ObjectiveTo investigate the epigenetic mechanism of Diwu Yanggan Capsule in improving liver regeneration microenvironment in a rat model of liver cancer by regulating DNA methylation， and to provide a basis for scientific clinical medication. MethodsA total of 48 specific pathogen-free Sprague-Dawley rats were divided into normal group， model group， and Diwu Yanggan Capsule group using a random number table， with 16 rats in each group. The Solt-Farber two-step method was used to establish a rat model of liver cancer. The rats in the Diwu Yanggan Capsule group were given Diwu Yanggan Capsule at a dose of 750 mg/kg/d by gavage， and those in the normal group and the model group were given an equal volume of normal saline by gavage. Liver tissue samples were collected from each group of rats after 16 weeks of continuous intervention； DNA methylation chips were used to analyze the change in DNA methylation in liver tissue， and gene ontology （GO） and Kyoto Encyclopedia of Genes and Genomes （KEGG） analyses were used for data analysis. In addition， the MeDIP-PCR technique was used to detect the changes in candidate differentially methylated genes such as YWHAB， ADCK2， ERLIN2， SEMA3B， and TPH2 in the liver tissue of rats， and Western blot and RT-qPCR were used to verify the expression of key methylated genes. The independent-samples t test was used for comparison of continuous data between two groups， and a one-way analysis of variance was used for comparison between multiple groups， while the least significant difference t-test was used for further comparison between two groups. ResultsThe DNA methylation chip analysis showed that compared with the normal group， the model group had significant methylation changes in the promoter region of 2 422 genes in liver tissue of rats. The GO functional enrichment analysis and the KEGG pathway enrichment analysis showed that these differentially methylated genes were significantly enriched in metabolic pathways such as steroid hormone biosynthesis and drug metabolism-cytochrome P450. Compared with the model group， the Diwu Yanggan Capsule group had significant reversal of promoter methylation in 1 650 genes， and the KEGG enrichment analysis showed that these genes were mainly involved in the pathways closely associated with cell proliferation， apoptosis， and microenvironment regulation， such as the calcium ion signaling pathway， the cAMP signaling pathway， and the extracellular factor signaling pathway. Compared with the model group， the Diwu Yanggan Capsule group had a significant increase in the promoter methylation level of the ADCK2 gene （P<0.05） and significant reductions in the promoter methylation levels of the ERLIN2 and TPH2 genes （all P<0.05）. Compared with the model group， the Diwu Yanggan Capsule group had significant reductions in the mRNA expression levels and the protein expression levels of the ADCK2 （all P<0.05）. ConclusionAbnormal DNA methylation in liver tissue participates in the development and progression of liver cancer. The effect of Diwu Yanggan Capsule on DNA methylation level is an important epigenetic mechanism for its effect in the prevention and treatment of liver cancer.

Objective: To investigate the status of tuberculin skin test (TST) results and the influencing factors among student close contacts with pulmonary tuberculosis, so as to provide the evidence for developing prevention and control strategies for pulmonary tuberculosis among students. Methods: The students aged 15 years and above who had close contact with pulmonary tuberculosis cases in Yuecheng District, Shaoxing City, Zhejiang Province, from October 2016 to December 2023 were recruited and investigated using questionnaires and TST to collect demographic information, contact history, and TST results. The influencing factors for TST positivity among student close contacts with pulmonary tuberculosis were analyzed using a multivariable logistic regression model. Results: A total of 5 507 student close contacts were investigated, including 2 982 males and 2 525 females, with a male-to-female ratio of 1.18∶1. The mean age was (19.10±1.71) years. Among them, 397 (7.21%) were technical secondary school students, 766 (13.91%) were senior high school students, 2 556 (46.41%) were junior college students, and 1 788 (32.47%) were college students or above. A total of 958 students tested positive for TST, with a positivity rate of 17.40%. The rates of general positivity, moderate positivity, and strong positivity were 10.53%, 4.98% and 1.89%, respectively. Multivariable logistic regression analysis showed that senior high school students (OR=1.473, 95%CI: 1.009-2.152) and junior college students (OR=1.467, 95%CI: 1.074-2.005), as well as those with an exposure-to-screening interval of ≥46 days (46-<61 days, OR=2.043, 95%CI: 1.478-2.826; ≥61 days, OR=1.291, 95%CI: 1.018-1.637) had a higher risk of TST positivity. Female student close contacts had a lower risk of TST positivity (OR=0.753, 95%CI: 0.649-0.873). Conclusion The TST positivity rate was relatively high, and gender, school type, and exposure-to-screening interval were influencing factors for TST positivity among student close contacts with pulmonary tuberculosis.

ObjectiveTo analyze the influencing factors for recurrence in successfully treated pulmonary tuberculosis patients with isoniazid-resistant and rifampicin-sensitive in Shaoxing City, Zhejiang Province. MethodsData on general demographic information, treatment information and drug susceptibility test results for pulmonary tuberculosis patients admitted to the designated tuberculosis medical institutions and registered in the tuberculosis information management system was collected in Shaoxing City from January 2011 to August 2024. A total of 428 patients with isoniazid resistance (including isoniazid single resistance and multiple resistance) but who were successfully treated were included in the study. Information for the recurrence after successful treatment of the patients was analyzed. The Cox proportional hazards models were used to analyze the influencing factors of recurrence in patients. ResultsAmong the 428 successfully treated patients included in the study, 31 cases (accounting for 7.24%) had recurrence by the end of the observation period, with a recurrence rate density of 1.31 per 100 person-years and a median recurrence time of 0.99 (0.08, 8.27) years. Among the relapsed population, 51.61% of the patients relapsed within one year after successful treatment. 77.42% of the patients relapsed within two years after successful treatment. Multivariate Cox regression analysis showed that when isoniazid resistance was discovered, the diagnosis classification of relapse (HR=4.115, 95%CI: 1.734‒9.767) and positive 0-month sequence smear (HR=4.457, 95%CI: 1.053‒18.866) were risk factors for recurrence after successful treatment in patients. ConclusionRegular follow-up should be strengthened for at least two years after the successful treatment of isoniazid-resistant pulmonary tuberculosis patients. Special attention should be paid to the treatment effect and regular re-examination and monitoring after the end of the treatment course of isoniazid-resistant pulmonary tuberculosis patients who have been re-treated and were sputum smear positive at baseline, so as to prevent recurrence and disease progression in high-risk populations.

【Objective】 To investigate the relationship between the initial serum ammonia level and the risk of ICU and hospital mortalities in critically ill patients without hepatic disease. 【Methods】 A retrospective cohort study was conducted among patients admitted to the eICU Collaborative Research Database (eICU-CRD) for a single admission who had serum ammonia test records within 48 hours of the first ICU admission and had no hepatic disease. The age, sex, ethnicity, Acute Physiologic and Chronic Health Evaluation Ⅳ score (APACHE Ⅳ score), treatment methods, complications, and outcomes were extracted. Univariable and multivariable Logistic regression were used to analyze the relationship between serum ammonia level and the risk of mortality. Interactions were used to analyze whether the relationship between serum ammonia level and the risk of mortality differed in subgroups of APACHE Ⅳ scores, age, sex, and ethnicity; subgroup analyses were made. 【Results】 A total of 1 674 patients were included. The multivariable Logistic regression showed that for every 10 μg/dL increase in ammonia, the risk of ICU death increased by 6.9% (OR=1.069, 95% CI: 1.036-1.104), and the risk of hospital death increased by 4.6% (OR=1.046, 95% CI: 1.017-1.076). The risk of ICU death was 1.7 times greater in patients with initial ammonia level of 49-82 μg/dL than in those with <49 μg/dL (OR=1.700, 95% CI: 1.165-2.482), the risk of ICU death was 2.862 times greater in patients with a level of ≥82 μg/dL compared to those with <49 μg/dL (OR=2.862, 95% CI: 1.792-4.570), and the risk of hospital death was 1.844 times higher in the ≥82 μg/dL group than in the <49 μg/dL group (OR=1.844, 95% CI: 1.213-2.804). There were no significant differences between initial ammonia level and the risk of mortalities in different subgroups of APACHEⅣ scores, age, sex, or ethnicity. 【Conclusion】 In critically ill patients without hepatic disease, elevated initial serum ammonia level after ICU admission is associated with a high risk of ICU and hospital mortality.

Objective To explore the clinical efficacy of staged surgery for non-lactating mastitis. Methods A retrospective analysis was conducted on 317 patients with non-lactating mastitis admitted to our department from January 2015 to December 2020, all of whom underwent staged surgical treatment. The recovery time, recurrence rate, and breast appearance score were observed. Results The median follow-up time was 24(17，33) months, the recovery time was (25.5±17.9) days, and the recurrence rate was 4.4%. There were 96.2% of patients satisfied with the breast appearance. Conclusions Staged surgery for non-lactating mastitis can effectively shorten the course of the disease, protect the appearance of breast, and have good clinical efficacy.

End-stage renal disease (ESRD) patients undergoing outpatient hemodialysis (HD) and home peritoneal dialysis (PD) are high risk population of severe and critical types caused by SARS-CoV-2 infection. In order to improve the quality of diagnosis and treatment in dialysis patients with SARS-CoV-2 infection, we wrote this recommendation for primary care clinicians. During the epidemic period of SARS-CoV-2 infection, all patients should be instructed to strengthen self-management. Once the SARS-CoV-2 infection was found in dialysis patients, early stratified management should be carried out within 72 hours after the first positive nucleic acid or antigen test results, which includes early antiviral therapy, early recognition, and transferring severe patients from community or primary hospital to a referral hospital promptly. Guidance for dietary and sports rehabilitation after SARS-CoV-2 infection should also be started as soon as possible.

Considering that photodynamic therapy (PDT)-induced oxygen consumption and microvascular damage could exacerbate hypoxia to drive more glycolysis and angiogenesis, a novel approach to potentiate PDT and overcome the resistances of hypoxia is avidly needed. Herein, morpholine-modified PEGylated bilirubin was proposed to co-deliver chlorin e6, a photosensitizer, and diclofenac (Dc). In acidic milieu, the presence of morpholine could enable the nanocarriers to selectively accumulate in tumor cells, while PDT-generated reactive oxidative species (ROS) resulted in the collapse of bilirubin nanoparticles and rapid release of Dc. Combining with Dc showed a higher rate of apoptosis over PDT alone and simultaneously triggered a domino effect, including blocking the activity and expression of lactate dehydrogenase A (LDHA), interfering with lactate secretion, suppressing the activation of various angiogenic factors and thus obviating hypoxia-induced resistance-glycolysis and angiogenesis. In addition, inhibition of hypoxia-inducible factor-1α (HIF-1α) by Dc alleviated hypoxia-induced resistance. This study offered a sequentially responsive platform to achieve sufficient tumor enrichment, on-demand drug release and superior anti-tumor outcomes in vitro and in vivo.

Objective:To screen the neutralizing epitope of enterovirus 71 (EV71) and determine the specific minimum amino acid sequence that triggers immunity for providing a theoretical basis for the development of synthetic peptide vaccines.Methods:EV71 neutralizing antibody-specific binding clones were panned and sequenced using a phage display random 12-peptide library to obtain the key sequences of neutralizing epitopes. A series of peptides containing the key sequences with N-terminal acetylation (AC) and C-terminal linking to Keyhole limpet hemocyanin (KLH) were synthesized. Serum samples were collected after immunizing mice with the modified peptides. Then the immunogenicity of the peptides and the neutralizing activity of serum samples were analyzed by Western blot, ELISA and neutralization test.Results:After three rounds of panning, cloning and sequencing, KQEKDL was identified as the key motif. The serum samples collected from the mice immunized with the modified series of peptides containing key motifs had different degrees of binding ability to EV71 and VP1 protein. The serum samples of mice immunized the synthetic peptide containing only the minimum key motif (AC-KQEKDL-KLH) had the strongest response to the other three peptides and EV71 and the highest neutralizing titer.Conclusions:The EV71 neutralizing epitope was successfully screened using the phage display random peptide library. The key motif of KQEKDL might be the specific minimum amino acid sequence that triggered the immune system. This study provides a theoretical basis for better understanding the immune response mechanism, evaluating the immunogenicity of the antigens and further research and development of polypeptide vaccines.

Objective:To investigate the clinical characteristics and prognosis of patients with cutaneous intravascular large B-cell lymphoma (IVLBCL).Methods:The data of 30 cutaneous IVLBCL published between January 1989 and May 2019 in China were systematically reviewed. The clinical manifestation, biochemical and imaging characteristics and diagnostic features of patients were summarized, and then the survival of different groups was also analyzed.Results:The median onset age was 61.5 years old (25.0-83.0 years old), and there were 22 (73.3%) females. All 30 patients presented with cutaneous lesions. Initial symptoms showed cutaneous lesions in 16 (53.3%) patients; and B symptom, respiratory symptoms or central nervous system (CNS) occurred in 14 (46.7%) patients with late cutaneous lesions. Cutaneous lesions were heterogeneous, and 76.7% (23/30) lesions located in lower abdomen and proximal limbs. And 76.2% (16/21) were positive in image examination, and 78.3% (18/23) had two or more extranodal organs invasion. The median time from onset to visit was 2.5 months (0.4-24.0 months), and clinical misdiagnosis rate was 56.7%(17/30). All IVLBCL patients were confirmed by biopsy, including 6 cases (27.3%, 6/22) of bone marrow involvement, 1 case (3.3%) of hemophagocytic syndrome-associated variant, and 29 cases (96.7%) of classical variant. Finally, 81.8% (18/22) patients received anthracycline-based combined chemotherapy. Compared with non-chemotherapy group, the median OS time of chemotherapy group was prolonged [11.0 months (2.0-60.0 months) vs. 2.0 months (0.7-24.0 months), P = 0.002]. Patients with CNS symptoms had shorter median OS time compared with patients without CNS symptoms [2.0 months (0.7-6.0 months) vs. 11.0 months (1.0-60.0 months), P < 0.01]. The median OS time in the group of cutaneous lesions as initial symptom combined with other symptoms was longer than that in group of late cutaneous lesions and other symptoms as initial symptom [unreached (2.0-60.0 months) vs. 3.0 months (1.5-24.0 months), P = 0.032]. Conclusions:Cutaneous IVLBCL is a rare disease with atypical clinical characteristics in China. Prompt attention and biopsy in time will be helpful for early diagnosis. Accompanied with CNS symptoms suggests poor prognosis; and timely chemotherapy can improve the prognosis of the patients.

Objective:To investigate the treatment, safety and prognosis of advanced non-small cell lung cancer patients with leptomeningeal metastasis and performance status score more than 3.Methods:The clinical data of 6 NSCLC patients with leptomeningeal metastasis admitted to the People's Hospital of Wuhan University from November 2016 to September 2018 were analyzed retrospectively. The curative effect and adverse reactions were observed, and the prognosis was analyzed.Results:There were 5 females and 1 male among 6 patients. The median age was 57 years old (46-74 years old). All 6 patients were diagnosed as stage Ⅳ lung adenocarcinoma. There were 3 patients with epidermal growth factor receptor (EGFR) exon 21 mutation, 2 patients with exon 19 mutation and one with anaplastic lymphoma kinase (ALK) fusion mutation. The time window of leptomeningeal metastasis occurred after the progression of adenocarcinoma of lung: 3 cases was more than 12 months, the other 3 cases was less than 12 months, and the average was 20.3 month. Performance status score was more than 3 when leptomeningeal metastasis occurred. The brain magnetic resonance imaging of 6 patients showed linear enhancement of leptomeningeal, cancer cells were found in cerebrospinal fluid in one case, 4 cases were treated with a combination of bevacizumab and EGFR-tyrosine kinase inhibitor (EGFR-TKI), 1 case was treated with oral administration of EGFR-TKI, 1 case was treated with oral administration of EGFR-TKI combined with temozolomide. The median overall survival (mOS) was 9 months (2-13 months), and the median progression free survival was 6 months (2-11 months).Conclusion:Lung adenocarcinoma may be prone to leptomeningeal metastasis; for NSCLC patients with leptomeningeal metastasis and performance status score more than 3, a combination of EGFR-TKI and bevacizumab has good tolerance, high safety and considerable curative effect.