A rapid detection of foot-and-mouth disease virus (FMDV) was established by using reverse transcription loop-mediated isothermal amplification (RT-LAMP) method, meanwhile its specificity and sensitivity were assessed. The results showed that the FMDV RNA could be amplified by incubation at 65degrees C for only 1h using six primers designed based on FMDV polyprotein gene and the amplification products could be detected easily by naked-eye. There is no cross reaction with other virus such as SVDV, SFV and PPV by detecting their RNA samples. The detection limit of this method was found to be 10(-5) dilution of RNA sample which was 100-fold higher than that of PCR and 10-fold higher than real-time PCR.
Animals ; DNA Primers ; Foot-and-Mouth Disease ; prevention & control ; virology ; Foot-and-Mouth Disease Virus ; genetics ; isolation & purification ; Genome, Viral ; genetics ; Nucleic Acid Amplification Techniques ; RNA, Viral ; Reverse Transcriptase Polymerase Chain Reaction ; methods ; Sensitivity and Specificity ; Swine Vesicular Disease ; virology

OBJECTIVETo explore the feasibility of evaluating complete ischemia-reperfusion injury (IRI) of the testis by contrast-enhanced ultrasonography with microbubbles (MB) targeted to P-selectin (MBp) in rabbits.

METHODSWe randomly divided 30 healthy adult rabbits into five groups of equal number (control, 0.5 h IRI, 1 h IRI, 2 h IRI, and 4 h IRI), prepared phospholipid MB and MBp, and performed contrast-enhanced ultrasonography of the bilateral testes with MB or MBp at an interval of 20 min at different times after IRI. When MB or MBp disappeared completely in the healthy testis at 4 to 5 min after intravenous injection, we recorded the power of the first frame (F-P) in the IRI testes followed by immunohistochemical staining of the testis tissue.

RESULTSCEU with MBp achieved a significantly higher F-P than that with MB in all the IRI groups (P < 0.05), which was (8.34 +/- 1.20) versus (1.87 +/- 0.25) 10(-5) AU at 2 hours, but there was no significant difference between MB and MBp in the control rabbits (0 AU, P > 0.05). Immunohistochemistry showed a significantly time-dependent increase in the expression of P-selectin in the vascular endothelial cells of the IRI testes, but not in those of the control.

CONCLUSIONContrast-enhanced ultrasonography with MBp can be used to evaluate the inflammatory reaction of testicular ischemia-reperfusion injury.

Animals ; Antibodies ; Disease Models, Animal ; Male ; Microbubbles ; P-Selectin ; immunology ; Rabbits ; Reperfusion Injury ; diagnostic imaging ; Testis ; blood supply ; Ultrasonography

This study was aimed to investigate the expressions of tumor necrosis factor-alpha (TNF-alpha), Interleukin-6 (IL-6) in serum and the incidence of multiple organ dysfunction syndrome (MODS) in pigs with hemorrhagic shock after the blood transfusion simultaneously combined with different doses of free hemoglobin (FHb) so as to provide guidance of banked blood with high concentration of FHb during war injury through understanding effect of FHb on the animals. The different doses of FHb were given intravenously during the recovery of pig from shock, the vital signs and functional changes of vital organs were monitored and the incidence of MODS was determined, as well as the serum specimens were collected and the TNF-alpha, IL-6 levels in serum were detected by ELISA. The results showed that there were statistical differences of serum levels of TNF-alpha and IL-6 in pigs after FHb 10 mg/kg infusion, as compared to shock control group. There was significantly difference of the serum levels of TNF-alpha, IL-6 after FHb 15 mg/kg infusion, compared to the control group. The incidence of MODS increased significantly. It is concluded that the blood infusion containing high dose (more than 10 mg/kg) of FHb influences significantly on the cytokines in pigs with hemorrhagic shock, and increases damage of cytokines to vital organs and the incidence of MODS. The tolerance dose of the pigs to free hemoglobin is about 10 mg/kg or so. The infusion of blood with less than 10 mg/kg is relatively safe for pig in hemorrhagic shock.
Animals ; Disease Models, Animal ; Hemoglobins ; analysis ; Interleukin-6 ; blood ; Multiple Organ Failure ; etiology ; Serum ; metabolism ; Shock, Hemorrhagic ; blood ; Swine ; Tumor Necrosis Factor-alpha ; blood

OBJECTIVETo construct an inducible eukaryotic vector carrying red fluorescent protein (DsRed) and evaluate the regulation of DsRed gene expression in vitro.

METHODSThe vector pRS17-RUDsRed containing DsRed gene, promoter and RU486-inducible system was constructed using molecular biological methods. To minimize potential interference, the two transcriptional elements were spaced with a 1.6 kb insulator. Fluorescence microscopy and flow cytometry were used to observe the activation of this regulatable vector after transfection in MFC cells.

RESULTSThe vector was identified by digestion with different restriction enzymes, sequencing and PCR. In the absence of RU486, the cells transfected with the vector exhibited very low DsRed protein expression, and the addition of RU486 induced efficient DsRed expression in the cells.

CONCLUSIONThe RU486-inducible eukaryotic vector carrying DsRed protein allows effective regulation of the target gene expression in vitro, which provides a useful tool for gene regulation and gene therapy studies.

Gene Expression Regulation ; Genetic Therapy ; methods ; Genetic Vectors ; genetics ; Humans ; Luminescent Proteins ; genetics ; metabolism ; Mifepristone ; pharmacology ; Promoter Regions, Genetic ; genetics ; Stomach Neoplasms ; genetics ; pathology ; Tumor Cells, Cultured

OBJECTIVETo study the expression and localization of co-stimulators in the mucosa of patients with ulcerative colitis (UC), and to explore its role in the pathogenesis of UC.

METHODSExpression of co-stimulators CD86 and inducible co-stimulator (ICOS) was studied by immunohistochemistry on paraffin-embedded mucosal tissue from patients with active UC (64 cases), inactive UC (51 cases) and normal controls (20 cases). Immunostaining for CD28 was also carried out on frozen fresh mucosal tissue sampled from patients with active UC (7 cases), inactive UC (2 cases) and normal controls (5 cases). In addition, expression of CD4, CD8 and CD20 were also examined.

RESULTSIn active UC, increased expression of CD86 was not only observed in lamina propria mononuclear cells but also in the intestinal epithelial cells, as compared with inactive UC and the normal controls (P < 0.01). Increased ICOS expression in lamina propria mononuclear cells was detected in active UC, as compared with inactive UC and the normal controls (P < 0.01). Increased ICOS expression in intestinal epithelial cells was also seen in active UC, as compared with that of inactive UC (P < 0.01). The expression of CD86 was higher in inactive UC than in the normal controls (P < 0.05 or P < 0.01). However, the expression of ICOS showed no statistically significant difference between inactive UC and normal controls. Increased expression of CD28 in active UC, compared with that in inactive UC and normal controls, was also noticed (P < 0.05 or P < 0.01). The number of CD4 or CD8-positive intraepithelial lymphocytes and lymphocytes infiltrating in the lamina propria and small vessel walls was much higher in active UC than in inactive UC and normal controls (P < 0.01). Moreover, the ratio of CD4/CD8 was highest in active UC (P < 0.01). The number of CD20-positive B lymphocytes in lamina propria was also higher in active UC than in inactive UC and normal controls (P < 0.01).

CONCLUSIONSIn active UC, CD86 and ICOS were over-expressed in the intestinal epithelial cells and lamina propria mononuclear cells. The phenomenon suggests that abnormal expression of co-stimulators may contribute to the deregulation of acquired immune responses in UC.

Adult ; Aged ; Antigens, Differentiation, T-Lymphocyte ; metabolism ; B7-2 Antigen ; metabolism ; CD28 Antigens ; metabolism ; CD4-CD8 Ratio ; Case-Control Studies ; Colitis, Ulcerative ; metabolism ; pathology ; Epithelial Cells ; metabolism ; pathology ; Female ; Humans ; Immunohistochemistry ; Inducible T-Cell Co-Stimulator Protein ; Intestinal Mucosa ; metabolism ; pathology ; Leukocytes, Mononuclear ; metabolism ; pathology ; Male ; Middle Aged ; Mucous Membrane ; metabolism ; pathology ; Young Adult

Objective To analyze the therapeutic effect and prognosis according to a new CT classification of traumatic posterior occipital epidural hematomas (POEH). Methods We classified the CT presentation of 104 patients with POEH by sinus transverses: type Ⅰ hematomas were defined as up-sinus transverses; type Ⅱ hematomas were defined as sub-sinus transverses and type Ⅲ hematomas were defined as straddling sinus transverses. The above types were divided into unilateral or bilateral subtypes. Bone flap craniotomy through a median posterior approach was performed in patients with unilateral hematoma adjacent to the midline. Bone flap craniotomy through a median suboccipital approach was adopted for patients with bilateral middle occipital hematoma. Results Type Ⅰ enjoyed mild symptoms, good curative effect and low mortality rate (7%). Type Ⅱ and Ⅲ had serious symptoms and high mortality rate (13.3%, 16.6%, respectively). Conclusion The classification of the CT presentation of POEH is helpful in confirming the diagnosis, drawing up the scheme of treatment and judging the prognosis.

Carcinoma, Hepatocellular ; blood ; complications ; virology ; Glycomics ; Hepatitis B virus ; Hepatitis B, Chronic ; blood ; Humans ; Liver Cirrhosis ; blood ; complications ; virology ; Liver Neoplasms ; blood ; complications ; virology ; Neoplasm Staging

OBJECTIVETo investigate the effect of replication-competent adenovirus-mediated interleukin-12 gene to chemotherapeutic sensitivity on gastric cancer cell.

METHODSReplication-competent adenovirus and replication-competent adenovirus- mediated interleukin- 12 gene was constructed and expanded separately. The mortality of gastric cancer cell caused by the CNHK200- mIL- 12, Onyx- 015 in combination with different dosages of chemotherapeutic agents were evaluated by MTT assay at the same viral titer with a series of different dosages of chemotherapeutic agent,or at a series of different viral titers with the same dosage of chemotherapeutic agent. The curative effect to the xenografts gastric tumor in nude mouse was also observed by two viruses solely or together with 5-Fu.

RESULTSThe lytic activity of replication-competent adenovirus to gastric cancer cell line SGC-7901 was relatively poor at MOI value of 0.5, but it could be improved significantly when combined with chemotherapeutic agents of ADM, 5-Fu or CAP compared to the simple chemical therapy (P< 0.05). Chemotherapeutic agent 5- Fu could not effectively kill SGC-7901 when used at a relatively low dosage of 10microg/ml,whereas its activity could be improved when combined with a replication-competent adenovirus,and the killing rate was much higher than that with replication-competent adenovirus solely (P< 0.05). The gastric tumor xenografts was prevented and killed by replication adenovirus solely or combined with 5-Fu.

CONCLUSIONThe replication- competent adenovirus- mediated interleukin- 12 gene can increase the chemotherapeutic sensitivity on gastric cancer cell. There is synergetic effect between the replication adenovirus and the chemotherapeutic agents in killing gastric cancer cell.

Adenoviridae ; genetics ; Animals ; Cell Line, Tumor ; Drug Screening Assays, Antitumor ; Genetic Therapy ; Humans ; Interleukin-12 ; genetics ; Male ; Mice ; Mice, Inbred BALB C ; Stomach Neoplasms ; drug therapy ; Viral Vaccines ; Virus Replication

OBJECTIVETo investigate the efficiency of damage control surgery (DCS) and predictors of mortality in critically multiple trauma patients.

METHODSFrom May 1998 to February 2007, DCS were carried out in 27 patients with critically multiple trauma. Of the patients 15 cases survived (survival group) and 12 cases died (dead group). The surgical complications, causes of death, demographic, physiologic and medical parameters were collected and compared between the two groups. Multiple logistic regression analysis were performed to identify possible predictors of mortality.

RESULTSThe incidence of surgical complications was 37.0 percent, and the intra-abdominal infections was the most frequent (18.5%). The overall mortality rate was 44.4 percent. The most common causes of death was multiple organ dysfunction syndrome (50.0%). With respect to predicting mortality, statistically significant differences was found in parameters as age, injury severity score (ISS), initial temperature and base excess (BE), estimated blood loss, initial ICU temperature and length of hospital stay. Older age, increased absolute value of initial BE and lower initial ICU temperature were determined as independent predictors of mortality on multiple logistic regression analysis.

CONCLUSIONSThere is a comparable high morbidity and mortality rate in severely injured patients managed with DCS. Increased age, a larger absolute value of initial BE and lower initial ICU temperature could independently predict death of the patients.

Adolescent ; Adult ; Age Factors ; Aged ; Female ; Humans ; Injury Severity Score ; Logistic Models ; Male ; Middle Aged ; Multiple Trauma ; mortality ; surgery ; Multivariate Analysis ; Postoperative Complications ; Prognosis ; Temperature ; Young Adult

OBJECTIVETo investigate the seroprevalence of anti-HAV IgG in adults of 4 cities in China.

METHODSSerum samples were collected from 2390 local residents aged between 20 to 88 years from Beijing, Shanghai, Wuhan and Guangzhou. The anti-HAV IgG in sera was detected with a microparticle enzyme immunoassay (MEIA).

RESULTSThe anti-HAV IgG seroprevalence in female of 30 to 39 years in Beijing (64.58%, 62/96) was higher than that in male (45.57% 36/79)) (x(2) = 6.358, P = 0.012). It increased with age in adults of Beijing and Guangzhou. The rates were 54.22 % (90/166), 56.00% (98/175) and 67.18% (88/131) for the 20-, 30- and 40-49 age groups in Beijing (x(2) = 4.76, P = 0.03); and 52.83% (56/106), 52.50% (63/120), 82.46% (94/114), 89.80% (88/98) and 96.77% (60/62) for the 20-, 30-, 40-, 50- and 60-88 age groups in Guangzhou, respectively (x(2) = 72.58, P less than 0.01). This trend was not found in Shanghai and Wuhan (x2 = 0.96, 2.99; P = 0.33, 0.08 respectively). The seroprevalence rates of anti-HAV IgG in the 20 to 39 age group of Beijing, Shanghai, Guangzhou and Wuhan were 55.13% (188/341), 63.93% (429/671), 52.65% (119/226) and 78.37% (308/393), respectively.

CONCLUSIONThe seroprevalence rates of anti-HAV IgG in young adults aged 20 to 39 years of the four cities are relatively low, and HAV vaccination should be suggested for the susceptible population of this age group in China.

Adult ; Age Distribution ; Aged ; Aged, 80 and over ; China ; epidemiology ; Female ; Hepatitis A ; epidemiology ; immunology ; prevention & control ; Hepatitis A Antibodies ; blood ; Hepatitis A Virus, Human ; immunology ; Humans ; Immunoglobulin G ; blood ; Male ; Middle Aged ; Seroepidemiologic Studies ; Sex Distribution ; Urban Population ; Young Adult