Objective To explore the impact of number of positive regional lymph nodes (nPRLN) in N1 stage on the prognosis of non-small cell lung cancer (NSCLC) patients. Methods Patients with TxN1M0 stage NSCLC who underwent lobectomy and mediastinal lymph node dissection from 2010 to 2015 were screened from SEER database (17 Regs, 2022nov sub). The optimal cutoff value of nPRLN was determined using X-tile software, and patients were divided into 2 groups according to the cutoff value: a nPRLN≤optimal cutoff group and a nPRLN>optimal cutoff group. The influence of confounding factors was minimized by propensity score matching (PSM) at a ratio of 1 : 1. Kaplan-Meier curves and Cox proportional hazards models were used to evaluate overall survival (OS) and lung cancer-specific survival (LCSS) of patients. Results A total of 1316 patients with TxN1M0 stage NSCLC were included, including 662 males and 654 females, with a median age of 67 (60, 73) years. The optimal cutoff value of nPRLN was 3, with 1165 patients in the nPRLN≤3 group and 151 patients in the nPRLN>3 group. After PSM, there were 138 patients in each group. Regardless of before or after PSM, OS and LCSS of patients in the nPRLN≤3 group were superior to those in the nPRLN>3 group (P<0.001). N1 stage nPRLN>3 was an independent prognostic risk factor for OS [HR=1.52, 95%CI (1.22, 1.89), P<0.001] and LCSS [HR=1.72, 95%CI (1.36, 2.18), P<0.001]. Conclusion N1 stage nPRLN>3 is an independent prognostic risk factor for NSCLC patients in TxN1M0 stage, which may provide new evidence for future revision of TNM staging N1 stage subclassification.

This paper summarizes Professor WANG Yaoxian's experience in treating diabetic kidney disease （DKD） from the perspective of spleen and stomach based on the "internal heat leading to concretions" theory. It is considered that internal heat leading to concretions constitutes the core pathogenesis of DKD， with the spleen and stomach serving as the source of internal heat； therefore， treatment should be based on regulating the spleen and stomach. In the early stage of DKD， dysfunction of the spleen and stomach leads to the initial generation of internal heat. Common syndrome patterns include gastrointestinal heat accumulation and constrained heat in the liver and stomach， for which modified Gegen Qinlian Decoction （葛根芩连汤） can be used to clear heat bind while modified Dachaihu Decoction （大柴胡汤） is used to clear stomach and soothe liver， respectively. In the middle stage of DKD， weakness of the spleen and stomach results in the initial formation of concretions and conglomerations. Common patterns include spleen deficiency with prevalence of dampness and deficiency of both the spleen and kidney. Treatment emphasizes strengthening the spleen and resolving dampness， raising yang and boosting the stomach with modified Shengyang Yiwei Decoction （升阳益胃汤）， or supplementing spleen and boosting kidney， dissipating bind and dispe-ring concretions with modified Shenqi Dihuang Decoction （参芪地黄汤）， respectively. In the late stage of DKD， it is characterized by spleen and stomach depletion， and rampant accumulation of turbidity and toxin， and the common syndrome patterns are damp-turbidity obstruction in the middle jiao （焦） and spleen-kidney yang deficiency. Treatment aims to remove turbidity and harmonize the stomach， or to warm the kidney and strengthen the spleen while elimina-ting turbidity， using modified Dahuang Gancao Decoction（大黄甘草汤） and Jupi Zhuru Decoction （橘皮竹茹汤） or modified Baoyuan Decoction （保元汤） and Lizhong Decoction （理中汤）， respectively. In clinical practice， appropriate formulas and medications are flexibly selected according to specific syndromes.

ObjectiveTo evaluate the effects of Qingre Huoxue prescription combined with methotrexate （MTX） and folic acid （FA） on homocysteine （Hcy） levels and disease activity in patients with rheumatoid arthritis （RA） complicated by hyperhomocysteinemia （HHcy）. MethodsOn the basis of a multicenter real-world cohort from the China Registry of Traditional Chinese Medicine for Rheumatism （CERTAIN） platform，408 patients with RA complicated by HHcy were selected and included in this study. Treatment with Qingre Huoxue prescription was used as the exposure factor. According to whether Qingre Huoxue prescription was combined with MTX and FA，patients were assigned into an exposure group of 281 cases and a control group of 127 cases. Follow-ups were conducted every three months，with the primary outcome indicator being the achievement rate of target Hcy level. A generalized estimating equation model was adopted to analyze the dynamic changes in Hcy level under different medication regimens. Kaplan-Meier analysis was performed to evaluate the cumulative achievement rate of target Hcy level in the two groups，and Cox regression analysis was conducted to explore the predictive factors affecting the achievement rate of target Hcy level. Changes in erythrocyte sedimentation rate （ESR），C-reactive protein （CRP），28-joint disease activity score （DAS 28），and Health Assessment Questionnaire-Disability Index （HAQ-DI） scores before and after the first combined treatment with Qingre Huoxue prescription were compared between the two groups. ResultsThe Hcy level in the exposure group showed a gradual decreasing trend over time，while the decrease in Hcy level was not significant in the control group. At the 12-month follow-up，the achievement rate of target Hcy level in the exposure group （124/281，44.13%） was higher than that in the control group （23/127，18.11%，P<0.01）. Multivariable Cox regression analysis showed that treatment with Qingre Huoxue prescription ［hazard ratio（HR）=3.013，95% confidence interval （CI） 2.188-4.149， P<0.001］， absence of comorbidities （HR=2.035， 95%CI 1.352-3.061， P<0.001）， and baseline Hcy <15 μmol·L^-1 （HR=1.810， 95%CI 1.390-2.357， P<0.001） were significantly associated with an increased likelihood of achieving target Hcy levels within 6 months. In contrast， higher PGA scores were associated with a reduced likelihood of achieving target Hcy levels （HR=0.991， 95% CI 0.986-0.997， P=0.003）. After treatment，the exposure group demonstrated reductions in ESR，CRP，and DAS 28 scores （P<0.05） and had lower ESR，CRP and DAS 28 scores than the control group （P<0.05）. ConclusionEven after taking FA supplements，some RA patients undergoing MTX treatment still have high Hcy levels. When being used in conjunction with MTX and FA，Qingre Huoxue prescription can effectively lower the Hcy level and disease activity in RA patients with HHcy，outperforming Western treatment alone.

ObjectiveTo evaluate the effects of Qingre Huoxue prescription combined with methotrexate （MTX） and folic acid （FA） on homocysteine （Hcy） levels and disease activity in patients with rheumatoid arthritis （RA） complicated by hyperhomocysteinemia （HHcy）. MethodsOn the basis of a multicenter real-world cohort from the China Registry of Traditional Chinese Medicine for Rheumatism （CERTAIN） platform，408 patients with RA complicated by HHcy were selected and included in this study. Treatment with Qingre Huoxue prescription was used as the exposure factor. According to whether Qingre Huoxue prescription was combined with MTX and FA，patients were assigned into an exposure group of 281 cases and a control group of 127 cases. Follow-ups were conducted every three months，with the primary outcome indicator being the achievement rate of target Hcy level. A generalized estimating equation model was adopted to analyze the dynamic changes in Hcy level under different medication regimens. Kaplan-Meier analysis was performed to evaluate the cumulative achievement rate of target Hcy level in the two groups，and Cox regression analysis was conducted to explore the predictive factors affecting the achievement rate of target Hcy level. Changes in erythrocyte sedimentation rate （ESR），C-reactive protein （CRP），28-joint disease activity score （DAS 28），and Health Assessment Questionnaire-Disability Index （HAQ-DI） scores before and after the first combined treatment with Qingre Huoxue prescription were compared between the two groups. ResultsThe Hcy level in the exposure group showed a gradual decreasing trend over time，while the decrease in Hcy level was not significant in the control group. At the 12-month follow-up，the achievement rate of target Hcy level in the exposure group （124/281，44.13%） was higher than that in the control group （23/127，18.11%，P<0.01）. Multivariable Cox regression analysis showed that treatment with Qingre Huoxue prescription ［hazard ratio（HR）=3.013，95% confidence interval （CI） 2.188-4.149， P<0.001］， absence of comorbidities （HR=2.035， 95%CI 1.352-3.061， P<0.001）， and baseline Hcy <15 μmol·L^-1 （HR=1.810， 95%CI 1.390-2.357， P<0.001） were significantly associated with an increased likelihood of achieving target Hcy levels within 6 months. In contrast， higher PGA scores were associated with a reduced likelihood of achieving target Hcy levels （HR=0.991， 95% CI 0.986-0.997， P=0.003）. After treatment，the exposure group demonstrated reductions in ESR，CRP，and DAS 28 scores （P<0.05） and had lower ESR，CRP and DAS 28 scores than the control group （P<0.05）. ConclusionEven after taking FA supplements，some RA patients undergoing MTX treatment still have high Hcy levels. When being used in conjunction with MTX and FA，Qingre Huoxue prescription can effectively lower the Hcy level and disease activity in RA patients with HHcy，outperforming Western treatment alone.

ObjectiveTo investigate the characteristics of mitochondrial translational initiation factor 2 （MTIF2） gene methylation and its association with the development and progression of hepatocellular carcinoma （HCC）. MethodsMethSurv and EWAS Data Hub were used to perform the standardized analysis and the cluster analysis of MTIF2 methylation samples， including survival curve analysis， methylation signature analysis， the association of tumor signaling pathways， and a comparative analysis based on pan-cancer database. The independent-samples t test was used for comparison between two groups； a one-way analysis of variance was used for comparison between multiple groups， and the least significant difference t-test was used for further comparison between two groups. The Cox proportional hazards model was used to perform the univariate and multivariate survival analyses of methylation level at the CpG site. The Kaplan-Meier method was used to investigate the survival differences between the patients with low methylation level and those with high methylation level， and the Log-likelihood ratio method was used for survival difference analysis. ResultsGlobal clustering of MTIF2 methylation showed that there was no significant difference in MTIF2 gene methylation level between different races， ethnicities， BMI levels， and ages. The Kaplan-Meier survival curve analysis showed that the patients with N-Shore hypermethylation of the MTIF2 gene had a significantly better prognosis than those with hypomethylation （hazard ratio ［HR］=0.492， P<0.001）， while there was no significant difference in survival rate between the patients with different CpG island and S-Shore methylation levels （P>0.05）. The methylation profile of the MTIF2 gene based on different ages， sexes， BMI levels， races， ethnicities， and clinical stages showed that the N-Shore and CpG island methylation levels of the MTIF2 gene decreased with the increase in age， and the Caucasian population had significantly lower N-Shore methylation levels of the MTIF2 gene than the Asian population （P<0.05）； the patients with clinical stage Ⅳ had significantly lower N-Shore and CpG island methylation levels of the MTIF2 gene than those with stage Ⅰ/Ⅱ （P<0.05）. Clinical validation showed that the patients with stage Ⅲ/Ⅳ HCC had a significantly lower methylation level of the MTIF2 gene than those with stage Ⅰ/Ⅱ HCC and the normal population （P<0.05）. ConclusionN-Shore hypomethylation of the MTIF2 gene is a risk factor for the development and progression of HCC.

Mid-to-late stage ankle arthritis is a chronic degenerative disease that is extremely common in clinical practice.It is characterized by significant cartilage degeneration and subchondral bone sclerosis,accompanied by the formation of osteophytes around the joint,often leading to joint deformity.This condition causes severe pain in the patients during walking,severely restricts their activities,and affects their qualities of life.In recent years,with the continuous improvement of medical standards,the treatment methods for mid-to-late stage ankle arthritis have shown a diversified development trend.Non-surgical treatments primarily include activity restriction,orthotic devices,oral non-steroidal anti-inflammatory drugs(NSAIDs),and intra-articular injections of the talocrural joint.The surgical treatments primarily include joint distraction arthroplasty,periacetabular osteotomy,total ankle arthroplasty,and ankle arthrodesis.Tissue engineering therapy,as an emerging method,has also received considerable attention.This article systematically reviewed the selection principles and research progress of various treatment options for mid-to-late stage ankle arthritis,including traditional treatments,non-surgical treatments,surgical treatments,and tissue engineering treatments.By deeply analyzing the basic principles and advantages and disadvantages of each treatment method,and combining the latest research findings on clinical outcomes,a scientific and comprehensive clinical decision-making reference system was constructed to provide clearer and more comprehensive treatment choices for both doctors and patients,thereby effectively improving treatment outcomes and enhancing the quality of life for the patients.

To analyze the standard establishment approach and compilation rationale for metallic pharmaceutical packaging standard development in the 2025 edition of the Pharmacopeia of the People's Republic of China.This article systematically explained the background and process of establishing the guiding principles for metallic materials and containers used in pharmaceutical packaging in the Chinese Pharmacopoeia through basic information,relevant domestic and international standards,the establishment of key quality attributes of metallic pharmaceutical packaging materials,and the construction of metallic pharmaceutical packaging material standards.The newly established guidelines,the Pharmacopeia of the People's Republic of China 9625,prioritized product critical quality attributes(CQAs)and real-world applicability.This dual emphasis on rigidity and adaptability enhances drug safety,meets the regulatory requirements,and promotes the globalization and scientific advancement of China's pharmaceutical packaging industry.

Lung cancer poses a serious threat to global public health security.Chemotherapy,as the main strategy for cancer treatment,faces challenges such as high toxicity and drug resistance.Anticancer peptides have the potential of being developed into new anticancer drugs due to their advantages of broad-spectrum anticancer activity,rapid action,and difficulty in generating drug resistance,but they also face shortcomings such as weak activity and strong toxic side effects.The weakly acidic microenvironment of tumors(pH 6.5-6.8)provides a good idea for the design of anticancer peptides of high-efficiency and low-toxicity.Previously,we designed the acid-sensitive antibacterial peptide pHly-1 using the wolf spider(Lycosa singoriensis)toxin Lycosin-Ⅰ as a template.In this study,we found that pHly-1 also had acid-sensitive anticancer activity.Further alanine scanning analysis of pHly-1 was carried out,and we ob-tained a mutant pHTP-2 with better acid sensitivity,whose IC50(half maximal inhibitory concentration)against A549 cells was 15.68 μmol/L at pH 6.6 and was greater than 100 μmol/L at pH 7.4.At pH 6.6,pHTP-2 could act on various lung cancer cell lines and induce the death of A549 cells by rapid ly-sis;at pH 7.4,500 μmol/L pHTP-2 had weak toxicity to red blood cells(the hemolysis rate was ap-proximately 38％)and primary myocardial cells(the inhibition rate was 49.7％,with P＜0.05).Analy-sis of its charge,particle size,morphology,and secondary structure showed that at pH 6.6,the histidine in the sequence of pHTP-2 was protonated,increasing the positive charge(P＜0.01),decreasing the hy-drated particle size(P＜0.05)and forming an α-helical structure to induce membrane lysis of A549 cells.At pH 7.4,it was deprotonated,the positive charge decreases,a β-sheet structure was formed and self-aggregation occurred,limiting its effect on the A549 cell membrane and showing weak activity.In summary,pHTP-2 could respond to the weakly acidic microenvironment of tumors to exert selective cyto-toxic activity,effectively overcoming the shortcomings of anticancer peptides such as low efficiency and high toxicity.Our findings suggest that it is a high-quality lead molecule for anticancer drugs.

Objective To provide a comprehensive review of the modeling method of the empty bottle stimulation(EBS)anxiety model,including commonly used experimental animal strains and genders,animal grouping,modeling procedures,modeling duration,primary behavioral evaluation method,and the underlying pathological mechanisms.This aims to offer a reference for the application of the EBS anxiety model in anxiety disorder research.Methods Searches were conducted in databases such as CNKI and PubMed to collect all literature related to the EBS anxiety model,which were then systematically summarized and organized.Results(1)Male adult SD or Wistar rats are predominantly used as experimental animals;(2)The optimal modeling period is 2 weeks;(3)Behavioral evaluations primarily utilize the open field test,elevated plus maze test,and light-dark box test;(4)Pathological mechanisms involve abnormal neurotransmitter metabolism in brain regions such as the hippocampus,prefrontal cortex,and amygdala.Conclusions The EBS anxiety model exhibits an anxiety-like behavioral phenotype and associated neurobiological mechanisms,validating its utility as an animal model for the study of anxiety disorders.However,further exploration and refinement are required for its standardized construction protocol and the understanding of its mechanistic underpinnings.

Tailored lipid nanoparticles (LNPs)-mediated small interfering RNA (siRNA) nanomedicines show promise in treating liver disease, such as acute liver injury (ALI) and non-alcoholic steatohepatitis (NASH). However, constructing LNPs that address biosafety concerns, ensure efficient delivery, and target specific hepatic subcellular fractions has been challenging. To evade above obstacles, we develop three novel self-degradable "gemini-like" ionizable lipids (SS-MA, SS-DC, SS-MH) by incorporating disulfide bonds and modifying the length of ester bond and tertiary amino head. Our findings reveal that the disulfide-bond-bridged LNPs exhibit reduction-responsive drug release, improving both biosafety and siRNA delivery efficiency. Furthermore, the distance of ester bond and tertiary amino head significantly influences the LNPs' pK _a, thereby affecting endosomal escape, hemolytic efficiency, absorption capacity of ApoE, uptake efficiency of hepatocytes and liver accumulation. We also develop the modified-mannose LNPs (M-LNP) to target liver macrophages specifically. The optimized M-MH_LNP@TNFα exhibits potential in preventing ALI by decreasing tumor necrosis factor α (TNFα) levels in the macrophages, while MH_LNP@DGAT2 could treat NASH by selectively degrading diacylglycerol O-acyltransferase 2 (DGAT2) in the hepatocytes. Our findings provide new insights into developing novel highly effective and low-toxic "gemini-like" ionizable lipids for constructing LNPs, potentially achieving more effective treatment for hepatic diseases.