Objective: To construct a Family with sequence similarity 50 member A(FAM50A) gene knockout mouse insulinoma pancreatic β-cell line Beta-TC-6 using CRISPR/Cas9 gene editing technology and to prepare polyclonal antibodies specifically recognizing FAM50A. Methods: Two guide RNAs(sgRNAs) targeting the FAM50A gene were designed,and a recombinant plasmid expressing blue fluorescent protein(BFP) was constructed for gene knockout.The successfully constructed plasmid was transfected into Beta-TC-6 cells,and BFP-positive single cells were isolated for clonal expansion.The expanded monoclonal cell lines were genotyped by Sanger sequencing,and FAM50A protein expression was assessed by Western blot.Purified human recombinant FAM50A protein was used to immunize New Zealand rabbits for the preparation of a polyclonal antibody.The specificity of the prepared antibody was then validated using the successfully established FAM50A knockout cell line. Results: A monoclonal cell line with a successful knockout of the FAM50A gene was identified.Sanger sequencing confirmed base deletions at the target site.Western blot analysis showed a complete absence of FAM50A protein expression in this cell line.The prepared polyclonal antibody successfully recognized endogenous murine FAM50A protein in wild-type Beta-TC-6 cells and in hTERT-RPE1 cells overexpressing human FAM50A-GFP fusion protein,while no signal was detected in the FAM50A knockout cells. Conclusion This study successfully established a FAM50A gene knockout Beta-TC-6 cell model and generated a FAM50A polyclonal antibody,providing powerful tools for future research.

Pinelliae Rhizoma (PR), known as Banxia in Chinese, Hange in Japanese, and Banha in Korean, is a renowned herbal medicine in East Asia derived from the dry tuber of Pinellia ternata (Thunb.) Breit. (PT). It is extensively utilized in dispensing granules, classical prescriptions, and herbal formulas to treat various conditions, including cough, infection, phlegm, nausea, asthma, and inflammation. Despite numerous studies on PR and its classical prescriptions over recent decades, a comprehensive synthesis of available evidence regarding its multifunctional roles and therapeutic potential is lacking. This review aims to address this gap by examining emerging evidence from metabonomics, preclinical studies, and clinical trials, while exploring potential trends and prospects for future research. A systematic literature search was conducted across six electronic databases, including PubMed, Web of Science, Scopus, ScienceDirect, Wanfang, and China National Knowledge Infrastructure, to identify relevant articles on PR published until March 2023. PR contains 107 compounds with diverse pharmacological activities, including anti-inflammatory, immune regulatory, anti-viral, anti-cancer, anti-asthma, antitussive and expectorant, antioxidant, anti-obesity, anti-atherosclerosis, anti-microbial, emetic and anti-emetic, anti-convulsant and anti-epileptic, sedative and hypnotic, learning and memory enhancement, and anti-depressant effects. Metabonomic studies suggest that raw PR may exhibit cardiotoxicity and pregnancy toxicity while showing no apparent hepatorenal toxicity. However, limited pharmacokinetic investigations on PR constrain its clinical translation. Furthermore, clinical safety data on PR is scarce, with only four clinical trials assessing its positive effects in pediatric epilepsy, nausea and vomiting, soft tissue injury, and chronic sinus tract. This review aims to enhance understanding of PR and provide valuable information and recommendations for further research and development of herbal medicine.
Humans ; Pinellia/chemistry* ; Animals ; Ethnopharmacology ; Drugs, Chinese Herbal/chemistry* ; Phytochemicals/chemistry* ; Rhizome/chemistry*

Objective:To investigate the effects of Yangxue Qingnao granules combined with oxiracetam on cognitive function, psychiatric symptoms, and levels of inflammatory factors in patients with Alzheimer's disease. Methods:This study used a prospective design and included 100 patients with Alzheimer's disease who received treatment at the Department of Neurology, Shangluo Traditional Chinese Medicine Hospital, from January 2021 to June 2023. The patients were randomly divided into two groups: a control group and an observation group ( n = 50 per group). The control group received oxiracetam treatment, while the observation group received Yangxue Qingnao granules in addition to oxiracetam. All patients underwent treatment for 12 weeks and were followed up for 24 weeks. Levels of inflammatory factors (C-reactive protein, amyloid-β1-42, interleukin σ), oxidative stress responses (superoxide dismutase, malondialdehyde, glutathione peroxidase, heme oxygenase-1), psychiatric symptoms (assessed by the Behavioral Pathology in Alzheimer's Disease Rating Scale), and cognitive function (measured using the Alzheimer's Disease Assessment Scale-Cognitive Subscale and the Mini-Mental State Examination) were compared between the two groups before and after treatment. The response rate was also compared between the two groups. Data were statistically analyzed using SPSS 24.0, using t-tests for measurement data and chi-square tests for categorical data. Results:After treatment, the serum levels of C-reactive protein, amyloid-β1-42, interleukin σ, superoxide dismutase, and malondialdehyde as well as Alzheimer's Disease Assessment Scale-Cognitive Subscale and Behavioral Pathology in Alzheimer's Disease Rating Scale scores in the observation group were (2.50 ± 0.31) mg/L, (83.77 ± 19.77) ng/L, (80.10 ± 30.11) ng/L, (55.10 ± 4.42) kU/L, (3.33 ± 0.32) μmol/L, (13.26 ± 2.62), and (5.86 ± 1.14), respectively. All of these values were significantly lower than those in the control group [(4.53 ± 0.50) mg/L, (109.31 ± 24.19) ng/L, (148.53 ± 42.00) ng/L, (63.51 ± 4.25) kU/L, (4.87 ± 0.24) μmol/L, (17.90 ± 3.35), (10.16 ± 2.49), t = 24.40, -5.78, 9.36, 9.69, -27.22, -7.71, 11.10, all P<0.01]. The serum levels of glutathione peroxidase and heme oxygenase-1 as well as Mini-Mental State Examination scores in the observation group were (95.21 ± 5.43) U, (14.33 ± 1.58) μg/L, and (26.40 ± 1.11), respectively, which were significantly higher than those in the control group [(80.24 ± 5.33) U, (10.87 ± 1.24) μg/L, (23.53 ± 3.32) points, t = 13.91, 12.18, 5.79, all P<0.01]. Conclusions:Yangxue Qingnao granules combined with oxiracetam shows positive effects on improving behavioral symptoms, cognitive function, and overall clinical efficacy in patients with Alzheimer's disease.

Objective:To investigate the effects of Yangxue Qingnao granules combined with oxiracetam on cognitive function, psychiatric symptoms, and levels of inflammatory factors in patients with Alzheimer's disease. Methods:This study used a prospective design and included 100 patients with Alzheimer's disease who received treatment at the Department of Neurology, Shangluo Traditional Chinese Medicine Hospital, from January 2021 to June 2023. The patients were randomly divided into two groups: a control group and an observation group ( n = 50 per group). The control group received oxiracetam treatment, while the observation group received Yangxue Qingnao granules in addition to oxiracetam. All patients underwent treatment for 12 weeks and were followed up for 24 weeks. Levels of inflammatory factors (C-reactive protein, amyloid-β1-42, interleukin σ), oxidative stress responses (superoxide dismutase, malondialdehyde, glutathione peroxidase, heme oxygenase-1), psychiatric symptoms (assessed by the Behavioral Pathology in Alzheimer's Disease Rating Scale), and cognitive function (measured using the Alzheimer's Disease Assessment Scale-Cognitive Subscale and the Mini-Mental State Examination) were compared between the two groups before and after treatment. The response rate was also compared between the two groups. Data were statistically analyzed using SPSS 24.0, using t-tests for measurement data and chi-square tests for categorical data. Results:After treatment, the serum levels of C-reactive protein, amyloid-β1-42, interleukin σ, superoxide dismutase, and malondialdehyde as well as Alzheimer's Disease Assessment Scale-Cognitive Subscale and Behavioral Pathology in Alzheimer's Disease Rating Scale scores in the observation group were (2.50 ± 0.31) mg/L, (83.77 ± 19.77) ng/L, (80.10 ± 30.11) ng/L, (55.10 ± 4.42) kU/L, (3.33 ± 0.32) μmol/L, (13.26 ± 2.62), and (5.86 ± 1.14), respectively. All of these values were significantly lower than those in the control group [(4.53 ± 0.50) mg/L, (109.31 ± 24.19) ng/L, (148.53 ± 42.00) ng/L, (63.51 ± 4.25) kU/L, (4.87 ± 0.24) μmol/L, (17.90 ± 3.35), (10.16 ± 2.49), t = 24.40, -5.78, 9.36, 9.69, -27.22, -7.71, 11.10, all P<0.01]. The serum levels of glutathione peroxidase and heme oxygenase-1 as well as Mini-Mental State Examination scores in the observation group were (95.21 ± 5.43) U, (14.33 ± 1.58) μg/L, and (26.40 ± 1.11), respectively, which were significantly higher than those in the control group [(80.24 ± 5.33) U, (10.87 ± 1.24) μg/L, (23.53 ± 3.32) points, t = 13.91, 12.18, 5.79, all P<0.01]. Conclusions:Yangxue Qingnao granules combined with oxiracetam shows positive effects on improving behavioral symptoms, cognitive function, and overall clinical efficacy in patients with Alzheimer's disease.

Chemoresistance to 5-fluorouracil (5-FU) is a significant challenge in treating colorectal cancer (CRC). Novel combined regimens to thwart chemoresistance are therefore urgently needed. Herein, we demonstrated that the combination of Avenanthramide A (AVN A) and 5-FU has significant therapeutic advantages against CRC. Mechanistically, AVN A directly binds to the S198 site of the histone lysine demethylase KDM4C to promote its degradation, which subsequently fosters H3K9me3 occupancy on the MIR17HG promoter to block its transcription and derepress Bim expression. AVN A enhanced the therapeutic efficacy of 5-FU via impairing the KDM4C/MIR17HG/GSK-3β negative feedback loop. Importantly, the clinical correlation of the KDM4C/MIR17HG/Bim signaling axis with 5-FU response was validated in the refractory CRC patients. We provide evidence for the enhanced effectiveness of 5-FU when combined with AVN A in chemoresistant xenografts, CRC organoids, and Apc ^Min/+ mouse model. Additionally, AVN A mitigated the systemic adverse effects of 5-FU. Overall, our findings demonstrate that combinatorial therapy with AVN A and 5-FU represents an appealing opportunity and highlights KDM4C/MIR17HG/GSK-3β negative feedback loop which confers therapeutically exploitable vulnerability to chemo-refractory CRC patients.

Objective:To analyze the efficacy of unilateral arytenoid chondroplasty combined with minimally invasive mucosal flap plasty in the management bilateral vocal cord paralysis.Methods:A total of 66 patients with bilateral vocal cord paralysis hospitalized in the First Hospital of Sun Yat-sen University from January 2018 to December 2023 were retrospectively analyzed, among whom there were 8 males and 58 females, with ages ranging from 35 to 86 years old(mean age (57.8±11.6) years). All patients underwent suspension laryngoscopic CO 2 laser unilateral arytenoid chondroplasty and mucosal flap under general anesthesia. Postoperative follow-up period extended from 6 months to 6 years, with a median duration of 28 months. The study compared the degree of dyspnea and voice quality (subjective and objective evaluation) of the patients pre- and post-operatively, and analyzed the clinical differences between patients with and without preoperative or intraoperative tracheotomy, the extubation rate of tracheotomized patients, the recurrence rate, and the complication rate. Continuous variables conforming to normal distribution were tested by t-test and categorical variables by χ2 test,the Wilcoxon rank-sum test was used to analyze the improvement in dyspnea before and after surgery. Results:Compared with the preoperative period, 59 patients showed improvement in postoperative dyspnea ( U=161.5, P<0.01); there was no significant difference in voice disorder index 10, subjective auditory-perceptual assessment, and maximal vocalization time ( P>0.05), all of which remained within the relative normal range. Tracheotomy was performed in 32 out of 66 patients, with predominantly degree Ⅲ- dyspnea (46.9%, 15/32), including 26 patients with preoperative tracheotomy and 6 patients with intraoperative tracheotomy. Among the 34 patients who did not undergo tracheotomy, the majority presented with degree Ⅱ-dyspnea (82.4%, 28/34). All patients achieved successful extubation following surgery, with a mean median time to extubation of 1 month. Conclusions:The combination of unilateral arytenoid chondroplasty and minimally invasive mucosal flap plasty represents a refined and effective therapeutic approach for bilateral vocal cord paralysis. This minimally invasive technique has the potential to reduce the rate of tracheotomy.

Objective    To investigate the effects of telmisartan on the proliferation, migration and apoptosis of non-small cell lung cancer A549 and the mechanism of regulating Wnt signaling pathway. Methods    Non-small cell lung cancer cell line A549 was cultured in vitro. Cell counting kit-8 (CCK-8) assay was used to detect the effect of telmisartan at different concentrations on the proliferative activity of A549 cells. The survival fraction of A549 treated with different concentrations of telmisartan was determined by colony-formation assay. The effect of telmisartan at different concentrations on the migration ability of A549 cells was examined in the wounding healing assay. Hoechst staining was used to detect the effects of telmisartan at different concentrations on the apoptosis of A549. Western bloting was used to detect the expressions of β-actin, proliferating cell nuclear antigen (PCNA), Bax, Bcl-2, Wnt-3a, Beta-catenin (β-catenin), serine protein kinase 3β (p-GSK-3β), glycogen synthase kinase-3β (GSK-3β) and c-myc. Results    Different concentrations of telmisartan treatment inhibited the proliferation activity, colony-formation rate and migration of A549 cells, and reduced the expression of PCNA in a concentration-dependent manner. Telmisartan treatment promoted the apoptosis of A549 cells, significantly increased the expression of pro-apoptotic protein Bax and decreased the expression of anti-apoptotic protein Bcl-2. The expression levels of Wnt-3a, β-catenin, p-GSK-3β, and c-myc in A549 cells increased after treatment with telmisartan, while the expression levels of GSK-3β decreased. Conclusion    Telmisartan may play a role in the proliferation, migration and apoptosis of non-small cell lung cancer A549 cells, and  inhibiting the Wnt/β-catenin signaling pathway may be one of the mechanisms.

A new electrochemical sensor for organophosphate pesticide(methyl-paraoxon)detection based on bifunctional cerium oxide(CeO2)nanozyme is here reported for the first time.Methyl-paraoxon was degraded into p-nitrophenol by using CeO2 with phosphatase mimicking activity.The CeO2 nanozyme-modified electrode was then synthesized to detect p-nitrophenol.Cyclic voltammetry was applied to investigate the electrochemical behavior of the modified electrode,which indicates that the signal enhancement effect may attribute to the coating of CeO2 nanozyme.The current research also studied and discussed the main parameters affecting the analytical signal,including accumulation potential,accumulation time,and pH.Under the optimum conditions,the present method provided a wider linear range from 0.1 to 100 μmol/L for methyl-paraoxon with a detection limit of 0.06 μmol/L.To validate the proof of concept,the electrochemical sensor was then successfully applied for the determination of methyl-paraoxon in three herb samples,i.e.,Coix lacryma-jobi,Adenophora stricta and Semen nelum-binis.Our findings may provide new insights into the application of bifunctional nanozyme in electro-chemical detection of organophosphorus pesticide.

Objective: To assess the efficacy of supraclavicular artery island flap (SCAIF) for the reconstruction of hypopharynx and upper esophagus. Methods: The SCAIF procedure on was used in 10 patients, including 8 with hypopharygeal carcinomas, 1 with esophageal carcinoma and neck skin invasion and 1 with hypopharyngeal leiomyosarcoma, at the Otorhinolaryngology Hospital, the First Affiliated Hospital, Sun Yatsen University between December 2015 and June 2017. The sizes of the flaps were measured in (4-8) cm×(5-12) cm. Clinical indexes such as harvesting time and survival were recorded. Results: Harvesting time for SCAIF ranged from 20 to 30 minutes, averaging 26 minutes. Nine flaps survived, one flap had partially necrosis. Functional outcomes were excellent and the donor sites were direct closed without complications. Conclusions SCAIF is a versatile, reliable, and easily harvested flap, with good cosmetic and functional outcomes for reconstructing the defects of hypopharynx and upper esophagus.

ATP-Binding Cassette Transporters ; genetics ; DNA Copy Number Variations ; genetics ; Genetic Predisposition to Disease ; Gout ; genetics ; Humans ; Receptors, IgG ; genetics ; Receptors, Interleukin-17 ; genetics