Objective To study inflammatory reaction induced by N-protein of severe acute respiratory syndrome(SARS)-coronavirus(CoV)in human alveolar typeⅡepithelial cell(A549). Methods Effects on growth of A549 cell by N-protein of SARS-CoV:activity of A549 cells was determined by thiazylyl blue colorimetry assay at 24,48,72 and 96 h,respectively.Effects on cyto- kine production by A549 cells exposed to N-protein of SARS-CoV:interleukin(IL)-6,IL-10 and transforming growth factor-?1(TGF-?1)concentration in culture supernatant were determined by enzyme-linked immunosorbent assay(ELISA).Effects on mRNA expression of cytokine of A549 cells and matrix metalloproteinases-9(MMP-9)exposed to N-protein of SARS-CoV:total RNA of A549 cells was extracted using Rneasy mini kit;RT-PCR was employed to measure the mRNA expression of IL-6,IL-10,TGF-?1 and MMP-9 semiquantitatively.Results Different concentrations of N-protein could all inhibit the growth of A549 cells(after 48 h)and the inhibition by 20?g/mL pro- tein was the strongest.Compared with the control group(0.737?0.024,0.968?0.007),the A val- ues of experimental groups at 72 h and 96 h(0.672?0.027,0.799?0.092)decreased obviously (P

OBJECTIVETo investigate MRI findings of osteochondral lesions in the talus;to evaluate the value of MRI in diagnosing and determining the stage of osteochondral lesions;to analyze the follow up clinical value of MRI in osteochondral transplantation of autologous bone.

METHODSA total of 79 patients from February 2013 to March 2015 had been retrospectively analyzed. All the patients were treated in our hospital. The ankle arthroscopy results were used as the reference standard, and the accuracy of MRI in diagnosis and Hepple staging had been investigated. Fifteen patients with cartilage transplantation of autologous bone were followed up with MRI examination and evaluation of cartilage repair score(MOCART) after one year. The values of MRI in the postoperative follow up were analyzed.

RESULTSHepple staging of 79 patients was shown as follows:7 cases of stage I, 12 cases of stage II, 24 cases of stage III, 16 cases of stage IV, and 20 cases of stage V. Ankle arthroscopy grading of 59 patients in this group(in addition of 20 cases of stage V):2 cases of grade A, 2 cases of grade B, 4 cases of grade C, 14 cases of grade D, 22 cases of grade E, and 15 cases of grade F. The accuracy rate of MRI in determining Hepple V was set at 100%, and Hepple stage I corresponds to the arthroscopic A, B, C stage, stage II corresponds to D stage, stage III corresponds to E stage, stage IV corresponds to F stage. The accuracy rate of MRI in determining Hepple stage I to IV was 87.5%, 85.7%, 95.4% and 93.3% respectively. After cartilage transplantation of autologous bone, MRI of 15 patients showed cartilage surface in transplanted area was smooth, bone healed well, and the surrounding edema disappeared. The MOCART was 30 to 80 scores with an average score 59.0±15.6;9 cases of these 15 patients were(9/15, 60%) higher than 60 score.

CONCLUSIONSMRI plays a significant role in clinical diagnosis and staging of the talus osteochondral injury. As a method of long term follow up after cartilage transplantation, MRI can well evaluate the rapair of the postoperative osteochondral injury.

Objective:To analyze the expression of LETM2 in KYSE150 and ECA109 cell lines and its effect on the proliferation, migra-tion, and invasion of esophageal squamous cell carcinoma (ESCC). Methods:The expression level of the LETM2 protein in 90 paired hu-man ESCC tissues and matched adjacent normal tissues was determined through immunohistochemistry. The expression level of LETM2 in ESCC cell lines was detected by real-time PCR and Western blot. The expression levels of LETM2 in KYSE150 and ECA109 cell lines were knocked down using lentivirus. MTT assays were performed to examine the effect of LETM2 on the proliferation of ESCC cells. Colony formation assay was used to detect the colony formation ability. Flow cytometry was performed to analyze the cell cycle. The effect of LETM2 depletion on the migration and invasion of ESCC cells was determined by Transwell assay. Results:LETM2 expres-sion was frequently upregulated in the ESCC tissues than in the adjacent normal tissues. The suppressed exogenous expression of LETM2 led to the inhibition of cell proliferation and colony formation. However, cell migration and invasion were not affected. The re-sults on the cell cycle distribution revealed that LETM2 knockdown acts as a negative regulator of the cell cycle at the G1 to S phase transition. Conclusion:LETM2 acts as a tumor-driven gene in the development and progression of ESCC. This finding suggests that LETM2 can be used as an efficient prognosis biomarker and a potential therapeutic target for ESCC.

OBJECTIVETo investigate the methods of culturing and identifying mouse myeloid semimature dendritic cell (smDC) in vitro.

METHODSMyeloid monocytes derived from 6-week-old C57 BL/6 mice were cultured in RPMI-1640 medium containing 10% fetal bovine serum, 2 ng/ml recombinant murine granulocyte macrophage-colony stimulating factor (GM-CSF), and 20 ng/ml recombinant murine interleukin (IL)-4 for 9 days. Then cells were incubated with 40 ng/ml tumor necrosis factor-alpha (TNF-alpha) for 24 hours to obtain smDC. Meanwhile, smDC was differentiated into mature dendritic cell (mDC) or immature dendritic cell (iDC) by treatment with 1 micro/m1 lipopolysaccharide (LPS) or without LPS. The morphological features of smDC were assayed by inverted microscopy and scanning electron microscopy. Surface markers such as CD11c, CD4O, CD8O, CD86, and MHC-II were tested by flow cytometry. IL-1beta, IL-6, IL-12, and IL-10 in the supernatant were tested by ELISA. The activation of allogene lymphocyte (BALB/c mice) stimulated by C57BL/6 myeloid smDC in mixed lymphocyte reaction was examined by Cell Counting Kit-8 in vitro.

RESULTSThe shape of smDC was round or oval-shaped, and the diameter of smDC was about 15 microm. The length of smDC dendrite was between 5 to 10 microm. smDC, iDC, and mDC all expressed high level of CD11 c. The expressions of MHC-II, CD40, CD80, and CD86 on smDC were higher than those of iDC and lower than those of mDC. IL-1beta, IL-6, and IL-12 secretion of smDC was significantly lower than that of mDC (P < 0.01), and IL-12 was significantly lower than that of iDC (P < 0.05), while no significant difference of IL-1beta and IL-6 secretion was found between smDC and iDC (P > 0.05). Furthermore, IL-10 secretion was not significantly different among these three kinds of DCs (P > 0.05). The effect of allogene lymphocytes activation on smDC was significantly lower than that of mDC and positive control (P < 0.01), but had no significant difference when compared with that of iDC and negative control (P > 0.05).

CONCLUSIONSsmDC may be a relatively independent dendritic cell sub-population in terms of function and morphology. It is a feasible way to induce myeloid monocytes to differentiate into smDC using GM-CSF, IL-4, and TNF-alpha in vitro.

Animals ; Cell Culture Techniques ; Cell Differentiation ; Cells, Cultured ; Cytokines ; immunology ; Dendritic Cells ; cytology ; immunology ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Inbred C57BL ; Monocytes ; cytology ; immunology

AIMTo study the effect of Angelica sinsensis polysaccharides on lymphocyte proliferation and induction of IFN-gamma.

METHODSAngelica sinensis polysaccharides(AP) were separated into AP-I, AP-II, AP-III and AP-IV by alcohol deposition with different concentration. The radioactivities of [3H]-TdR uptake by lymphocyte were used to determine the ability of lymphocyte. The bioactivity of IFN-gamma was measured by violet crystalline dying.

RESULTSAP-IV was found to be composed of Ara and Glu in the ratio of 0.99:6.47, the molecular weight was estimated to be 5,600. AP-I and AP-II 100 mg.kg-1 i.p. were found to significantly augment mice splenocyte proliferation, release IFN-gamma and increase IFN-gamma bioactivity. 50 micrograms.mL-1 AP-I, AP-II and AP-III were shown to enhance the proliferative response of the mouse spleen lymphocytes in vitro.

CONCLUSIONAP-I and AP-II showed higher immunoactivity than AP-III, AP-IV had no effect.

Angelica sinensis ; chemistry ; Animals ; Cell Division ; drug effects ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Immunity ; drug effects ; Interferon-gamma ; biosynthesis ; Lymphocytes ; cytology ; drug effects ; immunology ; Male ; Mice ; Mice, Inbred BALB C ; Plants, Medicinal ; chemistry ; Polysaccharides ; isolation & purification ; pharmacology ; Random Allocation ; Spleen ; cytology ; Tumor Cells, Cultured

With the diversion rate of ginkgolide A, B, K as comprehensive evaluation indexes, the amount of activated carbon, ad- sorption time, mix rate, and adsorption temperature were selected as factors, orthogonal design which based on the evaluation method of information entropy was used to optimize activated carbon adsorption technology of ginkgo diterpene lactones meglumine injection. Opti- mized adsorption conditions were as follows: adsorbed 30 min with 0.2% activated carbon in 25 °C, 40 r ·min⁻¹, validation test re- sult display. The optimum extraction condition was stable and feasible, it will provide a basis for ginkgo diterpene lactone meglumine injection' activated carbon adsorption process.
Adsorption ; Charcoal ; chemistry ; Chemistry, Pharmaceutical ; instrumentation ; methods ; Diterpenes ; chemistry ; isolation & purification ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Ginkgo biloba ; chemistry ; Lactones ; chemistry ; isolation & purification

Objective:To investigate the association between metabolically healthy obesity(MHO) and atherosclerosis risk among Chinese community population aged 40 or older.Methods:A total of 9 525 participants without cardiovascular diseases (3 621 men and 5 904 women) from Jiading community in Shanghai were enrolled to complete questionnaires, undergo extensive physical examination including brachial-ankle pulse wave velocity (baPWV) and blood pressure (BP) assessment, and laboratory screening. According to body mass index (BMI) and metabolic status, these participants were categorized into 4 groups including metabolically healthy non-obese (MHNO), metabolically unhealthy non-obese (MUNO), MHO, and metabolically unhealthy obese (MUO). High baPWV was defined as baPWV>1 400 mm/s, and high pulse pressure (PP) was defined as PP above fourth quartile of the population. Multivariate logistic regression model was conducted to explore the relationship between MHO and high baPWV as well as high PP after adjusting for confounders. Results:After multivariable adjustment, such as sex, age, current smoking, current drinking, and education, logistic regression analysis showed that MHO was significantly correlated with high baPWV ( OR=1.18, 95% CI 1.02-1.37) and high PP ( OR=1.72, 95% CI 1.43-2.08) in comparison with MHNO. Otherwise, both MUNO and MUO subjects were at higher risk for suffering from high baPWV (MUNO: OR=3.02, 95% CI 2.60-3.50; MUO: OR=3.26, 95% CI 2.87-3.70) and high PP (MUNO: OR=2.56, 95% CI 2.17-3.02; MUO: OR=3.49, 95% CI 3.01-4.06). Conclusion:On the basis of Chinese community population, there was a pronounced correlation between the MHO phenotype and the increased risk of developing atherosclerosis.

Coronavirus is human-to-human transmissible and often leads to nosocomial outbreak in the early stage, which poses a great threat to healthcare workers.Due to confined space, crowed population and some aerosol-generating medical procedures, hospital is vulnerable to coronavirus transmission and nosocomial infection.We review the mechanism and risk factors of coronavirus aerosol in hospital and summarize the evidence of aerosol transmission of three coronaviruses and the impact of air flow.Furthermore, we evaluate the effectiveness of personal protection in the above-mentioned scenarios for the prevention and control of nosocomial infection.

With the increasing application of immune checkpoint inhibitors(ICI)in anti-tumor therapy,ICI related infections are often neglected.Mycobacterium tuberculosis(MTB)is also a common pathogen.We reported a case of ICI related pulmonary tuberculosis from Zhongshan Hospital,Fudan University.Meanwhile,18 cases of ICI related tuberculosis infection were collected through literature search,and the characteristics of ICI related tuberculosis were analyzed to improve the understanding in clinic practice.All the cases were confirmed TB including 15 cases of pulmonary tuberculosis(1 case with complication of intestinal tuberculosis)and 4 cases of extra-pulmonary tuberculosis(1 case of disseminated tuberculosis,bone tuberculosis,tuberculous pericarditis and tuberculous pleurisy,respectively).The chest CT characteristics of pulmonary tuberculosis mainly included centrilobular nodules,ground glass nodules,empty lesions,patchy shadows,consolidation and large infiltration.Eighteen cases started anti-tuberculosis treatment,while 4 cases continued ICI treatment.Three cases suspended ICI(2 cases had remission after reuse)and 11 cases stopped ICI,and 1 case was not mentioned.ICI related tuberculosis may be a direct complication of tumor immunotherapy.It is necessary to screen tuberculosis infection and exclude active tuberculosis before immunotherapy.If there are suspected symptoms such as fever,cough and sputum during ICI treatment,active tuberculosis should be taken into account.

In order to explore the specificity of complement C5 in the postmortem diagnosis of myocardial infarction, changes of C5 staining in normal, infarcted and other non-infarcted myocardia with direct or indirect myocardial injuries (myocarditis, mechanical asphyxia, electrocution, hemorrhagic shock, cardiac contusion and organophosphate poisoning) were studied with immunohistochemistry and image analysis. The results showed that positive C5 staining could be observed in groups of myocardial infarction and myocarditis, but not in groups of mechanical asphyxia, electrocution, hemorrhagic shock, cardiac contusion, and organophosphate poisoning. It is indicated that positive reaction of C5 could only be affected by myocarditis, which means that it was more specific for the diagnosis of myocardial infarction.
Adolescent ; Adult ; Complement C5/analysis* ; Female ; Forensic Medicine ; Humans ; Male ; Myocardial Infarction/diagnosis* ; Myocardium/chemistry* ; Sensitivity and Specificity