The NSP2 gene of porcine reproductive and respiratory syndrome virus (PRRSV)S1 strain was partly amplified and cloned into a prokaryotic expression vector pGEX-6P-1 and a fusion protein GST-tNSP2 with molecular weight of 50 kDa was expressed in E.coli. The purified GST-tNSP2 protein showed a strong reaction with the PRRSV-positive sera in Western blot assay. Balb/c mice were immunized with the purified protein, and the splenocytes of the immunized mice were fused with murine myeloma cells SP2/0. After subcloning by 3 times, two hybridoma clones which produced McAbs steadily were screened by ELISA, named 3H3 and 2B5. They all reacted strongly with the PRRSV S1 infected Marc-145 cells in IFA, but not with the PRRSV SY0608 strain. Both of the McAbs belong to IgG1 isotype, and their light chains belong ? type. The expressed GST-tNSP2 protein and McAbs could be used for identification of PRRSV isolates and functional analysis of NSP2.

Objective:To express the EMCV 3AB gene by prokaryotic expression systerm,and prepare monoclonal antibodies against it. Method: NSP 3AB gene of Encephalomyocarditis virus (EMCV) was amplified and cloned into a prokaryotic expression vector pGEX-6P-1 and a recombinant protein 3AB with high antigenicity was expressed in E.coli. Balb / c mice were immunized by purified recombinant 3AB protein of inclusion-body, and the splenocytes of the immunized mice were fused with murine myeloma cells to produce hybridoma cell line. Results: After subcloning by 3 times, one strain of hybridoma cell line steadily secreting antibodies of 3AB protein was obtained, named 2D12. The McAb belongs to IgG1/?. The McAb and was confirmed by indirect immunofluorescent assay (IFA) and Western blot. Conclusion: These results can provide a potential value for structural and functional studies of EMCV-3AB and early diagnosis of Encephalomyocarditis virus infection.

Animals ; Hepatocyte Growth Factor ; metabolism ; Humans ; Proto-Oncogene Proteins c-met ; Serine Endopeptidases

OBJECTIVETo observe the efficacy and safety of modified Shengma Biejia Decoction (MSBD) combined with CAG program in treating elderly acute myeloid leukemia (AML) patients with yin deficiency toxin stasis syndrome (YDTSS).

METHODSTotally 46 elderly AML patients were assigned to the treatment group (24 cases; treated with MSBD + CAG) and the control group (22 cases; treated with CAG + placebos of Chinese medicine) according to random digit table. The therapeutic course of CM placebo or MSBD was 21 days. The clinical efficacy and adverse reactions were observed. Meanwhile, physical state (ECOG Score), transfusion dependency, and TCM syndrome score were compared before and after treatment.

RESULTS(1) The complete remission rate was 54% (13/24) and the objective response rate (ORR) was 71% (17/24) in the treatment group, obviously higher than those of the control group [36% (8/22); 54% (13/24)], with statistical difference (P = 0.036, 0.042). When comparing the efficacy based on risk level, the moderate and poor ORR was 71% (10/14) and 67% (6/9) in the treatment group, and 57% (8/14) and 33% (2/6) in the control group, with statistical difference between the two groups (P = 0.048; P = 0.010). (2) Compared with before treatment in the same group, the ECOG score significantly decreased, the average infusion time of red cells and platelets were markedly prolonged in the treatment group after treatment (P < 0.05). ECOG score, the average infusion time of red cells and platelets were significantly better in the treatment group than in the control group after treatment (P < 0.05). (3) Compared with before treatment in the same group, scores of fever, hemorrhage, and bone pain were markedly reduced in the control group (P < 0.05); scores of fever, fatigue, hemorrhage, dry mouth, and bone pain were markedly reduced in the treatment group (P < 0.05). Better effect in relief of fever, fatigue, hemorrhage, dry mouth, and so on was obtained in the treatment group than in the control group (P < 0.05). (4) In aspect of hematotoxicity, the incidence of neutropenia, anemia, thrombocytopenia was obviously lower in the treatment group than in the control group [29.2% (7/24) vs 54.5% (12/22); 16.7% (4/ 24) vs 45.5% (10/22); 33.3% (8/24) vs 63.6% (14/22); P < 0.05]. The incidence of fatigue and anorexia was obviously lower in the treatment group than in the control group [37.5% (9/24) vs 63.6% (14/22), 37.5% (9/24) vs 81.8% (18/22); P < 0.05].

CONCLUSIONMSBD combined with CAG program in treating elderly AML patients with YDTSS, with efficacy enhancing toxicity reducing effect, had distinct advantages in improving physical condition and clinical symptoms, and reducing transfusion dependency.

Aclarubicin ; therapeutic use ; Aged ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Cytarabine ; therapeutic use ; Drugs, Chinese Herbal ; therapeutic use ; Granulocyte Colony-Stimulating Factor ; therapeutic use ; Humans ; Leukemia, Myeloid, Acute ; drug therapy ; Medicine, Chinese Traditional ; Phytotherapy ; Yin Deficiency ; drug therapy

OBJECTIVETo explore the dose-effect relationship between premature chromosome condensation induced by Calyculin A and irradiation dose.

METHODSThe human peripheral blood was irradiated by (137)Cs gamma radial. The irradiation dose included 0, 0.5, 1.0, 2.0, 3.0, 4.0, 5.0 Gy. The premature chromosome condensation induced by Calyculin A was observed, and dyed by centromeric banding.

RESULTSThere was the quadratic relation between the total aberration, fragment, dicentric+centric ring (dic+r) ration and irradiation dose.

CONCLUSIONPremature chromosome condensation induced by Calyculin A can be used as a biodosimetry.

Cell Line ; Chromosome Aberrations ; radiation effects ; Chromosome Banding ; Chromosomes ; radiation effects ; Dose-Response Relationship, Radiation ; Female ; Gamma Rays ; adverse effects ; Humans ; Lymphocytes ; radiation effects ; Male ; Oxazoles ; pharmacology ; Young Adult

OBJECTIVETo analyze the sectional anatomical features of auricular and middle ear malformation in patients with microtia so as to improve the clinical classification and the instruction of surgery.

METHODSFrom Jun. to Dec. 2009, 36 cases with microtia were selected in the center of auricular reconstruction in Plastic Surgery Hospital, including 22 cases of unilateral microtia and 14 cases of bilateral microtia. 22 patients with unilateral microtia were studied with the contralateral healthy ears as controls. Spiral CT was performed for high-resolution scan of the temporal bone. The coronal, sagittal and 3D reconstruction images were created with Mimic software. Several distances and degrees were measured.

RESULTSThe patients were classified by Max classification. The anteroposterior diameter and the vertical diameter of tympanic cavity were (7.75 +/- 1.92) mm and (14.66 +/- 4.75) mm for type I; (6.17 +/- 2.56) mm and(14.35 +/- 5.12) mm for type II; (6.31 +/- 3.40) mm and (9.97 +/- 4.36) mm for type III (P = 0.001). The mastoid pneumatization degree for type I, II, III were 13.33%, 13.64%, 30.77% in sclerotic type, 13.33%, 18.18%, 7.69% in diploe type, 0, 9.09%, 38.46% in composite type, 73.33%, 59.09%, 23.08% in pneumatic type (chi2 = 24.11, P = 0.002). The cover of fenestra vestibuli by facial nerve was 21.43%, 47.62%, 54.55% (chi2 = 23.44, P = 0.002) for type I, II, III. There was a statistical difference between the microtia group and the control group.

CONCLUSIONSAccording to the Max classification, the middle ear malformation changed along the auricular malformation. The anatomical variations was complicated in type II microtia, which should be sub-classified.

Adolescent ; Adult ; Anatomy, Cross-Sectional ; Child ; Child, Preschool ; Ear ; abnormalities ; diagnostic imaging ; Ear, External ; abnormalities ; diagnostic imaging ; Ear, Middle ; abnormalities ; diagnostic imaging ; Female ; Humans ; Male ; Tomography, Spiral Computed ; Young Adult

OBJECTIVETo investigate the role of activated hepatocyte growth factor (HGF) in apoptosis of hepatic stellate cells (HSCs) and in modulating the Rho signaling pathway.

METHODSHSCs were divided into the following groups: blank control, consisting of HSCs without treatment; two treatment controls, consisting of HSCs exposed to exogenous HGF at 50 ng/ml and HSCs exposed to exogenous HGF activator (HGFA) at 70 ng/ml; three experimental groups, consisting of HSCs exposed to both exogenous HGF and HGFA, HSCs pre-incubated with the HGF inhibitor c-met at 500 ng/ml for 6 hours and then exposed to exogenous HGF and HGFA, and HSCs pre-incubated with the Rho pathway inhibitor Y-27632 at 10 ng/ml and then exposed to exogenous HGF and HGFA. Activation status of the cultured HSCs was determined by change in expression of alpha-smooth muscle actin (SMA). The optimal intervention concentration of Y-27632 was determined by MTT assay. The apoptotic status of HSCs was determined by flow cytometry. Expression of the HGF-alpha chain was detected by immunofluorescence. The expression of RhoA was evaluated by PCR (for mRNA) and by immunohistochemical staining and Western blot analysis (for protein).

RESULTSExposure to 10 mumol/L Y-27632 led to obvious growth inhibition of HGF + HGFA-induced HSCs, compared with the other concentrations tested (P less than 0.05). HGF + HGFA induced the expression of the HGF-alpha chain in a time-dependent manner (P less than 0.01); however, the increases in expression of HGF-alpha chain induced by HGF alone and HGFA alone were not significantly different from the level in the blank controls (P more than 0.05). Exposure to HGF alone and HGFA alone led to a time-dependent increase in apoptosis (24 h, 48 h, 72 h) but exposure to HGF + HGFA led to the highest levels of apoptosis (P less than 0.05). Exposure to HGF + HGFA led to a time-dependent decrease in RhoA mRNA and protein expression (P less than 0.01).

CONCLUSIONActivation of hepatocyte growth factor promotes apoptosis of hepatic stellate cells by suppressing RhoA expression and down-regulating the Rho signaling pathway.

OBJECTIVETo classify the blood vessel distribution of the expanded skin in the mastoid region and its relevance to reasonable and reliable design of the expanded flap for auricular reconstruction in congenital microtia.

METHODSThe blood vessel distribution of the expanded skin in the mastoid region was observed by light permeation test. The expanded flaps with different ratio of length to width were designed according to their blood vessel distribution types.

RESULTSThe vascular distribution of the expanded skin in 403 cases was divided into five types. All the flaps survived completely.

CONCLUSIONSThe blood vessel distribution type of the expanded skin in mastoid region has great significance for the design of post-auricular expanded flap in auricular reconstruction.

Congenital Abnormalities ; surgery ; Dermatologic Surgical Procedures ; Ear ; abnormalities ; Female ; Humans ; Male ; Mastoid ; surgery ; Skin ; blood supply ; Skin Transplantation ; Surgical Flaps ; blood supply ; Tissue Expansion Devices

OBJECTIVETo evaluate the impacts of Helicobacter pylori (H. pylori) infection on atherosclerosis and plasma lipid levels in high-cholesterol diet fed C57BL/6 mice.

METHODFemale C57BL/6 mice were randomly divided into 4 groups (n = 12 each): fed with normal chow diet (A), infected with H. pylori (B), fed with high-cholesterol diet (C) and infected with H. pylori and fed with high-cholesterol diet (D). After 52 weeks, plasma levels of lipids were measured and aortic atherosclerosis was observed. The ureA, ureC, cagA and vacA DNA were also detected by PCR in the aortic arteries.

RESULT(1) Prevalence of atherosclerosis was similar between group C and D (91.6% vs. 100%, P > 0.05) while there was no atherosclerosis in group A and B. H. pylori infected mice showed more obvious inflammation in gastric mucosa than mice without H. pylori infection. (2) The plasma levels of triglyceride, total cholesterol and LDL were higher and HDL was lower in group B, C and D than those in group A and in group D than in group C (all P < 0.05). (3) Roberts & Thompson scores and number of foam cells in plaques were significantly higher in group D compared with those in group C (all P < 0.05). (4) ureC DNA was detected in 5 out of 12 aortic arteries of mice in group D but not in group A, B and C.

CONCLUSIONOur results suggested that H. pylori infection might enhance the atherosclerotic lesion formation in this mouse model.

Animals ; Atherosclerosis ; microbiology ; pathology ; Bacterial Proteins ; genetics ; Bacterial Toxins ; genetics ; Cholesterol ; blood ; Cholesterol, Dietary ; adverse effects ; DNA, Bacterial ; analysis ; Female ; Helicobacter Infections ; pathology ; Helicobacter pylori ; genetics ; Mice ; Mice, Inbred C57BL ; Triglycerides ; blood ; Urease ; genetics

OBJECTIVETo explore the application of the remnant in repairing the reconstructed ear in microtia.

METHODSFrom October 2000 to July 2006, 1,823 patients with reconstructed ear underwent the second operation for repairing. The remnants were used to reconstruct tragus, antitragus, crus of helix, concha cavum, or relax the adhesion between the reconstructed auricle and cranio wall, or insert into the reconstructed lobule or under the basement of the costal cartilage frame to restore the cranio-auricular angle or to the preauricular subcutaneous to correct the facial local depression.

RESULTSIn 1,823 cases, the tragus, antitragus and concha cavum was reconstructeded by remnant auricle in 1,180 cases; helix crus was repaired by remnant auricle in 743 cases; the wound was repaired by remnant auricle flap resulted from relaxing the adhesion between the reconstructed auricle and cranial wall in 68 cases; Subcutaneous and cartilage flap was formed and transferred in 95 cases, and among them the flap was transferred under the basement of the frame to restore the cranio-auricular angle in 77 cases and was transferred to the preauricular subcutaneous to correct the facial local depression in 18 cases; the skin flap was formed by remnant auricle to repair the lobule in 37 cases. All flaps survived completely. The appearance of reconstructed ear was satisfied.

CONCLUSIONThe remnant is very useful in repairing the reconstructed ear in microtia. It should be preserved propriately in auricular reconstruction.

Adolescent ; Adult ; Cartilage ; transplantation ; Child ; Child, Preschool ; Ear ; abnormalities ; surgery ; Ear, External ; abnormalities ; surgery ; Female ; Humans ; Male ; Reconstructive Surgical Procedures ; methods ; Ribs ; transplantation ; Surgical Flaps ; Suture Techniques ; Young Adult