Objective To investigate the effects of ischemia time and storage periods on RNA quality in fresh-frozen breast cancer(BC)and esophageal cancer(EC)tissue samples in order to establish evidence-based protocols for biobank sample management.Methods The tumor(T)and paired normal(N)tissue samples from 6 cases of BC and 6 cases of EC were collected and cryopreserved in Biobank,Shantou Central Hospital.Mirror paraffin-embedded tissues were simultaneously prepared into sections for morphological analysis.The samples were divided into two groups of<15 min and 15-30 min according to ischemia time,and RNA quality was analyzed at 4 storage periods of 8-10 months(T1),14-16 months(T2),26-28 months(T3)and 38-40 months(T4).Results In 96 analyzed samples,93.8%(90/96)exhibited high quality(RIN≥6),with 89.6%(43/48)in BC and 97.9%(47/48)in EC.Significant differences in RIN were observed between BC group and EC group(8.050 vs.8.600,P=0.009).In EC group,RIN value was significantly negatively correlated with RNA yield(P<0.001).Moreover,RIN values of tumor-normal pairs exhibited markedly significant differences(7.550 vs.9.000,P<0.001).In contrast,no significant difference was detected in BC group(8.200 vs.7.700,P=0.348).Statistical analysis showed that RIN value was positively correlated with 28S/18S(P<0.001),but had no correlation with tumor content(P=0.676)and necrotic content(P=0.055).Neither ischemia time(<15 min vs.15-30 min:8.200 vs.8.300,P=0.932)nor storage periods(T1-T4:8.400,7.700,8.450,8.600,P=0.163)compromised RNA quality.Conclusion Organ origin and tissue type could influence RNA quality of fresh-frozen tissue samples.However,limited ischemia time(≤30 min)and long-term storage period(38-40 months)do not adversely affect RNA quality in fresh-frozen breast cancer and esophageal cancer tissue samples.

Thyroid eye disease(TED), also known as thyroid-associated ophthalmopathy(TAO) or Graves' ophthalmopathy(GO), is an organ-specific autoimmune disorder associated with autoimmune thyroid disease and can severely impair vision and quality of life. The pathogenesis of TED is closely linked to dysregulated cytokine networks; however, current therapies yield limited response rates in patients with moderate-to-severe TED. This review synthesizes the mechanisms of action of key cytokines and evaluates the translational potential of emerging targeted therapies, aiming to provide a rationale for overcoming current therapeutic bottlenecks and advancing precision treatment strategies.

Objective:To evaluate the effectiveness of fosaprepitant in preventing postoperative nausea and vomiting (PONV) following gynecological laparoscopic surgery.Methods:In this randomized parallel-controlled trial, 100 American Society of Anesthesiologists Physical Status classification Ⅰ or Ⅱ patients, aged 18-64 yr, undergoing elective gynecological laparoscopic surgery under general anesthesia at the First Affiliated Hospital of Zhengzhou University, were selected and divided into 2 groups ( n=50 each) in a ratio of 1∶1 using blocked randomization: fosaprepitant group (group F) and tropisetron group (group T). At 30 min before anesthesia induction, fosaprepitant 150 mg was intravenously infused in group F, and tropisetron 5 mg was intravenously infused in group T, both diluted in 150 ml of normal saline. Anesthesia was induced by intravenous injection of midazolam, etomidate, sufentanil and cisatracurium. Anesthesia was maintained by intravenous infusion of remifentanil and propofol. Patient-controlled intravenous analgesia was performed with hydromorphone at the end of operation until 48 h after operation. Metoclopramide was given as rescue antiemetic. The PONV, requirement for antiemetic drugs and related adverse reactions were recorded within 24 h after surgery. Results:The incidence of PONV (10% vs 30%), the incidence of vomiting(2% vs 16%) and the rescue rate of antiemetic drugs(2% vs 12%)were significantly lower in group F than in group T ( P<0.05). There was no significant difference in the incidence of related adverse reactions between the two groups ( P>0.05). Conclusions:Intravenous infusion of fosaprepitant 150 mg at 30 min before anesthesia induction effectively prevents PONV in patients undergoing gynecological laparoscopic surgery, and the efficacy is superior to that of the conventional use of tropisetron.

Objective:To investigate the risk factors for positive surgical margins（PSM）after robot-assisted radical prostatectomy（RARP），and to develop and validate a predictive nomogram.Methods:We retrospectively analyzed the clinicopathological data of 874 prostate cancer patients who underwent RARP performed by a single surgeon at the First Medical Center of Chinese PLA General Hospital between January 2012 and December 2018. Patients were divided into positive surgical margin（n=327）and negative surgical margin（n=547）groups based on postoperative margin status.The PSM group had significantly higher preoperative median tPSA［31.200（19.050，54.400）ng/ml vs. 15.050（9.840，27.590）ng/ml， P<0.01］，higher proportion of patients with PSAD>1 ng/ml 2［49.5%（162/327）vs. 21.2%（116/547）， P<0.01］，biopsy Gleason score ≥8［33.3%（109/327）vs. 21.2%（116/547）， P<0.01］，ISUP grade 4-5［33.3%（109/327）vs. 21.2%（116/547）， P<0.01］，clinical T stage ≥cT 3［11.3%（37/327）vs. 4.2%（23/547）， P<0.01］，and high-risk classification［82.3%（269/327）vs. 55.9%（306/547）， P<0.01］compared to the negative surgical margin group. Conversely，the PSM group had a lower prevalence of hypertension［29.7%（97/327）vs. 40.2%（220/547）， P=0.002］.Patients were randomly split into a training cohort（n=656，75%）and an internal validation cohort（n=218，25%）. An external validation cohort included 71 patients who underwent RARP by different surgeons between January 2014 and December 2016. No significant differences in baseline characteristics were observed between cohorts（ P>0.05）.Univariate and multivariate logistic regression analyses identified independent predictors of PSM，which were incorporated into a nomogram. Predictive performance was assessed using receiver operating characteristic（ROC）curves，decision curve analysis（DCA），and calibration curve. Internal and external validations were performed. Results:The PSM group had longer postoperative hospitalization［6（5，8）vs. 6（5，7）days， P=0.028］，higher rates of pathologic Gleason score ≥8［41.5%（115/277）vs. 24.9%（111/446）， P<0.01］，ISUP grade 4-5［41.5%（115/277）vs. 24.9%（111/446）， P<0.01］，pT 3 stage［52.3%（171/327）vs. 17.4%（95/547）， P<0.01］，pN 1 stage［12.8%（42/327）vs. 3.8%（21/547）， P<0.01］，extracapsular extension［52.3%（171/327）vs. 17.4%（95/547）， P<0.01］，and seminal vesicle invasion［34.6%（113/327）vs. 9.1%（50/547）， P<0.01］.Multivariate analysis identified elevated tPSA（ OR=1.014，95% CI 1.004—1.024，P=0.006）and PSAD ≥0.15 ng/（ml/g）（ OR=11.638，95% CI 1.450—93.396，P=0.021）as independent risk factors for PSM. The area under the ROC curve（AUC）of the nomogram constructed based on the above variables was 0.770（95% CI 0.735—0.805）. The AUC values for the internal and external validation sets were 0.698（95% CI 0.630—0.767）and 0.643（95% CI 0.513—0.774），respectively. The calibration curve demonstrated good agreement between the predicted and observed outcomes，and the DCA indicated that the predictive model has potential clinical utility in decision-making. Conclusion:tPSA and PSAD were identified as independent risk factors for PSM. The nomogram constructed based on these two independent predictive variables effectively predicted PSM after RARP.

The scaphoid bone is one of the important bone of hand,which is frequently injured and difficult to treat in clinical practice.Therefore,it is very important to deeply study the microstructure and biomechanical characteristics of the scaphoid bone for understanding its injury mechanism and optimizing treatment scheme.Microcomputed tomography(micro-CT)provides high-resolution imaging of bone tissue,while finite element analysis can help to simulate the stress distribution and behavioral patterns of the scaphoid bone under various physiological and pathological states.The high-resolution three-dimensional image of the scaphoid bone obtained by micro-CT technology can be used to construct finite element models of real anatomical structure of the scaphoid bone,thus achieving accurate simulation of the mechanical properties of the scaphoid bone.The fusion of these two advanced technologies provides a new perspective for revealing the structural and functional relationships and injury mechanism of the scaphoid bone.Therefore,this paper reviews the anatomical characteristics of the scaphoid bone and its biomechanical behavior in different states,emphasizing the specific applications and advantages of micro-CT and finite element analysis techniques in the study of the scaphoid bone.By summarizing the research findings in recent years,this paper provides novel scientific basis and methods for the diagnosis,treatment,and prevention of scaphoid bone-related disorders.

Objective:To evaluate the effectiveness of fosaprepitant in preventing postoperative nausea and vomiting (PONV) following gynecological laparoscopic surgery.Methods:In this randomized parallel-controlled trial, 100 American Society of Anesthesiologists Physical Status classification Ⅰ or Ⅱ patients, aged 18-64 yr, undergoing elective gynecological laparoscopic surgery under general anesthesia at the First Affiliated Hospital of Zhengzhou University, were selected and divided into 2 groups ( n=50 each) in a ratio of 1∶1 using blocked randomization: fosaprepitant group (group F) and tropisetron group (group T). At 30 min before anesthesia induction, fosaprepitant 150 mg was intravenously infused in group F, and tropisetron 5 mg was intravenously infused in group T, both diluted in 150 ml of normal saline. Anesthesia was induced by intravenous injection of midazolam, etomidate, sufentanil and cisatracurium. Anesthesia was maintained by intravenous infusion of remifentanil and propofol. Patient-controlled intravenous analgesia was performed with hydromorphone at the end of operation until 48 h after operation. Metoclopramide was given as rescue antiemetic. The PONV, requirement for antiemetic drugs and related adverse reactions were recorded within 24 h after surgery. Results:The incidence of PONV (10% vs 30%), the incidence of vomiting(2% vs 16%) and the rescue rate of antiemetic drugs(2% vs 12%)were significantly lower in group F than in group T ( P<0.05). There was no significant difference in the incidence of related adverse reactions between the two groups ( P>0.05). Conclusions:Intravenous infusion of fosaprepitant 150 mg at 30 min before anesthesia induction effectively prevents PONV in patients undergoing gynecological laparoscopic surgery, and the efficacy is superior to that of the conventional use of tropisetron.

Aim To investigate the role and regulatory mechanism of CREB regulated transcription coactivator 2(CRTC2)in cardiomyocyte hypertrophy.Methods A pathological cardiomyocyte hypertrophy model was established in C57BL/6 mice by intraperitoneal injection of isoproterenol(ISO),the expression of CRTC2 in cardiac tissue was detec-ted by Western blot.The CRTC2 knockout mice model was constructed,the cardiac function of mice was detected by small animal echocardiography,the collagen fiber content in mice cardiac tissue was detected by Masson staining,the car-diomyocyte hypertrophy related proteins:skeletal muscle α1-actin(ACTA1)and brain natriuretic peptide(BNP),as well as ferroptosis related proteins:acyl-CoA synthetase long chain family member 4(ACSL4),solute carrier family 7 member 11(SLC7A11)and glutathione peroxidase 4(GPX4)in mice cardiac tissue were detected by Western blot,the iron ion content in mice cardiac tissue was detected by iron ion kit,to evaluate the correlation between CRTC2 and cardiomyocyte hypertrophy and ferroptosis.H9c2 cells were induced by ISO to construct an in vitro model of cardiomyocyte hypertrophy,the protein expressions of CRTC2,ACTA1,BNP,ACSL4,SLC7A11 and GPX4 were detected after intervention with fer-roptosis inhibitor ferrostatin-1(Fer-1).H9c2 cells with CRTC2 overexpression induced by ISO were used to construct an in vitro model of cardiomyocyte hypertrophy,the related indicators of cardiomyocyte hypertrophy and ferroptosis were detec-ted to explore the mechanism of CRTC2 in cardiomyocyte hypertrophy.Results Compared with the control group,the expression of CRTC2 protein in the cardiac tissue of ISO induced cardiomyocyte hypertrophy mice was increased(P＜0.05).Compared with wild-type mice,CRTC2-/-mice showed worsened cardiac function,manifested as increased left ventricular end-diastolic diameter(LVEDD),left ventricular end-systolic diameter(LVESD),left ventricular posterior wall thickness(LVPWT),heart weight/tibia length(HW/TL)and heart weight/body weight(HW/BW),decreased short axis shortening(FS)and ejection fraction(EF),increased collagen fiber content in cardiac tissue,upregulated ex-pression of cardiomyocyte hypertrophy-related proteins ACTA1 and BNP,increased mRNA and protein expression of ferrop-tosis-related protein ACSL4,decreased mRNA and protein expression of SLC7A11 and GPX4,and elevated iron ion content in cardiac tissue(P＜0.05 or P＜0.01).In vitro experiments showed that compared with ISO group,the ISO+Fer-1 group had no significant change in CRTC2 protein expression(P＞0.05),the expression of ACTA1 and BNP protein decreased,the surface area of cardiomyocyte reduced,the expression of ACSL4 protein decreased,and the expression of SLC7A11 and GPX4 proteins increased(P＜0.05 or P＜0.01).Compared with the ISO group,the LV-CRTC2+ISO group showed a decrease in surface area of cardiomyocytes(P＜0.01),a decrease in ACTA1,BNP and ACSL4 protein ex-pression,an increase in SLC7A11 and GPX4 protein expression,and a decrease in ROS and iron ion content(P＜0.05 or P＜0.01).Conclusion CRTC2 alleviates cardiomyocyte hypertrophy and protect cardiac function by suppressing fer-roptosis in cardiomyocytes.

Aim To explore the role and mechanism of epimedokoreanin B(EKB)in HepaRG cell pyroptosis through endoplasmic reticulum stress and NLRP1-me-diated pyroptosis pathway.Methods The effect of EKB on the viability of HepaRG cells at different con-centrations was determined by MTT assay,and the cell growth status was recorded by Incucyte.Four groups of HepaRG cells were set up.The control group was cul-tured with complete medium for 24 h;the drug admin-istration group was cultured with three concentration gradients of 6.25,12.5 and 25 μmol·L-1 of EKB for 24 h.Western blot was used to detect the expression levels of endoplasmic reticulum stress-related proteins and pyroptosis-related proteins in the cells of each group.Results HepaRG cells showed cytotoxicity at a concentration of 6.25 μmol·L-1 for 24 h,and the half maximal inhibitory concentration(IC50)was 12.41 μmol·L-1.Incucyte recordings of the cell growth status showed that the cells in the control group were in good growth status,and the vesicular pyropto-sis cells appeared in the different concentrations of EKB and the cells swelled and ruptured after 24 h.Western blot showed that the protein expression levels of endoplasmic reticulum stress-related proteins pERK,eIF-2α,ATF-4,GRP78,and CHOP significantly in-creased in HepaRG cells at 25 μmol·L-1 of EKB compared with the control group.The proteins of the classical pathway of cellular pyroptosis mediated by NLRP1,caspase-1,cleaved caspase-1,GSDMD,GS-DMD-N significantly increased in HepaRG cells.Con-clusion EKB administration induces HepaRG cell py-roptosis,and EKB activates HepaRG cells to undergo endoplasmic reticulum stress and activates the NLRP1/caspase-1/GSDMD-mediated pyroptosis pathway.

Aim To investigate the role and regulatory mechanism of CREB regulated transcription coactivator 2(CRTC2)in cardiomyocyte hypertrophy.Methods A pathological cardiomyocyte hypertrophy model was established in C57BL/6 mice by intraperitoneal injection of isoproterenol(ISO),the expression of CRTC2 in cardiac tissue was detec-ted by Western blot.The CRTC2 knockout mice model was constructed,the cardiac function of mice was detected by small animal echocardiography,the collagen fiber content in mice cardiac tissue was detected by Masson staining,the car-diomyocyte hypertrophy related proteins:skeletal muscle α1-actin(ACTA1)and brain natriuretic peptide(BNP),as well as ferroptosis related proteins:acyl-CoA synthetase long chain family member 4(ACSL4),solute carrier family 7 member 11(SLC7A11)and glutathione peroxidase 4(GPX4)in mice cardiac tissue were detected by Western blot,the iron ion content in mice cardiac tissue was detected by iron ion kit,to evaluate the correlation between CRTC2 and cardiomyocyte hypertrophy and ferroptosis.H9c2 cells were induced by ISO to construct an in vitro model of cardiomyocyte hypertrophy,the protein expressions of CRTC2,ACTA1,BNP,ACSL4,SLC7A11 and GPX4 were detected after intervention with fer-roptosis inhibitor ferrostatin-1(Fer-1).H9c2 cells with CRTC2 overexpression induced by ISO were used to construct an in vitro model of cardiomyocyte hypertrophy,the related indicators of cardiomyocyte hypertrophy and ferroptosis were detec-ted to explore the mechanism of CRTC2 in cardiomyocyte hypertrophy.Results Compared with the control group,the expression of CRTC2 protein in the cardiac tissue of ISO induced cardiomyocyte hypertrophy mice was increased(P＜0.05).Compared with wild-type mice,CRTC2-/-mice showed worsened cardiac function,manifested as increased left ventricular end-diastolic diameter(LVEDD),left ventricular end-systolic diameter(LVESD),left ventricular posterior wall thickness(LVPWT),heart weight/tibia length(HW/TL)and heart weight/body weight(HW/BW),decreased short axis shortening(FS)and ejection fraction(EF),increased collagen fiber content in cardiac tissue,upregulated ex-pression of cardiomyocyte hypertrophy-related proteins ACTA1 and BNP,increased mRNA and protein expression of ferrop-tosis-related protein ACSL4,decreased mRNA and protein expression of SLC7A11 and GPX4,and elevated iron ion content in cardiac tissue(P＜0.05 or P＜0.01).In vitro experiments showed that compared with ISO group,the ISO+Fer-1 group had no significant change in CRTC2 protein expression(P＞0.05),the expression of ACTA1 and BNP protein decreased,the surface area of cardiomyocyte reduced,the expression of ACSL4 protein decreased,and the expression of SLC7A11 and GPX4 proteins increased(P＜0.05 or P＜0.01).Compared with the ISO group,the LV-CRTC2+ISO group showed a decrease in surface area of cardiomyocytes(P＜0.01),a decrease in ACTA1,BNP and ACSL4 protein ex-pression,an increase in SLC7A11 and GPX4 protein expression,and a decrease in ROS and iron ion content(P＜0.05 or P＜0.01).Conclusion CRTC2 alleviates cardiomyocyte hypertrophy and protect cardiac function by suppressing fer-roptosis in cardiomyocytes.

Aim To explore the role and mechanism of epimedokoreanin B(EKB)in HepaRG cell pyroptosis through endoplasmic reticulum stress and NLRP1-me-diated pyroptosis pathway.Methods The effect of EKB on the viability of HepaRG cells at different con-centrations was determined by MTT assay,and the cell growth status was recorded by Incucyte.Four groups of HepaRG cells were set up.The control group was cul-tured with complete medium for 24 h;the drug admin-istration group was cultured with three concentration gradients of 6.25,12.5 and 25 μmol·L-1 of EKB for 24 h.Western blot was used to detect the expression levels of endoplasmic reticulum stress-related proteins and pyroptosis-related proteins in the cells of each group.Results HepaRG cells showed cytotoxicity at a concentration of 6.25 μmol·L-1 for 24 h,and the half maximal inhibitory concentration(IC50)was 12.41 μmol·L-1.Incucyte recordings of the cell growth status showed that the cells in the control group were in good growth status,and the vesicular pyropto-sis cells appeared in the different concentrations of EKB and the cells swelled and ruptured after 24 h.Western blot showed that the protein expression levels of endoplasmic reticulum stress-related proteins pERK,eIF-2α,ATF-4,GRP78,and CHOP significantly in-creased in HepaRG cells at 25 μmol·L-1 of EKB compared with the control group.The proteins of the classical pathway of cellular pyroptosis mediated by NLRP1,caspase-1,cleaved caspase-1,GSDMD,GS-DMD-N significantly increased in HepaRG cells.Con-clusion EKB administration induces HepaRG cell py-roptosis,and EKB activates HepaRG cells to undergo endoplasmic reticulum stress and activates the NLRP1/caspase-1/GSDMD-mediated pyroptosis pathway.