Xiaoaiping (XAP) Injection demonstrates the anti-prostate cancer (PCa) effects, yet the underlying mechanism remains unclear. This study aims to investigate the impact of XAP on PCa and elucidate its mechanism of action. PCa cell proliferation was evaluated using a cell counting kit-8 (CCK-8) assay. Cell apoptosis was assessed through Hoechst staining and Western blotting assays. Proteomics technology was employed to identify key molecules and significant signaling pathways modulated by XAP in PCa cells. To further validate potential key genes and important pathways, a series of assays were conducted, including acridine orange (AO) staining, transmission electron microscopy, and immunofluorescence assays. The molecular mechanism of XAP against PCa in vivo was examined using a PC3 xenograft mouse model. Results demonstrated that XAP significantly inhibited cell proliferation in multiple PCa cell lines. In C4-2 and prostate cancer cell line-3 (PC3) cells, XAP induced cellular apoptosis, evidenced by reduced B-cell lymphoma 2 (Bcl-2) levels and elevated Bcl-2-associated X (Bax) levels. Proteomic, immunofluorescence, and quantitative reverse transcription-polymerase chain reaction (qRT-PCR) investigations revealed a strong correlation between forkhead box O3a (FoxO3a) autophagic degradation and the anti-PCa action of XAP. XAP hindered autophagy by reducing the expression levels of autophagy-related protein 5 (Atg5)/autophagy-related protein 12 (Atg12) and enhancing FoxO3a expression and nuclear translocation. Furthermore, XAP exhibited potent anti-PCa action in PC3 xenograft mice and triggered FoxO3a nuclear translocation in tumor tissue. These findings suggest that XAP induces PCa apoptosis via inhibition of FoxO3a autophagic degradation, potentially offering a novel perspective on XAP injection as an effective anticancer therapy for PCa.
Male ; Humans ; Prostatic Neoplasms/physiopathology* ; Autophagy/drug effects* ; Animals ; Drugs, Chinese Herbal/pharmacology* ; Proteomics ; Mice ; Apoptosis/drug effects* ; Cell Line, Tumor ; Cell Proliferation/drug effects* ; Forkhead Box Protein O3/genetics* ; Xenograft Model Antitumor Assays ; Mice, Nude ; Mice, Inbred BALB C

This study established the mouse podocyte clone-5 (MPC5) with Smad3 knockout and studied the effect of transforming growth factor-beta 1 (TGF-β1) on the dedifferentiation of the MPC5 cells with Smad3 knockout, aiming to provide a cell tool for studying the role of Smad3 in mouse podocytes. The single-guide RNA (sgRNA) sequence targeting Smad3 was designed according to the principles of CRISPR/Cas9 design. The pX458-Smad3 vector was constructed and introduced into competent cells, and then the vector was extracted and used to transfect MPC5 cells. The successfully transfected cells were sorted by a flow cytometer. After single-cell clone expansion, PCR amplification of sequences adjacent to the edition site of Smad3 and sequencing were performed to identify potential cells with gene knockout. Western blotting was employed to verify the knockout efficiency of Smad3. Finally, the effect of Smad3 knockout on TGF-β1-induced dedifferentiation of MPC5 cells was analyzed by reverse transcription-polymerase chain reacting (RT-PCR), Western blotting, and the immunofluorescence method. The sgRNA was designed to target the fifth exon of Smad3. EGFP expression was observed 24 h after transfection of the pX458-Smad3 plasmid into MPC5 cells, with the transfection efficiency of 0.1% as determined by flow cytometry. From the transfected cells, 21 cell clones were obtained through flow cytometric sorting and single-cell clone expansion. PCR amplification and sequencing of the region around the sgRNA target site in Smad3 identified two cell clones with biallelic frameshift mutations. Western blotting results confirmed the absence of Smad3 expression in these clones, indicating successful establishment of the MPC5 cell line with Smad3 knockout. In normal MPC5 cells, TGF-β1 stimulation promoted the expression of fibrosis-related genes fibronectin and Col1a1 (collagen I) and inhibited the expression of the podocyte marker proteins synaptopodin and podocin, which suggested epithelial-mesenchymal transition and podocyte injury. However, in the two MPC5 cell lines with Smad3 knockout, TGF-β1-induced expression of epithelial-mesenchymal transition markers was significantly suppressed. The MPC5 cell lines with Smad3 knockout that were constructed by CRISPR/Cas9 provide a valuable cell model for functional studies of Smad3 protein and highlight the critical role of Smad3 in cell dedifferentiation.
Animals ; Smad3 Protein/genetics* ; CRISPR-Cas Systems/genetics* ; Mice ; Podocytes/metabolism* ; Transforming Growth Factor beta1/pharmacology* ; Cell Line ; Gene Knockout Techniques ; RNA, Guide, CRISPR-Cas Systems/genetics*

Porcine epidemic diarrhea virus (PEDV) is an intestinal coronavirus that can cause porcine epidemic diarrhea, leading to diarrhea, vomiting, weight loss, and even death in piglets. Due to the diversity of PEDV strains, traditional vaccines are difficult to sustainably and effectively prevent and control PEDV. This article reviews the strategies and mechanisms of active substances in regulating intracellular signaling pathways, viral proteins, and microbial metabolites to enhance the host immune function against PEDV. It emphasizes the prevention of PEDV resistance and the potential harm of PEDV breaking through interspecies barriers to the human society, aiming to provide reliable theoretical support for the development of new antiviral drugs or vaccines.
Porcine epidemic diarrhea virus/immunology* ; Animals ; Swine ; Swine Diseases/prevention & control* ; Antiviral Agents/pharmacology* ; Coronavirus Infections/virology* ; Viral Vaccines/immunology* ; Humans ; Signal Transduction

The control strategy of rehabilitation robots should not only adapt to patients with different levels of motor function but also encourage patients to participate voluntarily in rehabilitation training.However,existing control strategies usually consider only one of these aspects.This study proposes a voluntary and adaptive control strategy that solves both questions.Firstly,the controller switched to the corresponding working modes(including challenge,free,assistant,and robot-dominant modes)based on the trajectory tracking error of human-robot cooperative movement.To encourage patient participation,a musculoskeletal model was used to estimate the patient's active torque.The robot's output torque was designed as the product of the active torque and a coefficient,with the coefficient adaptively changing according to the working mode.Experiments were conducted on two healthy subjects and four hemiplegic patients using an ankle rehabilitation robot.The results showed that this controller not only provided adaptive the robot's output torque based on the movement performance of patients but also encouraged patients to complete movement tasks themselves.Therefore,the control strategy has high application value in the field of rehabilitation.

Objective To investigate the role of LncRNA-gm33782 in tubular injury in acute kidney injury(AKI)and the mechanism by which isorhamnetin ameliorates inflammatory cell apoptosis of tubular cells in AKI.Methods Forty-eight male C57BL/6J mice were randomly assigned to four groups:control group,AKI model group,electroporation+AKI group,and treatment group(oral administration of 30 mg/kg isorhamnetin).AKI was induced by a one-time intraperitoneal injection of 20 mg/kg cisplatin.Chromatin isolation by RNA purification was used to capture the LncRNA-gm33782 binding protein in AKI-affected kidneys for mass spectrometry,revealing the direct target protein of LncRNA-gm33782.The role of LncRNA-gm33782 in AKI was evaluated by observing the renal function,pathological structure,and expression of renal inflammatory factors(interleukin-1β,interleukin-6,and tumor necrosis factor-α)in mice after electrotransfection and isorhamnetin treatment.Nuclear factor-κB was detected as a critical mediator of inflammation.The expression levels of Bax and Bcl-2 protein were detected and flow cytometry was performed to evaluate the therapeutic effect of isorhamnetin on tubular cell apoptosis in AKI.Results Kidneys with cisplatin-induced AKI showed severe renal tubule injury,macrophage infiltration,and inflammation.Isorhamnetin treatment and LncRNA-gm33782 electrotransfection knockdown alleviated these signs of AKI.LncRNA-gm33782 was mainly expressed in renal tubular cells with AKI.LncRNA-gm33782 binding protein was detected by mass spectrometry,and complement factor H was found to have a direct binding relationship with LncRNA-gm33782.After LncRNA-gm33782 was overexpressed in vitro,the expression of complement factor-H immediately increased,and the therapeutic effect of isorhamnetin on apoptosis of AKI-affected inflammatory cells was inhibited.Conclusions Isorhamnetin alleviates apoptosis of tubular inflammatory cells in AKI by inhibiting the regulatory effect of LncRNA-gm33782 on complement factor H.

Objective To investigate the therapeutic effect and underlying mechanism of Qishishenshu Capsule on renal fibrosis in mice with early diabetic nephropathy(DN).Methods A DN mouse model was established by multiple injections of streptozotocin.The mice were randomly divided into a normal group(NC),model group(DN),and Qishi group(QS)(0.9 g/(kg·d)),with eight mice in each group.Mice were gavaged continuously for 4 weeks,and fasting blood glucose(FBG)was measured weekly.Four weeks later,urinary albumin/creatinine(UACR),serum creatinine,and blood urea nitrogen were measured.Hematoxylin-eosin,periodicacid-Schiff,and Sirius red staining were used to analyze renal pathological changes.Real-time fluorescence quantitative reverse-transcription polymerase chain reaction was used to detect the mRNA levels of fibronectin(FN),collagen type Ⅰ alpha 1(Col1a1),and α-smooth muscle actin(α-SMA).Immunohistochemistry and Western blot were performed to detect FN,collagen type Ⅰ(Collagen Ⅰ),collagen typeⅢ(Collagen Ⅲ),α-SMA,Podocin,Nephrin,and transforming growth factor-β1/SMAD family member2/3(TGF-β1/Smad2/3)pathway-related proteins.Results Compared with mice in the NC group,those in the DN group showed significantly higher levels of FBG and UACR(P＜0.001),and mesangial hyperplasia,basement membrane thickening,and collagen deposition in the renal tissue.The mRNA levels of FN,Col1a1,and α-SMA were increased(P＜0.05).Protein levels of Podocin and Nephrin were decreased(P＜0.05).The levels of FN,Collagen I,Collagen Ⅲ,α-SMA,and TGF-β1/Smad2/3 pathway proteins were increased(P＜0.05).Compared with the DN group,the QS group's level of UACR was decreased(P＜0.05),their renal pathological injury was alleviated,and mRNA levels of FN,Collagen Ⅰ,andα-SMA were attenuated(P＜0.05);whereas their protein levels of Podocin and Nephrin were elevated(P＜0.05).The levels of FN,Collagen Ⅰ,Collagen Ⅲ,α-SMA,and TGF-β1/Smad2/3 pathway proteins were also decreased(P＜0.05).Conclusions Qishishenshu Capsule improved renal fibrosis in DN mice,probably through the inhibition of the TGF-β1/Smad2/3 signaling pathway.

Objective:To understand the problems existing in the curriculum design of food hygiene and nutrition for undergraduates, and put forward countermeasures to improve the quality of talents cultivation and promote professional development.Methods:Self-made questionnaires were distributed by convenience sampling method, through online and offline ways, and the graduates' opinions on the curriculum design of their school. Quantitative data were analyzed by SPSS 26.0, and qualitative data were analyzed by NVivo 11.0.Results:A total of 258 valid questionnaires were recovered, and the average score of graduates' satisfaction with teaching quality was 2.42±1.02. Graduates had different opinions on the importance of special courses, and 150 students (58.14%) were satisfied with major courses. Some graduates thought the time of special courses was unreasonable and there were few major courses. Besides, 141 graduates (54.65%) were satisfied with the clinical probation in their graduation colleges. Although graduates believed that innovation ability was important, innovative education courses were scarce at present.Conclusions:Graduates are less satisfied with the setting of undergraduate curriculum in food hygiene and nutrition. It is suggested to optimize professional courses setting, improve experimental facilities and equipment, strengthen the management of practice bases, and pay attention to the cultivation of students' innovation and entrepreneurship ability.

Objective To assess the impact of intravenous esketamine administered prior to car-diopulmonary bypass(CPB)initiation on ventricular function and internal carotid artery blood flow in pa-tients undergoing heart valve replacement surgery.Methods Sixty patients underwent elective CPB heart valve replacement,38 males and 22 females,aged 18-75 years,BMI 18.5-30.0 kg/m2,ASA physical status Ⅱ or Ⅲ,NYHA cardiac function classification Ⅰ-Ⅲ,and a left ventricular ejection fraction(LVEF)of≥45％,were selected.The patients were randomly divided into two groups:esketamine group(group E)and normal saline group(group C),30 patients in each group.Total intravenous anesthesia was used during the operation.Following the initiation of CPB,group E received an intravenous infusion of es-ketamine at a rate of 0.5 mg·kg-1·h-1 until the conclusion of the procedure,while group C received an equivalent volume of normal saline concurrently at the same rate.HR,MAP,CVP,and cardiac output index(CI)were recorded before anesthesia induction,during skin resection,and within 60 minutes after stopping CPB.LVEF,left ventricular global longitudinal strain(GLS),global longitudinal time-to-peak strain standard deviation(GLTSD),global circumferential strain(GCS),global circumferential time-to-peak strain standard deviation(GCTSD),right ventricular ejection fraction(RVEF),right ventricular GLS,and GLTSD were obtained during skin resection,within 40 minutes of CPB,and 60 minutes after stopping CPB.rScO2,BIS,concentrations of Hb and lactic acid(Lac),peak systolic flow velocity(SPV),quantity of flow-internal carotid artery(Q-ICA),and blood flow resistance index(RI)were recorded before anesthesia induction,during skin resection,within 40 minutes of CPB,and within 60 minutes after stopping CPB.Concentrations of cardiac troponin Ⅰ(cTnⅠ),alanine aminotransferase(ALT),creatinine(Cr),and neuron-specific enolase(NSE)were recorded before anesthesia induction and 6 hours after operation.Spon-taneous resuscitation after CPB,postoperative extubation time,duration of ICU stay,total hospital stay,in-cidence of adverse cardiac events,and 30-day postoperative mortality were recorded.Results Compared with group C,group E exhibited a significant increase in CI within 60 minutes after stopping CPB(P＜0.05).The LVEF,RVEF,and right ventricular GLS demonstrated significant increases within 60 minutes after stopping CPB in group E compared with group C(P＜0.05).The left ventricular GLS and left ven-tricular GCTSD displayed significant increases 30 minutes after stopping CPB in group E compared with group C.The RI exhibited a significant increase within 40 minutes of CPB in group E compared with group C(P＜0.05).There were no significant differences in cTnⅠ,ALT,Cr,NSE,spontaneous resuscitation affter CPB,postoperative extubation time,duration of ICU stay,total hospital stay,incidence of cardiac adverse events,and 30-day postoperative mortality between the two groups.Conclusion Administration of esket-amine following the onset of CPB in patients undergoing cardiac surgery demonstrates a significant elevation in CI post-CPB cessation.Furthermore,it may augment ventricular longitudinal strain,thereby enhancing myocardial contraction,leading to increased postoperative ventricular ejection fraction,and sustaining hemo-dynamic stability.

Objective To investigate the role of HK2 and VDAC1 in diacetylmorphine-induced cardiomyocyte apoptosis.Methods A dose-escalation method was used to establish a rat model of diacetylmorphine addiction.Forty SD rats were randomly divided into three groups,the normal group(n=10)was injected with an equal amount of saline subcutaneously,the model group(n=15)was injected with 5 mg/kg of diacetylmorphine for the first time,and then the dose was increased by 2.5 mg/(kg·d)day by day for 20 days,and the group of model +10 D(n=15)continued to increase the dose based on the model group up to the 10th day.Lactate dehydrogenase(LDH)and glutamic oxaloacetic transaminase(GOT)were detected by ELISA;HE staining was used to observe the pathological changes of myocardial tissues in each group;TUNEL staining was used to detect apoptosis in myocardial tissues in each group;and immunohistochemistry,RT-q-analysis,and immunochemistry were used to detect apoptosis in myocardial tissues in each group.Immunohistochemistry,RT-qPCR and Western bl-ot were used to detect the mRNA and protein expression of HK2,VDAC1 and apoptosis-related factors.Results HE staining revealed that myocardial tissues exhibited different degrees of damage with the prolongation of diacetylmorphine intervention.Compared with the normal group,serum LDH,GOT content and myocardial apoptosis rate increased in the model group,mRNA and protein levels of HK2 and anti-apoptotic factor Bcl-2 decreased,mRNA and protein levels of VDAC1 and pro-apoptotic factors Bax and Caspase-3 increased,and the protein level of Clevead Caspase-3 increased;in the model +10 D group the above indexes,there was a statistically significant difference(P<0.05).Conclusion Diacetylmorphine can cause cardiomyocyte apoptosis,and VDAC1 may be involved in the process of cardiomyocyte apoptosis caused by diacetylmorphine.

Objective:To compare the efficacy of pecto-intercostal fascial plane (PIFP) block versus transversus thoracic muscle plane (TTP) block under ultrasound guidance in coronary artery bypass grafting with general anesthesia.Methods:Ninety American Society of Anesthesiologists Physical Status classification Ⅱor Ⅲ patients of either sex, aged 50-79 yr, scheduled for elective coronary artery bypass grafting, were divided into 3 groups ( n=30 each) using a random number table method: PIFP block combined with general anesthesia group (PG group), TTP block combined with general anesthesia group (TG group), and general anesthesia group (G group). After anesthesia induction, bilateral PIFP block was performed under ultrasound guidance in group PG, TTP block was performed under ultrasound guidance in group TG. Three groups used the same general anesthesia method and patient-controlled intravenous analgesia after surgery. Visual analog scale scores (cough, position change, etc) at rest and during activity were recorded at 6, 12, 18 and 24 h after operation. The total consumption of intraoperative sufentanil, extubation time, length of stay in intensive care units, rate of rescue analgesia, effective pressing times of patient-controlled analgesia, incidence of postoperative nausea and vomiting, skin pruritus and nerve block-related adverse events were recorded. The operation time of nerve block was recorded and ultrasound-guided needle visibility score was assessed in PG group and TG group. Results:Compared with group G, the total consumption of intraoperative sufentanil was significantly reduced, the extubation time and length of stay in intensive care units were shortened, visual analog scale scores at rest and during activity were decreased at 6, 12 and 18 h after operation, the rate of rescue analgesia was decreased, and the effective pressing times of patient-controlled analgesia were decreased in group PG and group TG ( P<0.05), and no significant change was found in the aforementioned parameters in PG and TG groups ( P> 0.05). Compared with group TG, the operational time of nerve block was significantly shortened, and the ultrasound-guided needle visibility score was increased in group PG ( P<0.05). No nerve block-related adverse events were found in PG and TG groups. There was no significant difference in the incidence of postoperative nausea and vomiting and skin pruritus among the three groups ( P>0.05). Conclusions:PIFP block can provide good perioperative analgesia and promote the rapid recovery in the patients undergoing coronary artery bypass grafting with general anesthesia. Although the analgesic effect of PIFP blockade is similar to that of TTP blockade, PIFP blockade is more clinically valuable due to its simpler operation and less relative risk.