ObjectiveTo investigate the effect of Tougu Xiaotong capsules （TGXTC） on the regulation of chondrocyte PANoptosis， delay of chondrocyte degeneration， and improvement of the symptoms in knee osteoarthritis （KOA）. MethodsIn vivo experiments： 50 male C57BL/6 mice were randomly assigned into five groups （n=10 per group）： sham operation group， model group， low-dose TGXTC group （7.2 g·kg^-1）， high-dose TGXTC group （14.4 g·kg^-1）， and diclofenac sodium group （0.05 g·kg^-1）. Except for the sham group， KOA models were established in all other groups using the modified Hulth method. Following successful model induction， the TGXTC groups received daily oral gavage of 7.2 or 14.4 g·kg^-1 for 6 weeks， while the diclofenac sodium group received 0.05 g·kg^-1 solution daily over the same duration. Model evaluation was performed using Lequesne MG score； micro-computed tomography （micro-CT） was used to scan the knee， hematoxylin-eosin （HE） staining and safranin O-fast green staining were used to observe the morphology of cartilage， transmission electron microscopy （TEM） was used to determine ultrastructural changes of PANoptosis. Multiple immunofluorescence （IF） co-localization assays was performed to detect the co-localization of cleaved Caspase-3， receptor-interacting protein 3 （RlPK3）， and the N-terminal domain of gasdermin D （GSDMD-N） in cartilage tissue， while western blot was employed to detect the expression levels of cleaved Caspase-3， RIPK3， and GSDMD-N. In vitro experiments： The knee cartilages of 4-week-old SD rats were isolated， and a chondrocyte in vitro culture system was established through mechanical digestion with 0.2% type Ⅱ collagenase. Second-generation chondrocytes were divided into three groups： the control group， the model group （pretreated with 10 mg·L^-1 lipopolysaccharide （LPS） for 24 h followed by treatment with 1 μmol·L^-1 nigericin for 4 h）， and the TGXTC treatment group （pretreated with 10 mg·L^-1 LPS for 24 h， followed by exposure to 1 μmol·L^-1 nigericin for 4 h and subsequently treated with 100 mg·L^-1 TGXTC for an additional 24 h）. The levels of reactive oxygen species （ROS）， apoptosis， necroptosis， and pyroptosis of chondrocytes were evaluated via fluorescence microscopy following staining with ROS detection， AO/EB and YO-PRO-1/PI staining kits. Transmission electron microscopy was utilized to investigate the ultrastructural changes associated with PANoptosis in cartilage tissue of KOA mice. Inflammatory cytokine levels （IL-1β and IL-18） were measured using ELISA. Western blot was conducted to assess protein expressions related to PANoptosis， including cleaved Caspase-3， cleaved Caspase-8， RIPK3， ZBP1， GSDMD-N， and NLRP3. ResultsCompared with the sham group， the Lequesne MG scores were significantly up-regulated（P<0.01） in the model group， and the pathological changes of cartilage were significantly， with joint spaces narrower， osteophyte formation increased， secere abrasion of cartilage surface. Ultrastructural analysis revealed pronounced chondrocyte apoptosis， necroptosis， and pyroptosis， along with markedly elevated expression of cleaved Caspase-3， RlPK3， and GSDMD-N in cartilage tissue （P<0.01）. In addition， The mean fluorescence intensities of ROS， orange-red fluorescence in AO/EB staining， green fluorescence and red fluorescence in YO-PRO-1/PI staining were increased of chondrocyte in the model group （P<0.01） . The levels of inflammatory factors IL-1β and IL-18 in the supernatant were increased （P<0.01）. The expression of PANoptosis related proteins （cleaved Caspase-3， cleaved Caspase-8， RIPK3， ZBP1， GSDMD-N， and NLRP3） were also significantly upregulated（P<0.05）. Compared to the model group， the TGXTC group demonstrated a significant improvement in various parameters of mice. These included a reduction in the Lequesne MG score， an increase in joint space， a decrease in osteophyte formation， diminished cartilage damage， reduced release of ROS， and alleviation of apoptotic， necroptotic， and pyroptotic processes in chondrocytes. Additionally， mitochondrial swelling and endoplasmic reticulum dilation were also mitigated. The levels of ROS as well as IL-1β and IL-18 were significantly decreased （P<0.05）. Furthermore， the expression levels of proteins associated with PANoptosis in cartilage tissue showed marked reductions （P<0.05）. Similar results were observed in chondrocytes： cleaved Caspase-3， cleaved Caspase-8， RIPK3， ZBP1， GSDMD-N， and NLRP3 exhibited significant decreases as well （P<0.05）. ConclusionTGXTC may mitigate chondrocytes degeneration and alleviate KOA symptoms by reducing oxidative stress and suppressing the activation of PANoptosis pathways.

ObjectiveTo investigate the effect of Tougu Xiaotong capsules （TGXTC） on the regulation of chondrocyte PANoptosis， delay of chondrocyte degeneration， and improvement of the symptoms in knee osteoarthritis （KOA）. MethodsIn vivo experiments： 50 male C57BL/6 mice were randomly assigned into five groups （n=10 per group）： sham operation group， model group， low-dose TGXTC group （7.2 g·kg^-1）， high-dose TGXTC group （14.4 g·kg^-1）， and diclofenac sodium group （0.05 g·kg^-1）. Except for the sham group， KOA models were established in all other groups using the modified Hulth method. Following successful model induction， the TGXTC groups received daily oral gavage of 7.2 or 14.4 g·kg^-1 for 6 weeks， while the diclofenac sodium group received 0.05 g·kg^-1 solution daily over the same duration. Model evaluation was performed using Lequesne MG score； micro-computed tomography （micro-CT） was used to scan the knee， hematoxylin-eosin （HE） staining and safranin O-fast green staining were used to observe the morphology of cartilage， transmission electron microscopy （TEM） was used to determine ultrastructural changes of PANoptosis. Multiple immunofluorescence （IF） co-localization assays was performed to detect the co-localization of cleaved Caspase-3， receptor-interacting protein 3 （RlPK3）， and the N-terminal domain of gasdermin D （GSDMD-N） in cartilage tissue， while western blot was employed to detect the expression levels of cleaved Caspase-3， RIPK3， and GSDMD-N. In vitro experiments： The knee cartilages of 4-week-old SD rats were isolated， and a chondrocyte in vitro culture system was established through mechanical digestion with 0.2% type Ⅱ collagenase. Second-generation chondrocytes were divided into three groups： the control group， the model group （pretreated with 10 mg·L^-1 lipopolysaccharide （LPS） for 24 h followed by treatment with 1 μmol·L^-1 nigericin for 4 h）， and the TGXTC treatment group （pretreated with 10 mg·L^-1 LPS for 24 h， followed by exposure to 1 μmol·L^-1 nigericin for 4 h and subsequently treated with 100 mg·L^-1 TGXTC for an additional 24 h）. The levels of reactive oxygen species （ROS）， apoptosis， necroptosis， and pyroptosis of chondrocytes were evaluated via fluorescence microscopy following staining with ROS detection， AO/EB and YO-PRO-1/PI staining kits. Transmission electron microscopy was utilized to investigate the ultrastructural changes associated with PANoptosis in cartilage tissue of KOA mice. Inflammatory cytokine levels （IL-1β and IL-18） were measured using ELISA. Western blot was conducted to assess protein expressions related to PANoptosis， including cleaved Caspase-3， cleaved Caspase-8， RIPK3， ZBP1， GSDMD-N， and NLRP3. ResultsCompared with the sham group， the Lequesne MG scores were significantly up-regulated（P<0.01） in the model group， and the pathological changes of cartilage were significantly， with joint spaces narrower， osteophyte formation increased， secere abrasion of cartilage surface. Ultrastructural analysis revealed pronounced chondrocyte apoptosis， necroptosis， and pyroptosis， along with markedly elevated expression of cleaved Caspase-3， RlPK3， and GSDMD-N in cartilage tissue （P<0.01）. In addition， The mean fluorescence intensities of ROS， orange-red fluorescence in AO/EB staining， green fluorescence and red fluorescence in YO-PRO-1/PI staining were increased of chondrocyte in the model group （P<0.01） . The levels of inflammatory factors IL-1β and IL-18 in the supernatant were increased （P<0.01）. The expression of PANoptosis related proteins （cleaved Caspase-3， cleaved Caspase-8， RIPK3， ZBP1， GSDMD-N， and NLRP3） were also significantly upregulated（P<0.05）. Compared to the model group， the TGXTC group demonstrated a significant improvement in various parameters of mice. These included a reduction in the Lequesne MG score， an increase in joint space， a decrease in osteophyte formation， diminished cartilage damage， reduced release of ROS， and alleviation of apoptotic， necroptotic， and pyroptotic processes in chondrocytes. Additionally， mitochondrial swelling and endoplasmic reticulum dilation were also mitigated. The levels of ROS as well as IL-1β and IL-18 were significantly decreased （P<0.05）. Furthermore， the expression levels of proteins associated with PANoptosis in cartilage tissue showed marked reductions （P<0.05）. Similar results were observed in chondrocytes： cleaved Caspase-3， cleaved Caspase-8， RIPK3， ZBP1， GSDMD-N， and NLRP3 exhibited significant decreases as well （P<0.05）. ConclusionTGXTC may mitigate chondrocytes degeneration and alleviate KOA symptoms by reducing oxidative stress and suppressing the activation of PANoptosis pathways.

Objective:To analyze the expression and prognosis of B-cell lymphoma 7 protein family member A (BCL7A) in hepatocellular carcinoma, as well as the effect and mechanism of BCL7A expression on the invasion and migration of hepatocellular carcinoma cells.Methods:The cancer tissues and adjacent tissues of 40 patients with hepatocellular carcinoma who underwent radical hepatobiliary resection in the Department of Hepatobiliary Surgery, Affiliated Hospital of Nantong University from November 2017 to March 2018 were prospectively collected for protein extraction, including 29 males and 11 females, aged (58.5±10.4) years. The information of 374 cases of hepatocellular carcinoma and 50 cases of adjacent tissues were downloaded from The Cancer Genome Atlas (TCGA) database, and the hepatocellular carcinoma cell lines Hep3B and SMMC-7721 were transfected with overexpressing BCL7A plasmid and empty vector plasmid (negative control), respectively. Western blotting and immunohistochemistry were used to detect the expression of BCL7A, and Western blotting was also used to detect the expression of proteins related to epithelial-mesenchymal transition (N-cadherin, E-cadherin, snail). Transwell and cell scratch assays were used to detect cell invasion and migration.Results:Compared with adjacent tissues, the mRNA expression of BCL7A in 50 patients with hepatocellular carcinoma in TCGA was significantly increased ( t=13.38, P<0.001). According to the median mRNA expression level of BCL7A, 374 patients were divided into BCL7A high expression group ( n=187) and low expression group ( n=187), and the cumulative survival rate of BCL7A high expression patients was lower than that of low expression group, and the difference was statistically significant ( χ2=6.95, P=0.009). Western blot was used to detect the relative expression of BCL7A protein in cancer tissues, and found it was higher compared to adjacent tissues. Compared with the negative control group, the number of cells invaded by the BCL7A overexpression group of hepatoma cells Hep3B and SMMC-7721 was more than the negative control group respectively, (153.7±1.3) vs (63.7±4.7) and (307.7±25.14) vs (72.3±12.5), and the differences were statistically significant ( t=7.97, 8.38, both P=0.001) .The results of the cell scratch assay were consistent with the results of the Transwell invasion assay. The expressions of N-cadherin and snail in the BCL7A overexpression group were higher than those in the negative control group, and the E-cadherin was lower, and the difference was statistically significant (all P<0.05). Conclusions:The expression of BCL7A in cancer tissues of patients with hepatocellular carcinoma is elevated and is associated with poor prognosis. BCL7A may promote hepatocellular carcinoma cell metastasis and invasion by promoting epithelial-mesenchymal transition.

Objective:To investigate the effect of early protein supplementation on the clinical outcomes of elderly ICU patients with critically ill.Methods:The study was a post-hoc analysis of a multicenter, cluster randomized controlled trial (NEED trial), which aimed to evaluate the impact of feeding protocol on nutritional implementation and outcomes in ICU patients. It was planned to include elderly patients aged ≥70 years from the NEED trial, and patients who had not started nutritional therapy by the Day 3 after enrolment, stayed in the ICU less than 7 days, missing the primary outcome were excluded. The primary outcome of this study was 28-day mortality of enrolment. Patients were categorized into Q1 (<0.6 g/kg/d), Q2 (0.6-0.83 g/kg/d), and Q3 (≥0.83 g/kg/d) groups according to the tertiles of protein supply. The log-rank test was used to compare the Kaplan-Meier survival curves for 28-day mortality. The associations between different protein groups and 28-day mortality were tested by Cox proportional hazards regression models. Subgroup analysis was conducted in patients with high (mNUTRIC score≥5) nutritional risk or patients with baseline acute kidney injury.Results:A total of 789 elderly (≥70 years) patients was included in the study, with a mean protein amount of 0.69 (0.53, 0.91) g/(kg·d) during days 3-7 after ICU admission, and mean protein amounts in the Q1 low-protein group, the Q2 medium-protein group, and the Q3 high-protein group were 0.46 (0.36, 0.53), 0.69 (0.63, 0.76), and 1.03 (0.91, 1.23) g/(kg·d), respectively. The results showed that the medium protein group associated with lower 28-day mortality compared to the high protein group, and the association between the medium protein group and lower 28-day mortality still held after controlling for possible confounders by Cox multivariate regression analysis. In the high-nutritional risk subgroup (mNUTRIC≥5), a significant association was also found between the medium protein group and lower 28-day mortality.Conclusions:Early high protein supply are not beneficial for elderly ICU patients by this large sample size post-hoc analysis, and medium protein supply associate with lower 28-day mortality compared with the high protein group. This study may provide a theoretical basis for the optimal dose of early protein supply in elderly ICU patients, as well as a reference for clinical implementation.

【Objective】 To establish and verify a nomogram model of overall survival (OS) of prostate cancer patients based on the SEER data. 【Methods】 A total of 12 642 patients diagnosed with prostate cancer during 2010 and 2015 were extracted from the SEER database. Patients were randomly divided into the model group (n=8 850) and validation group (n=3 792). The independent risk factors for OS were analyzed with univariate Cox proportional risk regression, lasso regression and multivariate Cox proportional risk regression. A nomogram was constructed to predict the 1-year, 3-year and 5-year OS. The prediction potential of the model was evaluated with the consistency index (C-index), calibration curve and receiver operating characteristic (ROC) curve. 【Results】 Multivariate Cox regression analysis showed that age, T stage, N stage, M stage, bone metastasis, liver metastasis and regional lymphadenectomy were independent risk factors for OS (P<0.05). The seven factors were used to construct an OS nomogram model. The C-index of the modeling set was 0.750, and the area under the ROC curve (AUC) at 1, 3 and 5 years were 0.77, 0.77 and 0.76, respectively;the C-index of the validation set was 0.765, and the AUC at 1, 3 and 5 years were 0.83, 0.79 and 0.76, respectively. The calibration curves of the modelling set and validation set showed a good agreement with the actual survival prediction rate. Risk stratification of patients based on the nomogram model showed that the OS of patients in the high-risk group was significantly lower than that in the low-risk group (P<0.001). 【Conclusion】 The nomogram can be used to predict the prognosis of prostate cancer patients, and is important for individualized treatment plans.

Induction of immunogenic cell death promotes antitumor immunity against cancer. However, majority of clinically-approved drugs are unable to elicit sufficient ICD. Here, our study revealed that mitochondria-targeted delivery of doxorubicin (DOX) massively amplified ICD via substantial generation of reactive oxygen species (ROS) after mitochondrial damage. The underlying mechanism behind increased ICD was further demonstrated to be ascribed to two pathways: (1) ROS elevated endoplasmic reticulum (ER) stress, leading to surface exposure of calreticulin; (2) ROS promoted release of various mitochondria-associated damage molecules including mitochondrial transcription factor A. Nevertheless, adaptive upregulation of PD-L1 was found after such ICD-inducing treatment. To overcome such immunosuppressive feedback, we developed a tumor stimuli-responsive nano vehicle to simultaneously exert mitochondrial targeted ICD induction and PD-L1 blockade. The nano vehicle was self-assembled from ICD-inducing copolymer and PD-L1 blocking copolymer, and possessed long-circulating property which contributed to better tumor accumulation and mitochondrial targeting. As a result, the nano vehicle remarkably activated antitumor immune responses and exhibited robust antitumor efficacy in both immunogenic and non-immunogenic tumor mouse models.

Tooth extraction in patients receiving anticoagulation/antiplatelet therapy is often considered contraindicated by many oral and maxillofacial surgeons because of a higher risk of postoperative bleeding. Multiple factors contribute to postoperative bleeding, but there is no consensus. Based on recent literature, this article reviews factors related to bleeding after tooth extraction in patients receiving anticoagulation/antiplatelet therapy. The literature review indicates that patients taking antiplatelet drugs have a lower postoperative bleeding risk than patients taking anticoagulant drugs. Prescription of anticoagulants together with non-steroidal anti-inflammatory drugs, selective serotonin inhibitors or serotonin-norepinephrine inhibitors increases the risk of bleeding, so does preoperative antibiotic use increase. In addition, systemic diseases such as diabetes, history of infection at the extraction site, and greater surgical trauma are associated with a higher risk of postoperative bleeding. At present, it is generally believed that it is safe and feasible to use different hemostatic measures after tooth extraction and to rationally apply different hemostatic measures after surgery. More prospective controlled trials are needed in the future to establish an assessment system for patients undergoing anticoagulation/antiplatelet therapy under different conditions during tooth extraction.

Objective:To explore the predictive factors of renal replacement therapy (RRT) in extracorporeal membrane oxygenation (ECMO) patients.Methods:The clinical data of 68 ECMO patients treated at Emergence Department of Jiangsu Provincial Hospital from January 2015 to December 2018 were retrospectively analyzed. Vasoactive-inotropic score (VIS) was used to assess the usage of vasoactive-inotropic drugs on day 1, 2 and 3 of ECMO (24 h VIS, 48 h VIS, and 72 h VIS). According to received RRT or not, patients were divided into the RRT group and non-RRT group. Age, gender, weight, VIS, presence of cardiac arrest before ECMO, ECMO mode, and ECMO treatment time were compared. Logistic regression analysis was used to identify predictive factors for RRT in ECMO patients.Results:Of the enrolled patients, 73.5% of ECMO patients received RRT. The mean age, 24 h VIS, ECMO failure and mortality of the RRT group were significantly higher than those of the non-RRT group ( P <0.05). The use of RRT was 87.8% in elderly ECMO patients (> cutoff age of 38.5 years). According to the cutoff value of 24 h VIS (33.75), ECMO patients were divided into the high VIS group and low VIS group. The rates of RRT and mortality were both exceeded 90% in the high VIS group, which was significantly higher than that of the low VIS group ( P <0.05). Logistic regression analysis showed that age ( OR=1.223) and 24 h VIS ( OR=1.033) were predictive factors of RRT in ECMO patients ( P <0.05). Conclusions:Age and 24 h VIS show the predictive value for RRT in ECMO patients.

Objective: To explore the function of cluster needling at scalp points therapy on regulating differential protein's expression at different time points in middle cerebral artery occlusion (MCAO) model rats. Methods: Fifty-four rats were divided into three groups randomly and 18 rats in each group. The groups respectively were the model group (group M, n = 18), cluster needling at scalp points group (group C, n = 18), false operation group (group F, n = 18). Each group was then assigned in three subgroups, including 24-h, 7-day, and 14-day subgroups. Six rats in each subgroup. Acupuncture at Baihui (GV20) and 2 points beside Baihui, which was 3-4 mm away from the midline. Longa score was used to eval-uated neurological effects. Proteomics methods were used to identify differentially expression proteins with a standard of fold change greater than 1.5 and P<.05 at different times. Results: 1. Nerve function scoring: The nerve function scores at 7 and 14 days decreased in group C, which showed better neural function than group M (P<.05). 2. Fold change in proteins:Group M showed 932 differentially expressed proteins compared with group F, and among them, 414 proteins showed significant changes in expression after acupuncture. The expression levels of Cdc42 and GFAP were increased, and Mag, Shank2, and MBP levels were decreased. In the Gene Ontology analysis, the cellular component consisted of the terms cytoplasm, cytoskeleton, lysosome, and plasma membrane. The main related biological processes were cell-cell signaling, protein transport, aging, and cell adhesion. Many synaptic and metabolic pathways were found by KEGG analysis. Conclusion: Cluster needling at scalp acupoints can improve the nerve function score and improve dyskinesia in MCAO model rats. Cluster needling at scalp acupoints can regulate the expression of 414 proteins, including Cdc42, GFAP, Mag, Shank2, and MBP, which are related to cerebral ischemia. The differential proteins are major concentration in cytoplasm, cytoskeleton, lysosomes, and plasma mem-brane, participate in cell-cell signaling, protein transport, aging, and cell adhesion, and act through multiple synaptic and metabolic pathways to exert their biological functions.

Objective To explore the relationship among depression,anxiety and social support in elderly patients in community outpatient clinic. Methods A total of 551 elderly outpatients from two com-munity health service centers of Hongkou District in Shanghai were evaluated with patient health question-naire-9 (PHQ-9),generalized anxiety disorder-7 (GAD-7),perceived social support scale( PSSS) for de-pression,anxiety,physical health and social support. Results The prevalence rates of depression and anxiety were 26. 1% and 17. 2%,respectively. The scores of PHQ-9 and GAD-7 were 2. 0(4. 0) and 1. 0(2. 0). There were statistically significant differences in the scores of family support,friend support,other support and social support among the elderly patients with different degrees of depression or anxiety (P<0. 01). Fam-ily support(B=-0. 196) and friend support(B=-0. 171) were protective factors of depression in elderly pa-tients in community outpatient clinic. Age,family support and friend support were protective factors of anxiety in elderly patients,while gender and fluctuation of physical diseases were protective factors of anxiety(P<0. 05). Con-clusions The depression and anxiety is intimately related to social support in elderly outpatients. Appropriate measures should be taken to optimize social support,mitigate bad mood negative improve their quality of life.