OBJECTIVE: Myocardial inflammation during myocardial infarction (MI) could be inhibited by regulating arachidonic acid (AA) metabolism. Recent studies demonstrated that Sini Decoction (SND) was identified to be an effective prescription for treating heart failure (HF) caused by MI. But the anti-inflammatory mechanism of SND remained unclear. The work was designed to investigate the anti-inflammatory mechanism of SND through the AA metabolism pathway in vitro and in vivo experiments. METHODS: An inflammatory injury model of H9c2 cells was established by lipopolysaccharide (LPS)-stimulated macrophage-conditioned medium (CM). The MI model was built by the ligation of left anterior descending (LAD) branch of coronary artery in rat. Meanwhile, the rats were divided into five groups: sham group, MI group, MI + Celecoxib group, MI + low-dose SND group (SND-L) and MI + high-dose SND group (SND-H). Cardiac function, histopathological changes and serum cytokines were examined four weeks later. Western blot analysis was conducted to verify the key enzymes levels in the AA metabolic pathway, including phospholipase A2 (PLA2), cyclooxygenases (COXs) and lipoxygenases (LOXs). RESULTS: These in vivo results demonstrated that SND could improve the cardiac function and pathological changes of rats with MI, and regulate the key inflammatory molecules in the AA metabolism pathway, including sPLA2, COX-1, COX-2, 5-LOX and 15-LOX. In vitro, SND could decrease the release of pro-inflammatory cytokines including TNF-α and IL-6 and inhibit cell apoptosis in CM-induced H9c2 cells. Moreover, SND could protect H9c2 cells from the damage of CM by regulating nuclear factor kappa-B (NF-κB) signal pathway and the expression of COX-2. CONCLUSION SND may be a drug candidate for anti-inflammatory treatment during MI by regulating the multiple targets in the AA metabolism pathway.

Extensive epigenetic reprogramming involves in memory CD8+ T-cell differentiation. The elaborate epigenetic rewiring underlying the heterogeneous functional states of CD8+ T cells remains hidden. Here, we profile single-cell chromatin accessibility and map enhancer-promoter interactomes to characterize the differentiation trajectory of memory CD8+ T cells. We reveal that under distinct epigenetic regulations, the early activated CD8+ T cells divergently originated for short-lived effector and memory precursor effector cells. We also uncover a defined epigenetic rewiring leading to the conversion from effector memory to central memory cells during memory formation. Additionally, we illustrate chromatin regulatory mechanisms underlying long-lasting versus transient transcription regulation during memory differentiation. Finally, we confirm the essential roles of Sox4 and Nrf2 in developing memory precursor effector and effector memory cells, respectively, and validate cell state-specific enhancers in regulating Il7r using CRISPR-Cas9. Our data pave the way for understanding the mechanism underlying epigenetic memory formation in CD8+ T-cell differentiation.
CD8-Positive T-Lymphocytes/metabolism* ; Cell Differentiation ; Chromatin/immunology* ; Animals ; Mice ; Immunologic Memory ; Epigenesis, Genetic ; SOXC Transcription Factors/immunology* ; NF-E2-Related Factor 2/immunology* ; Mice, Inbred C57BL ; Gene Regulatory Networks ; Enhancer Elements, Genetic

Objective To review the diagnostic information of 3368 viral hepatitis B inpatient cases reported by some healthcare institutions in Qinghai Province in the National Notifiable Disease Report System(NNDRS)in 2021,and to analyse the consistency of their reports and influence factor.Methods The study used the typical survey method to study 3368 hospitalized cases of hepatitis B reported by 32 medical institutions in Qinghai Province in 2021 from July 2022 to April 2023.The consistency between the reported classification and the reviewed classification was evaluated by reviewing diagnostic information of the cases,combined with the Kappa consistency test,and the consistency-influencing factors were analysed in a univariate and multivariate analysis.Results The consistency between acute,chronic,and unclassified reporting classifications and review classifications was poor(Kappa＜0.40);the consistency between acute and chronic review was poor(Kappa＜0.40).The main influence factor of the consistency of review included the level of medical institution,the patient's gender,occupation,past history,and the unit of diagnosis of initial onset of illness.Provincial medical institutions reported higher consistency,and male cases and herder cases had better consistency than female cases and other occupations.Review consistency of cases with past history was better than that of cases with unclear past history.Conclusion The consistency of diagnosis and reporting of hepatitis B cases in healthcare institutions in Qinghai Province in 2021 was low,with problems of inaccurate classification and duplicate reporting.It is recommended to strengthen the training and supervision of healthcare institutions,standardise the filling of'attached card'information,and promote the hepatitis B core antibody IgM test to improve the classification and diagnosis of acute and chronic hepatitis B,so as to comprehensively improve the accuracy of the diagnosis and reporting of hepatitis B cases.

Objective:To explore the relationship between Epstein-Barr virus (EBV) infection, hepatitis B virus (HBV) reactivation and clinical features and prognosis in HBV-infected non-Hodgkin lymphoma (NHL) patients and influencing factors of HBV reactivation.Methods:A retrospective cohort study was conducted. A total of 80 NHL patients with hepatitis B surface antigen (HBsAg) positive (which was defined as HBV positive) who were admitted to the Second People's Hospital of Lianyungang and Jiangsu Province Hospital from December 2012 to October 2022 were selected. All patients were divided into EBV-positive group and EBV-negative group according to EBV DNA results, and further grouped into the HBV reactivation group and the non-reactivation group according to whether HBV were reactivated after chemotherapy. The clinical characteristics of patients among groups were compared. Multivariate logistic regression model was used to analyze the factors influencing HBV reactivation. The Kaplan-Meier method was used to evaluate the progression-free survival (PFS) and overall survival (OS) of patients, and the log-rank test was used for inter-group comparison.Results:Among NHL patients with HBV positive, 27 cases (33.8%) were EBV-positive and 29 cases (36.3%) were HBV reactivation. Compared with the EBV-negative group, the proportion of patients with Ann Arbor stage Ⅲ-Ⅳ [92.6% (25/27) vs. 66.0% (35/53)], elevated β 2-microglobulin level [88.9% (24/27) vs. 62.3% (33/53)], bone marrow involvement [40.7% (11/27) vs. 15.1% (8/53)], and HBV reactivation [51.9% (14/27) vs. 28.3% (15/53)] was higher in the EBV-positive group, and the differences were statistically significant (all P<0.05). There were no statistically significant differences in the composition of patients stratified by age, gender, pathological type, B symptom, lactate dehydrogenase level, international prognostic index score, number of extranodal involvements, liver involvement, hepatitis outbreak, prophylactic anti-HBV therapy, hepatitis B surface antibody (HBsAb), rituximab therapy, and the last chemotherapy effects between the 2 groups (all P > 0.05). Compared with the HBV non-reactivation group, the proportion of patients undergoing hepatitis outbreak [48.3% (14/29) vs. 17.6% (9/51)], not receiving prophylactic anti-HBV therapy [65.5% (19/29) vs. 39.2% (20/51)], HBsAb negative [79.3% (23/29) vs. 21.6% (11/51)], EBV positive [48.3% (14/29) vs. 25.5% (13/51)], receiving rituximab [82.8% (24/29) vs. 60.8% (31/51)] was higher in the HBV reactivation group, and the differenves were statistically significant (all P < 0.05); while there were no statistically significant differences in the composition of patients stratified by the other clinical characteristics between the 2 groups (all P > 0.05). Multivariate logistic regression analysis showed that EBV-positivity was an independent risk factor for HBV reactivation after chemotherapy in NHL patients with HBsAg positive ( OR = 7.073, 95% CI: 1.613-31.010, P = 0.009), while HBsAb positive ( OR = 0.038, 95% CI: 0.008-0.186, P < 0.001) and preventive anti-HBV therapy ( OR = 0.172, 95% CI: 0.039-0.756, P = 0.020) were independent protective factors. The last follow-up was in December 2023 and the median follow-up time was 36.5 months. There were no statistically significant differences in PFS and OS between the EBV-positive group and the EBV-negative group, HBV reactivation group and the non-reactivation group (all P > 0.05). Conclusions:Among HBV-infected NHL patients, those with concurrent EBV infection have a more advanced clinical stage and are very prone to bone marrow invasion, and they also show a higher probability of HBV reactivation; HBV reactivation may be related to whether receiving preventive anti-HBV therapy and rituximab therapy. EBV infection may increase the risk of HBV reactivation in NHL patients; EBV infection and HBV reactivation may not be relevant to the prognosis of patients.

Acute T-lymphoblastic leukemia (T-ALL) is a hematopoietic malignancy, and in recent years, with the advancement of combined chemotherapy and hematopoietic stem cell transplantation, the prognosis of T-ALL has improved significantly, but for patients with primary drug resistance or relapsed/refractory disease the prognosis is still poor. The plant homeodomain finger 6 (PHF6) is a tumor suppressor protein, it plays a pivotal role in T cell differentiation, epigenetic regulation and oncogenic pathway synergy, and its mutations and deletions are commonly associated with the development of T-lymphocytic leukemia. However, the underlying mechanism of PHF6 in the pathogenesis of T-ALL remains unclear. This article reviews the structure, function and mechanism of action of PHF6 in T-ALL, the important coexisting genes associated with the progression of T-ALL, and the research progress in targeted therapy.

Asthma is essentially a chronic inflammatory disease of the airways,and the proposed"gut-lung axis"provides a new idea for exploring its pathogenesis and therapeutic targets.A number of studies have confirmed that gut microbiota dysbiosis may affect the immune-inflammatory response through metabolites,which were involved in the disease process of asthma.Fecal microbiota transplantation(FMT)is a method that can efficiently reconstruct the gut microbiota of patients.It has been used to treat gastrointestinal diseases,central nervous system diseases,inflammatory diseases,and so on.Standardized operational protocols have been established.The use of probiotics or prebiotics in treating and preventing asthma and applying FMT in mice models of asthma have pointed the way for new prevention and treatment strategies.This article reviews the relationship between gut microbiota and asthma,and the feasibility of FMT in treating and preventing asthma.

Objective To observe the clinical efficacy of fire needling in treating lumbar disc herniation(LDH)of cold-damp type under the guidance of meridian sinew theory.Methods A total of 70 cases of patients with LDH of cold-damp type were randomly divided into observation group and control group,35 cases in each group,the control group was given conventional acupuncture treatment,and the observation group was treated with fire needling under the guidance of meridian sinew theory on the basis of the treatment of the control group,and the course of treatment covered three courses,one week as a course.After three courses of treatment,the clinical efficacy of the two groups was evaluated,and the changes in the Visual Analogue Scale(VAS)scores of pain,the Japanese Orthopedic Association(JOA)scores,and traditional Chinese medicine(TCM)syndrome scores of patients in the two groups before and after treatment were observed.Results(1)During the study,there were three cases in the observation group and five cases in the control group were failed to follow-up.Finally,32 cases in the observation group and 30 cases in the control group were included in the efficacy statistics.(2)The total effective rate was 93.75%(30/32)in the observation group and 86.67%(26/30)in the control group.The clinical efficacy of the observation group was superior to that of the control group,and the difference being statistically significant(P<0.05).(3)After treatment,the VAS and JOA scores of patients in the two groups improved significantly(P<0.05),and the improvement in the observation group was significantly superior to that in the control group,the difference being statistically significant(P<0.05).(4)After treatment,the TCM scores of the two groups of patients improved significantly(P<0.05),and the improvement in the observation group was significantly superior to that in the control group,the difference being statistically significant(P<0.05).Conclusion Under the guidance of the meridian sinew theory,fire needling in treating LDH of cold-damp type can significantly reduce the degree of pain,improve the lumbar function,alleviate LDH-related symptoms,so as to improve the patients'quality of life.

ObjectiveTo investigate the possible mechanism by which Zhiqiao Gancao Decoction （枳壳甘草汤， ZGD） delays intervertebral disc degeneration （IDD） based on the Hippo-yes-associated protein （YAP）/transcriptional co-activator with PDZ-binding motif （TAZ） signaling pathway. MethodsA total of 50 SD rats were randomly divided into sham surgery group， model group， low-dose ZGD group， high-dose ZGD group， and high-dose ZGD + inhibitor group， with 10 rats in each group. In the sham surgery group， the rats were pierced in the skin and muscle at the Co6/7/8 segments of the tail with a 21G needle （depth approximately 2 mm） without damaging the intervertebral disc. In the other groups， rats were injected with a 21G needle at the Co6/7/8 segments of the tail to establish an IDD model by piercing the tail intervertebral disc 5 mm. One week after modeling， rats in the low-dose and high-dose ZGD groups were given 6.24 and 12.24 g/（kg·d） of the decoction via gastric gavage， respectively. The high-dose ZGD + inhibitor group was given 12.24 g/（kg·d） of the decoction and an intraperitoneal injection of YAP/TAZ inhibitor Verteporfin 10 mg/kg. The sham surgery and model groups were given 5 ml/（kg·d） of normal saline via gavage. The gavage was given once a day， and the intraperitoneal injection was given every other day. After 4 weeks of continuous intervention， the pathological changes of the tail intervertebral discs were observed using HE staining， Oil Red O-Green staining， and Toluidine Blue staining. Immunohistochemistry was used to detect the expression of aggrecan and MMP3 in the nucleus pulposus. TUNEL fluorescence staining was performed to detect apoptosis in the nucleus pulposus， and the apoptosis rate was calculated. Western blot was used to detect the Hippo-YAP/TAZ signaling pathway， including YAP， phosphorylated YAP （p-YAP）， phosphorylated MST1/2 （p-MST1/2）， phosphorylated TAZ （p-TAZ） and apoptosis-related proteins， such as Cleaved Caspase 3， P53， Bcl-2 and Bax. ResultsCompared with sham surgery group， the rats in the model group showed significant degenerative changes in the intervertebral disc. The levels of aggrecan， Bcl-2， and YAP proteins in the nucleus pulposus decreased， while the levels of p-MST1/2， p-YAP， p-TAZ， P53， Bax， Cleaved Caspase 3， MMP3 proteins， and the apoptosis rate increased （P < 0.01）. Compared with the model group， the drug intervention groups showed partial recovery in intervertebral disc degeneration. The levels of aggrecan， Bcl-2， and YAP proteins increased， while the levels of p-MST1/2， p-YAP， p-TAZ， P53， Bax， Cleaved Caspase 3， MMP3 proteins， and the apoptosis rate decreased （P＜0.05 or P＜0.01）. The high-dose ZGD group showed more significant recovery in intervertebral disc degeneration compared to the low-dose ZGD group， with a decrease in the levels of p-MST1/2， p-YAP， p-TAZ， P53， Bax， Cleaved Caspase 3， MMP3 proteins， and apoptosis rate， and an increase in the levels of aggrecan， Bcl-2， and YAP proteins （P＜0.05 or P＜0.01）. Compared with the high-dose ZGD group， the high-dose ZGD + inhibitor group showed a reduced recovery in intervertebral disc degeneration， with an increase in the levels of p-MST1/2， p-YAP， p-TAZ， P53， Bax， Cleaved Caspase 3， MMP3 proteins， and apoptosis rate， and a decrease in the levels of aggrecan， Bcl-2， and YAP proteins （P＜0.05 or P＜0.01）. ConclusionZGD may delay intervertebral disc degeneration by inhibiting the phosphorylation of YAP in the nucleus pulposus， maintaining the function of the Hippo-YAP/TAZ signaling pathway， and reducing apoptosis of nucleus pulposus cells.

Objective To discuss the preliminary application of CRISPR-Cas9 gene editing technology in repair of antithrombin gene(SERPINC1)c.318_319insT mutation.Methods The single guide RNA(sgRNA)was designed by CRISPR online design website,and AT c.318_319 insT mutant and CRISPR-Cas9 repairsome were constructed.The gene fragments from the wild-type gene,AT c.318_319 insT mutant and CRISPR-Cas9 repairsome were transferred into lentiviral expression vectors,and then PCR sequencing was performed for verification.The successfully constructed lentiviral recombinant plasmids were transfected into the human embryonic kid-ney cells(HEK293T).After cell culture,HEK293T cells were lysed.The AT:Ag levels in the cell lysing reagents from wild-type gene,CRISPR-Cas9 repairsome and mutant were compared by ELISA and Western blot,respectively.The recombinant AT protein was characterized in vitro by cellular immunofluorescence assay.Results Both the AT c.318_319insT mutant and CRISPR-Cas9 repair-some were successfully constructed.The results of experiments with HEK293T cells in vitro showed that the wild-type AT:Ag in the cell lysing reagents was set as 100％,the AT:Ag of CRISPR-Cas9 repairsome was 47％,and the AT:Ag of AT c.318_319insT was 22％,which were consistent with the results of western blot and cellular immunofluorescence assay.Conclusion The cellular experiments in vitro verified that CRISPR-Cas9 gene editing technology could effectively repair the SERPINC1 c.318_319 insT mutation in situ,which might provide the experimental support for the application of CRISPR-Cas9 gene editing technology in the gene therapy of hereditary thrombotic diseases.

Objective To compare two methods for calculating antimicrobial use density(AUD),select the more suitable AUD for monitoring and management,and achieve rational assessment of AUD.Methods A calculation model for real-time monitoring on AUD and AUD of discharged patients was established.The AUD of 13 clinical departments and the whole hospital of a hospital from January to May 2023 calculated by two calculation methods was statistically analyzed,the dispersion was compared to evaluate their applicability,the impact of two calculation methods on departmental assessment was measured.From June 2023,the real-time monitoring AUD system began to be adopted to achieve triple monitoring on departments,doctors,and drugs.From July,assessment was initia-ted,changing trend of AUD calculated by two calculation methods after control in 2023 was compared.Results AUD of real-time monitoring had a smaller department monthly dispersion compared with the AUD of discharged patients,with no significant difference among the whole hospital,making the assessment more rational.After im-plementing management based on real-time monitoring of AUD,defined daily doses(DDDs)decreased by 4.54 in June 2023,and decreased by 2.75 DDDs in the whole year of 2023.Conclusion Real-time monitoring on AUD can better reflect the actual medication level,is more applicable,and is conducive to scientific management of AUD.