Recent studies have indicated that the expression of ubiquitin-specific protease 51 (USP51), a novel deubiquitinating enzyme (DUB) that mediates protein degradation as part of the ubiquitin‒proteasome system (UPS), is associated with tumor progression and therapeutic resistance in multiple malignancies. However, the underlying mechanisms and signaling networks involved in USP51-mediated regulation of malignant phenotypes remain largely unknown. The present study provides evidence of USP51's functions as the prominent DUB in chemoresistant triple-negative breast cancer (TNBC) cells. At the molecular level, ectopic expression of USP51 stabilized the 78 kDa Glucose-Regulated Protein (GRP78) protein through deubiquitination, thereby increasing its expression and localization on the cell surface. Furthermore, the upregulation of cell surface GRP78 increased the activity of ATP binding cassette subfamily B member 1 (ABCB1), the main efflux pump of doxorubicin (DOX), ultimately decreasing its accumulation in TNBC cells and promoting the development of drug resistance both in vitro and in vivo. Clinically, we found significant correlations among USP51, GRP78, and ABCB1 expression in TNBC patients with chemoresistance. Elevated USP51, GRP78, and ABCB1 levels were also strongly associated with a poor patient prognosis. Importantly, we revealed an alternative intervention for specific pharmacological targeting of USP51 for TNBC cell chemosensitization. In conclusion, these findings collectively indicate that the USP51/GRP78/ABCB1 network is a key contributor to the malignant progression and chemotherapeutic resistance of TNBC cells, underscoring the pivotal role of USP51 as a novel therapeutic target for cancer management.

Objective:To prepare a humanized monoclonal antibody against periostin and establish a stable cell line.Meth-ods:Based on anti-periostin mouse monoclonal antibody developed by our laboratory,total RNA was extracted,and variable region sequences were obtained by RT-PCR amplification of VH and VL genes.The mouse antibody CDR region was transplanted into the human antibody framework receptor region,and the gene was subcloned into the expression vector PATX-GS2,and stably transfected into CHO cells.Monoclonal cell lines were obtained by MSX pressure screening and limited dilution.Results:VH and VL genes were amplified by RT-PCR,and the sequence of the light and heavy chain variable region were determined.Antibody humanization were successfully stablished by CDR transplantation method a murine antibody to a human framework,and a eukaryotic expression plasmid was constructed,which was transfected into CHO cells for expression,and human anti-periostin antibody was successfully obtained.ELISA and Western blot results showed that the humanized antibody had good anti-periostin activities and binding affinity.Conclu-sion:In this study,anti-periostin humanized monoclonal antibody has been successfully prepared,which can specifically bind to peri-ostin proteins in vivo and have biological activity,providing scientific data for the precise treatment of retinal fibrosis,tissue and organ fibrosis,and malignant tumors.

This study aims to explore the anti-depression mechanism of Zuojin Pills based on the plasma constituents, network pharmacology, and experimental verification. UHPLC-TOF-MS was used for qualitative analysis of Zuojin Pills-containing serum. Targets of the plasma constituents and the disease were retrieved from PharmMapper and GeneCards. Then the protein-protein interaction(PPI) network was constructed and core targets were screened for GO term enrichment and KEGG pathway enrichment. Cytoscape 3.7.2 was employed construct the "compound-target-pathway" network and the targets and signaling pathways of Zuojin Pills against depression were predicted. CUMS-induced depression mouse model was established to verify the key targets. The results showed that a total of 21 constituents migrating to blood of Zuojin Pills were identified, which were mainly alkaloids. A total of 155 common targets of the constituents and the disease and 67 core targets were screened out. KEGG enrichment and PPI network analysis showed that Zuojin Pills may play a role in the treatment of depression through AMPK/SIRT1, NLRP3, insulin and other targets and pathways. Furthermore, the results of animal experiments showed that Zuojin Pills could significantly improve the depression behaviors of depression, reduce the levels of IL-1β, IL-6 and TNF-α in hippocampus and serum, activate AMPK/SIRT1 signaling, and reduce the protein expression of NLRP3. In conclusion, Zuojin Pills may play a role in the treatment of depression by activating AMPK/SIRT1 signaling pathway, and inhibiting NLRP3 activation and neuroinflammation in the hippocampus of mice.
Animals ; Mice ; Network Pharmacology ; AMP-Activated Protein Kinases ; Chromatography, High Pressure Liquid ; NLR Family, Pyrin Domain-Containing 3 Protein ; Sirtuin 1 ; Drugs, Chinese Herbal/pharmacology* ; Molecular Docking Simulation

Aim To preliminarily investigate the effect of brusatol(BRU), the monomer components fromChinese medicines on H1299 cells and its mechanisms.Methods CCK-8 and EdU staining experiment were used to detect the effect of BRU on cell prolifera-tion.Clone formation experiment was performed to measure the effect of drugs on cell clone formation.Hoechst33258 staining experiment and flow cytometry were employed to observe the cell apoptosis.Western blot assay was used to detect the protein expression levels of Bcl-xL, Bax, Bcl-2, cleaved-caspase-3, caspase-3, Gadd45α, PI3K, p-PI3K, Akt, p-Akt and NF-κB-p65.Results CCK-8 assay revealed that the inhibitory effect of H1299 cells gradually increased with the rising of BRU concentration and action time.Compared with control group, the EdU incorporation rate of the BRU treatment group decreased significantly.Treated with different concentrations of BRU for 24 h, the clone formation rate was significantly reduced in a concentration-dependent manner.Hoechst33258 experiment and flow cytometry showed that BRU could induce apoptosis in H1299 cell nucleus and increase its apoptotic rate.Western blot results revealed that BRU could significantly up-regulate the protein levels of Bax, cleaved-caspase-3, Gadd45α, and significantly down-regulate the expression of Bcl-xL, Bcl-2, caspase-3.In addition, BRU could significantly decrease the expression level of p-PI3K, p-Akt, NF-κB-p65 in cell nucleus.Conclusions BRU can inhibit the proliferation and induce apoptosis of H1299 cells in a concentration and time-dependent manner.The mechanism may be related to the inhibition of PI3K/Akt signaling pathway and the nuclear shuttle of NF-κB-p65.

Biochemistry is a very important basic course for biology and medical disciplines, and it is one of the most rapidly developing and dynamic disciplines since the 21st century. Since theoretical teaching of biochemistry is very abstract, experiment teaching is an important part of understanding relevant theories and mastering practical skills. In the process of experiment teaching, it is always a key concern for teachers to keep abreast of new teaching concepts, new teaching methods, new teaching hotspots, and keep up with teaching trends. To this end, in this paper, the annual number of published papers, periodical distribution, research keywords, current status of research, hotspots and development trends of biochemical experiment teaching and research in China from 2011. 1. 1 to 2020. 12. 31 were visualized and analyzed by using CiteSpace with China National Knowledge Infrastructure (CNKI) as the data source. It was found that the number of articles published in each year remained basically stable, and the high frequency words were " biochemistry", " experimental teaching", " teaching reform", " biochemistry experiment", " reform", " teaching method", " innovation ability" and " teaching mode". The words with higher intensity are " flipped classroom" and " microlearning". Through timeline analysis, we found that the identities of " teaching" and " learning" are being exchanged, and improving students' independent learning ability has become the mainstream. The teaching mode combining advanced technology such as " virtual simulation" and " online teaching" has become the development trend and frontier of biochemistry experiment teaching. This paper aims to provide reference for the sustainable development of biochemistry experiment teaching in China through the above analysis.

Objective: To explore the protective effects of anthrahydroquinone-2,6-disulfonate (AH 2 QDS) on the kidneys of paraquat (PQ) poisoned rats via the apelin-APJ pathway. Methods: Male Sprague Dawley rats were divided into four experimental groups: control, PQ, PQ+sivelestat, and PQ+AH 2 QDS. The PQ+sivelestat group served as the positive control group. The model of poisoning was established via intragastric treatment with a 20% PQ pesticide solution at 200 mg/kg. Two hours after poisoning, the PQ+sivelestat group was treated with sivelestat, while the PQ+AH 2 QDS group was given AH 2 QDS. Six rats were selected from each group on the first, third, and seventh days after poisoning and dissected after anesthesia. The PQ content of the kidneys was measured using the sodium disulfite method. Hematoxylin-eosin staining of renal tissues was performed to detect pathological changes. Apelin expression in the renal tissues was detected using immunofluorescence. Western blotting was used to detect the expression levels of the following proteins in the kidney tissues: IL-6, TNF-α, apelin-APJ (the apelin-Angiotensin receptor), NF-κB p65, caspase-1, caspase-8, glucose-regulated protein 78 (GRP78), and the C/EBP homologous protein (CHOP). In in vitro study, a PQ toxicity model was established using human tubular epithelial cells treated with standard PQ. Twenty-four hours after poisoning, sivelestat and AH 2 QDS were administered. The levels of oxidative stress in human renal tubular epithelial cells were assessed using a reactive oxygen species fluorescence probe. Results: The PQ content in the kidney tissues of the PQ group was higher than that of the PQ+AH 2 QDS group. Hematoxylin-eosin staining showed extensive hemorrhage and congestion in the renal parenchyma of the PQ group. Vacuolar degeneration of the renal tubule epithelial cells, deposition of crescent-like red staining material in renal follicles, infiltration by a few inflammatory cells, and a small number of cast formation were also observed. However, these pathological changes were less severe in the PQ+sivelestat group and the PQ+AH 2 QDS group (P<0.05). On the third day after poisoning, immunofluorescence assay showed that the level of apelin in the renal tissues was significantly higher in the PQ+AH 2 QDS group than in the PQ group. Western blotting analysis results showed that IL-6, TNF-α, NF-κB p65, caspase-1, caspase-8, GRP78, and CHOP protein levels in the PQ group were higher than in the PQ+AH 2 QDS group (P<0.05). The expression of apelin-APJ proteins in the PQ+AH 2 QDS group was higher than in the PQ+sivelestat and PQ groups (P<0.05); this difference was significant on Day 3 and Day 7. The level of oxidative stress in the renal tubular epithelial cells of the PQ+AH 2 QDS group and the PQ+sivelestat group was significantly lower than in the PQ group (P<0.05). Conclusions: This study confirms that AH 2 QDS has a protective effect on PQ-poisoned kidneys and its positive effect is superior to that of sivelestat. The mechanism of the protective effects of AH 2 QDS may be linked to reduction in cellular oxidative stress, PQ content of renal tissue, inflammatory injury, endoplasmic reticulum stress, and apoptosis. AH 2 QDS may play a role in the treatment of PQ poisoning by upregulating the expression of the apelin-APJ.

Objective: To investigate the clinical efficacy of long-segment pedicle screw reduction and internal fixation combined with kyphoplasty in the treatment of stage Ⅲ reducible Kummell disease. Methods: The clinical data of 32 patients with stage Ⅲ reducible Kummell disease treated at the Department of Orthopedics, Sir Run Run Shaw Hospital, Zhejiang University College of Medicine from January 2012 to March 2017 were analyzed retrospectively.There were 7 males and 25 females,aged (71.8±6.7)years(range:61 to 86 years).The injured segment was T₁₀ in 1 patient,T₁₁ in 8 patients,T₁₂ in 13 patients,L₁ in 7 patients,L₂ in 2 patients and L₃ in 1 patient.Preoperative American spinal injury association(ASIA) classification of patients all showed grade D.Bone mineral density (BMD),spinal X-ray,CT and MRI were examined before operation.All patients were treated with postural reduction, long-segment pedicle screw reduction and internal fixation combined with kyphoplasty.The operation time,intraoperative blood loss,length of stay and postoperative complications were recorded.The visual analogue scale (VAS) and Oswestry dysfunction index (ODI) as well as the BMD of hip were collected before and after operation.The Cobb angle of involved segment kyphosis and the height of anterior edge of diseased vertebrae were measured before operation,3 days and 12 months after operation.CT-related parameters were measured before and 3 days after operation,including sagittal anterior and posterior diameter of spinal canal,cross-sectional anterior and posterior diameter of spinal canal and cross-sectional spinal canal area.Paired sample t test and repeated measures were used to compare the data before and after operation. Results: All patients received the operation successfully.The operation time was (131.3±16.9) minutes (range:95 to 180 minutes),the blood loss was (82.5±27.1) ml (range:50 to 150 ml),and the length of stay was (8.3±2.4) days (range:5 to 14 days).All patients were followed up for more than 12 months.The VAS decreased gradually at 3 days,3 months,6 months and 12 months after operation,and the differences were statistically significant compared with the VAS before surgery (all P<0.01).ODI at 3,6 and 12 months after surgery was significantly improved compared with that before surgery(All P<0.01).The CT-related parameters at 3 days after operation were significantly higher than those before operation (All P<0.05).At 12 months after surgery,the Cobb angle decreased from (35.2±7.6) ° preoperatively to (4.3±1.7) ° (t=22.630,P<0.01),the height of anterior edge of diseased vertebrae increased from (4.3±1.0) mm preoperatively to (16.9±2.5) mm(t=-25.845,P<0.01),the bone mineral density of hip increased from -(2.2±0.6) preoperatively to -(2.8±0.6)(t=-0.040,P<0.01).Up to the last follow-up,2 patients had distal pedicle screw loosening, 1 patient had proximal junctional kyphosis,and there was no new vertebral fracture. Conclusions: Based on postural reduction,long-segment pedicle screw reduction and internal fixation combined with kyphoplasty is a safe and effective treatment method for stage Ⅲ reducible Kummell disease,which can reconstruct the stability of the diseased vertebrae.Postoperative standard anti-osteoporosis treatment is the basis to ensure the efficacy.
Aged ; Cross-Sectional Studies ; Female ; Fracture Fixation, Internal ; Humans ; Kyphoplasty ; Lumbar Vertebrae/surgery* ; Male ; Pedicle Screws ; Retrospective Studies ; Spinal Fractures/surgery* ; Thoracic Vertebrae/surgery* ; Treatment Outcome

Aim To explore the effect of homoharringtonine (HHT) on the prohferation of liver cancer cell PLCS and its possible mechanism. Methods CCK-8 and EdU were used to detect the effect of HHT on the proliferation of PLCS cells; flow cytometry was employed to assess the effect of HHT on cell cycle of PLCS; Western blot was applied to measure the expression levels of cycle-related proteins cyclinA, CDK 2, p 2 1, p53 and A T M. Results Treated with HHT (0, 5, 10, 20, 40, 80 • L