection conditions of the quantitative analysis method for simultaneous measurement of SF and sTfR with protein microarray were optimized and evaluated. The protein microarray was compared with commercially available traditional tests with 26 serum samples. Results By comparison experiment, mouse monoclonal antibodies were chosen as the probes and contact printing was chosen as the printing method. The concentrations of SF and sTfR probes were 0.5 mg/mL and 0.5 mg/mL respectively, while those of SF and sTfR detection antibodies were 5 μg/mL and 0.36 μg/mL respectively. Intra- and inter-assay variability was between 3.26% and 18.38% for all tests. The regression coefficients comparing protein microarray with traditional test assays were better than 0.81 for SF and sTfR. Conclusion The present study has established a protein microarray method for combined measurement of SF and sTfR.

Binding Constants for Terbium(Ⅲ) with Chicken Apoovotransferrin

The binding of Tb 3+ to chicken apoovotransferrin was studied by monitoring the fluorescent intensity of Tb 3+ at 549 nm. The conditional equilibrium constants for the complexation of Tb 3+ by chicken apoovotransferrin in 0 1 mol/L hepes, at pH 7 4 and room temperature were measured. The successive macroscopic binding constants are lg K 1=9 08±0 12 and lg K 2=7 36±0 22. The molar fluorescence enhancement of Tb 3+ apoovotransferrin complex is (2 06±0 14)×10 4 mol -1 ·L. The fluorescence quenching experiment and the titration of N terminal monoferric ovotransferrin showed that Tb 3+ has a preference for being bound to the N terminal binding site of apoovotransferrin.

miR-126、Transferrin、miR-133b、miR-342和Cystatin C对2型糖尿病肾病的诊断价值

目的探讨miR-126、Transferrin、miR-133b、miR-342和Cystatin C对2型糖尿病肾病的诊断价值。方法选取2型糖尿病肾病患者30例为糖尿病肾病组,2型糖尿病非肾病患者30例为糖尿病非肾病组,同时选取健康体检者30例为对照组,检测3组受试者血清中miR-126和Transferrin的水平,尿中miR-133b、miR-342和Cystatin C的水平。用ROC曲线下面积分析miR-126、Transferrin、miR-133b、miR-342和Cystatin C对2型糖尿病肾病的诊断价值。结果糖尿病肾病患者的miR-126的相对表达量明显低于糖尿病非肾病患者和健康体检者(P<0.05),而其miR-133b和miR-342的相对表达量、Cystatin C和Transferrin的表达量明显高于糖尿病非肾病患者和健康体检者(P <0. 05)。糖尿病非肾病患者miR-126的相对表达量明显低于健康体检者(P <0.05),而其miR-133b的相对表达量和Cystatin C和Transferrin的表达量明显高于健康体检者(P<0.05)。2型糖尿病肾病患者与非2型糖尿病肾病人员的miR-126、miR-133b、miR-342、Cystatin C和Transferrin的灵敏度分别为96%、83%、70%、77%、83%;特异性分别为41%、75%、82%、88%、80%。四项联合检测的灵敏度为97%、特异性为77%,准确率为83%。结论 miR-126、Transferrin、miR-133b、miR-342和Cystatin C可作为诊断2型糖尿病肾病的潜在标志物。

Transferrin receptor 1 plays an important role in muscle development and denervation-induced muscular atrophy

Previous studies demonstrate an accumulation of transferrin and transferrin receptor 1(TfR1) in regenerating peripheral nerves.However, the expression and function of transferrin and TfR1 in the denervated skeletal muscle remain poorly understood.In this study, a mouse model of denervation was produced by complete tear of the left brachial plexus nerve.RNA-sequencing revealed that transferrin expression in the denervated skeletal muscle was upregulated, while TfR1 expression was downregulated.We also investigated the function of TfR1 during development and in adult skeletal muscles in mice with inducible deletion or loss of TfR1.The ablation of TfR1 in skeletal muscle in early development caused severe muscular atrophy and early death.In comparison, deletion of TfR1 in adult skeletal muscles did not affect survival or glucose metabolism, but caused skeletal muscle atrophy and motor functional impairment, similar to the muscular atrophy phenotype observed after denervation.These findings suggest that TfR1 plays an important role in muscle development and denervation-induced muscular atrophy.This study was approved by the Institutional Animal Care and Use Committee of Beijing Institute of Basic Medical Sciences, China(approval No.SYXK 2017-C023) on June 1, 2018.

Transferrin and Its Isoforms from Normal Human Serum Revealed by Several Analytical Techniques

Transferrin（TF） and its isoforms have been widely reported via various analytical techniques, including a noticeable increased number of isoforms with low content of sialic acid（asialo-, monosialo-, and disialo-transferrin） and asialo-TF as well as disialo-TF, with one or several oligosaccharides released in human serum transferrin（hTf）. Here, hTf has been purified by native gradient polyacrylamide gel electrophoresis（PAGEso） before use. The hTf extracted with the electron-transfer approach showed a single subunit band（77.1 Da） in the SDS-PAGE gel, but it exhibited two bands in the native and denatured isoelectric focusing（IEF） gels, namely, hTf-2Fe^3＋ and apo-hTf, without finding any other transferrin isoforms. A reversed phase HPLC（RP-HPLC） equipped with a C18 column effectively separated hTf and its polymers and combined off-line techniques, including peptide mass fingerprinting（PMF）, matrix-assisted laser desorption/ionization time-of-flight mass spectrometry（MALDI-TOF-MS） and database search, and identified the high homology among hTf, apo-hTf, and their isoforms. Moreover, the elution solution consisting of acetonitrile and formic acid could easily denature both hTf and apo-hTf to form various isoforms during separation with HPLC, indicating that chemical factors lead to the formation of various isoforms in transferrin, artificially, during extraction and separation. The authors claimed that only two transferrin isoforms existed in the NHS, namely, hTf-2Fe^3＋ and apo-hTf, which could be employed in biomarkers, to distinguish the healthy population from many disease sufferers, such as, carbohydrate-deficient transferrin（CDT）

Polyethylenimine (PEI)g-comb-poly(ethylene glycol)-transferrin(I): Tumor-targeted Vector for Gene Delivery In-vitro

The work described the synthesis and evaluation of PEI-g-comb-PEG-transferrin as a potential system for gene therapy in vitro. The MW of PEG was 10KDa, and PEI was 2KDa. Its structure was identified by NMR, FT-IR and TGA spectroscopy. MTT assay found that at concentration up to 4000 n mol/L of the polymer, cell viability was over 85%. The bio-character of polymer/DNA complex was characterized by agarose gel electrophoresis, ethidium bromide exclusion and zeta-potential assay. The polymer could retardate DNA at N/P ratio 3.0-3.5 （mol/mol）. The particle size of the polymer/DNA complex was less than 300 nm. Transfection efficiency of the complex was studied in COS7 and NT2 cell lines.

Colorectal cancer screening: Comparison of transferrin and immuno fecal occult blood test

AIM: To evaluate the sensitivity and specificity of transfesrrin dipstick test (Tf) in colorectal cancer (CRC) screening and precancerous lesions screening. METHODS: Eight hundreds and sixty-one individuals at high-risk for CRC were recruited. Six hundreds and eleven subsequently received the three fecal occult blood tests and colonoscopy with biopsy performed as needed. Fecal samples were obtained on the day before colonoscopy. Tf, immuno fecal occult blood test (IFOBT) and guaiac fecal occult blood test (g-FOBT) were performed simultaneously on the same stool. To minimize false-negative cases, all subjects with negative samples were asked to provide an additional stool specimen for a second test even a third test. If the results were all negative after testing three repeated samples, the subject was considered a true negative. The performance characteristics of Tf for detecting CRC and precancerous lesions were examined and compared to those of IFOBT and the combination of Tf, IFOBT and g-FOBT. RESULTS: A total of six hundreds and eleven subjects met the study criteria including 25 with CRC and 60 with precancerous lesions. Sensitivity for detecting CRC was 92% for Tf and 96% for IFOBT, specificities of Tf and IFOBT were both 72.0% (95% CI: 68.2%-75.5%; χ2 = 0.4, P > 0.05); positive likelihood ratios of those were 3.3 (95% CI: 2.8-3.9) and 3.4 (95% CI: 2.9-4.0), respectively. In precancerous lesions, sensitivities for Tf and IFOBT were 50% and 58%, respectively (χ 2 = 0.8, P > 0.05); specificities of Tf and IFOBT were 71.5% (95% CI: 67.6%-75.1%) and 72.2% (95% CI: 68.4%-75.8%); positive likelihood ratios of those were 1.8 (95% CI: 1.3-2.3) and 2.1 (95% CI: 1.6-2.7), respectively; compared to IFOBT, g-FOBT+ Tf+ IFOBT had a significantly higher positive rate for precancerous lesions (83% vs 58%, respectively; χ 2 = 9.1, P < 0.05). In patients with CRC and precancerous lesions, the sensitivities of Tf and IFOBT were 62% and 69% (χ 2 = 0.9, P > 0.05); specificities of those were 74.5% (95% CI: 70.6%-78.1%) and 75.5% (95% CI: 71.6%-79.0%); positive likelihood ratios of those were 2.5 (95% CI: 2.0-3.1) and 2.8 (95% CI: 2.3-3.5). Compared to IF-OBT alone, combining g-FOBT, IFOBT and Tf led to significantly increased sensitivity for detecting CRC and cancerous lesions (69% vs 88%, respectively; χ 2 = 9.0, P < 0.05). CONCLUSION: Tf dipstick test might be used as an ad- ditional tool for CRC and precancerous lesions screening in a high-risk cohort.

Time- and cell-type specific changes in iron, ferritin, and transferrin in the gerbil hippocampal CA1 region after transient forebrain ischemia

In the present study, we used immunohistochemistry and western blot analysis to examine changes in the levels and cellular localization of iron, heavy chain ferritin（ferritin-H）, and transferrin in the gerbil hippocampal CA1 region from 30 minutes to 7 days following transient forebrain ischemia. Relative to sham controls, iron reactivity increased significantly in the stratum pyramidale and stratum oriens at 12 hours following ischemic insult, transiently decreased at 1–2 days and then increased once again within the CA1 region at 4–7 days after ischemia. One day after ischemia, ferritin-H immunoreactivity increased significantly in the stratum pyramidale and decreased at 2 days. At 4–7 days after ischemia, ferritin-H immunoreactivity in the glial components in the CA1 region was significantly increased. Transferrin immunoreactivity was increased significantly in the stratum pyramidale at 12 hours, peaked at 1 day, and then decreased significantly at 2 days after ischemia. Seven days after ischemia, Transferrin immunoreactivity in the glial cells of the stratum oriens and radiatum was significantly increased. Western blot analyses supported these results, demonstrating that compared to sham controls, ferritin H and transferrin protein levels in hippocampal homogenates significantly increased at 1 day after ischemia, peaked at 4 days and then decreased. These results suggest that iron overload-induced oxidative stress is most prominent at 12 hours after ischemia in the stratum pyramidale, suggesting that this time window may be the optimal period for therapeutic intervention to protect neurons from ischemia-induced death.

Preparation and Identification of scFv and bsFv against Transferrin Receptor

To obtain single chain variable fragment （scFv） and bivalent single chain variable fragment （bsFv） against transferrin receptor, up-stream and down-stream primers were designed according to the complementary sequences of FR1 region of variable heavy （VH） and FR4 of variable light （VL）, respectively, which contained inter-linker G4S and the restriction endonuclease SfiI, AscI and NotI. Two pieces of scFv fragments were first amplified through PCR and then inserted into plasmid pAB1, which could express scFv protein once induced by IPTG in the host bacteria. To express scFv and bsFv, E. coli TG1 was cultured in LB broth and was induced by IPTG. The restriction enzyme digestion map and DNA sequencing demonstrated that scFv and bsFv genes were successfully inserted into the expression plasmid. SDS-PAGE and Western blotting revealed the protein band at 35kD and 60kD, which were consistent with the molecular weight of scFv and bsFv respectively. Flow cytometry showed that scFv and bsFv harbored the specific binding activity with TfR expressed in various tumor cells, and the avidity of bsFv was higher than that of the parent scFv.

International Transferring Trend in Manufacturing Industry

This paper addressed the international transferring t endency of manufacturing industry one by one as follows: (1) The developed count ries realize manufacturing industry structure upgrade reling on worldwide tr ansference not only domestic transference. On one hand, the TNCs make cross-inv estments, transnational merger & acquisition so as to achieve the optimum colloc ation of R&D resources, and obtain innovation in knowledge and technology. On th e other hand, the developed countries transfer labor and resource intensive indu stry as well as the labor-intensive link of the production chain in highly tech nology industry to the developing countries, to make their industrial structure escalation. (2) The adjustment and upgrade of manufacturing industry presents "m ulti-step type". First, the object of international transference substitutes th e capital-intensive industry and maturing industry with complicate technique fo r the industry with labor-intensive, simple technique or higher pollution, or s ubstitutes non-core complex technological procedure and parts and components fo r assembly in manufacturing process. Secondly, technology transference forms "br ain-hand and foot" pattern, i.e. the developed countries are increasingly p aying more and more attention to the research and control of core technology, th us transferring simple technique and labor-intensive procedure in higher scienc e and technology. Thirdly, the developed countries begins to transfer both capit al and technology intensive production, as well as transfer certain procedures i n higher technology production process to a few developing countries, even t o transfer parts of R & D to developing countries. (3) TNCs promote the global m anufacturing network (GMN) by recombination and transfer. TNCs organize producti on, trade, technological research and transference through foreign direct invest ment (FDI) all over the world, as a result, GMN makes the national competence no longer depend on absolute occupation of certain industries, but on national com prehensive strength and comparative superiority. TNCs try to race to seize the h igher location of industrial value chain, only leaving the core technology and h igher value-added chains in mother countries, on the other hand, they transfer lower manufacturing power more and more to other countries. (4) TNCs transfer th eir domestic manufacturing bases abroad, China would become world- manufacturin g center. Chinese market is increasingly arousing foreign investor’s interest, " Made in China" will be a synonym of manufacturing industry in new century, subst ituting for "Made in U.S.", "Made in Japanese" and "Made in Europe". The Chinese manufacturing industry should hold tightly the opportunity to melt into the wor ldwide recombination and transfer and enter GMN, so as to accomplish leaping dev elopment.

Studies on the mitogenic effect of transferrin by membrane signal transduction

One of the earliest events leading to cell activation and growth is the hydrolysis of inositol phospholipids producing various membrane signals induced by an interaction between growth factors or hormones with their respective receptors on the cell membrane [1]. To demonstrate the mitogenio action of transferrin, our results show that an addition of transferrin to 'serum-deprived' rat hepatoma cells produced a rapid but transient rise in inositol 1, 4, 5-trisphosphate (IP3) level, and at the same time, an increased intraoellular Oa2+ activity and a oytoplasmic alkalinization were observed. These signal transduo-tions further lend support to the mitogenic nature of transferrin. In addition, a possible link between the receptor-mediated endocytosis of transferrin with the generation of intraoellular signals is discussed herewith.

Novel genetic variants of transferrin receptor 2 exon 4 and cytokines profile of anemic and nonanemic pregnant women in Central Java, Indonesia

Objective:To assess the association between genetic variants of transferrin receptor 2(TFR2)exon 4 and anemia status and to describe the expression levels of several cytokines,hepcidin,soluble transferrin receptor and erythropoietin.Methods:Institutional based comparative study was done randomly to recruit 106 pregnant women who attended antenatal care in three different health centers in Boyolali Regency,Central Java from May 2015 to September 2015.DNA was extracted from peripheral blood samples of selected pregnant women and sequencing was done for TFR2 exon 4.Furthermore,enzyme-linked immunosorbent assay was conducted to measure the expression levels of interleukin 6,interleukin 4,transforming growth factorβand iron-metabolism related proteins such as hepcidin,soluble transferrin receptor,and erythropoietin.Gene alignment was performed by using a CLUSTAL W program.Collected data were analyzed statistically by using parametric and nonparametric tests with Statistical Product and Service Solutions(SPSS)20.0 for Windows.Results:Three novel genetic variants from TFR2 exon 4(position 603,605 and 606)were associated with anemia status.Moreover,the expression levels of interleukin 6,interleukin 4,transforming growth factorβand erythropoietin were higher in anemic pregnant women than those of nonanemic pregnant women but only erythropoietin level reached statistical significance.These results were followed by decreases of hepcidin and soluble transferrin receptor levels.Conclusions:Various factors contribute to anemia prevalence among pregnant women in Boyoali Regency,Central Java,Indonesia.Our novel findings showed that TFR2 exon 4 has 3 mutational sites in position 603,605 and 606.These novel genetic variants may provide a new insight into the role of TFR2 in anemia.

Effects of Acute Temperature Stress on mRNA Expression of Transferrin in the Yellow Pond Turtle Mauremys mutica

The yellow pond turtle Mauremys mutica is widely cultured using both greenhouse-reared and outdoor pond-reared models.Individuals from the two models often show different tolerances to dramatic temperature changes caused by extreme weather events.However,the mechanism underlying the difference is unclear.In this study,we found that for greenhouse-reared turtles(GRTs),the expression levels of an immune-related gene for transferrin were significantly different(P<0.05)between the control group and the acute cold stress(ACS)group for most time points(3 h,6 h and 48 h),while at two time points(6 h and 12 h)there was a significant difference(P<0.05)between the control group and the acute heat stress(AHS)group.However,for the outdoor pond-reared turtles(OPTs),we found the opposite pattern:the ACS group showed no significant difference(P>0.05)from the control group for all time points(3 h,6 h,12 h,24 h and 48 h),whereas two time points(12 h and 24 h)were significantly different(P<0.05)for the AHS group.Our results indicate that ACS may influence the immunity of GRTs and have no influence on OPTs,whereas AHS may largely affect the immunity of OPTs and have little influence on GRTs.The findings provide insights into the mechanism underlying the different morbidity and mortality rates of turtles from different culture models after extreme weather events.

Selection for Anti-transferrin Receptor Bispecific T-cell Engager in Different Molecular Formats

Selecting an ideal molecular format from diverse structures is a major challenge in developing a bispecific antibody(BsAb).To choose an ideal format of anti-CD3 x anti-transferrin receptor(TfR)bispecific antibodies for clinical application,we constructed TfR bispecific T-cell engager(BiTE)in two extensively applied formats,including single-chain tandem singlechain variable fragments(scFvs)and double-chain diabodies,and evaluated their functional characterizations in vitro.Results demonstrated that TfR-BiTE in both formats directed potent killing of TfR+HepG2 cells.However,compared to two・chain diabodies,scFvs were more efficient in antigen binding and TfR target killing.Furthermore,different domain orders in scFvs would also be evaluated because single-TfR-CD3-His was preferable to single-CD3-TfR-His in immunotherapeutic strategies.Thus,the single-chain tandem TfR-CD3 format was favored for further investigation in cancer therapy.