Objective To investigate the relationship between optic d isc hemorrhage and localized retinal never fiber layer defects (RNFLDs) in norma l-tension glaucoma. Methods In 83 patients with normal-tensi on glaucoma, the cumulative frequency and quadrantal distribution of optic disc hemorrhages were retrospectively analyzed. The neighboring relation between opti c disc hemorrhages and RNFLDs in a same quadrant and the changes of correspondin g retinal never fiber layer (RNFL) after the occurrence of optic disc hemorrhage s were observed by tridimensional photochromy of ocular fundus. Results (1) The occurrences and distribution of optic disc hemorrhages: 29 of 83(34.94%) patients (33 eyes) had totally 58 occurrences, including 39 in infer iotemporal area, 14 in superiotemporal area, and 5 in other area. (2) The relati onship of neighborhood between optic disc hemorrhages and RNFLDs: in the availab le tridimensional photochrome, 23 occurrences in 15 patients (16 eyes) were foun d with cuneiform RNFLDs in the same quadrant, in which 22 was near the border of cuneiform RNFLDs. (3) The changes of corresponding retinal never fiber layer (R NFL) after the occurrence of optic disc hemorrhages: the photochromes of 24 occu rrences in 20 patients (21 eyes) were kept well in the initial and the 2-year f ollow-up periods, while the changes of RNFL were found in each region correspon ding to the 19 occurrences (in inferiotemporal or superiotemporal area) in the i nitial photochrome, including 7 cuneiform defects with various sizes, and 12 dev eloped localized RNFLDs next to the initial hemorrhages in the optic disc. No ob vious localized RNFL corresponding to the other 5 occurrences (1 in inferiotempo ral, 1 in superiotemporal, and 3 in other areas) were found in the follow-up pe riod. Conclusion Optic disc hemorrhages in normal-tension gla ucoma occur mostly in inferiotemporal area, and secondly in superiotemporal area of optic disc, and the appearance of optic disc hemorrhages may suggest that th e localized RNFLDs would develop in the associated regions.

Objective To establish a specific,sensitive and applied method for the detection and differentiation of dengue virus types Ⅰ-Ⅳ,Japanese encephalitis virus and yellow fever virus.Methods Based on the genomes sequence analysis,6 pairs of primers were designed.The special capture probes of dengue virus types Ⅰ-Ⅳ,Japanese encephalitis virus and yellow fever virus were amplified,cloned and sequenced.Then the microwell plates were precoated using these capture probes,and the forward primers were labeled using biotin.The samples were then amplified using the biotin labeled forward primers and reward primers.The microwell plate hybridization was processed for detecting and differentiating the virus.The precoated DNA concentration,precoated time,hybridization temperature and hybridization time were optimized carefully.Results The A value of positive samples were over 0.5,while the average A value of the negative samples was less than 0.1.The S/N value exceeded 10.0.Sensitivity experiment suggested the method of PCR-ELISA could detect the virus RNA in 107 times dilution,while RT-PCR could detect the virus RNA in only 106 times dilution.The stability experiment of PCR-ELISA using DVⅠ suggested that the within-batch coefficient of variation was 6.21%,the between-batch coefficient of variation was 9.92%;the within-batch coefficient of variation in negative control was 1.92%,and the between-batch coefficient of variation in negative control was 3.68%.No visible changes were found on the performance of the coated microwell plates when stored in 4℃for 6 monthes.Conclusion PCR-ELISA is a more sensitive and specific method than RT-PCR is in the early detection and type identification of dengue Ⅰ-Ⅳ types virus,Japanese encephalitis virus and yellow fever virus.

Objective To screen and identify the dengue virus-specific antigens,then establish the ELISA detection method for dengue virus antibody.Methods Using bioinformatic software DNAStar and ANTHEPROT to analyze the hydrophilicity,flexibility,surface probability and antigenicity of dengue virus type 1-4,Japanese encephalitis virus and Yellow fever virus M,E and NSI protein amino acid sequence and also consider the influence of secondary structure.Then in accordance with epitopes localion and amino acid sequence similarity,forecast the share and specific epitopes.Reference the sequence information of different dengue virus strains in GenBank to analyze the epitopes conservative.Based on the results of bioinformatic analysis,5 specific epitopes were amplified and inserted into prokaryotic expression vector pMAL-C2x or pET32a.Then the vectors was transferred into E.coli Rosetta( DE3 ).lsopropyl-β-D-thiogalactoside(IPTG) was used to induce the expression of gene segments.SDS-PAGE were used to identify the expression proteins,and the antigenicity were tested using Western blot.Using the antigen selected by Western blot,ELISA method for dengue virus antibody detection was established.Results Eighty shared epitopes and 25 specific epitopes were forecasted,and 5 antigenic fragments encloude analyzed epitopes from dengue virus type 2 and 3 were expressed in E.coli successfully.One dengue virus type 1-4 shared antigens (Den-Ag5),one dengue virus type 2 and 4 shared antigens( DenAg3),one dengue virus type 1-3 shared antigens(Den-Ag2) and two dengue virus type 1,2 and 4 shared antigens( Den-Ag1,Den-Ag4)were conformed using Western blot.Using antigens Den-Ag5,Den-Ag1 and DenAg2,the ELISA method for dengue virus antibody detection were established.Conclusion Based on the bioinformatic analysis and Western blot verification,5 dengue virus specific antigen were conformed,and the ELISA detection method for dengue virus antibody were established.

PURPOSE: To produce animal models of Acanthamoeba keratitis and to evaluate the advantages and adaptation range of each of the three methods employed. MATERIALS AND METHODS: Mice and Wistar rats in three groups of 15 rats and 15 mice each were used to establish the models. Right corneas in group A were scratched and challenged with Acanthamoeba. Those in group B were scratched and covered with contact lenses incubated with Acanthamoeba. Those in group C received an intrastromal injection of Acanthamoeba. Five rats and 5 mice in each group were used for histopathological investigations and the other 10 in each group were used for clinical evaluation. The models were evaluated by slit lamp examination, microscopic examination and culture of corneal scrapings, HE staining of corneal sections, and pathological scoring of the infections. RESULTS: Four rats and 6 mice in group A, 7 rats and 8 mice in group B, and 10 rats and 10 mice in group C developed typical Acanthamoeba keratitis. CONCLUSION: Corneal scratching alone has the lowest infection rate, while scratching and then covering with contaminated contact lenses has a moderate rate of infection and most closely mimics what happens in most human infections. Intrastromal injection of Acanthamoeba gives a much higher infection rate and more severe Acanthamoeba keratitis.
Acanthamoeba/growth & development ; Acanthamoeba Keratitis/*parasitology/pathology ; Animals ; Contact Lenses/adverse effects/parasitology ; Cornea/parasitology/pathology ; Disease Models, Animal ; Female ; Male ; Mice ; Microscopy ; Rats ; Rats, Wistar

OBJECTIVETo investigate the effect of polysaccharide sulfate 916 (PS916) on the production of nitric oxide (NO) in ECV304 cells induced by tumor necrosis factor-alpha (TNF alpha), interleukin-1 beta (IL-1 beta) and H(2)O(2) in vitro.

METHODSProduction of NO in ECV304 cells was measured by the Griess method and the proliferation of cells was tested by the MTT method. The activity of NO synthase was detected spectrophotometrically.

RESULTSProduction of NO in ECV304 cells decreased after treatment with 40 ng/ml IL-1 beta and 40 ng/ml TNF alpha, but increased in the presence of H(2)O(2) 0.1 mmol/L. PS916 significantly enhanced NO production in ECV304 cells in a dose-dependent manner in the TNF alpha and IL-1 beta treated groups and decreased it in the H(2)O(2) treated group. Proliferation of ECV304 cells was inhibited by TNFalpha and H(2)O(2) and no effect was found in the IL-1 beta treated group. PS916 increased the proliferation of cells treated with TNFalpha and H(2)O(2) dose-dependently. In vitro, PS916 has no effect on the activity of NO synthase.

CONCLUSIONPS916 has a protective effect on ECV304 cells exposed to IL-1 beta, TNF alpha and H(2)O(2).

Cell Division ; drug effects ; Cell Line ; Cytokines ; pharmacology ; Dose-Response Relationship, Drug ; Endothelium, Vascular ; cytology ; drug effects ; metabolism ; Humans ; Hydrogen Peroxide ; pharmacology ; Interleukin-1 ; pharmacology ; Nitric Oxide ; biosynthesis ; Nitric Oxide Synthase ; drug effects ; metabolism ; Polysaccharides ; metabolism ; pharmacology ; Sulfuric Acids ; metabolism ; Tumor Necrosis Factor-alpha ; pharmacology

OBJECTIVETo study the effect of polysaccharide sulfate 916 (PS916) on neutrophil-endothelial cell adhesion.

METHODSCell adhesion was evaluated by testing neutrophil myeloperoxidase activity. Expression of adhesion molecule in human umbilical vein endothelial cell (HUVEC) was measured by ELISA. The neutrophil activation rate induced by N-formyl-methionyl-leucyl-phenylalanine (fMLP) was tested by nitroblue tetrazolium (NBT) reduction.

RESULTSTumor necrosis factor alpha (TNFalpha, 50 - 800 U/ml) increased the adherence of neutrophil to TNFalpha-stimulated HUVEC in a concentration and time dependent manner. PS916 (0.01 - 1.0 mg/ml) dose-dependently inhibited the adherence of neutrophils to TNFalpha-stimulated HUVEC. fMLP increased the activation rate of neutrophils independent of concentration. PS916 also inhibited the adherence of fMLP-activated neutrophils to HUVEC. Moreover, PS916 inhibited adhesion molecule expression in TNFalpha-stimulated HUVEC.

CONCLUSIONSPS916 inhibited neutrophil-endothelial adhesion. The mechanism of its action was partially related to suppressing the expressions of intercellular adhesion molecule-1 (ICAM-1) and vascular adhesion molecule-1 (VCAM-1).

Animals ; Cell Adhesion ; drug effects ; Cells, Cultured ; Endothelium, Vascular ; cytology ; Humans ; Intercellular Adhesion Molecule-1 ; analysis ; N-Formylmethionine Leucyl-Phenylalanine ; pharmacology ; Neutrophils ; drug effects ; physiology ; Polysaccharides ; pharmacology ; Rats ; Rats, Wistar ; Sulfuric Acids ; pharmacology ; Tumor Necrosis Factor-alpha ; pharmacology ; Vascular Cell Adhesion Molecule-1 ; analysis

Objective To observe the effects of excessive fluoride on osteocalcin and glucose metabolism in mice.Methods Thirty-two male C57 mice (body mass:18-24 g) were selected as study subjects which were randomized into four groups (8 mice in each group) according to their body mass by random number table method:0 mg/L group (control group),50 mg/L fluorine group,100 mg/L fluorine group and 150 mg/L fluorine group.Sodium fluoride in distilled water was freely taken by these animals to replicate fluorosis animal model.After 12 weeks,total osteocalcin,uncarboxylated osteocalcin,insulin and glucagon were measured by enzyme-linked immunosorbent assay,fasting blood glucose and glycated hemoglobin were measured by chemiluminescence.Results After 12 weeks of intervention with sodium fluoride,serum total osteocalcin,uncarboxylated osteocalcin,fasting blood glucose,glycated hemoglobin,insulin,and glucagon were significantly different between different groups (F =17.23,22.29,4.43,45.57,4.45,55.21,P ＜ 0.05).Total osteocalcin,uncarboxylated osteocalcin,fasting blood glucose,glycated hemoglobin,and insulin in the 100,150 mg/L fluorine groups were higher than those of control group [(30.02 ± 5.35),(35.22 ± 4.98) vs (20.23 ± 3.22) μg/L;(8.72 ± 1.34),(11.01 ± 1.02) vs (5.80 ± 1.60) μg/L;(7.53 ± 2.29),(8.53 ± 2.81) vs (4.99 ± 1.60) mmol/L;(6.74 ± 0.68),(7.12 ± 0.25) vs (4.95 ± 0.28) mmol/L;(2.65 ± 0.25),(2.74 ± 0.47) vs (2.13 ± 0.28) mU/L,P＜ 0.05].The serum glucagon levels in the 50,100,150 mg/L fluorine groups were lower than that in the control group [(20.90 ± 3.00),(23.68 ± 2.58),(21.63 ± 2.42) vs (38.61 ± 3.73) ng/L,P ＜ 0.05].Conclusion Excessive fluoride can lead to elevated osteocalcin level and abnormal glucose metabolism in mice.

Objective: To evaluate the effectiveness of different laboratory methods for the supplementary diagnosis of pulmonary tuberculosis（PTB）and provide reference data for the early diagnosis of PTB. Methods: A total of 298 suspected PTB patients, who were diagnosed and treated in the outpatient department of Shanghai Tongren Hospital from January 2016 to December 2018, were divided into 3 groups: active PTB (138 cases)，inactive PTB (43 cases) and non-PTB (117 cases) group. Sputum acid-fast staining, MGIT liquid culture system and Xpert MTB/RIF test were performed to detect the sputum specimens. The sensitivity and specificity were compared by Chi-square test. Results: The three methods showed certain significance for distinguishing active PTB, inactive PTB combined with non-PTB (χ 2 values were 89.08, 138.94 and 137.12 respectively, all P＜0.01). There was no significant difference for the positive rate of the three methods between inactive PTB and non-PTB. The sensitivities of acid-fast staining, MGIT liquid culture, Xpert MTB/RIF test and the combination of three methods in the diagnosis of active PTB were 45.7% (63/138), 63.8% (88/138), 65.4% (87/133) and 78.2% (104/133) respectively. The sensitivities of MGIT culture and Xpert MTB/RIF test were significantly higher than that of acid-fast staining (χ 2 was 35.79 and 11.26 respectively，all P＜0.01). There was no significant difference for the sensitivities between MGIT liquid culture and Xpert MTB/RIF test（χ 2 was 29.87, P＞0.05) . The sensitivity of combined detection was higher than that of single detection（χ 2 was 30.84, 64.62, 70.14, respectively, all P＜0.01）. The diagnostic specificities of the three methods and their combination were 99.1%（116/117), 98.3%（115/116）, 99.1%（113/114) and 97.3%（110/113）respectively. There was no significant difference for the specificities of the three methods. Conclusion High sensitivities of MGIT liquid culture and Xpert MTB/RIF test were shown in PTB diagnosis. Combined detection of the three methods may improve the sensitivity of detection.

BACKGROUND: Lung cancer is the cancer with the highest mortality at home and abroad at present. The detection of lung nodules is a key step to reducing the mortality of lung cancer. Artificial intelligence-assisted diagnosis system presents as the state of the art in the area of nodule detection, differentiation between benign and malignant and diagnosis of invasive subtypes, however, a validation with clinical data is necessary for further application. Therefore, the aim of this study is to evaluate the effectiveness of artificial intelligence-assisted diagnosis system in predicting the invasive subtypes of early‑stage lung adenocarcinoma appearing as pulmonary nodules. METHODS: Clinical data of 223 patients with early-stage lung adenocarcinoma appearing as pulmonary nodules admitted to the Lanzhou University Second Hospital from January 1st, 2016 to December 31th, 2021 were retrospectively analyzed, which were divided into invasive adenocarcinoma group (n=170) and non-invasive adenocarcinoma group (n=53), and the non-invasive adenocarcinoma group was subdivided into minimally invasive adenocarcinoma group (n=31) and preinvasive lesions group (n=22). The malignant probability and imaging characteristics of each group were compared to analyze their predictive ability for the invasive subtypes of early-stage lung adenocarcinoma. The concordance between qualitative diagnostic results of artificial intelligence-assisted diagnosis of the invasive subtypes of early-stage lung adenocarcinoma and postoperative pathology was then analyzed. RESULTS: In different invasive subtypes of early-stage lung adenocarcinoma, the mean CT value of pulmonary nodules (P<0.001), diameter (P<0.001), volume (P<0.001), malignant probability (P<0.001), pleural retraction sign (P<0.001), lobulation (P<0.001), spiculation (P<0.001) were significantly different. At the same time, it was also found that with the increased invasiveness of different invasive subtypes of early-stage lung adenocarcinoma, the proportion of dominant signs of each group gradually increased. On the issue of binary classification, the sensitivity, specificity, and area under the curve (AUC) values of the artificial intelligence-assisted diagnosis system for the qualitative diagnosis of invasive subtypes of early-stage lung adenocarcinoma were 81.76%, 92.45% and 0.871 respectively. On the issue of three classification, the accuracy, recall rate, F1 score, and AUC values of the artificial intelligence-assisted diagnosis system for the qualitative diagnosis of invasive subtypes of early-stage lung adenocarcinoma were 83.86%, 85.03%, 76.46% and 0.879 respectively. CONCLUSIONS Artificial intelligence-assisted diagnosis system could predict the invasive subtypes of early‑stage lung adenocarcinoma appearing as pulmonary nodules, and has a certain predictive value. With the optimization of algorithms and the improvement of data, it may provide guidance for individualized treatment of patients.
Adenocarcinoma/pathology* ; Adenocarcinoma of Lung/pathology* ; Artificial Intelligence ; Humans ; Lung Neoplasms/pathology* ; Multiple Pulmonary Nodules ; Neoplasm Invasiveness ; Retrospective Studies

The sternum is the pivotal component of the thoracic cavity. It is connected with the clavicle and ribs on the upper part and both sides respectively, and plays an important role in protecting the stability of the chest wall. Sternal resection usually results in a large segmental chest wall defect that causes the chest wall to float and requires sternal reconstruction. This paper reports a 62 years male patient with thymic squamous cell carcinoma with sternal metastasis, who underwent thymotomy, sternal tumor resection and autologous lilum graft combined with sternal reconstruction by titanium plate after relevant examination was completed and surgical contraindications were eliminated. The patient was followed up for 6 months, the respiratory and motor functions were normal and the thoracic appearance was good.