Abstract The clinical use of the single-chain fixed-variable (scFv) fragments of recombinant monoclonal antibodies as credible alternatives for classic therapeutic antibodies has two limitations: rapid blood clearance and inefficient local expression of functional molecules. In attempt to address these issues, we have developed a novel gene therapy protocol in which the anti-death receptor 5 (DR5) scFv fragments were either in vitro expressed in several tumor cell lines, or in vivo expressed in mice, using recombinant adeno-associated virus (rAAV)–mediated gene transfer. Viral transduction using the rAAV-S3C construct, which encodes a scFv molecule (S3C scFv) specific to DR5, led to stable expression in tumor cell lines and showed apoptosis-inducing activity in vitro, which could be inhibited by recombinant DR5 but not by DR4. A single i.m. injection of rAAV-S3C virus in nude mice resulted in stable expression of DR5-binding S3C scFv proteins in mouse sera for at least 240 days. Moreover, the expression of S3C scFv was associated with significant suppression of tumor growth and the increase of tumor cell apoptosis in previously established s.c. human lung LTEP-sml and liver Hep3B tumor xenografts. (Cancer Res 2006; 66(24): 11946-53)
<div>Abstract<p>The clinical use of the single-chain fixed-variable (scFv) fragments of recombinant monoclonal antibodies as credible alternatives for classic therapeutic antibodies has two limitations: rapid blood clearance and inefficient local expression of functional molecules. In attempt to address these issues, we have developed a novel gene therapy protocol in which the anti-death receptor 5 (DR5) scFv fragments were either <i>in vitro</i> expressed in several tumor cell lines, or <i>in vivo</i> expressed in mice, using recombinant adeno-associated virus (rAAV)–mediated gene transfer. Viral transduction using the rAAV-S3C construct, which encodes a scFv molecule (S3C scFv) specific to DR5, led to stable expression in tumor cell lines and showed apoptosis-inducing activity <i>in vitro</i>, which could be inhibited by recombinant DR5 but not by DR4. A single i.m. injection of rAAV-S3C virus in nude mice resulted in stable expression of DR5-binding S3C scFv proteins in mouse sera for at least 240 days. Moreover, the expression of S3C scFv was associated with significant suppression of tumor growth and the increase of tumor cell apoptosis in previously established s.c. human lung LTEP-sml and liver Hep3B tumor xenografts. (Cancer Res 2006; 66(24): 11946-53)</p></div>
Abstract Functional immaturity of neonatal T cells is related to their immature phenotype, with the majority of neonatal T cells of naive (CD45RA+) T cells. The progression of T cells from naive cells to effector cells is dependent on the survival of Ag-specific T cells and their resistance to apoptosis. In this study, we showed for the first time that insulin-like growth factor 1 (IGF-1) converted cord blood CD45RA+ T cells to CD45RO+ T cells and inhibited cord blood T cell apoptosis. We found cord blood T cells stimulated with PHA would result in gradual loss of CD45RA and gain of CD45RO expression. IGF-1 further increased the loss of CD45RA and enhanced CD45RO expression in PHA-stimulated cord blood T cells. In addition, IGF-1 prevented cord blood T cells from spontaneous apoptosis through a mechanism other than Fas/FasL. In PHA-activated cord blood T cells, IGF-1 prevented both naive (CD45RA+) and memory/mature (CD45RO+) T cells from apoptosis. Moreover, cord blood T cells cultured with IGF-1 and PHA had a higher resistance to anti-Fas-induced apoptosis as compared with PHA-activated cord blood T cells. IGF-1 also significantly inhibited PHA-induced Fas expression on cord blood T cells. These results demonstrate that IGF-1 promotes the maturation and maintains the survival of cord blood T cells. Its antiapoptotic effect in PHA-activated cord blood T cells may be mediated through the down-regulation of Fas expression.
To investigate the effect of estrogen on cell proliferation and expression of proteins of C-type natriuretic peptide (CNP), natriuretic peptides B receptor (NPR-B) and natriuretic peptides C receptor (NPR-C) in ATDC5 cells during chondrogenesis.ATDC5 cells were induced for differentiation with insulin 10 µg/ml (day 0), and were started to be investigated on day 6. They were incubated with: (1) Estradiol (E2) at different concentrations (10(-11)-10(-5) mol/L) for 24 hours (for studying cell proliferation), or for 48 hours (for studying CNP, NPR-B and NPR-C protein expression); (2) E2 (10(-8) mol/L) for 24, 48, 72, 96 and 120 h (for studying cell proliferation), or for 24, 48, 72 and 96 hours (for studying CNP, NPR-B and NPR-C protein expression); (3) E2 (10(-8) mol/L) , and/or ICI 182782 (estrogen receptor antagonist ) (10(-7) mol/L) for 24 hours (for studying cell proliferation). ATDC5 cells proliferation were determined by MTT (OD value). Western-blotting was performed to identify the protein levels of CNP, NPR-B and NPR-C.(1) After incubation with E2 (10(-11)-10(-5) mol/L) for 24 h, ATD5 cell number increased with the increasing E2 concentration, peak in E2 concentrations of 10(-9) and 10(-8) mol/L (0.56 ± 0.06 and 0.52 ± 0.02, P < 0.05 and <0.01, respectively) , while significantly decreased in E2 (10(-5) mol/L) (0.30 ± 0.02) compared with DMSO-control (0.38 ± 0.02) (P < 0.05). After incubation with E2 (10(-11)-10(-5) mol/L) for 48 h, the protein level of CNP, NPR-B and NPR-C increased significantly, with the greatest effect seen at a concentration of 10(-10) mol/L E2 for CNP and NPR-B, 10(-9) mol/L E2 for NPR-C (P < 0.05). (2) After incubation with E2 (10(-8) mol/L) for 24 to 96 hours: (1) The cell number in each of the four time points was significantly increased compared with DMSO-control, with the greatest effect in 48 h (0.030 ± 0.003) (P < 0.05 or <0.01, respectively). While the cell number at 120 h was similar to that in DMSO-control. (2) The protein level of CNP increased significantly at 24 h (P < 0.05), seemed to be increased at 48 h and 72 h and decreased at 96 h. Both NPR-B and NPR-C level seemed to be increased at 24 h (P = 0.060 and 0.055, respectively) and seemed to decrease at 48 h, with decreasing significantly at both 72 h and 96 h (P < 0.05). (3) After incubation for 24 h, there was significant difference among the cell number of the four groups (P < 0.05). Cell number of group E2 (0.470 ± 0.032) was increased compared with group (E2+ICI) (0.410 ± 0.018), both being increased compared with group DMSO-control (0.370 ± 0.011, P < 0.05, respectively). There was no difference in cell number between group ICI 182782(0.360 ± 0.035) and group DMSO-control.E2 promotes the proliferation of ATDC5 cells i.e. chondrogenesis via estrogen receptor mediated mechanism, in both concentration-dependent and time-dependent manner. E2 (10(-11)-10(-8) mol/L) up-regulates protein expression of CNP, NPR-B and NPR-C of ATDC5 cells during chondrogenesis, and regulate the expression of the three proteins mentioned above positively or negatively at different time point, which implied that estrogen is one of the regulators of CNP signaling pathway.
Background: The outbreak of Coronavirus Disease 2019 (COVID-19) caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has wildly spread in Europe and the United States. Local human-to-human-transmission indicates the strong infectivity of SARS-CoV-2 and about 80% of the cluster transmission of COVID-19 occurred in families in China. Children as special population are susceptible to SARS-COV-2 at all ages. The epidemiological characteristics and non-pharmacological interventions of pediatric COVID-19 patients in Hubei by 5 periods according to key events and interventions were discussed.Methods: A retrospective study was conducted on the epidemiological characteristics of pediatric COVID-19 patients in Hubei Province from January to May,2020, including patients' age, sex, residential location, severity classification, date of onset of symptoms and date of diagnosed.Findings: By 24:00 hours on April 21th, 2020, a total number of 68128 confirmed cases were registered, of which 1304 were pediatric confirmed cases, median age was 9.01 years old (intertitles range:3.43-13.84 years). 756 (57.98%) were male and 548 (42.02%) were female. The number of cases account for 2.68%, 8.67%, 23.85%, 25.08%, 27.07%, 12.65% in neonates, infants, 1-5 years old, 6-10 years old, 11-15 years old and >15 years old respectively.226 (17.33%) were asymptomatic, 637 (48.85%) were mild cases and 396 (30.37%) were moderate cases. There were 42 severe cases and 3 critical cases, accounting for 3.22% and 0.23%.The total rate of confirmed cases is 534 per million in Wuhan and 118 per million in non-Wuhan in children. Thirty patients of severe and critical COVID-19 cases have onset of symptoms before February 12th, 2020 . The average daily rate of pediatric confirmed cases of children in Hubei Province is 0.67 per million (first phase), 2.90 per million (second phase), 4.28 per million (third phase), 0.25 per million (fourth phase),0.04 per million (fifth phase) and 0.03 per million (sixth phase).Statistically significant difference was found in different epidemic phases with a sharp decrease in the fourth phase. Interpretation: The proportion of pediatric COVID-19 is lower than that of adults and people in all ages are susceptible including newborns. Pediatric COVID-19 cases are less severe and the rate of confirmed cases are much lower than those in adults in Hubei. There are four keys points to control the epidemic outbreak quickly in children: 1) The centralized quarantine of patients, which protects children from being infected; 2) The community and city lockdown and the limit of social activity, which protects the elder children; 3) The public intervention, such as wearing face mask and hand hygiene, which protects the infants; 4) The compulsory monitored body temperature to investigate and isolate the potential patients in time. Funding: 1.Tongji hospital of Tongji medical college, Huazhong University of Science and Technology No.XXGZBDYJ0052.Major Special Science and Technology Project of Hubei Province under Grant No. 2020AEA0093.Application foundation frontier project of Wuhan No.2020020601012228Declaration of Interest: All the authors, including Liru Qiu, Yue Zhao, Wenhua Liu, Xi Sun, Pik-to Cheung, Sainan Shu, Yan Hao, Yu Chen, Yan Liu, Menaka Dhuromsingh,Jianhua Zhou, Feng Fang,Qin Ning and Xiaoping Luo declare no competing interests.Ethical Approval: All data analysis in this case were approved by the Ethical Committee of TongjiHospital of Huazhong University of Science & Technology (China) (TJC20200359)
Type 1 diabetes (T1D) incidence varies substantially between countries/ territories, with most studies indicating increasing incidence. In Western Pacific region (WPR), reported rates are much lower than European-origin populations. In contrast, there are reports of substantial numbers of young people with type 2 diabetes (T2D). A deeper understanding of T1D and T2D in the WPR may illuminate factors important in pathogenesis of these conditions. Furthermore, with varying resources and funding for diabetes treatment in this region, there is a need to more clearly determine the current burden of disease and also any gaps in knowledge.
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