Targeted Gene Insertion and Replacement in the Basidiomycete Ganoderma lucidum by Inactivation of Nonhomologous End Joining Using CRISPR/Cas9.

Targeted gene insertion or replacement is a promising genome-editing tool for molecular breeding and gene engineering. Although CRISPR/Cas9 works well for gene disruption and deletion in Ganoderma lucidum, targeted gene insertion and replacement remain a serious challenge due to the low efficiency of homologous recombination (HR) in this species. In this work, we demonstrate that the DNA double-strand breaks induced by Cas9 were mainly repaired via the nonhomologous end joining (NHEJ) pathway, at a frequency of 96.7%. To establish an efficient target gene insertion and replacement tool in Ganoderma, we first inactivated the NHEJ pathway via disruption of the Ku70 gene (ku70) using a dual single guide RNA (sgRNA)-directed gene deletion method. Disruption of the ku70 gene significantly decreased NHEJ activity in G. lucidum. Moreover, ku70 disruption strains exhibited 96.3% and 93.1% frequencies of targeted gene insertion and replacement, respectively, when target DNA with the orotidine 5'-monophosphate decarboxylase (ura3) gene and 1.5-kb homologous 5'- and 3'-flanking sequences was used as a donor template, compared to 3.3% and 0%, respectively, at these targeted sites for a control strain (Cas9 strain). Our results indicated that ku70 disruption strains were efficient recipients for targeted gene insertion and replacement. This tool will advance our understanding of functional genomics in G. lucidum. IMPORTANCE Functional genomic studies in Ganoderma have been hindered by the absence of adequate genome-engineering tools. Although CRISPR/Cas9 works well for gene disruption and deletion in G. lucidum, targeted gene insertion and replacement have remained a serious challenge due to the low efficiency of HR in these species, although such precise genome modifications, including site mutations, site-specific integrations, and allele or promoter replacements, would be incredibly valuable. In this work, we inactivated the NHEJ repair mechanism in G. lucidum by disrupting the ku70 gene using the CRISPR/Cas9 system. Moreover, we established a target gene insertion and replacement method in ku70-disrupted G. lucidum that possessed high-efficiency gene targeting. This technology will advance our understanding of the functional genomics of G. lucidum.

Highly Stretchable, Tough, Resilient, and Antifatigue Hydrogels Based on Multiple Hydrogen Bonding Interactions Formed by Phenylalanine Derivatives.

Noncovalent cross-linked hydrogels with promising mechanical properties are on demand for applications in tissue engineering, flexible electronics, and actuators. However, integrating excellent mechanical properties with facile preparation for the design of hydrogen bond cross-linked hydrogels is still challenging. In this work, an advanced hydrogel was prepared from acrylamide and N-acryloyl phenylalanine by one-pot free-radical copolymerization. Owing to hydrophobicity-assisted multiple hydrogen bonding interactions among phenylalanine derivatives, the hydrogels exhibited fascinating mechanical behaviors: tensile strength of 0.35 MPa, elongation at break of 2100%, tearing energy of 1134 J/m2, and compression strength of 3.56 MPa. The hydrogels also showed robust elasticity and fatigue resistance, and the compression strength did not show any decline, even after 100 successive cycles, as well as promising self-recovery property. In addition, the cytotoxicity test in vitro proved that the hydrogel showed good biocompatibility with normal human liver cells (LO2 cells). The excellent stretchability, robust elasticity, high toughness, fatigue resistance, and biocompatibility of the hydrogel demonstrated its vast potential in the biomedical field and flexible electronic devices.

Helicobacter hepaticus infection-induced IL-33 promotes hepatic inflammation and fibrosis through ST2 signaling pathways in BALB/c mice.

It has been documented that Helicobacter hepaticus (H. hepaticus) infection is linked to hepatic inflammation and fibrosis. Interleukin 33 (IL-33) is a cytokine involved in inflammatory and fibrotic diseases, but its relevance to H. hepaticus infection-induced liver inflammation and fibrosis is unknown. In this study, we found that the expression of IL-33 in mice liver was significantly induced by H. hepaticus infection at 24 weeks post infection (WPI). Immunohistochemistry analysis revealed that IL-33 was transferred from the nucleus to the cytoplasm due to infection. The quantitation of inflammatory cytokine and histopathology evaluation showed that IL-33 knockdown attenuated the H. hepaticus-induced hepatic inflammation and fibrosis. More importantly, H. hepaticus promoted the expression of the IL-33 receptor ST2 on cell surfaces, and the expression of ST2 then activated the expression nuclear factor-κB (p65), α-SMA, and Erk1/2. These observations provide novel insights into the pathogenic mechanism of hepatic inflammation and fibrosis during H. hepaticus infection.

A Large-Scale Multicenter Study Validates Aldo-Keto Reductase Family 1 Member B10 as a Prevalent Serum Marker for Detection of Hepatocellular Carcinoma.

Aldo-keto reductase family 1 member B10 (AKR1B10) is a secretory protein overexpressed in hepatocellular carcinoma (HCC). We aimed to evaluate AKR1B10 as a serum marker for detection of HCC. Herein, we conducted a cohort study that consecutively enrolled 1,244 participants from three independent hospitals, including HCC, healthy controls (HCs), benign liver tumors (BLTs), chronic hepatitis B (CHB), and liver cirrhosis (LC). Serum AKR1B10 was tested by time-resolved fluorescent assays. Data were plotted for receiver operating characteristic (ROC) curve analyses. Alpha-fetoprotein (AFP) was analyzed for comparison. An exploratory discovery cohort demonstrated that serum AKR1B10 increased in patients with HCC (1,567.3 ± 292.6 pg/mL; n = 69) compared with HCs (85.7 ± 10.9 pg/mL; n = 66; P < 0.0001). A training cohort of 519 participants yielded an optimal diagnostic cutoff of serum AKR1B10 at 267.9 pg/mL. When ROC curve was plotted for HCC versus all controls (HC + BLT + CHB + LC), serum AKR1B10 had diagnostic parameters of the area under the curve (AUC) 0.896, sensitivity 72.7%, and specificity 95.7%, which were better than AFP with AUC 0.816, sensitivity 65.1%, and specificity 88.9%. Impressively, AKR1B10 showed promising diagnostic potential in early-stage HCC and AFP-negative HCC. Combination of AKR1B10 with AFP increased diagnostic accuracy for HCC compared with AKR1B10 or AFP alone. A validation cohort of 522 participants confirmed these findings. An independent cohort of 68 patients with HCC who were followed up showed that serum AKR1B10 dramatically decreased 1 day after operation and was nearly back to normal 3 days after operation. Conclusion: AKR1B10 is a potent serum marker for detection of HCC and early-stage HCC, with better diagnostic performance than AFP.

Helicobacter hepaticus infection induces chronic hepatitis and fibrosis in male BALB/c mice via the activation of NF-κB, Stat3, and MAPK signaling pathways.

BACKGROUND: It has been documented that Helicobacter hepaticus (H hepaticus) infection is linked to chronic hepatitis and liver cancer. However, our understanding of the molecular mechanisms underlying progression of the H hepaticus-induced hepatic inflammation to cellular hepatocarcinoma is still limited. MATERIALS AND METHODS: In our study, male BALB/c mice were infected by H hepaticus for 8, 12, 16, 20, and 24 weeks. Histopathology, H hepaticus colonization dynamics, select signaling pathways, and expression of key inflammatory cytokines in the liver were examined. RESULTS: We found that H hepaticus was detectible in feces of mice at 7 days postinfection (DPI) by PCR, but it was not detected in the livers by PCR until 8 weeks postinfection (WPI). In addition, abundance of colonic and hepatic H hepaticus was progressively increased over the infection duration. H hepaticus-induced hepatic inflammation and fibrosis were aggravated over the infection duration, and necrosis or cirrhosis developed in the infected liver at 24 WPI H hepaticus infection increased levels of alanine aminotransferase and aspartate aminotransferase. Moreover, mRNA levels of Il-6 and Tnf-α were significantly elevated in the livers of H hepaticus-infected mice compared to uninfected control from 8 WPI to 24 WPI. Furthermore, Stat3, nuclear factor-κB (p65), and MAPK (Erk1/2 and p38) were activated by H hepaticus infection. CONCLUSIONS: These data demonstrated that male BALB/c mice can be used as a new mouse model of H hepaticus-induced liver diseases and that the H hepaticus-induced liver injury is triggered by NF-κB, Jak-Stat, and MAPK signaling pathways.

The influence of exposure to Toxoplasma Gondii on host lipid metabolism.

BACKGROUND: Mounting evidence suggested a complex correlation between host lipid metabolism and Toxoplasma gondii (T. gondii) infection. However, the inherent association between T. gondii infection and host lipid state remains elusive either in mice or in human hosts. METHODS: Through a study in a sample of 1045 healthy participants from eastern China, we determined the association of T. gondii infection and host lipid levels using serological methods. We then examined the host lipid levels in C57BL/6 J mice at both acute and chronic T. gondii infection stages (for a period up to 36 weeks post infection). RESULTS: In our case-control study, T. gondii seropositive individuals had higher low-density lipoproteins (LDL) (P = 0.0043) and total cholesterol (TC) (P = 0.0134) levels compared to seronegative individuals. Furthermore, individuals with LDL (OR = 3.25; 95% CI:1.60-6.61) and TC (OR = 2.10; 95% CI:1.22-3.63) levels above the upper limit of normal range had higher odds ratio to be T. gondii IgG positive. Consistently, in vivo data revealed that a significantly increased LDL level was first observed at early acute stage but plateaued to later time (chronic infection with T. gondii). CONCLUSIONS: In both healthy population and T. gondii-infected mice, seropositive individuals had higher LDL level. Individuals with positive T. gondii IgG had more odds of being with LDL and TC abnormality. Latent T. gondii infection is common worldwide, potential medical interventions to host lipid metabolism may be a breakthrough point to the prevention and control of this parasite infection.

PKCλ/ι regulates Th17 differentiation and house dust mite-induced allergic airway inflammation.

Asthma is a chronic airway inflammation in which Th2 and Th17 cells play critical roles in its pathogenesis. We have reported that atypical protein kinase (PKC) λ/ι is a new regulator for Th2 differentiation and function. However, the role of PKCλ/ι for Th17 cells remains elusive. In this study, we explored the effect of PKCλ/ι on Th17 cells in the context of ex vivo cell culture systems and an in vivo murine model of allergic airway inflammation with the use of activated T cell-specific conditional PKCλ/ι-deficient mice. Our findings indicate that PKCλ/ι regulates Th17 cells. The secretion of Th17 effector cytokines, including IL-17, IL-21 and IL-22, were inhibited from PKCλ/ι-deficient T cells under non-skewing or Th17-skewing culture conditions. Moreover, the impaired Th17 differentiation and function by the PKCλ/ι-deficiency was associated with the downregulation of Stat3 and Rorγt, key Th17 transcription factors. We developed a model of Th17 and neutrophil-involved allergic airway inflammation by intratracheal inoculation of house dust mites. PKCλ/ι-deficiency significantly inhibited airway inflammations. The infiltrating cells in the lungs and bronchoalveolar lavage fluids were significantly reduced in conditional PKCλ/ι-deficient mice. Th17 effector cytokines were reduced in the bronchoalveolar lavage fluids and lungs at protein and mRNA levels. Thus, PKCλ/ι emerges as a critical regulator of Th17 differentiation and allergic airway hyperresponsiveness.

Recurrence of diet-treated gestational diabetes in primiparous women in northern Zhejiang, China: Epidemiology, risk factors and implications.

AIM: This study sought to determine the rate of recurrence of gestational diabetes mellitus (GDM) recurrence during the second pregnancies of women who were diagnosed with GDM during their first pregnancies, to identify risk factors associated with the probability of such recurrence and to evaluate the influence of GDM recurrence on pregnancy outcomes in north Zhejiang, China, after the recent adjustment to the nation's childbirth policy. METHODS: A retrospective longitudinal study was performed in north Zhejiang, China (at Jiaxing Maternal and Child Health Hospital). A total of 128 women who delivered two sequential live singleton infants and were diagnosed with diet-treated GDM during their first pregnancies were included. RESULTS: According to the 2013 World Health Organization diagnostic criteria for diabetes during pregnancy, the prevalence of gestational diabetes was 11.02% in northern Zhejiang. The recurrence rate of GDM in northern Zhejiang was 43.75% (56/128). The age at second pregnancy, weight gain during pregnancy, interpregnancy interval and macrosomia during the index pregnancy were risk factors for GDM recurrence. Among those women with recurrent GDM, GDM developed earlier and caesarean section was more frequently required during the second pregnancy; in addition, the second pregnancy was associated with more premature and low birthweight infants but less macrosomia. CONCLUSION: The recurrence rate of GDM is high in northern Zhejiang. Glucose monitoring and management are needed during subsequent pregnancies for patients who previously presented with GDM to improve maternal and fetal outcomes.

Investigations on the interactions between naphthalimide-based anti-tumor drugs and human serum albumin by spectroscopic and molecular modeling methods.

The interactions between the three kinds of naphthalimide-based anti-tumor drugs (NADA, NADB, NADC) and human serum albumin (HSA) under simulated physiological conditions were investigated by fluorescence spectroscopy, circular dichroism spectroscopy and molecular modeling. The results of the fluorescence quenching spectroscopy showed that the quenching mechanisms for different drugs were static and their affinity was in a descending order of NADA > NADB > NADC. The relative thermodynamic parameters indicated that hydrophobic force was the predominant intermolecular force in the binding of NAD to HSA, while van der Waals interactions and hydrogen bonds could not be ignored. The results of site marker competitive experiment confirmed that the binding site of HSA primarily took place in site I. Furthermore, the molecular modeling study was consistent with these results. The study of circular dichroism spectra demonstrated that the presence of NADs decreased the α-helical content of HSA and induced the change of the secondary structure of HSA.

[Effect of Toxoplasma gondii Infection in Female Mice on Dopamine Level in the Brain of the Male Offspring].

OBJECTIVE: To investigate the effect of Toxoplasma gondii infection in female mice on dopamine level in the brain of male offspring. METHODS: Thirty-six ICR female mice were randomly divided into control group and infection group, 18 mice in each group. Each mouse in infection group was orally infected with 10 cysts of T. gondii Prugniaud strain. On the 90th day after infection, the infected female mice were mated with normal male ICR mice at 1:1 ratio. On the 20th day of pregnancy, 2 mice in each group were delivered for fetal mice by cesarean section, and the brain of male fetal mice (n = 6) in each group were collected. On the 14th and 63rd day after birth, 6 male offspring mice in each group were sacrificed, and the brain were collected. Dopamine levels in the cortex, cerebellum, hippocampus, and striatum were analyzed by high-performance liquid chromatography-electrochemical detection (HPLC-ECD). RESULTS: Three mice in infection group died during the experiment, and 6 out of 15 female mice mated successfully. The number of fetal mice and F1 generation mice in infection group was 12 (male: 7) and 21 (male: 15), respectively. All the mice in control group mated successfully. The number of fetal mice and F1 generation mice was 23 (male: 12) and 179 (male: 92), respectively. The dopamine level in the cerebellum of fetal mice of infection group and control group was (413.25 ± 21.78) ng/g and (346.30 ± 51.83) ng/g, respectively (P < 0.01). No significant difference was found in dopamine content in the cortex between the two groups (P > 0.05). Compared with the control group, on the 14th day and 63rd day after birth, the dopamine content in cortical areas [(462.50 ± 24.80) ng/g and (1215.77 ± 113.64) ng/g], cerebellum area [(271.55 ± 26.19) ng/g and (1328.82 ± 39.62) ng/g], hippocampus area [(225.78 ± 24.17) ng/g and (1322.70 ± 58.34) ng/g], and striatum area [(455.23 ± 61.53) ng/g and (991.32 ± 54.31) ng/g] of the male offspring in infection group were significantly higher than that of the control (P < 0.05, P < 0.01). CONCLUSION: T. gondii infection in female mice causes an increase of dopamine level in the brain of F1 generation male mice.

Stem cell therapy for the treatment of parasitic infections: is it far away?

Stem cell therapy is an interventional treatment that introduces new cells into damaged tissues, which help in treating many diseases and injuries. It has been proved that stem cell therapy is effective for the treatment of cancers, diabetes mellitus, Parkinson's disease, Huntington's disease, cardiovascular diseases, neurological disorders, and many other diseases. Recently, stem cell therapy has been introduced to treat parasitic infections. The culture supernatant of mesenchymal stem cells (MSCs) is found to inhibit activation and proliferation of macrophages induced by the soluble egg antigen of Schistosoma japonicum, and MSC treatment relieves S. japonicum-induced liver injury and fibrosis in mouse models. In addition, transplantation of MSCs into naïve mice is able to confer host resistance against malaria, and MSCs are reported to play an important role in host protective immune responses against malaria by modulating regulatory T cells. In mouse models of Chagas disease, bone marrow mononuclear cell has been shown effective in reducing inflammation and fibrosis in mice infected with Trypanosoma cruzi, and transplantation of the bone marrow mononuclear cells prevents and reverses the right ventricular dilatation induced by T. cruzi infection in mice. Preliminary clinical trials demonstrate that transplantation of bone marrow derived-cells may become an important therapeutic modality in the management of end-stage heart diseases associated with Chagas disease. Based on these exciting results, it is considered by stating that it is firmly believed that, within the next few years, we will be able to find the best animal models and the appropriate stem cell type, stem cell number, injection route, and disease state that will result in possible benefits for the patients with parasitic infections, and stem cell therapy, although at an initial stage currently, will become a real therapeutic option for parasitic diseases.

[Dynamic changes of sciatic nerve conduction velocity in rats infected with Toxoplasma gondii].

OBJECTIVE: To observe the dynamic changes of sciatic nerve conduction velocity of Toxoplasma gondii-infected rats at different time points. METHODS: Twenty SD rats were randomly divided into control group and Toxoplasma gondii infection group. Rats in T. gondii infection group were intraperitoneal injected with 4x10(7) T. gondii tachyzoites, while those in control group were given equivalent normal saline. Motor and sensory nerve conduction velocities (MNCV, SNCV) in sciatic nerve were measured by Medtronic Keypoint4 Workstation electromyography at pre-infection, and 2, 4, 8, 12 months post-infection. RESULTS: Within two months after infection, there was no difference in SNCV and MNCV between control group and infection group (P>0.05). From 4 months after T. gondii injection, infected rats began to show the slowness of SNCV and MNCV, which progressed with the course of infection. At 4, 8, and 12 months after infection, SNCV and MNCV of infection group were (35.26±3.02) and (25.94±3.20) m/s, (33.57±2.27) and (22.75±2.31) m/s, and (32.38±2.38) and (22.03±2.08) m/s, respectively. Compared with control group, SNCV and MNCV of infection group reduced by (7.47±2.11)% and (12.57±1.89)%, (8.92±2.64)% and (13.72±2.65)%, and (12.18±1.94)% and (15.46±2.37)%, respectively (P<0.05). CONCLUSION: From 4 months after infection, Toxoplasma gondii-infected rats show a slowness of motor and sensory nerve conduction velocities in sciatic nerve.

Dysregulated Glucuronidation of Bilirubin Exacerbates Liver Inflammation and Fibrosis in Schistosomiasis Japonica through the NF-κB Signaling Pathway.

Hepatic fibrosis is an important pathological manifestation of chronic schistosome infection. Patients with advanced schistosomiasis show varying degrees of abnormalities in liver fibrosis indicators and bilirubin metabolism. However, the relationship between hepatic fibrosis in schistosomiasis and dysregulated bilirubin metabolism remains unclear. In this study, we observed a positive correlation between total bilirubin levels and the levels of ALT, AST, LN, and CIV in patients with advanced schistosomiasis. Additionally, we established mouse models at different time points following S. japonicum infection. As the infection time increased, liver fibrosis escalated, while liver UGT1A1 consistently exhibited a low expression, indicating impaired glucuronidation of bilirubin metabolism in mice. In vitro experiments suggested that SEA may be a key inhibitor of hepatic UGT1A1 expression after schistosome infection. Furthermore, a high concentration of bilirubin activated the NF-κB signaling pathway in L-O2 cells in vitro. These findings suggested that the dysregulated glucuronidation of bilirubin caused by S. japonicum infection may play a significant role in schistosomiasis liver fibrosis through the NF-κB signaling pathway.

[Proteomic study of the hippocampus tissue from rats with chronic Toxoplasma gondii infection].

OBJECTIVE: To observe the proteome changes in the hippocampus tissue of rats with chronic Toxoplasma gondii infection. METHODS: Six male SD rats were randomly divided into control group and infection group. Each rat in infection group was intraperitoneally injected with 4 x 10(7) purified T. gondii tachyzoites. Rats in the control group received equivalent volumes of sterile normal saline. At the fifth day post-infection, blood samples were taken from the lateral tail vein and Ciemsa staining of blood cells was performed to find Toxoplasma gondii. Rats were dissected at the 10th week post-infection, total protein in the hippocampus was separated by using two-dimensional gel electrophoresis (2-DE). After Coomassie blue staining, the Image Analysis software was used to select and separate proteins on the gel. Matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF-MS) was used for peptide mass fingerprint PMF). Proteins were identified by using Mascot software to search the MSDB and SwissProt databases. RESULTS: Microscopy examination of blood smears confirmed that the rats in infection group were all infected by 11 gondii. The number of protein spots of rats from infection group and control group was 311 +/- 19 and 327 +/- 13 respectively. Compared with the control group, 5 protein spots disappeared, 4 protein spots were up-regulated and 7 were down-regulated in the infection group. The 9 differentially expressed protein spots were identified by MALDL-TOF-MS: phosphoglycerate kinase 1, similar to alpha-enolase, glutamine synthetase, creatine kinase, creatine kinase B-type, ATP synthase, aconitase 2, mitochondrial precursor, actin and an unnamed protein. The first three proteins were up-regulated and the other five proteins were down-regulated in infection group. CONCLUSION: Nine differential expression proteins are found from the hippocampus tissue in rats chronically infected with T. gondii and normal SD rats.

Thermodynamic and Kinetic Effects of Quaternary Ammonium and Phosphonium Ionic Liquids on CO2 Hydrate Formation.

The paper elaborates the effects of ionic liquids (ILs) on the phase equilibrium temperature, induction time, gas consumption, gas consumption rate, and water to hydrate conversion in the presence of 0.25, 0.63, 0.95, 1.25, 3.75, 6.25, and 10.00 wt % ethyltributylphosphonium hexafluorophosphate ([P2 4 4 4][PF6]), tributylhexylphosphonium hexafluorophosphate ([P6 4 4 4][PF6]), tetraethylammonium bromide ([N2 2 2 2]Br), tetraethylammonium bistrifluoromethanesulfonimide ([N2 2 2 2][NTf2]), and tetraethylammonium hexafluorophosphate ([N2 2 2 2][PF6]) under a pressure of 2 MPa. The results indicate that all five ILs could increase CO2 consumption and enhance the water to hydrate conversion. Compared with the pure water system, [P2 4 4 4][PF6] and [P6 4 4 4][PF6] shifted the phase equilibrium temperature of CO2 hydrates to a slightly higher temperature with reduced induction times by boosting CO2 hydrate nucleation, showing the dual function promotion effects. In contrast, [N2 2 2 2]Br, [N2 2 2 2][NTf2], and [N2 2 2 2][PF6] shifted the phase equilibrium temperature of CO2 hydrates to a lower temperature and prolonged the induction time by slowing down CO2 hydrate nucleation. The inhibition effects of anions on CO2 hydrates follow an order of Br- > [NTf2]- > [PF6]-. Besides, the density functional theory and molecular dynamic calculations were conducted to explain the inconsistent influences of [N2 2 2 2]Br and [N4 4 4 4]Br on CO2 hydrate formation. It was found that the anion-cation interaction of [N2 2 2 2]Br was stronger than that of [N4 4 4 4]Br, and Br- in [N2 2 2 2]Br is less likely to participate in the formation of hydrate cages in the [N2 2 2 2]Br + H2O + CO2 system according to the intermolecular anion-water, anion-CO2, and water-water radial distribution function in [N2 2 2 2]Br + H2O + CO2 and [N4 4 4 4]Br + H2O + CO2 systems.

Blocking BAFF Alleviates Hepatic Fibrosis in Schistosoma japonicum-Infected Mice.

Schistosomiasis is an immunopathogenic disease characterized by egg granuloma and fibrosis. The hepatic fibrosis of schistosomiasis is caused by the coordinated action of local immune cells, liver-resident cells and related cytokines around the eggs of the liver. B-cell-activating factor (BAFF), expressed in many cells, is an essential factor for promoting the survival, differentiation, and maturation of cells. The overexpression of BAFF is closely related to many autoimmune diseases and fibrosis, but has not been reported to play a role in liver fibrosis caused by schistosomiasis. In the study, we found that, during Schistosoma japonicum (S. japonicum) infection in mice, the level of BAFF and its receptor BAFF-R progressively increased, then decreased with the extension of infection time, which was consistent with the progression of hepatic granuloma and fibrosis. Anti-BAFF treatment attenuated the histopathological damage in the liver of infected mice. The average areas of individual granulomas and liver fibrosis in anti-BAFF treatment mice were significantly lower than those in control mice, respectively. Anti-BAFF treatment increased the IL-10, decreased IL-4, IL-6, IL-17A, TGF-ß, and downregulated the antibody level against S. japonicum antigens. These results suggested that BAFF acts a strong player in the immunopathology of schistosomiasis. Anti-BAFF treatment may influence Th2 and Th17 responses, and reduce the inflammatory reaction and fibrosis of schistosomiasis liver egg granuloma. It is suggested that BAFF might be a prospective target for the development of new methods to treat schistosomiasis liver fibrosis.

Increased production and anti-senescence activity of exopolysaccharides in Ganoderma lingzhi by co-overexpression of ß-1,3-glucan synthase and UDP-glucose pyrophosphorylase.

A ß-1,3-glucan synthase gene (gls) was cloned and overexpressed in Ganoderma lingzhi. The content of intracellular polysaccharides (IPS) in G. lingzhi overexpressing gls was 22.36 mg/100 mg dry weight (DW), 19 % higher than those in the wild-type (WT) strain. Overexpression of gls did not affect the expression of the phosphoglucomutase gene and the UDP-glucose pyrophosphorylase gene (ugp) in the polysaccharide biosynthesis. The gls and ugp were then simultaneously overexpressed in G. lingzhi for the first time. The combined overexpression of these two genes increased the IPS content and exopolysaccharides (EPS) production to a greater extent than the overexpression of gls independently. The maximum IPS content of the overexpressed strain was 24.61 mg/100 mg, and the maximum EPS production was 1.55 g/L, 1.31- and 1.50-fold higher than that in the WT strain, respectively. Moreover, the major EPS fractions from the overexpression strain contained more glucose (86.7 % and 72.5 %) than those from the WT strain (78.2 % and 62.9 %). Furthermore, the major fraction G+U-0.1 from the overexpression strain exhibited stronger antioxidant and anti-senescence activities than the WT-0.1 fraction from the WT strain. These findings will aid in the hyperproduction and application of Ganoderma polysaccharides and facilitate our understanding of mushroom polysaccharide biosynthesis.

Study on Thermal Risk of Coal-Based Activated Carbon after Adsorbing Acetone, Cyclohexane, and Butyl Acetate.

Combustion and explosion accidents of the mixture may occur after the adsorption of volatile organic compounds (VOCs) by coal-based activated carbon (CBAC). It is of great significance to explore the oxidation and combustion performance of CBAC before and after adsorbing VOCs in order to prevent the reoccurrence of fire and explosion. Based on the CBAC sample commonly used in industrial production, three types of CBAC samples after adsorbing VOCs, i.e., acetone, cyclohexane, and butyl acetate, were prepared. The oxidation and combustion characteristics of the samples before and after adsorbing VOCs are measured and analyzed by thermal analyzer and cone calorimeter. Thermal analysis results indicate that during the oxidation process, the VOCs in the adsorbed samples will burn in the early stage, generating amounts of heat which may accelerate the oxidation and combustion of CBAC. According to the combustion performance experiments by cone calorimeter, it is also found that the combustion rate of CBAC after adsorbing VOCs is significantly enhanced. The time to ignition is shortened, the heat release rate becomes larger, and the time to reach the peak of heat release rate is significantly moved forward. In addition, the CO yield of the adsorbed sample is significantly improved. In general, VOC adsorption in CBAC can promote oxidation reactions and may result in an enhanced combustibility of CBAC.

Study of the Microstructure of Coal at Different Temperatures and Quantitative Fractal Characterization.

In order to understand the influence of underground coal fires on coal fractures and pores, mercury intrusion porosimetry (MIP) and scanning electron microscopy (SEM) are combined to study the development of coal pore and fracture under high-temperature treatment and calculate the fractal dimension to analyze the relationship between the development of coal pore and fracture and the fractal dimension. The results show that the volume of pores and fractures of the coal sample (C200) treated at 200 °C (0.1715 mL/g) is greater than that of the coal sample (C400) treated at 400 °C (0.1209 mL/g), and both are greater than the original coal sample (RC) (0.1135 mL/g). The volume increase is mainly due to mesopores and macropores, and the proportions of mesopores and macropores in C200 were 70.15 and 59.97% in C400. The MIP fractal dimension shows a decreasing trend with the increase of temperature, and the connectivity of coal samples improved with the increase of temperature. The changes in volume and three-dimensional fractal dimension of C200 and C400 showed the opposite trend and are related to the different stress of coal matrix at different temperatures. The experimental SEM images confirm that the connectivity of coal fractures and pores improves with the increase of temperature. Based on the SEM experiment, the larger the fractal dimension, the more complex the surface is. The SEM surface fractal dimensions indicate that the surface fractal dimension of C200 is the smallest and that of C400 is the largest, which is consistent with the observations made by SEM. The combination of the two fractal dimensions is used to characterize the self-similarity of coal using the fractal dimension difference. When the temperature increased to 200 °C, the unordered expansion of the coal sample resulted in the largest fractal dimension difference and the lowest self-similarity. When heated to 400 °C, the fractal dimension difference of the coal sample is the smallest, and the microstructure of coal shows a regular groove-like development.

Deficiency of PKCλ/ι alleviates the liver pathologic impairment of Schistosoma japonicum infection by thwarting Th2 response.

BACKGROUND: The activation of immune response driven by the eggs of Schistosoma japonicum and the subsequent secretions is the culprit behind granulomatous inflammation and liver fibrosis. Evidence suggests that PKCλ/ι participates in a variety of physiological and pathological processes, including the regulation of metabolism, growth, proliferation and differentiation of cells. However, the role of PKCλ/ι in liver disease caused by Schistosoma japonicum remains unclear. METHODS: In the present study, we observe the pathological changes of egg-induced granulomatous inflammation and fibrosis in the liver of mice infected by Schistosoma japonicum by using conditional PKCλ/ι-knockout mice and wild-type control. Immune cytokines and fibrogenic factors were analyzed by performing flow cytometry and real-time fluorescence quantitative PCR. RESULTS: The results of H&E and Masson staining show that the degree of granulomatous lesions and fibrosis in the liver of the infected PKCλ/ι-knockout mice was significantly reduced compared with those of the infected wild-type mice. The mean area of single granuloma and hepatic fibrosis in the PKCλ/ι-knockout mice was significantly lower than that of the wild-type mice (85,295.10 ± 5399.30 µm2 vs. 1,433,702.04 ± 16,294.01 µm2, P < 0.001; 93,778.20 ± 8949.05 µm2 vs. 163,103.01 ± 11,103.20 µm2, P < 0.001), respectively. Serological analysis showed that the ALT content was significantly reduced in the infected knockout mice compared with infected wild-type mice. RT-PCR analysis showed that IL-4 content in knockout mice was significantly increased after Schistosoma japonicum infection, yet the increase was less than that in infected wild-type mice (P < 0.05). PKCλ/ι deficiency led to reduced expression of fibrosis-related factors, including TGF-ß1, Col-1, Col-3, α-SMA and liver DAMP factor HMGB1. Flow cytometry analysis showed that the increasing percentage of Th2 cells, which mainly secrete IL-4 cytokines in spleen cells, was significantly lower in PKCλ/ι-deficient mice compared with wild-type mice after infection (P < 0.05). CONCLUSIONS: Our data demonstrate that PKCλ/ι deficiency alleviating granulomatous inflammation and fibrosis in the liver of mice with S. japonicum infection by downregulating Th2 immune response is the potential molecular mechanism behind the role of PKCλ/ι in schistosomiasis.