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Objective To explore the feasibility of artificial silastic framework(SF)in repair of large area of tracheal wall defects.Methods Twenty healthy adult dogs with tracheal defects for 2.5 cm?6.0 cm-3.0 cm?6.0cm were randomly and equally divided into experimental group(repaired with SF combined with sternohyoid fasciae)and control group(repaired with T-silastie tubule combined with sternohyoid fascial flap).After the operation,the animals were sacrificed at the 4th,8th,16th,24th, and 48th weeks respectively for harvesting the tracheae that were used for tracbeoscopically observing in- flammatory reaction of the repaired defect area and light microscopically observing epithelium healing on the repaired defect area.Results In the experiment group,the repaired trachea was smooth,without proliferation of granulation;and at the 8th week,the repaired defect area was covered with epithelial cells,with good functional recovery of respiration,phonation and deglutition.In the control group,there was obvious proliferation of granulation on the tracheal surface near anterior and posterior ends of T-silas tic tubule.The animals were under asphyxia to die with extraction of T-silastic tubule.Conclusions SF has excellent tracheal skeletal function.In the meantime,SF combined with sternohyoid fasciae is a simple but effective method for repair of large area of tracheal wall defects.

To explore the expression potential of heterogeneous genes using the backbone of infectious bronchitis virus (IBV) Beaudette strain, the ectodomain region of the Spike gene (1,302 bp) of IBV H120 strain was amplified by RT-PCR and replaced into the corresponding location of the IBV Beaudette strain full-length cDNA. This recombinant was designated as BeauR-H120(S1). BeauR-H120(S1) was directly used as the DNA template for the transcription of viral genomic RNA in vitro. Then, the transcription product was transfected into Vero cells by electroporation. At 48 h post-transfection, the transfected Vero cells were harvested, and passaging continued. A syncytium was not observed until the recombinant virus had passed through four passages. The presence of rBeau-H120(S1) was verified by the detection of the replaced ectodomain region of the H120 Spike gene using RT-PCR. Western blot analysis of rBeau-H120 (S1)-infected Vero cell lysates demonstrated that the nucleocapsid (N) protein was expressed, which implied that rBeau-H120(S1) could propagate in Vero cells. The TCIDs0 and EIDs0 data demonstrated that the titer levels of rBeau-H120(S1) reached 10(590+/-0.22)TCID50/mL and 10(6.13+/-0.23)EID50/mL in Vero cells and 9-day-old SPF chicken embryos, respectively. Protection studies showed that the percentage of antibody-positive chickens, which were vaccinated with rBeau-H120(S1) at 7 days after hatching, rose to 90% at 21 days post-inoculation. Inoculation provided an 85% rate of immune protection against a challenge of the virulent IBV M41 strain (103EID50/chicken). This recombinant virus constructed using reverse genetic techniques could be further developed as a novel genetic engineering vaccine against infectious bronchitis.
Animals ; Cercopithecus aethiops ; Chick Embryo ; Chickens ; Coronavirus Infections ; veterinary ; virology ; Infectious bronchitis virus ; chemistry ; genetics ; growth & development ; metabolism ; Poultry Diseases ; virology ; Protein Structure, Tertiary ; Spike Glycoprotein, Coronavirus ; chemistry ; genetics ; metabolism ; Transfection ; Vero Cells

Objective To explore the value of 11C-choline PET/CT in patients with hepatic spaceoccupying lesions that have an indeterminate diagnosis by 18F-fluorodeoxyglucose (FDG) PET/CT. Methods A total of 25 liver masses in 20 patients with an indeterminate diagnosis based on 18F-FDG PET/CT were enrolled. Regional 11C-choline PET/CT scan was performed in all of the patients. Lesions with intense 11C-choline uptake were considered as positive. The semiquantitative maximum standardized uptake value(SUVmax) was measured and the tumor-to-liver (T/L) radioactivity ratio was calculated. The Mann-Whitney test,Kruskal-Wallis test and crosstabs x2-test were performed by using SPSS version 11.5. Results Of the 25 lesions,21 were proven to be hepatocellular carcinomas (HCC),3 hemangiomas,and 1 parasitic granuloma. The sensitivity of 11C-choline PET/CT for the detection of HCC was 66.7% (14/21). 11C-choline PET/CT had a higher sensitivity for well differentiated HCC than moderately and poorly differentiated HCC on a patient basis (8/9 vs 2/5,respectively). There were significant differences of 11C-choline T/L ratios between the HCC positive group,HCC negative group and benign lesion group ( 1.70 ± 0.35,0.86 ± 0.15,and 0.36 ± 0.18,x2 = 19.00,P ＜0.01 ). The lesion size and alphafetoprotein (AFP) level between the HCC positive and negative groups had no significant difference respectively ( Mann Whitney U = 39.00,P ＞0.05,and U=16.00,P＞0.05,respectively). Conclusions 11C-choline is complementary to 18F-FDG PET/CT for the detection of HCC,especially for well differentiated HCC.

Objectives To investigate the clinical features,prognosis and risk factors of patients with cryptococcal meningitis.Methods Totally 146 patients with cryptococcal meningitis who were hospitalized in Huashan Hospital from January 2000 to December 2006 were enrolled in this study.The clinical data including diagnosis and misdiagnosis,experimental and etiology tests,treat- ments and prognosis from all the patients were analyzed retrospectively.Results Among the 146 patients enrolled in the study,78 patients(53.4%)had concomitant diseases.The misdiagnosis rate of all patients was 72.6%(106/146).The positive rate of cerebrospinal fluid(CSF)India ink smear was 59.6%(87/106),while 43.2%(63/146)cases of cryptococcus neoformans culture in CSF was positive.The positive rate of Latex agglutination test(LAT)was 91.7%(134/146)in CSF among all patients.The treatments were as follows:combination of Amphotericin B(AmpB)or its lipid formula- tions with flucytosine(5-FC)(98 cases),including combination with Fluconazole initally(62 cases), single therapy of Fluconazole(13 cases).Ommaya implanted for lateral cerebral ventricle drainage(53 cases)and AmpB intrathecal injection(53 cases).The average dose of AmpB is 3.06 g.The course of treatment lasted from 12 weeks to 20 months.There were 104 patients(71.2%)cured,27(18.5%) improved,15(10.3%)died and 34(23.3%)relapsed.Conclusions High misdiagnosis rate is common in patients with cryptococcal meningitis.Immunodeficiency is the major risk factor for cryp- tococcal meningitis.CSF LAT is the most sensitive diagnostic test.Early diagnosis,combination of AmpB with 5-FC antifungal therapy and control of acute intracranial hypertension are the keys to im- prove prognosis of cryptococcal meningitis.

Objective To investigate the expression of Toll-like receptor 3 (TLR3) on dendritic cells(DCs) derived from peripheral blood mononuclear cells(PBMCs) of chronic hepatitis B(CHB) patients and to explore the mechanism of sustained infection of hepatitis B virus (HBV). Methods Twenty CHB patients were randomly screened in the study,and ten healthy persons were recruited as controls.The monocytes isolated from peripheral blood of candidates were incubated with recombinant human granuloeyte macrophage colony-stimulating factor (rhGM-CSF) and rhIL-4 to induce the DCs generation and proliferation.Then the phenotype of DCs was identified by micro- scope.The expressions of the phenotypes[histocompatibility leukocyte antigen(HLA)-DR,CD80, CD86,CD83]of immature and mature DCs were measured by flow cytometer.Furthermore,the ex- pression of TLR3 on mature DCs(mDC) and immature DCs(imDC) was determined by flow cytometry and Western blot analysis respectively.Results As for healthy volunteers,the expressions of CD80, CD86,HLA-DR and CD83 on DCs at the 7th day,which were(82.35?8.67)%,(79.61?10.08)%, (92.79?8.48)% and (83.76?5.47)% respectively,were significantly higher than those at the 5th day which were(28.31?8.79) %,(31.17?11.23)%,(27.61?10.28)% and (23.46?11.53)% respec- tively(P0.05).The expression of TLR3 on imDC was significantly higher than that on mDC at control group (P0.05).And the expression of TLR3 on imDC in CHB patients group was significantly lower than that of control group(P

OBJECTIVETo screen the stage-specific expression proteins during rats spermatogenesis, and to investigate the beta-actin expression and localization in the tissues of rat testicular.

METHODSHighly enriched type A spermatogonia, pachytene spermatocytes and round spermatids were isolated by STAPUT method (sedimentation velocity at unit gravity, with 2% - 4% BSA gradient in DMEM/F12 medium) respectively to get the total proteins. The difference of protein expression between the three kinds of cells was analyzed by two-dimensional electrophoresis. Then the distribution of beta-actin in rat testicular tissues was investigated using specific anti-beta-actin antibodies by immunohistochemical method.

RESULTSbeta-actin was identified as a stage-specific expression protein by two-dimensional electrophoresis. beta-actin protein was more strongly expressed in type A spermatogonia and pachytene spermatocytes, but not in round spermatids. The immunohistochemical results showed that beta-actin was mainly located in the cytoplasm of type A spermatogonia and pachytene spermatocytes and in the nuclei of nearly mature spermatids.

CONCLUSIONbeta-actin protein is a stage-specific expressed protein and may play an important role in spermatogenesis.

Actins ; biosynthesis ; Animals ; Electrophoresis, Gel, Two-Dimensional ; Male ; Mass Spectrometry ; Rats ; Rats, Sprague-Dawley ; Spermatogenesis ; physiology ; Testis ; cytology ; metabolism

The infection status of anisakid larvae was examined in 290 marine fish of 25 species and in 108 cephalopods of 3 species purchased in Bayuquan region, Yingko city nearby the coast of the Bohai Sea from may to August 1992. A total of 7,327 larvae were collected from 156 fish of 19 species and 8 squids of one species. The 3rd-stage larvae of Anisakis simplex were collected from 121 fish (63.4%) of 15 species (N = 191) and from 8 squids (14.8%) of one species (N = 54), and they were total, 5,992 (81.8%). Out of remaining 1,335 larvae, 154 (2.1%) were classified as Thynnascaris type B from 23 fish of 4 species, 1,013 (13.8%) as Thynnascaris type C from 79 fish of 13 species. 164 (2.2%) as Hysterothylacium China type V from 20 fish of 4 species, 3 (0.04%) as Raphidascaris from 3 fish of 2 species and one was Pseudoterranova decipiens larva.
Animal ; Anisakiasis/veterinary* ; Anisakiasis/parasitology ; Anisakiasis/epidemiology ; Anisakis/isolation & purification ; Anisakis/classification* ; China ; Fish Diseases/parasitology* ; Fish Diseases/epidemiology ; Fishes ; Larva ; Seawater ; Squid/parasitology*

The chitosan/PHEA-blended hydrogels were prepared from PHEA and chitosan in various blend ratios. The water contents of the hydrogels were in the range of 50%-80% (wt). The attachment and growth of fibroblast cells(L929) on the hydrogels were studied. The results indicated the PHEA content in hydrogels has great effect on cell attachment but has little effect on the growth of L929 cells.
Animals ; Biocompatible Materials ; chemistry ; Cell Adhesion ; physiology ; Cell Division ; physiology ; Cells, Cultured ; Chitin ; analogs & derivatives ; chemistry ; Chitosan ; Fibroblasts ; cytology ; physiology ; Hydrogel, Polyethylene Glycol Dimethacrylate ; chemistry ; Mice ; Peptides ; chemistry ; Water ; chemistry

OBJECTIVEImpulse noise was adopted in adult rats to built acute deafferent animal model. Differential proteomics techniques were applied to detect the changes of protein expression in the auditory cortex before and after the noise exposure.

METHODSThirty adult SD rats were divided into three groups: normal group, rats with acute noise exposure and rats 28 days recovery after noise exposure (n=10/group). All animals were exposed to impulse noise at 156 dB for 50 pulses with a rise-time of 100 µs and duration of around 0.25 ms. ABR was used to evaluate the auditory function. The two-dimensional gel electrophoresis (2-DE) and matrix-assisted laser desorption ionization time-of-flight mass spectrometer (MALDI-TOF-MS) were used to identified the differential protein expression.

RESULTSCompared with the normal group, ABR thresholds were found significantly increased at 2, 4, 8, 16, 32 kHz (P<0.05) in the acute and recovery groups. There was a 40-60 dBSPL ABR threshold shift at all tested frequencies immediately after impulse noise exposure. There was a partial recovery of ABR thresholds at 7 day to 28 days after impulse noise exposure. In addition, it seemed that the thresholds were rather stable and no further ABR threshold recovery was observed from 14 day to 28 days after the impulse noise exposure. Using differential proteomic techniques, 36 spots containing 27 proteins were revealed and identified in auditory cortex. Those proteins are related to cytoskeleton, neurotransmission, energy supply, mitochondrial function and synaptic remolding.

CONCLUSIONSImpulse noise may influence the function of microtubule transport and cell metabolism, there after affect the neurotransmission of auditory neurons. The compensatory changes such as pre- and postsynaptic or such related functional changes may also happen in auditory cortex after the deafferentation treatment.

Animals ; Auditory Cortex ; metabolism ; physiopathology ; Evoked Potentials, Auditory, Brain Stem ; Hearing Loss, Noise-Induced ; metabolism ; physiopathology ; Male ; Proteomics ; Rats ; Rats, Sprague-Dawley