Immunosuppressive microvesicles-mimetic derived from tolerant dendritic cells to target T-lymphocytes for inflammation diseases therapy.

The utilization of extracellular vesicles (EV) in immunotherapy, aiming at suppressing peripheral immune cells responsible for inflammation, has demonstrated significant efficacy in treating various inflammatory diseases. However, the clinical application of EV has faced challenges due to their inadequate targeting ability. In addition, most of the circulating EV would be cleared by the liver, resulting in a short biological half-life after systemic administration. Inspired by the natural microvesicles (MV, as a subset of large size EV) are originated and shed from the plasma membrane, we developed the immunosuppressive MV-mimetic (MVM) from endotoxin tolerant dendritic cells (DC) by a straightforward and effective extrusion approach, in which DC surface proteins were inherited for providing the homing ability to the spleen, while αCD3 antibodies were conjugated to the MVM membranes for specific targeting of T cells. The engineered MVM carried a large number of bioactive cargos from the parental cells, which exhibited a remarkable ability to promote the induction of regulatory T cells (Treg) and polarization of anti-inflammatory M2 macrophages. Mechanistically, the elevated Treg level by MVM was mediated due to the upregulation of miR-155-3p. Furthermore, it was observed that systemic and local immunosuppression was induced by MVM in models of sepsis and rheumatoid arthritis through the improvement of Treg and M2 macrophages. These findings reveal a promising cell-free strategy for managing inflammatory responses to infections or tissue injury, thereby maintaining immune homeostasis.

Endotoxin tolerance ameliorates lipopolysaccharide/D-galactosamine-induced acute liver failure by negative regulation of the NF-κB/NLRP3 and activation of Nrf2/HO-1 via Sitr1.

Acute liver failure (ALF) is a potentially fatal disorder characterized by extensive hepatocyte necrosis and rapid decline in liver function. Numerous factors, including oxidative stress, cell death, and inflammatory responses, are associated with its pathogenesis. Endotoxin tolerance (ET) refers to the phenomenon in which the body or cells exhibit low or no response to high-dose lipopolysaccharide (LPS) stimulation after pre-stimulation with low-dose LPS. However, the specific mechanism through which ET regulates LPS/D-galactosamine (D-GalN)-induced ALF remains unclear. An ALF mouse model was established by intraperitoneal injection of D-GalN (400 mg/kg) and LPS (10 mg/kg). A low dose of LPS (0.1 mg/kg/d) was continuously administered to mice for 5 d before modeling to assess the protective effect of ET. The data from this study showed that ET alleviated the inflammatory response in mice with LPS/D-GalN-induced ALF. ET inhibited LPS-induced oxidative damage and pyroptosis in macrophages in vitro. RNA sequencing analysis showed that the NF-κB/NLRP3 pathway was linked to the anti-inflammatory and antioxidative effects of ET. Furthermore, using western blot, RT-qPCR, and immunofluorescence, we verified that ET inhibited the NF-κB/NLRP3 pathway and triggered the Nrf2/HO-1 signaling pathway to attenuate oxidative stress and cell pyroptosis. Sirt1 knockdown reversed this protective effect. In summary, our research elucidates that ET prevents ALF advancement by upregulating Sirt1 levels, triggering the Nrf2/HO-1 signaling axis, and suppressing the NF-κB/NLRP3 signaling cascade to inhibit oxidative stress and cell pyroptosis. Our results provide a mechanistic explanation for the protective effect of ET against ALF.

High normal alanine aminotransferase is an indicator for better response to antiviral therapy in chronic hepatitis B.

Background: Evidence shows people living with CHB even with a normal ALT (40U/L as threshold) suffer histological disease and there is still little research to evaluate the potential benefit of antiviral benefits in them. Methods: We retrospectively examined 1352 patients who underwent liver biopsy from 2017 to 2021 and then obtained their 1-year follow-up data to analyze. Results: ALT levels were categorized into high and low, with thresholds set at >29 for males and >15 for females through Youden's Index. The high normal ALT group showed significant histological disease at baseline (56.43% vs 43.82%, p< 0.001), and better HBV DNA clearance from treatment using PSM (p=0.005). Similar results were obtained using 2016 AASLD high normals (male >30, female >19). Further multivariate logistic analysis showed that high normal ALT (both criterias) was an independent predictor of treatment (OR 1.993, 95% CI 1.115-3.560, p=0.020; OR 2.000, 95% CI 1.055-3.793, p=0.034) Both of the models had higher AUC compared with current scoring system, and there was no obvious difference between the two models (AUC:0.8840 vs 0.8835). Conclusion: Male >30 or female >19 and Male >29 or female>15 are suggested to be better thresholds for normal ALT. Having a high normal ALT in CHB provides a potential benefit in antiviral therapy.

Human papillomavirus vaccination and contributing factors of vaccination intention among adolescents and young adults in China from a socio-ecological perspective: A cross-sectional study.

OBJECTIVES: Adolescents and young adults are the main target population for human papillomavirus (HPV). The study aimed to investigate school students' HPV vaccination intentions and explore the contributing factors from a socio-ecological perspective. DESIGN: A questionnaire survey was conducted in three secondary schools and three colleges in China. SAMPLE: A total of 1756 students aged 14-22 years participated in this study. Among the 1756 participants, 182 students have received the HPV vaccine. For the remaining 1574 students, we analyzed their HPV vaccination intentions and the influencing factors. MEASUREMENTS: Survey items for sociodemographics, knowledge and awareness of HPV, sexual intercourse and sexual knowledge, subjective socioeconomic status, self-efficacy, eHealth literacy, perceived social support from family, and the availability of HPV vaccine information were measured. RESULTS: Only 182 (10.4%) had received the HPV vaccine among the 1756 participants. Among the remaining 1574 students, the majority of the students (1403, 89.1%) were willing to receive the HPV vaccine. Binary logistic regression analysis showed that students who were female, had lower self-efficacy, scored higher on sexual knowledge, believed vaccination preventing related diseases, worried about side effects after vaccination, thought oneself at risk of contracting HPV, had higher family support, knew the availability of the HPV vaccine in Mainland China from healthcare institutions, and with family residence in rural areas were more willing to receive the HPV vaccine. CONCLUSIONS: Students had high HPV vaccination intentions while had low vaccination rate. Intrapersonal, interpersonal and institutional or community factors predicted HPV vaccination intention. Public health nurses in communities and schools could target the modifiable factors to promote students' HPV vaccine uptake.

Serum ammonia variation predicts mortality in patients with hepatitis B virus-related acute-on-chronic liver failure.

Background: Hyperammonemia is critical to the development of hepatic encephalopathy (HE) and is associated with mortality in end-stage liver disease. This study investigated the clinical value of ammonia variation in hepatitis B virus-related acute-on-chronic liver failure (HBV-ACLF) patients. Methods: A total of 276 patients with HBV-ACLF were retrospectively recruited. Patients' ammonia levels were serially documented. Baseline ammonia, Peak ammonia (highest level), and Trough ammonia (lowest level) were particularly corrected to the upper limit of normal (AMM-ULN). The primary endpoint was 28-day mortality. Results: The 28-day, 3-month, and 12-month mortality rates were 19.2, 25.7, and 28.2%, respectively. A total of 51 (18.4%) patients had overt HE (grade 2/3/4). Peak AMM-ULN was significantly higher in patients with overt HE and non-survivors compared with their counterparts (P < 0.001). Following adjustment for significant confounders, high Peak AMM-ULN was an independent predictor of overt HE (hazard ratio, 1.031, P < 0.001) and 28-day mortality (hazard ratio, 1.026, P < 0.001). The cut-off of Peak AMM-ULN was 1.8, determined by using the X-tile. Patients with Peak AMM-ULN appearing on days 1-3 after admission had a higher proportion of overt HE and mortality compared to other groups. Patients with decreased ammonia levels within 7 days had better clinical outcomes than those with increased ammonia. Conclusion: Serum Peak ammonia was independently associated with overt HE and mortality in HBV-ACLF patients. Serial serum ammonia may have prognostic value.

Association between body roundness index and non-alcoholic fatty liver disease detected by Fibroscan in America.

BACKGROUND: The prevalence of non-alcoholic fatty liver disease (NAFLD) and obesity is worldwide on the rise. Body roundness index (BRI), as a newly developed anthropometric indicator, has been recently reported to identify obesity. However, it is still unclear whether BRI is associated with the prevalence of NAFLD. METHODS: Data were from the National Health and Nutrition Examination Survey (NHANES) 2017-2018. NAFLD was diagnosed based on hepatic steatosis defined by CAP values ≥274 dB/m. Multivariable logistic regression analysis was performed to detect the association between BRI and the odds of NAFLD. Subgroup analysis stratified by age, gender, BMI, and race was further conducted. To explore the potential ability of BRI in predicting NAFLD, the area under the curve (AUC) of BRI was calculated by receiver operating characteristic (ROC) analysis. RESULTS: Among the 4467 study participants, 1718 (38.5%) were diagnosed with NAFLD. Compared to the non-NAFLD group, participants with NAFLD had a higher level of BRI. The positive associations between BRI and NAFLD were detected in all three models (mode 1: OR = 1.71, 95% CI: 1.65-1.78, p < 0.0001; mode 2: OR = 1.78, 95% CI: 1.71-1.86, p < 0.0001; mode3: OR = 1.23, 95% CI: 1.11-1.35, p < 0.0001). The positive association steadily existed in different subgroups after stratified by age, gender, and BMI. Moreover, the non-linear association between BRI and NAFLD was detected, presenting inverted U-shaped curves. Furthermore, BRI had a high predictive value (AUC = 0.807) in identifying NAFLD. CONCLUSIONS: BRI was positively associated with the prevalence of NAFLD among individuals in America, regardless of age, gender, and BMI. Besides, BRI presented a high ability for identifying NAFLD.

Evaluation of Neisseria gonorrhoeae Isolates Susceptibility to Antibiotics in Zhejiang Province Since 2007.

Objective: This study aimed to assess the drug susceptibility of clinical isolates of Neisseria gonorrhoeae to spectinomycin, ceftriaxone and azithromycin. Moreover, the temporal trends in the minimum inhibitory concentration (MIC) of five antibiotics from Zhejiang, China, are also in the scope of this study. Methods: A total of 1710 gonococcal clinical strains were collected between 2007 and 2021 from health-care institutions in Zhejiang. The MICs of ceftriaxone, azithromycin, spectinomycin, penicillin and ciprofloxacin were assessed by agar dilution method on 1710 Neisseria gonorrhoeae isolates. Count data were expressed as strains and rates, and MICs distribution was elucidated using descriptive statistics. Results: The total resistance rates of gonococci to azithromycin, spectinomycin, penicillin and ciprofloxacin in this study were 19.3%, 0.3%, 75.4% and 99.7%, respectively. Conclusion: The in vitro results showed a high prevalence of resistance to ciprofloxacin and penicillin. Azithromycin resistance rate has exceeded 5%, suggested a high prevalence of resistance. Ceftriaxone and spectinomycin are suggested based on this study for the treatment of Neisseria gonorrhoeae in Zhejiang.

Analysis of factors influencing the use of child restraint system by parents of children aged 0-6 years: an information, motivation, behavioral skills model-based cross-sectional study.

BACKGROUND: Children's injuries from traffic accidents have been identified as a global public health issue. Child restraint system (CRS) is a useful tool for lowering the risk of injury to children. Nevertheless, CRS usage is really low in China. The goal of the current study was to investigate the use of CRS after the legislation revised in China and to explore the influencing factors based on Information, Motivation, and Behavioral Skills model (IMB). METHODS: The study is a cross-sectional survey of parents who took their 0 to 6-year-old children for seeking primary care services at the Children Preventive Health Care Clinic of a tertiary hospital in Shandong Province, China. Parents were invited to complete the self-administered questionnaire between March and June 2022, including their knowledge, motivation, and behavioral skills, use behavior of CRS and socio-demographics. Ordinal logistic regression was used to explore the factors associated with CRS use by using SPSS software (version 26.0). RESULTS: In total, 442 parents participated in the study; 56.1% (n = 201) of the parents utilized CRS for their child passengers, however only 29.0% used CRS frequently. The result of logistic regression analysis show that parents with junior college (OR = 0.398, 95%CI: 0.185 ~ 0.857), possessing a high family economic status(OR = 0.225, 95%CI: 0.088 ~ 0.578), being trained on children's unintentional injuries(OR = 0.435,95%CI: 0.272 ~ 0.695), and having high scores on CRS riding mode cognition(OR = 0.476, 95%CI: 0.368 ~ 0.616), CRS type cognition(OR = 0.519, 95%CI: 0.392 ~ 0.689), CRS use motivation(OR = 0.392, 95%CI: 0.295 ~ 0.520) and installation skills(OR = 0.559, 95%CI:0.411 ~ 0.761) were the main factors promoting the usage of CRS. CONCLUSIONS: This study found that the use of CRS can be increased by improving parents' knowledge, motivation and behavior skills and hence related educational programs is necessary for increasing CRS use in China.

Regulatory Effect of JAK2/STAT3 on the Immune Function of Endotoxin-tolerant Dendritic Cells and its Involvement in Acute Liver Failure.

Background and Aims: Acute liver failure (ALF) is a potentially fatal clinical syndrome with no effective treatment. This study aimed to explore the role of Janus kinase 2/signal transducer and activator of transcription 3 (JAK2/STAT3) pathway in modulating the phenotype and immune function of endotoxin-tolerant dendritic cells (ETDCs). In addition, we explored the use of EDTCs in an experimental model of ALF and investigated the associated mechanisms. Methods: In the in vitro experiment, ETDCs were transfected with adenovirus to induce SOCS1+/+ETDCs and SOCS1-/-ETDCs. Thereafter, costimulatory molecules and mixed lymphocyte reaction were assessed. Experimental mice were randomly divided into normal control, ALF, ALF+mock-ETDCs, ALF+SOCS1+/+ETDCs, ALF+AG490, and ALF+AG490+SOCS1+/+ETDCs groups. We examined the therapeutic effect of adoptive cellular immunotherapy by tail-vein injection of target ETDCs 12 h before ALF modeling. AG490, a JAK2/STAT3 inhibitor, was used in the in vivo experiment to further explore the protective mechanism of SOCS1+/+ETDCs. Results: Compared with control ETDCs, SOCS1+/+ETDCs had lower expression of costimulatory molecules, weaker allostimulatory ability, lower levels of IL-6 and TNF-α expression and higher IL-10 secretion. SOCS1-/-ETDCs showed the opposite results. In the in vivo experiments, the ALF+SOCS1+/+ETDCs and ALF+AG490+SOCS1+/+ETDCs groups showed less pathological damage and suppressed activation of JAK2/STAT3 pathway. The changes were more pronounced in the ALF+AG490+SOCS1+/+ETDCs group. Infusion of SOCS1+/+ETDCs had a protective effect against ALF possibly via inhibition of JAK2 and STAT3 phosphorylation. Conclusions: The SOCS1 gene had an important role in induction of endotoxin tolerance. SOCS1+/+ETDCs alleviated lipopolysaccharide/D-galactosamine-induced ALF by downregulating the JAK2/STAT3 signaling pathway.

The association between non-alcoholic fatty liver disease and serum ferritin levels in American adults.

BACKGROUND: Elevated serum ferritin levels (SFLs) was previously reported to be related with hepatic histologic severity and advanced liver fibrosis among non-alcoholic fatty liver disease (NAFLD) patients. However, whether NAFLD influences SFLs remains uncertain and needs more clinical evidences. This study explored the differences of SFLs in US adults with or without NAFLD. METHODS: We conducted a cross-sectional study of 3689 participants aged 18-80 years using the National Health and Nutrition Examination Survey (NHANES) 2017-2018 cycle. NAFLD status was confirmed based on controlled attenuation parameter (CAP) values ≥274 dB/m through vibration controlled and transient elastography (VCTE). We performed weighted multivariable logistic regression models to evaluate the associations between NAFLD and SFLs in different age and gender. RESULTS: There was a positive association between NAFLD and SFLs in all three models (model 1:ß = 23.07, 95% CI: 10.32, 35.81; model 2:ß = 23.68, 95% CI: 10.86, 36.50; model 3:ß = 13.86, 95% CI: 0.29, 27.43). After adjusting for the covariates, this positive association persisted in females (ß = 16.22, 95% CI: 2.81, 29.62). Further, relationships between NAFLD and SFLs were significantly different in various age groups. In the subgroup stratified by gender, their associations further differed. In males, the positive association was more prominent in 50-64 age group (ß = 70.89, 95% CI: 25.14, 116.64). In females, this positive association was more prominent in 18-34 age group (ß = 20.72, 95% CI: 7.45, 33.99). However, no correlations between severe steatosis, significant fibrosis, advanced fibrosis, cirrhosis, and SFLs in adults with NAFLD were found. CONCLUSION: This study indicated that US adults suffered with NAFLD had significantly higher SFLs compared with their counterparts in non-NAFLD group. Moreover, the associations between NAFLD and SFLs further differed by age and gender.

Tailoring the physicochemical properties of nanomaterials for immunomodulation.

Immunomodulation is poised to revolutionize the treatment of cancer, autoimmune diseases, and many other inflammation-related disorders. The immune system in these conditions can be either activated or suppressed by nanocarriers loaded with bioactive molecules. Although immunomodulation via these therapeutics has long been recognized, and a broad range of nanocarriers have been designed to accommodate varied usages, less studies have focused on the effects of nanomaterial physicochemical properties on immune responses, especially the immunity altered by nanocarrier materials alone. Conclusions are sometimes seemly inconsistent due to the complexities of nanomaterials and the immune system. An in-depth understanding of the nanocarrier-induced immune responses is essential for clinical applications. In this review, we summarize recent studies of the immune responses influenced by nanomaterial physicochemical properties with an emphasis on the intrinsic features of nanomaterials that modulate the innate and adaptive immunities. We then provide our perspectives on the design of nanomaterials for immunomodulation.

Assessment of the effectiveness of parent-targeted interventions for the use of child restraint systems: a systematic review and meta-analysis.

Background: It is essential to implement parent-targeted interventions to increase the use of child restraint systems (CRS) and thus reduce the injuries and deaths of children due to motor vehicle collisions. To optimize future intervention designs, this meta-analysis sought to quantify the effects of parent-targeted interventions and explore potential intervention moderators. Methods: Studies met inclusion criteria if they included a parents-targeted intervention that focused on increasing CRS use for children, published from the inception of the databases to January 2022, were systematically retrieved from the PubMed, Embase, Cochrane library, Web of Science, Sinomed, Wanfang, and CNKI databases. Next, 2 researchers independently screened the retrieved articles, evaluated their quality according to the Cochrane Tool, and extracted the data. Finally, Stata12.0 was used for the meta-analysis. Heterogeneity was examined with I2, stratified analyses, and meta-regression. Results: Of the 1,690 articles retrieved, 9 studies, comprising 22,329 parents of children aged 0-12 years, were ultimately included in the analysis. The results of the meta-analysis showed that the CRS use rate of the intervention group was 1.62 times higher than that of the control group [95% confidence interval (CI): 1.25-2.11, Z=3.616, P<0.001], indicating the positive effect of parent-targeted interventions on promoting the use of CRS. The subgroup analysis found that interventions guided by behavioral theories increased the use of CRS (odds ratio: 1.44, 95% CI: 1.27-1.63, n=5). The difference in the use of CRS between the groups in the studies that were not guided by theories was not statistically significant, indicating that interventions guided by behavioral theories may be the source of the heterogeneity. Risk of bias was low in most studies. Conclusions: It is necessary to conduct interventions with parents to increase the use of CRS. The effects on CRS use appear to differ depending on whether the interventions are guided by behavioral theories. In-depth research needs to be conducted to explore the characteristics of the interventions, especially those guided by different behavioral theories, to reduce child vehicle injuries.

Single cell sequencing analysis identifies genetics-modulated ORMDL3+ cholangiocytes having higher metabolic effects on primary biliary cholangitis.

BACKGROUND: Primary biliary cholangitis (PBC) is a classical autoimmune disease, which is highly influenced by genetic determinants. Many genome-wide association studies (GWAS) have reported that numerous genetic loci were significantly associated with PBC susceptibility. However, the effects of genetic determinants on liver cells and its immune microenvironment for PBC remain unclear. RESULTS: We constructed a powerful computational framework to integrate GWAS summary statistics with scRNA-seq data to uncover genetics-modulated liver cell subpopulations for PBC. Based on our multi-omics integrative analysis, 29 risk genes including ORMDL3, GSNK2B, and DDAH2 were significantly associated with PBC susceptibility. By combining GWAS summary statistics with scRNA-seq data, we found that cholangiocytes exhibited a notable enrichment by PBC-related genetic association signals (Permuted P < 0.05). The risk gene of ORMDL3 showed the highest expression proportion in cholangiocytes than other liver cells (22.38%). The ORMDL3+ cholangiocytes have prominently higher metabolism activity score than ORMDL3- cholangiocytes (P = 1.38 × 10-15). Compared with ORMDL3- cholangiocytes, there were 77 significantly differentially expressed genes among ORMDL3+ cholangiocytes (FDR < 0.05), and these significant genes were associated with autoimmune diseases-related functional terms or pathways. The ORMDL3+ cholangiocytes exhibited relatively high communications with macrophage and monocyte. Compared with ORMDL3- cholangiocytes, the VEGF signaling pathway is specific for ORMDL3+ cholangiocytes to interact with other cell populations. CONCLUSIONS: To the best of our knowledge, this is the first study to integrate genetic information with single cell sequencing data for parsing genetics-influenced liver cells for PBC risk. We identified that ORMDL3+ cholangiocytes with higher metabolism activity play important immune-modulatory roles in the etiology of PBC.

Changes and Clinical Significance of PIVKA-II in Hepatitis E Patients.

Increased protein induced by vitamin K absence or antagonist-II (PIVKA-II) levels had been widely reported in patients with hepatocellular carcinoma (HCC) and chronic hepatitis. However, the role of PIVKA-II in hepatitis E is unclear. The aim of this study was to clarify the changes related with PIVKA-II and its clinical significance in hepatitis E. We enrolled 84 patients with hepatitis E hospitalized in two hospitals from December 2019 to June 2021. The levels of serum PIVKA-II and related serological indicators in the patients were determined to elucidate the role of PIVKA-II in hepatitis E. We observed that 59.51% (50/84) of patients showed an increase in PIVKA-II levels. Compared with the normal PIVKA-II group (<32 mAU/L), patients in the elevated PIVKA-II group (>32 mAU/L) had much higher serum total bilirubin (TBIL), direct bilirubin (DBIL), indirect bilirubin (IBIL), and total bile acid (TBA) levels (p < 0.05 for each). Compared with the slightly elevated PIVKA-II group (32-125 mAU/L), patients in the significantly elevated PIVKA-II group (>125 mAU/L) had much lower serum albumin, alanine aminotransferase (ALT), aspartate transaminase (AST) levels, and longer days for the hospital stay (p < 0.05 for each). The association of PIVKA-II with TBIL and DBIL was an inverted U-shaped curve with an inflection point at 199.1 mAU/L). The association of PIVKA-II with IBIL was a U-shaped curve with an inflection point at 18.6 mAU/L while the association of PIVKA-II with albumin was an inverted U-shaped curve with an inflection point at 18.6 mAU/L. With the improvement of the disease, PIVKA-II levels were gradually decreased and finally returned to normal. This trend was consistent with that of bilirubin, and a peak appeared in the third week. Therefore, findings from our study show that the increase in PIVKA-II levels can be related to the degree of hepatic insufficiency in patients with hepatitis E, wherein PIVKA-II levels are transiently increased, and the trend of change can be related to the disease course.

In-flight transmission cluster of COVID-19: a retrospective case series.

BACKGROUND: No data is available about in-flight transmission of SARS-CoV-2. Here, we report an in-flight transmission cluster of COVID-19 and describe the clinical characteristics of these patients. METHODS: After a flight, laboratory-confirmed COVID-19 was reported in 12 patients. Ten patients were admitted to the designated hospital. Data was collected from 25th January to 28th February 2020. Clinical information was retrospectively collected. RESULTS: All patients were passengers, and none were flight attendants. The median age was 33 years, and 70% were females. None was admitted to intensive care unit, and no patients died up to 28th February. The median incubation period was 3.0 days and time from onset of illness to hospital admission was 2 days. The most common symptom was fever. Two patients were asymptomatic and had normal chest CT scan during hospital stay. On admission, initial RT-PCR was positive in 9 patients, and initial chest CT was positive in half of the patients. The median lung 'total severity score' of chest CT was 6. 'Crazy-paving' pattern, pleural effusion, and ground-glass nodules were seen. CONCLUSION: There is potential for COVID-19 transmission in aeroplanes, but the symptoms were mild in our patients. Passengers and attendants must be protected during flights.

Interleukin 10 Gene-Modified Bone Marrow-Derived Dendritic Cells Attenuate Liver Fibrosis in Mice by Inducing Regulatory T Cells and Inhibiting the TGF-ß/Smad Signaling Pathway.

AIM: To explore the therapeutic effects and mechanisms of interleukin 10 gene-modified bone marrow-derived dendritic cells (DC-IL10) on liver fibrosis. METHODS: In vitro, BMDCs were transfected with lentiviral-interleukin 10-GFP (LV-IL10-GFP) at the MOI of 1 : 40. Then, the phenotype (MHCII, CD80, and CD86) and allo-stimulatory ability of DC-IL10 were identified by flow cytometry, and the levels of IL-10 and IL-12 (p70) secreted into the culture supernatants were quantified by ELISA. In vivo, DC-IL10 was injected into mice with CCl4-induced liver fibrosis through the tail vein. Lymphocytes were isolated to investigate the differentiation of T cells, and serum and liver tissue were collected for biochemical, cytokine, histopathologic, immune-histochemical, and Western blot analyzes. RESULTS: In vitro, the expressions of MHCII, CD80, and CD86 in DC-IL10 were significantly suppressed, allogeneic CD4+T cells incubated with DC-IL10 showed a lower proliferative response, and the levels of IL-10 and IL-12 (p70) secreted into the DC-IL10 culture supernatants were significantly increased and decreased, respectively. In vivo, regulatory T cells (Tregs) were significantly increased, while ALT, AST, and inflammatory cytokines were significantly reduced in the DC-IL10 treatment group, and the degree of hepatic fibrosis was obviously reversed. The TGF-ß/smad pathway was inhibited following DC-IL10 treatment compared to the liver fibrosis group. CONCLUSION: IL-10 genetic modification of BMDCs may maintain DC in the state of tolerance and allow DC to induce T cell hyporesponsiveness or tolerance. DC-IL10 suppressed liver fibrosis by inducing Treg production and inhibiting the TGF-ß/smad signaling pathway.

Dendritic cells with increased expression of suppressor of cytokine signaling 1(SOCS1) gene ameliorate lipopolysaccharide/d-galactosamine-induced acute liver failure.

Acute liver failure is a devastating clinical syndrome with extremely terrible inflammation reaction, which is still lack of effective treatment in clinic. Suppressor of Cytokine Signaling 1 protein is inducible intracellular negative regulator of Janus kinases (JAK)/signal transducers and activators of transcription (STAT) pathway that plays essential role in inhibiting excessive intracellular signaling cascade and preventing autoimmune reaction. In this paper, we want to explore whether dendritic cells (DCs) with overexpression of SOCS1 have a therapeutic effect on experimental acute liver failure. Bone marrow derived dendritic cells were transfected with lentivirus encoding SOCS1 and negative control lentivirus, thereafter collected for costimulatory molecules analysis, allogeneic Mixed Lymphocyte Reaction and Western blot test of JAK/STAT pathway. C57BL/6 mice were randomly separated into normal control and treatment groups which respectively received tail vein injection of modified DCs, negative control DCs and normal saline 12 h earlier than acute liver failure induction. Our results indicated that DCs with overexpression of SOCS1 exhibited like regulatory DCs (DCregs) with low level of costimulatory molecules and poor allostimulatory ability in vitro, which was supposed to correlate with block of JAK2/STAT1 signaling. In vivo tests, we found that infusion of modified DCs increased survival rate of acute liver failure mice and alleviate liver injury via inhibition of TLR4/HMGB1 pathway. We concluded that DCs transduced with SOCS1 gene exhibit as DCregs through negative regulation of JAK2/STAT1 pathway and ameliorated lipopolysaccharide/d-galactosamine induced acute liver failure via inhibition of TLR4 pathway.

Aberrant KIF20A expression might independently predict poor overall survival and recurrence-free survival of hepatocellular carcinoma.

Kinesin family member 20A (KIF20A) is an essential regulator of cytokinesis. In this study, by performing a retrospective study based on data from the Cancer Genome Atlas (TCGA)-Liver and Hepatocellular Carcinoma (LIHC) cohort, we tried to assess the independent prognostic value of KIF20A in terms of overall survival (OS) and recurrence-free survival (RFS). Results showed that normal liver tissues had very low KIF20A expression compared with normal tissues in other cohorts in TCGA. However, the primary HCC tissues (N = 371) had significantly elevated KIF20A expression than normal liver tissues (N = 50). Immunohistochemistry (IHC) data showed that normal hepatocytes had weak KIF20A staining. In comparison, some HCC tissues had medium and strong KIF20A expression, with nuclear-enhanced staining. By grouping patients with primary HCC (N = 365) into high and low KIF20A expression groups, we found that the high expression group had a substantially higher proportion of high-grade tumors (G3/G4) (34/65, 52.3% vs. 96/295, 32.5%, P = 0.0027), advanced tumors (stage III/IV) (28/61, 45.9% vs. 59/280, 21.1%, P < 0.0001) and death (44/67, 65.7% vs. 86/298, 28.9%, P < 0.0001) compared with the low expression group. Kaplan-Meier curves of OS and RFS indicated that high KIF20A expression was associated with worse survival outcomes. Subgroup analysis confirmed the associations in G1/G2, G3/G4 tumors and in early and advanced stages. Following univariate and multivariate analysis revealed that KIF20A expression was an independent prognostic indicator for poor OS (HR: 1.304, 95%CI: 1.157-1.469, P < 0.001) and RFS (HR: 1.144, 95%CI: 1.028-1.272, P < 0.001). Based on these findings, we infer that KIF20A was aberrantly expressed in HCC tissues and its expression might independently predict poor OS and RFS. © 2018 IUBMB Life, 70(4):328-335, 2018.

Deregulation of Regulatory T Cells in Acute-on-Chronic Liver Failure: A Rat Model.

Aims. Acute-on-chronic liver failure (ACLF) and acute liver failure (ALF) are similar in many respects during their acute exacerbation; however, ACLF generally has a poorer prognosis. We aimed to investigate the role and dynamic changes of regulatory T cell (Treg) and T helper 17 (Th17) cell proportions during ACLF progress. Methods. All rats were classified into two groups randomly: ACLF group and ALF group (control group). The rat model of ACLF was preestablished by intraperitoneal injection of carbon tetrachloride for 2 months. Then acute liver injury was induced by combined D-galactosamine and lipopolysaccharide. Six time points were examined before or after acute induction. Liver samples were performed with hematoxylin-eosin and Masson staining; circulatory Treg and Th17 cell frequencies were determined using flow cytometry assays; serum levels of alanine aminotransferase, aspartate aminotransferase, interleukin-10 (IL-10), and interferon-γ (IFN-γ) were examined. Results. In group ACLF, both Th17 cell proportion and IFN-γ level presented upgrade firstly and then descend latter tendency; the trends of Treg cell proportion and IL-10 level were observed to gradually decrease and became stable. Conclusion. The Treg cells played an important role in the immunologic mechanism during the process of ACLF. And the function of Treg cells in ACLF was defective.