Objective To study the action and mechanism of staphylococcal exfoliative toxin A(E-TA)on pemphigus foliaceus antigen(PFA)and pemphigus vulgaris antigen(PVA)expressed on cultured human keratinocytes.Methods Stratified human keratinocytes were incubated with ETA and then stained with sera from patients with pemphigus foliaceus or pemphigus vulgaris as the first antibodies and FITC-la-beled sheep anti-human IgG as the second antibody.Total protein was harvested from the cells pretreated with ETA and run on SDS-PAGE for Western blot with the same antibodies.Simultaneously,supernatants of the keratinocytes before and after ETA treatment were collected for detection of the levels of IL-1?,IL-6with ELISA kits.The caseinolytic activities of the supernatants were tested by spectrometry in which casein was used as a non-specific substrate.Results Down-expression of PFA was shown after ETA treatment while no change of PVA expression was found.The high intensity and continuous linear appearance of fluo-rescent staining before ETA treatment became weak and discontinuous after ETA treatment,which were re-covered gradually in24hours.The degradation of proteins recognized by PF sera after ETV treatment was revealed by Western blot.The decreasing tendency of IL-1?concentration was found in the supernatants of cell culture after ETA treatment,but IL-6level was too low to be detected.Increased caseinolytic activities were found in the supernatants,and declined36hours after ETA treatment.Conclusions ETA acts on PFA expressed on keratinocytes in vitro,which is reversible along with withdrawal of ETA.The mechanism of E-TA act on PFA may be related to proteolytic action instead of promoting cytokine secretion.

Objective To investigate the distribution of TH17 cells in peripheral blood of patients with rheumatoid arthritis(RA). Methods Intracelluar flow cytometirc detection of TH17 cells in peripheral blood was establised using PHA or PMA + Ion as stimulators in vitro. Thirty-five active and 30 stable RA pa-tients and sex and age paired healthy controls were enrolled in this study. Results The stimulating effect of PMA + Ion was better than that of PHA alone. Under PMA + Ion stimulation, the percentage of TH17 cells in peripheral blood from RA patients and healthy controls was increased significantly(P<0.05). With or with-out PMA + Ion stimulation, such percentage in active group was higher than that of stable group and both of them were higher than that of heathy controls(P<0.05). The distribution profile of TH1 cells was similar to that of TH17 cells. Conclusion There is a distribution abnormality of TH17 cells in peripheral blood of RA patients, which could be used as a new marker for the assessment of immunological condition in RA.

Objective To optimize the concentration of polyethylene glycol (PEG) for the growth of Chlamydia trachomatis (Ct) reference strains D-UW-5/Cx and E-UW-5/Cx,and to evaluate the effects of PEG on the sensitivity of Ct to 4 common antibiotics.Methods After the inoculation of Ct standard strains (D-UW-5/Cx and E-UW-5/Cx) into McCoy cell monolayer,different concentrations of PEG were added into the culture medium followed by centrifugation.After 2 hours of incubation at 37 ℃,the inoculum was removed and a complete culture medium containing 1 μg/mL cycloheximide was added followed by another 48-hour culture.Subsequently,the culture was fixed and subjected to iodine staining for the calculation of Ct inclusions and optimization of PEG for the growth of Ct.Some Ct standard strains were used to infect McCoy cells,with PEG (0.7％,wt/vol) added to the culture medium after inoculation and before centrifugation process,and when the infection rate reached higher than 90％,a microdilution method was utilized to evaluate the minimal inhibitory concentration (MIC) of 4 antibiotics,including azithromycin,minocyline,moxifloxacin and doxycycline.Thirty-one clinical specimens,which had been confirmed to be positive for Ct serovar D or E strain,were inoculated into McCoy cell monolayer for the passage of Ct with or without the presence of 0.7％ PEG.Results The optimal concentration of PEG was 0.7％ for the growth of Ct,and this concentration of PEG could increase the number of inclusion bodies of Ct serovar E by 3.44 folds,and that of Ct serovar D by 3.56 folds.In vitro,the MICs of the 4 antibiotics were consistent between PEG-treated and untreated Ct reference strains.Moreover,PEG notably increased the quantity of inclusion bodies of Ct serovar E or D from clinical specimens after passages.Conclusions PEG (0.7％) can enhance the growth of Ct serovar D and E,but has no obvious influence on antimicrobial susceptibility of Ct.

ObjectiveTo detect the changes of heat shock protein 60 (HSP60) during the subcultures of standard and clinical strains of Chlamydia trachomatis(Ct),and to determine if the absence of chlamydial inclusions is associated with the persistent infection of Ct.MethodsA total of 40 Ct strains isolated by cell culture from patients were included in this study and classified into 2 groups according to whether inclusions appeared after initial culture.Reverse transcription PCR was conducted to quantify the levels of HSP60 in specimens containing Ct after 1-4 subcuhures.Chi-squared test was performed to analyze the relationship between the levels of HSP60 and treatment outcomes of patients.ResultsThe levels of HSP60 in clinical specimens containing Ct serovar E were significantly higher in subcultures prior to the appearance of inclusions than in subcultures with the appearance of inclusions(P ＜ 0.05).The ratio of HSP60:16S rRNA mRNA expression after the first,second,third,fourth passage was 0.38 ± 0.06,0.39 ± 0.03,0.38 ± 0.04 and 0.39 ±0.03 respectively in 18 specimens with inclusions appearing after the initial culture,1.18 ± 0.10,0.28 ± 0.06,0.30 ± 0.03 and 0.29 ± 0.05 respectively in 12 specimens with inclusions appearing after the second culture,1.20 ± 0.04,1.20 ± 0.04,0.28 ± 0.04 and 0.28 ± 0.05 in 10 specimens with inclusions appearing after the third culture.Whether inclusions appeared after the initial culture was associated with the treatment outcome of patients.Inclusions were undetected after the initial culture in 16 of 20 specimens from patients with poor response to treatment,but observed in 14 of 20 specimens from patients who tested negative for Ct after one course of treatment.Conclusions It is implicated that no inclusions form after the initial culture in 80％ of specimens from patients experiencing treatment failure.The Ct strains whose inclusions do not form after passages may be in a persistent state,and the expression of HSP60 is high in these strains.Specimens should be subjected to at least 3 blind passages to avoid missed diagnosis of Ct infection.

A new thermophilic fungus J18 isolated from the soil samples was identified as Paecilomyces thermophila. This strain produced effectively xylanase utilizing several lignocellulosic materials in the solid-state fermentation (SSF) , and wheat straw was the best carbon source. The results of single-factor-experiment showed that the wheat straw of particle size 0. 3 mm ~ 0.45 mm, initial moisture content of 83% , initial pH of 7. 0 and cultivation temperature of 50℃were the optimal conditions for xylanase production. Under the optimized conditions, it produced 18 580 U/g dry substrate after 8 days of cultivation. Therefore, xylanase production by Paecilomyces thermophila in SSF possess great potential for commercial applications.

OBJECTIVETo investigate the relation of the viral markers in serum and those expressed by hepatocytes to pathological lesions of hepatic tissue in patients with chronic hepatitis B.

METHODSThe relation of viral markers including HBsAg, HBsAb, HBeAg, HBeAb, HBcAb and HBV DNA in serum of 647 patients with chronic hepatitis B and HBsAg, HBcAg expressed by hepatocytes in 418 of these patients to pathological lesions of hepatic tissue was determined.

RESULTSViral markers in serum and those expressed by hepatocytes in patients with chronic hepatitis B were closely correlated with pathological lesions of hepatic tissue.

CONCLUSIONThe degree of inflammation and fibrosis in hepatic tissue is milder in serum HBsAg, HBeAb, HBcAb positive and HBV DNA negative patients but more serious in those with negative hepatocytic expression of HBsAg and HBcAg. HBV DNA is not significantly associated with pathological lesions of hepatic tissue.

Adolescent ; Adult ; Child ; Child, Preschool ; DNA, Viral ; blood ; genetics ; Female ; Hepatitis B Core Antigens ; blood ; Hepatitis B Surface Antigens ; blood ; Hepatitis B e Antigens ; blood ; Hepatitis B virus ; genetics ; immunology ; physiology ; Hepatitis B, Chronic ; blood ; pathology ; virology ; Host-Pathogen Interactions ; Humans ; Infant ; Infant, Newborn ; Liver ; pathology ; virology ; Male ; Middle Aged ; Young Adult

Objective To evaluate the clinical feasibility of localization CT enhanced image replacing plain CT scan image for target delineation and dose calculation.Methods Forty cases of NPC were collected and divided into two groups with different concentrations of contrast agents.The contours of planning target volume (PTV) and organs at risk (OARs) of each case were delineated in the plain scan image,and the contours of PTV and OARs were copied to the enhanced image.Two plans based on the plain scan image and the enhanced image were designed in the planning system of Eclipse.The dose distribution and OARs and MU were compared between the groups.Results No statistical differences were found in the dosimetry of PTV,OARs and MU (P＞0.05).Conclusion The image intensifier has little effect on the dose calculation of Eclipse for NPC.In the radiotherapy for NPC,the localization CT enhanced image can be used to replace the plain CT scan image for target delineation and dose calculation.

OBJECTIVETo identify novel human gastric cancer-associated susceptibility gene for early diagnosis and treatment of gastric cancer.

METHODSA primer was designed for 3'-rapid amplification of cDNA end(RACE) and amplified fragments were cloned, then they were analyzed by sequencing. Compared with ESTs in Genbank, the EST fragment represented a novel gene. Combination of Northern blot and virtual Northern and multiple tissues Northern blot, expression of the cDNA in multiple normal and carcinoma tissues were analyzed.

RESULTSOne of the important cDNA bands with poly(A) tail was cloned. This band was named W41. Sequence analysis showed that W41 consists of 533 bp. Basic local alignment search tool analysis revealed that W41 has low identity with any genes from GenBank. This sequence data was submitted to GenBank with accession No. AF 325202. Northern blot revealed that W41 presented higher expression in gastric cancer tissue than in normal tissue. Multiple tissue Northern blot revealed that W41 presented higher expression in multiple cancers than in normal tissues. Virtual Northern revealed that the cDNA presented higher expression in tumor series analysis of gene expression libraries than in normal.

CONCLUSIONA novel human gastric cancer-associated cDNA fragment was identified.

Blotting, Northern ; Cloning, Molecular ; DNA, Complementary ; chemistry ; genetics ; Female ; Gene Expression Regulation, Neoplastic ; Genetic Predisposition to Disease ; genetics ; HL-60 Cells ; HeLa Cells ; Humans ; K562 Cells ; Molecular Sequence Data ; Sequence Analysis, DNA ; Stomach Neoplasms ; genetics ; Tumor Cells, Cultured

Objective:To detect the proportion of lymphocyte subsets in peripheral blood of the ad-vanced breast cancer patients before and after chemotherapy with docetaxel and thiotepa,as well as the association between the proportion of peripheral blood lymphocyte subsets with the response rate and prog-nosis.Methods:The proportions of lymphocyte subsets (CD3 +T cell,CD3 +/CD4 +T cell,CD3 +/CD8 +T cell,CD3 -/CD16 +56 +NK cell,CD3 +/CD16 +56 +T cell,CD19 +B cell,CD4 +/CD25 +T cell,CD8 +/CD28 -T cell,CD8 +/CD28 +T cell)in the peripheral blood of 86 patients were analyzed with flowcytometry before and after chemotherapy.The result was analyzed in combination with clinico-pathological data.Results:The proportion of regulatory T cells (Treg)after chemotherapy in the disease control patients decreased significantly compared with that of the progressive patients (P=0.034).The difference of the proportions of Treg before and after chemotherapy affected significantly the overall survi-val (OS).The OS of the patients with decreased proportion of Treg was significantly longer than that of the patients with increased proportion of Treg,which was 23.5 and 9.4 months respectively (P<0.05). Conclusion:The patients with decreased proportion of Treg after chemotherapy had higher response rate and better survival benefit.

Objective: To investigate the potential value of 18F-fluorodeoxyglucose (FDG) PET/CT in predicting the survival of patients with primary tracheal malignant tumors. Materials and Methods: An analysis of FDG PET/CT findings in 37 primary tracheal malignant tumor patients with a median follow-up period of 43.2 months (range, 10.8–143.2 months) was performed. Cox proportional hazards regression analyses were used to assess the associations between quantitative 18F-FDG PET/CT parameters, other clinic-pathological factors, and overall survival (OS). A risk prognosis model was established according to the independent prognostic factors identified on multivariate analysis. A survival curve determined by the Kaplan-Meier method was used to assess whether the prognosis prediction model could effectively stratify patients with different risks factors. Results: The median survival time of the 37 patients with tracheal tumors was 38.0 months, with a 95% confidence interval of 10.8 to 65.2 months. The 3-year, 5-year and 10-year survival rate were 54.1%, 43.2%, and 16.2%, respectively. The metabolic tumor volume (MTV), total lesion glycolysis (TLG), maximum standardized uptake value, age, pathological type, extension categories, and lymph node stage were included in multivariate analyses. Multivariate analysis showed MTV (p = 0.011), TLG (p = 0.020), pathological type (p = 0.037), and extension categories (p = 0.038) were independent prognostic factors for OS. Additionally, assessment of the survival curve using the Kaplan-Meier method showed that our prognosis prediction model can effectively stratify patients with different risks factors (p < 0.001). Conclusion This study shows that 18F-FDG PET/CT can predict the survival of patients with primary tracheal malignant tumors. Patients with an MTV > 5.19, a TLG > 16.94 on PET/CT scans, squamous cell carcinoma, and non-E1 were more likely to have a reduced OS.