A recombinant IL-1ß vaccine attenuates bleomycin-induced pulmonary fibrosis in mice.

Interleukin-1ß (IL-1ß) contributes to interstitial lung disease (ILD) and pulmonary fibrosis (PF), thus representing a potential therapeutic target for PF. In this study, we first verified the increased expression of IL-1ß in human fibrotic lung specimens and mouse lung tissues after intratracheal (i.t.) instillation of bleomycin (BLM), after which the pro-inflammatory and pro-fibrotic effects of recombinant IL-1ß were tested in mice. The results above suggested that vaccination against IL-1ß could be an effective strategy for managing PF. An anti-IL-1ß vaccine (PfTrx-IL-1ß) was designed by incorporating two IL-1ß-derived polypeptides, which have been verified as the key domains that mediate the binding of IL-1ß to its type I receptor, into Pyrococcus furiosus thioredoxin (PfTrx). The fusion protein PfTrx-IL-1ß was prepared by using E. coli expression system. The vaccine was well tolerated; it induced robust and long-lasting antibody responses in mice and neutralized the biological activity of IL-1ß, as shown in cellular assays. Pre-immunization with PfTrx-IL-1ß effectively protected mice from BLM-induced lung injury, inflammation, and fibrosis. In vitro experiments further showed that anti-PfTrx-IL-1ß antibodies counteracted the effects of IL-1ß concerning pro-inflammatory and pro-fibrotic cytokine production by primary mouse lung fibroblast, macrophages (RAW264.7), and type II alveolar epithelial cell (A549), primary mouse lung fibroblast activation and epithelial-mesenchymal transition (EMT) of alveolar epithelial cells. In addition, the vaccination did not compromise the anti-infection immunity in mice, as validated by a sepsis model. Our preliminary study suggests that the anti-IL-1ß vaccine we prepared has the potential to be developed as a therapeutic measure for PF. Further experiments are warranted to evaluate whether IL-1ß vaccination has the capacity of inhibiting chronic progressive PF and reversing established PF.

The relationship between perfectionism, self-perception of orofacial appearance, and mental health in college students.

Background: In dental clinical practice, self-perception of orofacial appearance is highly correlated with treatment satisfaction. Therefore, it is important to explore factors correlated with self-perception of orofacial appearance. Perfectionism may be one such factor. This study investigated the role of perfectionism in self-perception of orofacial appearance. Methods: Participants completed an online questionnaire that included demographic data, a measure of perfectionism, a measure of self-perception of orofacial appearance (including body image, smile appearance concern, and self-esteem), and a measure of anxiety and depression. Results: High perfectionism scores significantly predicted greater age, body image, smile appearance concern, and mental health scores and lower self-esteem scores (p < 0.005). After adjusting for possible confounding variables, smile appearance concern largely disappeared. Mental health acted as a mediator in the relationships between perfectionism and three orofacial appearance characteristics. Conclusion: High perfectionism predicted higher self-perception of body image, and lower mental health and self-esteem in college students. Mental health could mediate the relationships between perfectionism and self-perception of orofacial appearance.

The rapid inhibition of B-cell activation markers by belimumab was associated with disease control in systemic lupus erythematosus patients.

Objective: To examine the kinetics of B cell subsets and activation markers in the early stage of belimumab treatment and their correction with treatment response. Methods: We enrolled 27 systemic lupus erythematosus (SLE) patients receiving 6 months belimumab treatment. Flow cytometry was used to test their B cell subsets and activation markers (including CD40, CD80, CD95, CD21low, CD22, p-SYK and p-AKT). Results: During belimumab treatment, SLEDAI-2K declined, the proportions of CD19+ B cells and naïve B cells decreased, whereas the switched memory B cells and non-switched B cells increased. The larger variations of the B cell subsets and the activation markers were in the first 1 month than the other later time frames. The ratio of p-SYK/p-AKT on non-switched B cell at 1 month was associated with the SLEDAI-2K decline rate in the 6 months of belimumab treatment. Conclusion: B cell hyperactivity was rapidly inhibited in the early stage of belimumab treatment, and the ratio of p-SYK/p-AKT may predict SLEDAI-2K decline. Clinical Trial Registration: https://www.clinicaltrials.gov/ct2/show/NCT04893161?term=NCT04893161&draw=2&rank=1; identifier: NCT04893161.

A new genus and species of leptonetid spiders (Araneae, Leptonetidae) from Guangdong Province, China.

BACKGROUND: The spider family Leptonetidae Simon, 1890 includes 20 genera and 366 species from North America, the Mediterranean Region and Asia. Currently, 132 species belonging to six genera have been recorded in China. NEW INFORMATION: A new genus and species of leptonetid spiders, Yueleptonetadongxing gen. et sp. n., is described from Guangdong Province, China. Yueleptoneta gen. n. is distinct from the other genera in the chelicerae having the stridulatory file on the lateral margin and the male palp having a tarsal spur, lacking strong spines or apophyses on the femur and tibia.

The association between problematic smartphone use and the severity of temporomandibular disorders: A cross-sectional study.

Objective: To evaluate the prevalence of different types of temporomandibular disorders (TMD) symptoms in young adults and determine their associations with problematic smartphone use (PSU). Methods: The data of the study were collected from local university students through an online questionnaire survey. Demographic information, Fonseca Anamnestic Index (FAI), Smartphone Addiction Scale-Short Version (SAS-SV), and Patient Health Questionnaire-4 (PHQ-4) responses were gathered electronically and analyzed using multiple logistic regression analysis. Results: There were 163 male and 307 female respondents were participated in this study. The prevalence of PSU and TMD were 83.6% and 66.4%, respectively. There was a moderate statistical correlation between PSU and TMD among young adults (r = 0.31, p < 0.01). The logistic regression model revealed that the risk of TMD was 1.77 times higher in people with PSU than in those without PSU (OR = 1.77; 95% CI 1.04-3.06). PSU is a risk factor for pain-related TMD (OR = 1.81; 95% CI 1.08-3.04) but not intra-articular TMD. Conclusion: Subjects showed high prevalence of both TMD and PSU. People with PSU experienced more severe and frequent pain-related rather than intra-articular TMD symptoms than those without PSU. By reducing the problematic smartphone use, the risk factor of TMD might be avoided.

HIP/PAP protects against bleomycin-induced lung injury and inflammation and subsequent fibrosis in mice.

Hepatocarcinoma-intestine-pancreas/pancreatitis-associated protein (HIP/PAP), a C-type lectin, exerts anti-oxidative, anti-inflammatory, bactericidal, anti-apoptotic, and mitogenic functions in several cell types and tissues. In this study, we explored the role of HIP/PAP in pulmonary fibrosis (PF). Expression of HIP/PAP and its murine counterpart, Reg3B, was markedly increased in fibrotic human and mouse lung tissues. Adenovirus-mediated HIP/PAP expression markedly alleviated bleomycin (BLM)-induced lung injury, inflammation, and fibrosis in mice. Adenovirus-mediated HIP/PAP expression alleviated oxidative injury and lessened the decrease in pulmonary superoxide dismutase (SOD) activity in BLM-treated mice, increased pulmonary SOD expression in normal mice, and HIP/PAP upregulated SOD expression in cultured human alveolar epithelial cells (A549) and human lung fibroblasts (HLF-1). Moreover, in vitro experiments showed that HIP/PAP suppressed the growth of HLF-1 and ameliorated the H2 O2 -induced apoptosis of human alveolar epithelial cells (A549 and HPAEpiC) and human pulmonary microvascular endothelial cells (HPMVEC). In HLF-1, A549, HPAEpiC, and HPMVEC cells, HIP/PAP did not affect the basal levels, but alleviated the TGF-ß1-induced down-regulation of the epithelial/endothelial markers E-cadherin and vE-cadherin and the over-expression of mesenchymal markers, such as α-SMA and vimentin. In conclusion, HIP/PAP was found to serve as a potent protective factor in lung injury, inflammation, and fibrosis by attenuating oxidative injury, promoting the regeneration of alveolar epithelial cells, and antagonizing the pro-fibrotic actions of the TGF-ß1/Smad signaling pathway.

Hepatocarcinoma-intestine-pancreas/pancreatitis-associated protein (HIP/PAP) confers protection against hepatic fibrosis through downregulation of transforming growth factor ß receptor II.

Hepatocarcinoma-intestine-pancreas/pancreatitis-associated protein (HIP/PAP) has antimicrobial, antioxidant, anti-inflammatory, mitogenic, and antiapoptotic effects and thus exerts important functions in the maintenance of integrity and homeostasis of several organs, such as the gastrointestinal tract, pancreas, and liver. Although the potent hepatoprotective effect of HIP/PAP has been validated, its impact on liver fibrosis has not been reported. In this study, we evaluated the role of HIP/PAP on hepatic fibrosis and explored the possible underlying mechanisms. We found that the expression of HIP/PAP and its mouse counterpart, Reg3B, was markedly upregulated in fibrotic human or mouse livers. Intraperitoneal (i.p.) interleukin (IL)-10, IL-6, and TNF-α but not TGF-ß1 significantly induced hepatic overexpression of Reg3B in mice. In both CCl4 and BDL liver fibrosis models, adenovirus-mediated ectopic expression of HIP/PAP markedly alleviated liver injury, inflammation, collagen deposition, hepatic stellate cell activation, and the overexpression of profibrotic cytokines, including transforming growth factor ß1 (TGF-ß1), platelet-derived growth factor (PDGF)-A, B, connective tissue growth factor (CTGF), and plasminogen activator inhibitor-1 (PAI-1), in mice. In vitro experiments demonstrated that, in addition to suppressing hepatic stellate cell proliferation and accelerating hepatocyte proliferation, HIP/PAP mitigated TGF-ß1-induced hepatic stellate cell activation, hepatocyte epithelial-mesenchymal transition (EMT) and upregulated expression of profibrotic cytokines in both hepatic stellate cells and hepatocytes. Moreover, HIP/PAP attenuated the overexpression of TGF-ß receptor II (TGF-ßRII) in fibrotic mouse livers and decreased the basal expression of TGF-ßRII in nonfibrotic mouse livers as well as in cultured hepatocytes and hepatic stellate cells, which is at least partly attributable to the TGF-ß1-antagonizing function of HIP/PAP. This study indicates that increased expression of hepatic HIP/PAP serves as a countermeasure against liver injury and fibrosis. Exogenous supplementation of HIP/PAP might be a promising therapeutic agent for hepatic fibrosis as well as liver injury.

Thymosin ß4 suppresses CCl4 -induced murine hepatic fibrosis by down-regulating transforming growth factor ß receptor-II.

BACKGROUND: The present study aimed to clarify the effects of thymosin ß4 (Tß4) on CCl4 -induced hepatic fibrosis in mice and to further explore the underlying mechanisms. METHODS: Expression of Tß4 in fibrotic liver tissues was assessed by a quantitative real time-reverse transcriptase polymerase chain reaction and immunohistochemistry. The effects of intraperitoneal adeno-associated virus-Tß4 (AAV-Tß4) on CCl4 -induced hepatic fibrosis were observed by the evaluation of collagen deposition, hepatic stellate cell (HSC) activation and pro-fibrotic cytokine expression. In vitro tests with HSCs and hepatocytes were performed to confirm the effects of Tß4. RESULTS: The expression of Tß4 was down-regulated in fibrotic mouse livers but was rapidly up-regulated by CCl4 -induced acute injury. AAV-Tß4 pre-treatment significantly attenuated liver injury, collagen deposition, HSC activation and pro-fibrotic cytokine over-expression, such as transforming growth factor ß1 (TGF-ß1), platelet-derived growth factor B (PDGF-B), connective tissue growth factor (CTGF) and plasminogen activator inhibitor-1 (PAI-1) in CCl4 -intoxicated mouse livers. In vitro experiments showed that Tß4 suppressed HSC proliferation, blunted TGF-ß1-induced HSC activation and reduced TGF-ß1-induced TGF-ß1, PDGF-B, CTGF and PAI-1 expression in both HSCs and hepatocytes. Ectopic Tß4 ameliorated the over-expression of TGF-ß receptor-II (TGF-ßRII) in the fibrotic mouse livers. Exogenous Tß4 down-regulated TGF-ßRII expression, whereas neutralizing endogenous extracellular Tß4 with a specific antibody up-regulated TGF-ßRII expression in cultured HSCs and hepatocytes. CONCLUSIONS: Tß4 possesses anti-fibrotic activity in the liver, which is attributable, at least partly, to down-regulating TGF-ßRII and thereby blunting TGF-ß1-mediated fibrogenetic signaling in both HSCs and hepatocytes.

[PDGF-B immunogen preparation and the suppressive effect of anti-PDGF-B ascite antibody on the proliferation of hepG2 cells].

To observe the immunogenicity of hPDGF-B immunogens that were synthesized with the fusional expression vector pET28-Trx and to test the suppressive effect of these specific antibodies induced by both of immunogens on proliferation of human HepG2 hepatoma cells. First, we chose 2 antigenic epitopes hPDGF-BΔ103-118aa and hPDGF-BΔ152-167aa from human PDGF-B and inserted these 2 coding regions into the empty vector plasmid pET28-Trx, separately. Second, mice were immunized with purified recombinant proteins to generate polyclonal antibody. Then we intraperitoneally injected mice bearing hepatoma 22 (H22) tumor cells to prepare antibody ascites. ELISA and Western blot were used to detect the titer and the utility of the antibody, respectively. Finally, HepG2 cells were exposed to PDGF-BB protein or anti-PDGF-B ascite antibody in different dilution concentrations groups and the proliferation of HepG2 cells was quantified by CCK8 assay. As the results, we identified mice that could produce high drop of neutralizing antibodies against hPDGF-B induced by both two recombinant proteins. Two anti-PDGF-B ascite antibodies could markedly inhibit the proliferation of HepG2 cells by blocking the stimulating effect of PDGF-BB protein. Our results suggest that Trx-PDGF-B recombinant protein as immunogen provides a new method for the preparation of PDGF-B vaccine, and also a new idea for the treatment of hepatocellular carcinoma in clinical practice.

A flux-gradient system for simultaneous measurement of the CH4, CO2, and H2O fluxes at a lake-air interface.

Inland lakes play important roles in water and greenhouse gas cycling in the environment. This study aims to test the performance of a flux-gradient system for simultaneous measurement of the fluxes of water vapor, CO2, and CH4 at a lake-air interface. The concentration gradients over the water surface were measured with an analyzer based on the wavelength-scanned cavity ring-down spectroscopy technology, and the eddy diffusivity was measured with a sonic anemometer. Results of a zero-gradient test indicate a flux measurement precision of 4.8 W m(-2) for water vapor, 0.010 mg m(-2) s(-1) for CO2, and 0.029 µg m(-2) s(-1) for CH4. During the 620 day measurement period, 97%, 69%, and 67% of H2O, CO2, and CH4 hourly fluxes were higher in magnitude than the measurement precision, which confirms that the flux-gradient system had adequate precision for the measurement of the lake-air exchanges. This study illustrates four strengths of the flux-gradient method: (1) the ability to simultaneously measure the flux of H2O, CO2, and CH4; (2) negligibly small density corrections; (3) the ability to resolve small CH4 gradient and flux; and (4) continuous and noninvasive operation. The annual mean CH4 flux (1.8 g CH4 m(-2) year(-1)) at this hypereutrophic lake was close to the median value for inland lakes in the world (1.6 g CH4 m(-2) year(-1)). The system has adequate precision for CH4 flux for broad applications but requires further improvement to resolve small CO2 flux in many lakes.