Exposure to user-generated e-cigarette content on social media associated with greater vulnerability to e-cigarette use among youth non-users.

INTRODUCTION: Social media are important venues for youth's exposure to e-cigarette content. This study examined how exposure to user-generated e-cigarette content (i.e., content created and shared by individual social media users) is associated with vulnerabilities to e-cigarette use among youth non-users. METHODS: We pooled data from the 2021 and 2022 National Youth Tobacco Survey. Youth who have never used e-cigarettes were included. Weighted linear and logistic regressions were conducted to examine how exposure to user-generated e-cigarette content (from real-life friends, online-only friends, and celebrities/influencers) on social media was associated with e-cigarette use vulnerabilities measured by perceived norms, perceived risk, and susceptibility of use, controlling for demographics, advertising exposure, and mental health conditions. Multiple imputations were performed to account for missing data. RESULTS: Exposure to e-cigarette content on social media posted by real-life friends, online-only friends, and celebrities/influencers were associated with more positive descriptive norm (ßs = 1.56, 0.37, and 0.35, respectively, all ps < .001), more positive injunctive norm (ßs = 0.46, 0.19, and 0.10, respectively, all ps < .001), and higher odds of e-cigarette use susceptibility (ORs = 1.48, 1.50. 1.29, respectively, all ps < .001). Exposure to content posted by real-life and online-only friends were associated with reduced risk perception of e-cigarette use (ß = -0.04, p < 0.05 and ß = -0.07, p < 0.001). CONCLUSIONS: Our study highlighted that friends and celebrities/influencers are important sources on social media that can influence youth non-users' vulnerabilities to e-cigarette use. Interventional messages communicated through friends and influencers on social media may in turn help reduce e-cigarette vulnerability among youth non-users.

Important role and underlying mechanism of non­SMC condensin I complex subunit G in tumours (Review).

At present, the incidence of tumours is increasing on a yearly basis, and tumourigenesis is usually associated with chromosomal instability and cell cycle dysregulation. Moreover, abnormalities in the chromosomal structure often lead to DNA damage, further exacerbating gene mutations and chromosomal rearrangements. However, the non­SMC condensin I complex subunit G (NCAPG) of the structural maintenance of chromosomes family is known to exert a key role in tumour development. It has been shown that high expression of NCAPG is closely associated with tumour development and progression. Overexpression of NCAPG variously affects chromosome condensation and segregation during cell mitosis, influences cell cycle regulation, promotes tumour cell proliferation and invasion, and inhibits apoptosis. In addition, NCAPG has been associated with tumour cell stemness, tumour resistance and recurrence. The aim of the present review was to explore the underlying mechanisms of NCAPG during tumour development, with a view towards providing novel targets and strategies for tumour therapy, and through the elucidation of the mechanisms involved, to lay the foundation for future developments in health.

The Value of Dual-Energy Computed Tomography-Based Radiomics in the Evaluation of Interstitial Fibers of Clear Cell Renal Carcinoma.

OBJECTIVE: We investigated the potential of dual-energy computed tomography (DECT) radiomics in assessing cancer-associated fibroblasts in clear cell renal carcinoma (ccRCC). METHODS: A retrospective analysis was conducted on 132 patients with ccRCC. The arterial and venous phase iodine-based material decomposition images (IMDIs), virtual non-contrast images, 70 keV, 100 keV, and 150 keV virtual monoenergetic images, and mixed energy images (MEIs) were obtained from the DECT datasets. On the Radcloud platform, radiomics feature extraction, feature selection, and model establishment were performed. Seven radiomics models were established using the support vector machine. The predictive performance was evaluated by utilizing receiver operating characteristic and the area under the curve (AUC) was calculated. Nomograms were constructed. RESULTS: The combined model demonstrated high efficiency in evaluating pseudocapsule thickness with AUC, specificity, and sensitivity of 0.833, 0.870, and 0.750, respectively in the validation set, surpassing those of other models. The precision, F1-score, and Youden index were also higher for the combined model. For evaluating the number of collagen fibers, the combined model exhibited the highest AUC (0.741) among all models, with a specificity of 0.830 and a sensitivity of 0.330. The AUC in the 150 kv model and IMDI model were slightly lower than those in the combined model (0.728 and 0.710, respectively), with corresponding sensitivity and specificity of 0.560/0.780 and 0.670/0.830. The nomogram exhibited that Rad-score had good prediction efficiency. CONCLUSION: DECT radiomics features have significant value in evaluating the interstitial fibers of ccRCC. The combined model of IMDI + MEI exhibits superior performance in assessing the thickness of the pseudocapsule, while the combined, 150 keV, and IMDI models demonstrate higher efficacy in evaluating collagen fiber number. Radiomics, combined with imaging features and clinical features, has excellent predictive performance. These findings offer crucial support for the clinical diagnosis, treatment, and prognosis of ccRCC and provide valuable insights into the application of DECT.

TaCRT3 Is a Positive Regulator of Resistance to Blumeria graminis f. sp. tritici in Wheat.

Wheat powdery mildew, caused by Blumeria graminis f. sp. tritici (Bgt), is one of the most prevalent diseases of wheat worldwide and can lead to severe yield reductions. Identifying genes involved in powdery mildew resistance will be useful for disease resistance breeding and control. Calreticulin (CRT) is a member of multigene family widely found in higher plants and is associated with a variety of plant physiological functions and defense responses. However, the role of CRT in wheat resistance to powdery mildew remains unclear. TaCRT3 was identified from the proteomic sequence of an incompatible interaction between the wheat (Triticum aestivum) cultivar Xingmin 318 and the Bgt isolate E09. Following analysis of transient expression of the GFP-TaCRT3 fusion protein in Nicotiana benthamiana leaves, TaCRT3 was localized in the nucleus, cytoplasm, and cell membrane. Transcript expression levels of TaCRT3 were significantly upregulated in the wheat-Bgt incompatible interaction. More critically, knockdown of TaCRT3 using virus-induced gene silencing resulted in attenuated resistance to Bgt in wheat. Histological analysis showed a significant increase in Bgt development in TaCRT3-silenced plants, whereas the pathogen-related gene was significantly downregulated in TaCRT3-silenced leaves. In addition, overexpression of TaCRT3 in wheat enhanced the resistance to powdery mildew, the growth of Bgt was significantly inhibited, and the area of H2O2 near the infection site and the expression of defense-related genes of the salicylic acid pathway significantly increased. These findings imply that TaCRT3 may act as a disease resistance factor that positively regulates resistance to powdery mildew, during which SA signaling is probably activated.

CEAC (oral semustine, etoposide, cytarabine and cyclophosphamide) vs BEAM (carmustine, etoposide, cytarabine, and melphalan) conditioning regimen of autologous stem cell transplantation for diffuse large B-cell lymphoma: a post-hoc, propensity score-matched, cohort study in Chinese patients.

Autologous stem cell transplantation (ASCT) is a salvage therapy for relapsed or refractory diffuse large B-cell lymphoma (DLBCL). We have developed a novel conditioning regimen called CEAC (oral semustine 250 mg/m2 d-6, etoposide 300 mg/m2 d-5 ~ d-2, cytarabine 500 mg/m2 d-5 ~ d-2, and cyclophosphamide 1200 mg/m2 d-5 ~ d-2) In lymphoma patients in China. Here, we conducted a study to compare the conventional BEAM regimen with the CEAC regimen in 110 DLBCL patients. Propensity-score matching was performed in a 1:4 ratio (22 patients received BEAM and 88 received CEAC). Our results showed no significant difference in the overall response rate (95% vs 97%, P = 1.000) and complete response rate (66% vs 73%, P = 0.580) between the two cohorts. The 5-year progression-free survival (PFS), 5-year overall survival (OS), and 5-year cumulative incidence of relapse (CIR) for all patients were 72% (95% CI 62%-82%), 92% (95% CI 86%-97%), and 29% (95% CI 17%-38%), respectively. There was no significant difference in the 5-year PFS (80% vs 70%, P = 0.637), 5-year OS (95% vs 91%, P = 0.496), and 5-year CIR (20% vs 30%, P = 0.733) between cohorts. In terms of safety, the CEAC cohort had a lower incidence rate of grade 1-2 gastrointestinal hemorrhage (P = 0.023) and severe nausea (P = 0.007) compared with the BEAM cohort. In conclusion, the CEAC regimen seems to be a suitable alternative to the BEAM regimen for ASCT in DLBCL patients.

The value of a dual-energy CT Iodine map radiomics model for the prediction of collagen fiber content in the ccRCC tumor microenvironment.

BACKGROUND AND PURPOSE: Renal cell carcinoma (RCC) is a heterogeneous group of cancers. The collagen fiber content in the tumor microenvironment of renal cancer has an important role in tumor progression and prognosis. A radiomics model was developed from dual-energy CT iodine maps to assess collagen fiber content in the tumor microenvironment of ccRCC. METHODS: A total of 87 patients with ccRCC admitted to our hospital were included in this retrospective study. Among them, 59 cases contained large amounts of collagen fibers and 28 cases contained a small amount of collagen fibers. We established a radiomics model using preoperative dual-energy CT scan Iodine map (IV) imaging to distinguish patients with multiple collagen fibers from those with few collagen fibers in the tumor microenvironment of ccRCC. We extracted features from dual-energy CT Iodine map images to evaluate the effects of six classifiers, namely k-nearest neighbor (KNN), support vector machine (SVM), extreme gradient boosting (XGBoost), random forest (RF), logistic regression (LR), and decision tree (DT). The effects of the models built based on the dynamic and venous phases are also compared. Model performance was evaluated using quintuple cross-validation and area under the receiver operating characteristic curve (AUC). In addition, a clinical model was developed to assess the clinical factors affecting collagen fiber content. RESULTS: Compared to KNN, SVM, and LR classifiers, RF, DT, and XGBoost classifiers trained with higher AUC values, with training sets of 0.997, 1.0, and 1.0, respectively. In the validation set, the highest AUC was found in the SVM classifier with a size of 0.722. In the comparative test of the active and intravenous phase models, the SVM classifier had the best effect with its validation set AUC of 0.698 and 0.741. In addition, there was a statistically significant effect of patient age and maximum tumor diameter on the collagen fiber content in the tumor microenvironment of kidney cancer. CONCLUSION: Radionics features based on preoperative dual-energy CT IV can be used to predict the amount of collagen fibers in the tumor microenvironment of renal cancer. This study better informs clinical prognosis and patient management. Iodograms may add additional value to dual-energy CTs.

Statistical considerations in long-term efficacy evaluation of anti-cancer therapies.

Anti-cancer therapy has been a significant focus of research. Developing and marketing various types and mechanisms of anti-cancer therapies benefit a variety of patients significantly. The long-term benefit to patients in evaluating the risk-benefit ratio of anti-cancer therapy has become a significant concern. This paper discusses the evaluation of long-term efficacy within the estimand framework and summarizes the various strategies for addressing potential intercurrent events. Non-proportional hazards of survival data may arise with novel anti-cancer therapies, leading to potential bias in conventional evaluation methods. This paper reviews statistical methods for addressing this issue, including novel endpoints, hypothesis testing, and efficacy estimation methods. We also discuss the influences of treatment switching. Although advanced methods have been developed to address the non-proportional hazard, they still have limitations that require continued collaborative efforts to resolve issues.

NCAPG is transcriptionally regulated by CBX3 and activates the Wnt/ß-catenin signaling pathway to promote proliferation and the cell cycle and inhibit apoptosis in colorectal cancer.

Background: Colorectal cancer (CRC) is highly heterogeneous at the genetic and molecular level and a major contributor to cancer-death worldwide. Non-structural maintenance of chromosomes (SMC) condensin I complex subunit G (NCAPG) is a subunit of condensin I and has been shown to be associated with the prognosis of cancers. This study investigated the functional role of NCAPG in CRC and its mechanism. Methods: Messenger RNA (mRNA) and protein expressions of NCAPG and chromobox protein homolog 3 (CBX3) were determined by reverse transcription-quantitative polymerase chain reaction (RT-qPCR) and western blot. The proliferation, cycle, and apoptosis of HCT116 cells were analyzed by Cell Counting Kit-8 (CCK-8), flow cytometry, and terminal deoxynucleotidyl transferase dUTP nick-end labeling (TUNEL) assay. RT-qPCR and western blot were used to determine the transfection efficacy of short hairpin (sh)-NCAPG and sh-CBX3. Western blot was used to explore cycle-, apoptosis-, and Wnt/ß-catenin signaling-related proteins, and the activity of NCAPG promoter was evaluated using a luciferase report assay. The expressions of cleaved caspase9 and cleaved caspase3 were assessed by colorimetric caspase activity assay. Results: The results showed that NCAPG expression was elevated in CRC cells. After transfection with sh-NCAPG, NCAPG expression was reduced. It was also discovered that NCAPG knockdown suppressed proliferation and the cell cycle but induced apoptosis in HCT116 cells. The Human Transcription Factor Database (HumanTFDB; http://bioinfo.life.hust.edu.cn/HumanTFDB#!/) predicted the binding sites of CBX3 and NCAPG promoters. Meanwhile, the Encyclopedia of RNA Interactomes (ENCORI) database (https://starbase.sysu.edu.cn/) revealed that CBX3 was positively correlated with NCAPG. Our results showed that NCAPG was transcriptionally regulated by CBX3. Additionally, Wnt/ß-catenin signaling was discovered to be activated by CBX3 overexpression. Further experiments showed that NCAPG transcriptionally regulated by CBX3 activated Wnt/ß-catenin signaling to regulate the proliferation, cell cycle, and apoptosis of HCT116 cells. Conclusions: Collectively, the results of our study indicated that NCAPG was transcriptionally regulated by CBX3 and activated the Wnt/ß-catenin signaling pathway to facilitate the progression of CRC.

Lactate and Lactylation in the Brain: Current Progress and Perspectives.

As the final product of glycolysis, lactate features not only as an energy substrate, a metabolite, and a signaling molecule in a variety of diseases-such as cancer, inflammation, and sepsis-but also as a regulator of protein lactylation; this is a newly proposed epigenetic modification that is considered to be crucial for energy metabolism and signaling in brain tissues under both physiological and pathological conditions. In this review, evidence on lactylation from studies on lactate metabolism and disease has been summarized, revealing the function of lactate and its receptors in the regulation of brain function and summarizing the levels of lactylation expression in various brain diseases. Finally, the function of lactate and lactylation in the brain and the potential mechanisms of intervention in brain diseases are presented and discussed, providing optimal perspectives for future research on the role of lactylation in the brain.

Effect of riboflavin and carbon black co-modified fillers coupled with alkaline pretreatment on anaerobic digestion of waste activated sludge.

Additional various carbon and free riboflavin could improve anaerobic digestion of waste activated sludge (WAS). However, these substances were not reused. In this study, a reusable riboflavin and carbon black (RCB) co-modified filler was developed and combined with alkaline pretreatment for enhancing the production of volatile fatty acids (VFAs) and methane during anaerobic digestion of WAS. The results showed that RCB-modified fillers exhibited a promoting effect on the reduction of alkali-pretreated WAS. The amounts of the accumulated VFAs mainly containing acetate and the produced methane rose with the increased concentration of immobilized riboflavin (0-0.75 g/L) in the presence of 4 g/L carbon black. When the alkaline pretreatment time of WAS increased from 3 d to 8 d, the amount of methane production increased from 22.8% to 63.9% in the presence of 0.75 g/L riboflavin and 4 g/L carbon black compared with that without RCB-modified fillers. Moreover, 0.75 g/L riboflavin and 4 g/L carbon black had a synergetic effect on promoting methane production via broadening extracellular electron transfer pathways. During this process, microbial dehydrogenase activity, electron transport system activity and coenzyme F420 were enhanced. Microbial community analysis showed that RCB-modified filler addition promoted the enrichment of Syntrophomonas and Pseudomonas involved in direct interspecies electron transfer (DIET). These results indicated that DIET establishment was accelerated. Meanwhile, the populations of acetic acid-producing bacteria including Rikenellaceae_RC9_gut_group and Proteiniphilum, aceticlastic and acid-tolerant methanogenic archaea including Methanosarcina and Methanosaeta, RumEn_M2 were increased. These results indicate that RCB-modified fillers coupled with alkaline pretreatment is an effective method to promote the production of methane during anaerobic digestion of WAS.

Wheat Class III Peroxidase TaPOD70 Is a Potential Susceptibility Factor Negatively Regulating Wheat Resistance to Blumeria graminis f. sp. tritici.

Powdery mildew, caused by Blumeria graminis f. sp. tritici (Bgt), is one of the most important diseases on wheat worldwide and can lead to a large reduction in wheat production. Class III peroxidases (PODs), a kind of secretory enzyme and members of a multigene family in higher plants, have been linked to various plant physiological functions and defensive responses. However, the role of PODs in wheat resistance to Bgt remains unclear. TaPOD70, a class III POD gene, was identified from the proteomics sequencing of the incompatible interaction between wheat (Triticum aestivum) cultivar Xingmin 318 and Bgt isolate E09. After transient expression of the TaPOD70-GFP fusion protein in Nicotiana benthamiana leaves, TaPOD70 was located in the membrane region. Yeast secretion assay showed that TaPOD70 was a secretory protein. Furthermore, Bax-induced programmed cell death was inhibited by transient expression of TaPOD70 in N. benthamiana. The transcript expression level of TaPOD70 was significantly upregulated in the wheat-Bgt compatible interaction. More crucially, knocking down TaPOD70 using virus-induced gene silencing increased wheat resistance to Bgt compared with the control plants. In response to Bgt, histological analyses indicated that hyphal development of Bgt was significantly reduced, whereas H2O2 production was enhanced in TaPOD70-silenced leaves. These findings imply that TaPOD70 may act as a susceptibility factor, adversely regulating wheat resistance to Bgt.

A comparative study of Bayesian optimal interval (BOIN) design with interval 3+3 (i3+3) design for phase I oncology dose-finding trials.

Bayesian optimal interval (BOIN) design is a model-assisted phase I dose-finding design to find the maximum tolerated dose (MTD). The hallmark of the BOIN design is its concise decision rule - making the decision of dose escalation and de-escalation by simply comparing the observed dose-limiting toxicity (DLT) rate at the current dose with a pair of optimal dose escalation and de-escalation boundaries. The interval 3+3 (i3+3) design is a recently proposed algorithm-based dose-finding design based on a similar decision rule with some modifications. The similarity in the appearance of the two designs has caused confusions among practitioners. In this article, we demystify the i3+3 design by elucidating its links with the BOIN design and compare their similarities and differences, as well as pros and cons. We perform comprehensive simulation studies to compare the operating characteristics of the two designs. Our results show that, compared to the algorithm-based i3+3 design, which are characterized by ad hoc and often scientifically and logically incoherent decision rules, the mode-assisted BOIN design is not only simpler, but also statistically more rigorous with better operating characteristics, thus providing a better design choice for phase I oncology trials.

Shotgun: A Bayesian seamless phase I-II design to accelerate the development of targeted therapies and immunotherapy.

Drug development of novel antitumor agents is conventionally divided by phase and cancer indication. With the advent of new molecularly targeted therapies and immunotherapies, this approach has become inefficient and dysfunctional. We propose a Bayesian seamless phase I-II "shotgun" design to evaluate the safety and antitumor efficacy of a new drug in multiple cancer indications simultaneously. "Shotgun" is used to describe the design feature that the trial begins with an all-comer dose finding phase to identify the maximum tolerated dose (MTD) or recommended phase II dose (RP2D), and then is seamlessly split to multiple indication-specific cohort expansions. Patients treated during dose finding are rolled over to the cohort expansion for more efficient evaluation of efficacy, while patients enrolled in cohort expansion contribute to the continuous learning of the safety and tolerability of the new drug. During cohort expansion, interim analyses are performed to discontinue ineffective or unsafe expansion cohorts early. To improve the efficiency of such interim analyses, we propose a clustered Bayesian hierarchical model (CBHM) to adaptively borrow information across indications. A simulation study shows that compared to conventional approaches and the standard Bayesian hierarchical model, the shotgun design has substantially higher probabilities to discover indications that are responsive to the treatment in question, and is associated with a reasonable false discovery rate. The shotgun provides a phase I-II trial design for accelerating drug development and to build a more robust foundation for subsequent phase III trials. The proposed CBHM methodology also provides an efficient design for basket trials.

Dietary grape seed procyanidin extract protects against lead-induced heart injury in rats involving endoplasmic reticulum stress inhibition and AKT activation.

To investigate the protective role of grape seed procyanidin extract (GSPE) against lead-induced heart injury and the possible molecular mechanism associated with this event, Wistar rats were orally given GSPE (200 mg/kg) daily with or without lead acetate (PbA) (0.5 g/L) in drinking water for 56 d. GSPE attenuated oxidative stress, heart dysfunction, and lead accumulation in lead-exposed rat hearts. Meanwhile, GSPE inhibited the protein kinase RNA-like endoplasmic reticulum (ER) kinase/eukaryotic initiation factor 2α signaling pathway, and promoted protein kinase B (AKT) and glycogen synthase kinase 3ß phosphorylation altered by lead, and regulated lead-activated apoptosis and its related signaling pathway. This study suggests that dietary GSPE ameliorates lead-induced heart injury associated with ER stress inhibition and AKT activation. Dietary GSPE may be a protector against lead-induced heart injury and a novel therapy for lead exposure.

Sarsaparilla (Smilax Glabra Rhizome) Extract Activates Redox-Dependent ATM/ATR Pathway to Inhibit Cancer Cell Growth by S Phase Arrest, Apoptosis, and Autophagy.

Sarsaparilla (Smilax Glabra Rhizome) exerts growth inhibitory effect on multiple cancer cells in vitro and in vivo, and redox-dependent persistent activation of ERK1/2 has been reported to underlie this effect. Here, we report an activation of ATM/ATR-dependent signaling pathway also as a mechanism for the cancer cell growth inhibition induced by the supernatant fraction of the water-soluble extract from sarsaparilla (SW). SW treatment (3.5 µg/µL) promoted the phosphorylations of ATM, ATR, and CHK1 in AGS and HT-29 cells. The ATM kinase inhibitor, KU55933, could reverse SW-induced ERK phosphorylation but not the reduced glutathione/oxidized glutathione (GSH/GSSG) imbalance in AGS cells. However, both the redox inhibitor glutathione (GSH) and ERK inhibitor U0126 antagonized SW-induced phosphorylations of ATM, ATR, and CHK1 in AGS cells. We further found KU55933 significantly antagonized SW-induced S phase arrest, apoptosis, autophagy and the resultant cell growth inhibition. Our results provide another molecular basis for the anticancer action of sarsaparilla.

Threat Appeals: The Fear-Persuasion Relationship is Linear and Curvilinear.

Drive theory may be seen as the first scientific theory of health and risk communication. However, its prediction of a curvilinear association between fear and persuasion is generally held to be incorrect. A close rereading of Hovland et al. reveals that within- and between-persons processes were conflated. Using a message that advocated obtaining a screening for colonoscopy, this study (N = 259) tested both forms of the inverted-U hypothesis. In the between-persons data, analyses revealed a linear effect that was consistent with earlier investigations. However, the data showed an inverted-U relationship in within-persons data. Hence, the relationship between fear and persuasion is linear or curvilinear depending on the level of analysis.

Enhanced engagement of CTLA-4 induces antigen-specific CD4+CD25+Foxp3+ and CD4+CD25- TGF-beta 1+ adaptive regulatory T cells.

CTLA-4 is a critical negative regulator of T cell response and is instrumental in maintaining immunological tolerance. In this article, we report that enhanced selective engagement of CTLA-4 on T cells by Ag-presenting dendritic cells resulted in the induction of Ag-specific CD4(+)CD25(+)Foxp3(+) and CD4(+)CD25(-)TGF-beta1(+) adaptive Tregs. These cells were CD62L(low) and hyporesponsive to stimulation with cognate Ag but demonstrated a superior ability to suppress Ag-specific effector T cell response compared with their CD62L(high) counterparts. Importantly, treatment of mice with autoimmune thyroiditis using mouse thyroglobulin (mTg)-pulsed anti-CTLA-4 agonistic Ab-coated DCs, which results in a dominant engagement of CTLA-4 upon self-Ag presentation, not only suppressed thyroiditis but also prevented reemergence of the disease upon rechallenge with mTg. Further, the disease suppression was associated with significantly reduced mTg-specific T cell and Ab responses. Collectively, our results showed an important role for selective CTLA-4 signaling in the induction of adaptive Tregs and suggested that approaches that allow dominant CTLA-4 engagement concomitant with Ag-specific TCR ligation can be used for targeted therapy.

Bone marrow is a preferential homing site for autoreactive T-cells in type 1 diabetes.

OBJECTIVE: The pancreatic microenvironment is considered to be the primary location of autoreactive T-cells in type 1 diabetes. Diabetogenic T-cells have also been detected in the spleens of NOD mice. However, it is not known whether bone marrow also contains T-cells specific for self-antigens in hosts with autoimmunity. In this study, we investigated whether autoreactive diabetogenic T-cells are present in the bone marrow of NOD mice. RESEARCH DESIGN AND METHODS: Bone marrow and splenic T-cells of female NOD mice were purified and tested for their cytokine secretion and proliferation in response to stimulation with immunodominant peptides of pancreatic beta-cells. The diabetogenic nature and homing properties of purified bone marrow T-cells were compared with those of splenic T-cells in NOD-Scid and wild-type mice. RESULTS: The bone marrow T-cells from both hyperglycemic and young euglycemic mice demonstrated profoundly higher proliferation and cytokine production in response to stimulation with beta-cell antigens than T-cells from spleen. Bone marrow T-cells showed rapid expansion and aggressive infiltration into pancreatic islets in NOD-Scid mice and induced hyperglycemia earlier than splenic T-cells. Adoptive transfer of bone marrow T-cells resulted in their trafficking predominantly to bone marrow and pancreatic lymph nodes. CONCLUSIONS: Our study demonstrates that a large number of diabetogenic T-cells are present in the bone marrow of female NOD mice and that these autoreactive T-cells can be detected long before clinical onset of the disease.

Dendritic cells from chronic hepatitis B patients can induce HBV antigen-specific T cell responses.

AIM: To determine whether dendritic cells (DCs) from chronic hepatitis B patients could induce HBV antigen-specific T cell responses or not. METHODS: DCs were generated from peripheral blood mononuclear cells of patients with chronic hepatitis B (CHB) infection and healthy donors. We compared the phenotypes of these DCs and their ability to secrete cytokines and to participate in mixed lymphocyte reactions. In addition, autologous lymphocytes were cultured with DCs loaded with HBV core region peptide HBcAg8-27, an epitope recognized by cytotoxic T lymphocytes (CTL), and bearing human leucocyte antigen (HLA)-A2 for 10 d. Cytokine secretion and lytic activity against peptide-pulsed target cells were assessed. RESULTS: DCs with typical morphology were generated successfully by culturing peripheral blood mononuclear cells (PBMCs) from CHB patients with AIM-V containing GM-CSF and IL-4. Compared with DCs from normal donors, the level of CD80 expressed in DCs from CHB patients was lower, and DCs from patients had lower capacity of stimulate T cell proliferation. When PBMCs isolated from patients with chronic or acute hepatitis B infection and from normal donors were cocultured with HBcAg18-27 peptide, the antigen-specific memory response of PBMCs from acute hepatitis B patients was stronger than that of PBMCs from chronic hepatitis B patients or normal donors. PBMCs cocultured with DCs treated with HBcAg18-27 CTL epitope peptide induced an antigen-specific T cell reaction, in which the level of secreted cytokines and lytic activity were higher than those produced by memory T cells. CONCLUSION: DCs from patients with CHB can induce HBV antigen-specific T cell reactions, including secretion of cytokines essential for HBV clearance and for killing cells infected with HBV.

[Functions of cultured dendritic cells from patients with chronic hepatitis Beta decreased].

OBJECTIVE: To compare the phenotype and function of dendritic cells (DCs) of patients with chronic hepatitis Beta with those of DCs from healthy persons. METHOD: Peripheral blood monocytes (PBMCs) were isolated from 10 patients with chronic hepatitis Beta and 10 healthy blood donors. DCs were generated by culturing PBMCs in 1 000 U/ml granulocyte-macrophage stimulating factor (GM-CSF) and 500 U/ml IL-4 with AIM-V in vitro. DCs and supernatant were collected 3, 5, 7, 9, 11, 13 and 15 days after culture. TNF- alpha 50 ng/ml was added into the culture of DCs from patients with hepatitis Beta and DCs were collected 48 hours after. FACS was used to detect the phenotype. The changes of cell phenotype on different days with or without TNF-alpha were tested by immunolabelling and flow cytometry analysis. IL-12 ELISA kit was applied to investigate IL-12 secretion of DCs. To perform mixed leucocyte reaction (MLR) test, DCs were cocultured with allogeneic T cells at different stimulatory ratio. RESULTS: The expression of CD83 was higher in the group with TNF-alpha than in the group without TNF-alpha. The positive rates of CD80 and CD54 were not different between the two groups. DCs from patients and normal donors with typical morphology were harvested successfully in vitro. The IL-12 level was 103.93 +/- 12.59 pg/ml in the group with TNF-alpha, higher than that in the group without TNF-alpha (11.45 +/- 2.58 pg/ml, P < 0.05). MLR test showed that when the stimulatory ratio of allogeneic T cells was 1 : 20, 1 : 50, and 1 : 100 respectively, the results were 40 835 +/- 3 480, 37 525 +/- 6 398, and 31 814 +/- 5 521 respectively in group of patient; and 49 097 +/- 2 273, 43 049 +/- 2 833, and 39 091 +/- 6 469 respectively in the group of healthy blood donors. CONCLUSION: 1.DCs of patients and of healthy persons can be cultured success fully with AIM-V when induced by GM-CSF, IL-4 and TNF-alpha in vitro. 2. Functions of DCs from patients are lower than those from healthy donors. However, they are not different in amount and morphology.