Objective To study the relationship between the selection of tube tension in digitalchest radiograph and image quality.Methods When tube current was fixed at 4 mAs, the choice of X-ray tube voltage changed from 60 to 120 kV.CDRAD2.0 contrast details phantom and normal human chest were exposed by X-ray system with 7 kinds of tube voltage (the difference between tube voltage was 10 kV) ,and the X-ray incidental dose of phantom surface was measured.Five radiologists with 3 years working experience evaluated the image quality on monitor and calculated the image quality factor (IQF) and image reading score.Statistics analysis was then performed by using ANOVA test and t test.According to the results, the optimum tube voltage range was determined.Results (1) The incidental dose of phantom surface increased with the higher tube voltage.(2)When the tube voltage was changed from 60 kV to 120 kV, the IQF value observed in CDRAD2.0 phantom image on monitor was 75.0±10.4,57.1±6.4,52.7±2.5,47.9±4.5, 46.0±3.8,46.0±2.8,45.2±3.5 ,there was significant statistical differences between groups(F=19.10, P<0.01).(3)The integrated score of the chest image quality in the tube voltage 90 kV and 120 kV were 12.4±0.9 and 13.0±0.7, respectively, and there was no statistical difference between two groups(t= 1.500,P>0.05).Conclusions (1)With the increase of tube tension,the display capacity of display device gradually strengthened.When the tube tension exceeded 90 kV, the increase of image quality on monitor was not evident.(2) With proper radiation dose, the value of tube tension in digital X-ray chest photograph was about 90 kV.

Objective To investigate the clinical characteristics of the human Adenovirus (HAdv) infections in allogeneic hematopoietic stem cell transplantation ( allo-HSCT) patients and explore the clinical significance of HAdv monitoring .Methods A total of 845 cases underwent allo-HSCT were included retrospectively in Perking University People′s Hospital from October 2012 to August 2014.Peripheral blood HAdv load were monitored twice weekly within 100 days after allo-HSCT, or whenever necessary quantitatively by real-time PCR. Meanwhile, other clinical samples such as stool , urine, and bronchoalveolar lavage fluid ( BLAF ) were also detected qualitatively whenever necessary .The follow-up period was at least six months after allo-HSCT.All clinical data were collected and analyzed .Results The total positive rate of HAdv was 3.4% ( 29/845 ) .The incidence of HAdv infection was higher in children [3.8%(6/155), <18y] than that of adults [3.3%(23/690),≥18y].HAdv infection diagnosed within 100 days after allo-HSCT accounted for 72.4%(21/29) of the total number of positive cases .There were 19 cases detected positive in peripheral blood , 16 cases in stool , 9 cases in urine , and 1 cases in BLAF , respectively.One patient was positive in peripheral blood , stool and urine.The overall median time of HAdv was 69 (13-189) d.The median time was 56 (53 -144) d in stool ,which was earlier than that of in peripheral blood , urine and stool.Among 29 cases of HAdv positive patients , 17 patients were coinfected with Cytomegalovirus(CMV) and 11 casess with Epstein-Barr virus(EBV).Twenty-five cases of HAdv were diagnosed with acute graft-versus-host disease(aGVHD) before HAdv infection, and 4 cases were diagnosed with chronic graft-versus-host disease ( cGVHD ) . The most common clinical manifestation was HAdv enteritis (14 cases), followed by hemorrhagic cystitis (7 cases).Two cases complicated with multiple organ injury ( >2 ) clinically, 1 cases with pneumonia.There were 8 cases of death at the end of follow-up.Conclusions HAdv is an important pathogen causing infection in patients after allo-HSCT. The infenction is characterized with multiple organ involvement .CMV and EBV coinfection is common .HAdv monitoring was of great significance in allo-HSCT patients.

Objective　To explore the effect of Mp1p on host macrophages through transcriptomics combined with metabolomics. Methods Firstly, a THP-1 macrophage strain (THP-1-Mp1p+) stably expressing Mp1p was constructed using lentivirus. Secondly, using high-throughput RNA sequencing (RNA Seq) technology, the expression level of intracellular mRNA was detected in transcriptomics analysis to determine differentially expressed genes; In metabolomics analysis, metabolite identification was performed through database comparison, and pathway analysis was performed on differential metabolites to reveal potential mechanisms of action. Finally, the results of metabolomics and transcriptomics were combined for analysis, and differential metabolites and genes were analyzed to further elucidate the mechanism of action of Mp1p on macrophages. Results Transcriptome analysis showed that, compared with the negative control group, the THP-1-Mp1p+ group had a total of 1 180 differentially expressed genes (DEGs), with 345 upregulated genes and 835 downregulated genes. GO enrichment analysis of DEGs showed that there were 135 differentially expressed genes, including 105 in biological processes (BP), 28 in cellular components (CC), and 2 in molecular functions (MF). The KEGG analysis results showed that the effect of Mp1p on THP-1 macrophages was highly correlated with the TNF pathway. The metabolomic analysis found that both the blank control group and the THP-1-Mp1p+ macrophage group achieved good separation between QC samples in both positive and negative ion modes. The threshold for significant differential metabolites was set at: VIP≥1 and T-test P<0.05, resulting in the identification of 488 differential metabolites, with 230 in the positive ion mode and 258 in the negative ion mode. Pathway enrichment analysis of the identified metabolites pointed to significant enrichment in metabolic pathways. The combined analysis confirmed that the tumor necrosis factor signaling pathway, interleukin-17 signaling pathway, and NF-kappaB signaling pathway were important metabolic pathways involved. Conclusions The virulence factor Mp1p may affect host macrophages by modulating the tumor necrosis factor signaling pathway, interleukin-17 signaling pathway, and NF-kappaB signaling pathway. The findings contribute to a better understanding of the mechanisms of action of Mp1p and may offer potential directions for the selection of relevant diagnostic and therapeutic targets in the future.

OBJECTIVETo study the feasibility of preparing a therapeutic lung cancer vaccine by transfecting dendritic cells (DCs) with adeno-associated virus vector carrying carcino-embryonic antigen gene (rAAV/CEA).

METHODSAdherent cells (monocytes) isolated from the peripheral blood of a healthy donor were infected with rAAV/CEA virus stock or pulsed with CEA peptide (control). The monocytes in both groups were induced into mature DCs with recombinant human GM-CSF, IL-4 and TNF-α. At day 7 of induction, the mature DCs were harvested and mixed with T lymphocytes. T cell proliferation stimulated by the DCs was assessed with (3)H-thymidine uptake, and the expression of IL-4, IFN-γ, CD8, CD4, CD25 and CD69 in cytotoxic T lymphocytes (CTL) was analyzed with flow cytometry. The cytotoxicity of the CTL against the target CEA-positive lung cancer A549 cells was tested by (51)Cr releasing assay.

RESULTSThe DCs transfected with rAAV/CEA strongly stimulated the proliferation of the T cell populations, and the induced CTL showed high expressions of CD8, CD69 and IFN-γ. The transfected DCs exhibited a high killing ability of CEA-positive lung cancer cells, and the killing showed a CEA antigen specificity and was limited by MHC I. These results suggested the ability of rAAV/CEA-transfected DCs in generating specific cellular immunity in vitro.

CONCLUSIONIt is feasible to prepare therapeutic lung cancer vaccines by transfecting DCs with rAAV/CEA.

Cancer Vaccines ; Carcinoembryonic Antigen ; genetics ; Cell Line ; Dendritic Cells ; immunology ; Dependovirus ; genetics ; Genetic Vectors ; Humans ; Monocytes ; immunology ; Transfection

Objective To analyze crossed cerebellar diaschisis(CCD) after subacute phase of spontaneous cerebral hemorrhage(SPSCH)and it's relevant factors with whole-brain CT perfusion(CTP) imaging. Methods Eighty-six patients diagnosed with unilateral SPSCH by CT were prospectively enrolled in our study from July 2015 to October 2016. Whole-brain CTP was performed in each patient.Cerebral blood flow(CBF), cerebral blood volume(CBV), mean transit time(MTT)and time-to-peak(TTP) inipsilateral and contralateral cerebellum were manually measured.The asymmetric indexs(AIs) were also calculated. Moreover, the volume of hematoma, the maximumarea of peri-hematomahypoperfusionin CBF and clinical factors(age, gender, time intervals from symptom onset)were analyzed,and NIHSS scores were used to evaluate the neurological status before patient admission, inspection, and discharge.CCD was rated positive when a unilateral supratentorial hematomawas appeared and an accompanying perfusion decrease was showed in the contralateral cerebellum on at least two sequential slices of CTP maps.All the individuals were divided into two groups including CCD-positive groupand CCD-negative group. The perfusion parameters (CBF, CBV, MTT, and TTP)between the contralateral and ipsilateral cerebellum were analyzed by the two-tailed paired t-test in CCD-positive group. The differences in the perfusion and clinical variables between the two groups were analyzed by the independent sample t-test and the Chi-squared test. Therelationships between the AI values and clinical or radiologic variables were assessed with Pearson correlation test. Results We found 35 CCD positive cases and 51 negative cases in the 86 patients.In CCD-positive groups, the perfusion values of cerebellumipsilateral and contralateral to the hematomawere as follows:CBF were (40.88±11.23) vs. (33.91±9.96) ml·100 g-1·min-1, CBV were (3.30±1.18) vs. (2.75±1.13) ml/100 g and TTP were (22.09±3.98) vs. (22.88±4.15) s, respectively, and there was statistical significance (t=10.231,8.223,-2.883,P<0.05).In CCD positive group, CBF, CBV, TTP, and MTT of the contralateral cerebellar hemisphere was changed in 35, 32, 26, and 16 cases,respectively.The AI value of CBF(AICBF)in CCD-positive group was (17.10±9.10)%, which was higher than that in the negative group (-0.95±17.01)%, there was statistical significance(t=6.367,P<0.05).The AI value of CBV(AICBV)was (17.43 ± 11.65)% in CCD-positive group, also significantly higher than that in negative group which was (1.55±21.06)%(t=4.477, P<0.05). No statistical difference(P>0.05)was found in hematoma location,hematoma volume, supratentorialhypoperfusion area and NIHSS scores(at admission, inspection)between CCD-positive and negative groups.The AICBF and AITTP showed linear correlation with time intervals in CCD-positive patients(P<0.05). Conclusions CCD is a common phenomenon in patients with SPSCH.Of all the perfusion parameters,CBF abnormalities are more common.The severity of CCD has a certain correlation with time intervals.There is no significant correlation between CCD and the clinical or radiological data(age, NIHSS scores,hematoma volume, and supratentorial hypoperfusion area).

Objective To study the feasibility of preparing a therapeutic lung cancer vaccine by transfecting dendritic cells (DCs) with adeno-associated virus vector carrying carcino-embryonic antigen gene (rAAV/CEA). Methods Adherent cells (monocytes) isolated from the peripheral blood of a healthy donor were infected with rAAV/CEA virus stock or pulsed with CEA peptide (control). The monocytes in both groups were induced into mature DCs with recombinant human GM-CSF, IL-4 and TNF-α. At day 7 of induction, the mature DCs were harvested and mixed with T lymphocytes. T cell proliferation stimulated by the DCs was assessed with 3H-thymidine uptake, and the expression of IL-4, IFN-γ, CD8, CD4, CD25 and CD69 in cytotoxic T lymphocytes (CTL) was analyzed with flow cytometry. The cytotoxicity of the CTL against the target CEA-positive lung cancer A549 cells was tested by 51Cr releasing assay. Results The DCs transfected with rAAV/CEA strongly stimulated the proliferation of the T cell populations, and the induced CTL showed high expressions of CD8, CD69 and IFN-γ. The transfected DCs exhibited a high killing ability of CEA-positive lung cancer cells, and the killing showed a CEA antigen specificity and was limited by MHC I. These results suggested the ability of rAAV/CEA-transfected DCs in generating specific cellular immunity in vitro. Conclusion It is feasible to prepare therapeutic lung cancer vaccines by transfecting DCs with rAAV/CEA.

Objective To study the feasibility of preparing a therapeutic lung cancer vaccine by transfecting dendritic cells (DCs) with adeno-associated virus vector carrying carcino-embryonic antigen gene (rAAV/CEA). Methods Adherent cells (monocytes) isolated from the peripheral blood of a healthy donor were infected with rAAV/CEA virus stock or pulsed with CEA peptide (control). The monocytes in both groups were induced into mature DCs with recombinant human GM-CSF, IL-4 and TNF-α. At day 7 of induction, the mature DCs were harvested and mixed with T lymphocytes. T cell proliferation stimulated by the DCs was assessed with 3H-thymidine uptake, and the expression of IL-4, IFN-γ, CD8, CD4, CD25 and CD69 in cytotoxic T lymphocytes (CTL) was analyzed with flow cytometry. The cytotoxicity of the CTL against the target CEA-positive lung cancer A549 cells was tested by 51Cr releasing assay. Results The DCs transfected with rAAV/CEA strongly stimulated the proliferation of the T cell populations, and the induced CTL showed high expressions of CD8, CD69 and IFN-γ. The transfected DCs exhibited a high killing ability of CEA-positive lung cancer cells, and the killing showed a CEA antigen specificity and was limited by MHC I. These results suggested the ability of rAAV/CEA-transfected DCs in generating specific cellular immunity in vitro. Conclusion It is feasible to prepare therapeutic lung cancer vaccines by transfecting DCs with rAAV/CEA.

AIM:To investigate the effects of Xiaozhong-Zhitong mixture(XZZT)on M2 polarization and Toll-like receptor 4(TLR4)/myeloid differentiation factor 88(MyD88)/nuclear factor-κB(NF-κB)signaling pathway in mouse microglia(BV2 cells).METHODS:The BV2 cells were divided into 5 groups:blank group,model group[lipo-polysaccharide(LPS)+hypoxia],TAK-242(resatorvid,a TLR4 inhibitor)group(LPS+hypoxia+TAK-242),XZZT group(LPS+hypoxia+XZZT),and TAK-242+XZZT group(LPS+hypoxia+TAK-242+XZZT).Flow cytometry was used to detect early apoptosis and cell cycle of BV2 cells,and immunofluorescence staining was employed to detect the positive expres-sion of M1-type marker inducible nitric oxide synthase(iNOS)and M2-type marker CD206.Western blot was utilized to detect the expression of TLR4/MyD88/NF-κB signaling pathway-related proteins,including TLR4,MyD88,NF-κB p65,phosphorylated p65(p-p65),phosphorylated transforming growth factor-β-activated kinase 1(p-TAK1),and phosphory-lated IκB kinase α/β(p-IKKα/β).RT-qPCR was used to detect the mRNA expression of interleukin-1β(IL-1β),IL-10,tumor necrosis factor-α(TNF-α),TLR4,MyD88,and NF-κB p65.RESULTS:Compared with model group,the rate of early apoptosis was significantly decreased in XZZT group(P<0.01),the percentage of cells arrested in the S phase was significantly increased(P<0.01),and the protein levels of TLR4,MyD88,NF-κB p65,p-IKKα/β,p-p65,and p-TAK1 were significantly decreased(P<0.05 or P<0.01).Additionally,IL-1β,TNF-α,TLR4,MyD88 and NF-κB p65 mRNA expression levels were significantly decreased(P<0.05 or P<0.01),while IL-10 mRNA expression was significantly in-creased(P<0.05).Compared with TAK-242 group,the average percentage of iNOS positive area was significantly de-creased,while CD206 was significantly increased in TAK-242+XZZT group(P<0.01).CONCLUSION:The XZZT has the effect of inducing M2 polarization of mouse microglia,and the mechanism may be linked to the inhibition of TLR4/MyD88/NF-κB signaling pathway.

BACKGROUND:In recent years,it has been found that some traditional Chinese medicine monomers can alleviate oxidative stress and apoptosis of the skin flap,promote vascular regeneration of the skin flap and prevent skin flap necrosis by activating autophagy. OBJECTIVE:To review the research progress of traditional Chinese medicine monomer regulating autophagy in preventing flap necrosis. METHODS:The Chinese and English key words were"traditional Chinese medicine(TCM),autophagy,skin flaps".The first author searched the relevant articles published in CNKI and PubMed databases from January 2010 to October 2022.A total of 196 articles were retrieved in the preliminary screening and then screened according to the inclusion and exclusion criteria.The quality assessment was conducted by reading the literature titles and abstracts.Finally,55 articles were summarized. RESULTS AND CONCLUSION:(1)The regulation of autophagy is mediated by AMPK/mTOR,PI3K/AKT and other signaling pathways.Activation of autophagy can alleviate the oxidative stress and apoptosis of the flap,promote the regeneration of blood vessels in the flap,and prevent flap necrosis.(2)Terpenoids(Betulinic acid,Andrographolide,Notoginseng Triterpenes,Catalpa),phenolic compounds(Resveratrol,Curcumin,Gastrodin),phenolic acids(Salvianolic acid B)and steroid compounds(Pseudoginsenoside F11)in traditional Chinese medicine monomers can alleviate oxidative stress and apoptosis of skin flap by regulating related signaling pathways to activate autophagy,promote skin flap angiogenesis and promote skin flap survival.(3)Studying the research progress of traditional Chinese medicine monomer to prevent flap necrosis by regulating autophagy can provide a reference and theoretical basis for traditional Chinese medicine to prevent flap necrosis and promote flap healing in the clinic.

BACKGROUND:Mesenchymal stem cells have great potential in the treatment of ischemia-reperfusion injury of skin flaps.However,their defects and the decline of their role in the treatment of ischemia-reperfusion injury of skin flaps restrict their wide application. OBJECTIVE:To review the strategies for improving the treatment of ischemia-reperfusion injury of skin flaps with mesenchymal stem cells,and provide a reference for its further theoretical research and clinical application. METHODS:Relevant documents included in CNKI,WanFang and PubMed were searched.The Chinese and English search terms were"mesenchymal stem cell,ischemia-reperfusion adjustment of skin flap,mesenchymal stem cells,stem cells,skin flap,ischemia-reperfusion injury,pretreatment,gene modification,biomaterial packaging,joint application".The relevant documents since 2007 were retrieved,and the documents with little relationship between the research content and the article theme,poor quality and outdated content were eliminated through reading the article,and finally 75 documents were included for summary. RESULTS AND CONCLUSION:(1)Mesenchymal stem cells can inhibit inflammatory reactions,resist oxidative stress and induce angiogenesis,which has great potential in the treatment of skin flap ischemia-reperfusion injury.(2)Although mesenchymal stem cells have shown great potential in the treatment of skin flap ischemia-reperfusion injury,their shortcomings in treatment have limited their widespread clinical application.Through pre-treatment(cytokines,hypoxia,drugs,and other pre-treatment mesenchymal stem cells),gene-modified mesenchymal stem cells,biomaterial encapsulation of mesenchymal stem cells,as well as the combined use of mesenchymal stem cells and other drugs or therapeutic methods,can not only overcome the shortcomings of mesenchymal stem cells in treatment,but also improve their therapeutic effectiveness in skin flap ischemia-reperfusion injury.(3)Therefore,further improving the effectiveness of mesenchymal stem cells in treating skin flap ischemia-reperfusion injury and exploring its therapeutic potential are of great significance for the research of mesenchymal stem cells and the treatment of skin flap ischemia-reperfusion injury.