Objective: To investigate the mechanism of action of Prim-O-glucosylcimifugin (POG) to improve cholesterol metabolism in osteoarthritic (OA) chondrocytes based on the long noncoding RNA nuclear-enriched transcript 1 (lncRNA NEAT1)/microRNA-128-3p (miR-128-3p) pathway. Methods: For in vivo experiments, 60 mice were divided into the normal, sham operation, model, and POG groups using the random number table method, with 15 mice per group. The osteoarthritis mouse model was constructed using the modified Hulth method in the model and POG groups. Mice in the POG group were administered 30 mg/(kg·d)POG by gavage. The other groups were administered an equal amount of normal saline for 8 weeks. The cartilage tissue structure of mice in each group was observed using hematoxylin and eosin staining. Real-time PCR was used to detect changes in the lncRNA NEAT1 and miR-128-3p mRNA expression levels in the cartilage tissues of mice. Western blotting was used to detect the protein expressions of ATP-binding cassette transporter A1 (ABCA1), liver X receptor β (LXRβ), matrix metalloprotein-3 (MMP-3), and B-lymphoblastoma-2-associated X protein (Bax) in articular cartilage of mice. An enzyme-linked immunosorbent assay was used to measure the tumor necrosis factor-α (TNF-α) content in the synovial fluid of mice. A biochemical microplate assay was used to measure the total cholesterol level in the synovial fluid of mice. The in vitro experiments were divided into the negative control, interleukin-1β(IL-1β), IL-1β+ POG, IL-1β+ oe-lncRNA NEAT1, IL-1β+ oe-lncRNA NEAT1 + POG, IL-1β + miR-128-3p inhibition, and IL-1β+ miR-128-3p inhibition+ POG groups. An OA model was established by inducing chondrocytes with IL-1β for 24 h, and 90 mg/L of POG and miR-128-3p inhibitor(50 nmol/L) were administered for 48 h as an intervention. lncRNA NEAT1 expression in chondrocytes was detected using fluorescence in situ hybridization. A dual luciferase assay was used to detect the targeting relationship between lncRNA NEAT1 and miR-128-3p. Lentiviral plasmids overexpressing lncRNA NEAT1 were used to transfect mouse chondrocytes. Real-time PCR was used to detect the effect of lncRNA NEAT1 overexpression on the mRNA level of miR-128-3p in chondrocytes. Western blotting was used to detect ABCA1, LXRβ, MMP-3, and Bax protein expression in chondrocytes after lncRNA NEAT1 overexpression and miR-128-3p inhibition. Results: POG significantly reduced OA cartilage tissue damage. Compared with the model group, the lncRNA NEAT1 mRNA level decreased, whereas the miR-128-3p mRNA level increased in the cartilage tissue of the POG group (P<0.05). Compared with the model group, ABCA1 and LXRβ protein expression increased in the POG group, whereas MMP-3 and Bax protein expression decreased (P<0.05). The TNF-α levels decreased in the POG group compared to the model group (P<0.05). Compared with the model group, the total cholesterol level in the synovial fluid of the joint of mice in the POG group decreased (P<0.05). The mean fluorescence intensity of lncRNA NEAT1 in the IL-1β+ POG group decreased compared with the IL-1β group (P<0.05). The relative luciferase activity in the miR-128-3p mimics group bound to the lncRNA NEAT1-WT plasmid decreased compared with the miR-128-3p negative control group (P<0.05). The lncRNA NEAT1 mRNA levels decreased, whereas the miR-128-3p mRNA levels increased in the IL-1β+ oe-lncRNA NEAT1 + POG group compared with the IL-1β+ oe-lncRNA NEAT1 group (P<0.05). Compared with the IL-1β+ POG group, ABCA1 and LXRβ protein expression decreased, whereas MMP-3 and Bax protein expression increased (P<0.05). Conclusion POG mediates lncRNA NEAT1/miR-128-3p to improve cholesterol metabolism in OA chondrocytes.

ObjectiveTo develop a standardized living will manual suitable for promotion in China， combining existing literature and the current implementation status of living will both domestically and internationally as well as the characteristics of modern medicine. MethodsA pool of manual entries was constructed through hospital interviews and literature retrieval. A Delphi method expert correspondence was conducted to modify and supplement the manual entry settings. The results of the correspondence were statistically analyzed to verify the reliability and scientific validity of the manual and the correspondence. ResultsThis study constructed a pool of manual entries by conducting interviews in four hospitals in Beijing and retrieving and including 155 pieces of literature，and developed a preliminary version of the manual with a total of 40 entries in four sections， namely medicine， psychology， society， and farewell. The manual items were revised through two rounds of the Delphi method correspondence with a total of 88 experts. The expert response rates in the two rounds of the correspondence questionnaires were 100% and 90%， respectively； the expert authority coefficients were 0.84 and 0.865， respectively； and Kcndall W was 0.141 and 0.077， respectively. In terms of the reliability test， the Cronbach’s α coefficients of the two rounds of correspondence were 0.941 and 0.969， respectively， and the Guttman coefficients were 0.862 and 0.857， respectively. As for the validity test， the Pearson correlation coefficient for 39 entries was R>0.4 in the first round of correspondence， and for 41 entries was R>0.4 in the second round of correspondence； a total of 20 entries in the first round of correspondence had an I-CVI ≥ 0.780， and the scale’s S-CVI was 0.786； a total of 31 entries in the second round of correspondence had an I-CVI ≥ 0.780， and the scale’s S-CVI was 0.846. Meanwhile， this study developed a legal instrument section of the manual including personal information， effective time， modification and revocation， witnesses， and other contents by reviewing relevant laws and regulations both domestically and internationally. ConclusionThe standardized living will manual is comprehensive， reliable， and scientific， which makes it suitable for promotion and further improvement.

ObjectiveTo explore whether the mechanism of Shuangshen Ningxin capsules （SSNX） in alleviating myocardial ischemia-reperfusion injury （MIRI） in rats is related to the regulation of mitochondrial fission and fusion. MethodThis study focused on Sprague Dawley （SD） rats and ligated the left anterior descending branch of the coronary artery to construct a rat model of MIRI. The rats were divided into the sham operation group， model group， SSNX group （90 mg·kg^-1） and trimetazidine group （5.4 mg·kg^-1）. The activity of superoxide dismutase （SOD） and the content of malondialdehyde （MDA） were detected by micro method. Changes in mitochondrial membrane potential （△Ψm） and the degree of mitochondrial permeability transition pore （mPTP） opening were detected by the chemical fluorescence method. The intracellular adenosine triphosphate （ATP） level was detected by the luciferase assay. The messenger ribonucleic acid （mRNA） and protein expression levels of mitochondrial fission and fusion related factors dynamin-related protein 1 （DRP1）， mitochondrial fission 1 protein （FIS1）， optic atrophy protein 1 （OPA1）， mitochondrial outer membrane fusion protein 1 （MFN1）， and MFN2 were detected by real-time polymerase chain reaction （real-time PCR） and Western blot. ResultCompared with the sham operation group， the model group showed a decrease in serum SOD activity and an increase in MDA content. The opening level of mPTP， the level of △Ψm and ATP content decreased， the protein expressions of mitochondrial fission factors DRP1 and FIS1 increased， and the protein expressions and mRNA transcription levels of fusion related factors OPA1 and MFN1 decreased. Compared with the model group，SSNX significantly increased serum SOD activity， reduced MDA content， increased intracellular ATP level and △Ψm， reduced the opening level of mPTP， downregulated the protein expressions of mitochondrial fission factors DRP1 and FIS1， and increased the mRNA transcription levels and protein expressions of fusion related factors OPA1 and MFN1. ConclusionSSNX inhibits the expressions of mitochondrial fission factors DRP1 and FIS1， and increases the expressions of fusion related factors OPA1 and MFN1， inhibiting mitochondrial fission and increasing mitochondrial fusion， thereby alleviating MIRI.

This study aims to investigate the effect of salvianolic acid B (Sal B), the active ingredient of Salvia miltiorrhiza, on H9C2 cardiomyocytes injured by oxygen and glucose deprivation/reperfusion (OGD/R) through regulating mitochondrial fission and fusion. The process of myocardial ischemia-reperfusion injury was simulated by establishing OGD/R model. The cell proliferation and cytotoxicity detection kit (cell counting kit-8, CCK-8) was used to detect cell viability; the kit method was used to detect intracellular reactive oxygen species (ROS), total glutathione (t-GSH), nitric oxide (NO) content, protein expression levels of mitochondrial fission and fusion, apoptosis-related detection by Western blot. Mitochondrial permeability transition pore (MPTP) detection kit and Hoechst 33342 fluorescence was used to observe the opening level of MPTP, and molecular docking technology was used to determine the molecular target of Sal B. The results showed that relative to control group, OGD/R injury reduced cell viability, increased the content of ROS, decreased the content of t-GSH and NO. Furthermore, OGD/R injury increased the protein expression levels of dynamin-related protein 1 (Drp1), mitofusions 2 (Mfn2), Bcl-2 associated X protein (Bax) and cysteinyl aspartate specific proteinase 3 (caspase 3), and decreased the protein expression levels of Mfn1, increased MPTP opening level. Compared with the OGD/R group, it was observed that Sal B had a protective effect at concentrations ranging from 6.25 to 100 μmol·L^-1. Sal B decreased the content of ROS, increased the content of t-GSH and NO, and Western blot showed that Sal B decreased the protein expression levels of Drp1, Mfn2, Bax and caspase 3, increased the protein expression level of Mfn1, and decreased the opening level of MPTP. In summary, Sal B may inhibit the opening of MPTP, reduce cell apoptosis and reduce OGD/R damage in H9C2 cells by regulating the balance of oxidation and anti-oxidation, mitochondrial fission and fusion, thereby providing a scientific basis for the use of Sal B in the treatment of myocardial ischemia reperfusion injury.

Hepatocellular carcinoma (HCC) is one of the most common malignant tumors in the digestive tract system, which is induced by multiple factors, involving multiple genes and complicated mechanism. Its incidence and mortality rank fourth and second respectively in China, and accounting for more than 85% of primary liver cancers. Tumor immune microenvironment (TIME), plays a critical role in determining the tumor progression and treatment outcomes, making it become a hotspot in current studies. Summarising the previous studies, it is found that the progression of HCC is significantly influenced by the TIME and its complex interactions. TIME consists of various cellular and non-cellular components, such as myeloid-derived suppressor cells (MDSCs), tumor-associated macrophages (TAMs), tumor-associated neutrophils (TANs), regulatory T cells (Tregs), innate lymphoid cells (ILCs), as well as growth factors, proteolytic enzymes, and extracellular matrix proteins. Due to long-term exposure to bacterial components carried by the portal vein, food-derived antigens, and a large amount of foreign antigenic substances, the microenvironment of liver exhibits a certain degree of immune suppression to resist excessive inflammation caused by the non-pathogenic intestinal environment. Besides, the inhibitory immune microenvironment shaped by tumor cells which induces changes in the phenotype and function of immune cells, and attenuates the cytotoxic capabilities of immune system. Meanwhile, the regulation of immune cell metabolism is crucial for anti-tumor immune response. Abnormal metabolites of liver cancer microenvironment and intestinal flora metabolites regulate the remodeling of immune microenvironment and the progression in liver cancer. Normally, the cancer immune cycle functions effectively to remove tumor cells, while the immunosuppressive, exhausted T cells and metabolic disorders of the TIME leads to defects in the cancer immunity cycle and promotes to tumor progression. Furthermore, during the processes of rapid proliferation and differentiation, tumor cells alter their metabolic status through “metabolic reprogramming”, allowing them to compete with anti-tumor immune cells for vital nutrients including glucose, lipids, and nucleotides. At the same time, the abnormal consumption of metabolites leads to local hypoxia, lower pH levels, and the accumulation of metabolic products, which in turn suppress the proliferation and effector functions of immune cells, ultimately facilitating immune evasion and tumor progression. According to the above, local immune imbalance and metabolic disorders in the liver collectively shape the unique microenvironment of HCC, resulting in the accumulation of immunosuppressive cytokines, extracellular matrix and abnormal metabolites. These factors induce abnormal tumor angiogenesis, recruitment of immunosuppressive cells, reduce T-cell infiltration, and diminish anti-tumor function, which accelerates the progression of HCC and immune escape. Currently, there are still remarkable limitations in the clinical treatment methods and outcomes for HCC, while immunotherapy offers a new strategy. The advantages of immunotherapy demonstrate relatively higher specificity and fewer side effects compared to traditional treatment methods such as surgery, radiotherapy, and chemotherapy. Up to now, more and more evidence has been uncovered that liver cancer immunotherapy is closely related to TIME. Targeting the TIME of HCC provides a new perspective into a deeper understanding of the mechanisms of immunotherapy resistance and the development of new immunotherapy approaches. However, single immunotherapy has not shown satisfactory results in improving the prognosis of HCC patients. At present, dual immune checkpoint inhibitors or their combination with existing therapies are being widely explored in clinical studies, hoping to overcome the limitations of HCC therapy. Therefore, this review summarizes the composition of immunosuppressive microenvironment in liver cancer and metabolic regulation, and further discusses clinical therapeutic strategies by targeting microenvironment remodeling for the treatment of liver cancer, which provides new avenues for tumor immunotherapy.

Objective To investigate the mechanism by which Tougu Xiaotong Capsule(TGXTC)alleviates chondrocyte degeneration in knee osteoarthritis(KOA).Methods Thirty 2-month-old C57BL/6 mouse models of KOA established using the Hulth method were randomized into model group,TGXTC group,and diclofenac sodium group and received treatment with saline,TGXTC(368 mg/kg),and diclofenac sodium(10 mg/kg)by gavage,respectively,with another 10 untreated mice as the blank control group.All interventions were administered 6 times a week for 4 weeks.After the treatments,structural changes in the cartilage tissue were observed with morphological staining,and Nav1.7 mRNA expression and the protein expression levels of Nav1.7,MMP-3,ADAMTS-5,and COX-2 were detected using RT-qPCR and Western blotting.Fluorescence in situ hybridization(FISH)was used to detect Nav1.7 expression in the chondrocytes.In cultured KOA chondrocytes,the effect of TGXTC and lentivirus-mediated Nav1.7 knockdown on MMP-3,MMP-13,ADAMTS-4,ADAMTS-5,and COX-2 protein expressions were assessed with Western blotting.Results In KOA mice treatments with TGXTC and diclofenac sodium both significantly alleviated structural damage of the cartilage layer,reduced Nav1.7 protein and mRNA expressions and lowered the expressions of MMP-3,ADAMTS-5,and COX-2 proteins in the cartilage tissues.FISH results indicated that TGXTC treatment significantly reduced IL-1β-induced Nav1.7 expression in the chondrocytes.In Nav1.7 knockdown experiment,Nav1.7 levels were significantly lower in IL-1β+sh-Nav1.7 group than in IL-1β group,and also lower in IL-1β+TGXTC group than in IL-1β+sh-Nav1.7+TGXTC group.TGXTC treatment significantly inhibited IL-1β-induced elevation of MMP-3,MMP-13,ADAMTS-4,ADAMTS-5 and COX-2 protein expressions in the chondrocytes,but its effects were strongly weakened by Nav1.7 knockdown.Conclusion TGXTC alleviates extracellular matrix metabolic disorder in KOA chondrocytes by regulating Nav1.7,thereby mitigating chondrocyte degeneration in KOA mice.

Objective:To evaluate the detective effect of double inversion recovery (DIR) sequence on cerebral white matter hyperintensities (WMH) in patients with acute ischemic stroke, and compare with those of T2WI and FLAIR sequences.Methods:Seventy-three acute ischemic stroke patients with WMH within 14 d of onset, admitted to Department of Neurology, Beijing Shijitan Hospital, Capital Medical University from November 2018 to March 2021, were chosen. MRI T2WI, FLAIR and DIR sequences were used to detect WMH. According to Fazekas scale, patients with periventricular white matter hyperintensities (PVWMH) or deep white matter hyperintensities (DWMH) were divided into mild group (score of 0-1) and moderate to severe group (scores≥2); the differences in WMH volume detected by T2WI, FLAIR and DIR sequences, and signal intensity, cross-sectional area and contrast of isolated lesions were compared.Results:(1) Seventy-three patients were with PVWMH (36 into the mild group and 37 into the moderate to severe group); in patients from the moderate to severe group, PVWMH volume detected by FLAIR sequence was statistically larger compared with that by DIR sequence, and PVWMH volume detected by T2WI sequence was significantly smaller compared with that by FLAIR sequence ( P<0.05). Fifty-seven patients were with DWMH (44 into the mild group and 13 into the moderate to severe group); the DWMH volume detected by FLAIR and T2WI sequences was significantly larger than that by DIR sequence ( P<0.05). (2) A total of 60 isolated lesions were detected, ranged 5.0-9.1 mm in length; isolated lesions enjoying significantly larger cross-sectional area, higher signal intensity, and lower contrast detected by FLAIR and T2WI sequences compared with those by DIR sequence ( P<0.05); isolated lesions enjoying significantly higher signal intensity and contrast detected by T2WI sequence compared with those by FLAIR sequence ( P<0.05). Conclusion:DIR sequence enjoys better effect in detecting WMH than FLAIR and T2WI sequences; the mismatch area of DIR sequence with FLAIR or T2WI sequences suggests WMH penumbra.

Objective:This study aimed to provide basic data for the prevention, diagnosis and treatment of pediatric viral myocarditis (VMC) in China through analyzing the epidemic characteristics and disease burden of pediatric inpatients with VMC from 2016 to 2021.Methods:We performed a descriptive statistical analysis to the age, genders, seasons, regions and hospitalization cost and days of pediatric VMC inpatients and the death. All of the information was obtained from 27 Children′s hospitals or Maternal and Child Health hospitals of 23 provinces of China from 2016 to 2021.Results:A total of 7 647 599 cases including 1 646 VMC inpatients were admitted into our study. The annual numbers of hospitalizations were 173, 227, 313, 301, 295 and 337, with the hospitalized constituent ratios being 14.9/100 000, 17.9/100 000, 23.0/100 000, 20.5/100 000, 26.5/100 000 and 26.4/100 000 from 2016 to 2021. In recent 6 years, the proportion of VMC hospitalizations had increased yearly ( P<0.001), and had associated with the onset age ( P<0.001). Aged 12-≤18 years owned the highest hospitalized constituent ratio. The Northeast of China owned the largest number of VMC inpatients, and the East second to it. Among the 1 646 VMC children, there were 68 deaths, with the hospitalized case fatality rate of 4.13%. There were no significant differences between genders, age, seasons, years and fatality rate of VMC inpatients. For the diseases burden, the median of hospitalization days of all VMC inpatients was 10 days (IQR 6, 21), and the median of hospitalization cost was 1 1 842.3 RMB (IQR 6 969.22, 19 714.78). The median of hospitalization days of deceased VMC children was only 1 day (IQR 1, 3), the median cost could be 8 874.03 RMB (IQR 5 277.94, 5 6 151.59). Conclusions:In this study, we found that proportion of hospitalization of VMC children increased year by year, adolescence might be a risk factor of VMC. The fatality of VMC inpatients could be up to 4.13%, and the death led to a huge economic burden of society, family and individuals.

β-receptor blockers are used to treat cardiovascular disease(CVD),while β-receptor agonists are used to treat chronic obstructive pulmonary disease(COPD).The two drugs have opposite mechanisms of action.Howev-er,there is a close relationship between COPD and CVD,and CVD is related to the severity of pulmonary dysfunc-tion.The use of β-receptor blockers in COPD is still controversial,but studies have shown that cardiac selective β-receptor blockers can reduce mortality in mild and moderate COPD with CVD and reduce the risk of COPD exacerbation,while non-selective β-receptor blockers increase the risk of COPD exacerbation.However,the dosage of cardiac selective β-receptor blockers and its balance with β-receptor agonists in COPD patients with CVD still requires serious treatment.Research on new β-receptor blockers is a problem to be paid close attention to in the future.

Objective:To explore the risk factors for lymphoproliferative disorders (PTLD) in children with thalassemia major (TM) after allogeneic hematopoietic stem cell transplantation (allo-HSCT).Methods:This was a retrospective case-control study.A total of 482 children with TM who underwent allo-HSCT at Shenzhen Children′s Hospital between January 2020 and December 2022 were selected and classified into the PTLD and non-PTLD groups according to the occurrence of PTLD.The risk factors for PTLD after allo-HSCT in children with TM were analyzed, and the diagnostic efficiency of relevant risk factors for PTLD was analyzed by receiver operating characteristic (ROC) curve.Results:A total of 25 out of 482 patients (5.2%, 25/482) developed PTLD about 114 (54-271) days after allo-HSCT.Among them, 12 cases (12/25, 48.0%) occurred within 100 days, and 22 cases (22/25, 88.0%) occurred within 1 year after allo-HSCT.Univariate analysis showed that there were significant differences in gender composition, type of transplant donor, number of natural killer cells and B lymphocytes in peripheral blood at 30 days after allo-HSCT, positive rate of plasma Epstein-Barr virus deoxyribonucleic acid (EBV-DNA) and incidence rate of acute graft-versus-host disease (aGVHD) between the 2 groups (all P<0.05).Multivariate Logistic regression analysis showed that female ( OR=3.196, 95% CI: 1.144-8.929), positive plasma EBV-DNA ( OR=17.523, 95% CI: 5.449-56.344) and aGVHD ( OR=3.156, 95% CI: 1.161-8.575) were independent risk factors for PTLD after allo-HSCT in TM children (all P<0.05).The ROC curve analysis showed that positive plasma EBV-DNA had an excellent accuracy in predicting the occurrence of PTLD after allo-HSCT (sensitivity was 0.796, specificity was 0.800, area under the curve was 0.803).If combined with aGVHD and gender, the area under the curve for the prediction of PTLD increased to 0.831. Conclusions:Female, positive plasma EBV-DNA and aGVHD are independent risk factors for PTLD after allo-HSCT in children with TM.It provides useful early warnings for the prediction and prevention of PTLD.