AIM: To analyze the pathogens and drug sensitivity of chronic dacryocystitis in order to provide evidence for clinical drug use.
METHODS:Lacrimal secretion of 171 cases with chronic dacryocystitis was sampled for pathogenic bacteria culture identification and drug sensitivity test. Based on the results, the isolation rate of pathogens strains, the pathogens kind of chronic dacryoeystitis, main pathogens of chronic dacryocystitis, and sensitive drug for pathogens were analyzed.
RESULTS: The isolation rate of pathogens strains was 76. 61% ( 131 cases ). The pathogens constituting the chronic dacryocystitis were predominantly gram-positive coccus,the percentage was 72. 52% (95 cases), among which staphylococcus hominis occupied 27. 48% ( 36 cases), staphylococcus epidermidis 16. 79% (22 cases), streptococcus viridans 12. 98% (17 cases). The majority of these bacteria were sensitive to cefoperazone-sulbactam, tobramycin, gentamicin and levofloxacin. For gram -positive coccus, cefoperazone - sulbactam, gentamicin and tobramycin were the most sensitive drug. For gram-negative bacilli, cefoperazone - sulbactam, tobramycin and levofloxacin were most sensitive drug.
CONCLUSION: Staphylococcus hominis is the main pathogen of chronic dacryocystitis, tobramycin can be used as the first choice for local treatment of chronic dacryocystitis.

Adult ; Arsenic Poisoning ; etiology ; Arsenicals ; Humans ; Male ; Skin ; injuries ; Skin Absorption ; Wounds and Injuries

Objective To analyze the dynamic evolution of brain MRI in patients with mitochondrial myopathy,encephalopathy,lactic acidosis,and stroke-like episodes (MELAS) syndrome.Methods A retrospective study was performed on 58 MELAS cases with pathologically and (or) molecularly confirmed diagnosis.MRI were repeated within 60 days after the onset of stroke-like episodes (SLE) and the evolution changes of cerebral lesions were accessed.Brain atrophy index (BAI) was calculated in the remission stage from 31 patients with MELAS,and the correlation between BAI,age and disease duration was analyzed.Results The proportion of lesions expansion,migration and shrink within 30 days after the onset of SLE was 64.1％ (25/39),10.2％ (4/39),17.9％ (7/39),respectively,and 13％ (3/23),21.7％ (5/23),56.5％ (13/23),between 30-60 days after the onset of SLE respectively.In the recovery stage of SLE,the BAI in 31 patients with MELAS was 15.2％ ±2.8％.The correlation coefficient between BAI and the age,total disease course and duration of encephalopathy was 0.329 (P =0.043),0.405 (P =0.012) and 0.649 (P =0.000).Conclusions Brain atrophy in the studied MELAS patients gradually develops and strokelike lesions shrink with progression of the disease.However,the migration of lesions is persistent.

Background Research showed that transforming growth factor-β2 (TGF-β2) promotes scar formation.But its mechanism in scarring after glaucoma filtration surgery is worthy of studying.Objective This study was to investigate the effect of TGF-β2 on myofibroblast transition of human Tenon fibroblasts (HTFs) and scarring after glaucoma filtration surgery.Methods Tenon capsular tissue was obtained from 3 patients with strabismus during the surgery and was incubated in DMEM with 10％ fetal bovine serum (FBS).The cells were collected and passaged in the free-serum medium for 24 hours,and then 1,2,5,10,20 μg/L TGF-β2 was added into the medium respectively,to induce the transformation of HTFs,and 2 μg/L or 5 μg/L TGF-β2 was used to treat the HTFs for 6,24,48 and 72 hours.The control group was not treated with TGF-β2.The expressions of α-smooth muscle actin (α-SMA) and phosphorylation of the signaling proteins (pSmad2) in HTFs were detected by Western blot assay.The expressions of α-SMA and F-actin were located by cell immunofluorescine technique under the confocal immunofluorescence microscopy.Cell contractility was determined by collagen gel contraction assays.This study was approved by Ethic Committee of Institute of Surgery Research of Daping Hospital,and informed consent was obtained from each patient or custodian initial of the study.Results The expression of α-SMA protein in the HTFs was increased significantly after the treatment of TGF-β2 in comparison with the control group and reached a peak at 24-48 hours.The α-SMA expression was gradually weakened in the 10 μg/L TGF-β2 groups.Little of α-SMA and F-actin were expressed in the control group.However,strong staining for α-SMA and F-actin were observed in the 1,2 and 5 μg/L TGF-β2 groups and then the staining weakened at the concentration of 10 μg/L.In addition,pSmad2 showed a stronger expression in the 2 μg/L TGF-β2 group than that in the PBS group and FBS group,with the strongest expression in 30 minutes through 2 hours.The untreated gel contracted (78.00±3.13)％ from its initial size,and contraction in the 1,2,5,10 μg/L TGF-β2 group were (63.88±1.78)％,(20.69±0.65)％,(19.49-±0.54)％,(16.24±0.84) ％,respectively,TGF-β2 increased HTFs contraction significantly (Fgroup =859.400,P =0.000).Conclusions TGF-β2 can induce transdifferentiation of Tenon fibroblast into myofibroblast and increase cell contractility,with a concentration-dependent and time-dependent pattern to an extent.It may be the mechanism of scar formation after glaucoma filter surgery.

Background Our previous study demonstrated that the aptamer S58 specifically targeted transforming growth factor-β receptor Ⅱ (TβRⅡ) and inhibited the transdifferentiation of human Tenon capsule fibroblasts (HTFs) mediated by transforming growth factor-β (TGF-β).Chitosan-nanoparticles (CS-NP) are good drug carriers,but the efficacy and safety of CS-NP/aptamer complexes deserve attention.Objective The aim of this study was to synthesize a novel CS-NP/aptamer complex called CS (S58)-NP and investigate its properties and applicability.Methods Human Tenon capsule tissue was obtained from patients during strabismus surgery,and HTFs were cultured and passaged using the explant culture method.The fourth to tenth generations of cells were used in the experiment.Different concentrations of CS-NP were used to prepare the CS(S58)-NP by the ionic cross-linking method with a surface charge rate (N/P) for S58 of 10,20,30 or 40.The particle size and Zeta potential were measured by the Zeta analyzer.The shape and distribution of CS (S58)-NP particles were examined under the scanning electron microscope.The binding of CS-NP with S58 and resistance of CS (S58)-NP to DNase Ⅰ were examined by agarose gel eletrophoresis.The release rate of S58 from CS (S58)-NP in PBS was quantitatively analyzed by a ultraviolet spectrophotometer.The cytotoxicity of CS(S58)-NP to HTFs was evaluated by detecting the production of lactate dehydrogenase (LDH).Results The Zeta analyzer showed that the particle size of CS (S58)-NP was 130-270 nm and its electric potential ranged from + 16 to +28 mV.The CS (S58)-NP particles appeared spherical with an even distribution under the scanning electron microscope.The mean encapsulation efficiency of CS(S58)-NP was 88.9％,89.3％,91.7％ or 90.5％,respectively,when the N/P was 10,20,30 or 40.After being encapsuled by CS-NP,S58 could resist the degradation from DNase I.Its total releasing level in PBS increased with the lapse of time,with a maximum releasing speed at 24 to 36 hours.The total releasing level reached 100％ at 96 hours.With increaseing concentrations of CS(S58)-NP,the relative releasing level of LDH in HTFs suspension gradually elevated with a significant difference among the groups (F =588.018,P =0.000),with the highest released LDH level at 50 nmol/L of CS(S58)-NP (12.853％ ±0.375％).Conclusions CS-NP provides a protective and slow-releasing effect on the S58 aptamer.CS (S58)-NP shows a good biocompatibility with HTFs with a low cytotoxicity at a concentration of ＜50 nmol/L.CS(S58)-NP could be used to inhibit TGF-β induced transdifferentiation of HTFs in the future.

Objective The comparison of the clinical role of stress-redistribution/reinjection with dual isotopes of 99Tcm-methoxyisobutylisonitrile (MIBI) and 201TI in the detection of myocardial viability.Methods One hundred and sixty patients with clinically suspicious coronary artery disease (CAD) were included.All had intravenous injection with 740 MBq of 99Tcm-MIBI.Pharmacological challenge with dobu-macological challenge with dobutamin,111 MBq of 201Tl Was injected to all.Myocardial SPECT images were performed in all at 10-min (stress) and 3-h (redistribution/rest) after injection.The 201Tl(37 MBq)would be given to those patients with myocardial perfusion defect at stress images by 201Tl and were demon-strated by both 201Tl(redistribution) and 99Tcm-MIBI (rest).Coronary angiography (CAG) Was performed within two weeks.X2-test was used with SAS 6.12.Results Coronary artery abnormalities were found in all with 76 patients had one vessel disease,51 had two and 33 had three.Of the 152 patients who had myo- cardial perfusion defect during stress images,63 had redistribution by both 201TI and 99Tcm-MIBI.5 had re-distribution by 201Tl only.9 had redistribution by 99Tcm-MIBI only,and 75 had no redistribution in 201Tl or 99Tcm-MIBI images.The sensitivity of detection myocardial viability with myocardial SPECT images between 201Tl at redistribution (66.0%,68/103) and 99Tcm-MIBI at rest (69.9%,72/103) were insignificant (x2=O.36.P>0.05).Of the 75 patients who did not have redistribution in 201Tl or 99Tcm-MIBI images.34.7% (26/75)had myocardial perfusion when reinjection of 201Tl.In all,there were eight false negative myocardial perfusion SEPCT images.Three were triple vessel disease,one Was two, three were one, and the other was patent collateral circulation.Conclusions Stress.redistributed/reinjection 201TI myocardial perfusion SPECT imaging is superior to stress 201Tl/rest 99Tcm-MIBI simultaneous dual-isotopic myocardial imaging in the detec-tion of myocanrdial viability.

Objective To investigate the expression of lipid rafts (LRs) and actin cytoskeleton (F-actin) in the peripheral blood B lymphocytes of patients with systemic lupus erythematosus (SLE).Methods Peripheral blood mononuclear cells (PBMCs) were separated by Ficoll-Hypaque.B lymphocytes were isolated by positive selection from PBMCs.Membrane staining for LRs was achieved with FITC-conjugated cholera toxin B (CTB).The level and distribution of LRs were studied by flow cytometry and confocal microscopy.Staining for F-actin was carried out with Rhodamine phalloidin.The expression of F-actin was analyzed by confocal microscopy.In an in vitro examination,the effect of Leflunomide on lipid rafts in B lymphocytes from SLE was analyzed.Disease carried out was measured using the SLE disease activity index (SLEDAI).Analysis of the enumerical data was performed using ANOVA or paired-samples t test.Correlation was examined by Pearson's rank correlation test.Results The number of CTB-binding lipid rafts in B cell from active SLE patients or from SLE patients in disease remission.who were treated with immunosuppressive drugs was higher than B cells from healthy controls [(59+4)％,(51±5)％,(33±4)％,F=9.21,P=0.001].Confocal microscopy revealed that in B cell from healthy controls,lipid raft was found to be small and uniformly distributed on the plasma membrane.F-actin was found mainly in the cortical region of the cells.This pattern was different from the pattern seen in B cells from patients with SLE,which presented with stronger staining and irregular large clustering of LRs,with a decrease in F-actin levels.In addition,the number of CTB-binding LRs in B cells from the active SLE patients was correlated significantly with the SLEDAI score (r=0.632,P=0.028).Furthermore,thein vitro results showed that leflunomide treatment reduced the number of CTB-binding LRs in B cell from SLE patients [(48±5)％ vs (39±5)％,t=2.29,P=0.048].Conclusion The altered expression of Lipid raft and F-actin can been seen in B lymphocytes in SLE,and modulation of LRs and F-actin expression may be a potential approach in the treatment of SLE.

Objective To observe the protection of human keratinocyte cell line, HaCaT cell, from UVA damage by resveratrol and its possible mechanism. Methods HaCaT cells were incubated with or without 0.01 mmol/L or 0.1 mmol/L resveratrol after exposure to 5 J/cm2 UVA irradiation. Unirradiated HaCaT cells-without the treatment with resveratrol served as the control. After another 24-hour culture, MTT assay was used to detect the proliferation of cells, RT-PCR and Western-blot to measure the iNOS mRNA and protein expression respectively, electron microscopic technique to observe the changes in cell ultrastructure. Results After irradiation with UVA of 5 J/cm2, the proliferation of HaCaT cells decreased with the absorbance at 490 nm descending from 0.889±0.083 to 0.542±0.004, while a significant increase was observed in the relative expression level of iNOS mRNA and protein in HaCaT cells (1.532±0.041 vs 0.009±0.003, 1.331 ±0.046 vs 0.003±0.001, both P < 0.05) with the presence of typical apoptotic cells. The treatment with 0.01 and 0.1 mmol/L resveratrol significantly promoted the proliferation of irradiated cells with the absorbance at 490 nm being 0.753±0.435 and 0.892±0.173 respectively, but inhibited the mRNA (0.853±0.038 vs 1.532±0.041, 0.392±0.033 vs 1.532±0.041, both P< 0.05) and protein expression level (0.809±0.018 vs 1.331±0.046, 0.412±0.026 vs 1.331±0.046, both P< 0.05) of iNOS in irradiated cells, and the resveratrol of 0.1 mmol/L was more effective than that ofO.01 mmol/L in all tested parameters (P< 0.05). Furthermore, no apoptofic cells or necrotic cells were observed in irradiated ceils incubated with resveratrol. Conclusion Resveratrol effectively protects HaCaT cells from UVA damage, which may be related to the inhibition of UVA-induced iNOS expression by resveratrol.

BACKGROUND: Classic isolation method of bone marrow mesenchymal stem cells (BMSCs) is Percoll density gradient centrifugation. Blood cell component was removed. However, this method is complicated. Preparation density was needed when isolating dog bone marrow. Moreover, centrifugation was frequent, which had a great damage to cells. OBJECTIVE: To establish methods of the isolation, proliferation, culture and osteoinduction of canine BMSCs, and observe the in vitro proliferation and ability to osteoinduction differentiation. METHODS: 10 mL bone marrow was extracted from dog posterior superior iliac spine, heparin anticoagulation, diluted using Hanks juice, treated with 1.077 g/mL Ficoll solution 3 mL, and centrifuged at 2 000 r/min for 20 minutes. Karyocytes were absorbed to form white cloudlike layering interface, and then centrifuged twice using DMEM supplemented with fetal bovine serum, incubated at 12×10~4/cm~2 at 37 ℃ in a 5% CO_2 incubator. Following subculture, cells were incubated in DMEM containing dexamethasone, β-sodium phosphoglycerol and ascorbic acid 2-phosphate. Immunocytochemical staining and immunofluorescence staining were utilized to detect osteocalcin, osteopontin and type Ⅰ collagen expression in osteoblasts. Alkaline phosphatase staining and alizarin red staining were performed. RESULTS AND CONCLUSION: 1.077 g/mL Ficoll density gradient centrifugation was used to isolate karyocytes that were significant compared with Percoll solution. Obtained BMSCs had high purity, good growth and the mean doubling time was 24 hours. Following in vitro osteogenic incubation of dog BMSCs, osteocalcin, osteopontin and type Ⅰ collagen showed positive expression. Alkaline phosphatase staining demonstrated bluish-green cytoplasm. Alizarin red staining showed red nodes in extracellular matrix, and could differentiate into osteoblasts in vitro.

AIM: To explore the expression of cytokeratin 20 (CK-20)target gene in peripheral blood of patients with colorectal cancer. METHODS: Total cellular RNA was extracted from whole blood of cancer patients and control groups, including 10 healthy adults, 17 benign gastrointestinal disease patients, 32 cases of colorectal cancer. The CK-20 gene expression was measured by reverse transcription PCR. RESULTS: 1. CK-20 mRNA was not detected in benign gastrointestinal disease patients and healthy adults. 2. In the colorectal carcinoma group, CK-20 mRNA expression was negative in 2 case of patients at Dukes A. The CK-20 mRNA positive expression can be detected in 4 patients among 15 cases at Dukes B, 6 out of 10 cases at Dukes C, 4 out of 5 cases at Dukes D. 3. The total positive percentage of CK-20 mRNA is 43 8% in 32 colorectal cancer patients; The positive percentage of CK-20 mRNA in patients at Dukes A and B is 23 5% and 6 7% in Dukes C, D patients CONCLUSION: It is a specific, sensitive and no damage method to detect the CK-20 target gene in patients with colorectal cancer. It is helpful for early diagnosis of tumor micrometastasis and for direction of chemotherapy.