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Objective Graves' disease is the most common autoimmune thyroid disease and its prevalence and clinical manifestations are disparate between females and males. Costimulatory molecules play an essential role in regulating autoimmune responses. The objective of this study was to determine if expression of inhibitory molecules was correlated with treatment by dihydrotestosterone (DHT) in an BALB/c mouse model of experimental autoimmune Graves' disease.Methods Female BALB/c mice were immunized three times with thyroid stimulating hormone receptor A-subunit encoded by adenovirus to establish a Graves' disease model. Three different doses of DHT or a matching placebo were administered by implantation of slow-release pellets a week before the first immunization. Four weeks after the third immunization, the mice were euthanatized, and then the spleen and thymus were removed. Total thyroxine and free thyroxine levels in serum of mice were detected using a radioimmunoassay kit. Real-time polymerase chain reaction was performed to estimate the expression of costimulatory molecules in lymphocytes from the spleen and thymus. Flow cytometry was used to analyze the percentage of CD4 T cells in splenic lymphocytes. Quantitative data were compared with unpaired -tests. Correlation between two variables was analyzed using Analysis of Variance.Results Treatment with DHT can dramatically reduce total thyroxine and free thyroxine levels. Higher expression of programmed death-1 was found in the spleen of Graves' disease mice receiving 5 mg of DHT treatment (0.635±0.296 . 0.327±0.212; =2.714, =0.014), similarly, T-cell immunoglobulin domain and mucin domain 3 (TIM-3) in both the spleen (1.004±0.338 . 0.646±0.314; =2.205, =0.022) and the thymus (0.263±0.127 . 0.120±0.076; =3.221, =0.004) also increased after 5 mg of DHT treatment compared with the parallel placebo model mice. Moreover, the percentage of CD4 T cells declined in the splenic lymphocytes of Graves' disease mice treated with 5 mg of DHT (19.90%±3.985% . 24.05%±2.587%; =2.804, =0.012). A significant negative association was observed between expression of TIM-3 in the spleen and serum levels of total thyroxine (=-0.7106, =0.014) as well as free thyroxine (=-0.6542, =0.029).Conclusion This study demonstrates that DHT can ameliorate experimental autoimmune Graves' disease, which may occur by up-regulating expression of programmed death-1 and TIM-3 and inhibiting development of CD4 T cells.
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Objective To establish a real time PCR assay with a novel fluorescence quencher for identification of mutation of clarithromycin -resistance gene of Helicobacter pylori.Methods Two mutations of 23S rDNA gene in Helicobacter pylori,No.2142 and 2143,were chosen as targets for detection,and then the primers and the probe with a novel fluorescence quencher were designed.The genome DNA of Helicobacter pylori was extracted,and then detected by real time PCR reported here.Meanwhile,the specificity,reproducibility and sensitivity of the assay were evaluated.Finally,the real time PCR described here,the real time PCR based on TaqMan,and a sequencing assay were applied to detect 55 Helicobacter pylori strains isolated from clinical specimens,respectively.The results from three assays were compared with each other in order to further evaluate the applicability of this assay in clinic.Results It indicated that the mutation points related to clarithromycin -resistance,A2142G and A2143G,were identified by real time PCR with a novel fluorescence quencher rapidly and accurately.Moreover the coefficient of variation was less than 5%.The limit of detection was 100 copies/reaction.While this assay was applied directly to detect 55 Helicobacter pylori strains,the results were in accordance with those obtained from a TaqMan real time PCR and a sequencing assay,respectively.Conclusion The real time PCR described here was a simple,reliable and accurate approach and substituted for the TaqMan real time PCR for identification of two mutation points of clarithromycin -resistance,A2142G and A2143G in Helicobacter pylori.Thus,a novel tool for diagnosis of gene mutation was provided and the results might be regarded as a substantial evidence for clinical individual therapy.
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Rheum tanguticum from the same area was divided into 8 types of variation according to the plant morphology, content differences of free anthraquinones, combined anthraquinones, double anthrone were studied. The results showed that the functional components of different variation types were significantly different. The average content of free anthraquinone combined anthraquinone was 2.10-6.71 and 15.43-22.04 mg•g⁻¹, respectively. The average content of sennoside A plus sennoside B was 32.88-42.36 mg•g⁻¹. There were significant differences among the difference of 10 kinds of active components, except for sennoside B and physcion glycoside. Interred with the content and proportion of functional components, type B and type E might be potential special medicinal germplasm for diarrhea attack product, type G and type H might be a potential special medicinal germplasm for clearing heat and detoxifing, type C and type F might be potential special medicinal germplasm for activating blood circulation to dissipate blood stasis, type A and type D might be potential special medicinal germplasm with anastaltic funtion. The conclusion laid the foundation for the directional cultivation of fine varieties of special purpose of rhubarb.
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Objective To analyze the MP infection status in children with respiratory tract disease and the correlation with gen‐der ,age ,season and clinical conditions .Methods To investigate the clinical data retrospectively of 655 children with respiratory tract infection from January to December 2013 .Results The positive rate of MP antibody was 48 .09% with a higher incidence in girls than boys ,and those above 3 were more susceptible to it .Winter and spring were the peak seasons .The older group had a higher positive rate of MP antibody ,together with high morbidity of MPP and LP ,the differences were statistically significant (P<0 .05) ,while the younger group was inclined to get higher WBC count and percentage of increasing CRP and neutrophils ,the differ‐ences were statistically significant(P<0 .05) .Conclusion MP is a common pathogenic bacteria causes respiratory tract infection in children above 3 years ,especially in winter and spring .The antibody positive rate rise with age and the infected children are more likely to have pneumonia meanwhile the younger group has higher WBC count ,in which more cases get higher level of CRP and neutrophils .
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<p><b>OBJECTIVE</b>To study the electromyographic and genetic characteristics in children with Charcot-Marie-Tooth disease type 1 (CMT1).</p><p><b>METHODS</b>Routine electromyography and nerve conduction were performed in 24 children with CMT1. Polymerase chain reaction (PCR) combined with restriction enzyme digestion was used to detect gene duplication on chromosome 17p11.2-12. Ten healthy children served as the control group.</p><p><b>RESULTS</b>The peripheral nerve conduction velocity slowed or disappeared in all of the 24 patients (100%). The lesions of the sensory nerves were more severe than the motor nerves, and the lesions of the lower limbs were more severe than the upper limbs. Of 72 muscles detected, 40 (56%) showed neurogenic lesions. The older the patients, the more severe the muscle lesions. Specific junction fragments (1760 bp) were identified in 13 (54%) out of 24 patients, but were not identified in the healthy controls.</p><p><b>CONCLUSIONS</b>The electromyographic changes are characterized by peripheral nerve conduction velocities slowing and neurogenic lesions of muscles in children with CMT1. The PCR combined with restriction enzyme digestion may be a simple and accurate method for gene diagnosis of CMT1.</p>
Subject(s)
Adolescent , Child , Female , Humans , Male , Charcot-Marie-Tooth Disease , Genetics , Electromyography , Neural ConductionABSTRACT
<p><b>OBJECTIVE</b>To develop a small-caliber vascular grafts and study its morphologies, mechanical properties and biocompatibility.</p><p><b>METHODS</b>The effects of electrospinning conditions on the microstructure and porosity of the resulting scaffolds were investigated for preparation of a small-caliber (4 mm) polyurethane vascular grafts with optimum microstructures and mechanical properties. The mechanical properties and biocompatibility of the prepared grafts were evaluated.</p><p><b>RESULTS</b>The polyurethane vascular grafts showed a three-dimensional reticular structure consisting of nanofibers, with an average porosity of (51.48∓4.47)% and tensile strength of 5.85 ∓ 0.62 MPa. The grafts provided a better long-term support than e-PTFE graft for endothelial cell growth and endothelialization.</p><p><b>CONCLUSION</b>The polyurethane vascular prosthesis possessed favorable microstructures, excellent mechanical properties and good biocompatibility for potential clinical application.</p>
Subject(s)
Humans , Biocompatible Materials , Chemistry , Blood Vessel Prosthesis , Cell Adhesion , Materials Testing , Mechanical Phenomena , Polyurethanes , Chemistry , Porosity , Prosthesis Design , Tensile StrengthABSTRACT
Objective To develop a rapid, sensitive and specific assay method, based on multiplex real time PCR for identifying Vibrio cholerae and distinguishing Vibrio cholerae O139 serotype from Vibrio cholerae. Methods Cholera toxin A subunit gene (ctxA) and glycosyltransferase gene (LPSgt) were chosen as targets according to Vibrio cholerae and Vibrio cholerae O139 serotype,and then the primers and TaqMan-MGB probe were designed. The 5'end of probes was labeled with FAM and VIC fluoresceins respectively while the 3' end of probes was labeled with MGB. The PCR reaction was optimized systematically. Then the specificity, sensitivity and reproducibility of multiplex real time PCR were estimated. Finally, multiplex real time PCR was applied to detect the clinical specimens. Results Vibrio cholerae was identified by multiplex real time PCR accurately and quickly,which could distinguish Vibrio cholerae O139 serotype from Vibrio cholerae. Vibrio cholerae was identified positive for primer pairs and probes from ctxA gene, and Vibrio cholerae O139 serotype tested was positive for LPSgt gene. Meanwhile, none of other bacteria was identified. Sensitivity of the test was 2 × 102 cfu/ml. When this assay was applied directly to identify 45 clinical specimens, results showed that 10 were positive to Vibrio cholerae, in which 4 clinical specimens were positive to Vibrio cholerae O139 serotype. All the results were the same to the one that had been obtained from the conventional assays. Conclusion Our rsults showed that the multiplex real time PCR was a reliable,accurate and feasible method for identifying Vibrio cholerae that carrying toxin gene and distinguishing Vibrio cholerae O139 serotype from Vibrio cholerae. This method could be applied to the cholera surveillance, prevention and control system for identifying and distinguishing Vibrio cholerae in the labs.
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<p><b>BACKGROUND AND OBJECTIVE</b>Monitoring the therapeutic effects of radiotherapy for nasopharyngeal carcinoma (NPC) is critical to providing individualized treatment. This in-vivo study was initially designed to evaluate the therapeutic effect of radiotherapy using 18F-fluorodeoxyglucose positron emission tomography with computed tomography (18F-FDG PET-CT) imaging.</p><p><b>METHODS</b>18F-FDG PET-CT imaging was performed on all of the 10 nude mice bearing NPC xenografts before radiotherapy, and early-phase and delayed-phase PET-CT images were performed on 7 of the 10 mice. All mice were randomly divided into either a control group or a radiotherapy group. The 5 mice in the control group were immediately killed after the imaging and pathology were performed. After receiving radiotherapy of 12 Gy, 5 animals in the radiotherapy group were given 18F-FDG PET-CT imaging on days 2, 4, and 6, and then were killed for pathologic evaluation. Regions of interest (ROI) technology was used to measure the tumor target/non-target (T/NT) ratio and the volume of the tumors.</p><p><b>RESULTS</b>The average T/NT ratios of early- and delayed-phase imaging were 1.806 +/- 0.532 and 1.777 +/- 0.597, respectively, with no significance (P > 0.05). For the radiotherapy group, the average T/NT ratios for 18F-FDG PET-CT before radiotherapy, and on days 2, 4, and 6 after radiotherapy, were 1.735 +/- 0.466, 1.818 +/- 0.396, 1.096 +/- 0.101, and 0.604 +/- 0.108, respectively, The tumor volumes were (1.48 +/- 0.27) cm3, (1.57 +/- 0.31) cm3, (1.59 +/- 0.31) cm3 and (1.60 +/- 0.29) cm3, respectively. The average T/NT ratios of day 6 after radiotherapy and the other time points were significant (P < 0.05). The average death ratio of the tumor cells was (93.00 +/- 7.42)% after 6 days of post-radiotherapy.</p><p><b>CONCLUSIONS</b>18F-FDG PET-CT imaging can be used for the early assessment of radiotherapeutic effect of NPC in vivo. Day 6 after radiotherapy may be an appropriate time point for the imaging. However, the T/NT ratio measurement of delayed-phase imaging might make no sense for the diagnosis of NPC.</p>
Subject(s)
Animals , Humans , Male , Mice , Carcinoma, Squamous Cell , Diagnostic Imaging , Pathology , Radiotherapy , Cell Line, Tumor , Fluorodeoxyglucose F18 , Ki-67 Antigen , Metabolism , Mice, Inbred BALB C , Mice, Nude , Multimodal Imaging , Methods , Nasopharyngeal Neoplasms , Diagnostic Imaging , Pathology , Radiotherapy , Neoplasm Transplantation , Positron-Emission Tomography , Random Allocation , Tomography, X-Ray Computed , Tumor Burden , Radiation EffectsABSTRACT
<p><b>BACKGROUND</b>Children with anisometropic amblyopia are often noncompliant with traditional treatment including spectacules and contact lenses. This study was to evaluate the long-term efficacy of excimer laser in situ keratomileusis (LASIK) for children with high anisometropic amblyopia.</p><p><b>METHODS</b>A retrospective analysis of 24 children with high unilateral anisometropic amblyopia, who underwent LASIK during the period between August 2000 and September 2005 in our hospital, was conducted. The mean age of these children was (7.4 +/- 1.9) years (range 5 - 14 years) and the mean follow-up period was (33.3 +/- 14.2) months (range 18.5 - 74.2 months). After LASIK, visual acuity, refraction and far or near stereoacuity were analyzed. Near stereoacuity was measured by the random-dot butterfly stereogram and the pre-school random-dot stereogram, while far stereoacuity was measured by the synoptophore with Yan's random-dot stereogram.</p><p><b>RESULTS</b>Mean preoperative uncorrected visual acuity was 0.06 +/- 0.05, while mean postoperative uncorrected visual acuity was elevated to 0.43 +/- 0.33. Mean preoperative best-corrected visual acuity was 0.26 +/- 0.22, while mean postoperative best-corrected visual acuity was elevated to 0.67 +/- 0.40. For patients with myopic anisometropia, preoperative mean spherical equivalent refraction was (-8.01 +/- 2.70) D while postoperative value significantly reduced to (-1.32 +/- 2.47) D. For patients with hyperopic anisometropia, preoperative mean spherical equivalent refraction was (+7.35 +/- 1.55) D while postoperative value significantly reduced to (+3.30 +/- 0.86) D. These results demonstrated that there was statistical difference in these parameters between preoperative and postoperative tests. At the last follow-up, 20 patients had near stereoacuity, and the mean near stereoacuities measured by the random-dot butterfly stereogram and the preschool random-dot stereogram were (149.00 +/- 152.93)'' and (201.05 +/- 235.94)'', respectively. In contrast, 11 patients had far stereoacuity, and the mean far stereoacuity measured by the synoptophore was (210.91 +/- 217.51)''. Three cases of patients developed mild corneal complications.</p><p><b>CONCLUSION</b>LASIK for the correction of high anisometropia in children with monocular amblyopia may be safe and effective, and this can help effectively conduct postoperative amblyopia training and improve the cure rate.</p>
Subject(s)
Adolescent , Child , Child, Preschool , Female , Humans , Male , Amblyopia , General Surgery , Anisometropia , General Surgery , Keratomileusis, Laser In Situ , Methods , Retrospective Studies , Treatment Outcome , Visual AcuityABSTRACT
Objective The present study was conducted to investigate the infection of Lyme disease, Spotted fever, Ehrlichiosis (anaplasmosisin) in wild animals and ticks in the mountain areas of Zhejiang province. Methods Nested polymerase chain reaction was used to amplify specific DNA sequences of Lyme spirochetes, Spotted fever group rickettsiae, Ehrlichia (anaplasma) from samples of mice and ticks. Results 14 positive samples were identified from 121 mice and 105 groups of ticks. Among mice samples, one positive 5S-23S rDNA intergenic spacer of Borreia burgdorferi and two 5' fragments of Ehrlichia (anaplasma) 16S rDNA were obtained. 11 positive results were detected from tick samples including three 5S-23S rDNA intergenic spacer regions of Borreia burgdorferi and eight 5' fragments of Spotted fever group rickettsiae outer member protein A gene. One group of adult ticks, Haemaphysalis longicornis, which had been collected from eastern mountain area were detected to have co-infected with Lyme spirochetes and Spotted fever group rickettsiae. The positive sequences of 5S-23S rDNA intergenic spacer and ompA gene were tested and analyzed as Lyme spirochetes while rickettsia which was closely related to Borrelia valaisiana and R. massilliae. Conclusion This was the first report about co-infection of Lyme spirochetes and Spotted fever group rickettsiae found in the same group of adult Haemaphysalis longicornis. It is very important to strengthen the surveillance program on tick-borne infectious disease and their pathogenic in vectors, wild animals and targeted high risk groups and to differentiate the clinical manifestation and diagnosis to extend the knowledge of tick-borne infectious diseases in Zhejiang.
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<p><b>OBJECTIVE</b>To investigate the expression of RANTES in venous ulceration.</p><p><b>METHODS</b>From patients with lower limb venous ulceration, patients with non-ulcerous venous insufficiency, and healthy individuals, peripheral blood was collected from the lower limb veins for measurement of RANTES mRNA using RT-PCR.</p><p><b>RESULTS</b>In the ulceration group, the expression of RANTES mRNA was significantly increased as compared with the other two groups (P<0.01). RANTES mRNA expression was also significant higher in the non-ulcerous group than the control group (P<0.01).</p><p><b>CONCLUSION</b>High expression of RANTES mRNA may be one of the important mechanisms of venous ulceration.</p>
Subject(s)
Adult , Aged , Female , Humans , Male , Middle Aged , Young Adult , Chemokine CCL5 , Blood , Genetics , RNA, Messenger , Genetics , Reverse Transcriptase Polymerase Chain Reaction , Varicose Ulcer , Blood , Venous Insufficiency , BloodABSTRACT
<p><b>OBJECTIVE</b>To discuss the selection of the initial surgery extent for differentiated thyroid cancer (DTC) without metastasis.</p><p><b>METHODS</b>The clinical data of 504 cases with DTC, who accepted the surgical treatment from Jan 1995 to Dec 2004, were analyzed and studied. There were 329 cases without metastasis. The operative extents less than total thyroidectomy were performed on 93 cases (92.1%) with stage T(1), 166 cases (88.3%) with stage T(2), 22 cases (91.7%) with stage T(3) and 12 cases (75.0%) with stage T(4). The recurrence situation after the initial surgery was compared between different T-stage groups and between different surgical extents, total and less than total thyroidectomy.</p><p><b>RESULTS</b>The recurrence of DTC was found in 37 cases of the follow-up cases (8.9%), including 29 cases without metastasis in the initial surgery. There was no significant difference in the recurrent rate between T(1) and T(2) groups (P>0.05). The significant difference was found in recurrent rate between T(1) and T(3) or T(4) groups, T(2) and T(3) or T(4) groups (P<0.05). No significant difference in the ratio of the initial surgical extent less than total thyroidectomy was found between stage T(1) and T(2) cases without metastasis (P>0.05). The rate of the recurrent laryngeal nerve injury was 1.2%. The transient hypoparathyroidism happened in 2% of the cases, without the permanent hypoparathyroidism.</p><p><b>CONCLUSIONS</b>The surgical extent less than total thyroidectomy, especially subtotal thyroidectomy, is rational and available to stage T(1) and T(2) cases of DTC without metastasis. It can effectively remove the tumor and avoid postoperative complications. Total thyroidectomy should be performed on stage T(3) and T(4) cases of DTC.</p>
Subject(s)
Adolescent , Adult , Aged , Aged, 80 and over , Child , Female , Humans , Male , Middle Aged , Follow-Up Studies , Neoplasm Metastasis , Neoplasm Recurrence, Local , Retrospective Studies , Thyroid Neoplasms , Pathology , General Surgery , Thyroidectomy , Methods , Treatment OutcomeABSTRACT
<p><b>OBJECTIVE</b>To examine vibrio cholera (V.C) in aquatic products of littoral area, Zhejiang Province and to provide scientific evidence for administration of aquatic products and cholera epidemic control.</p><p><b>METHODS</b>All 990 samples of aquatic products collected from local markets, eateries and aquafarms in three chosen areas. Samples were proliferated in alkaline liquid medium, and purified in NO: 4 medium, the isolations were identified biochemically, and phenotype of strains were defined by phagocyte and coagulation with V.C. diagnostic serum. Three virulence genes (ctx, ace, zct) of the isolated strains were detected by polymerase chain reaction (PCR).</p><p><b>RESULTS</b>There were 1.41% samples caught by V.C., having a carrying rate highest in turtles of 8.9%. 14 strains were defined as three serogroups, and the numbers of Inaba, Ogawa, and Hikojima types were 2, 2, 10 respectively. Virulence genes had detected in 9 of 12 stains. All genes were detected in 5 strains, only ZOT genes in 3 strains, and both CTX and ACE genes in 1 strain.</p><p><b>CONCLUSIONS</b>Aquatic products from inshore in Zhejiang Province caught with V.C. strains might be divided into three serogroups. Most of them should be virulence genes. Cholera epidemic outbreak might be caused by those contaminated products.</p>