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Abstract [Objective] The woody stems of plants are difficult to obtain satisfactory microscopic image in both freehand and paraffin sections due to the high content of secondary vascular tissues and high degrees of lignification of cell walls. Microtome cryostat can section the tissue with a certain hardness. It will be one of the effective tools for the section of plant woody stems. [Methods] Some woody stems with uniform texture and high water content can be sectioned by cryo-sectioning without chemical fixation. For stems with hard wood and less water content, observation was performed after fixation with FAA and other fixation, OCT embedding, (-30) - (-25) ℃ sectioning, and staining with 0.1% toluidine blue staining solution. [Results] For tree or shrub stems with a diameter of less than 6 cm, cryo-sectioning method could be used to quickly obtain complete structure and clear and colorful microstructure images. [Conclusion] The microtome cryostat can be used for microstructure sectioning of woody stems combined with toluidine blue staining, which has the advantage of simplicity, efficiency, and ease of operation.

Objective: To better control the growth of spoilage microorganisms, and ensure the food safety of Xiangwei Luodou dry. Methods: The putrefactive microorganisms in Xiangwei dried tofu were identified by plate scribing method, colony morphology observation, Gram staining, 16S rDNA and ITS sequence. Results: A total of 9 strains were isolated andscreened. KZ2780-3 and KZ2780-4 are Bacillus subtilis, NZ2559-1 is Bacillus cereus, and JZ2559-5 is Bacillusamylolyticus amyloliquefaciens, FZ2559-2 for Klebsiella pneumoniae, SZ2641-5 for Enterococcus faecium, RZ2641-7 for Staphylococcus haemolyticus, BZ2780-1 for Acinetobacter baui, CZ2780-5 for Penicillium citri. Conclusion: The strains causing dry rot of Xiangwei bean are Bacillus subtilis, Bacillus cereus, Bacillus amyloliquefaciens, Klebsiella pneumoniae, Enterococcus faecium, Staphylococcus hemolyticus, Acinetobacter baui and Penicillium citri. The control methods of Xiangwei stewed bean dry are ultra-high temperature instantaneous sterilization technology, plasma sterilization technology, natural antibacterial substances combined with biological preservatives, vacuum packaging, air conditioning preservation and so on.

Cadmium (Cd) contamination in rice has been a serious threat to human health. To investigate the effects of arbuscular mycorrhizal fungi (AMF) on the Cd translocation in rice, a controlled pot experiment was conducted. The results indicated that AMF significantly increased rice biomass, with an increase of up to 40.0%, particularly in root biomass by up to 68.4%. Notably, the number of prominent rice individuals also increased, and their plasticity was enhanced following AMF inoculation. AMF led to an increase in the net photosynthetic rate and antioxidant enzyme activity of rice. In the AMF treatment group, the Cd concentration in the rice roots was significantly higher (19.1%‒68.0%) compared with that in the control group. Conversely, the Cd concentration in the rice seeds was lower in the AMF treatment group, indicating that AMF facilitated the sequestration of Cd in rice roots and reduced Cd accumulation in the seeds. Path coefficients varied across different treatments, suggesting that AMF inoculation reduced the direct impact of soil Cd concentration on the total Cd accumulation in seeds. The translocation of Cd was consistently associated with simultaneous growth dilution and compensatory accumulation as a result of mycorrhizal effects. Our study quantitatively analyzed this process through path analysis and clarified the causal relationship between rice growth and Cd transfer under the influence of AMF.

Abstract [Objective] This study aims to investigate the impact of Streptomyces pactum Act12 broth on growth and physiological characteristics of Medicago sativa seedlings under petroleum stress. [Methods] The effects of different concentrations (0, 1%, 10%, and 100%) of Act12 broth under a petroleum stress level of 3% (W/W ) were examined by determining seedling growth, root morphology, photosynthetic characteristics, osmoregulatory substance content, and antioxidant capacity in M . sativa. [Results] The 3% petroleum significantly hindered photosynthesis and biomass accumulation of M . sativa seedlings, compared to the control group (CK). Moreover, it resulted in elevation in malondialdehyde level, decrease in soluble sugar and soluble protein content, as well as alteration in the activities of superoxide dismutase (SOD) and peroxidase (POD). Compared to the absence of Act12 broth, solutions with low and medium concentrations (1% and 10%) effectively mitigated the toxic effects of petroleum, with 10% Act12 broth exhibiting the most significant effect. The seedlings treated with Act12 broth exhibited significant enhancements in plant height, above-ground dry mass, underground dry mass, stem tolerance index, root tolerance index, total root length, root surface area, root volume, and number of root tips. Additionally, the level of MDA was decreased by 42.49% and 56.45%, soluble sugar was increased by 79.25% and 89.83%, while soluble protein was increased by 167.63% and 256.15% in the above-ground and underground parts, respectively. Moreover, SOD and POD activity were increased by 35.81% and 57.33%, respectively. [Conclusion] The fermented liquid with low/medium concentration effectively modulates antioxidant enzyme activity and osmotic regulation ability of seedlings, resulting in reduced MDA content, enhanced photosynthesis ability, increased biomass, and eventually alleviates the inhibition effects of petroleum stress on seedling growth.

The hot-rolled Al-Zn-Mg-Cu system aluminum alloy was treated with single-stage solution，low temperature-long term & high temperature-short term and low temperature-short term & high temperature-long term，double-stage solution quenching systems. The effects of different solution methods on the structure and properties of Al-Zn-Mg-Cu system aluminum alloy were systematically compared and analyzed by OM，SEM，DSC，EBSD and other technical means，as well as conductivity testing methods. The results show that the selected solution quenching system will not cause the structure to overheat，and can fully dissolve the eutectic second phase of the low melting point in the structure. When 473 ℃/20 min+477 ℃/40 min two-stage solution quenching system is selected，the plate has the best solution effect and the lowest conductivity，which is 27.60%IACS，and the recrystallization degree of the structure is less than 50%. There are {112}〈111〉，{011}〈211〉，{123}〈634〉 deformation texture and {001}〈110〉 shear texture in the structure.

Hypertrophic scar (HS) is one of the most common sequelae of patients, especially after burns and trauma. The roles of regulatory long noncoding RNAs (lncRNAs) in mediating HS remain underexplored. Human hypertrophic scar-derived fibroblasts (HSFBs) have been shown to exert more potent promoting effects on extracellular matrix (ECM) accumulation than normal skin-derived fibroblasts (NSFBs) and are associated with enhanced HS formation. The purpose of this study is to search for lncRNAs enriched in HSFBs and investigate their roles and mechanisms. LncRNA MSTRG.59347.16 is one of the most highly expressed lncRNAs in HS detected by lncRNA-seq and qRT-PCR and named as hypertrophic scar fibroblast-associated lncRNA (HSFAS). HSFAS overexpression significantly induces fibroblast proliferation, migration, and myofibroblast trans-differentiation and inhibits apoptosis in HSFBs, while knockdown of HSFAS results in augmented apoptosis and attenuated proliferation, migration, and myofibroblast trans-differentiation of HSFBs. Mechanistically, HSFAS suppresses the expression of A disintegrin and metalloproteinase with thrombospondin motifs 8 (ADAMTS8). ADAMTS8 knockdown rescues downregulated HSFAS-mediated fibroblast proliferation, migration, myofibroblast trans-differentiation and apoptosis. Thus, our findings uncover a previously unknown lncRNA-dependent regulatory pathway for fibroblast function. Targeted intervention in the HSFAS-ADAMTS8 pathway is a potential therapy for HS.

Long noncoding RNAs (lncRNAs) are increasingly being implicated as key regulators of cell proliferation, apoptosis, and differentiation. However, the molecular mechanisms of specific lncRNAs in the context of hypertrophic scar remain largely unclear. Here, we find that the lncRNA FPASL (fibroblast proliferation-associated LncRNA) is downregulated in HS, and FPASL reduces fibroblast proliferation and colony formation and blocks cell cycle progression. Using GO annotation enrichment analysis along with AZC (a specific inhibitor of DNA methylation), we identify that DNA methylation is responsible for downregulating FPASL in hypertrophic scar. Subsequent studies demonstrate that high expression of DNMT3b inhibits FPASL expression in HS. Mechanistic study reveals a significant increase in fibroblast proliferation after transfection with LNA-FPASL, which is further inhibited by knockdown of DNMT3b. Thus, our study reveals that DNMT3b mediates hypermethylation of the lncRNA FPASL promoter and the downregulation of lncRNA FPASL promotes fibroblast proliferation in hypertrophic scar.

Immune-related cardiotoxicities are uncommon but potentially fatal. The study aims to evaluate the value of pacemakers and methylprednisolone pulse therapy (MPPT) to patients with immune-related myocarditis concomitant with complete heart block (CHB). We first reviewed medical records of three patients with immune-related myocarditis concomitant with CHB. For the pooled analysis, we searched related cases with immune-related myocarditis in the PubMed database and screened the patients. Clinical characteristics, management, and outcomes were summarized. Our three patients developed immune-related myocarditis concomitant with CHB about 2 weeks after receiving pembrolizumab, and were successfully treated with pacemaker implantation and high-dose steroids (two received MPPT). In the pooled analysis, 21 cases were eligible with an overall fatality rate of 52%. Patients with pacemakers had a fatality rate of 38%, significantly lower than patients without them (38% vs 100%; p = 0.035), particularly the MPPT subgroup (25% vs 100%; p = 0.019). All five patients without pacemakers expired. Among patients with pacemakers, MPPT patients tended to have an inferior rate compared with non-MPPT patients. Timely pacemaker implantation played a crucial role in improving the outcomes of patients with immune-related myocarditis concomitant with CHB. Patients receiving MPPT appeared to have a better prognosis. Additionally, multidisciplinary consultation should be recommended for better management.

Hypertrophic scar is a problem for numerous patients, especially after burns, and is characterized by increased fibroblast proliferation and collagen deposition. Increasing evidence demonstrates that lncRNAs contribute to the development and progression of various diseases. However, the function of lncRNAs in hypertrophic scar formation remains poorly characterized. In this study, a novel fibroblast proliferation-associated lncRNA, named lncRNA FPASL (MSTRG.389905.1), which is mainly localized in the cytoplasm, is found to be downregulated in hypertrophic scar, as detected by lncRNA microarray and qRT-PCR. The full-length FPASL is characterized and further investigation confirms that it has no protein-coding potential. FPASL knockdown in fibroblasts triggers fibroblast proliferation, whereas overexpression of FPASL directly attenuates the proliferation of fibroblasts. Furthermore, target genes of the differentially expressed lncRNAs in hypertrophic scars and the matched adjacent normal tissues are enriched in fibroblast proliferation signaling pathways, including the PI3K/AKT and MAPK signaling pathways, as determined by GO annotation and KEGG enrichment analysis. We also demonstrate that knockdown of FPASL activates the PI3K/AKT and MAPK signaling pathways, and specific inhibitors of the PI3K/AKT and MAPK signaling pathways can reverse the proliferation of fibroblasts promoted by FPASL knockdown. Our findings contribute to a better understanding of the role of lncRNAs in hypertrophic scar and suggest that FPASL may act as a potential novel therapeutic target for hypertrophic scar.

Objective To investigate the correlation between the expression of Hsa_circ_0026352 and the clinical characteristics of breast cancer(BC) patients, to evaluate the value of Hsa_circ_0026352 as a diagnostic marker of breast cancer. Methods Human circRNA microarray was used to screen the different expression of circRNAs in BC tissues. qRT-PCR was used to verify the expression of Hsa_circ_0026352 in BC tissue and peripheral blood. CircRNA structure were performed by circPrimer1.2 software. T-test, ANOVA analysis, curve regression analysis and ROC curve analysis were performed to determine the diagnostic values of Hsa_circ_0026352. Results Hsa_circ_0026352 was significantly down-regulated in both breast cancer tissues and peripheral blood (P < 0.01). The AUC of Hsa_circ_0026352 were 0.881 in tissues and 0.826 in peripheral blood (both P < 0.01). The relative expression of Hsa_circ_0026352 in peripheral blood of BC cancer patients was negatively correlated with CA153 level (P < 0.05). The AUC of Hsa_circ_0026352 in peripheral blood of patients with ER positive, early stage breast cancer and breast neoplasm metastasis were 0.710, 0.771 and 0.722 (all P < 0.01), respectively. Conclusion Hsa_circ_0026352 could be a novel predictive biomarker for diagnosis of BC.

This paper investigates a method to dynamically model compound faults on the inner and outer rings of an angular contact ball bearing as well as their effects on its dynamic behavior. Gupta’s dynamic modeling method is used to consider changes in the deformation and direction of the contact load when the ball passes through the damaged area and to develop a dynamic model of compound faults in the angular contact ball bearing. The step-changing fourth-order Runge–Kutta method is used to solve the dynamic compound fault model. The time-domain signal of vibration responses in the case of a single fault in the inner and outer rings exhibited a certain periodicity, and the frequency of faults in the envelope spectrum was clear. By comparison, the periodicity of compound faults was not clear. The signals of compound faults were decomposed by the dual-tree complex wavelet transform to identify their characteristic frequency. Errors occurred between the characteristic frequency of the theoretical fault and its simulated value. They increased with the rotational speed and decreased with an increase in axial load, whereas the influence of radial load on them was minor. For compound faults on the inner and outer rings of an angular contact ball bearing, this study provides a modeling method that can describe changes in the deformation and direction of the contact load when the ball passes through the damaged area of the inner and outer rings. The work here can provide an important foundation for fault identification in angular contact ball bearings.

C15H16N2O3, tetragonal, I41/a (no. 88), a = 14.646(2) Å, c = 26.028(2) Å, V = 5583.4 Å3, Z = 16, Rgt(F) = 0.0497, wRref(F2) = 0.1680, T = 298 K.

Abstract Alcohol-induced injury has become one of the major causes for liver cirrhosis. However, the molecular mechanisms of ethanol-induced injury are not fully understood. To this end, we performed a dynamic plasma membrane proteomic research on rat model. A rat model from hepatitis to liver cirrhosis was developed. Plasma membrane from liver tissue with liver fibrosis stage of 2 and 4 (S2 and S4) was purified by sucrose density gradient centrifugation. Its purification was verified by western blotting. Proteins from plasma membrane were separated by two-dimensional electrophoresis (2DE) and differentially expressed proteins were identified by tandem mass spectrometry. 16 consistent differentially expressed proteins from S2 to S4 were identified by mass spectrometry. The expression of differentially expressed proteins annexin A6 and annexin A3 were verified by western blotting, and annexin A3 was futher verified by immunohistochemistry. Our research suggests a possible mechanism by which ethanol alters protein expression to enhance the liver fibrosis progression. These differentially expressed proteins might be new drug targets for treating alcoholic liver cirrhosis.

Abstract Background Osteosarcoma (OS) is the most common primary malignant tumor of bone in children and adolescents. However, the knowledge in diagnostic modalities has progressed less. To identify new biomarkers for the early diagnosis of OS as well as for potential novel therapeutic candidates, we performed a sub-cellular comparative proteomic research. Methods An osteosarcoma cell line (MG-63) and human osteoblastic cells (hFOB1.19) were used as our comparative model. Plasma membrane (PM) was obtained by aqueous two-phase partition. Proteins were analyzed through iTRAQ-based quantitative differential LC/MS/MS. The location and function of differential proteins were analyzed through GO database. Protein-protein interaction was examined through String software. One of differentially expressed proteins was verified by immunohistochemistry. Results 342 non-redundant proteins were identified, 68 of which were differentially expressed with 1.5-fold difference, with 25 up-regulated and 43 down-regulated. Among those differential proteins, 69% ware plasma membrane, which are related to the biological processes of binding, cell structure, signal transduction, cell adhesion, etc., and interaction with each other. One protein--CD151 located in net nodes was verified to be over-expressed in osteosarcoma tissue by immunohistochemistry. Conclusion It is the first time to use plasma membrane proteomics for studying the OS membrane proteins according to our knowledge. We generated preliminary but comprehensive data about membrane protein of osteosarcoma. Among these, CD151 was further validated in patient samples, and this small molecule membrane might be a new target for OS research. The plasma membrane proteins identified in this study may provide new insight into osteosarcoma biology and potential diagnostic and therapeutic biomarkers.

Abstract Background The human immunodeficiency virus type 1 (HIV-1) pandemic has continued unabated for nearly 30 years. To better understand the influence of virus on host cells, we performed the differential proteome research of peripheral blood mononuclear cells (PBMCs) from HIV-positive patients and healthy controls. Results 26 protein spots with more than 1.5-fold difference were detected in two dimensional electrophoresis (2DE) gels. 12 unique up-regulated and one down-regulated proteins were identified in HIV-positive patients compared with healthy donors. The mRNA expression of 10 genes was analyzed by real time RT-PCR. It shows that the mRNA expression of talin-1, vinculin and coronin-1C were up-regulated in HIV positive patients and consistent with protein expression. Western blotting analysis confirmed the induction of fragments of vinculin, talin-1 and filamin-A in pooled and most part of individual HIV-positive clinical samples. Bioinformatic analysis showed that a wide host protein network was disrupted in HIV-positive patients. Conclusions Together, this work provided useful information to facilitate further investigation of the underlying mechanism of HIV and host cell protein interactions, and discovered novel potential biomarkers such as fragment of vinculin, filamin-A and talin-1 for anti-HIV research.