OBJECTIVE To investigate the protective effect and mechanism of cryptotanshinone （CTS） on heart and kidney function in rats with cardiorenal syndrome （CRS） by regulating phosphoinositide kinase-3 （PI3K）/protein kinase B （Akt）/ mammalian target of rapamycin （mTOR） signaling pathway. METHODS CRS model of rats was induced by left anterior descending coronary artery ligation combined with acute renal ischemia-reperfusion injury. Model rats were randomly divided into CRS model group （CRS group）， low-dose CTS group （CTS-L）， high-dose CTS group （CTS-H group）， high-dose CTS+PI3K activator 740Y-P group （CTS-H+740Y-P group）， with 12 rats in each group. Another 12 rats were selected as the normal control group （Normal group） and were carried out surgery without modeling. CTS-L group and CTS-H group were respectively given CTS 30 and 60 mg/kg intragastrically， once a day， for consecutive 14 d. Besides the intervention of CTS 60 mg/kg intragastrically， CTS-H+740Y-P group was given 10 mg/kg 740Y-P intraperitoneally， once a day， for 14 consecutive days. After the last medication， the levels of cardiac function [left ventricular ejection fraction （LVEF）， left ventricular end-systolic diameter （LVESD）， left ventricular end-diastolic diameter （LVEDD）， left ventricular fraction shortening （LVFS）] and renal function [24 h urinary protein， blood urea nitrogen （BUN）， serum creatinine （Scr）， brain natriuretic peptide （BNP）] were detected in rats. The pathological changes and fibrosis of the heart and kidney in rats were observed； the expressions of PI3K/Akt/mTOR signaling pathway in heart and renal tissue were all detected. RESULTS Compared with Normal group， the levels of LVEF and LVFS in rats were all decreased significantly in CRS group （P＜0.05）； the levels of LVESD， LVEDD， 24 h urinary protein， serum levels of BUN， Scr and BNP， collagen area and the phosphorylation of PI3K， Akt and mTOR protein in heart and renal tissue were all increased significantly （P＜0.05）. The morphology of myocardial cells was enlarged and disordered； the structure ofrenal tubules was disordered， epithelial cells were wrinkled， and there was infiltration of inflammatory cells. Compared with CRS group， the above indexes of rats were reversed significantly in CTS-L group and CTS-H group （P＜0.05）； heart and kidney function had been restored， and pathological damage and fibrosis had been reduced. PI3K activator 740Y-P weakened the protective effect of CTS on cardiac and renal function in CRS rats. CONCLUSIONS CTS can protect heart and kidney function in CRS rats， the mechanism of which may be associated with inhibiting the PI3K/Akt/mTOR signaling pathway.

Objective The critical genes associated with exhausted CD8+T cells were screened and validated by mapping the single-cell transcriptome profile of high-grade serous ovarian cancer(HGSOC).Methods The specific subtypes of T cells in the tumor microenvironment were analyzed using the single-cell sequencing data from the early stage of laboratory(SRA database:PRJNA756768)and integrating 5 HGSOC sequencing from the database,and the differentiation trajectory of T cell subsets was ex-plored through pseudotime analysis.Differential gene enrichment was used to determine immunosuppressed CD8+IL-2Low and CD8+IFN-γLow T cell subsets and differential genes,and candidate molecules closely related to exhausted CD8+T cells were screened based on patient prognosis.Flow cytometry was used to analyze the expression of PHLDA1 on CD8+T cells,CD4+T cells and Treg cells dur-ing the activation to exhaustion process of T cells in human PBMCs.ELISA was used to detect the levels of IFN-γ and IL-2 secreted by CD8+T cells in PHLDA1High and PHLDA1Low.Finally,flow cytometry was used to analyze the association between PHLDA1 and ex-hausted markers PD-1 and TIM-3.Results The results showed that T cells were grouped in three ways:(1)IL-2High and IL-2Low;(2)IFN-γHigh and IFN-γLow;and(3)exhausted and cytotoxic CD8+T cells.Subsequently,the intersection of its differentially expressed genes was taken,and the key gene PHLDA1 was ultimately screened.Flow cytometry analysis suggested that during the process of T cell activation to exhaustion,the expression of PHLDA1 continued to increase on CD8+T cells,CD4+T cells and Treg cells;The ELISA results showed that the levels of IFN-γ and IL-2 secreted by CD8+PHLDA1High T cells were significantly lower than those of CD8+PHLDA1Low T cells.Meanwhile,the CD8+PHLDA1High T cell subset could simultaneously cover the exhausted T cell types of CD8+TIM-3+and CD8+PD-1+.Conclusion Based on single-cell sequencing data,this study identified PHLDA1 as a key molecule responsi-ble for CD8+T cell exhaustion in OC,providing new insights for immunotherapy of OC.

Ferroptosis (FPT), a novel form of programmed cell death, is characterized by overwhelming iron/reactive oxygen species (ROS)-dependent accumulation of lipid peroxidation (LPO). However, the insufficiency of endogenous iron and ROS level limited the FPT therapeutic efficacy to a large extent. To overcome this obstacle, the bromodomain-containing protein 4 (BRD₄)-inhibitor (+)-JQ1 (JQ1) and iron-supplement ferric ammonium citrate (FAC)-loaded gold nanorods (GNRs) are encapsulated into the zeolitic imidazolate framework-8 (ZIF-8) to form matchbox-like GNRs@JF/ZIF-8 for the amplified FPT therapy. The existence of matchbox (ZIF-8) is stable in physiologically neutral conditions but degradable in acidic environment, which could prevent the loaded agents from prematurely reacting. Moreover, GNRs as the drug-carriers induce the photothermal therapy (PTT) effect under the irradiation of near-infrared II (NIR-II) light owing to the absorption by localized surface plasmon resonance (LSPR), while the hyperthermia also boosts the JQ1 and FAC releasing in the tumor microenvironment (TME). On one hand, the FAC-induced Fenton/Fenton-like reactions in TME can simultaneously generate iron (Fe³⁺/Fe²⁺) and ROS to initiate the FPT treatment by LPO elevation. On the other hand, JQ1 as a small molecule inhibitor of BRD₄ protein can amplify FPT through downregulating the expression of glutathione peroxidase 4 (GPX4), thus inhibiting the ROS elimination and leading to the LPO accumulation. Both in vitro and in vivo studies reveal that this pH-sensitive nano-matchbox achieves obvious suppression of tumor growth with good biosafety and biocompatibility. As a result, our study points out a PTT combined iron-based/BRD₄-downregulated strategy for amplified ferrotherapy which also opens the door of future exploitation of ferrotherapy systems.

Liver fibrosis is a reversible pathological process caused by chronic liver damage and a major risk factor for hepatocellular carcinoma (HCC). Hepatic stellate cell (HSC) activation is considered the main target for liver fibrosis therapy. However, the efficiency of this strategy is limited due to the complex microenvironment of liver fibrosis, including excessive extracellular matrix (ECM) deposition and hypoxia-induced imbalanced ECM metabolism. Herein, nilotinib (NIL)-loaded hyaluronic acid (HA)-coated Ag@Pt nanotriangular nanozymes (APNH NTs) were developed to inhibit HSCs activation and remodel the microenvironment of liver fibrosis. APNH NTs efficiently eliminated intrahepatic reactive oxygen species (ROS) due to their inherent superoxide dismutase (SOD) and catalase (CAT) activities, thereby downregulating the expression of NADPH oxidase-4 (NOX-4) and inhibiting HSCs activation. Simultaneously, the oxygen produced by the APNH NTs further alleviated the hypoxic microenvironment. Importantly, the released NIL promoted collagen depletion by suppressing the expression of tissue inhibitor of metalloproteinase-1 (TIMP-1), thus synergistically remodeling the microenvironment of liver fibrosis. Notably, an in vivo study in CCl₄-induced mice revealed that APNH NTs exhibited significant antifibrogenic effects without obvious long-term toxicity. Taken together, the data from this work suggest that treatment with the synthesized APNH NTs provides an enlightening strategy for remodeling the microenvironment of liver fibrosis with boosted antifibrogenic activity.

Objective The aim of this study was to draw single-cell transcriptome profiles of high-grade serous ovarian cancer(HGSOC),borderline ovarian cancer(OC),and normal ovaries in order to identify biomarkers that can diagnose and predict the prognosis of OC.Methods The differentially expressed genes between HGSOC,borderline OC,and normal ovarian tissues were ana-lyzed using single-cell data sequenced(SRA database:PRJNA756768).The cell subsets associated with tumor progression were screened by functional enrichment,cell communication between different subsets was analyzed by Cellchat,and cell differentiation traj-ectories were explored by pseudotime analysis to finally determine the subsets most relevant to tumor progression.Combined with OC transcriptome data of OC from the Cancer Genome Atlas(TCGA)with patient prognosis,biomarkers for diagnosing and predicting sur-vival of OC patients were ultimately screened.Results After using t-distribution stochastic neighbor embedding(t-SNE)for di-mensionality reduction,nine cell subpopulations were obtained:endothelial cells,myeloid cells,fibroblasts,T cells,stromal cells,B cells,and 3 epithelial cell subpopulations(C1,C4,and C7).Further analysis revealed that copy number variation(CNV)in the C4 group had the highest score in HGSOC,higher than those of borderline OC and normal ovaries,and was negatively correlated with prognosis.DMKN was a key marker gene in this group.Transcriptome analysis of OC in the TCGA database showed a close correlation between DMKN and poor prognosis(P=0.026),and the diagnostic efficacy of DMKN for OC was significant(A UC=0.906).Con-clusion This study is based on single-cell sequencing data to screen for DMKN,which can effectively diagnose and predict the prognosis of OC.This study provides new ideas for the diagnosis and prognosis prediction of OC.

Objective:To explore the ideal method of minimally invasive anterior lumbar extraperitoneal approach.Methods:Twenty-one adult embalmed cadavers underwent longitudinal incision near the left rectus abdominis, the extraperitoneal space and peritoneal characteristics were observed; the L 2-S 1 disc was exposed through extraperitoneal approach, and the relationship between the anterior large vessels and the disc was observed. One hundred adult abdominal CT were collected to measure the distance between the extraperitoneal fat of anterior abdominal wall and the rectus abdominis and the anterior midline at L 2-S 1 segment. One hundred and fifty adult lumbar MRI were collected to measure the distance between the anterior great vessels and the anterior midline of the intervertebral disc. Fifty-six cases of lumbar fusion were performed by minimally invasive anterior lumbar extraperitoneal approach, including 25 males and 31 females, aged 29-71 years. L 2-L 4 in 8 cases was performed by left rectus abdominis oblique incision, and L 4-S 1 in 48 cases was performed by median left transverse incision, with a length of about 8 cm, the complications related to the surgical approach were evaluated. Results:L 2-L 4 was proximal to the arcuate line, the posterior sheath of rectus abdominis adhered to the peritoneum, which was easy to rupture when separated; the peritoneum gradually thickened from the outer edge of the sheath of rectus abdominis and extraperitoneal fat appears. L 4-S 1 could be exposed distal to the arcuate line, the posterior side of rectus abdominis was extraperitoneal fat, the extension of arcuate line to the lateral abdominal wall would be slightly separated proximally, and there were multiple iliopsoas veins in the medial side of psoas major muscle. L 5S 1 was between the right common iliac artery and the left common iliac vein far, the median sacral vessel was small or absent, and the sympathetic nerve was to the left. Extraperitoneal fat appeared 36.2±9.9 mm, 35.2±11.6 mm and 27.6±11.2 mm away from the outer edge of rectus abdominis at L 2, 3, L 3, 4 and L 4, 5 segments respectively, and covered the posterior side of rectus abdominis and reached the midline at L 5S 1 segment. The left edge of abdominal aorta was 14.9±5.1 mm, 13.9±4.6 mm and 19.7±5.9 mm away from the midline at L 2, 3, L 3, 4 and L 4, 5 level respectively; the inferior vena cava was located on the right side of the midline at L 2, 3 and L 3, 4 level, crossed the midline 4.6±8.7 mm at L 4, 5 level. At L 5S 1 level, the left common iliac vein and the right common iliac artery were 14.6±6.8 mm and 17.6±5.3 mm away from the midline respectively. Seventy-six patients were successfully and fully exposed by small incision through extraperitoneal approach. 1 case of L 4, 5 had iliac lumbar vein tear and hemostasis with bipolar electrocoagulation. The operation time was 70-120 min, with an average of 90 min; Intraoperative bleeding was 15-70 ml, with an average of 30 ml. No severe complication such as nerve and great vessel injury occurred. Conclusion:Minimally invasive lumbar anterior retroperitoneal approach has small trauma and sufficient exposure with good feasibility. L 2-L 4 can be exposed with supine position and oblique incision next to the left rectus abdominis muscle, and L 4~S 1 with French position and median left transverse incision.

Background::Pancreatic adenocarcinoma (PAAD) is an extremely lethal malignancy. Identification of the functional genes and genetic variants related to PAAD prognosis is important and challenging. Previously identified prognostic genes from several expression profile analyses were inconsistent. The regulatory genetic variants that affect PAAD prognosis were largely unknown.Methods::Firstly, a meta-analysis was performed with seven published datasets to systematically explore the candidate prognostic genes for PAAD. Next, to identify the regulatory variants for those candidate genes, expression quantitative trait loci analysis was implemented with PAAD data resources from The Cancer Genome Atlas. Then, a two-stage association study in a total of 893 PAAD patients was conducted to interrogate the regulatory variants and find the prognostic locus. Finally, a series of biochemical experiments and phenotype assays were carried out to demonstrate the biological function of variation and genes in PAAD progression process.Results::A total of 128 genes were identified associated with the PAAD prognosis in the meta-analysis. Fourteen regulatory loci in 12 of the 128 genes were discovered, among which, only rs4887783, the functional variant in the promoter of Ring Finger and WD Repeat Domain 3 ( RFWD3), presented significant association with PAAD prognosis in both stages of the population study. Dual-luciferase reporter and electrophoretic mobility shift assays demonstrated that rs4887783-G allele, which predicts the worse prognosis, enhanced the binding of transcript factor REST, thus elevating RFWD3 expression. Further phenotypic assays revealed that excess expression of RFWD3 promoted tumor cell migration without affecting their proliferation rate. RFWD3 was highly expressed in PAAD and might orchestrate the genes in the DNA repair process. Conclusions::RFWD3 and its regulatory variant are novel genetic factors for PAAD prognosis.

Objective:To explore the application effect of Internet plus group health management in patients with hypertension.Methods:A total of 350 patients with hypertension who were diagnosed for the first time in Beijing Tiantan Hospital, Capital Medical University from May 2020 to May 2021 were selected as the research objects by the convenient sampling method. The random number table method was used to divide the patients into the control group and the intervention group, with 175 cases in each group. Patients in the control group received routine health management, while patients in the intervention group received Internet plus group health management based on the control group. The blood pressure of the two groups was compared before and after intervention.Results:After 6 months of intervention, a total of 15 patients were lost to follow-up. Finally, 165 patients were included in the control group and 170 patients were included in the intervention group. Repeated measurement analysis of variance showed that there were inter-group effects, time effects and interaction effects between groups and time on the variation coefficients of systolic blood pressure, diastolic blood pressure and 24 h systolic blood pressure ( P<0.01) . Conclusions:Internet plus group health management can improve blood pressure control of hypertension patients.

Objective: To evaluate the clinical efficacy of percutaneous tibial nerve electrical stimulation in the treatment of chronic pelvic pain syndrome. Methods: The clinical data of 28 patients with chronic pelvic pain syndrome from January to November 2018 in Dalian Third People′s Hospital were retrospectively analyzed. The patients were treated with percutaneous tibial nerve stimulation. The number of urination in 24 h, number of nocturnal urination, urine volume per urination, quality of life (QOL) score and digital pain intensity score before treatment and after treatment were compared; the patients were followed up for 3 months, and the recurrence and adverse reaction were observed. Results: Compared with those before treatment, the number of urination in 24 h and number of nocturnal urination after treatment were significantly lower: (11.6 ± 6.4) times vs. (20.6 ± 7.8) times and (2.5 ± 1.2) times vs. (5.2 ± 2.6) times, and the urine volume per urination increased significantly: (181.2 ± 65.6) ml vs. (125.4 ± 58.2) ml, the QOL score and digital pain intensity score decreased significantly: (2.6 ± 1.4) scores vs. (5.1 ± 0.8) scores and (2.9 ± 1.3) scores vs. (6.9 ± 1.4) scores, and there were statistical differences (P<0.05). During the follow-up period, none of the patients had recurrent symptoms and obvious adverse reaction. Conclusions Percutaneous tibial nerve stimulation is an effective way to treat chronic pelvic pain syndrome.

Extracorporeal membrane oxygenation(ECMO) is a type of technique which can be used to replace patients' pulmonary function and cardiac function effectively. Since 2000s,the technique began to apply in pediatric patients but still developed very slowly. As well,patients transport with ECMO is another problem for the pediatricians. So,it is necessary to raise the conception of pediatric ECMO transport network and construction of the network. In this paper,the concept of pediatric ECMO transport network,current status of transport,the characteristics and constitution of pediatric ECMO transport network,indication for transport of pediatric ECMO,the problems in the construction of network were introduced to promote the construction of the network.