This study aims to investigate the protective effects and mechanisms of polysaccharide extract PCP1 from Polygonatum cyrtonema in ameliorating cerebral ischemia-reperfusion(I/R) injury in rats through modulation of the Toll-like receptor 4(TLR4)/NOD-like receptor protein 3(NLRP3) signaling pathway. In vivo, SD rats were randomly divided into the sham group, model group, PCP1 group, nimodipine(NMDP) group, and TLR4 signaling inhibitor(TAK-242) group. A middle cerebral artery occlusion/reperfusion(MCAO/R) model was established, and neurological deficit scores and infarct size were evaluated 24 hours after reperfusion. Hematoxylin-eosin(HE) and Nissl staining were used to observe pathological changes in ischemic brain tissue. Transmission electron microscopy(TEM) assessed ultrastructural damage in cortical neurons. Enzyme-linked immunosorbent assay(ELISA) was used to measure the levels of interleukin-1β(IL-1β), interleukin-6(IL-6), interleukin-18(IL-18), tumor necrosis factor-α(TNF-α), interleukin-10(IL-10), and nitric oxide(NO) in serum. Immunofluorescence was used to analyze the expression of TLR4 and NLRP3 proteins. In vitro, a BV2 microglial cell oxygen-glucose deprivation/reperfusion(OGD/R) model was established, and cells were divided into the control, OGD/R, PCP1, TAK-242, and PCP1 + TLR4 activator lipopolysaccharide(LPS) groups. The CCK-8 assay evaluated BV2 cell viability, and ELISA determined NO release. Western blot was used to analyze the expression of TLR4, NLRP3, and downstream pathway-related proteins. The results indicated that, compared with the model group, PCP1 significantly reduced neurological deficit scores, infarct size, ischemic tissue pathology, cortical cell damage, and the levels of inflammatory factors IL-1β, IL-6, IL-18, TNF-α, and NO(P<0.01). It also elevated IL-10 levels(P<0.01) and decreased the expression of TLR4 and NLRP3 proteins(P<0.05, P<0.01). Moreover, in vitro results showed that, compared with the OGD/R group, PCP1 significantly improved BV2 cell viability(P<0.05, P<0.01), reduced cell NO levels induced by OGD/R(P<0.01), and inhibited the expression of TLR4-related inflammatory pathway proteins, including TLR4, myeloid differentiation factor 88(MyD88), tumor necrosis factor receptor-associated factor 6(TRAF6), phosphorylated nuclear factor-kappaB dimer RelA(p-p65)/nuclear factor-kappaB dimer RelA(p65), NLRP3, cleaved-caspase-1, apoptosis-associated speck-like protein(ASC), GSDMD-N, IL-1β, and IL-18(P<0.05, P<0.01). The protective effects of PCP1 were reversed by LPS stimulation. In conclusion, PCP1 ameliorates cerebral I/R injury by modulating the TLR4/NLRP3 signaling pathway, exerting anti-inflammatory and anti-pyroptotic effects.
Animals ; Toll-Like Receptor 4/genetics* ; NLR Family, Pyrin Domain-Containing 3 Protein/genetics* ; Rats, Sprague-Dawley ; Rats ; Reperfusion Injury/genetics* ; Male ; Signal Transduction/drug effects* ; Polysaccharides/isolation & purification* ; Polygonatum/chemistry* ; Brain Ischemia/genetics* ; Drugs, Chinese Herbal/administration & dosage* ; Mice ; Humans

Protein tyrosine phosphatase nonreceptor type 2 (PTPN2) is a promising target for sensitizing solid tumors to immune checkpoint blockades. However, the highly polar active sites of PTPN2 hinder drug discovery efforts. Leveraging small interfering RNA (siRNA) technology, we developed a novel glutathione-responsive nano-platform HPssPT (HA/PEIss@siPtpn2) to silence PTPN2 and enhance immunotherapy efficacy in hepatocellular carcinoma (HCC). HPssPT showed potent transfection and favorable safety profiles. PTPN2 deficiency induced by HPssPT amplified the interferon γ signaling in HCC cells by increasing the phosphorylation of Janus-activated kinase 1 and signal transducer and activator of transcription 1, resulting in enhanced antigen presentation and T cell activation. The nano-platform was also able to promote the M1-like polarization of macrophages in vitro. The unique tropism of HPssPT towards tumor-associated macrophages, facilitated by hyaluronic acid coating and CD44 receptor targeting, allowed for simultaneous reprogramming of both tumor cells and tumor-associated macrophages, thereby synergistically reshaping tumor microenvironment to an immunostimulatory state. In HCC, colorectal cancer, and melanoma animal models, HPssPT monotherapy provoked robust antitumor immunity, thereby sensitizing tumors to PD-1 blockade, which provided new inspiration for siRNA-based drug discovery and tumor immunotherapy.

The use of nitrification inhibitors has been suggested as a strategy to decrease cadmium(Cd)accumulation in crops.However,the most efficient nitrification inhibitor for mitigating crop Cd accumulation remains to be elucidated,and whether and how changes in soil microbial structure are involved in this process also remains unclear.To address these questions,this study applied three commercial nitrification inhibitors,namely,dicyandiamide(DCD),3,4-dimethylpyrazole phosphate(DMPP),and nitrapyrin(NP),to pakchoi.The results showed that both DCD and DMPP(but not NP)could efficiently decrease Cd concentrations in pakchoi in urea-and ammonium-fertilized soils.In addition,among the three tested nitrification inhibitors,DMPP was the most efficient in decreasing the Cd concentration in pakchoi.The nitrification inhibitors decreased pakchoi Cd concentrations by suppressing acidification-induced Cd availability and reshaping the soil microbial structure;the most effective nitrification inhibitor was DMPP.Ammonia oxidation generates the most protons during nitrification and is inhibited by nitrification inhibitors.Changes in environmental factors and predatory bacterial abundance caused by the nitrification inhibitors changed the soil microbial structure and increased the potential participants in plant Cd accumulation.In summary,our study identified DMPP as the most efficient nitrification inhibitor for mitigating crop Cd contamination and observed that the soil microbial structural changes caused by the nitrification inhibitors contributed to decreasing Cd concentration in pakchoi.

Objective To investigate the risk factors of recurrent stroke in IS patients with CHF.Methods A total of 235 elderly IS patients with CHF admitted to Northern Jiangsu People's Hospital from January 2021 to January 2023 were enrolled in this study.According to LVEF clas-sification,they were divided into reduced ejection fraction HF group(HFrEF,LVEF＜40％,80 patients),intermediate ejection fraction HF group(HFmrEF,40％≤LVEF＜50％,57 patients),and preserved ejection fraction HF group(HFpEF,LVEF≥50％,98 patients).After all of them were followed up for 12 months,the patients with recurrent IS were assigned into the recurrent IS group(42 patients)and those without into the non-recurrent IS group(193 patients).Their gener-al data,laboratory results,clinical endpoint events and other indicators were compared in the above groups.Multivariate logistic regression analysis was applied to identify the related risk fac-tors for recurrent stroke in elderly patients with CHF and IS.Results There were significant differences in women ratio,age,history of smoking and drinking,peripheral vascular disease,RBC count,hemoglobin,hematocrit,LDL-C,prothrombin time,international standardized ratio,BNP,LVEF,LVEDD and LVESD among the three groups(P＜0.05,P＜0.01).The recurrent IS group had obviously larger proportion of diabetes mellitus,longer thrombin time,and larger LAD and RAD when compared with the non-recurrent IS group(P＜0.05,P＜0.01).Multivariate logistic regression analysis showed that diabetes(OR=0.251,95％CI:0.1 96-1.494,P=0.031)and LAD(OR=1.085,95％CI:1.008-1.169,P=0.031)were independent risk factor for recurrent stroke in elderly patients with CHF and IS.There were no significant differences in follow-up period and incidence of clinical endpoints among the three groups(P＞0.05).Conclusion Diabetes and LAD are independent risk factors for recurrent IS in elderly CHF patients with IS.

Objective To investigate the clinical features and related risk factors of chronic heart failure(CHF)patients with ischemic stroke.Methods A total of 153 elderly CHF patients with-out atrial fibrillation(AF)/atrial flutter(AFL)admitted in our hospital from January 2021 to January 2023 were enrolled,and according to diagnosis,they were divided into CHF group(84 cases)and ischemic stroke group(69 cases,CHF+ischemic stroke).General information,medical history and auxiliary examination indicators were collected.All of them were followed up for 6 months.SPSS statistical software was used to analyze the data of the 2 groups of patients,and the risk factors related to ischemic stroke were identified.Results Significant differences were observed in terms of age,hypertension,diabetes,partial thromboplastin time,left atrial diameter,left ventricular ejection fraction,right ventricular diameter,left ventri-cular end-systolic diameter,left ventricular end-diastolic diameter,CHF classification,thrombosis risk(CHA2DS2-VASc)score,HAS-BLED score,and usage of antiplatelet agents between the two groups(P＜0.05,P＜0.01).Multivariate logistic regression analysis showed that CHA2DS2-VASc score(OR=2.471,95％CI:1.356-4.502,P=0.003)and HAS-BLED score(OR=6.626,95％CI:2.049-21.432,P=0.002)were independent risk factors for ischemic stroke.ROC curve analysis indicated that the AUC value of CHA2DS2-VASc score and HAS-BLED score in predicting ischemic stroke in elderly CHF patients without AF/AFL was 0.936(95％CI:0.898-0.975,P＜0.01)and 0.922(95％CI:0.880-0.964,P＜0.01),respectively.Conclusion High CHA2DS2-VASc score and high HAS-BLED score are independent risk factors for ischemic stroke in elderly CHF patients with-out AF/AFL.

Urate oxidase (Uox) plays a pivotal role in uric acid (UA) degradation, and it has been applied in controlling serum UA level in clinical treatment of hyperuricemia (HUA). However, because Uox is a heterogenous protein to the human body, the immune rejections typically occur after intravenous administration, which greatly hampers the application of Uox-based agents. In this study, we used Lactococcus lactis NZ9000, a food-grade bacterium, as a host to express exogenous Uox genes, to generate the Uox-expressing engineered strains to treat HUA. Aspergillus flavus-derived Uox (aUox) and the "resurrected" human-derived Uox (hUox) were cloned into vector and expressed in NZ9000, to generate engineered strains, respectively. The engineered NZ9000 strains were confirmed to express Uox and showed UA-lowering activity in a time-dependent manner in vitro. Next, in an HUA mice model established by oral gavage of yeast paste, the UA levels were increased by 85.4% and 106.2% at day 7 and day 14. By contrast, in mice fed with NZ9000-aUox, the UA levels were increased by 39.5% and 48.3% while in mice fed with NZ9000-hUox were increased by 57.0% and 82.9%, suggesting a UA-lowering activity of both engineered strains. Furthermore, compared with allopurinol, the first-line agent for HUA treatment, mice fed with NZ9000-aUox exhibited comparable liver safety but better kidney safety than allopurinol, indicating that the use of engineered NZ9000 strains not only alleviated kidney injury caused by HUA, but could also avoided the risk of kidney injury elicited by using allopurinol. Collectively, our study offers an effective and safe therapeutic approach for HUA long-term treatment and controlling.
Animals ; Lactococcus lactis/metabolism* ; Urate Oxidase/genetics* ; Mice ; Uric Acid/blood* ; Hyperuricemia ; Humans ; Administration, Oral ; Aspergillus flavus/genetics* ; Male

Objective:To investigate effect and underlying lipid-lowering mechanisms of catalpol in non-alcoholic fatty liver disease(NAFLD).Methods:In vivo model of NAFLD was established with high-fat diet-fed ICR mice for 8 weeks. Low(50 mg/kg), medium(150 mg/kg), and high(300 mg/kg) doses of catalpol were administered, and the body weight, liver weight, hepatic index, and biochemical parameters of the mice were analyzed. Free fatty acid-induced LO2 in human hepatocytes to establish NAFLD cell model. Quantitative realtime PCR reaction to detect fatty acid synthesis-related gene levels. Western blotting assay was adopted to analyze proteins in the endoplasmic reticulum stress(ERS)-mediated protein kinase RNA-like endoplasmic reticulum kinase(PERK)-eukaryotic translation initiation factor 2α(eIF2α) signaling pathway. Results:Compared with model mice, body weight [(39.43±1.84)g, (34.01±1.83)g, (32.28±1.11)g vs(42.17±1.37)g, all P<0.001], liver weight [(1.03±0.06)g, (0.79±0.05)g, (0.64±0.04)g vs(1.30±0.13)g, P<0.01 or P<0.001], and liver index [(2.60±0.09)%, (2.32±0.09)%, (1.99±0.11)% vs(3.07±0.30)%, P<0.05 or P<0.001] were reduced in low, medium, and high doses of catapol model. Medium and high doses of catalpol diminished total cholesterol, triglyceride, low density lipoprotein-cholesterol, aspartate aminotransferase, and alanine aminotransferase( P<0.01 or P<0.001), increased high density lipoprotein-cholesterol( P<0.01 or P<0.001). In the cell model, elevated levels of both fatty acid synthesis genes and PERK-eIF2α pathway proteins were attenuated by catalase, and this attenuation was reversed by signaling pathway agonists. Conclusion:The Chinese herb catalpol may play a role in improving NALFD by regulating the ERS-mediated PERK-eIF2α signaling pathway.

Objective:To investigate the application and surgical effect of lobed anterolateral femoral chimeric perforator flap in repairing the complex soft tissue defects of limbs.Methods:Clinical materials of 52 cases with complex soft tissue defects of limbs were retrospectively reviewed from October 2013 to October 2020. Thirty-two males and 20 females were reviewed, and the age of them ranged from 6 to 65(mean 39) years old. The injuries include 17 cases of traffic accidents, 15 machine crush injuries, 12 engineering accidents, and 8 extensive resection of malignant tumors. The size of soft tissue defects ranged from 26.0 cm×10.0 cm to 35.0 cm×15.0 cm. Lobed anterolateral thigh chimeric perforator flaps or lobed Flow-through chimeric perforator flaps were designed according to the soft tissue defects and blood circulation of the distal extremities. Start from 8-month post-operation, upper limb function was assessed according to the Upper Limb Function Assessment Standard of Hand Surgery Branch of Chinese Medical Association and Disability of arm shoulder and hand(DASH) score scale, and Fugl-Meyer score system was used to evaluate the lower limb function. Long-term follow-up was conducted through WeChat video call and outpatient clinic evaluation.Results:All flaps survived. One patient with crush injury got delayed wound healing due to deep hemaetoma after surgery, and the rest were healed at the first stage. All the cases entered followed-up, ranged from 6 to 20(mean 8) months. The appearance of the flaps was satisfactory, and the color of the flaps has no significant difference compared with the recipient area. No recurrence of tumor was reported during the follow-up period. According to the Upper Limb Function Assessment Standard of Hand Surgery Branch of Chinese Medical Association, there were 21 excellent cases and 8 good cases, and the total excellent and good rate was 100%. The DASH score of upper limbs ranges from 11.5 to 45.1(mean 25.4±13.7), and the Fugl-Meyer score of lower limbs range from 18.6 to 31.8(mean 26.2±11.2) at the last follow-up.Conclusion:The complex soft tissue defect of limbs is a common complication caused by high-energy trauma, which is difficult to repair. The lobed anterolateral thigh chimeric perforator flap through a Decompose-Combine and arrange method for reconstruction, which is flexible in flap design. A variety of tissue flaps can be obtained by anastomosing only one group of blood vessels. It is an ideal choice for repairment of complex soft tissue defects of limbs.

Objective:To investigate the risk factors associated with anastomotic leakage after robotic surgery in mid-low rectal cancer.Methods:A retrospective case-control study method was conducted. Inclusion criteria: (1) 18 to 80 years old; (2) pathologically confirmed rectal cancer; (3) distance <10 cm from tumor to anal margin; (4) robotic anterior rectal resection. Patients with previous history of colorectal cancer surgery, distant metastases or other malignant tumors, undergoing emergency surgery, with severe abdominal adhesions or those receiving combined organ resection were excluded. Based on the above criteria, 636 patients undergoing robotic radical sphincter-preserving surgery for mid-low rectal cancer in Zhongshan Hospital from January 2015 to December 2018 were included in this study, including 398 males (62.6%) and 238 females (37.4%) with a mean age of (61.9±11.3) years. Sixty-eight cases (10.7%) received neoadjuvant chemoradiotherapy. Amony the 636 included patients, 123(19.3%) underwent natural orifice specimen extraction surgery (NOSES) and 15 (2.3%) underwent preventive stoma. According to the cirteria developed by the International Rectal Cancer Research Group in 2010, the anastomotic leakage was classified as grade A (no requirement of intervention), B (requirement of intervention), and C (requirement of operation). Logistic regression was used to analyze the relationship between anastomotic leakage and clinicopathological factors. Factors in univariate analysis with P<0.05 were included in the multivariate analysis. Results:Anastomotic leakage occurred in 38 cases (6.0%). The grading of anastomotic leakage was grade A in 13 cases (2.0%), grade B in 19 cases (3.0%), and grade C in 6 cases (0.9%). The 3-year disease-free survival rate of patients with anastomotic leakage and without anastomotic leakage was 83.5% and 83.6% respectively ( P=0.862); the 3-year overall survival rate of the two group was 85.1% and 87.5% respectively ( P=0.296). The results of univariate logistic regression analysis showed that male ( P=0.011), longer operation time ( P=0.042), distance ≤5 cm from tumor to anal margin ( P=0.012), more intraoperative blood loss ( P=0.048) were associated with anastomotic leakage (all P<0.05). NOSES was not associated with anastomotic leakage ( P=0.704). Multivariate analysis confirmed that male (OR=3.03, 95%CI: 1.37 to 7.14, P=0.010), operation time ≥180 minutes (OR=2.04, 95%CI: 1.03 to 3.99, P=0.040), distance ≤5 cm from tumor to anal margin (OR=2.56, 95%CI:1.28 to 5.26, P=0.008) were independent risk factors for anastomotic leakage. Conclusion:Male, short distance from tumor to anal margin, and long operation time are independent risk factors for anastomotic leakage in patients undergoing robotic mid-low rectal cancer radical surgeries. These patients need to be cautiously treated during surgery.

Objective:To investigate the risk factors associated with anastomotic leakage after robotic surgery in mid-low rectal cancer.Methods:A retrospective case-control study method was conducted. Inclusion criteria: (1) 18 to 80 years old; (2) pathologically confirmed rectal cancer; (3) distance <10 cm from tumor to anal margin; (4) robotic anterior rectal resection. Patients with previous history of colorectal cancer surgery, distant metastases or other malignant tumors, undergoing emergency surgery, with severe abdominal adhesions or those receiving combined organ resection were excluded. Based on the above criteria, 636 patients undergoing robotic radical sphincter-preserving surgery for mid-low rectal cancer in Zhongshan Hospital from January 2015 to December 2018 were included in this study, including 398 males (62.6%) and 238 females (37.4%) with a mean age of (61.9±11.3) years. Sixty-eight cases (10.7%) received neoadjuvant chemoradiotherapy. Amony the 636 included patients, 123(19.3%) underwent natural orifice specimen extraction surgery (NOSES) and 15 (2.3%) underwent preventive stoma. According to the cirteria developed by the International Rectal Cancer Research Group in 2010, the anastomotic leakage was classified as grade A (no requirement of intervention), B (requirement of intervention), and C (requirement of operation). Logistic regression was used to analyze the relationship between anastomotic leakage and clinicopathological factors. Factors in univariate analysis with P<0.05 were included in the multivariate analysis. Results:Anastomotic leakage occurred in 38 cases (6.0%). The grading of anastomotic leakage was grade A in 13 cases (2.0%), grade B in 19 cases (3.0%), and grade C in 6 cases (0.9%). The 3-year disease-free survival rate of patients with anastomotic leakage and without anastomotic leakage was 83.5% and 83.6% respectively ( P=0.862); the 3-year overall survival rate of the two group was 85.1% and 87.5% respectively ( P=0.296). The results of univariate logistic regression analysis showed that male ( P=0.011), longer operation time ( P=0.042), distance ≤5 cm from tumor to anal margin ( P=0.012), more intraoperative blood loss ( P=0.048) were associated with anastomotic leakage (all P<0.05). NOSES was not associated with anastomotic leakage ( P=0.704). Multivariate analysis confirmed that male (OR=3.03, 95%CI: 1.37 to 7.14, P=0.010), operation time ≥180 minutes (OR=2.04, 95%CI: 1.03 to 3.99, P=0.040), distance ≤5 cm from tumor to anal margin (OR=2.56, 95%CI:1.28 to 5.26, P=0.008) were independent risk factors for anastomotic leakage. Conclusion:Male, short distance from tumor to anal margin, and long operation time are independent risk factors for anastomotic leakage in patients undergoing robotic mid-low rectal cancer radical surgeries. These patients need to be cautiously treated during surgery.