Objective To investigate the inhibitory effect of favipiravir (T-705) on canine distemper virus (CDV) replication in Vero cells and DH82 cells.Methods The growth curves of CDV-11 strains from canine and CDV-3 strains from mink in Vero cells and DH82 cells were determined with indirect immunofluorescence assay and 50％ endpoint titration.The viability of Vero and DH82 cells was determined using the Cell Counting Kit-8.CDV inhibition at different concentrations of T-705 at different time points was measured .Results Cytotoxicity data showed that there was a moderate decline of viability in Vero cells after T-705 treatment, but no apparent cytotoxicity in DH 82 cells.T-705 significantly inhibited the replication of CDV-3 and CDV-11 in both Vero cells and DH82 cells in the test range of 2.441-1250 μg/ml. T-705 exhibited effective and stable antiviral activity when given at different time points post virus challenge .Conclusion Our results demonstrate that T-705 has effective antiviral activity and may be a promising anti-CDV drug candidate .

A microfluidic chip with micropillar arrays for three-dimensional (3D) cell culture was designed and validated.The chip consisted of a polydimethylsiloxane (PDMS) channel plate and a glass cover plate.One cell culture chamber composed of two rows of micropillar arrays and two lateral channels for transporting the culture medium were integrated on the PDMS channel plate.The spacing between micropillars directly affects the chip performance, which is critical for the design of the chip.In this work, the spacing between micropillars was optimized by numerical simulation and experimental validation.With the optimized microfluidic chip, the mixture of cells and extracellular matrix mimics could be steadily injected into the cell culture chamber, the nutrients in the culture medium from the lateral channels could quickly diffuse into the chamber, and the cell metabolites could also timely diffuse out of the chamber.To test the stability of the microenvironment in the microfluidic chip, neural stem cells were three-dimensionally cultured.

Objective To investigate the sonographic features of follicular variant of papillary thyroid carcinoma (FV-PTC) and to decrease misdiagnosis rate. Methods Thirty-one patients with 35 FV-PTCs and 66 patients with 75 conventional PTCs (C-PTCs) were enrolled in this study. The sonographic features were reviewed retrospectively between the two groups with universally accepted standards. Results The sonographic features of 35 FV-PTCs included irregular shapes (6/35), anteroposterior to transverse diameter ratio A/T > 1 (7/35), spiculated margins (25/35), marked hypoechogenicity (0/35), hypoechogenicity (18/35), isoechogenicity (16/35), no calcification (15/35), microcalcifications (11/35), macrocalcification (9/35), color Doppler lfow patternⅠ(20/35), color Doppler lfow patternⅡ(10/35), color Doppler lfow patternⅢ(5/35). Irregular shapes, A/T>1, spiculated margins, marked hypoechogenicity, microcalciifcations, and color type Ⅱ were rarer in FV-PTCs than in C-PTCs, while isoechogenicity, no calciifcation, macrocalciifcation, and color type Ⅲwere more frequent in FV-PTCs than in C-PTCs. The differences of the above features were statistically significant [χ2=4.276, P=0.039; χ2=8.125, P=0.004; P=0.009 (Fisher′ s exact test); χ2=8.548, P=0.003;χ2=4.898, P=0.027,χ2=7.796, P=0.005;χ2=5.462, P=0.019;P=0.001 (Fisher′s exact test)] . During the preoperative ultrasonography, 20 of 35 FV-PTCs were diagnosed as malignancy, and others were misdiagnosed as benign nodules (misdiagnosis rate was 43%). The lymphatic metastasis rate of FV-PTCs was 29%(9/31), significantly lower than C-PTCs [62%(41/66),χ2=9.246, P=0.002]. In terms of the sonographic features of metastatic lymph nodes, there was no marked difference between FV-PTCs and C-PTCs. Conclusions Some FV-PTCs are lack of malignant features, and tend to be misdiagnosed frequently when coexisting with benign thyroid nodules. Observing the echogenicity, color lfow characteristics and other features of each thyroid nodule and cervical lymph node with multiple views may decrease the misdiagnosis rate.

OBJECTIVETo investigate the clinicopathologic features, immunophenotyping, differential diagnoses and prognosis of histiocytic sarcoma (HS).

METHODSThe clinical and pathologic findings of 4 cases of HS were reviewed. The samples were used for paraffin section, HE stain, immunohistochemistry stain by EnVision method, electron microscope observation. Follow-up information was available in all patients.

RESULTSThe age of patients, 2 males and 2 females, ranged from 22 to 65 years old (median, 43.25 years). The sites of involvement included lymph node (2 cases), skin or soft tissue (1 case) and colon (1 case). The tumor cells were widespread infiltration, diffused distribution, no adhesion to each other. Tumor cells were middling and large, round, orbicular-ovate, polygon, epithelium appearance, plentiful cytoplasm and acidophilia, cystose. Nucelus was round, orbicular-ovate, dissymmetry. Nuclear chromatin was vacuole appearance, basophilia nucleolus, caryocinesia and pathological mitotic figure. Three of the cases showed conjugate nuclei, increased pleomorphism with multinucleated tumor giant cell formation. Focal cytoplasmic with foamy appearance was identified in 2 cases. One case demonstrated foci of spindly sarcomatoid appearance. Hemophagocytosis was identified in 2 cases. The tumor cells of 4 cases were often accompanied by various numbers of inflammatory cells. Immunohistochemical study showed that all cases were diffusely positive for α-1-ACT, CD68, CDl63 and lysozyme. Three of 4 cases also expressed CD45, CD45RO. The electron microscope results of 4 cases showed that the tumor cells were plentiful cytoplasm and a few cytolysosome in the cytoplasm, and no birbeck cytorrhyctes, cell-cell junction and digitation. Amongst the 4 patients with follow-up information available, three died of the disease 6-13 months after diagnosis. One patient, whose lesion was localized at the skin and soft tissue, survived at the present time.

CONCLUSIONHS was a scarce malignant tumor with mature histiocyte morphology and immunophenotype character. The diagnosis should be based on tissue morphology, immunohistochemistry and electron microscope observation to exclude other disorders.

Adult ; Aged ; Diagnosis, Differential ; Female ; Histiocytes ; pathology ; Histiocytic Sarcoma ; diagnosis ; pathology ; Humans ; Immunohistochemistry ; Immunophenotyping ; Male ; Microscopy, Electron ; Middle Aged ; Young Adult

Objective To retrospectively analyse the clinical data of off-pump coronary artery bypass grafting surgery(OPCAB)in our department and summarize the clinical experience of peri-operative management.Methods From January 2007 to May 2008,253 patients underwent OPCAB.Seventy-six cases with left main disease(including 18 cases of single left main lesion and 58 cases of complex lesion);13 cases had single vessel disease;32 cases had double vessels disease and 190 cases had triple vessels disease.Eighteen cases received emergency OPCAB.All vessel anastomosis were performed with coronary artery stablizer and shunt through median sternotomy.Resuits Three cases died after operation(1.2%).Intra-aortic balloon pump was used in 11 cases.Mechanical ventilation time was 3-168 h[(24.1±22.3)h],and ICU stay time was 1-14 day[(2.8±2.0)d].Postoperative hospital stay time was 8-42 days[(15.6±6.3)d].Conclusion OPCAB is reliable for most bypass cases.With appropilate peri-operative management,a satisfied result will be acceptable.

Objective To summarize the clinical experiences of 1386 patients undergoing off-pump coronary artery bypass grafting(OPCAB)during Jan 2002 to Mar 2008.Methods OPCAB was done through midsternumtomy under ordinary-temperature anesthesia after radial artery,left breast artery and vena saphena magna were taken.Patients over 70 years old accounted for 32.35%,and the oldest was 99 years old.Emergency and subemergency OPCAB was done in 111 cases,re-do CABG with off-pump technique was done in 52 patients.Results 24 cases died during perioperation,with mortality of 1.73%and all other patients discharged from the hospital.Following operation,the symptoms of angina pectoris were relieved and the life quality was much beaer than that before operation.Conclusion OPCAB is feasible for older patients,who are complicated with other chronic diseases for elder patients.Skillful technique of surgeon,right indications for OPCAB and management of peri-and postoperation are key to success of operation.

Objective To evaluate the feasibility and clinical value of combined clonidine and sleep-deprivation induced seizures for ictal brain SPECT imaging in patients with epilepsy. Methods Fiftytwo epilepsy patients were given oral clonidine plus sleep-deprivation to induce seizures with video-electroencephalogram (VEEG) monitoring. Forty-seven patients were selected as control group, whose seizures were induced by sleep-deprivation only. 99Tcm-ethylcysteinate dimer (ECD) was injected within 30 s since a clinical sign and/or a typical EEG discharge of epilepsy was recognized. Brain SPECT was performed 30 min after 99TcmECD injection. X2-test was performed by using software SPSS 10. 0. Results One to two hrs after oral intake of clonidine plus sleep-deprivation, 75% (39/52) patients were induced seizures, including 92.3% (36/39) with subclinical seizures and 7.7% (3/39) with clinical seizures. Ictal brain SPECT localized the lesions with high uptake of 99Tcm-ECD in 37 (94.9%) patients. In control group, 38.3% ( 18/47) were induced epileptic seizures, including 77.8% (14/18) with subclinical seizures and 22.2% (4/18) with clinical seizures. The induction rate of epileptic seizures in clonidine plus sleep-deprivation group was significantly higher than that of control group (X2 = 13.614, P ＜ 0.01 ). However, there was no significant difference in clinical seizures between the two groups (X2 = 1.253, P ＞ 0.05 ). Conclusions The combination of oral intake of clonidine and sleep-deprivation could increase the induction rate of epileptic seizures and it is effective for epilepsy SPECT imaging.

To investigate the changes in neurological symptoms and signs, as well as serum copper, serum ceruloplasmin after hepatic transplantation in patients with Wilson's disease, neurological symptoms and signs, serum copper, serum ceruloplasmin before and after hepatic transplantation in 18 patients with Wilson's disease were observed, and those changes were followed up in 20 non-operative controls treated with penicillamine. Our results showed that the neurological symptoms and signs, serum copper and serum ceruloplasmin were improved in the operative group but deteriorated in the non-operative control group. Our study showed that hepatic transplantation is better than penicillamine in the treatment of Wilson's disease.
Adolescent ; Adult ; Ceruloplasmin ; metabolism ; Child ; Copper ; blood ; Female ; Follow-Up Studies ; Hepatolenticular Degeneration ; blood ; surgery ; Humans ; Liver Transplantation ; Male ; Treatment Outcome

Background Haze formation is a key factor of vision reduce following corneal refractive surgery.Transforming growth factor-β1 (TGF-β1) and α-smooth muscle actin (α-SMA) are documented to participate in haze formation.Laboratory study showed that bevacizumab can not only inhibit corneal neovascularization,but also promote the healing of corneal epithelial basement membrane.However,the impact of bevacizumab on corneal healing after Off-flap epipolis laser in situkeratomileusis (Off-flap Epi-LASIK) is unclear.Objective The present study was to investigate the inhibition effect of bevacizumab on corneal haze after off-flap Epi-LASIK and its active mechanism.Methods Off-flap Epi-LASIK was performed in 24 adult pigmented rabbits and these rabbits were randomized into three groups.Bevacizumab of 0.1 ml (2.5 mg) was subconjunctivally injected 10 minutes after surgery in 16 rabbits and the same amount of bevacizumab was repeatedly injected 4 days after the initial injection in 8 eyes of 16 eyes.In addition,equivalent amount of normal saline solution was used in the same way in the other 8 rabbits.Another 2 health rabbits were used as the blank controls.Operative eyes were examined by slit lamp biomicroscope daily after surgery and haze was scored based on SundarRayde criteria.Corneas were obtained 4 weeks after operation for hematoxylin & eosin and periodic acid Schiff staining.Expressions of TGF-β1 and α-SMA in corneal tissue were detected by immunochemistry.Results Corneal epithelium healed completely in all eyes 4-5 days after operation.The haze scores were lower in the bevacizumab single injection group and repeat injection group than those in the normal saline solution group (P＜0.05) in 1 week and 4 weeks after operation.However,no significant difference was seen in the haze scores between the bevacizumab single injection group and repeat injection group (P ＞ 0.05).The hostopathological examination showed that the fibrosis response of cornea tissue was slight in the bevacizumab single injection group and repeat injection group comparison with the normal saline solution group.At 1 week after operation,the expression levels of TGF-β1 were (49.8 ± 2.1) PU and (38.6 ±4.4) PU in the bevacizumab single injection group and repeat injection group,and those of 4 weeks were (37.7 ±4.8) PU and (28.3 ± 3.5) PU,indicating significant decrease in the TGF-β1 expression compared with (65.1 ±5.3) PU and (51.6±2.2) PU of the normal saline solution group in both 1 week and 4 weeks (P＜0.01).The expression levels of α-SMA in corneas were (67.2±10.0) PU and (32.7±3.1) PU at 1 week,and (34.2±5.7) PU and (22.8±3.0) PU at4 weeks after operation in the bevacizumab single injection group and repeat injection group,which were significantly lower than (87.8±7.7) PU and (59.4±5.6) PU in the normal saline solution group in both 1 week and 4 weeks (P＜0.05).Meanwhile,the expression levels of TGF-β1 and α-SMA were declined in the bevacizumab repeat injection group compared with single injection group (P＜0.01).Periodic acid Schiff staining exhibited that the basement membrane of cornea was intact and continued in bevacizumab injection group.But corneal basement membrane was discontinuous in the normal saline solution group.Conclusions Subconjunctival injection of bevacizumab downregulates the expressions of TGF-β1and α-SMA in cornea after Off-flap Epi-LASIK and thus prevents haze formation.

Genes related to trehalose biosynthesis from a bacterial strain Micrococcus luteus which can convert partially hydrolyzed starch into trehalose were cloned.Full sequence of gene (MtreY) encoding trehalose maltooligosyl trehalose synthase (MTSase) and partial sequence of gene (MtreZ) encoding maltooligosyl trehalose trehalohydrolase (MTHase) were got using PCR combined non-random shotgun method.Sequence analysis of MtreY predicts a 2370bp open reading frame encoding a protein of 790 amino acids with a predicted molecular weight of 86734 Da.Homologous analysis shows that this new gene has the same conservative motifs with ?-amylase family enzymes.The MtreY gene was expressed in E.coli, and the expression product has the anticipative enzyme activity.