ObjectiveTo investigate the role of Krüppel-like factor 4 （KLF4） in type 1 diabetic nephropathy （DN） and to elucidate its underlying mechanisms. MethodsSixteen male Sprague-Dawley （SD） rats were selected and randomly divided into control group and model group， with 8 rats in each group. Rats in model group were intraperitoneally injected with a single dose of 55 mg/kg streptozotocin （STZ） to establish a diabetic nephropathy （DN） model， while those in control group were injected with an equal volume of sodium citrate buffer at the same time. After successful model establishment， the serum levels of blood urea nitrogen （BUN） and serum creatinine （SCR） were determined. Hematoxylin-eosin （HE） staining was performed on renal tissues to observe pathological changes， and immunofluorescence staining was conducted to detect the expression of KLF4 in renal tissues. Lipid peroxidation levels were evaluated by measuring malondialdehyde （MDA）， Fe²⁺， and lipid peroxidation products in rat kidneys. A high glucose （HG）-induced cell injury model was established in HK-2 cells， with mitochondrial membrane potential assessed using 5，5'，6，6'-tetrachloro-1，1'，3，3'- tetraethylbenzimidazolylcarbocyanine iodide （JC-1） staining. Lipid peroxidation levels （MDA， Fe²⁺， and lipid peroxides） were measured in HK-2 cells.KLF4-overexpressing HK-2 cells were then constructed， followed by repeated JC-1， MDA， Fe²⁺， and lipid peroxidation assays. Western blot was performed to evaluate the expression of ferroptosis-related proteins including glutathione peroxidase 4 （GPX4）， nuclear factor erythroid 2-related factor 2 （NRF2）， and Kelch-like ECH-associated protein 1 （Keap1）， in renal tissues， HK-2 cells， and KLF4-overexpressing HK-2 cells. ResultsCompared with the control group， DN rats exhibited elevated serum BUN and SCR levels， glomerular hypertrophy， renal interstitial fibrosis， and decreased KLF4 expression. Additionally， MDA， Fe²⁺， and lipid peroxidation levels increased， indicating enhanced ferroptosis in renal tissues， accompanied by reduced GPX4 and NRF2 expression and elevated Keap1 levels. Similarly， HG-treated HK-2 cells showed decreased KLF4 expression， increased MDA， Fe²⁺ and lipid peroxidation， elevated ferroptosis， and dysregulated GPX4/NRF2/Keap1 expression. However， KLF4 overexpression reversed these alterations induced by high glucose treatment. ConclusionIn the renal tissues of type 1 diabetic rats， the expression of KLF4 decreases the level of ferroptosis increases， and KLF4 overexpression could alleviate HG-induced HK-2 cell injury.

Syndrome differentiation and treatment is an integral part of the traditional Chinese medicine （TCM） diagnostic and therapeutic system， whose development exhibits distinct stages. This paper systematically reviews the evolutionary trajectory of syndrome differentiation and treatment， from symptom-based treatment in Inner Canon of Yellow Emperor （《黄帝内经》）， to ZHANG Zhongjing's establishment of the disease-pulse-syndrome-treatment framework， through its application and development in the Ming and Qing dynasties， and finally to its recognition as a fundamental characteristic of TCM in modern times. However， the overemphasis on syndrome differentiation and treatment， coupled with a diminished focus on disease concepts， has led to its gradual alienation as the primary diagnostic and therapeutic model. The alienation mainly manifests as a tendency to prioritize syndrome over disease， resulting in the overgeneralization and limited application of the concept， and causing a lack of specificity in practice. This paper emphasizes that a correct understanding of syndrome differentiation and treatment is a necessary premise for the deve-lopment and improvement of the TCM diagnostic and therapeutic system. By integrating modern medical diagnosis to clarify disease targets and applying TCM thinking to extract common patterns of diseases， precise alignment between syndrome differentiation and treatment and modern clinical demands can be achieved， providing a reference for addressing the contemporary challenges of syndrome differentiation and treatment.

Objective:To investigate the effects of knockdown and overexpression of melanoma-associated antigen-A3(MAGE-A3)gene on the biological behavior and cisplatin resistance of ovarian cancer cells.Methods:The expression level of MAGE-A3 mRNA and protein in ovarian cancer cells was detected.The knockdown cell lines were constructed and divided into control group,no-load(NC)group and sh MAGE-A3 group.The overex-pression cell lines were divided into control group,no-load(NC)group and OE-MAGE-A3 group.The effects of knockdown and overexpression of MAGE-A3 gene on the proliferation,migration,invasion and apoptosis of tumor cells were compared.The expression of p53,Survivin,breast cancer susceptibility gene 1(BRCA1)and MAGE-A3 protein was detected.The high expression cell lines were divided into control group,cisplatin group,shMAGE-A3 group and cisplatin+shMAGE-A3 group.The low expression cell lines were divided into control group,cisplatin group,OE-MAGE-A3 group and cisplatin+OE-MAGE-A3 group.The cell proliferation,invasion ability of each group of cells,as well as their effects on the mRNA and protein expression level of each gene were compared.Results:The mRNA and protein expression of MAGE-A3 was high in SKOV3 cells and low in A2780 cells.Com-pared with the control group and NC group,the shMAGE-A3 group had lower OD450 value,migration rate and in-vasion numbers,higher apoptosis rate,lower expression of MAGE-A3 and Survivin protein,and higher expression of BRCA1 and p53 protein.The results of OE-MAGE-A3 group compared with control group and NC group were opposite to those of shMAGE-A3 group.Compared with the control group,cisplatin group and sh-MAGE-A3 group,the cisplatin+sh-MAGE-A3 group had lower OD450 value and invasion number higher levels of p53 and BRCA1 mRNA and protein expression,and lower levels of Survivin mRNA and protein.The OD450 value and in-vasion number of cells in cisplatin+OE-MAGE-A3 group were higher than those in cisplatin group,but lower than those in control group and OE-MAGE-A3 group.The mRNA and protein expression levels of p53 and BRCA1 were higher than those in the control group and the OE-MAGE-A3 group,but lower than those in the cisplatin group.The expression levels of Survivin mRNA and protein were lower than those in the control group and the OE-MAGE-A3 group,and higher than those in the cisplatin group.Conclusions:Knockdown of MAGE-A3 expression can effectively inhibit the proliferation,migration and invasion of ovarian cancer cells,promote apoptosis and en-hance cisplatin sensitivity.On the contrary,overexpression of MAGE-A3 increases cell proliferation,migration and invasion,and reduced apoptosis,thereby conferring cisplatin resistance.

Objective:To explore the relapse-related genes and their clinicopathological connections of diffuse large B cell lymphoma (DLBCL).Methods:Targeted panel sequencing was conducted on 32 eligible DLBCL samples; the patients were diagnosed, treated, and went into complete remission at the First Affiliated Hospital of Nanjing Medical University from January 2015 to December 2019, including 14 cases with recurrence (relapsed group) and 18 cases with long-term complete remission of over five years (remission group). Clinical and pathological data were further reviewed. Fisher′s exact test was employed to compare the differences in clinicopathological characteristics and mutation patterns between the two groups.Results:Among the 32 patients, there were 18 males and 14 females, with a male to female ratio of 1.3∶1.0 and a median age of 53 (45.5, 67.0) years. In the relapsed group, PIM1 (11/14), KMT2D (7/14), PRDM1 (6/14), MYD88 (6/14), DTX1 (6/14) emerged as the most frequently mutated genes. In the remission group, while recurrent PIM1, KMT2D and MYD88 mutations were also observed, the TP53 gene exhibited the highest mutation frequency (6/18). Compared to the remission group, relapsed group showed elevated mutation frequencies of PIM1 ( P=0.013) and FAT4 ( P=0.010), alongside a reduced incidence of TP53 mutations. In all 32 patients, DLBCL with CD79B, CCND3, DTX1, KMT2D and PRDM1 mutations demonstrated a propensity towards advanced clinicopathologic stage. Conclusions:Relapsed DLBCL has distinctive clinicopathological and genetic features. PIM1 and FAT4 may be served as potential biomarkers for screening relapsed DLBCL-NOS and as targets for novel therapeutic strategies.

Objective:To explore the relapse-related genes and their clinicopathological connections of diffuse large B cell lymphoma (DLBCL).Methods:Targeted panel sequencing was conducted on 32 eligible DLBCL samples; the patients were diagnosed, treated, and went into complete remission at the First Affiliated Hospital of Nanjing Medical University from January 2015 to December 2019, including 14 cases with recurrence (relapsed group) and 18 cases with long-term complete remission of over five years (remission group). Clinical and pathological data were further reviewed. Fisher′s exact test was employed to compare the differences in clinicopathological characteristics and mutation patterns between the two groups.Results:Among the 32 patients, there were 18 males and 14 females, with a male to female ratio of 1.3∶1.0 and a median age of 53 (45.5, 67.0) years. In the relapsed group, PIM1 (11/14), KMT2D (7/14), PRDM1 (6/14), MYD88 (6/14), DTX1 (6/14) emerged as the most frequently mutated genes. In the remission group, while recurrent PIM1, KMT2D and MYD88 mutations were also observed, the TP53 gene exhibited the highest mutation frequency (6/18). Compared to the remission group, relapsed group showed elevated mutation frequencies of PIM1 ( P=0.013) and FAT4 ( P=0.010), alongside a reduced incidence of TP53 mutations. In all 32 patients, DLBCL with CD79B, CCND3, DTX1, KMT2D and PRDM1 mutations demonstrated a propensity towards advanced clinicopathologic stage. Conclusions:Relapsed DLBCL has distinctive clinicopathological and genetic features. PIM1 and FAT4 may be served as potential biomarkers for screening relapsed DLBCL-NOS and as targets for novel therapeutic strategies.

Long non-coding RNAs(lncRNAs)regulate gene expression at the transcriptional,post-transcription-al,or epigenetic levels,and play crucial roles in tumor cell proliferation,invasion,apoptosis,and metabolism.With ongoing research,an increasing number of human microproteins encoded by lncRNAs have been discovered,although their complete functional characterization remains to be explored.This article reviews the research progress on the func-tions and partial mechanisms of the small integral membrane protein(SMIM)family encoded by lncRNAs in tumor ini-tiation and progression.

Long non-coding RNAs(lncRNAs)regulate gene expression at the transcriptional,post-transcription-al,or epigenetic levels,and play crucial roles in tumor cell proliferation,invasion,apoptosis,and metabolism.With ongoing research,an increasing number of human microproteins encoded by lncRNAs have been discovered,although their complete functional characterization remains to be explored.This article reviews the research progress on the func-tions and partial mechanisms of the small integral membrane protein(SMIM)family encoded by lncRNAs in tumor ini-tiation and progression.

Improving the nitrogen use efficiency (NUE) of Brassica napus is of significant importance for achieving the national goal of zero growth in chemical fertilizer application and ensuring the green development of the rapeseed industry. This study aims to explore the effects of the nitrate transporter gene BnaNRT1.5s on the nitrogen transport and NUE of B. napus, providing excellent genetic resources for the development of nitrogen-efficient B. napus varieties. The spatiotemporal expression of BnaA05.NRT1.5 as a key nitrogen responsive gene was profiled by qRT-PCR at different growth stages and for different tissue samples of B. napus 'Westar'. Subcellular localization was employed to examine its expression pattern in the cells. Additionally, CRISPR/Cas9 was used to create BnaNRT1.5s knockout lines, which were subjected to hydroponic experiments under high nitrogen (12.0 mmol/L) and low nitrogen (0.3 mmol/L) conditions. After the seedlings were cultivated for 21 days, root and shoot samples were collected for weighing, nitrogen content determination, xylem sap nitrate content assessment, and calculation of total nitrogen and NUE. The B. napus nitrate transporter BnaA05.NRT1.5 was localized to the cell membrane. During the seedling and early bolting stages, BnaA05.NRT1.5 was predominantly expressed in roots, while it was highly expressed in old leaves and mature silique skin during the reproductive stage. Compared with the wild type, the mutant BnaNRT1.5s showed significant increases in the dry weight and total nitrogen of seedlings under both high and low nitrogen conditions. Under low nitrogen conditions, NUE in the roots of BnaNRT1.5s significantly improved. Notably, under both high and low nitrogen conditions, the nitrate content in the shoots of BnaNRT1.5s decreased significantly, while that in the roots increased significantly, resulting in a significantly decreased shoot-to-root nitrate content ratio. BnaNRT1.5s is involved in regulating the transport of nitrate from the roots to the shoots, and its mutation enhances nitrogen absorption and utilization in B. napus seedlings, promoting seedling growth. This study not only provides references for understanding the physiological and molecular mechanisms by which BnaNRT1.5s regulates NUE but also offers valuable genetic resources for improving NUE in B. napus.
Brassica napus/genetics* ; Anion Transport Proteins/metabolism* ; Nitrogen/metabolism* ; Nitrate Transporters ; Plant Proteins/metabolism* ; Nitrates/metabolism* ; Gene Expression Regulation, Plant ; Biological Transport

By reconstructing the integrated Chinese and western medicine diagnostic and treatment system， the "Two Reconstructions" theoretical framework establishes a standardized pathway of "classification-staging-syndrome differentiation"， which improves the accuracy of disease identification and strengthens the capacity for full-course intervention； in addition， by reconstructing the modern materia medica system， it innovatively integrates the traditional properties and efficacy of Chinese herbal medicinals with modern pharmacological mechanisms， forming a "state-target co-regulation" precise medication model， and builds a dose-effect theoretical system for prescriptions and medicinals， thereby enhancing both the targeting accuracy and dosage precision of therapeutic interventions. The "Two Reconstructions" theorecitcal framework is a key strategy for enhancing clinical efficacy. It can precisely identify "states" and "targets" for directed intervention， shift the focus of prevention and treatment earlier to enable full-cycle management， establish standardized paradigms for reproducible and evaluable efficacy， and expand the scope of clinical practice to address conditions without typical syndromes and critical illnesses. As a systematic pathway for innovation in TCM， this theoretical framework provides valuable insights and references for promoting the high-quality development of integrative Chinese and western medicine.

Objective:To investigate the efficacy and safety of fospropofol disodium(FP)in the induction and maintenance of general anesthesia in the adult patients graded Ⅰ or Ⅱ by the American Society of Anesthesiologists(ASA)undergoing elective surgery,and to provide the theoretical basis for application of EP in the induction and maintenance of general anesthesia.Methods:Adult patients of ASA grade Ⅰ or Ⅱ undergoing elective surgery were selected with a total of 100 patients recruited sequentially according to the time of visit,and they were randomly divided into FP group(50 cases)and propofol group(50 cases).All patients were prepared preoperatively,and received a slow injection of midazolam(2 to 3 mg)and sufentanil(0.3 μg·kg-1),followed by induction of anaesthesia 1 to 2 min later.The patients in FP group were given FP(10.0-12.5 mg·kg-1)intravenously,and the patients in propofol group were given propofol(1.5-2.0 mg·kg-1)intravenously.After the Modified Obserational Assessment Alertness/Sedation(MOAA/S)score dropped to 1,muscle relaxant was administrated and the induction was completed.During the maintenance of anaesthesia,the patients in FP group received a continuous intravenous infusion of FP at a rate of 12.5-15.0 mg·kg-1·h-1,and the patients in propofol group received a continuous infusion of propofol at a starting rate of 6 mg·kg-1·h-1.The patients in two groups additionally received remifentanil(0.1-0.4 μg·kg-1·min-1)for co-analgesia,and the rate of administration was adjusted according to the patient's status.Systolic blood pressure(SBP),diastolic blood pressure(DBP),mean arterial pressure(MAP),heart rate(HR)and bispectral index(BIS)values of the patients in two groups were recorded at different time points:before induction(T1),immediately after tracheal intubation(T2),5 min after induction(T3),10 min after induction(T4),20 min after induction(T5),30 min after induction(T6),40 min after induction(T7)and at the end of the procedure(T8).The time to onset of sedation/anaesthesia(MOAA/S≤1),the time to eye opening,and the time to awakening(MOAA/S=5)of the patients in two groups were recorded.The lowest intraoperative SBP and BIS values and the time required of the patients in two groups were observed.The incidence of adverse reactions related to agitation,choking,nausea,vomiting and cardiovascular system or respiratory system were compared between two groups.Results:There were no statistically differences in the general informations and the duration of surgery of patients between two groups(P＞0.05).The induction time of the patients in FP group(2.39 min)was significantly longer than that in propofol group(0.70 min)(P＜0.05).In the recovery period of general anesthesia,the eye opening time and recovery time of the patients in FP group were significantly longer than those in propofol group(P＜0.05).There were no significant differences in MAP of the patients between two groups at different time points(P＞0.05).The HR at T4,T5,T6,and T7 time points of the patients in FP group were lower than those in propofol group(P＜0.05).The lowest value of BIS of the patients in FP group was significantly smaller than that in propofol group,and the time taken to reach the lowest value of BIS in FP group was significantly longer than that in propofol group(P＜0.05).The time taken to reach the lowest value of SBP of the patients in FP group was longer than that in propofol group(P＜0.05).However,the lowest value of SBP of the patients and the incidence of adverse reations of the patients in two groups showed no statistical differences(P＞0.05).Conclusion:Compared with propofol,FP injection is safe and effective in the induction and maintenance of general anesthesia in adult patients with ASA class Ⅰ or Ⅱ undergoing elective surgery,with a low incidence of adverse reactions,which is a new anesthesia option.