Objective: To explore the effects of Cldn14 gene knockout on renal metabolism and stone formation in rats，so as to provide reference for research in the field of urinary calium metabolism and stone formation. Methods: Cldn14 gene knockout homozygous rats and wild-type rats of the same age were randomly divided into 4 groups：wild-type control (WC) group，wild-type ethylene glycol (WE) group，gene knockout control (KC) group and gene knockout ethylene glycol (KE) group，with 10 rats in each group.The WE and KE groups were induced with ethylene glycol + ammonium chloride to form kidney stones，while the WC and KC groups received normal saline gavage.After 4 weeks of standard maintenance feeding，the urine samples were collected to detect the venous blood.The kidneys were collected for HE，Pizzolatto's staining and transmission electron microscopy.The protein in renal tissues was extracted to detect the expressions of Claudin16 and Claudin19. Results: Crystal deposition was observed in the renal tubular lumen of the WE and the KE groups，and more crystals were detected in the KE group.The WE group had a large number of intracytoplasmic black crystalline inclusions observed in renal tubular epithelial cells under transmission electron microscope，followed by the KE and KC groups.Compared with WC and WE groups，KC and KE groups had significantly decreased serum calcium and magnesium levels but significantly increased urinary calcium level.In addition，the urinary calcium level was higher in the WE group than in the WC group and higher in the KE group than in the KC group.The KE group had lower level of Claudin16，but there was no significant difference in the level of Claudin19 among the 4 groups(P>0.05). Conclusion: Knockout of Cldn14 gene alone cannot effectively reduce urinary calcium excretion or reduce the risk of stone formation in rats，which may be related to the decrease of Claudin16 level.

Probiotics are natural systems bridging synthetic biology, physical biotechnology, and immunology, initiating innate and adaptive anti-tumor immune activity. We previously constructed an all-in-one engineered food-grade probiotic Lactococcus lactis (FOLactis) which could boost the crosstalk among different immune cells such as dendritic cells (DCs), natural killer cells, and T cells. Herein, considering the limited clinical efficacy of naked personalized neoantigen peptide vaccines, we decorate FOLactis with tumor antigens by employing a Plug-and-Display system comprising membrane-inserted peptides. Intranodal injection of FOLactis coated with neoantigen peptides (Ag-FOLactis) induces robust DCs presentation and neoantigen-specific cellular immunity. Notably, Ag-FOLactis not only triggers a 45-fold rise in the quantity of locally reactive neoantigen-specific T cells but also induces epitope spreading in both subcutaneous and metastatic tumor-bearing models, leading to potent inhibition of tumor growth. These findings imply that Ag-FOLactis represents a powerful platform to rapidly and easily display antigens, facilitating the development of a bio-activated platform for personalized therapy.

Objective To construct an intelligent postoperative rehabilitation management platform and evaluate its effect in patients with head and neck cancer.Methods The postoperative intelligent rehabilitation management platform of head and neck cancer patients was divided into 5 layers from bottom to top,including the underlying environment,platform support,core database,analysis algorithm and application platform.Through the construction of head and neck tumor postoperative complications knowledge base and risk prediction model and wearable device auxiliary precision prediction risk,the head and neck cancer patients with postoperative intelligent rehabilitation management was realized.A historical controlled study was conducted and 90 participants with head and neck cancer were selected who underwent the surgical treatment in a hospital in Hangzhou,Zhejiang Province.The experimental group(45 participants admitted from February to July)received the intervention by the intelligent rehabilitation management platform,while the control group(45 patients from August to December)received the routine rehabilitation management.The outcome measures including NRS scores at 12 h,24 h,48 h and 72 h after surgery,incidence of postoperative complications,postoperative length of hospital stay,and patient satisfaction were compared between the 2 groups.Results The scores of incidences of complications,postoperative hospital stay,and NRS scores at 12 h,24 h,48 h and 72 h after surgery in the experimental group were lower than those in the control group(P＜0.05),while the scores of patient satisfaction in the experimental group were higher than those of the control group(P＜0.05).Conclusion The intelligent postoperative rehabilitation management platform can achieve accurate prediction of postoperative complications,reduce postoperative complications of head and neck tumors,timely intervene to relieve patients'pain and discomfort,shorten hospital stays,improve nursing work efficiency and improve patient satisfaction.

Objective:To explore the effect of combining transcranial magnetic stimulation (rTMS) with acupuncture in the treatment of patients with vascular cognitive impairment (VCI).Methods:A total of 40 VCI patients were randomly assigned to either the control group or the experimental group ( n=20 per group) using a random number table. Both groups received routine treatment and rTMS, while the experimental group was additionally provided with acupuncture for four weeks. The rTMS was at 10Hz at 90% of the motor threshold. Before and after the treatment, both groups′ cognition was evaluated using the Montreal Cognitive Assessment (MoCA) and the Mini-Mental State Examination (MMSE). Ability in the activities of daily living (ADL) was quantified using the modified Barthel Index (MBI). Serum levels of interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-α) were measured using enzyme-linked immunosorbent assays. Results:Significant improvement was observed in both groups′ average MoCA, MMSE, and MBI scores, along with reductions in peripheral serum IL-6 and TNF-α levels. The experimental group exhibited significantly higher MoCA subdomain scores in the visuospatial and executive function, attention, and delayed recall subdomains, as well as a higher total MoCA score compared to the control group. The average MMSE and MBI scores were significantly higher in the experimental group than the control group, and that group′s serum IL-6 and TNF-α levels were significantly lower, on average. Pearson correlation analysis demonstrated a positive correlation between the improvements in MoCA and MMSE scores and the reductions in IL-6 and TNF-α levels in both groups.Conclusions:rTMS combined with acupuncture effectively improves cognition and ADL ability among VCI patients. The underlying mechanism may be associated with the reduction of neuroinflammatory factors IL-6 and TNF-α.

Objective:To explore the effect of combining transcranial magnetic stimulation (rTMS) with acupuncture in the treatment of patients with vascular cognitive impairment (VCI).Methods:A total of 40 VCI patients were randomly assigned to either the control group or the experimental group ( n=20 per group) using a random number table. Both groups received routine treatment and rTMS, while the experimental group was additionally provided with acupuncture for four weeks. The rTMS was at 10Hz at 90% of the motor threshold. Before and after the treatment, both groups′ cognition was evaluated using the Montreal Cognitive Assessment (MoCA) and the Mini-Mental State Examination (MMSE). Ability in the activities of daily living (ADL) was quantified using the modified Barthel Index (MBI). Serum levels of interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-α) were measured using enzyme-linked immunosorbent assays. Results:Significant improvement was observed in both groups′ average MoCA, MMSE, and MBI scores, along with reductions in peripheral serum IL-6 and TNF-α levels. The experimental group exhibited significantly higher MoCA subdomain scores in the visuospatial and executive function, attention, and delayed recall subdomains, as well as a higher total MoCA score compared to the control group. The average MMSE and MBI scores were significantly higher in the experimental group than the control group, and that group′s serum IL-6 and TNF-α levels were significantly lower, on average. Pearson correlation analysis demonstrated a positive correlation between the improvements in MoCA and MMSE scores and the reductions in IL-6 and TNF-α levels in both groups.Conclusions:rTMS combined with acupuncture effectively improves cognition and ADL ability among VCI patients. The underlying mechanism may be associated with the reduction of neuroinflammatory factors IL-6 and TNF-α.

Objective To construct an intelligent postoperative rehabilitation management platform and evaluate its effect in patients with head and neck cancer.Methods The postoperative intelligent rehabilitation management platform of head and neck cancer patients was divided into 5 layers from bottom to top,including the underlying environment,platform support,core database,analysis algorithm and application platform.Through the construction of head and neck tumor postoperative complications knowledge base and risk prediction model and wearable device auxiliary precision prediction risk,the head and neck cancer patients with postoperative intelligent rehabilitation management was realized.A historical controlled study was conducted and 90 participants with head and neck cancer were selected who underwent the surgical treatment in a hospital in Hangzhou,Zhejiang Province.The experimental group(45 participants admitted from February to July)received the intervention by the intelligent rehabilitation management platform,while the control group(45 patients from August to December)received the routine rehabilitation management.The outcome measures including NRS scores at 12 h,24 h,48 h and 72 h after surgery,incidence of postoperative complications,postoperative length of hospital stay,and patient satisfaction were compared between the 2 groups.Results The scores of incidences of complications,postoperative hospital stay,and NRS scores at 12 h,24 h,48 h and 72 h after surgery in the experimental group were lower than those in the control group(P＜0.05),while the scores of patient satisfaction in the experimental group were higher than those of the control group(P＜0.05).Conclusion The intelligent postoperative rehabilitation management platform can achieve accurate prediction of postoperative complications,reduce postoperative complications of head and neck tumors,timely intervene to relieve patients'pain and discomfort,shorten hospital stays,improve nursing work efficiency and improve patient satisfaction.

Objective:To construct a prediction model of inadvertent perioperative hypothermia in patients under general anesthesia , and to apply to clinic to verify its performance.Methods:The data of 19 068 surgical patients in a Grade Ⅲ Class A hospital in Zhejiang Province from January 2016 to September 2020 were included. The model was constructed by using artificial intelligence technology based on deep learning, and the prediction effect of the model was tested by using the area under the subject operating characteristic curve and decision curve. Totally 2 157 surgical patients were included from October 2020 to March 2021 to test the prediction accuracy of the model.Results:The incidence of hypothermia was 13.89% (2 649/19 068) in the modeling group and 14.18% (306/2 157) in the validation group. The area under the subject operating characteristic curve of the prediction model was 0.724 (95% CI: 0.707-0.741), the sensitivity was 0.516, the specificity was 0.823, the cut-off value was 0.175, and the accuracy of practical application was 79.54%. Conclusions:This model can stably predict the incidence of perioperative inadvertent hypothermia in patients under general anesthesia, and provide reference for clinical prevention of inadvertent perioperative hypothermia.

Objective::Regarding the secondary prevention of cardiovascular disease (CVD), there is great interest in preventing recurrent cardiovascular events (RCVEs). The prognostic importance of liver fibrosis scores (LFSs) has previously been reported in various CVDs. We hypothesized that LFSs might also be useful predictors for RCVEs in patients with prior cardiovascular events (CVEs). Herein, we aimed to evaluate the associations of LFSs with RCVEs in a large, real-world cohort of coronary artery disease (CAD) patients with a prior CVE.Methods::In this multicenter prospective study, 6527 consecutive patients with angiography-diagnosed CAD who had experienced a prior CVE (acute coronary syndrome, stroke, percutaneous coronary intervention, or coronary artery bypass grafting) were enrolled. LFSs were computed according to the published formulas: non-alcoholic fatty liver disease fibrosis score (NFS) includes age, body mass index (BMI), impaired fasting glycemia or diabetes mellitus (DM), aspartate aminotransferase (AST)/alanine aminotransferase (ALT) ratio, platelets, and albumin; fibrosis-4 (FIB-4) includes age, AST, ALT, and platelets; Forns score includes age, gamma-glutamyltransferase (GGT), and platelets; BARD includes BMI, AST/ALT ratio, and DM; GGT/platelet ratio includes GGT and platelets; AST/ALT ratio includes AST and ALT; and AST/platelet ratio index includes AST and platelets. The originally reported cutoffs were used for the categorization of low-, intermediate-, and high-score subgroups. All patients were followed up for the occurrence of RCVEs (comprising cardiovascular death, non-fatal myocardial infarction, and stroke). Cox and Poisson regression analyses were used to assess the relationship of baseline LFSs with the risk of RCVE.Results::During a mean follow-up of (54.67 ± 18.80) months, 532 (8.2%) RCVEs were recorded. Intermediate and high NFS, FIB-4, Forns, and BARD scores were independently associated with an increased risk of RCVE (hazard ratios ranging from 1.42 to 1.75 for intermediate scores and 1.35 to 2.52 for high scores). In the subgroup analyses of sex, age, BMI, DM, and hypertension status, the increased risk of RCVEs with high LFSs (NFS, FIB-4, Forns, and BARD) was maintained across the different subgroups (all P < 0.05). Conclusion::This study showed that LFSs are indeed independently associated with RCVEs, suggesting that LFSs may be used as novel tools for risk stratification in CAD patients with a prior CVE.

Objective::Regarding the secondary prevention of cardiovascular disease (CVD), there is great interest in preventing recurrent cardiovascular events (RCVEs). The prognostic importance of liver fibrosis scores (LFSs) has previously been reported in various CVDs. We hypothesized that LFSs might also be useful predictors for RCVEs in patients with prior cardiovascular events (CVEs). Herein, we aimed to evaluate the associations of LFSs with RCVEs in a large, real-world cohort of coronary artery disease (CAD) patients with a prior CVE.Methods::In this multicenter prospective study, 6527 consecutive patients with angiography-diagnosed CAD who had experienced a prior CVE (acute coronary syndrome, stroke, percutaneous coronary intervention, or coronary artery bypass grafting) were enrolled. LFSs were computed according to the published formulas: non-alcoholic fatty liver disease fibrosis score (NFS) includes age, body mass index (BMI), impaired fasting glycemia or diabetes mellitus (DM), aspartate aminotransferase (AST)/alanine aminotransferase (ALT) ratio, platelets, and albumin; fibrosis-4 (FIB-4) includes age, AST, ALT, and platelets; Forns score includes age, gamma-glutamyltransferase (GGT), and platelets; BARD includes BMI, AST/ALT ratio, and DM; GGT/platelet ratio includes GGT and platelets; AST/ALT ratio includes AST and ALT; and AST/platelet ratio index includes AST and platelets. The originally reported cutoffs were used for the categorization of low-, intermediate-, and high-score subgroups. All patients were followed up for the occurrence of RCVEs (comprising cardiovascular death, non-fatal myocardial infarction, and stroke). Cox and Poisson regression analyses were used to assess the relationship of baseline LFSs with the risk of RCVE.Results::During a mean follow-up of (54.67 ± 18.80) months, 532 (8.2%) RCVEs were recorded. Intermediate and high NFS, FIB-4, Forns, and BARD scores were independently associated with an increased risk of RCVE (hazard ratios ranging from 1.42 to 1.75 for intermediate scores and 1.35 to 2.52 for high scores). In the subgroup analyses of sex, age, BMI, DM, and hypertension status, the increased risk of RCVEs with high LFSs (NFS, FIB-4, Forns, and BARD) was maintained across the different subgroups (all P < 0.05). Conclusion::This study showed that LFSs are indeed independently associated with RCVEs, suggesting that LFSs may be used as novel tools for risk stratification in CAD patients with a prior CVE.

Objective Experimental model of experimental autoimmune Myasthenia Gravis (EAMG) were established to explore the effect of Zini-Huangqi-Gegen decoction and Peiyuan-Guben powder on EAMG rats model.Methods Experimental animals were randomly divided into the control group (n=10) and the model group (n=30).The model rats were induced by murine AChR-α97-116 peptide immunostaining for EAMG rats model.After the first immunization,the general condictions and body weight of rats were observed,and the Lennon score was used to evaluate the rats.The second immunization was performed on the 1 1th day after the first immunization.On the 15th day after the first immunization,the rats were randomly divided into the model group(n=8),Huangqi-Gegen decoction and Peiyuan-Guben powder group (abbreviated Huangpei group,n=8) and Prednisone group (n=8) according to the Lennon score.Huangpi group rats were treated with Huangqi-Gegen decoction (21.5 g/kg) combined with Peiyuan-Guben powder (0.8 g/kg),prednisone group with 0.005 4 g/kg prednisone aqueous solution,the control group and the model group oral volume of distilled water.The rats were administered with a body weight of 10 ml/kg once a day for a total of 56 days.At the 70th day after the first immunization,serum was extracted from the rats.The Anti-AChR-α97-116 IgG and its subtype in serum were detected by ELISA.The IL-4,IL-10,IL-17 in serum were detected by ELISA.Results Compared with the model group,the weight of the rats in the Huangpei group and the prednisone group significantly increased after the 28th day of the first immunization (P＜0.05).After the 36nd day of the first immunization,the Lennon score of the Huangpei group significantly decreased (P＜0.05).At the end of the administration,the amplitude of EMG amplitude attenuation (41.83％ ± 7.45％ vs.67.76％ ± 4.32％) in the Huangpei group significantly decreased (P＜0.05),and the serum IgG (1.15 ± 0.07 vs.1.24 ± 0.08),IgG1 (0.17 ± 0.01 vs.0.25 ± 0.03),IL-4 (16.54 ± 1.66 pg/ml vs.25.64 ± 1.74 pg/ml),IL-10 (113.65 ± 12.87 pg/ml vs.121.54 ± 10.44 pg/ml),IL-17 (43.58 ± 3.54 pg/ml vs.65.76 ± 3.59 pg/ml) in the rat serum significantly decreased (P＜0.05).Conclusions Huangqi-Gegen decoction and Peiyuan-Guben powder can increase the body weight of rats,decrease the concentration of AChR-Ab in serum,the concentrations of IL-4,IL-10 and IL-17 in serum,and effectively improve the symptoms of EAMG rats.