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.

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

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

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 chest contour deformities after harvesting of costal cartilage for total ear reconstruction, so as to prevent donor site morbidity.

METHODS107 cases of microtia who underwent ear reconstruction with costal cartilage during the period from 1994 to 2003, were retrospectively studied. Both the chest depression deformity and the costal arch deformity were evaluated. The possible related factors to the chest contour deformities were analyzed statistically with Stata software.

RESULTSThe incidence of chest depression was 54.55% (48/88), including 35 cases of mild depression (39.77%) and 13 cases of severe depression (14.77% ). The incidence of costal arch deformity was 52.77% (46/88), including 33 cases of mild deformity (37.50%) and 13 cases of severe deformity (14.77%). The incidence of both deformities was significantly different at different age (P < 0.01), but not at different follow-up period (P = 0.294 and P = 0.225, respectively). Gender was related to the incidence of chest depression (P < 0.01), but not to the costal arch contour deformity (P = 0.619). Comparing resection of the 6th and 7th rib cartilage with resection of 7th and 8th rib cartilage, the incidence of costal arch deformity was significantly different (P < 0.01), but the incidence of depression deformity was not (P = 0.657).

CONCLUSIONSThe incidence of chest deformities increases with early operation time (< 13 years old). The male patients have higher incidence of depression deformity than female. The arch deformity is higher when the 6th and 7th rib cartilage were resected. The follow-up period is not related to the chest deformities.

Adolescent ; Adult ; Child ; Child, Preschool ; Ear, External ; surgery ; Female ; Follow-Up Studies ; Humans ; Male ; Postoperative Complications ; Reconstructive Surgical Procedures ; adverse effects ; Retrospective Studies ; Ribs ; abnormalities ; transplantation ; Sex Factors ; Thorax ; abnormalities ; Young Adult

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

OBJECTIVETo classify and repair "Butterfly Ear" deformity which presents characters of dysplasia of inferior auricle of ear and congenital bat ear.

METHODThe repairment procedures include: type I: auricular cartilage flap inversion folding technique. type II: local ear skin flap. type III: soft tissue expander autogenous, rib cartilage framework.

RESULTSThe method was used in 19 cases from October 2001 to March 2005. Postoperative follow-up showed satisfactory results in all cases.

CONCLUSIONAccording to "Butterfly Ear" deformity classification, different technique could be applied.

Adolescent ; Adult ; Child ; Congenital Abnormalities ; classification ; surgery ; Ear, External ; abnormalities ; surgery ; Female ; Humans ; Male ; Reconstructive Surgical Procedures ; methods ; Surgical Flaps ; Young Adult

OBJECTIVETo investigate the effect of sphingosine kinase 1 (SphK1) on the proliferation, apoptosis, migration and invasion of colon cancer TH-29 cells and to explore its molecular mechanisms.

METHODSPhorbol 12-myristate 13-acetate (PMA) was used to induce the activity of SphK1 and N, N-dimethylsphingosine (DMS) was used to suppress the activity of SphK1. Cell prolieration and apoptosis were detected by MTT assay and flow cytometry, respectively. The migration and invasion capabilities of the cells were assessed in Transwell chambers. The activity of SphK1 was assayed by autoradiography. Western blot was used to evaluate the protein expression of SphK1, p38, phosphorylated p38 (p-p38) and SAPK/JNK.

RESULTSPMA and DMS were able to induce and suppress the activity and protein expression of SphK1 in a time-dependent manner, respectively. PMA enhanced and DMS suppressed the cell viability in a time- and dose-dependent manner. Being treated with 100 nmol/L PMA or 50 µmol/L DMS for 0, 6, 12, 24 h, the cell apoptosis rates of PMA group were (9.35 ± 0.84)%, (7.61 ± 0.48)%, (5.53 ± 0.76)% and (0.56 ± 0.33)%, contrastly, that of DMS group were (9.18 ± 0.94)%, (12.06 ± 1.41)%, (19.80 ± 2.36)% and (31.85 ± 3.60)%, respectively. Compared with the control group, the cell migration and invasion capabilities of the PMA group were significantly enhanced, and that of the DMS group were significantly suppressed. The migration cell number of control, PMA and DMS groups were 68.75 ± 6.15, 109.33 ± 11.63 and 10.83 ± 2.48, the invasion cell number of control, PMA and DMS groups were 55.42 ± 4.50, 90.58 ± 7.06 and 9.58 ± 2.39, respectively. With the elevating activity and expression of SphK1, the protein expressions of p38, p-p38 and SAPK/JNK were strikingly suppressed. On the contrary, after treating with DMS the protein expressions of p38, p-p38 and SAPK/JNK were enhanced.

CONCLUSIONSSphK1 potently enhances the prolieration, migration and invasion of colon cancer HT-29 cells, meanwhile suppresses the cell apoptosis. The suppressing of the p38 and SAPK/JNK signalling pathways may be one of its molecular mechanisms.

Apoptosis ; drug effects ; Carcinogens ; administration & dosage ; pharmacology ; Cell Movement ; drug effects ; Cell Proliferation ; drug effects ; Dose-Response Relationship, Drug ; Enzyme Inhibitors ; administration & dosage ; pharmacology ; HT29 Cells ; Humans ; MAP Kinase Kinase 4 ; metabolism ; Neoplasm Invasiveness ; Phosphorylation ; Phosphotransferases (Alcohol Group Acceptor) ; metabolism ; physiology ; Sphingosine ; administration & dosage ; analogs & derivatives ; pharmacology ; Tetradecanoylphorbol Acetate ; administration & dosage ; analogs & derivatives ; pharmacology ; Time Factors ; p38 Mitogen-Activated Protein Kinases ; metabolism