BACKGROUND:As tissue engineering brings new hope to the worldwide problem of articular cartilage repair,the construction of light-curing 3D printed hydrogel scaffolds with biomimetic composition is of great significance for cartilage tissue engineering. OBJECTIVE:To construct a biomimetic methacryloylated hyaluronic acid/acellular Wharton's jelly composite hydrogel scaffold by digital light processing 3D printing technology,and to evaluate its biocompatibility. METHODS:Wharton's jelly was isolated and extracted from human umbilical cord,then decellulated,freeze-dried,ground into powder,and dissolved in PBS to prepare 50 g/L acellular Wharton's jelly solution.Methylallylated hyaluronic acid was prepared,lyophilized and dissolved in PBS to prepare 50 g/L methylallylated hyaluronic acid solution.Acellular Wharton's jelly solution was mixed with methacrylyacylated hyaluronic acid solution at a volume ratio of 1:1,and was used as bio-ink after adding photoinitiator.Methylacrylylated hyaluronic acid hydrogel scaffolds(labeled as HAMA hydrogel scaffolds)and methylacrylylated hyaluronic acid/acellular Wharton's jelly gel scaffolds(labeled as HAMA/WJ hydrogel scaffolds)were prepared by digital light processing 3D printing technology,and the microstructure,swelling performance,biocompatibility,and cartilage differentiation performance of the scaffolds were characterized. RESULTS AND CONCLUSION:(1)Under scanning electron microscope,the two groups of scaffolds showed a three-dimensional network structure,and the fiber connection of HAMA/WJ hydrogel scaffold was more uniform.Both groups achieved swelling equilibrium within 10 hours,and the equilibrium swelling ratio of HAMA/WJ hydrogel scaffold was lower than that of HAMA hydrogel scaffold(P＜0.05).(2)CCK-8 assay showed that HAMA/WJ hydrogel scaffold could promote the proliferation of bone marrow mesenchymal stem cells compared with HAMA hydrogel scaffold.Dead/live staining showed that bone marrow mesenchymal stem cells grew well on the two groups of scaffolds,and the cells on the HAMA/WJ hydrogel scaffolds were evenly distributed and more cells were found.Phalloidine staining showed better adhesion and spread of bone marrow mesenchymal stem cells in HAMA/WJ hydrogel scaffold than in HAMA.(3)Bone marrow mesenchymal stem cells were inoculated into the two groups for chondrogenic induction culture.The results of qRT-PCR showed that the mRNA expressions of agglutinoglycan,SOX9 and type Ⅱ collagen in the HAMA/WJ hydrogel scaffold group were higher than those in the HAMA hydrogel scaffold group(P＜0.05,P＜0.01).(4)These findings indicate that the digital light processing 3D bioprinting HAMA/WJ hydrogel scaffold can promote the proliferation,adhesion,and chondrogenic differentiation of bone marrow mesenchymal stem cells.

As a potential vectored vaccine,Newcastle disease virus(NDV)has been subject to various studies for vaccine development,while relatively little research has outlined the immunomodulatory effect of the virus in antigen presentation.To elucidate the key inhibitory factor in regulating the interaction of infected dendritic cells(DCs)and T cells,DCs were pretreated with the NDV vaccine strain LaSota as an inhibitor and stimulated with lipopolysaccharide(LPS)for further detection by enzyme-linked immunosorbent assay(ELISA),flow cytometry,immunoblotting,and quantitative real-time polymerase chain reaction(qRT-PCR).The results revealed that NDV infection resulted in the inhibition of interleukin(IL)-12p40 in DCs through a p38 mitogen-activated protein kinase(MAPK)-dependent manner,thus inhibiting the synthesis of IL-12p70,leading to the reduction in T cell proliferation and the secretion of interferon-γ(IFN-γ),tumor necrosis factor-α(TNF-α),and IL-6 induced by DCs.Consequently,downregulated cytokines accelerated the infection and viral transmission from DCs to T cells.Furthermore,several other strains of NDV also exhibited inhibitory activity.The current study reveals that NDV can modulate the intensity of the innate?adaptive immune cell crosstalk critically toward viral invasion improvement,highlighting a novel mechanism of virus-induced immunosuppression and providing new perspectives on the improvement of NDV-vectored vaccine.

Articular cartilage (AC) injuries often lead to cartilage degeneration and may ultimately result in osteoarthritis (OA) due to the limited self-repair ability. To date, numerous intra-articular delivery systems carrying various therapeutic agents have been developed to improve therapeutic localization and retention, optimize controlled drug release profiles and target different pathological processes. Due to the complex and multifactorial characteristics of cartilage injury pathology and heterogeneity of the cartilage structure deposited within a dense matrix, delivery systems loaded with a single therapeutic agent are hindered from reaching multiple targets in a spatiotemporal matched manner and thus fail to mimic the natural processes of biosynthesis, compromising the goal of full cartilage regeneration. Emerging evidence highlights the importance of sequential delivery strategies targeting multiple pathological processes. In this review, we first summarize the current status and progress achieved in single-drug delivery strategies for the treatment of AC diseases. Subsequently, we focus mainly on advances in multiple drug delivery applications, including sequential release formulations targeting various pathological processes, synergistic targeting of the same pathological process, the spatial distribution in multiple tissues, and heterogeneous regeneration. We hope that this review will inspire the rational design of intra-articular drug delivery systems (DDSs) in the future.

Platycodon grandiflorum (Jacq.) A. DC. is a famous medicinal plant commonly used in East Asia. Triterpene saponins isolated from P. grandiflorum are the main biologically active compounds, among which polygalacin D (PGD) has been reported to be an anti-tumor agent. However, its anti-tumor mechanism against hepatocellular carcinoma is unknown. This study aimed to explore the inhibitory effect of PGD in hepatocellular carcinoma cells and related mechanisms of action. We found that PGD exerted significant inhibitory effect on hepatocellular carcinoma cells through apoptosis and autophagy. Analysis of the expression of apoptosis-related proteins and autophagy-related proteins revealed that this phenomenon was attributed to the mitochondrial apoptosis and mitophagy pathways. Subsequently, using specific inhibitors, we found that apoptosis and autophagy had mutually reinforcing effects. In addition, further analysis of autophagy showed that PGD induced mitophagy by increasing BCL2 interacting protein 3 like (BNIP3L) levels.In vivo experiments demonstrated that PGD significantly inhibited tumor growth and increased the levels of apoptosis and autophagy in tumors. Overall, our findings showed that PGD induced cell death of hepatocellular carcinoma cells primarily through mitochondrial apoptosis and mitophagy pathways. Therefore, PGD can be used as an apoptosis and autophagy agonist in the research and development of antitumor agents.
Humans ; Mitophagy ; Carcinoma, Hepatocellular/pathology* ; Liver Neoplasms/pathology* ; Cell Line ; Autophagy ; Apoptosis ; Membrane Proteins ; Proto-Oncogene Proteins/genetics* ; Tumor Suppressor Proteins/pharmacology*

Abstract Flexibleis an important classification of flat foot. Flatfoot occurs due to a variety of reasons and causes the medial longitudinal arch to collapse or disappear. However, many children still do not develop a normal foot arch as the grow, and a failure to intervene in a timely manner will greatly harm a child s normal mobility development. Timely detection and intervention are the key to improve the prognosis. There is a lack of uniform quantitative criteria for the diagnosis of flexible flatfoot. Currently, the commonly used diagnostic methods include physical examination, foot printing, plantar pressure test and imaging examination. This article reviews the risk factors, diagnosis, prevention and treatment of Flexible Flatfoot.

Objective To investigate the expression of R-spondin3(RSPO3)in breast cancer and its prognostic value.Methods TIMER,The Cancer Genome Atlas(TCGA),Gene ExpressionProfilling In-teractive Analysis 2(GEPIA2),Human Protein Atlas(HPA)and other databases were used to analyze the differential expression of RSPO3 in breast cancer tissues and normal tissues around cancer,and analyze the correlation between its expression level and pathological parameters of breast cancer;then the expression and structural characteristics of RSPO3 transcripts in breast cancer were investigated by GEPIA2database;Kaplan-Meier Plotter was used to plot the prognosis survival curve between RSPO3 and breast cancer patients;gene ontology(GO)and kyoto encyclopedia of genes and ge-nomes(KEGG)were used for gene function enrichment analysis and metabolic pathway analysis;TIMER database was used to analyzed the relationship between RSPO3 expression level and immune cell infiltration in tumor microenvironment.Results RSPO3 was low ex-pressed in breast cancer,and its expression level was related to the age,tumor diameter and molecular type of breast cancer patients.The low expression of RSPO3 in breast cancer was related to poor prognosis of patients;the results of GO function enrichment analysis showed that RSPO3 interacting genes were mainly enriched in biological processes involving immune cells and their receptors;the results of im-mune cell infiltration analysis showed that the expression level of RSPO3 in breast cancer tissue was positively correlated with the infiltra-tion level of CD8+T cells,CD4+T cells,CD4+T cell memory,macrophages,B cells,etc.It was negatively correlated with the infiltra-tion level of Treg;the results of KEGG metabolic pathway analysis showed that RSPO3may be involved in Wnt/β-catenin pathway.Conclusion Low expression of RSPO3 in breast cancer is related to the poor prognosis of patients,which may be involved in the progres-sion of breast cancer through inhibition of immune cell infiltration and activation of Wnt/β-catenin pathway.

Objective:To investigate the clinical application value of enhanced recovery after surgery using the LEER model in patients subjected to laparoscopic cholecystectomy in basic hospitals of Yi nationality area.Methods:Twenty-six patients who underwent laparoscopic cholecystectomy based on the concept of enhancing recovery after surgery using the LEER model in People's Hospital of Jinkouhe District of Leshan from January to October 2021 were included in the observation group. An additional 20 patients who concurrently underwent laparoscopic cholecystectomy and conventional intervention were included in the control group. Clinical efficacy, postoperative complications and postoperative pain were compared between the two groups.Results:Postoperative fasting time, length of hospital stay, and total hospital days in the observation group were 6 (6, 6) hours, 2 (2, 3) days and 4 (4, 6) days respectively, which were significantly shorter than 24 (24, 36) hours, 5 (5, 6) days, 7 (7, 9) days in the control group ( H = 351.00, 407.50, 458.00, all P < 0.05). Hospitalization cost in the observation group was 5 454.58 (5 014.11, 6 016.58) yuan, which was significantly lower than 6 611.91 (6 192.68, 7 841.73) yuan in the control group ( H = 420.00, P < 0.05). There were no significant differences in operative time and postoperative complications between the two groups (both P > 0.05). At postoperative 6 hours, Visual Analogue Scale score in the observation group was 3 (3, 4) points, and patients with mild pain accounted for 73.07% (19/26). At postoperative 24 hours, Visual Analogue Scale score in the observation group was 2 (2, 3) points, and patients with mild pain accounted for 92.31% (24/26). Overall pain was well controlled after surgery. Patient satisfaction rate in the observation was 96.15% (25/26). All patients recovered and were discharged. Conclusion:Application of enhanced recovery after surgery using the LEER model in patients subjected to laparoscopic cholecystectomy in basic hospitals of Yi nationality area can promote postoperative recovery, contribute to changing the theory of diagnosis and treatment, and improve overall medical quality. The enhanced recovery after surgery protocol using the LEER model has a good application value.

Objective:To summarize the strategy of using extracorporeal membrane oxygenation (ECMO) support during lung transplantation from 2 coronavirus disease 2019 (COVID-19) with end-stage respiratory failure.Methods:Two COVID-19 with end-stage respiratory failure patients were admitted to Nanjing Medical University Affiliated Wuxi People's Hospital in March 2020. As the homoeostasis and vital signs could not be maintained in balance by conventional treatments, lung transplantations were performed. Here, detail information about combined application of peripheral veno-venous ECMO (VV-ECMO) and central veno-arterial ECMO (CVA-ECMO) during the operation will be discussed.Results:Case 1: 59 years old, 172 cm height, 72 kg weight, who received mechanical ventilation for 22 days, tracheotomy tube for 17 days, and VV-ECMO support for 7 days. Case 2: 72 years old, 178 cm height, 71 kg weight, who received mechanical ventilation for 19 days, tracheotomy tube for 17 days, and VV-ECMO support for 18 days. As both of them have severe COVID-19-associated respiratory failure, and the recovery was determined to be unlikely, lung transplantations were performed. Severe pulmonary arterial hypertension (PAH) and cardiac insufficiency were found during the operation. Based on preoperative VV-ECMO, CVA-ECMO was added. The concomitant use of peripheral VV-ECMO and CVA-ECMO offered satisfied intraoperative oxygenation and cardiopulmonary status, the operations run smoothly, and the CVA-ECMO was successfully removed, no ECMO-related complications occurred.Conclusion:The combined use of VV-ECMO and CVA-ECMO is an optimal strategy in the end-stage ARDS patients with severe PAH and cardiac insufficiency, which can offer benefits on respiratory and cardiac functions simultaneously, and ensure surgery safety.

Objective:To investigate the main postoperative complications, causes of death and the risk factors for survival in patient with benign end-stage lung diseases within 1 year after lung transplantation.Methods:A retrospective analysis was conducted to collect the clinical data of 200 patients with benign end-stage lung disease who underwent lung transplantation admitted to Wuxi People's Hospital Affiliated to Nanjing Medical University from May 2017 to October 2018. The main postoperative complications, survival and causes of death within 1 year after operation were analyzed. The Kaplan-Meier method was used to plot survival curves, and the Log-Rank test was used to compare the influence of factors, including recipient's gender, use of marginal donor lung, primary disease, preoperative combination of moderate to severe pulmonary hypertension (PAH), intraoperative extracorporeal membrane oxygenation (ECMO) support, surgical methods, intraoperative massive blood loss, postoperative complications [infection, primary graft dysfunction (PGD), acute rejection], on 1-year survival in patients who underwent lung transplantation. The multivariate Cox proportional hazards regression model was used to evaluate the risk factors of death within 1 year after lung transplantation.Results:Two hundred patients underwent successful lung transplantation. The major postoperative complications within 1 year after transplantation included infection in 131 patients, PGD in 20 patients, acute rejection in 57 patients, anastomotic complication in 26 patients and others (new onset diabetes, osteoporosis, etc.) in 53 patients. The 3-month, 6-month, and 1-year postoperative cumulative survival rates were 81.5%, 80.0% and 77.5%, respectively. Forty-five patients died during 1 year after operation, among whom 14 died of infection, 7 died of PGD, 8 died of acute rejection, 4 died of anastomotic complication, 3 died of cardio-cerebrovascular accident, 3 died of multiple organ failure, 2 died of respiratory failure and 4 died of other causes (traffic accident, etc.). The Kaplan-Meier survival analysis showed that recipient's gender, idiopathic pulmonary fibrosis (IPF) as the primary disease, preoperative combination of moderate and severe PAH, intraoperative ECMO support, intraoperative massive blood loss, postoperative complications (infection, PGD, acute rejection) were influencing factors for postoperative 1-year survival rate. The multivariate Cox regression model showed that male was the protective factor [hazard ratio ( HR) = 0.481, 95% confidence interval (95% CI) was 0.244-0.947, P = 0.034], IPF as the primary disease ( HR = 2.667, 95% CI was 1.222-5.848, P = 0.014), intraoperative use of ECMO support ( HR = 1.538, 95% CI was 0.787-3.012, P = 0.028), massive blood loss during surgery ( HR = 2.026, 95% CI was 0.976-4.205, P = 0.045) and postoperative infection ( HR = 3.138, 95% CI was 1.294-7.608, P = 0.011), PGD ( HR = 1.604, 95% CI was 0.464-5.539, P = 0.004), and acute rejection ( HR = 1.897, 95% CI was 0.791-4.552, P = 0.015) were the independent risk factors for death within 1 year after transplantation. Conclusions:One-year survival rates after lung transplantation are affected by recipient's gender, primary disease, preoperative combination of moderate and severe PAH, intraoperative ECMO support, intraoperative massive blood loss, and postoperative complications (infection, PGD, acute rejection). The male is the protective factor, while IPF as the primary disease, intraoperative ECMO support, massive blood loss during surgery and postoperative complications (infection, PGD, acute rejection) are independent risk factors for death within 1 year after lung transplantation.

Objective:To study the level and deficiency of 25-hydroxy vitamin D [25-(OH)D] in 7215 pregnant women in Dalian district, and provide a scientific evidence for the prevention of vitamin D deficiency and reasonable supplementation.Methods:During January to December of 2018, the basic information and blood samples of 7215 pregnant were collected in Maternal and Child Health Care hospital of Dalian. The level of serum 25-hydroxy vitamin D was detected by chemiluminescence method. All data was analyzed by SPSS 17.0 software.Results:In Dalian district, the average level of serum 25-(OH)D in 7215 pregnant women was 22.74(16.02, 28.62) ng/ml, and the deficiency rate of vitamin D was 33.89%, and the severe deficiency rate was 6.68%. The average level of serum 25-(OH)D in pregnant women aged 18-24 years old, 25-34 years old and 35-45 years old were 20.22(13.61, 25.57) ng/ml, 22.75(16.12, 28.42) ng/ml and 23.60(16.76, 29.92)ng/ml, respectively. With the increasing of pregnant age, the level and the deficiency rate of serum 25-(OH)D was gradually rising with significant difference( P<0.01), and the highest deficiency rate was pregnant aged 18~24 years old with 42.40%. Conclusion:The level of vitamin D of pregnant women in Dalian district was insufficient status, especially for young pregnant women. Health education for vitamin D deficiency should be focused on young pregnant women.