SRSF3 suppresses RCC tumorigenesis and progression via regulating SP4 alternative splicing.

Abnormal alternative splicing (AS) caused by dysregulated expression of splicing factors plays a crucial role in tumorigenesis and progression. The serine/arginine-rich (SR) RNA-binding protein family is a major class of splicing factors regulating AS. However, their roles and mechanisms in renal cell carcinoma (RCC) development and progression are not fully understood. Here, we found that SR splicing factor 3 (SRSF3) was an important splicing factor affecting RCC progression. SRSF3 was downregulated in RCC tissues and its low level was associated with decreased overall survival time of RCC patients. SRSF3 overexpression suppressed RCC cell malignancy. Mechanistically, the binding of SRSF3 to SP4 exon 3 led to the inclusion of SP4 exon 3 and the increase of long SP4 isoform (L-SP4) level in RCC cells. L-SP4, but not S-SP4 overexpression suppressed RCC cell malignancy. Meanwhile, L-SP4 participated in SRSF3-mediated anti-proliferation by transcriptionally promoting SMAD4 expression. Taken together, our findings provide new insights into the anticancer mechanism of SRSF3, suggesting that SRSF3 may serve as a novel potential therapeutic target for RCC.

Advancing treatment strategies for posterior malleolar malunion: The ankle dislocation method.

BACKGROUND: This study aimed to assess the radiological and clinical outcomes of treatment using the ankle dislocation method for posterior malleolar malunion. METHOD: Thirty-one patients with posterior malleolar malunion who underwent treatment using the ankle dislocation method from May 2015 to October 2021 were retrospectively analyzed. Key outcome measures were radiographic parameters (articular step-off, tibiofibular clear space, fibular length, tibial lateral surface angle, and ankle osteoarthritis), clinical scores (American Orthopaedic Foot and Ankle Society ankle-hindfoot scale and Visual Analogue Scale), and patient satisfaction rate. RESULT: Preoperative computed tomography revealed that Bartoní cek types 3 and 4 accounted for 64.5 % (n = 20) of total cases. Most posterior malleolar malunions were accompanied by depressed intercalary fragments (61.2 % [n = 19]). At the final follow-up, radiographic parameters and clinical scores showed significant improvements postoperatively (P < 0.05), with a high patient satisfaction rate of 77.4 %. Subgroup analysis revealed that the posterior malleolar fracture morphology significantly affected postoperative pain, particularly in more complex fractures (P < 0.001). CONCLUSION: The ankle dislocation method effectively exposes the distal tibial articular surface and facilitates the anatomical restoration of joint congruity under direct vision. This approach substantially improves the clinical and imaging outcomes in patients with complex posterior malleolar malunion. LEVELS OF EVIDENCE: Level IV, retrospective case series.

PDZK1 suppresses TNBC development and sensitizes TNBC cells to erlotinib via the EGFR pathway.

Epidermal growth factor receptor (EGFR)-targeted drugs (erlotinib, etc.) are used to treat multiple types of tumours. EGFR is highly expressed in most triple-negative breast cancer (TNBC) patients. However, only a small proportion of TNBC patients benefit from EGFR-targeted drugs in clinical trials, and the resistance mechanism is unclear. Here, we found that PDZ domain containing 1 (PDZK1) is downregulated in erlotinib-resistant TNBC cells, suggesting that PDZK1 downregulation is related to erlotinib resistance in TNBC. PDZK1 binds to EGFR. Through this interaction, PDZK1 promotes EGFR degradation by enhancing the binding of EGFR to c-Cbl and inhibits EGFR phosphorylation by hindering EGFR dimerisation. We also found that PDZK1 is specifically downregulated in TNBC tissues and correlated with a poor prognosis in TNBC patients. In vitro and in vivo functional assays showed that PDZK1 suppressed TNBC development. Restoration of EGFR expression or kinase inhibitor treatment reversed the degree of cell malignancy induced by PDZK1 overexpression or knockdown, respectively. PDZK1 overexpression sensitised TNBC cells to erlotinib both in vitro and in vivo. In conclusion, PDZK1 is a significant prognostic factor for TNBC and a potential molecular therapeutic target for reversing erlotinib resistance in TNBC cells.

Changes in clinical outcomes and alignment of the ipsilateral knee and ankle after supramalleolar osteotomy in patients with varus osteoarthritis of the ankle: a short-term follow-up study.

PURPOSE: The purpose of this study was to investigate the changes in clinical outcomes and alignment of the ipsilateral knee and ankle in patients with varus ankle osteoarthritis after supramalleolar osteotomy (SMO). METHODS: We retrospectively reviewed 23 patients (24 ankles) with Takakura II, IIIa and IIIb ankle osteoarthritis treated with SMO between May 2017 and March 2022. The radiologic parameters of ankles contained medial distal tibial angle (TAS), tibiotalar angle (TT), tibial lateral surface (TLS), tibial plafond inclination (TPI) and talar inclination (TI). The radiologic parameters of knees contained medial proximal tibial angle (MPTA), joint line convergence angle (JLCA), the knee joint line orientation relative to ground (G-KJLO) and WBL. Hip-knee-ankle angle (HKA) was also collected. The Takakura system was used for evaluating the ankle osteoarthritis and the Kellgren-Lawrence (KL) system was used for evaluating the knee osteoarthritis. Clinical evaluation of the ankle joints contained American Orthopedic Foot and Ankle Society (AOFAS), range of motion (ROM) and visual analogue scale (VAS). Clinical evaluation of the knee joints contained Japanese Orthopaedic Association Scores (JOA), ROM, VAS. RESULTS: The mean follow-up times were 20.3 ± 7.3 months (range 12-38). According to the radiologic evaluation, the TAS increased from preoperative 84.7° ± 2.0° to 91.2° ± 1.8° at the last follow-up (P < 0.001). The TPI and TI decreased from 4.4° ± 4.2° and 11.0° ± 5.2° to 0.1° ± 4.7° and 4.1° ± 4.8° (P < 0.001 for both). The TT angel improved from 9.5° ± 4.1° to 4.9° ± 3.3° (P < 0.001). No significant differences were found regarding MPTA, JLCA, G-KJLO, knee WBL and HKA (P > 0.05 for all). The Takakura stage improved after SMO (P < 0.001) whilst the KL stage maintains the similar lever (P > 0.05). According to the clinical evaluation, the AOFAS significantly increased from 67.5 ± 10.6 to 88.5 ± 9.3 and the VAS of the ankle decreased from 4.7 ± 1.6 to 1.2 ± 1.1, whilst there were no changes on VAS and even the JOA and knee ROM after SMO (P > 0.05 for all). CONCLUSIONS: SMO can alleviate the symptoms of varus ankle osteoarthritis and delay the time for ankle replacement or arthrodesis by redistributing the abnormal stress of the ankle and restoring the congruence of the tibiotalar joint. In addition, it did not induce the clinical symptoms of knee without compromising lower limb alignment or knee joint line orientation in the short term. LEVEL OF EVIDENCE: Level IV case series.

Medial malleolar window approach for varus-type tibial pilon fractures: a retrospective study.

PURPOSE: Choosing a suitable surgical approach is crucial and challenging for type C pilon fractures. This article aims to explore the clinical efficacy of the medial malleolar window approach for varus-type tibial pilon fractures. METHODS: A retrospective analysis was conducted on 38 patients with type C varus-type pilon fractures treated between May 2018 and June 2021. In total, 16 cases underwent surgical treatment through the medial malleolar window approach and 22 cases were treated with the traditional anteromedial approach combined with a posterior approach. The operation time, hospitalization time, fracture healing time, the American Orthopedic Foot and Ankle score, Visual Analogue Scale, and complications were recorded to comprehensively evaluate the clinical efficacy of the technique. Fracture reduction quality was evaluated using the criteria proposed by Burwell and Charnley. RESULTS: All patients were followed up. No patients presented delayed union or nonunion. Compared with the conventional approach, the medial malleolar window approach had the advantage of better clinical effect recovery and better fracture reduction (P < 0.05). Meanwhile, the medial malleolar window approach had a shorter operation time, although the statistics suggest no significant difference with the control group. No implant exposure or infection occurred. There was good wound healing at two weeks after surgery in all but two cases. Local wound edge necrosis developed in one case in the medial malleolar window approach group, and the wound could not be closed at one stage in another case in the conventional group because of excessive tension, requiring secondary closure. CONCLUSION: The medial malleolar window approach provides excellent exposure to type C pilon fractures, allowing for satisfactory fracture reduction and functional rehabilitation. The medial window approach is recommended for varus-type pilon fractures, which can effectively avoid a posterior incision and reduce the operation time.

CRISPR/Cas9-Mediated Knockout of tnfaip1 in Zebrafish Plays a Role in Early Development.

TNF α-induced protein 1 (TNFAIP1) was first identified in human umbilical vein endothelial cells and can be induced by tumor necrosis factor α (TNFα). Early studies have found that TNFAIP1 is involved in the development of many tumors and is closely associated with the neurological disorder Alzheimer's disease. However, little is known about the expression pattern of TNFAIP1 under physiological conditions and its function during embryonic development. In this study, we used zebrafish as a model to illustrate the early developmental expression pattern of tnfaip1 and its role in early development. First, we examined the expression pattern of tnfaip1 during early zebrafish development using quantitative real-time PCR and whole mount in situ hybridization and found that tnfaip1 was highly expressed in early embryonic development and, subsequently, expression became localized to anterior embryonic structures. To investigate the function of tnfaip1 during early development, we constructed a model of a stably inherited tnfaip1 mutant using the CRISPR/Cas9 system. Tnfaip1 mutant embryos showed significant developmental delays as well as microcephaly and microphthalmia. At the same time, we found decreased expression of the neuronal marker genes tuba1b, neurod1, and ccnd1 in tnfaip1 mutants. Analysis of transcriptome sequencing data revealed altered expression of the embryonic development related genes dhx40, hspa13, tnfrsf19, nppa, lrp2b, hspb9, clul1, zbtb47a, cryba1a, and adgrg4a in the tnfaip1 mutants. These findings suggest an important role for tnfaip1 in the early development of zebrafish.

ARHGAP11A Is a Novel Prognostic and Predictive Biomarker Correlated with Immunosuppressive Microenvironment in Clear Cell Renal Cell Carcinoma.

Clear cell renal cell carcinoma (ccRCC) is a highly immunogenic tumor and immune dysfunction is associated with ccRCC poor prognosis. The RhoGTPase-activating proteins (RhoGAPs) family was reported to affect ccRCC development, but its role in immunity and prognosis prediction for ccRCC remain unknown. In the current study, we found ARHGAP11A was the only independent risk factor among 33 RhoGAPs (hazard ratio [HR] 1.949, 95% confidence interval [CI] 1.364-2.785). High ARHGAP11A level was associated with shorter overall survival (OS, HR 2.040, 95% CI 1.646-3.417) and ARHGAP11A is a prognostic biomarker for ccRCC. ARHGAP11A knockdown suppressed renal cell carcinoma (RCC) cell proliferation, colony formation, and migration, suggesting the promoting role of ARHGAP11A on RCC development. Mechanistically, ARHGAP11A might contribute to the suppressive tumor immune microenvironment (TIME). High ARHGAP11A level was correlated with infiltration of immunosuppressive cells (including T helper 2 (Th2) cells, regulatory T (Treg) cells, myeloid derived suppressor cells (MDSC), and M2 macrophage cells), activation of immunosuppressive pathways (IL6-JAK-STAT3 signaling and IFNγ response), and expression of inhibitory immune checkpoints (ICs). ARHGAP11A could promote T cell exhaustion and induce immune escape. ccRCC patients with low ARHGAP11A level were more suitable for immune checkpoint inhibitors (ICIs) therapy, while those with high ARHGAP11A level might benefit from a combination of ARHGAP11A blockade and ICIs. In all, ARHGAP11A might serve as a novel prognostic marker, therapeutic target, and predictor in the clinical response to ICIs therapy for ccRCC.

Deciphering the internal driving mechanism of microbial community for carbon conversion and nitrogen fixation during food waste composting with multifunctional microbial inoculation.

In this study, the effects of multifunctional microbial inoculation on food waste composting based on the synergistic property between organic matter degradation and nitrogen fixation were investigated. The results showed that inoculation simultaneously strengthened organic matter degradation by 9.9% and improved the nitrogen content by 20.6% compared with that of the control group. Additionally, spectral analysis demonstrated that inoculation was conducive to the enhanced humification, which was supported by the improvement in polyphenol oxidase activity. Microbial analysis showed that most of the introduced microorganisms (Bacillus, Streptomyces, Saccharomonospora) successfully colonized, and stimulated the growth of other indigenous microorganisms (Enterobacter, Paenibacillus). Meanwhile, the change in microbial community structure was accompanied by the enhanced tricarboxylic acid cycle and amino acid metabolism. Furthermore, network analysis and structural equation model revealed that the enhanced cooperation of microorganisms, in which more carbon sources could be provided by cellulose decomposition for nitrogen fixation.

Insights into the effect of iron-carbon particle amendment on food waste composting: Physicochemical properties and the microbial community.

The effects of iron-carbon (Fe-C) particle amendment on organic matter degradation, product quality and functional microbial community in food waste composting were investigated. Fe-C particles (10%) were added to the material and composted for 32 days in a lab-scale composting system. The results suggested that Fe-C particle enhanced organic matter degradation by 12.3%, particularly lignocellulose, leading to a greater humification process (increased by 15.5%). In addition, NO3--N generation was enhanced (15.9%) by nitrification with more active ammonia monooxygenase and nitrite oxidoreductase activities in the cooling and maturity periods. Fe-C particles not only significantly increased the relative abundances of Bacillus and Aspergillus for organic matter decomposition, but also decreased the relative abundances of acid-producing bacteria. RDA analysis demonstrated that the bacterial community was significantly influenced by dissolved organic matter, C/N, NO3--N, humic acid, volatile fatty acids and pH, while electrical conductivity was the key factor affecting the fungal community.

Cistanche extracts ameliorates the neurotoxicity induced by hydrogen peroxide in new mutant DJ-1-transfected neuroblastoma cellular models.

INTRODUCTION: DJ-1 mutation is a causative reason for familial Parkinson's disease (PD). Leucine166Proline (L166P) and C106S are two important DJ-1 mutations. In this study, we established hydrogen peroxide (H2 O2 ) induced L166P and C106S DJ-1-transfected neuroblastoma (SH-SY5Y) cellular models of PD and investigated the effects of Cistanche extracts and key bioactive compounds, including acteoside, echinacoside, caffeic acid, and Cistanche total glycosides on these two models. METHODS: After expressing FLAG-tagged L166P and C106S DJ-1 plasmids in Escherichia coli, the expressed plasmids were collected, treated with restriction enzyme, and identified using DNA electrophoresis. After purification, the L166P DJ-1 and C106S DJ-1 plasmids were separately transfected into SH-SY5Y cells using liposomes. Transfected SH-SY5Y cells were detected by western blotting and immunocytochemistry. Cell viability was determined using MTT assay. RESULTS: Both western blotting and immunocytochemistry showed that L166P and C106S DJ-1 were highly expressed in the transfected SH-SY5Y cells. MTT assays showed that transfection with L166P or C106S DJ-1 reduced the viability of SH-SY5Y cells exposed to H2 O2 , as compared to untransfected SH-SY5Y cells. In addition, Cistanche extracts and key bioactive compounds, including acteoside, echinacoside, caffeic acid, and Cistanche total glycosides, significantly inhibited the decreases of cell viability caused by H2 O2 in L166P and C106S DJ-1-transfected SH-SY5Y cells. CONCLUSIONS: These findings suggest that we successfully established sensitive and stable H2 O2 induced L166P DJ-1- and C106S DJ-1-transfected SH-SY5Y cell models of PD and Cistanche extracts may thus be useful for treating PD.

Correction: Se2Mo10V3, a heteropoly compound containing selenium, inhibits tumor growth.

[This corrects the article DOI: 10.18632/oncotarget.18909.].

Expression of the DJ-1 protein in the serum of Chinese patients with Parkinson's disease.

DJ-1 is one of the important genes found in Parkinson's disease (PD). Studies have shown that the DJ-1 protein levels are elevated in the cerebrospinal fluid (CSF) and plasma of sporadic PD patients, and the DJ-1 protein levels in the CSF and plasma may serve as biomarkers of PD. However, Japanese scholars previously reported that there was no difference in the levels of the DJ-1 protein in serum between sporadic PD patients and controls. Therefore, whether the serum DJ-1 protein levels are different between PD patients and controls in Chinese patients as well as whether serum DJ-1 protein can serve as a biomarker of PD are unknown. The present study aimed to determine whether there was a difference in serum DJ-1 protein levels between Chinese PD patients and controls. The subjects included 18 primary PD patients and 7 controls. Blood was collected by venipuncture, and serum was collected by centrifugation after the blood was coagulated. The serum DJ-1 protein levels were detected by both Western blot and ELISA. There were differences in the serum DJ-1 protein levels among different individuals. The serum DJ-1 concentration in PD patients was 11.3±10.1ng/ml, and that in controls was 18.1±12.8ng/ml (P>0.05). In conclusion, similar to the study conducted by Japanese scholars, we found no significant difference in the serum DJ-1 protein levels between PD patients and controls in Chinese subjects. The levels of the DJ-1 protein in serum may not be a biomarker of PD. In addition, there may be differences in the serum DJ-1 protein levels between Chinese and Japanese patients.

Se2Mo10V3, a heteropoly compound containing selenium, inhibits tumor growth.

Selenium compounds have strong anti-tumor effects and are well-tolerated. We examined the anti-tumor effects of (NH4)2H15Se2VIMo10V3O52·2H2O (Se2Mo10V3), a heteropoly compound containing selenium. Se2Mo10V3 inhibited proliferation in K562 cells with a half-maximal inhibitory concentration of 78.72±2.82 mg/L after 48 h and 24.94±0.88 mg/L after 72 h. Typical apoptotic morphologies were also observed in K562 cells treated with Se2Mo10V3, as were increased intracellular levels of Ca2+, Mg2+, H+, and reactive oxygen species, and decreased mitochondrial membrane potential. In addition, Se2Mo10V3 treatment triggered cytochrome C release and inhibited IκBα degradation and NF-κB translocation. In vivo experiments revealed that 5 or 10 mg/kg Se2Mo10V3 inhibited the growth of sarcoma 180 and hepatoma 22 xenograft tumors. These results indicate that Se2Mo10V3 inhibits tumor growth both in vitro and in vivo and induces apoptosis in K562 cells, possibly by inhibiting the NF-κB/IκBα pathway.

Protocatechuic acid inhibits rat pheochromocytoma cell damage induced by a dopaminergic neurotoxin.

Parkinson's disease (PD) is characterized by the progressive degeneration of dopaminergic neurons in the substantia nigra (SN) with the presence of alpha-synuclein inclusions termed Lewy bodies. The aggregation of alpha-synuclein into oligomeric species affects neuronal viability, having a causal role in the development of PD. The neuroprotective effects of protocatechuic acid (PAc) have been reported. However, the effects of PAc on tyrosine hydroxylase (TH) and alpha-synuclein in rat pheochromocytoma (PC12) cells treated with 1-methyl-4-phenylpyridinium ion (MPP(+)) remains unclear. In this study, we demonstrated that PAc inhibited the cytotoxicity, apoptotic morphology, reduction of TH expression and abnormal oligomeration of alpha-synuclein in PC12 cells treated with MPP(+). Taken together, our results indicate that the neuroprotective effects of PAc on PC12 cells treated with MPP(+) is related to the inhibition of the oligomerization of alpha-synuclein.