Researchers commonly use cyclization recombination enzyme/locus of X-over P1 (Cre/loxP) technology-based conditional gene knockouts of model mice to investigate the functional roles of genes of interest in Sertoli and Leydig cells within the testis. However, the shortcomings of these genetic tools include high costs, lengthy experimental periods, and limited accessibility for researchers. Therefore, exploring alternative gene silencing techniques is of great practical value. In this study, we employed adeno-associated virus (AAV) as a vector for gene silencing in Sertoli and Leydig cells. Our findings demonstrated that AAV serotypes 1, 8, and 9 exhibited high infection efficiency in both types of testis cells. Importantly, we discovered that all three AAV serotypes exhibited exquisite specificity in targeting Sertoli cells via tubular injection while demonstrating remarkable selectivity in targeting Leydig cells via interstitial injection. We achieved cell-specific knockouts of the steroidogenic acute regulatory ( Star ) and luteinizing hormone/human chorionic gonadotropin receptor (Lhcgr) genes in Leydig cells, but not in Sertoli cells, using AAV9-single guide RNA (sgRNA)-mediated gene editing in Rosa26-LSL-Cas9 mice. Knockdown of androgen receptor ( Ar ) gene expression in Sertoli cells of wild-type mice was achieved via tubular injection of AAV9-short hairpin RNA (shRNA)-mediated targeting. Our findings offer technical approaches for investigating gene function in Sertoli and Leydig cells through AAV9-mediated gene silencing.
Animals ; Male ; Leydig Cells/metabolism* ; Mice ; Dependovirus/genetics* ; Sertoli Cells/metabolism* ; Gene Silencing ; Genetic Vectors ; Testis/cytology*

Objective To compare the effectiveness between ultrasound-guided paravertebral block(PVB)and general anesthesia in percutaneous nephrolithotomy(PCNL).Methods Clinical data of 284 patients who underwent PCNL in our hospital from January 2023 to March 2024 were collected.These were 78 cases of PVB(PVB group)and 206 cases of general anesthesia(general anesthesia group).A total of 61 cases in each of the two groups were matched with propensity score,and the operative time,intraoperative fluid infusion volume,postoperative haemoglobin changes,postoperative visual analogue scale(VAS),hospital stay,hospital costs,and incidence of postoperative complications were compared.Results The operations were completed in both groups of patients successfully,with stable vital signs during the operation and no intraoperative anesthetic complications such as bradycardia,respiratory depression,or hypotension.There were significant differences between the two groups in heart rate(HR)before the start of surgery[(66.5±10.4)beats/min vs.(77.5±11.7)beats/min,t=-5.471,P=0.000],HR at the end of surgery[(60.5±8.4)beats/min vs.(71.0±12.3)beats/min,t=-5.472,P=0.000],mean arterial pressure(MAP)before the start of surgery[(97.2±11.9)mm Hg vs.(103.1±12.3)mm Hg,t=-2.694,P=0.008],MAP at the end of surgery[(85.2±8.8)mm Hg vs.(94.3±11.6)mm Hg,t=-4.892,P=0.000],pulse oxygen saturation(SpO2)before the start of surgery[(99.0±1.1)％vs.(99.6±0.6)％,t=-3.347,P=0.001]and SpO2 at the end of surgery[(99.2±1.1)％vs.(99.8±0.4)％,t=-4.122,P=0.000].The operative time was significantly shorter in the PVB group than that in the general anesthesia group[55.0(41.5,75.5)min vs.95.0(65.0,130.0)min,Z=-5.173,P=0.000].The postoperative haemoglobin changes[5(2,11)g/L vs.8(4,11)g/L,Z=-2.099,P=0.036],postoperative VAS score[1(1,1)points vs.1(1,2)points,Z=-3.342,P=0.001],postoperative hospital stay[3(2,3)dvs.4(3,6)d,Z=-6.016,P=0.000]and hospital costs[(14 499.4±2141.0)yuan vs.(19 634.2±3846.8)yuan,t=-9.109,P=0.000]were statistically lower in the PVB group than those in the general anesthesia group.Comparison of intraoperative fluid infusion volume[600(600,600)ml vs.600(600,1100)ml,Z=-1.800,P=0.072],postoperative venting time[1(1,1)d vs.1(1,1)d,Z=-1.045,P=0.296],and phase Ⅰ stone removal rate[88.5％(54/61)vs.82.0％(50/61),x2=1.043,P=0.307]showed no significant differences between the two groups.The incidence of postoperative complications in the PVB group was significantly lower than that in the general anesthesia group[6.6％(4/61)vs.19.7％(12/61),x2=4.604,P=0.032].Conclusion Ultrasound-guided PVB in PCNL maintains stable intraoperative vital signs,reduces operative time and bleeding,alleviates postoperative pain,reduces postoperative complications and postoperative hospital stay,and saves medical costs,having a good safety and efficacy.

Objective:To compare the differences of three-dimensional computed tomography(3DCT),four-dimensional computed tomography(4DCT),and multi-parametric magnetic resonance(MR)sequences of the radiotherapy for liver cancer in delineation for target area,and analyze which MR sequence was more accurate in assisting CT image to delineate the target area,and design respectively reverse intensity modulated radiotherapy plan,and compare the dosimetric parameters of the target areas of receiving radiotherapy and normal liver tissue.Methods:This retrospective study was conducted to analyze radiotherapy data from case data of 18 patients with hepatocellular carcinoma(HCC)who admitted to the First Affiliated Hospital of Bengbu Medical University between August 2023 and June 2024.These data included 10 respiratory phases in 3DCT and 4DCT,and free-breathing sequence(MR-FB),diaphragm navigation sequence(MR-NAVI),and breath-hold(MR-BH)sequence of multi-parametric MRI,and the gross tumor volumes(GTVs)of them were delineated,which were respectively 5 modal images and liver contours included GTV3D,GTV4D,GTVMR-FB,GTVMR-NAVI,and GTVMR-BH.Pinnacle3 9.8 treatment plan system(TPS)was applied to conduct registration and fusion for image by using mutual information method.The 3DCT(GTV3D)was used as reference image to compare the volume of target area,and the displacement of mass center with other 4 kinds of images.The volume difference(VD),overlap ratio(OR),Dice similarity coefficient(DSC),and Jaccard index(JAC)were used to assess the differences among different target area.The indicators of plan evaluation included conformity index(CI),homogeneity index(HI),GTV doses(D2%,D98%,Dmean),and the exposure dose of normal tissue of liver.Results:In images of five modalities,the GTV median volumes were respectively 28.83,33.10,26.75,25.05,and 22.65 cm3.In images of five modalities,the median volume of liver were respectively 1293.46,1483.09,1213.81,1195.69,and 1141.02 cm3.Compared with other 3 target areas,the displacement of GTVMR-BH was the smallest on head-foot direction,with statistically significant differences among them(Z=-2.305,-2.307,-2.134,P<0.05).The OR,DSC,and JAC values of GTV4D were significantly better than these of GTVMR-FB,GTVMR-NAVI,and GTVMR-BH(ZOR=-2.911,-3.006,-3.195,ZDSC=-2.726,-2.215,-2.556,ZJAC=-2.556,-2.704,-2.953,P<0.05).The VD value of GTVMR-FB was better than that of GTV4D,GTVMR-NAVI,and GTVMR-BH,with statistically significant(Z=-2.675,-2.817,-2.580,P<0.05).Additionally,the OR,DSC,and JAC values of GTVMR-FB and GTVMR-NAVI were better than those of GTVMR-BH,with statistically significant(ZOR=-2.859,-2.817,ZDSC=-2.184,-2.783,ZJAC=-2.385,-2.783,P<0.05).All five plans met clinical dose requirements.Friedman test showed there was no statistically significant differences in dosimetric parameters of target area among different plans(P>0.05).However,compared to the PTV3D plan,the PTVMR-FB,PTVMR-NAVI,and PTVMR-BH plans resulted in lower levels in mean dose(Dmean)of liver and volume parameters(V5,V10,V20,V30)of various doses,with statistically significant differences(Dmean:Z=-2.433,-2.307,-2.807,ZV5=-2.512,-2.433,-2.652,ZV10=-2.433,-2.536,-2.968,ZV20=-2.536,-2.652,-2.807,ZV30=-2.611,-2.652,-2.968,P<0.05).Conclusion:In actually clinical application,MR-NAVI and 4DCT also can be adopted to assist 3DCT to delineate target area besides MR-FB sequence that is conventionally used in MR location,thus can enhance precision of delineation,and optimize radiotherapy plan,and decrease exposure dose of normal liver tissue.

Objective:To investigate the correlation between frailty and immune markers in elderly patients diagnosed with heart failure with preserved ejection fraction(HFpEF).Methods:A total of 416 elderly patients with HFpEF, who were hospitalized in the Department of Geriatrics at Henan Provincial People's Hospital from March 2021 to December 2023, were selected as research subjects.The Fried frailty phenotype was employed to assess frailty.Fasting venous blood samples were collected to measure levels of CD4+ T lymphocytes, CD8+ T lymphocytes, the CD4+ /CD8+ ratio, and immunoglobulins A, M, and G. Spearman correlation analysis and multiple linear regression analysis were conducted to evaluate the relationship between frailty scores and immune markers.Results:Spearman correlation analysis revealed a significant association between frailty score and the CD4+ /CD8+ ratio( r=-0.659, P<0.001), immunoglobulin A( r=-0.454, P<0.001), immunoglobulin M( r=-0.522, P<0.001), and immunoglobulin G( r=-0.802, P<0.001).Furthermore, multiple linear regression analysis indicated that, after adjusting for confounding factors, frailty score served as a significant negative predictor of the CD4+ /CD8+ ratio( β=-0.562, P<0.001), immunoglobulin A( β=-0.366, P<0.001), immunoglobulin M( β=-0.445, P<0.001), and immunoglobulin G( β=-0.772, P<0.001).In comparison to the non-frail group, the frail group exhibited significantly lower values for the CD4+ /CD8+ ratio( β=-0.666, P<0.001)and levels of immunoglobulin A( β=-0.514, P<0.001), immunoglobulin M( β=-0.526, P<0.001), and immunoglobulin G( β=-0.814, P<0.001). Conclusions:In hospitalized elderly patients with heart failure with HFpEF, frailty serves as an independent risk factor for the reduction of the CD4+ /CD8+ ratio, as well as levels of immunoglobulin A, immunoglobulin M, and immunoglobulin G. Furthermore, the frailty score demonstrates a significant negative predictive value for these immunological markers.Therefore, it is essential to enhance our understanding of frailty and to prioritize its prevention and treatment, as this may help mitigate immune dysfunction and promote recovery in elderly patients.

Objective:To evaluate the value of multi-slice spiral CT (MSCT) in the diagnosis of liver metastases with rich blood supply.Methods:The clinical data and imaging data of 19 patients with liver metastases with rich blood supply admitted to the 921st Hospital of the Joint Logistics Support Force of Chinese People′s Liberation Army from September 2018 to September 2023 were retrospectively analyzed, and the number, location, shape, size of the lesions and the images of CT plain scan and dynamic enhanced scan were analyzed.Results:Among the 19 patients, there were 18 multiple cases and 1 single case. A total of 108 lesions were found. There were 62 cases (57.4%) in the right lobe of liver and 87 cases (80.6%) in the peripheral part of liver. The form of circular or quasi-circular, there were 99, irregular shape or lobed 9. The focal diameter was 0.6-6.8 cm. CT plain scan showed that 99 lesions showed slightly low density, and the other 9 lesions showed equal density relative to the background liver. In the dynamic enhanced scan, 108 lesions in arterial stage showed high-density enhancement, 97 lesions showed circular enhancement, and 11 lesions showed nodular enhancement. Among them, 77 lesions had moderate to obvious intensification density. Of the 108 lesions in the portal vein stage, 31 lesions showed moderate to obvious enhancement density, 49 lesions showed slightly low clearance density, and 28 lesions showed continuous enhancement density. In the delayed stage, all 108 lesions showed slightly low density.Conclusions:The main features of liver metastases with rich blood supply are: low density on plain CT scan, annular or nodular enhancement in the arterial phase of enhanced CT scan, and the peak of enhanced density can be in the arterial phase or the portal vein phase. Combined with clinical data, CT can make a correct diagnosis.

Objective:To investigate the effects of short-term olfactory training and visual stimulation on olfactory recognition function and quality of life in patients following functional endoscopic sinus surgery (FESS).Methods:A total of 80 patients who underwent FESS in the Department of Otorhinolaryngology Head and Neck Surgery at the Affiliated Hospital of Xuzhou Medical University between December 2023 and February 2024 were enrolled in this study. The cohort comprised 67 males and 13 females, aged from 17 to 75 years. Participants were randomly allocated to either a control group ( n=40) or an experimental group ( n=40). The participants in control group received routine postoperative management, including nasal irrigation, oral antibiotics and glucocorticoids, topical budesonide nasal spray, and hypertonic saline solution for 12 weeks. In addition to the standard care, the participants in experimental group underwent olfactory-visual stimulation training starting 24 hours postoperatively, lasting for 2 weeks. The olfactory identification test (OIT), visual analogue scale (VAS) for olfaction and Questionnaire of Olfactory Disorders of Life (QOD-QoL, hereafter referred to as QOD) were administered preoperatively. Follow-up assessments were performed 2 weeks and 3 months postoperatively, with the same tests repeated. Data were analyzed using SPSS 27.0 and GraphPad Prism 7 statistical software. Results:There was no significant difference in preoperative OIT, VAS, and QOD scores between the two groups. The olfactory recognition function of some patients was improved after removing the packing material 24 hours after FESS surgery. The OIT scores of 2 weeks post-surgery were significantly higher than preoperative values in both groups (the experimental group Z=-4.73, P<0.001; the control group Z=-4.73, P<0.001). Participants in both groups showed improvements in olfactory VAS and QOD scores (experimental group Z value was -2.88 and -5.45, P<0.01 and<0.001, respectively; the control group Z value was -4.42 and -5.50, respectively, both P<0.001). However, there was no significant difference in the VAS score between the two groups ( Z=-0.68, P=0.499). The paticipants in experimental group showed greater improvement in OIT and greater reduction in QOD scores compared to the control group ( Z=-2.19, P=0.029; Z=-2.99, P=0.003). There was no significant difference in the decrease of olfactory VAS between the two groups ( Z=-0.02, P=0.988). There were no statistically significant differences of all patients in VAS, OIT and QOD scores at 2 weeks and 3 months after surgery (experimental group Z value was -0.91, -0.90 and -1.43, respectively, all P>0.05; control group Z value was -1.21, -0.84 and -0.91, respectively, all P>0.05). At 3 months post-surgery, the OIT scores in the experimental group were higher, the QOD scores were lower than those in the control group ( Z=-2.89, P=0.004; Z=-2.87, P=0.004). Conclusion:Short-term olfactory-visual stimulation training in the early postoperative period of FESS significantly improves the olfactory recognition function and enhances the quality of life of patients.

Objective To explore the prediction efficiency of Ki-67 expression status in pituitary adenoma(PA)based on a joint model of siamese network and transfer learning.Methods The preoperative MR T1WI enhanced sequence images of 370 patients with PA diagnosed by surgery and pathology were retrospectively collected.According to the results of immunohistochemical,all patients were divided into high proliferation index group(Ki-67≥3,n=97)and low proliferation index group(Ki-67＜3,n=273),and all the sample data were randomly divided into training set and test set according to the ratio of 7∶3.Two single predictive models,ResNet-50 and VGGNet-16 and combined them with the siamese network and transfer learning were built.The prediction efficiency of different models were evaluated via accuracy rate,precision rate,recall rate,F1 score and receiver operating characteristic(ROC)curve as the main criteria.Results Compared with a single predictive model,the model combined with siamese network and transfer learning showed a better performance for predicting the Ki-67 of PA.Additionally,the ResNet-50 joint model exhibited the superior predictive performance.The accuracy rate was 0.872 7,the precision rate was 0.812 5,the recall rate was 0.764 7,the F1 score was 0.787 9,and the area under the curve(AUC)was 0.841 6.Conclusion The joint model based on siamese network and transfer learning exhibits a higher efficiency for predicting the Ki-67 expression status in PA,which can help the clinicians to formulate more personalized treatment for the patients.

Objective To investigate the effects of human milk on serum bilirubin levels and gut microbiota in neonatal rats with hyperbilirubinemia.Methods A total of 24 7-day-old specific-pathogen-free Sprague-Dawley rats were injected with bilirubin or normal saline,respectively,and human milk or formula milk was administered 24 hours later for intervention.The rats were divided randomly into four groups:human milk-normal saline group(HN),human milk-bilirubin group(HB),formula milk-normal saline group(FN),and formula milk-bilirubin group(FB).Samples were taken 72 hours later,and serum bilirubin values were detected by enzyme-linked immunosorbent assay.The intestinal microbiota were analyzed using 16S rDNA high-throughput sequencing.Results There was no significant difference in bilirubin values among the groups.Pseudomonas was negatively correlated with indirect bilirubin value(P＜0.05).The composition of the intestinal microbiota differed significantly between human milk and formula milk after gastric administration,with Firmicutes(P＜0.01),Enterococci(P＜0.05),being the main microbiota in the HN and HB groups,and Proteobacteria(P＜0.001),Escherichia Shigella(P＜0.01)and Acinetobacter(P＜0.01)being the main in the FN and FB groups.Conclusions Pseudomonas may be negatively associated with bilirubin,and the structure of the intestinal microbiota may differ in relation to human milk and formula feeding.

Tumor drug resistance is an important problem in the failure of chemotherapy and targeted drug therapy, which is a complex process involving chromatin remodeling. SWI/SNF is one of the most studied ATP-dependent chromatin remodeling complexes in tumorigenesis, which plays an important role in the coordination of chromatin structural stability, gene expression, and post-translation modification. However, its mechanism in tumor drug resistance has not been systematically combed. SWI/SNF can be divided into 3 types according to its subunit composition: BAF, PBAF, and ncBAF. These 3 subtypes all contain two mutually exclusive ATPase catalytic subunits (SMARCA2 or SMARCA4), core subunits (SMARCC1 and SMARCD1), and regulatory subunits (ARID1A, PBRM1, and ACTB, etc.), which can control gene expression by regulating chromatin structure. The change of SWI/SNF complex subunits is one of the important factors of tumor drug resistance and progress. SMARCA4 and ARID1A are the most widely studied subunits in tumor drug resistance. Low expression of SMARCA4 can lead to the deletion of the transcription inhibitor of the BCL2L1 gene in mantle cell lymphoma, which will result in transcription up-regulation and significant resistance to the combination therapy of ibrutinib and venetoclax. Low expression of SMARCA4 and high expression of SMARCA2 can activate the FGFR1-pERK1/2 signaling pathway in ovarian high-grade serous carcinoma cells, which induces the overexpression of anti-apoptosis gene BCL2 and results in carboplatin resistance. SMARCA4 deletion can up-regulate epithelial-mesenchymal transition (EMT) by activating YAP1 gene expression in triple-negative breast cancer. It can also reduce the expression of Ca²⁺ channel IP3R3 in ovarian and lung cancer, resulting in the transfer of Ca²⁺ needed to induce apoptosis from endoplasmic reticulum to mitochondria damage. Thus, these two tumors are resistant to cisplatin. It has been found that verteporfin can overcome the drug resistance induced by SMARCA4 deletion. However, this inhibitor has not been applied in clinical practice. Therefore, it is a promising research direction to develop SWI/SNF ATPase targeted drugs with high oral bioavailability to treat patients with tumor resistance induced by low expression or deletion of SMARCA4. ARID1A deletion can activate the expression of ANXA1 protein in HER2+ breast cancer cells or down-regulate the expression of progesterone receptor B protein in endometrial cancer cells. The drug resistance of these two tumor cells to trastuzumab or progesterone is induced by activating AKT pathway. ARID1A deletion in ovarian cancer can increase the expression of MRP2 protein and make it resistant to carboplatin and paclitaxel. ARID1A deletion also can up-regulate the phosphorylation levels of EGFR, ErbB2, and RAF1 oncogene proteins.The ErbB and VEGF pathway are activated and EMT is increased. As a result, lung adenocarcinoma is resistant to epidermal growth factor receptor tyrosine kinase inhibitors (EGFR-TKIs). Although great progress has been made in the research on the mechanism of SWI/SNF complex inducing tumor drug resistance, most of the research is still at the protein level. It is necessary to comprehensively and deeply explore the detailed mechanism of drug resistance from gene, transcription, protein, and metabolite levels by using multi-omics techniques, which can provide sufficient theoretical basis for the diagnosis and treatment of poor tumor prognosis caused by mutation or abnormal expression of SWI/SNF subunits in clinical practice.

Background High-sensitivity C-reactive protein (hs-CRP) is a sensitive biomarker for cardiovascular disease (CVD) and can independently predict the risk of cardiovascular events. Although the association between per- and polyfluoroalkyl substances (PFAS) exposure and CVD risk has been widely reported, studies on the association between hs-CRP and PFAS remain limited. Objective To investigate the association between PFAS and hs-CRP levels, to provide a scientific basis for early identification and prevention of environment-related cardiovascular events. Methods This study utilized data from the National Health and Nutrition Examination Survey (NHANES) database (2015–2018). Based on predefined inclusion and exclusion criteria, a total of 3219 participants were included for subsequent analysis. Five PFAS with detection rates exceeding 90% were selected as exposure factors and log-transformed (lnPFAS) for subsequent analysis, including perfluorohexane sulfonic acid (PFHxS), perfluorononanoic acid (PFNA), perfluorooctanoic acid (PFOA), perfluorooctane sulfonic acid (PFOS), and perfluorodecanoic acid (PFDA). After selecting potential confounders through a change-in-estimate approach, logistic regression was employed to examine the effects of individual PFAS on serum hs-CRP level. Additionally, quantile g-computation and Bayesian kernel machine regression (BKMR) were applied to assess the combined effects of PFAS mixtures on hs-CRP. Results The study included 3219 participants with a mean age of (50.4±17.6) years, of whom 1554 (48.28%) were male. The median concentrations of the five PFAS compounds were: 1.20 ng·mL⁻¹ (PFHxS), 0.50 ng·mL⁻¹ (PFNA), 1.50 ng·mL⁻¹ (PFOA), 5.20 ng·mL⁻¹ (PFOS), and 0.20 ng·mL⁻¹ (PFDA). The Wilcoxon rank-sum tests revealed statistically significant differences between the elevated serum hs-CRP (≥2 mg·L⁻¹) and normal (<2 mg·L⁻¹) groups for all five PFAS compounds (PFHxS: P=0.005; PFNA: P=0.009; PFOA: P<0.001; PFOS: P<0.001; PFDA: P<0.001). After adjusting for potential confounders, the analysis of individual PFAS exposure demonstrated significant negative associations between serum hs-CRP levels and both ln-PFOS (OR: 0.87; 95%CI: 0.79, 0.96; P=0.0036) and lnPFDA (OR: 0.78; 95%CI: 0.71, 0.86; P < 0.001). The joint effect analysis using quantile g-computation revealed an overall inverse association between PFAS mixtures and hs-CRP, with lnPFDA emerging as a primary contributor; an similar association was observed in males, but not significant in females. Further evaluation through BKMR showed that when PFAS mixture concentrations reached or exceeded P₇₀, their combined effect on hs-CRP became significantly lower than when all PFAS concentrations were at P₅₀, establishing a clear negative dose-response relationship. Conclusion The mixed exposure to PFAS showed a significant negative joint effect on serum hs-CRP levels, with PFDA being the key component contributing to this combined effect. Future prospective cohort studies are warranted to further validate these findings and elucidate the underlying mechanisms involved.