Experimental Study on Heat Conduction and Water Migration of Composite Bentonite Samples.

The joints of buffer material composite blocks as potential weak parts in the engineering barrier system of a high-level radioactive waste (HLW) repository must be studied in depth. Therefore, a laboratory experiment device suitable for unsaturated composite bentonite samples was developed. The evolution of temperature and volumetric water content at different locations of Gaomiaozi (GMZ) composite bentonite samples with time before and after simulated water inflow was measured by the experiment device. According to the experimental results, the thermal conductivity and hydraulic conductivity of the joint location after healing of the composite bentonite samples were obtained. The experimental results show that the change in the internal temperature of the composite bentonite samples is mainly affected by the temperature boundary and that the change in the internal water has little effect on it. In a short period of time, the loading of hydraulic boundary conditions only makes the volumetric water content of the soil near the hydraulic boundary increase significantly but has little effect on other locations. And, affected by the temperature boundary, the volumetric water content of the soil near the temperature boundary gradually decreases with time. The process of hydration swelling of the composite bentonite sample is accompanied by the adjustment of stress. The composite bentonite samples are continuously squeezed to the joint area after hydration swelling, the whole composite samples are generally homogenized, and the joints between the composite bentonite samples tend to heal. The thermal conductivity and permeability of the joint location after healing can meet the requirements of the engineering barrier of the HLW repository.

Tumor-Associated Macrophage-Derived TGF-ß1 Activates GLI2 via the Smad2/3 Signaling Pathway to Affect Cisplatin Resistance in Lung Adenocarcinoma.

BACKGROUND: Transforming growth factor-ß1 (TGF-ß1) is an immunosuppressive cytokine that is highly expressed in the tumor microenvironment (TME) of lung adenocarcinoma (LUAD). TGF-ß1 plays important roles in regulating tumor metastasis and chemotherapy resistance. However, the specific molecular mechanisms by which TGF-ß1 regulates cisplatin resistance in the TAM of LUAD remain unclear. MATERIALS AND METHODS: THP-1 induced macrophages were co-cultured with A549 and H1975 cells, and subsequently transfected with silencing TGF-ß1 (siTGF-ß1), GLI2 (siGLI2), a GLI2 overexpression plasmid, and their negative controls. Cellular activity was measured by CCK-8 and colony formation assays. Cell apoptosis was evaluated by flow cytometry and TUNEL staining. Transwell assays were performed to assess cell migration and invasion capabilities. The levels of Smad2/3, GLI2, cyclin D, and cyclin E expression were evaluated by qPCR, western blotting, and immunofluorescence methods. TGF-ß1 levels were determined by ELISA. RESULTS: Macrophages suppressed the apoptosis and promoted the migration and invasion of LUAD cells. TAM siTGF-ß1 downregulated the Smad2/3 signaling pathways and GLI2 expression, deceased cell proliferation, and promoted apoptosis. SiGLI2 increased apoptosis and decreased the proliferation of LUAD cell lines. GLI2 decreased cisplatin resistance in LUAD cells. CONCLUSION: High expression of TGF-ß1 in the TAM positively activates GLI2 expression via the Smad2/3 pathway, which subsequently regulates cyclin D and cyclin E expression, and promotes the cisplatin resistance of LUAD.

Carvedilol to prevent hepatic decompensation of cirrhosis in patients with clinically significant portal hypertension stratified by new non-invasive model (CHESS2306).

Background & Aims: Non-invasive models stratifying clinically significant portal hypertension (CSPH) are limited. Herein, we developed a new non-invasive model for predicting CSPH in patients with compensated cirrhosis and investigated whether carvedilol can prevent hepatic decompensation in patients with high-risk CSPH stratified using the new model. Methods: Non-invasive risk factors of CSPH were identified via systematic review and meta-analysis of studies involving patients with hepatic venous pressure gradient (HVPG). A new non-invasive model was validated for various performance aspects in three cohorts, i.e., a multicenter HVPG cohort, a follow-up cohort, and a carvedilol-treating cohort. Results: In the meta-analysis with six studies (n = 819), liver stiffness measurement and platelet count were identified as independent risk factors for CSPH and were used to develop the new "CSPH risk" model. In the HVPG cohort (n = 151), the new model accurately predicted CSPH with cutoff values of 0 and -0.68 for ruling in and out CSPH, respectively. In the follow-up cohort (n = 1,102), the cumulative incidences of decompensation events significantly differed using the cutoff values of <-0.68 (low-risk), -0.68 to 0 (medium-risk), and >0 (high-risk). In the carvedilol-treated cohort, patients with high-risk CSPH treated with carvedilol (n = 81) had lower rates of decompensation events than non-selective beta-blockers untreated patients with high-risk CSPH (n = 613 before propensity score matching [PSM], n = 162 after PSM). Conclusions: Treatment with carvedilol significantly reduces the risk of hepatic decompensation in patients with high-risk CSPH stratified by the new model.

A preliminary study on the evaluation of photodynamic diagnosis for acne inflammatory skin lesions.

OBJECTIVE: This study aimed to observe the fluorescence characteristics of acne inflammatory skin lesions in 5-aminolevulinic acid-based photodynamic diagnosis (ALA-PDD), and discuss the viability of using ALA-PDD to evaluate acne inflammatory skin lesions and explore the advantages of predicting subclinical skin lesions. METHODS: The OBSERV facial skin detector collected photographs of 20 patients before and after optical intra-tissue fiber irradiation photodynamic therapy (OFI-ALA-PDT) in both ALA-PDD and white light patterns. The patients were treated once a week for four consecutive weeks in order to analyze the correlation between the two patterns in recognizing inflammatory skin lesions. RESULTS: Before and after treatment, there was no significant difference between the two patterns for recognizing acne inflammatory skin lesions (p > 0.05). Both patterns demonstrated a strong correlation (r > 0.90) for the recognition of various types of inflammatory skin lesions at different treatment stages. CONCLUSION: ALA-PDD is a feasible method for evaluating acne inflammatory lesions, guiding treatment and judging efficacy. It has advantages in predicting subclinical skin lesions and deserves further study.

Global profiling of AMG510 modified proteins identified tumor suppressor KEAP1 as an off-target.

KRAS inhibitor AMG510 covalently modifies the G12C residue and inactivates the KRAS/G12C function. Because there are many reactive cysteines in the proteome, it is important to characterize AMG510 on-target modification and off-targets. Here, we presented a streamlined workflow to measure abundant AMG510 modified peptides including that of KRAS/G12C by direct profiling, and a pan-AMG510 antibody peptide IP workflow to profile less abundant AMG510 off-targets. We identified over 300 off-target sites with three distinct kinetic patterns, expanding the AMG510 modified proteome involved in the nucleocytoplasmic transport, response to oxidative stress, adaptive immune system, and glycolysis. We found that AMG510 covalently modified cys339 of ALDOA and inhibited its enzyme activity. Moreover, AMG510 modified KEAP1 cys288 and induced NRF2 accumulation in the nuclear of NSCLC cells independent of KRAS/G12C mutation. Our study provides a comprehensive resource of protein off-targets of AMG510 and elucidates potential toxicological sideeffects for this covalent KRASG12C inhibitor.

LncRNA AC098934 promotes proliferation and invasion in lung adenocarcinoma cells by combining METTL3 and m6A modifications.

Background: Long non-coding RNA (lncRNA) regulates the tumorigenesis as well as the development of lung adenocarcinoma (LUAD), which is one of the high-mortality cancers. We explored the influence of lncRNA AC098934 on the malignant biological behavior of LUAD and potential underlying molecular mechanisms. Methods: The expression level of AC098934 in either the LUAD or the normal tissues was identified in the TCGA database. Two AC098934 knockdown siRNAs were infected into cells of LUAD, including A549 as well as H1299 cells, using the lentivirus. Real-time Quantitative polymerase chain reaction (QPCR) helped to determine the knockdown efficiency of AC098934. CCK-8, cell cloning, wound healing combined with transwell assays tested the role of AC098934 in the cell proliferation, migration as well as the invasion. Tumor formation experiment in nude mice subcutaneously confirmed the promoting effect of AC098934 in vivo. In addition, combinations of METTL3 and AC098934, as well as m6A and AC098934 were identified through the RIP assay. Results: Compared to the normal tissues, AC098934 was more highly expressed in LUAD tissues. After AC098934 was knocked down by siRNA, the proliferation, invasion, migration as well as tumorigenesis abilities of both A549 and H1299 cells were reduced. Mechanistically, AC098934 could bind to the m6A antibody and METTL3 protein. METTL3 overexpression promoted the m6A modification on AC098934, thereby increasing the interaction of m6A modification. Conclusion: The highly expressed lncRNA AC098934 in LUAD facilitates the cell proliferation as well as invasion either in vitro or in vivo. METTL3 binds, furthermore, modulates the m6A modification of AC098934. Our research revealed a new molecular mechanism, through which AC098934 promoted the malignant behavior of LUAD tumors under the m6A modification induced by METTL3. This indicates that AC098934 is possible to be a promising biomarker as well as a therapeutic target for the patients with LUAD.

Pyroptosis-Related Gene Signature Is a Novel Prognostic Biomarker for Sarcoma Patients.

Sarcoma is a rare and an extremely aggressive form of cancer that originates from mesenchymal cells. Pyroptosis exerts a dual effect on tumours by inhibiting tumour cell proliferation while creating a microenvironment suitable for tumour cell development and proliferation. However, the significance of pyroptosis-related gene (PRG) expression in sarcoma has not yet been evaluated. Here, we conduct a retrospective analysis to examine PRG expression in 256 sarcoma samples from The Cancer Genome Atlas database. We identified the PRGs that had a significant correlation with overall patient survival in sarcoma by performing a univariate Cox regression analysis. Subsequently, we conducted a LASSO regression analysis and created a risk model for a six-PRG signature. As indicated from the Kaplan-Meier analysis, this signature revealed a significant difference between high- and low-risk sarcoma patients. A receiver operating characteristic curve analysis confirmed that this signature could predict overall patient survival in sarcoma patients with high sensitivity and specificity. Gene ontology annotation and Kyoto Encyclopaedia of Genes and Genomes pathway enrichment analyses revealed that five independent PRGs were closely associated with increased immune activity. Moreover, we also deciphered that increased number of immune cells infiltrated the tumour microenvironment in sarcoma. In brief, the PRG signature can effectively act as novel prognostic biomarker for sarcoma patients and is associated with the tumour immune microenvironment.

Acupuncture paired with herbal medicine for prediabetes: study protocol for a randomized controlled trial.

BACKGROUND: Type-2 diabetes has become a major disease and is known to seriously impair people's health worldwide. Prediabetes includes impaired fasting glucose (IFG) and impaired glucose tolerance (IGT) and is the most critical period for preventing type-2 diabetes, as it can be identified and reversed. Studies in the past decade have indicated that acupuncture and Chinese herbal medicine may be beneficial for treating prediabetes. However, a randomized controlled trial (RCT) should be conducted to obtain more clinical evidence on this topic. METHODS/DESIGN: An RCT will be implemented in this study, using a72-week study period (24 weeks for the intervention and 48 weeks for follow-up). Participants will be recruited from the Fifth Affiliated Hospital of Guangzhou Medical University in China. Eighty participants will be randomized to the treatment group (acupuncture plus herbal medicine and health education) or the control group (health education only), 40 participants in each. People included in this study must have been diagnosed with prediabetes using Western medicine criteria. The endpoint indices include the incidence of diabetes mellitus and the reversion rate. The primary outcome is fasting plasma glucose (FPG) level, 2-h plasma glucose (2-hPG) level after a 75-g oral glucose tolerance test (OGTT), and glycosylated hemoglobin (HbA1c) level. Secondary outcomes include the following: Body Mass Index (BMI); hemorheology, including shear rates of whole-blood viscosity and plasma viscosity. Safety indices include hepatic (ALT, AST) and renal function (BUN, Cr) and records of adverse events, including diarrhoea, colds, pharyngitis, and sleep disorders. Quality control will be implemented, including quality control of the laboratory, researchers, participants, investigational drugs, data and documents, occurrence of bias, supervision, among others, according to uniform standard operating procedures (SOPs) which have been established by the Good Clinical Practice (GCP) office of the Fifth Affiliated Hospital of Guangzhou Medical University. DISCUSSION: The aim of this study is to evaluate the efficacy and safety of acupuncture paired with herbal medicine for the treatment of patients with prediabetes. TRIAL REGISTRATION: Chinese clinical trials register ChiCTR-INR-16008891 . Registered on 23 July 2016.

[Analysis in 13 315 newborns hearing screening].

OBJECTIVE: Explore the model of universal NICU newborns' hearing screening in high-risk neonates, preliminary understanding factor of hearing damage. METHOD: Transient evoked otoacoustic emissions (TEOAE) and automatic auditory brainstem response (AABR) were used to detect newborns' hearing in 13 315 objects, that is newborns' hearing screening in NICU with TEOAE test who not pass, 42 days after will use AABR rescreening. Children's Hearing Center of Guangxi Child Health Hospital will diagnose the newborns that did not pass in 3 months. RESULT: In these 13 315 newborns, 5 151 subjects who did not pass the initial screening, 1910 subjects who also did not pass after 42 days, 1167 subjects cannot pass the rescreening after 3 months, 642 subjects were diagnosed congenital hearing impairment by Brainstem Auditory Evoked Potential Test, the rate is 4.82%. CONCLUSION: TEOAE and AABR are the suitable model of universal newborns' hearing screening in high-risk neonates.

Effect of ASCs on the graft survival rates of fat particles in rabbits.

This study was conducted to explore the effect of adipose-derived stem cells (ASCs) on the graft survival rates of fat particles in rabbits. Six domestic rabbits were used for a 3-month study; 1.4 grams of fat tissues were harvested from the bilateral inguinal regions of each animal. They were cut into granules and divided into three parts (A = 0.4 g, B = 0.4 g, and C = 0.6 g). Part A was centrifuged after 0.075% collagenase digestion for isolation of the stromal vascular fraction (SVF). About 0.2 grams of SVF containing ASCs was obtained, and then incorporated with part B to create a treated group, whereas part C was treated as a control group. The tissues in both groups were randomly transplanted into a subcutaneous space that had been created on each side of the dorsal midline of the rabbit. The grafts were taken out after 3 months for calculation of the survival rates. The graft survival rate in the treated group was 23.56 ± 2.49%, while that in the control group was 11.06 ± 2.10%. The graft survival rate in the treated group increased significantly, compared with in the control group (p < 0.01). Improved transplantation effects may be obtained by implanting the fat particles mixed with ASCs. It is suggested that this approach has the potential for becoming a new method of fat graft in clinical practice.

[Constructing duplication hasABC of chromosome recombinant in Streptococcus equi subsp. Zooepidemicu].

OBJECTIVE: To investigate the possibility of increasing the yield of hyaluronic acid by constructing duplication hasABC of chromosome recombinant in Streptococcus equi subsp. Zooepidemicus with a thermosensitive delivery vector system pJR700. METHODS: We amplified a 4147 bp DNA fragment of hyaluronic acid synthase operon hasABC genes from chromatosome of S. zooepidemicus using PCR. This DNA fragment was subcloned into the pJR700 at ClaI sites to result in recombinant plasmid pXL32. The recombinant plasmid was transformed into S. Zooepidemicus by electroperation. The homologous recombination was induced by growing the bacteria at 37 degrees C, and transformants were selected according to kanamycin resistance for 3 rounds. Then the culture was shifted to grow at 30 degrees C without antibiotics for 4 rounds to induce excision of the pJR700 indicated fragment. Colonies with kanamycin sensitivity were selected by plating on THY agar at 37 degrees C. The hasABC recombinant of S. Zooepidemicus was identified through RT-PCR with primers homologous to the flanking regions. HA titers were measured by the modified carbazole assay. RESULTS: We constructed successfully the duplication hasABC of chromosome recombinant of S. Zooepidemicus and the HA titer production by recombinants harboring duplication hasABC was 34% higher than that of the wild type at 24 h in shake flask culture. CONCLUSION: The thermosensitive delivery vector of pJR700 could be used to construct the streptococcal hasABC recombinant strain for increasing the yield of HA in S. Zooepidemicus.

[Construction of in-frame deletion streptococcal hemoprotein receptor gene mutant in Streptococcus equi subsp. zooepidemicus].

OBJECTIVE: To construct the in-frame deletion gene mutants in streptococcus equi subsp. zooepidemicusby (S. zooepidemicus) by pJR700 plasmid, a thermosensitive delivery vector system. METHOD: With PCR we amplified a 4017-bp DNA fragment of streptococcal hemoprotein receptor gene from chromatosome DNA. This DNA fragment was subcloned into the pJR700 plasmid to create pXL28. Using pXL28 as the template we got the DNA fragment deleted 1831 bp in streptococcal hemoprotein receptor gene by inverse PCR amplification and ligated the in-frame deletion DNA fragment by T4DNase to create pXL30. Then pXL30 was transformed into S. Zooepidemicus by electroperation. The kanamycin( kan)-resistant clones were selected at 37 degrees C in the presence of kan for three rounds. To induce excision of the plasmid vector sequence, the culture was shifted to 30 degrees C and grown without antibiotics for four rounds. Colonies with kan sensitivity, which indicated that the plasmid had been excised, were selected by plating on THY agar at 37 degrees C. S. Zooepidemicus mutants were identified through PCR with primers homologous to the flanking regions and the streptonigrin sensitive test. RESULT: The in-frame deletion streptococcal hemoprotein receptor mutants were successfully built in S. Zooepidemicus. CONCLUSION: The thermosensitive delivery vector system of pJR700 could be utilized to construct the in-frame deletion gene mutant strains of S. Zooepidemicus.