Pulmonary disease is one of the major threats to human health. However, the current clinical treatment drugs for lung diseases generally have problems such as low lung delivery efficiency, fast clearance rate and obvious toxic side effects. Recently, membrane biomimetic nanocarriers have attracted more and more attention. Due to their advantages of high targeting, long cycle time, good biocompatibility and strong immune escape ability, membrane biomimetic nanocarriers have become a major research hotspot in targeted therapy of lung diseases. In this review, we discuss the main preparation methods of membrane biomimetic nanoparticles, the characteristics of membrane biomimetic nanocarriers from different cell sources and their application in the targeted therapy of lung diseases. At the same time, according to the characteristics of different membranes, the shortcomings, current technical limitations and future prospects are discussed. This review is expected to provide references for the design of membrane biomimetic nanocarriers and their potential applications in the treatment of lung diseases.

Objective To establish a renal cell co-culture system to simulate the renal barrier system,and to test its responsiveness to different glucose concentrations,and to investigate the regulatory effect of miR-29b-3p on osteopontin(OPN)/transforming growth factor β(TGF-β)pathway and the changes of this pathway under high glucose condition.Methods The three-cell co-culture system consisting of human renal podocytes,human glomerular mesangial cells and human renal tubular epithelial cells was established to test the cell viability and glucose consumption value at glucose concentrations of 5,8,12 and 16 mmol/L.The content of TGF-β and OPN in cell supernatant was measured.The recombinant plasmid and siRNA of OPN were transfected,and the expressions of TGF-β and OPN were detected by Q-PCR and Western blot.Results The mRNA expressions of OPN,TGF-β and miR-29b were significantly increased at 12 mmol/L glucose conditions.Western blot results showed that the protein expression of OPN increased in high glucose conditions,while the protein expression of TGF-β did not change significantly.After adding miR-29b-3p activator,the mRNA levels of OPN and TGF-β in the cell supernatant were significantly increased.After adding miR-29b-3p inhibitor,the mRNA levels of OPN and TGF-β in the cell supernatant were significantly decreased.Western blot results showed that compared with 5 mmol/L glucose,the protein expressions of OPN and TGF-β were increased by miR-29b-3p activator,and the protein expressions of OPN and TGF-β were decreased by miR-29b-3p inhibitor.After transfection with OPN recombinant plasmid,the content of TGF-β in the cell supernatant was significantly increased,and the mRNA expressions of OPN and TGF-β in the cells were significantly increased.After transfection with OPN siRNA,the content of TGF-β in the cell supernatant was decreased,and the expression of OPN mRNA in the cells was significantly decreased,but the expression of TGF-β mRNA was not significantly increased.Conclusion The renal cell co-culture system can mimic the complex renal environment in vivo.When induced by high glucose,cell proliferation is inhibited,glucose consumption is increased,and the content of TGF-β in the cell supernatant is increased,and miR-29b-3p has a regulatory effect on OPN/TGF-β signaling pathway in the co-culture system.

Objective:To use inductively coupled plasma mass spectrometry(ICP-MS)to screen biomarkers of element omics of thyroid cancer,and to establish a risk assessment model of element omics of thyroid cancer,so as to provide a basis for the diagnosis and treatment of thyroid cancer.Methods:A total of 200 patients with thyroid cancer who admitted to Beijing Tongren Hospital from February to November 2020 were selected as the thyroid cancer group,and 50 healthy volunteers who underwent physical examinations at hospital during the same period were selected as the healthy control group.The total amount of 28 trace elements,including iodine(I),calcium(Ca),iron(Fe),nickel(Ni),copper(Cu),zinc(Zn),selenium(Se),antimony(Sb),etc.,in their serum were determined by ICP-MS.The content of trace element,thyroid function,free triiodothyronine(FT3),free tetraiodothyronine(FT4),triiodothyronine(T3),tetraiodothyronine(T4),and thyroid volume of ultrasound examination of were analyzed,and then,a risk assessment model of elemental omics of thyroid diseases was established.Results:There were statistically significant differences in the contents of eight trace elements,including I,Ca,Fe,Ni,Cu,Zn,Se and Sb between the thyroid cancer group and the healthy control group(U=2.601,1.972,2.607,2.611,2.603,2.605,2.601,2.605,P<0.05),respectively.The I,Cr and Mn levels of female patients with thyroid cancer appeared increase,while there were significant differences in I,Mn,Fe,Ni,Cu,Zn,Se and Sb contents of male patients between the thyroid cancer group and the health control group(U=2.601,2.608,2.603,2.602,1.973,2.603,2.601,2.602,P<0.05),respectively.In thyroid cancer group,the FT3,FT4,T3,T4 correlated with I content(r=06209,0.5116,0.557,0.5923,P<0.05),respectively.There were correlations in the concentrations between Fe and Zn,between Cr and Mn,between Ca and Zn,between Se and Fe,and between Zn and Se in the thyroid cancer group(r=0.5523,0.5528,0.7158,0.5699,0.6371,0.5420,P<0.05),respectively.High concentrations of I and Mn were risk factors for thyroid cancer.The specificity and sensitivity of the risk assessment model of elemental omics of thyroid cancer were all larger than 95%.Conclusion:In patients with thyroid cancer,both of the serum Ca of female patients and serum Fe of male patients play important role besides cobalt(Co),Ni,Cu,Zn,Se and Sb play role,which can provide basis for the diagnosis and treatment of thyroid cancer.The risk assessment model based on elemental omics of thyroid cancer has favorable diagnostic performance.

Abstract: Objective　To analyze the epidemiological characteristics (season, age, gender, mixed infection and clinical manifestations, etc.) of Mycoplasma pneumoniae (MP) infection in children in Hainan Province, so as to provide epidemiological evidence-based medical basis for the prevention and control of MP infection in children in Hainan Province. Methods　The serum IgM antibodies of MP, Legionella pneumophila, Chlamydia pneumoniae, adenovirus, respiratory syncytial virus (RSV), Q fever Rickettsia, parainfluenza virus, influenza A virus and influenza B virus in children with respiratory tract infections (RTIs) who were hospitalized in pediatrics of many hospitals in Hainan Province from March 2012 to February 2020 were detected by indirect immunofluorescence method. The positive serum MP-IgM antibody was defined as MP infection. The epidemiological and clinical data of MP infected cases were analyzed retrospectively. Results　From March, 2012 to February, 2020, a total of 35 731 qualified pediatric inpatients with RTIs in many hospitals in Hainan Province were tested for serum MP-IgM with the total positive rate of 39.12% (13 978/35 731). The yearly positive rates of MP-IgM from 2012 to 2020 were 48.39%, 56.23%, 56.62%, 47.04%, 29.71%, 24.14%, 47.55%, 36.84% and 24.46% respectively. The positive rates of MP-IgM in 2013 and 2014 were significantly higher than those in other years (P<0.05). The positive rate of MP-IgM in summer in Hainan Province was the highest (41.34%) and the lowest in winter (35.77%) (P<0.05). MP infection occurred in all age groups, the positive rate of MP-IgM in children of preschool (51.80%) was significantly higher than that in other age groups (P<0.01), and the positive rate of MP IgM in children of infancy (15.36%) was lower than that in other age groups (P<0.01). The positive rate of MP-IgM in female was 44.77%, which was significantly higher than that in male (35.83%) (P<0.05). MP infection combined with positive IgM of another pathogen accounted for 32.63% (4 561 cases), positive IgM of another two pathogens accounted for 1.26% (176 cases). MP infection was mostly found in pneumonia (68.73%), and the main clinical symptoms were cough (84.72%), fever (51.01%) and wheezing (3.16%). Conclusions　MP is an important pathogen of respiratory tract infection in children in Hainan Province, and infection is more common in children in early school age and early childhood. Mp-specific tests should be performed to identify the pathogen in children suspected of MP infection. In the high incidence season, health education should be strengthened in kindergartens, schools and other places to prevent respiratory tract infection.

ObjectiveTo analyze the effects of polydatin on myeloma cell growth，apoptosis， and reactive oxygen species（ROS）/p38 mitogen-activated protein kinase（MAPK） signaling pathway. MethodHuman multiple myeloma （MM） cell line U266 cells were cultured in vitro，and the effects of polydatin at 0，20，40，80，160，200 mg·L^-1 on the growth of U266 cells were detected by cell counting kit-8（CCK-8）assay. The half-maximal inhibitory concentration（IC₅₀）was calculated. U266 cells in the logarithmic growth phase were randomly divided into a control group， low- and high-dose polydatin （80 and 160 mg·L^-1） groups， and a bortezomib （75 nmol·L^-1） group. After treatment with corresponding drugs，the cell viability of each group was determined by CCK-8 assay. The apoptosis rate of each group was measured by flow cytometry. The levels of inflammatory factors， such as tumor necrosis factor-α（TNF-α），interleukin-1β（IL-1β）， and ROS in each group were measured by enzyme-linked immunosorbent assay （ELISA）. The protein expression levels of apoptosis-related factors， including cysteine aspartate-specific protease-9（Caspase-9），B-cell lymphoma-2-associated X protein（Bax），p38 MAPK，and phosphorylated （p）-p38 MAPK in each group were detected by Western blot. ResultCompared with the results in the control group， polydatin of different concentrations could inhibit the growth of U266 cells （P<0.05），and the effect was potentiated with the increase in the concentration，with IC₅₀ of 156.54 mg·L^-1. Compared with the control group，the groups with drug treatment showed blunted cell viability （P<0.05） and increased apoptosis rate，TNF-α，IL-1β，and ROS levels， protein expression levels of Caspase-9， Bax，and p-p38 MAPK/p38 MAPK （P<0.05）. Compared with the low-dose polydatin group， the high-dose polydatin group and the bortezomib group showed improved indicators mentioned above （P<0.05）， and there was no significant difference between the high-dose polydatin group and the bortezomib group. ConclusionPolydatin can activate the ROS/p38 MAPK signaling pathway，promote the expression of inflammatory factors，inhibit the growth of U266 cells，and promote their apoptosis.

The regulation of spermatogonial proliferation and apoptosis is of great significance for maintaining spermatogenesis. The single-cell RNA sequencing (scRNA-seq) analysis of the testis was performed to identify genes upregulated in spermatogonia. Using scRNA-seq analysis, we identified the spermatogonia upregulated gene origin recognition complex subunit 6 (Orc6), which is involved in DNA replication and cell cycle regulation; its protein expression in the human and mouse testis was detected by western blot and immunofluorescence. To explore the potential function of Orc6 in spermatogonia, the C18-4 cell line was transfected with control or Orc6 siRNA. Subsequently, 5-ethynyl-2-deoxyuridine (EdU) and terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assays, flow cytometry, and western blot were used to evaluate its effects on proliferation and apoptosis. It was revealed that ORC6 could promote proliferation and inhibit apoptosis of C18-4 cells. Bulk RNA sequencing and bioinformatics analysis indicated that Orc6 was involved in the activation of wingless/integrated (Wnt)/ β-catenin signaling. Western blot revealed that the expression of β-catenin protein and its phosphorylation (Ser675) were significantly decreased when silencing the expression of ORC6. Our findings indicated that Orc6 was upregulated in spermatogonia, whereby it regulated proliferation and apoptosis by activating Wnt/β-catenin signaling.

Objective: To analyze the clinical epidemiological characteristics including composition of pathogens , clinical characteristics, and disease prognosis acute bacterial meningitis (ABM) in Chinese children. Methods: A retrospective analysis was performed on the clinical and laboratory data of 1 610 children <15 years of age with ABM in 33 tertiary hospitals in China from January 2019 to December 2020. Patients were divided into different groups according to age,<28 days group, 28 days to <3 months group, 3 months to <1 year group, 1-<5 years of age group, 5-<15 years of age group; etiology confirmed group and clinically diagnosed group according to etiology diagnosis. Non-numeric variables were analyzed with the Chi-square test or Fisher's exact test, while non-normal distrituction numeric variables were compared with nonparametric test. Results: Among 1 610 children with ABM, 955 were male and 650 were female (5 cases were not provided with gender information), and the age of onset was 1.5 (0.5, 5.5) months. There were 588 cases age from <28 days, 462 cases age from 28 days to <3 months, 302 cases age from 3 months to <1 year of age group, 156 cases in the 1-<5 years of age and 101 cases in the 5-<15 years of age. The detection rates were 38.8% (95/245) and 31.5% (70/222) of Escherichia coli and 27.8% (68/245) and 35.1% (78/222) of Streptococcus agalactiae in infants younger than 28 days of age and 28 days to 3 months of age; the detection rates of Streptococcus pneumonia, Escherichia coli, and Streptococcus agalactiae were 34.3% (61/178), 14.0% (25/178) and 13.5% (24/178) in the 3 months of age to <1 year of age group; the dominant pathogens were Streptococcus pneumoniae and the detection rate were 67.9% (74/109) and 44.4% (16/36) in the 1-<5 years of age and 5-<15 years of age . There were 9.7% (19/195) strains of Escherichia coli producing ultra-broad-spectrum β-lactamases. The positive rates of cerebrospinal fluid (CSF) culture and blood culture were 32.2% (515/1 598) and 25.0% (400/1 598), while 38.2% (126/330)and 25.3% (21/83) in CSF metagenomics next generation sequencing and Streptococcus pneumoniae antigen detection. There were 4.3% (32/790) cases of which CSF white blood cell counts were normal in etiology confirmed group. Among 1 610 children with ABM, main intracranial imaging complications were subdural effusion and (or) empyema in 349 cases (21.7%), hydrocephalus in 233 cases (14.5%), brain abscess in 178 cases (11.1%), and other cerebrovascular diseases, including encephalomalacia, cerebral infarction, and encephalatrophy, in 174 cases (10.8%). Among the 166 cases (10.3%) with unfavorable outcome, 32 cases (2.0%) died among whom 24 cases died before 1 year of age, and 37 cases (2.3%) had recurrence among whom 25 cases had recurrence within 3 weeks. The incidences of subdural effusion and (or) empyema, brain abscess and ependymitis in the etiology confirmed group were significantly higher than those in the clinically diagnosed group (26.2% (207/790) vs. 17.3% (142/820), 13.0% (103/790) vs. 9.1% (75/820), 4.6% (36/790) vs. 2.7% (22/820), χ²=18.71, 6.20, 4.07, all P<0.05), but there was no significant difference in the unfavorable outcomes, mortility, and recurrence between these 2 groups (all P>0.05). Conclusions: The onset age of ABM in children is usually within 1 year of age, especially <3 months. The common pathogens in infants <3 months of age are Escherichia coli and Streptococcus agalactiae, and the dominant pathogen in infant ≥3 months is Streptococcus pneumoniae. Subdural effusion and (or) empyema and hydrocephalus are common complications. ABM should not be excluded even if CSF white blood cell counts is within normal range. Standardized bacteriological examination should be paid more attention to increase the pathogenic detection rate. Non-culture CSF detection methods may facilitate the pathogenic diagnosis.
Adolescent ; Brain Abscess ; Child ; Child, Preschool ; Escherichia coli ; Female ; Humans ; Hydrocephalus ; Infant ; Infant, Newborn ; Male ; Meningitis, Bacterial/epidemiology* ; Retrospective Studies ; Streptococcus agalactiae ; Streptococcus pneumoniae ; Subdural Effusion ; beta-Lactamases

Consensus ; Critical Illness ; Humans ; Serum Albumin ; Serum Albumin, Human

Objective:To investigate the effect of astragaloside IV (AS-IV) on the secretion of stromal cell-derived factor-1α (SDF-1α) and CXC chemokine receptor 4 (CXCR4) by high glucose injured human umbilical vein endothelial cells (HUVECs), so as to lay a foundation for further study on AS-IV improving angiogenesis by regulating SDF-1 α/CXCR4 axis of endothelial cells.Methods:HUVECs were isolated and cultured from the umbilical vein of full-term healthy newborns and identified by von Willebrand factor (vWF) combined with 4-diamino-2-phenylindole (DAPI) nuclear staining. The obtained HUVECs was cultured in EGM-2 medium with 30 mmol/L glucose for 120 h to obtain high glucose damaged HUVECs. After intervention with different concentration gradients (25 mg/L, 50 mg/L, 100 mg/L, 200 mg/L, 400 mg/L) AS-IV for 72 hours, the contents of SDF-1α and CXCR4 were detected by enzyme linked immunosorbent assay (ELISA) method to determine the best concentration of AS-IV. The supernatant of damaged HUVECs were collected at 6, 12, 24, 48 and 72 hours after intervention with the best concentration of AS-IV, and the contents of SDF-1α and CXCR4 were detected by ELISA method to determine the best action time of AS-IV. The damaged HUVECs was randomly divided into experimental group and control group, and the blank group was set up at the same time. The experimental group was treated with the best concentration of AS-IV and the best time, the control group and the blank group were treated with the same volume of phosphate buffered saline (PBS) solution, and the contents of SDF-1α and CXCR4 in each group were detected by ELISA method.Results:The vWF factor on the cell membrane was green fluorescence, and the nucleus was blue after DAPI staining. When the fusion image showed green fluorescence, HUVECs were identified by blue fluorescence. The expression of SDF-1α in damaged HUVECs was the best when treated with AS-IV of 100 mg/L for 24 hours (1 642.87 pg/ml), and the expression of CXCR4 in damaged HUVECs was the best when treated with AS-IV of 50 mg/L for 48 hours (8.44 ng/ml). Compared with the control group, the contents of SDF-1α and CXCR4 in the experimental group were significantly increased, and the difference was statistically significant ( P<0.05). While the contents of SDF-1α and CXCR4 in the experiment group were slightly less than those in the blank group and there was no statistically significant difference ( P>0.05). Conclusions:AS-IV can promote the expression of SDF-1α and CXCR4 in HUVECs damaged by high glucose to return to normal physiological level, so as to play the role of vascular repair and neovascularization.

Objective:To investigate the impacts of astragaloside Ⅳ (AS-Ⅳ) on in- vitro proliferation and angioblastic differentiation of human umbilical cord blood-derived mesenchymal stem cells (hUCBMSCs), providing a basis for further research about the effects of AS-Ⅳ on mesenchymal stem cells (MSCs)-mediated angiogenesis. Methods:The hUCBMSCs were extracted from umbilical cord blood of normal full-term infants and subcultured. Osteoblasts, chondroblasts, and lipoblasts were induced, differentiated and identified. At the same time, the surface antigens CD44, CD73, and CD105 on hUCBMSCs were determined by flow cytometry. The successfully identified hUCBMSCs were cultured and treated with a series concentrations of AS-Ⅳ (0, 50, 100, 200, 300, and 400 mg/L). The optimum concentration of AS-Ⅳ for cell proliferation in hUCBMSCs was confirmed. In another experiment, hUCBMSCs were randomly divided into the experimental group and the control group. The cells in the experiment group were treated with the optimum concentration of AS-Ⅳ, and those in the control group were treated with equal volume of PBS. The impact of AS-Ⅳ on the proliferation of hUCBMSCs was detected using the cell counting kit (CCK-8). Besides, the impact of AS-Ⅳ on the angioblastic differentiation of hUCBMSCs was examined using the matrigel in- vitro tube formation assay. CD31 and von willebrand factor (vWF) expressions were determined using immunofluorescence after hUCBMSCs differentiated towards endothelial cells. Results:Under the light microscope, hUCBMSCs had clear edges and arranged orderly, showing a typical long fusiform structure. Flow cytometry confirmed that hUCBMSCs had surface markers of mesenchymal stem cells. The optimum concentration of AS-Ⅳ for the proliferation of MSCs was 300 mg/L. The OD values of the control and experimental groups were (0.51±0.01) and (0.98±0.05), respectively, with statistical significance ( t=15.96, P<0.05), indicating that the proliferation ability of the experimental group was enhanced. Compared with the control group, the tube density and the length of the tube network in vitro in the experimental group were higher, with statistically significant difference [(629.80±52.94)mm vs (110.36±13.19)mm, P<0.05]. Compared with the control group, the expression of CD31 and vWF increased in the experimental group after AS-Ⅳ induced hUCBMSCs differentiation ( t=13.64, 13.18, P<0.05). Conclusions:AS-Ⅳ has no toxicity to human umbilical cord blood mesenchymal stem cells, and can improve their proliferation function, and induce hUCBMSCs to differentiate into endothelial cells.