Gene therapy, harnessing the power of CRISPR-Cas9 and/or DNAzyme systems, stands as a pivotal approach in cancer therapy, enabling the meticulous manipulation of genes pivotal to tumorigenesis and immunity. However, the pursuit of precise gene therapy encounters formidable hurdles. Herein, a near-infrared upconversion theranostic nanomachine is devised and tailors for CRISPR-Cas9/DNAzyme systems mediate precise gene therapy. An ingenious logic DNAzyme system consists of Chain 1 (C1)/Chain 2 (C2) and endogenous lncRNA is designed. We employ manganese modified upconversion nanoparticles for carrying ultraviolet-responsive C1-PC linker-C2 (C₂P) chain and Cas9 ribonucleoprotein (RNP), with outermost coats with hyaluronic acid. Upon reaching tumor microenvironment (TME), the released Mn²⁺ ions orchestrate a trifecta: facilitating endosomal escape, activating cGAS-STING signaling, and enabling T1-magnetic resonance imaging. Under near-infrared irradiation, Cas9 RNP/C₂P complex dissociates, releasing Cas9 RNP into the nucleus to perform gene editing of Ptpn2, while C1/C2 chains self-assemble with endogenous lncRNA to form a functional DNAzyme system, targeting PD-L1 mRNA for gene silencing. This strategy remodels the TME by activating cGAS-STING signaling and dual immune checkpoints blockade, thus realizing tumor elimination. Our theranostic nanomachine armed with the CRISPR-Cas9/DNAzyme logic systems, represents a resourceful and promising strategy for advancing cancer systemic immunotherapy and precise gene therapy.

Objective To investigate the correlation between complement C3,C4,C-reactive protein(CRP),erythrocyte sedimentation rate(ESR)and disease activity of systemic lupus erythematosus(SLE).Methods A total of 69 SLE patients admitted to the hospital from January to December 2021 were selected as the SLE group,and 70 healthy individuals were selected as the control group during the same period.The changes of serum related indicators were compared between the SLE group and the control group.According to systemic lupus erythematosus disease activity index(SLEDAI),SLE patients were divided into mild active group(27 cases)and moderate and severe active group(42 cases).The changes of related indicators were compared between the two groups.Spearman correlation analysis was used to analyze the correlation between SLEDAI score and serum related indicators.Receiver operating characteristic(ROC)curve was used to ana-lyze the validity of diagnosis of disease severity.Results Compared with the control group,the levels of serum complement C3 and C4 in the SLE group were decreased,and the levels of CRP and ESR were increased,the differences were statistically significant(P＜0.05).Compared with the mild group,the levels of serum com-plement C3 and C4 in the moderate and severe group were decreased,and the levels of CRP and ESR were in-creased,and the differences were statistically significant(P＜0.05).The SLEDAI score of SLE patients was positively correlated with serum CRP and ESR levels(P＜0.05),and negatively correlated with serum com-plement C3 and C4 levels(P＜0.05).The area under the curve(AUC)of CRP,ESR,complement C3 and C4 were 0.716,0.875,0.872 and 0.856,respectively,the sensitivity were 59.52％,85.71％,78.57％and 78.79％,respectively,and the specificity were 79.83％,81.67％,86.79％and 86.54％,respectively.Conclu-sion Serum complement C3,C4,CRP and ESR are correlated with the progression of SLE,and play auxiliary roles in the disease activity stage of SLE.

Objective To investigate and analyze the current status and challenges of infant and toddler nutritional services in urban and rural medical facilities in Sichuan Province.Methods In 2022,a questionnaire survey was conducted to collect data on infant and toddler nutritional services,including feeding guidance,physical growth assessment,and micronutrient deficiency screening,as well as information on personnel and tools in medical facilities throughout Sichuan Province.The provision of nutritional services was analyzed and the urban-rural disparities were assessed.Results A total of 2206 medical facilities(29.1%from urban areas and 70.9%from rural areas)were investigated.Only 35.8%of medical facilities provided all three types of nutritional services.Specifically,the overall service provision rates were high for feeding guidance(94.6%)and physical growth assessment(85.0%),but lower for micronutrient deficiency screening(37.4%).Rural facilities exhibited significantly lower rates than their urban counterparts for both physical growth assessment and micronutrient deficiency screening(P＜0.05).The provision rates of feeding guidance ranged from 70.6%to 93.2%,with responsive feeding guidance being the least implemented(70.6%),particularly in rural areas compared to urban areas(P＜0.05).Rates for physical growth assessment and micronutrient deficiency screening ranged from 75.3%to 81.8%and 23.6%to 30.8%,respectively,both showing lower rates in rural settings compared to urban ones(P＜0.05).Nutrition service providers were predominantly nurses(52.3%)and clinical practitioners(43.4%).The availability of dietary assessment tools ranged from 7.7%to 15.9%,significantly lower in rural areas compared to urban areas(P＜0.001),while physical measurement tools were widely available at rates of 94.6%to 98.5%.Conclusion At present,the infant and toddler nutritional service provisions of medical facilities in Sichuan Province are incomplete,particularly so in the implementation of feeding guidance,physical growth assessment,and micronutrient deficiency screening.There is a notable shortage of personnel and necessary tools,with rural areas facing more significant challenges.Enhancing the overall capacity of infant and toddler nutritional services in Sichuan Province is essential,with specific attention needed for rural healthcare settings.

Objective:To compare the clinical value of stent assisted intestinal bypass and temporary loop ileostomy in laparoscopic low anterior resection of rectal cancer.Method:In this retrospective analysis, 57 patients undergoing laparoscopic low anterior resection for rectal cancer in the First Affiliated Hospital of Ningbo University from Jan 2020 to Jan 2022 were divided into intestinal bypass group (36 cases) and loop ileostomy group (21 cases).Result:There were no significant differences in postoperative GI function recovery and postoperative complication rate between the two groups (all P>0.05). The levels of albumin, prealbumin and hemoglobin in the intestinal bypass group were better than those in the ileostomy group when evaluated on 3rd months after operation [(40.5±2.3) g/L vs. (38.1±2.6)g/L、(26.4±2.7)mg/dl vs. (24.5±2.0)mg/dl、(137.6±5.9) g/L vs. (134.0±7.0) g/L, t=3.605、2.743、2.085, all P<0.05]. Hospital expenses of the intestinal bypass group was lower [(571 000±7 500) yuan vs. (69 300±9 100) yuan, t=-5.477, P<0.05]. Conclusion:Compared with traditional ileostomy, the stent assisted intestinal bypass reduces trauma with lower expenses and improves patients' status after laparoscopic low anterior resection for rectal cancer.

Neurogenesis decline in hippocampal dentate gyrus (DG) participates in stress-induced depressive-like behaviors, but the underlying mechanism remains poorly understood. Here, we observed low-expression of NOD-like receptor family pyrin domain containing 6 (NLRP6) in hippocampus of stress-stimulated mice, being consistent with high corticosterone level. NLRP6 was found to be abundantly expressed in neural stem cells (NSCs) of DG. Both Nlrp6 knockout (Nlrp6^-/-) and NSC-conditional Nlrp6 knockout (Nlrp6CKO) mice were susceptible to stress, being more likely to develop depressive-like behaviors. Interestingly, NLRP6 was required for NSC proliferation in sustaining hippocampal neurogenesis and reinforcing stress resilience during growing up. Nlrp6 deficiency promoted esophageal cancer-related gene 4 (ECRG4) expression and caused mitochondrial dysfunction. Corticosterone as a stress factor significantly down-regulated NLRP6 expression, damaged mitochondrial function and suppressed cell proliferation in NSCs, which were blocked by Nlrp6 overexpression. ECRG4 knockdown reversed corticosterone-induced NSC mitochondrial function and cell proliferation disorders. Pioglitazone, a well-known clinical drug, up-regulated NLRP6 expression to inhibit ECRG4 expression in its protection against corticosterone-induced NSC mitochondrial dysfunction and proliferation restriction. In conclusion, this study demonstrates that NLRP6 is essential to maintain mitochondrial homeostasis and proliferation in NSCs, and identifies NLRP6 as a promising therapeutic target for hippocampal neurogenesis decline linked to depression.

ObjectiveTo reveal the targets and molecular mechanisms of the action of Qiangxin Decoction （强心汤） for the treatment of chronic heart failure based on the combination of network pharmacology and molecular docking. MethodsThe active ingredients of Qiangxin Decoction were retrieved from TCMSP database， and the targets of chronic heart failure were screened by searching GeneCards， OMIM， TTD， PharmGkb， and DrugBank databases， and the intersections were taken to obtain the intersecting targets of Qiangxin Decoction for the treatment of chronic heart failure. STRING platform was used to construct the protein-protein interaction network （PPI）， Cytoscape 3.8.0 software was used to calculate the network topology to screen the core targets， and R 4.2.3 was used to construct the “active ingredient-target” network by analyzing the GO enrichment analysis and KEGG pathway enrichment analysis. AutoDock 1.5.7 was used for molecular docking to predict the binding performance of active ingredients and core targets. ResultsSeventy-five intersecting targets were identified for the treatment of chronic heart failure with Qiangxin Decoction， among which the core targets were estrogen receptor 1 （ESR1， degree value=7）， nuclear receptor coactivator 1 （NCOA1， degree value=8）， glucocorticoid receptor （NR3C1， degree value=7）， and nuclear receptor coactivator 2 （NCOA2， degree value=7）. GO enrichment analysis showed that the top 3 items with the smallest P value in molecular function were G protein-coupled amine receptor activity， postsynaptic neurotransmitter receptor activity， and neurotransmitter receptor activity （P＜0.01）； the top 3 items with the smallest P value in biological process were adenylyl cyclase-activated adrenergic receptor signaling pathway， adrenergic receptor signaling pathway， and adenylyl cyclase-regulated G protein-coupled receptor signaling pathway （P＜0.01）； the top 3 items with the smallest P values in cellular composition were components of the postsynaptic membrane， synaptic membrane， and presynaptic membrane （P＜0.01）. KEGG enrichment analysis showed that the top 5 key signaling pathways were neuroactive ligand-receptor interactions， calcium signaling pathway， dopaminergic synapses， cocaine addiction， and cyclic guanosine monophosphate-protein kinase G （cGMP-PKG） signaling pathway. The molecular docking results showed that lignans and isoflavones had lower binding energies and more structural stability with the four core targets （ESR1， NCOA1， NR3C1， NCOA2）. ConclusionThe treatment of chronic heart failure by Qiangxin Decoction was associated with neuroactive ligand-receptor interactions， calcium signaling pathway， dopaminergic synapses， chemoattractant-receptor activation， cGMP-PKG signaling pathway， lipids and atherosclerosis， and cAMP signaling pathway， and lignans and isoflavones may be the core active compounds in its treatment of chronic heart failure.

Objective: To analyze the phenotype and genotype of a Chinese pedigree with inherited dysfibrinogenaemia and investigate the molecular mechanism of the disease. Methods: Venous blood samples were collected from all family members, and routine coagulation tests were conducted. Functional fibrinogen in venous blood samples was measured by Clauss method, and the antigen level of fibrinogen in plasma was measured by immunoturbidimetry assay. All the exons and exon-intron boundaries of the three fibrinogen genes were analyzed by direct sequencing. Fibrinogen electrophoresis, fibrinogen clottability measurement, fibrin polymerisation measurement and electron microscopy scanning were also used to investigate the molecular characteristics and pathogenesis. Results: The proband had normal activated partial thromboplastin time, prothrombin time and plasma fibrinogen antigen, but prolonged thrombin time, prolonged reptilase time and reduced fibrinogen activity level, which were also found in his father. The sequencing results of the proband revealed heterozygous A1211G in the exon 2 of FGA gene originating from his father, which caused Arg19Gly missense mutation. The western-blot results showed that no abnormal bands of plasma fibrinogen were found in the proband and his father. Both thrombin-induced fibrin polymerisation and reptilase induced fibrin polymerisation were significantly impaired compared to normal control. Fibrinogen clottability measurement showed that only about 20.8% molecules of plasma fibrinogen in the proband were involved in the clot formation. Scanning electron microscopy revealed that the proband′s average fibre diameters were found to be significantly thicker than that of the control(P＜0.001), and the density was smaller than that of normal control. Conclusion The Arg19Gly mutation should be responsible for the proband′s dysfibrinogenaemia and the relevant clinical symptoms.

Objective To investigate the clinicopathological features and correlation between synchronous multiple early gastric cancer (SMEGC)and single early gastric cancer (EGC).Methods From January 2008 to December 2016,the clinical data of 994 patients with EGC who underwent open or laparoscopic gastrectomy surgery were collected from the electronic medical data base of the First Affiliated Hospital of Nanjing Medical University and Xuzhou No.1 People's Hospital.The data of patients including gender,age,tumor morphologys,tumor location,tumor size,histological type,depth of invasion,lymph nodes metastasis,lymphovascular metastasis,peripheral nerve invasion,and blood types were analyzed.T test and Chi square test were used for statistical analysis.Results Among 994 EGC patients,27 cases (2.7％) were SMEGC,and 967 cases (97.3％) were single EGC.The percentage of male and female of single EGC were 71.4％ (690/967) and 28.6％ (277/967),respectively;the percentage of male and female of SMEGC were 88.9％ (24/27) and 11.1％ (3/27),respectively,and there was statistically significant difference in the gender composition ratio between single EGC and SMEGC (x2 =3.975,P=0.046).The incidence of ulcer in single EGC and SMEGC were 50.6％ (489/ 967) and 29.6 ％ (8/27),respectively,and the difference in the incidence of ulcers between single EGC and SMEGC was statistically significant (x2 =4.653,P=0.031).There were no statistically significant differences between single EGC and SMEGC in gross morphology,depth of invasion,lymph nodes metastasis,lymphovascular metastasis,peripheral nerve invasion,tumor location,pathological type and blood types (all P＞0.05).In the SMEGC patients,the incidence of main lesions invading the mucosa was 48.1％ (13/27) and submucosa invasion was 51.9％ (14/27);and for minor lesions,the corresponding incidences were 77.8％ (21/27) and 22.2％ (6/27),respectively,and the difference was statistically significant (x2 =5.063,P＜0.05).There were no statistically significant differences between the main lesions and minor lesions in tumor size,pathological type,with or without ulcers,gross morphology and tumor location (all P＞0.05).Conclusions The main risk factors of SMEGC are male and no ulcerative lesions.The clinicopathological features are similar between main lesions and minor lesions in SMEGC.

Objective:To explore the feasibility and efficacy of short peptide enteral nutrition in preoperative bowel preparation for patients with gynecological laparoscopy.Methods:A total of 100 patients with gynecological laparoscopy were randomized into the trial group (n =50) and the control group (n =50) by using the random number table method.Patients in the study group received short peptide enteral nutrition for three days before operation without mechanical bowel preparation.Patients in the control group received traditional liguid diet and mechanical bowel preparation.The satisfaction of bowel preparation,postoperative anal exhaust time,hospitalization time and nutritionrelated parameters were compared between the two groups.Results:Doctor's satisfaction of bowel preparation in the study group was significantly higher than the control group (90％ vs 64％) (P ＜ 0.01).Postoperative anal exhaust time and hospitalization time were shorter in the control group (both P ＜ 0.05).The levels of postoperative albumin and hemoglobin in study group were significantly higher than the control group (bothP ＜ 0.05).Conclusion:Short peptide EN in preoperative bowel preparation of gynecological laparoscopy reveals a better efficacy and safety,a better recovery in postoperative intestinal function and nutritional status,and a shorter postoperative hospitalization time.It is well worth clinical promotion.

In this study, the rescue effect of receptor activator for nuclear factor-kappaB ligand (RANKL) on zoledronate acid (ZOL) induced inhibition of osteoclastogenesis and gene expression of NF-kappaB p50 and c-Jun was investigated. Mice calvarial osteoblasts (OBs) were harvested and co-cultured with RAW264.7 cells and the cells were divided into 4 groups and received treatment with ZOL and RANKL, either single or combined. The formation of multi-nucleated osteoclast (OC) was examined and gene expression of NF-kappaB p50 and c-Jun was detected. Group B (ZOL) showed least multi-nucleated OC and resorption lacunae among the 4 groups (P < 0.05 or P < 0.01) and it was followed by group C (ZOL+RANKL). Group D (RANKL) showed highest OC and resorption lacunae while it was similar to Group A (control) (P > 0.05). Gene expression of NF-kappaB p50 and c-Jun was the lowest in group B (P < 0.05 or P < 0.01) among the four groups and was significantly increased in group C when compared with group B (P < 0.05). Group A and D showed highest gene expression and they were similar to each other (P > 0.05). This study suggest that RANKL might partly rescue ZOL induced inhibition of osteoclastogenesis, and the effect of RANKL and ZOL on osteoclastogenesis may be mediated by NF-kappaB p50 and c-Jun.
Animals ; Bone Resorption ; drug therapy ; Cell Line ; Diphosphonates ; pharmacology ; Gene Expression ; Imidazoles ; pharmacology ; Mice ; NF-kappa B p50 Subunit ; metabolism ; Osteoblasts ; drug effects ; Osteoclasts ; drug effects ; Proto-Oncogene Proteins c-jun ; metabolism ; RANK Ligand ; pharmacology