Objective To understand the situation of off-label use of sildenafil, so as to provide reference for developing off-label drug use policy. Methods A stratified random sampling method was used to collect prescription data of sildenafil in Guangdong Women and Children Hospital in 2015. The off-label drug use situation was analyzed according to drug labels. The situations of drug use in all departments were summarized, and off-label drug use occurrences were analyzed.Results A total of 1044 prescriptions with sildenafil were collected and analyzed to discover the occurrence of off-label use was up to 69. 54%. The top 3 departments of off-label use of sildenafil were assisted reproductive center, heart center and neonatal intensive care unit (NICU). The top 3 types of off-label use were non-indication(50. 35%), over drug population(31. 76%) and over drug administration(17. 89%). The top 3 diseases for non-indications were infertility, pulmonary hypertension and congenital heart disease. Conclusion Sildenafil off-label use was common in Guangdong Women and Children Hospital in 2015, especially in assisted reproductive center, heart center, and NICU. The result suggests that more clinical studies about sildenafil on cardiovascular system are expected to provide more evidence for drug use, so as to ensure the safety of drug use in special populations and avoid potential medical risk.

AIM: To observe the effects of dexamethasone on the expression of toll-like receptor 4(TLR4)and the activation of NF-?B of dendritic cells(DC),and investigate the mechanism of dexamethasone suppressing differentiation,maturation,and function of DC.METHODS: DC generated from C57BL/6 murine bone marrow cells were induced by GM-CSF and IL-4.Dexamethasone was added in the DC's culture system,the cells were analyzed by flow cytometry(FCM) to determine the expression of TLR4 & CD11c,and by electrophoretic mobility shift assay(EMSA) to verify the activation of transcription factor NF-?B of DC.RESULTS: Dexamethasone could suppress the expression of CD11c and the activation of NF-?B with a dose-effect relationship in DC,but it could not suppress the expression of TLR4 of DC.CONCLUSION: The mechanism of dexamethasone involved in inhibition of differentiation,maturation,and function of DC is perhaps related to suppression of NF-?B activation.

Objective To detect the serum IgG4 and autoantibodies levels in patients with orbital disease of unknown reasons,and to investigate their values in patients with orbital disease.Methods A total of 366 patients with orbital disease of unknown reasons recruited in the Department of Ophthalmology,Beijing Tongren Hospital Affiliated to Capital Medical University from October 2013 to October 2016 were retrospectively enrolled as orbital disease group,and 266 patients with autoimmune disease in the same period from the Department of Rheumatology of the hospital were selected as controls.The serum IgG4 was detected by rate scattering method,antinuclear antibody(ANA),anti-double-stranded DNA(dsDNA)antibody as well as anti-extractable nuclear antigen(ENA)antibody were measured by indirect immunofluorescence assay,and anti-neutrophil cytoplasmic antibody(ANCA)was detected by enzyme linked immunosorbent assay,all of which were compared between the orbital disease patients and the controls using chi-square test.Results The positive rate of the serum IgG4 in the patients with orbital disease was 36.1％(132/366),obviously higher than that in the controls(27.1％,72/266),the difference being statistically significant(x2 =5.705,P=0.017).And the positive rate of serum IgG4≥1 350 mg/L(29.0％,106/366)in the patients with orbital disease was higher than that in the controls(21.8％,58/266; x2 =4.107,P=0.043).The positive rate of ANA in the patients with orbital disease was 17.8％(65/366),obviously lower than that in the controls(28.6％,76/266),the difference also being statistically significant(x2 =10.389,P=0.001).The positive rate of anti-ENA antibody in the patients with orbital disease was 4.6％(17/366),also obviously lower that that in the controls(9.0％,24/266),with statistically significant difference as well(x2 =4.866,P=0.027).No anti-dsDNA antibody was detected in the patients with orbital disease.Only three patients with orbital disease(0.8％,3/366)were found ANCA positive,and no statistically significant difference was found in comparison with the controls(3.0％,8/266; x2 =3.127,P=0.077).Conclusions Elevated IgG4 level was commonly seen in the patients with orbital disease,where as autoantibodies were negative in the most of the patients,indicating that IgG4 might correlate with orbital disease,and part of orbital disease may belong to the IgG4-related orbital disease.

The preparation of electrophoretic microcolumn and its application to the electrophoretic separation of amino acids with a 2-mm I.D. fused-silica microcolumn packed with uniform quartz microncrystal prepared by hydrothermal synthesis are reported. With 1.5 mmol/L disodium phosphate buffer solution (pH 11.5) containing 30% (V/V) methanol, the tryptophan, phenylalanine and tyrosine were separated by the microcolumn electrophoresis and detected by an UV spectrophotometer without derivatization. The limits of detection were 0.038, 0.21 and 0.20 mol/L, respectively. The separation efficiency of tryptophan was 4.4×104 plates/m. The sample capacity of the electrophoretic microcolumn achieved 35 μL. The precisions of the microcolumn electrophoresis were satisfactory. The thermal effects of the electrophoretic microcolumn that without packing, packed with 360 μm quartz sand and with 9 μm length quartz microncrystal were discussed, respectively. It was found that the electrophoretic microcolumn packed with quartz microncrystal was able to inhibit Joule heat, increase sample capacity and enhance detection sensitivity. The microcolumn electrophoresis is one of the high-performance separation techniques for an in-situ, real-time and portable electrokinetic flow analysis system.

Objective To investigate the mechanisms of phagocytosis of virulent Leptospira by peritoneal macrophages of guinea pigs,andevaluatetheroleof innateimmuneinthepathogenesisof leptospirosis. Methods Peritoneal macrophages of guinea pigs were extracted. Three specific inhibitors ( microfilament inhibitor cytochalasin D,microtube inhibitor colchicine and PI3K signalling pathway inhibitor LY294002) were added respectively to the macrophages 1 h before the infection of virulent Leptospira interrogans serovar Lai type strain Lai in vitro.Meanwhile, control group without inhibitor was established.Phagocytosis was observed by laser scanning confocal microscopy and phagocytic rates were evaluated by flow cytometry 3 h after infection.ResultsThe phagocytic rates of control group, cytochalasin D group, colchicine group and LY294002 group were (38.98 ± 0.91)%,(23. 99 ± 1. 40) % ,(40.81±0.91)% and (39.64 ±3.56) %, respectively.The phagocytic rate of cytochalasin D group was significantly lower than that of control group (P < 0. 05), while those of colchicine group and LY294002 group were not significantly different from that of control group (P >0.05). ConclusionMicrofilaments play an important role in the phagocytosis of strain Lai by peritoneal macrophages,but the process is independent on PI3K signalling pathway,and microtubes play little part during the phagocytosis.

Hemolysins of Leptospira interragans have been shown to be the virulence factor in the pathogenesis of leptospirosis and 10 potential hemolysin genes were charecterized by genomic annotation of L.interrogans serovar.Lai strain 56601. In the present study, the LA0202 gene supposed to encode one of the new potential hemolysin was cloned and the protein encoded was purified. The purified protein was shown to have highly hemolytic activity as demonstrated on the sheep blood agar plate. It was also confirmed that the LA0202 protein-mediated hemolysis on sheep erythrocytes was osmotically protected by PEG6000. Meanwhile, this protein could induce pore formation on sheep erythrocytes and cause damages on the membrane of human L-02 liver cells. In addition, it could induce apoptosis of human L-02 liver cells after treatment of cells with this protein for 24 hours. It is evident that LA0202 protein acting as a pore-formong hemolysin can induce cytotoxic damage on mammalian cells.

Objective · To isolate phages which can fight against extended-spectrum β-lactamase (ESBLs)-producing Escherichia coli (E. coli), and provide basic research for establishment of E. coli phage library and treatment of bacterial infection. Methods · Samples collected from sewage were co-cultured with 93 ESBLs-producing E. coli strains. A phage named JDEC001 was isolated by double agar overlay plaque assay. The biological characteristics, complete genome sequence and comparative genome analyses of JDEC001 were studied respectively. Results · JDEC001 belongs to the lytic phage as a member of the Caudovirales order, Podoviridae family. It has high activity at pH from 5 to 11 and with temperature from 0 to 39 ℃ .Whole-genome sequencing of JDEC001 demonstrated double-stranded DNA genome of 38745 bp with GC content of 49.93%, which encoded 46 open reading frames. The comparative genomics also showed that there was no virulent genes or antibiotic resistant genes in its genome. Conclusion · The phage JDEC001 against ESBLs-producing E. coli was isolated and purified, with good stability in a broad range of pH and temperature.

Objective:To investigate the relationship between the levels of vascular endothelial growth factor (VEGF) and homocysteine (Hcy) in serum and diabetic retinopathy (DR) and diabetic nephropathy (DN).Methods:A total of 211 patients with type 2 diabetes mellitus (T2DM) who were treated in Beijing Tongren Hospital from February to July 2019 were selected as the case group, including 72 patients with T2DM (T2DM group), 45 patients with DR (DR group), 49 patients with DN (DN group), 45 patients with DR and DN (DR+DN group); 76 healthy people were selected as the control group. The levels of serum VEGF and Hcy were measured in all subjects. The course of diabetes, body mass index, waist to hip ratio, fasting blood glucose, glycosylated hemoglobin (HbA 1c), triglyceride(TG), total cholesterol(TC), low density lipoprotein-cholesterol(LDL-C), high density lipoprotein-cholesterol(HDL-C), urinary microalbumin/creatinine (ACR), urinary immunoglobulin G/creatinine (IGU/CR), urinary transferrin/creatinine (TRU/CR), urinary α1-microglobulin/creatinine (α1/CR), serum urea nitrogen, serum creatinine, hypersensitive C-reactive protein (hCRP) and erythrocyte sedimentation rate (ESR) were also observed in the case group. Results:The VEGF level of the T2DM group, DR group, DN group and DR+DN group was 90.02(61.24, 118.52), 124.38(81.50, 170.28), 133.19(78.84, 168.49), 124.08(79.82, 187.33)ng/ml respectively, which were significantly higher than that of the control group 50.31(21.10,67.74)ng/ml(all P<0.05); Compared with the T2DM group, the VEGF level in the DR group, DN group and DR+DN group increased significantly (all P<0.05). The level of Hcy in the DN group and DR+DN group [(12.58±3.66), (11.91±2.42) μmol/L, respectively] was higher than that in the control group [(10.44±2.09) μ mol/L], and the difference was statistically significant ( P<0.05). There was no statistically significant difference in VEGF and Hcy levels in different stages of DR ( U=264.00, t=-0.43, P>0.05). The Hcy level of DN patients in the group of massive proteinuria was higher than that in the group of microalbuminuria [(15.00±1.87) vs (11.79±3.76) μmol/L, t=-2.82, P=0.01].VEGF was positively correlated with ACR, TRU/CR and IGU/CR ( r=0.23, 0.19, 0.17, all P<0.05),while Hcy was positively correlated with serum urea nitrogen, serum creatinine, ACR, TRU/CR, IGU/CR and α 1/CR ( r=0.35, 0.44, 0.22, 0.19, 0.21, 0.29, all P<0.05). Conclusions:The level of VEGF in the serum of DR and DN patients increased, suggesting that VEGF may play a role in the development of DR and DN, but there was no significant difference in the level of VEGF in patients with different stages of DR and different urinary albumin excretion rate of DN.The level of serum Hcy in DN patients increased, and that was higher in massive proteinuria group, suggesting that serum Hcy may have clinical significance in the diagnosis and monitoring of DN.

Objective:To investigate the serum metabolites and their metabolic characteristics of patients with optic neuritis.Methods:Case-control study. From January 2021 to January 2022, 9 serum specimens of diagnosed patients with optic neuritis were collected in Department of Neurology from Beijing Tongren Hospital and 9 healthy subjects as the control. Among them, there were 5 females and 4 males in the optic neuritis group, aged (35.8±12.9) years; there were 5 females and 4 males in the healthy control group, aged (32.6±8.6) years. Liquid chromatography-mass spectrometry was used to detect metabolites in serum of healthy control and patients with optic neuritis. The principal component analysis (PCA) and orthogonal partial least-squares discriminination analysis (OPLS-DA) were used to analyze the differential metabolites . The variable importance projection value of OPLS-DA model and the P value of t-test was applied to find the different metabolites. Results:Thirty-seven metabolites were finally identified from serum samples. Four metabolites with variable important in projection (VIP) values larger than 1 and P values less than 0.05 were teased out, three metabolites, LysoPC (P-16∶0), LysoPC (16∶0), LysoPC (P-18∶0) belonge to phospholipid and one metabolite was L-Threonine, they were all down-regulated. The area under curve were 0.951, 0.889, 0.963 and 0.944, respectively. Conclusion:Based on metabonomic analysis, some metabolites in serum have changed, which can provide basis for biomarkers screening of optic neuritis.

This paper has summarized the practical results of the mixed teaching model of Medical Microbiology in Shanghai Jiao Tong University School of Medicine. The model sets learning objectives under the concept of outcome-based education (OBE), and designs the curriculum under the guidance of objectives. In the reform practice of 120 students, the courses were implemented by using high-quality online resources to design four teaching links of "offline preview-online learning-offline participation-learning expansion" from three aspects of "pre-class-in-class-after-class", and the learning objectives were assessed by the teaching evaluation. Results showed that, compared with the traditional class, the performance of the students from reform class were significantly improved, the scores of more difficult questions were significantly higher than those of the traditional class, and the teaching quality was significantly improved. The mixed teaching model of Medical Microbiology has exploratory significance for solving the current situation of insufficient student participation and low learning efficiency in medical basic courses.