Objective To investigate the protective effect of Toll-like receptor (TLR)2/TLR9 agonists,Pam2 CSK4(Pam)and CpG ODN (CpG)on mice infected with Acinetobacter baumannii (Ab)in the lungs.Methods Female C57 mice (6～8 weeks old)were randomly divided into PBS,Pam,CpG and Pam+CpG groups.In 24 h after intranasal immunization with different doses of the corresponding agonists,the mice were given a lethal dose of Ab infection in the lungs,and the survival rates of the mice were observed.A sublethal dose lung infection model of Ab was then established,and the bacterial colonization in the blood,lungs,liver,kidneys and spleen was measured respectively in the mice after infection.HE staining was used to observe the pathological damages in the lungs and kidneys.The protective effect of the agonists in the immunized mice against Ab was examined at 1,3 and 7 d after immunization to explore the protective time window.Pam+CpG was used to stimulate A549 cells and RAW264.7 cells to investigate the killing or phagocytic effects on Ab.Results Compared to PBS,Pam+CpG treatment significantly improved the survival rate of the mice after a lethal dose of Ab lung infection (P<0.05,P<0.01 ),reduced bacterial colonization in the blood (P<0.01 ),lungs (P<0.01 ),liver (P<0.01 ),kidneys (P<0.01 )and spleen (P<0.01 )in the mice after sublethal challenge,and alleviated pathological damage caused by infection. Immunization at 1 or 3 d before infection significantly improved the survival rate (P<0.05 ),and the protective effect was the best in 3 d after immunization.Furthermore,compared to single PBS,Pam and CpG immunization,Pam+CpG significantly promoted the killing and phagocytic effects of A549 epithelial cells and RAW264.7 cells,respectively,against Ab (P<0.01 ).Conclusion Combined application of TLR2/TLR9 agonists exerts a significant protective effect on both lethal and sublethal infections of Ab,which might be by its promoting the killing or phagocytic effect of lung epithelial cells and macrophages against Ab.

Objective To evaluate the stability,safety and immune enhancement and anti-tumor effects of Wilms'tumor gene 1(WT1)peptide combined with AddaVaxTM emulsion vaccine for acute myeloid leukemia.Methods The stability of WT1 peptide in the adjuvant vaccine was evaluated using MALDI-TOF-MS time-of-flight mass spectrometry.Female C57BL/6 mice were randomly divided into PBS group,WT1 peptide group,and WT1 peptide+AddaVaxTMemulsion adjuvant vaccine group.The immunization was performed at a dose of 50 μg/mouse for antigen and 50 μg/mouse for adjuvant,with intramuscular injection on days 0,14,and 28.HE staining was used to assess the toxicity of intramuscular vaccination on mouse organ tissues.Cytokine levels were detected by ELISA,and the number of IFN-γ-secreting splenocytes was measured by ELISpot.Flow cytometry was employed to detect the maturation of bone marrow-derived dendritic cells(BMDCs)promoted by the vaccine in vitro and the promotion for lymphocyte activation,and H-2Db WT1 tetramer was utilized to detect the proportion of specific CD8+T cells.After establishing a mouse leukemia tumor model using the C1498-mWT1 stable cell line,the anti-tumor effects of the vaccine for prevention and treatment were evaluated.Results The WT1 peptide stably existed in the vaccine without causing significant organ tissue changes in mice after intramuscular injection.Compared to the mice immunized with WT1 aqueous solution,the mice after intramuscular injection of the WT1 peptide emulsion adjuvant vaccine showed stronger immune responses of Th1 cells,including IFN-γ and TNF-α,as well as Th17 cells of IL-17A(P＜0.05),and the mice had not only promoted number of IFN-γ secreting splenocytes(P＜0.01)but also enhanced maturation of BMDCs,as indicated by an increase in the proportions of CD40+/CD11c+and CD86+CD80+/CD11c+ cells(P＜0.05).Additionally,there were increases in both the proportion of CD4+/CD3+T and CD69+/CD8+T cells(P＜0.05)and the proportion of specific CD8+T cells(P＜0.05).In the anti-tumor effect study using the C1498-mWT1 mouse model,the median survival time of the WT1+AddaVaxTM group was extended by 6 d compared to the WT1 aqueous solution group.At day 50,the survival rate of mice in the WT1+AddaVaxTM group was still 28.5％,while all mice in the other groups had died(P＜0.05).Conclusion The vaccine with the WT1 peptide and AddaVaxTM emulsion adjuvant exhibits good immunological and anti-tumor effects.

Objective To investigate the antibacterial effect and its preliminary mechanism of lavender essential oil on multi-drug resistant Acinetobacter baumannii.Methods Micro-dilution method was used to determine the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC)of lavender essential oil against multi-drug resistant Acinetobacter baumannii,and bactericidal kinetic study was employed to determine the onset and maintenance time of lavender essential oil.Meanwhile,the promoting and therapeutic effects of lavender essential oil on wound healing were observed in a mouse model of infection.Subsequently,crystal violet staining was used to determine the inhibition and clearance of multi-drug resistant Acinetobacter baumannii biofilm by lavender essential oil,and laser confocal microscopy was utilized to observe the survival of bacteria in biofilms.NanoDrop instrument was utilized to quantify the leakage of bacterial DNA nucleic acid and protein after intervention with 3 and 6 mg/mL lavender essential oil,and the leakage of bacterial potassium ion was measured by potassium ion test kit.Proteomics technology combined with bio-informatics were applied to explore the action mechanism of lavender essential oil against multi-drug resistant Acinetobacter baumannii.Results The MIC and MBC of lavender essential oil were both 6 mg/mL,which could kill almost all multi-drug resistant Acinetobacter baumannii at the time point of 120 min,and showed an obvious dose-and time-dependent manner.The overall animal model evaluation showed that both 3 and 6 mg/mL lavender essential oil could promote wound healing,and the curative effect was obvious.Further studies confirmed that 3 mg/mL lavender essential oil had a certain biofilm inhibitory effect on multi-drug resistant Acinetobacter baumannii,and 6 mg/mL also had a certain biofilm clearance effect under the same conditions.Meanwhile,when incubated at 37℃ for 1 h,the dose of 3 mg/mL could increase the leakage of DNA nucleic acid and protein,and significantly promote the efflux of potassium ions.Proteomic analysis suggested that the antibacterial effect of lavender essential oil may be related to affecting the oxidorereductase activity and cell metabolic process of multi-drug resistant Acinetobacter baumannii,and interfering with the biosynthesis of cell wall/membrane/envelope and other structures.Conclusion Lavender essential oil at 3 mg/mL can play an antibacterial effect against multi-drug resistant Acinetobacter baumannii,and its mechanism may be related to the destruction of bacterial biofilm and interference with bacterial metabolism.

Objective:To investigate the correlation between the clinical features and endoscopic ultrasound-guided portal pressure gradient (EUS-PPG) in patients with cirrhosis.Methods:A total of 148 patients with cirrhosis and portal hypertension who underwent EUS-PPG measurement at the Third Xiangya Hospital of Central South University from March 15, 2022 to June 20, 2023 were selected. The clinical data of patients collected before EUS-PPG measurement were analyzed. Variations in the EUS-PPG across different clinical data subgroups were analyzed. Multivariate linear regression analysis was used to explore the independent factors influencing EUS-PPG.Results:The EUS-PPG was significantly elevated in patients exhibiting red signs (16.62±5.33 mmHg VS 13.44±5.34 mmHg, t=3.616, P<0.001), gastroesophageal varices (15.78±5.30 mmHg VS 9.70±4.77 mmHg, t=4.247, P<0.001), hepatic encephalopathy (20.83±7.52 mmHg VS 14.92±5.35 mmHg, t=2.606, P=0.010), thrombocytopenia (15.66±5.39 mmHg VS 13.29±5.83 mmHg, t=2.136, P=0.034), hypoproteinemia (16.13±5.86 mmHg VS 14.12±5.03 mmHg, t=2.230, P=0.027), and an increased international normalized ratio (16.25±6.00 mmHg VS 14.40±5.11 mmHg, t=2.022, P=0.045). Conversely, the EUS-PPG was significantly reduced in patients with a history of splenectomy and devascularization (13.17±5.88 mmHg VS 15.73±5.34 mmHg, t=-2.379, P=0.019). The EUS-PPG in patients with varying degrees of ascites (no VS slight VS moderate or severe: 13.40±5.48 mmHg VS 15.90±5.49 mmHg VS 16.69±5.17 mmHg, F=5.188, P=0.007) and different Child-Pugh classifications (A VS B VS C: 14.07±5.05 mmHg VS 15.69±5.74 mmHg VS 17.64±5.99 mmHg, F=3.066, P=0.049) increased gradually. Multivariable linear regression analysis showed that red signs ( β=2.44, t=2.732, P=0.007), gastroesophageal varices ( β=4.45, t=2.990, P=0.003), ascites ( β=1.75, t=2.368, P=0.019), and hepatic encephalopathy ( β=5.82, t=2.644, P=0.009) were independent factors for the elevated EUS-PPG. Conclusion:There is a significant correlation between EUS-PPG and the clinical features related to the severity of cirrhotic portal hypertension, which indicates the feasibility of EUS-PPG in evaluating cirrhotic portal hypertension.

Tumor has become the major reasons cause of death,and its vaccine has become the effective tracts of treatment and prevention by enhancing the immune response of patients.However,most vaccines which are recombination subunit protein antigens are poorly immunogenic and difficult to induce a robust immune response in patients with compromised immune systems,resulting in poor marketing approval.The core component of the vaccine adjuvant can greatly enhance the strength,speed and duration of the immune response,thus becoming the key to the development of an ideal tumor vaccine.Most tumor vaccines are combined with tradition adjuvant such as aluminum,MF59 and AS adjuvant,but their products and patents are monopolized by large foreign companies.We found that natural adjuvants have many unique advantages,such as good biocompatibility and biodegradability,promoting the maturation of dendritic cell and the secretion of immune cytokines,significantly enhancing the tumor vaccine immune response,etc.In this paper,the application and future development of natural polysaccharides,saponins,flavonoid and plant virus-like particles in cancer vaccines were reviewed,which may lay a solid foundation for the development of the original and innovative adjuvants with domestic independent intellectual property rights.

Objective To systematically retrieve and integrate the clinical randomized controlled trials(RCTs)and systematic reviews/Meta-analyses of traditional Chinese medicine in the treatment of diabetic peripheral neuropathy(DPN)in recent 10 years,aiming to summarize the overall evidence distribution of traditional Chinese medicine in the treatment of DPN.Methods CNKI,WANFANG,VIP,CBM,PubMed,Web of Science,Embase and the Cochrane Library were used as retrieval database.The retrieval time was from January 1,2012 to October 23,2022.RCTs and systematic reviews/Meta-analyses were included.The distribution of evidence was displayed in the form of charts.AMSTAR-1 was used for the methodological quality evaluation of systematic reviews/Meta-analyses.Results A total of 1648 RCTs and 59 systematic reviews/Meta-analyses were included.The overall number of RCTs were on the rise,but most of the scale of the RCTs were relatively small,with 68％of the samples size of a single study concentrated between 50-100;The Duration of intervention was 4-8 weeks;Multi-therapy was the most commonly used intervention,among which the most involved intervention was the combination of TCM decoction;Traditional Chinese medicine monotherapy was mainly oral traditional Chinese medicine decoction.The evaluation indexes of clinical efficacy paid much attention to the total effective rate,nerve conduction velocity,TCM diseases and syndromes;economic index,quality of life,long-term efficacy and other indicators had attracted less attention of researchers.The overall methodological quality of systematic reviews/Meta-analyses was not high,most of which show good clinical efficacy,but lack sufficient evidence support.Conclusion The research results show that the treatment of diabetes peripheral neuropathy with TCM have good characteristics and advantages,the shortcomings are mainly reflected in the low quality of the overall methodology of systematic reviews/Meta-analyses.Suggesting that more high-quality clinical RCTs with breadth and depth are still needed in the future to verify the characteristics and advantages of traditional Chinese medicine in the treatment of diabetic peripheral neuropathy and provide data information support for evidence-based medicine.

Molecular nuclear medicine is a new subject that uses nuclear medicine technology to study the changes of molecular level in organisms in order to understand their functional changes. 2021 Radiological Society of North America annual meeting has more than 40 academic reports on molecular nuclear medicine. The main content includes new tracers and new imaging methods in tumors (prostate cancer, breast cancer, rectal cancer, etc.) and other diseases (Coronavirus Disease 2019 (COVID-19), Alzheimer′s disease, Parkinson′s disease, etc.). This article reviews the relevant research progress.

Objective:To evaluate the effect of dexmedetomidine on intracranial pressure in pediatric patients undergoing laparoscopic surgery through measurement of optic nerve sheath diameter (ONSD) by ultrasound.Methods:Ninety-three pediatric patients of both sexes, aged 3-12 yr, of American Society of Anesthesiologists physical status Ⅰ or Ⅱ, with body mass index of 15.2-18.1 kg/m 2, scheduled for elective laparoscopic appendectomy, were divided into 2 groups using a random number table method: control group (group C, n=45) and dexmedetomidine group (group D, n=48). Dexmedetomidine was infused intravenously in a loading dose of 0.5 μg/kg over 10 min before induction of anesthesia followed by an infusion of 0.5 μg·kg -1·h -1 in group D, while the equal volume of normal saline was given instead in group C. ONSD was measured using ultrasound after entering the operating room (T 0), after tracheal intubation (T 1), at 5 min after pneumoperitoneum (T 2), at 5 min after trendelenburg position (T 3), at the end of anesthesia (T 4) and when orientation recovered (T 5). The development of increased intracranial pressure in pediatric patients was diagnosed according to ONSD.The time for extubation, time for recovery of orientation, the Ramsay score when orientation recovered and postoperative nausea and vomiting were recorded. Results:Compared with group C, ONSD was significantly decreased at T 2, 3, the incidence of increased intracranial pressure was decreased, and the Ramsay score when the patients recovered orientation was increased ( P<0.01), and no significant change was found in the time for extubation, time for recovery of orientation and postoperative nausea and vomiting in group D ( P>0.05). Conclusion:Dexmedetomidine can decrease the development of increased intracranial pressure and produces no effect on the postoperative recovery in pediatric patients undergoing laparoscopic surgery.

OBJECTIVE: To investigate the effects of over-expression of miR-144 on invasion of SMMC-7721 cells and Toll-like receptor (TLR)/myeloid differentiation factor 88 (MyD88) pathway in hepatocellular carcinoma cells. METHODS: The expressions of miR-144 was examined in normal human hepatocyte line HL-7702 and hepatocarcinoma cell line SMMC-7721 using realtime quantitative PCR (qRT-PCR). SMMC-7721 cells were divided into blank group, miR-144 NC group and miR-144 mimics group, and the expressions of miR-144 in each group were detected with qRT-PCR. Cell count kit-8 (CCK8) was used to assess the survival of SMMC-7721 cells, and the cell invasion was evaluated using Transwell assay. The expressions of matrix metalloproteinase-2 (MMP-2), matrix metalloproteinase-9 (MMP-9) and TLR/MyD88 pathway-related proteins in the cells were detected with Western blotting; the effect of 40 μ mol/L MyD88 inhibitor on TLR/MyD88 pathway-related proteins was examined in SMMC-7721 cells. RESULTS: Compared with normal human hepatocytes, SMMC-7721 cells expressed a significantly lower level of miR-144 ( < 0.05). CCK-8 assay showed that test showed that miR-144 over-expression significantly decreased the cell survival rate ( < 0.05), lowered the number of invasive cells, and decreased the expression of MMP-2 and MMP-9 in SMMC-7721 cells ( < 0.05). The expressions of Toll-like receptor 4 (TLR4), MyD88, phosphorylated nuclear factor-kappa B (pNF-κB) and NF-κB protein decreased significantly in miR-144 mimics group and TJ-M2010-2 group ( < 0.05) and were comparable between the two groups ( > 0.05). CONCLUSIONS Overexpression of miR-144 decreases SMMC-7721 cell survival and invasion by inhibiting TLR/MyD88 pathway.
Cell Line, Tumor ; Humans ; Liver Neoplasms ; Matrix Metalloproteinase 2 ; MicroRNAs ; Myeloid Differentiation Factor 88 ; NF-kappa B ; Signal Transduction ; Toll-Like Receptors

Objectives: To establish a geographic information application system for analyzing the spatial and temporal distribution of major infectious diseases in various regions of the world and to assess the risk of importation of those diseases, to China. Methods: We collected and integrated the following information on: 1) outbreaks and areas of epidemics of major infectious diseases in the world from 2000 to 2017, 2) cases of infectious diseases in arriving travelers through active surveillance at international entry-exit ports in mainland China from 2014 to 2016, 3) numbers of annual global international flights and travelers in the country. With the above information, a global space-time distribution database on major infectious diseases was then established, using the technology related to the system. Models regarding technologies on time-space analysis, probabilistic risk assessment and geographic information visualization, were applied to establish a geographic information system on risk assessment of infectious diseases that imported to China. Results: Through integration of information on outbreaks and epidemic areas of 60 major infectious diseases in 220 countries and regions around the world, as well as 42 kinds of infectious diseases identified among the international arrivals in mainland China, a system was then developed. Information on the distribution of major infectious diseases and their potential risks in the worldwide various regions, characteristics of spectrum and disease burden of infectious diseases imported to each province of mainland China were displayed. Thus, risks on importing infectious diseases in each province via air way were able to be evaluated and simulated by the probabilistic risk assessment model, under the information on specific kind of infectious disease, outside China. Conclusion Geographic Information System on Risk Assessment Regarding Infectious Diseases Imported to China provides basic data for epidemiological reconnaissance and assessment on risks of importing infectious diseases outside China, thus would be helpful for the improvement of strategies on surveillance, prevention and control regarding the importing infectious diseases, in China.