Objective　To construct the eukaryotic expression recombinant plasmid, pcIFN-γ, as a genetic adjuvant and observe the immune responses elicited by pcDNA3-rhoptry protein 1 （pc-ROP1） combined with pcIFN-γ against Toxoplasma gondii (T. gondii) infection in mice. Methods　A fragment of the IFN-γ gene was directly inserted into the pcDNA3 plasmid and identified by two restriction endonucleases digestion. pcIFN and pcROP1 DNA was injected into the left leg muscle of mice at a dosage of 100*!μg, and a booster vaccination was given at the same dosage after two weeks. Control groups were injected with pcDNA3 blank plasmid or normal saline. At 30, 50 and 70 days after booster injection, kinetic tests were carried out: MTT assay for the proliferation response of T lymphocyte cells and the activity of NK cells, sandwich ABC-ELISA for the determination of IFN-γ, IL-2 and IL-10; a serum enzymetic aassay for nitric oxide (NO) in sera and ELISA for the titer of IgG antibody in sera. Results　The recombinant plasmid, pcIFN-γ was constructed. The proliferation response of spleen T lymph cells, NK cell killing activity, and serum levels of IFN-γ, IL-2 and NO in mice injected with pcROP1 and pcIFN-γ were higher than in those injected with pcROP1 alone. There was no difference in IgG antibody levels between the two groups. Conclusion　The genetic adjuvant, pcIFN-γ, could enhance the cellular immune response induced by DNA vaccine of pcROP1 in mice against Toxoplasma gondii infection.

OBJECTIVETo characterize the immune response induced by SAG1 encoding plasmid combined with IL-2 gene adjuvant in mice and to assess the protective effect of this vaccination against toxoplasmosis.

METHODSMice were co-injected intramuscularly with plasmid encoding Toxoplasma gondii SAG1 plus murine IL-2 expression vector at a dose of 100 microg. Booster immunizations were employed 2 more times at 3-week interval. As controls, mice were inoculated with PBS or empty plasmid pcDNA3. Humoral and cellular responses were assayed using ELISA for the determination of Ab, Ab isotype and IFN-gamma, as well as IL-4. To detect the integration and dissemination of DNA in the injected mice, PCR and in situ hybridization were performed. All mice were then infected with highly virulent RH tachyzoites of Toxoplasma gondii intraperitoneally.

RESULTSSignificant increases in specific IgG levels were observed in mice after immunization three times with SAG1 expression plasmid. With respect to the IgG isotype, co-inoculation of IL-2 expression plasmid enhanced the level of IgG2a and the production of IFN-gamma. Challenging mice by vaccinating with combined plasmids with RH tachyzoites resulted in prolonged survival.

CONCLUSIONHumoral and cytokine responses elicited by SAG1 DNA immunization can be modulated by co-inoculation with IL-2 expression plasmid. The use of DNA vaccine in combination with an appropriate cytokine gene to prevent T. gondii infection warrants further investigation.

Animals ; Antibodies, Protozoan ; blood ; Antigens, Protozoan ; Cytokines ; biosynthesis ; Female ; Immunization ; Immunoglobulin G ; blood ; classification ; Interleukin-2 ; genetics ; Mice ; Protozoan Proteins ; genetics ; Protozoan Vaccines ; immunology ; Toxoplasma ; immunology ; Toxoplasmosis, Animal ; prevention & control ; Vaccines, DNA ; immunology

Objective To explore the expression and change of ski-interacting protein (SKIP) in rats after spinal cord injury. Methods A total of 60 adult female Sprague-Dawley rats were randomly divided into sham group (n=30) and spinal cord injury (SCI) group (n=30), each group was further divided into five time points including one day, three days, five days, seven days, and 14 days with six rats in each time points. The model was established at T10 with modified Allen's technique, and the sham group only bit the lamina of rats. The hindlimbs behavior was assessed with Basso-Beattie-Bresnahan (BBB) score at each time point. The pathological changes of spinal cord neurons were detected with Nissl staining. The expression of SKIP were observed with immunofluorescence staining. Results The BBB scores were signif-icantly lower in each time point in SCI group than in the sham group (t>48.267, P<0.001). Compared with the sham group, Nissl bodies in the cytoplasm of spinal cord neurons began to disintegrate, coalesce and irregularly distribute, the neurons began to degenerate and die on the fifth day, and the damage deteriorated on the 14th day. Immunofluorescence staining showed that SKIP expression was mainly expressed in the gray matter of the spinal cord and little expressed in the white matter. The expression of SKIP gradually increased after SCI, and reached a peak on the fifth day (t=-17.035, P<0.001) and decreased significantly on the 14th day (t=3.853, P<0.05). Conclusion SKIP may be a new signaling molecule, which play an important role in neuronal apoptosis after SCI.

OBJECTIVETo investigate the etiology and source of an infectious diarrhea outbreak and control the epidemic.

METHODSThrough the retrospective cohort study, we had surveyed all the residents who complained symptoms of diarrhea or vomiting since Nov. 20th,2007 from the five villages in the north of town Y, and collected hygiene information on the water supply system of the five villages, the environment information of three villages and hygiene information of some case-indexed families, and tested the etiological biomarker, including nucleoside acid of norovirus through Real-time PCR and nested PCR technologies.

RESULTSFrom Nov. 24th to Dec. 3th in 2007, 435 diarrhea or vomiting cases were found in the north of Y town, where tap water A was supplied for daily use. The attack rate was 12.93%. The diarrhea cases were distributed among all country groups who has used tap water A and the attack rate was ranged from 5.21% (20/384) to 21.23% (100/471). Drinking the tap water A was significantly associated with an increased risk of infection (RR = 9.246, 95% CI: 6.25 -13.68). About 85.9% (262/ 305) of the cases were from Nov. 25th to 27th. An investigation of a country of S2 group showed that the incidence of different age groups was distributed as the following: 0 - year-old 20.0% (3/15); 10 - year-old 17.3% (9/52); 20 - year-old 15.2% (16/105); older than 60 year-old 23.3% (7/30). No statistical significance was identified between age and infection(chi2 = 1.15, P >0.05). Most of the patients were not serious and well prognostic, and no hospitalized or dead cases were reported. On site investigation and daily water quality monitoring showed that disinfection procedures were not strictly followed. The monitoring data also indicated the bacteriology index of tap water A was disqualified. The test of Salmonella, Shigella and Staphylococcus aureus were negative in two vomit and one stool samples from patients. Three specimens by Real-time PCR, and six by nested PCR were positive for norovirus among the three feces and three anal swabs samples. With the drinking water sterilization and health education, the epidemic had been controlled rapidly and effectively.

CONCLUSIONThe epidemic was a diarrhea outbreak that might be caused by norovirus through drinking the contaminated tap water A.

Adolescent ; Adult ; Caliciviridae Infections ; epidemiology ; Child ; Child, Preschool ; Cohort Studies ; Diarrhea ; epidemiology ; virology ; Disease Outbreaks ; Humans ; Incidence ; Infant ; Infant, Newborn ; Middle Aged ; Retrospective Studies ; Water Pollution ; Water Supply ; Young Adult

OBJECTIVE: To explore the differences in Haversian system between human and animal bones through imaging analysis and morphology description. METHODS: Thirty-five slices grinding from human being as well as dog, pig, cow and sheep bones were observed to compare their structure, then were analysed with the researchful microscope. RESULTS: Plexiform bone or oeston band was not found in human bones; There were significant differences in the shape, size, location, density of Haversian system, between human and animal bones. The amount of Haversian lamella and diameter of central canal in human were the biggest; Significant differences in the central canal diameter and total area percentage between human and animal bones were shown by imaging analysis. CONCLUSION (1) Plexiform bone and osteon band could be the exclusive index in human bone; (2) There were significant differences in the structure of Haversian system between human and animal bones; (3) The percentage of central canals total area was valuable in species identification through imaging analysis.
Adult ; Animals ; Bone and Bones/ultrastructure* ; Cattle ; Dogs ; Haversian System/ultrastructure* ; Humans ; Image Processing, Computer-Assisted ; Microscopy, Electron ; Sheep ; Species Specificity ; Swine ; Tibia/ultrastructure*

OBJECTIVETo explore whether hepatitis C virus core protein (HCV C) regulates the expression of NFAT1 to participate in the progression and malignant biological behavior of intrahepatic cholangiocarcinoma cells.

METHODSThe recombinant plasmid pEGFP-N(3)-HCV C and the empty vector pEGFP-N(3) were cotransfected with enhanced green fluorescent protein (EGFP) into RBE cells using liposome. Real-time PCR and Western blotting were used to examine the expression of NFAT1 mRNA and protein in the transfected RBE cells. MTT assay was used to evaluate the changes in the cell proliferation, and the cell cycle changes were analyzed by flow cytometry.

RESULTSHCV C transfection significantly enhanced the expressions of NFAT1 mRNA and protein in RBE cells (P<0.05) and promoted the progression of cell cycle into G(2)/M phase to accelerate the cell proliferation.

CONCLUSIONTransfection with HCV C gene up-regulates NFAT1 expression and promotes the cell cycle progression and proliferation of intrahepatic cholangiocarcinoma cells, suggesting the involvement of HCV C in the progression of intrahepatic cholangiocarcinoma.

Bile Duct Neoplasms ; pathology ; Cell Cycle ; Cell Line, Tumor ; Cell Proliferation ; Cholangiocarcinoma ; pathology ; Gene Expression ; Humans ; NFATC Transcription Factors ; genetics ; Plasmids ; Transfection ; Viral Core Proteins ; genetics

OBJECTIVETo investigate the effects of shikonin on the proliferation, expression of CXCR4 and the migratory responses to CXCL12 in colorectal carcinoma cell line SW480.

METHODSThe proliferation of SW480 cells was assessed by MTT assay. Cell surface expression of CXCR4 was determined by flow cytometry. The migratory ability was determined by Transwell.

RESULTSShikonin inhibited the proliferation of SW480 cells in time- and concentration-dependent manner. The expression rate of CXCR4 in SW480 cells was 99.1%. After application of shikonin 0.01 micromol/L, 0.1 micromol/L and 1.0 micromol/L for 24 h, the expression rate of CXCR4 decreased to 76.0%, 59.1% and 35.5% respectively (F=1098.041, P <0.001), and the CXCL12-induced SW480 cell migratory inhibition rate was 25.2%, 38.5% and 55.7% respectively (F=48.970, P <0.001).

CONCLUSIONBesides having inhibiting tumor cell proliferation effect, Shikonin may also play a role in anti-metastasis via down-regulating the expression of CXCR4 and reducing the CXCL12-induced migratory response in colorectal carcinoma cell.

Cell Line, Tumor ; Cell Proliferation ; drug effects ; Chemokine CXCL12 ; metabolism ; Down-Regulation ; Humans ; Naphthoquinones ; pharmacology ; Receptors, CXCR4 ; metabolism

Alkaloids ; pharmacology ; Cisplatin ; pharmacology ; Fluorouracil ; pharmacology ; Hep G2 Cells ; Humans ; Quinolizines ; pharmacology ; Tumor Necrosis Factor Ligand Superfamily Member 13 ; metabolism

BACKGROUNDThe peripheral enhancement of small hepatocellular carcinoma (SHCC) is a rare appearance in dual phase images by helical computed tomography (CT). This study discusses this phenomenon and its correlative histopathology.

METHODSThe helical CT dual phase appearance of peripheral enhancement in SHCC was analyzed in 21 cases (22 lesions). All lesions were confirmed as SHCC by histopathological examination.

RESULTSIn these 22 lesions, enhanced peripheral ring in 20 lesions was incomplete, the thickness of enhanced peripheral ring varied and mural node could be found in hepatic arterial phase; only 2 lesions had complete peripheral ring enhancement and ring of uniform thickness in hepatic arterial phase. The enhancement of some peripheral rings and mural nodes dropped to very low density in portal venous phase. The tumour cells were grade I in 3 lesions, II in 16, III in 2 and IV in 1. The vascular supply was more abundant at the border than in the centre of 15 lesions and the vascular supply was deficient in both centre and border of the remaining 7 lesions. In 3 lesions, the pseudocapsule showed in the border of the lesion. In 12 lesions, flecks of necrosis were found in the border and/or centre of the lesion.

CONCLUSIONSThe characteristic peripheral enhancement in helical CT dual phase images of small hepatocellular carcinoma correlates with different vascular supplies, fibrous capsule and necrosis of the lesion.

Adolescent ; Adult ; Aged ; Carcinoma, Hepatocellular ; diagnostic imaging ; pathology ; Humans ; Liver Neoplasms ; diagnostic imaging ; pathology ; Male ; Middle Aged ; Tomography, Spiral Computed

OBJECTIVETo evaluate the result of laparoscopic colorectomy in treatment of colorectal cancer.

METHODSLaparoscopic colorectal surgery was performed in 78 patients with colorectal cancer. Operative procedures, complications and postoperative recovery were studied.

RESULTNone of the 78 patients died of laparoscopic colorectal surgery or complications. Eleven patients died from tumor metastasis and 2 from other causes. Twenty-one, 17, 8 patients for 1, 3, 5 years survived respectively. In nine patients who had received operation less than 1 year, no tumor recurrence or metastasis was found except in 1 patient 11 months after operation.

CONCLUSIONLaparoscopic colorectal cancer resection is essential to colectomy for colon and rectum cancer when indicated.

Adult ; Aged ; Aged, 80 and over ; Colectomy ; methods ; Colorectal Neoplasms ; mortality ; pathology ; surgery ; Female ; Humans ; Laparoscopy ; Male ; Middle Aged ; Neoplasm Staging ; Rectum ; surgery