BACKGROUND:Pulmonary aspergilosis is a disease caused by pulmonary fungal infection. Its diagnosis and treatment is usualy delayed because of nonspecific clinical symptoms, physicial sign and imaging changes as wel as uncertainties of histological and bacterial findings. Therefore, it is necessary to establish mouse models of invasive pulmonary aspergilosis to investigate the underlying pathological mechanism and novel therapeutic methods. OBJECTIVE: To establish mouse models of invasive pulmonary aspergilosis, detect the expression ofγ-interferon, Tol-like receptor 2 and Tol-like receptor 4, and discuss the mechanism of action underlying invasive pulmonary aspergilosis. METHODS:Seventy-five female BALB/c mice of clean grade, aged 6-8 weeks, were randomly and evenly divided into five groups: blank control group (group A), immunosuppressive model group treated with high concentrations of Aspergilus fumigatus spore suspension (group B), normal infection group treated with high concentration of Aspergilus fumigatus spore suspension (group C), immunosuppressive model group treated with low concentration of Aspergilus fumigatus spore suspension (group D), normal infection group treated with low concentration of Aspergilus fumigatus spore suspension (group E). First, mice in the groups B and D were intraperitonealy injected with cyclophosphamide to establish immunosuppressive models. The mice in the groups D, E (108 cfu/mL) and groups B, C (109 cfu/mL) were treated with 12 mL Aspergilus fumigatus spore suspension through the use of nebulizer. Mice in the group A were treated identicaly with sterile PBS. At 1, 3, 5 days of infection, the pathological change of lung tissue was observed, the mass concentration of γ-interferon in bronchoalveolar lavage fluid and the expression levels of γ-interferon mRNA and Tol-like receptor 2 and Tol-like receptor 4 mRNA and protein in the lung tissue were determined. RESULTS AND CONCLUSION:Abscess, spores and very severe bleeding and congestion, widenened alveolar septum and tracheal epithelial cel shedding and necrosis were observed in the mouse lung tissue in the group B. At 5 days of infection, the mass concentration of γ-interferon in bronchoalveolar lavage fluid and the expression ofγ-interferon mRNA in the lung tissue in the group B were significantly decreased compared with the group A (P < 0.05). Tol-like receptor 2 expression was strongly positive in the group B. Tol-like receptor 2 expression in the group C was significantly lower than that in the group B (P< 0.05). Tol-like receptor 4 expression was positive in the groups B and C, and its expression in the group C was significantly greater than in the group B (P < 0.05). The expression of Tol-like receptor 2, 4 mRNA in the mouse lung tissue of group B was significantly increased at 1, 3, 5 days of infection (P < 0.05). These results suggest that atomizing high concentration of aspergilus fumigatus spore suspension to immunosuppressive mice can establish stable invasive pulmonary aspergilosis models with typical pathological features. The infection of aspergilus fumigatus can activate tol-like receptor 2, 4 at the same time, and the pathological mechanism is closely related to organism’s immune defense function.

Objective To probe into impacts of safflor yellow injection on rats with acute myocardial infarction and discuss its mechanism. Methods Forty Sprague-Dawley rats were divided into sham operation group ( 8 rats ) and operation groups(32 rats).The left anterior descending branch of coronary artery was ligated to establish an animal model with acute myocardial infarction.The rats with successful modeling were randomly divided into model control group, 3-, 7- and 14-days safflor yellow group.In the treatment group, the rats were injected with safflor yellow injection(2.5 mg.kg-1.d-1)for 3, 7 and 14 days, respectively.Rats in sham operation and model control group were injected 0.9% sodium chloride solution.Their weights were measured 24 h after the last drug administration, their hearts were removed, and the serum was obtained through centrifugation.The enzymic method was used to measure the lactate dehydrogenase(LDH)and creatine kinase(CK)contents of serum.Hematoxylin eosiny(HE)staining method was used to observe the change of myocardial cells.The chloride three phenyl-tetrazole( TTC) staining method was used to measure the area of myocardial infarction in the rats. The real-time fluorescent quantitative PCR method was used to measure the expression of VEGF and HIF mRNA at the marginal zone of myocardial infarction. Results The levels of LDH and CK were(106.12±35.52),(452.84±60.38)mmol.L-1 in sham operation group, and(385.66±137.58),(2 111.00±1 250.80)mmol.L-1 in model control group.LDH in 3-, 7-, 14-days treatment groups was (249.66±51.86),(104.33±51.08),(110.33±26.76)mmol.L-1, while CK was(1 713.00±584.74),(1 177.66±980.18), (421.33±54.60)mmol.L-1.LDH and CK levels of the 14-days treatment group were significantly lower than those in the model control group(both P<0.05).The area of myocardial infarction was (0.00±0.00)% in sham operation group,(13.12±6.69)% in the model control group, and(8.11±3.45)%,(7.01±2.98)%,(3.44±1.17)% in the 3-, 7-, 14-days treatment groups.The area of myocardial infarction in the 14-days treatment group was significantly lower than that of the model control group(P<0.05).The expression of VEGF and HIF mRNA was 1.99±0.27, 6.21±1.35 in the sham operation group, 1.03±0.15, 1.78±0.57 in the model control group.The expression of HIF mRNA in the 3-, 7-, 14-days treatment groups was 0.50±0.12, 1.23±0.24, 2.20± 0.32, and the expression of VEGF mRNA in the 3-, 7-, 14-days treatment groups was 0.43±1.27, 2.67±0.83, 5.78±1.23. Conclusion The safflor yellow injection has therapeutical effects on the rats with acute myocardial infarction, and the effects will become significant after 14 day.It can reduce the area of myocardial infarction and LDH and CK content at the drug administration groups, which might be related to the increase of the expression of VEGF and HIF mRNA.

Objective To reestablish neuroimmunoregulation network and its function.Methods Whole-some gene array was used to analyze the gene expression involved in neuroimmunoregulation function of the lateral hypothalamic(LH) in rats at different immunizational time.The function trees of different genes were analysed with the PathWay Miner public database.Results 632 genes were differentially expressed: including 374 of 2-day immunized group,62 of 4-day immunized group and 196 of 6-day immunized group.Function signal pathway analysis for 398 up-regulated genes showed 27 genes were involved in 31 cell functional signal conduction pathways,including the well-known signal conduction pathways of synaptic reconstruction.Conclusion Synapses reconstruction appears to be the important pattern of functional reestablishment of neuroimmunoregulation network.

Objective:To study 3-dimensional facial soft tissue reference values of Chinese Han nationality adults with normal occlusion and explore the facial morphology differences between men and women.Methods:3dMD images from 30 men and 32 women of Han nationality,aged 18-35 years,with normal occlusion were analyzed and compared between the men and women on 19 linear distances,8 angular measurements and 10 linear distance ratios.Results:Linear distances concerning the width and height of facial and organic contours were larger in men(P<0.05),while the ratio of facial height to width were larger in women(P<0.05);the nasofrontal angle and nasal prominence angle were both smaller(P<0.05) but nasofacial angle were larger(P<0.05) in men than in women.Conclusion:The facial and organic contours are larger in men than in women.Men have wide-elliptical face while women have thin-elliptical one.The nose of men is more prominent than women.

Objective:To observe the changes of macular microvessels in patients with retinal vein occlusion (RVO) and macular edema (ME) after intravitreal injection of aflibercept (IVA), and analyze its correlation with best corrected visual acuity (BCVA).Methods:A retrospective case study. Thirty patients (30 eyes) with monocular RVO with ME (RVO-ME) who were diagnosed in the clinical examination of Tianjin Eye Hospital from April 2019 to February 2020 were included in the study. Among them, there were 12 males (12 eyes) and 18 females(18 eyes); the average age was 54.30±13.17 years. The average course of disease was3.43±1.97 months. Both eyes were examined by BCVA and optical coherence tomography (OCTA). The on-demand injection was adopted after the first injection in IVA treatment regimen. The macular area 6 mm×6 mm in both eyes was scanned with an OCTA instrument, and the area of the foveal avascular area (FAZ), FAZ circumference (PERIM), and out-of-roundness were measured at baseline and 1, 3, and 6 months after treatment. Index (AI), blood flow density within 300 μm width of FAZ (FD-300), foveal retinal thickness (CMT), superficial retinal capillary plexus (SCP), deep retinal capillary plexus (DCP) blood flow density. The paired t test was used to compare the quantitative parameters of the affected eye and the contralateral healthy eye at baseline; the changes of the quantitative parameters at baseline and 1, 3, and 6 months after treatment were analyzed by repeated measures analysis of variance. Pearson correlation analysis was used to analyze the correlation between BCVA, retinal perfusion, and macular blood supply parameters at 6 months after IVA treatment. Results:At baseline, compared with the contralateral healthy eye, the FAZ area ( t=-4.091), PERIM ( t=-5.098) and AI ( t=-9.093) of the RVO-ME eye were enlarged, and FD-300 ( t=7.237) and overall SCP and DCP blood flow density ( t=8.735, 9.897) decreased, the difference was statistically significant ( P<0.001). Six months after treatment, the BCVA of RVO-ME eyes was significantly increased, CMT decreased, FAZ area expanded, and AI decreased ( t=8.566, 16.739, -6.469, 9.719; P<0.001), the difference was statistically significant. There was no significant change in the blood flow density of FD-300 and overall SCP and DCP, and the difference was not statistically significant ( t=1.017, 1.197, 0.987; P>0.05). Compared with baseline, the FAZ area of RVO-ME eyes gradually expanded at 3 and 6 months after treatment, and the difference was statistically significant ( F=21.979, P<0.001). Correlation analysis results showed that BCVA at 6 months after treatment was positively correlated with the overall SCP and DCP blood flow density at baseline and 6 months after treatment ( r=-0.538, -0.484, -0.879, -0.854; P<0.05). There was a negative correlation with the area of FAZ 6 months after treatment ( r=0.544, P=0.001). The number of ME recurrences was negatively correlated with BCVA and overall SCP and DCP blood flow density 6 months after treatment ( r=0.604, -0.462, -0.528; P<0.05), it was positively correlated with FAZ area ( r=0.379, P=0.043). Conclusion:Within 6 months of IVA treatment in RVO-ME eyes, ME is significantly reduced and visual acuity is improved; SCP blood flow density decreases, and FAZ area expands.

PURPOSE: Accumulating evidence has shown that dysregulation of microRNA-191 (miR-191) is closely associated with tumorigenesis and progression in a wide range of cancers. This study aimed to explore the potential role of miR-191 in esophageal squamous cell carcinoma (ESCC). MATERIALS AND METHODS: miR-191 expression was assessed in 93 ESCC tissue specimens by real-time polymerase chain reaction, and survival analysis was performed via Kaplan-Meier and Cox regression analyses. 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide, plate colony-forming, BrdU, and Transwell assays were conducted to observe the effect of miR-191 on ESCC proliferation and invasion. Luciferase reporter and western blot assays were taken to identify target genes of miR-191. RESULTS: miR-191 was overexpressed in 93 cases of ESCC, compared with adjacent normal tissues, and miR-191 expression was significantly related to differentiation, depth of invasion, TNM stage, lymph node metastasis, and distant metastasis of tumor. Kaplan-Meier and Cox regression analyses demonstrated that overexpression of miR-191 was an independent and significant predictor of ESCC prognosis. Both gain-of-function and loss-of-function experiments showed that miR-191 promoted ESCC cell proliferation and invasion activities in vitro. Early growth response 1 (EGR1), a tumor suppressor, was predicted as a direct target of miR-191. Luciferase reporter and western blot assays proved that miR-191 reduced EGR1 expression by directly binding its 3' untranslated region. Moreover, EGR1 knockdown by siRNA enhanced ESCC cell growth and invasion. CONCLUSION: Our findings provide specific biological roles of miR-191 in ESCC survival and progression. Targeting the novel miR-191/EGR1 axis represents a potential new therapeutic way to block ESCC development.
3' Untranslated Regions ; Blotting, Western ; Bromodeoxyuridine ; Carcinogenesis ; Carcinoma, Squamous Cell* ; Cell Proliferation ; Epithelial Cells* ; In Vitro Techniques ; Luciferases ; Lymph Nodes ; Neoplasm Metastasis ; Prognosis* ; Real-Time Polymerase Chain Reaction ; RNA, Small Interfering

Objective:To explore the influencing and prognosis factors of emphysematous urinary tract infection (EUTI).Methods:The baseline clinical data of the patients admitted to Shandong University Qilu Hospital (Qingdao) from December 2013 to June 2020 and diagnosed with EUTI were analyzed retrospectively. The patients with non-EUTI (NEUTI) during the same period were selected as the control group. The baseline characteristics between the two groups were compared. Logistic regression analysis method was used to analyze the influencing factors of EUTI.Results:(1) 24 EUTI patients and 53 NEUTI patients were included in the present study. Compared with the NEUTI group, the hemoglobin level was lower ( t=-5.245, P<0.001) and the levels of blood urine nitrogen ( Z=-4.361, P<0.001), serum creatinine (Scr, Z=-4.543, P<0.001), blood glucose ( Z=-2.608, P=0.009), and triacylglycerol ( Z=-2.408, P=0.016) were higher in the EUTI group. The proportions of diabetes mellitus ( χ2=13.453, P<0.001) and chronic kidney disease ( χ2=17.936, P<0.001) in the EUTI group were higher than those in the NEUTI group. Increasing Scr was the risk factor of EUTI in patients with urinary tract infection ( OR=1.011, 95% CI 1.001-1.020, P=0.025). (2) Escherichia coli ( E.coli, 14 cases, 58.3%) was the most common causative organism. The other causative organisms included Klebsiella pneumoniae (2 cases, 8.3%), Enterococcus faecium (1 case, 4.2%), Pantoea (1 case, 4.2%), and mixed bacteria of E.coli and Enterococcus faecium (1 case, 4.2%). Ten cases of E.coli were extended-spectrum β-lactamases (ESBL)-positive. (3) Of the 24 patients with EUTI, 4 patients had adverse outcomes. The length of stay ( Z=-2.457, P=0.014), blood urea nitrogen ( t=2.432, P=0.024), shock ( P=0.002), autoimmune disease ( P=0.022), and white blood cell count ( Z=-2.091, P=0.036) were statistically different between good prognosis group ( n=20) and poor prognosis group ( n=4). However, logistic regression analysis results showed that neither was the influencing factor of poor prognosis of EUTI. Conclusions:The elevated Scr level is the independent influencing factor of EUTI among urinary infection patients. E.coli is the most common pathogenic bacteria, and ESBL-positive bacteria are common.

Objective: To explore the significance of next-generation sequencing for the screening of high-risk hereditary gastrointesti-nal cancer patients and the value of high-risk factors in screening. Methods: Twenty-five hereditary high-risk gastrointestinal cancer pa-tients from March 2016 to April 2016 in Peking University Cancer Hospital were enrolled. They received detection of 42 hereditary can-cer syndrome related genes by next-generation sequencing. Results: Out of 25 patients enrolled, 24% (6/25) patients had pathological germline mutations. The expression of mismatch repair protein was absent in 50% (3/6) patients. There were 83% (5/6) patients with family history of malignant tumors and were diagnosed when younger than 50 years. Six patients had hereditary cancer syndrome re-lated gene mutation, 1 patient had MYH gene missense mutation, 1 patient had APC gene deletion mutation, 4 patients had heredi-tary colorectal cancer related gene mutation, including MLH1, MLH3, and TGFBR2 germline missense mutations as well as MSH6 non-sense mutation. Conclusions: Out of 25 patients with high-risk factors of hereditary gastrointestinal cancer, 6 (24%) had pathological germline mutations. Given the high frequency and wide spectrum of mutations, the application of next-generation sequencing for screening of hereditary high-risk gastrointestinal cancer patients has the clinical value for improving the positive rate of diagnosis.

Objective:To establish a method for determintation of chlorogenic acid and linarin in Yejuhua granules by HPLC.Methods:We applied HPLC methods. The Kromasil 100-5 C18 column (250 mm×4.6 mm,5 μm) was used, the mobile phase was acetonitrile-0.4%H 3PO 4 solution (gradient elution), the flow rate was 1.0 ml/min, the dection wavelenghth was 334 nm and the column temperture was 32 ℃. Results:Chlorogenic acid and buddleoside had good linearity in the ranges of 0.30-1.50 μg ( r2=0.999 1) and 0.12-0.62 μg ( r2=0.999 8), respectively. The average recoveries were 99.70% and 96.67%, with RSD<2%, respectively. Conclusion:The method is simple, rapid, reliable, efficient, and can be used for determination of chlorogenic acid and buddleoside in Yejuhua Granules.

Objective It has been confirmed that Epstein-Barr virus ( EBV) is associated with the occurrence and development of the nasopharyngeal carcinoma ( NPC ) . We investigated the clinical significance of plasma concentrations of EBV-DNA in patients with NPC. Methods Since October,2013 to December,2016,471 patients were analyzed. The significantly associated between EBV-DNA before treatment and staging, tumor burden was analyzed. The survival rate of EBV-DNA before and after treatment was calculated. Results The median copies of pretreatment plasma EBV-DNA in patients is 137 copies,( range 0-494000) ,which is correlated with T stage,N stage,M stage,clinical stage and tumor burden load and that is statistically significant. Overall survival ( OS,P=0. 007) ,progression-free survival ( PFS,P=0. 011) and distant metastasis-free survival ( DMFS,P=0. 003) were significantly lower among patients with pretreatment plasma EBV-DNA more than 1300 copies/ml. Patients with detectable plasma EBV-DNA had significantly worse OS (P=0. 016),PFS (P=0. 000) and DMFS (P=0. 000) than patients with undetectable EBV-DNA after treatment. Cox multivariate analyze suggests that T stage and EBV-DNA after treatment were independent prognostic factors for OS,however the plasma EBV-DNA after treatment ( P=0. 006,0. 001) and N stage ( P=0. 037,0. 017) were independent prognostic factors for PFS and DMFS. Conclusions The plasma EBV-DNA level was significantly correlated with staging and tumor load before treatment in patients with NPC,and the prognosis of patients with higher copies before treatment could be worse. The plasma EBV-DNA after treatment is predictive for OS,PFS and DMFS.