Objective To investigate the effects of UVB irradiation and calciu m at different concentrations on the expression of pemphigus antigens by culture d human keratinocytes. Methods Human keratinocyte cultures were treated with eit her 2 mmol/L calcium added to the serum free media, or exposure to UVB irradiat ion. Immunofluorescence staining was performed with sera from patients with pemp higus vulgaris (PV) or pemphigus foliaceus (PF) as the first antigen. Extracts f rom both epidermis and keratinocytes were run on SDS-PAGE according to Laemmli ′s method and transferred to nitrocellulose membrane for immunoblot with PV and PF sera. Results Specific staining with PV sera was always detectable on kerati nocyte culture by immunofluorescence assay with or without high concentrations o f calcium while PF antigen was detected on stratified cells only. However, expos ure to UVB irradiation could not evoke keratinocyte culture express PF antigen. The reactivities were found at 160 kd band with PF sera while at both 160 kd and 130 kd bands with PV sera. Conclusions Monolayer or stratified keratinocytes ca n produce PV antigen, by increasing the concentration of calcium in the culture media, the human cultured kera tinocytes can be induced to stratify and express PFA. Human keratinocytes can not express PF antigen after exposure to UVB in intro.

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 analyze the sperm DNA fragmentation in different degrees of varicocele (VC) infertility patients after high ligation of the spermatic vein. Methods There were 57 patients with different degrees of VC infertility patients. 27 patients had been diagnosed with level two VC, and 30 patients had been diagnosed with level three VC. All patients' sperm DNA fragmentation index (DFI) were detected and calculated at preoperative one month and postoperative three months. Results 57 VC infertility patients were all suffered from laparoscopic high ligation of the spermatic vein on both sides. There was no statistically difference on preoperative sperm DFI between level two and level three patients (P > 0.05). All patients' sperm DFI were decreased after laparoscopic (P < 0.05), and level two VC infertility patients had significant greater progress than level three patients (P < 0.05). Conclusion Laparoscopic high ligation of the spermatic vein can effectively improve VC infertility patients sperm DFI. It would be better for level two VC infertility patients.

Objective To investigate the sensitivity of dihydroartemisinin against different developmental stages of Schistosoma japonicum, so as to understand the effect of dihydroarteminisin against on S. japonicum. Methods Mice were infected with 40 S. japonicum cercariae on the abdomen. Dihydroartemisinin was given intragastrically at different developmental stages of S. japonicum , and the mice were sacrificed 50 days post-infection, the adult worms were collected, and the worm reduction rates and female worm reduction rates were calculated. ① On the 2nd h, 3rd, 5th, 7th, 10th, 14th, 18th, 21st, 28th, 35th day post-infection, the mice were administered intragastrically with dihydroartemisinin at a single dose of 300 mg/kg, and the effect of dihydroarteminisin on different developmental stages of S. japonicum were observed. ② The mice were administered with different doses of dihydroartemisinin on the 7th or 35th day post-infection, and the dose-effect was explored. ③ The mice were administered on the 7th day post-infection and re-administered on the 35th day post-infection, respectively with different doses of dihydroartemisinin, and the effect was evaluated. Results The dihydroartemisinin at a single dose of 300 mg/kg had obvious effect on 7-day schistosomula and 35-day adult worms, with 64.81% and 60.47% of worm reduction rates and 73. 81% and 90.48% of female worm reduction rates, respectively. When the mice on the 7th day post-infection were administered with 200, 300, 400 and 600 mg/kg dihydroartemisinin, the worm reduction rates were 46.84% , 60.63% , 59.55% and 60. 21% , respectively, and the female reduction rates were 59.73% , 72.29% , 72.58% and 76.61 % , respectively. While in the mice on the 35th day post-infection, the corresponding rates were 47. 23% , 62.33% , 76.31% and 83.63% , respectively, and 59. 73% , 89. 36% , 89.65% and 93.96% , respectively. When the mice were treated twice with dihydroartemisinin on the 7th day and 35th day post-infection, the worm reduction rates were 58. 16% , 82.66% ,83.42% and 83.79% , respectively, and the female worm reduction rates were 68.69% , 90.43% , 93.74% and 94.63% , respectively. Conclusions Dihydroartemisinin has effect against S. japonicum, and the 7-day schistosomulum and 35-day adult worm are sensitive to the drug.

Objective To investigate the resistance of carbapenems,the production of metallobeta-lactamases(MBLs)and homology analysis of Psendomonas aeruginosa in China.Methods A total of 654 Pseudomonas aeruginosa isolates were collected from 28 hospitals in 16 difierent regions during the period from July 2006 to July 2007 as part of the Nosocomial Pathogen Resistance Surveillance(NPRS).MICs of imipenem and meropenem were determined by agar dilutiom PFGE was used to analyze the molecular epidemiology of carbapenem-resistant strains.MBLs were detected by Etest and PCR among carbapenem-non-sensitive strains.The transcription levels of MBLs were determined by real-time reverse transcriptase(RT)-PCR.Results Of the 645 Pseudomonas aeruginosa strains,245(37.5%)were resistant to imipenem and 183(28.0%)were resistant to meropenem.A total of 275 carbapenemnonsusceptible strains and 259 carbapenem-resistant strains were chosen for further study. Among the carbapenem-nonsusecptible strains,8.73%(24/275)of the strains carrried MBLs genes hy PCR,but only 6.55%(18/275)of the strains were detected as MBls-producers bv MBL Etest. The MBLs relative expression levels of the 6 MBL genotype-positive but phenotype-negative strains were significantly lower than those of the other 18 MBL genotype-and phenotype-positive strains(P＜0.05).The 259 carbapenemresistant Pseudomonas aeruginosa strains were divided into 89 clones by PFGE.The 24 MBL genotypepositive strains collected from 6 cities were divided into 9 clones by PFGE.Conclusions The carbapenems resistance of Pseudomonas aeruginosa is severe in China.Clonal dissemination of Pseudomonas aeruginosa has not been detected throughout China.MBLs are not the major carbapenems resistance mechanism in Pseudomonas aeruginosa.

Objective To investigate the resistance of Pseudomonas aeruginosa and genotyping of the main β-lactamases in China. Methods A total of 645 Pseudomonas aeruginosa isolates were collected from 28 hospitals in 16 cities in China from July 2006 to July 2007. The susceptibilities to 11 kinds of antimicrobial agents were detected by agar dilution or Kirby-Bauer disk diffusion method. The genotypes of β-lactamases including TEM, SHV, CTX-M and OXA of all the strains were detected by polymerase chain reaction (PCR) and sequence analysis. Results The resistance rates of 645 Pseudomonas aeruginosa isolates to antimicrobial agents were high, except those to amikacin and meropenem were lower than 30 %. Two hundred and seventy-five (42. 64 % ) strains were carbapenem and (or) meropenem-nonsusceptible Pseudomonas aeruginosa. Three hundred and sixty-eight (57.05 %) strains were multidrug-resistant Pseudomonas aeruginosa and 20 (3. 10%) strains were pandrug-resistant. The genotyping results of β-lactamases were as follows: 51 stains produced OXA-10 group β-lactamases, 37 were CARB type, 36 were TEM, 35 were PER, 11 were CTX-M, 9 were VEB, 5 were SHV, 24 were metallo-β-lactamases positive and 1 was GES. None of genotypes of plasmidmediated AmpC enzyme and other carbapenemases were detected. CTX-M-13, CTX-M-14,CTX-M-15, CTX-M-3 of extended spetrum β-lactamese were detected in Pseudomonas aeruginosa.Conclusions The situation of Pseudomonas aeruginosa resistances is severe in China. OXA-10 and PSE-1 are the most common genotypes of β-lactamases. The β-lactamases genotyping is different between carbapenem-nonsusceptible and carbapenem-susceptible strains.

Objective To research the homology of 18S small subunit ribosomal RNA(18S-rRNA) gene about Chinese Mainland and Philippine strains of Schistosoma japonicum,and Schistosoma mansoni,and the possibility to establish the PCR assay based on the gene for detecting the cercaria in a low density level. Methods The genomic DNAs of Chinese Mainland and Philippine strains of S.japonicum,and S.mansoni were extracted. The PCR assay was used to detect the identical target DNA elements in the above genome team and the homology of their genes was compared. The single cercaria was respectively treated with the method of heating in boiling water,the method of treating with ammonia and the method of treating with NaOH,HCl and ethanol,and the single treated cercaria and the single cercaria without treating were used as the templates to amplify the target DNA by using the PCR assay,and the detection rates of the PCR assay to detect the single cercaria treated with the different methods were calculated and compared. Results With the genomic DNAs of Chinese Mainland and Philippine strains of S.japonicum and S.mansoni as the templates,the target DNA element of which sequence length was 469 bp was all amplified by PCR. The target DNA was all amplified by PCR to the single cercaria treated with ammonia and the method of treating with NaOH,HCl and ethanol. However,only 50 percent of specimens of the single cercaria without treating and the single cercaria treated with the method of heating in boiling water were amplified to the target DNA by PCR. Conclusions The 18S-rRNA gene has the general homology among the species and strains of Schistosoma. The sensitivity of the PCR assay to detect the low density cercaria treated with ammonia or the method of treating with NaOH,HCl and ethanol is higher than that of the single cercaria without treating or treated with the method of heating in boiling water.

Purpose To assess the clinical and pathological features of small gastrointestinal stromal tumours (sGIST).Methods To reevaluated the clinical,histological and immunohistochemical parameters of 21 sGISTs.The standard immunohistochemical panel antibodies were studied on the tumor sections.All data were compared with clinical sGIST.Results There were a total of 7 females and 14 males of sGISTs.The median age was 63 years old.The tumors were predominantly located in the stomach showing a spindle cell morphology and the tumor sizes ranged from 0.5 cm to 1.5 cm.9 sGISTs combined with malignant tumors,which were gastric cancer have been incidentally detected during surgery.As the lesions were small in size,with infrequent bleeding,necrosis,mucosal invasion,ulceration and less mitotic index,sGISTs reoccurred less compared with clinical sGIST.p53,Ki-67 labeling index and microvascular density (MVD) in sGIST were significantly lower than clinical sGIST (P ＜ 0.05).Conclusion sGIST may occure with digestive tract cancer synchronously.p53,Ki-67 labeling index and MVD were lower than clinical GIST,which means better prognosis.

AIM: To investigate the effects of angiotensin-Ⅱ (Ang-Ⅱ) on the proliferation and collagen synthesis in rat cardiac fibroblasts and the expression of Ang-Ⅱ receptor on fibroblasts. METHODS: [3H]-TdR and [3H]-prolin at different concentrations were incubated with the confluent fibroblasts of newborn rats stimulated by Ang-Ⅱ. Receptor of Ang-Ⅱ on fibroblasts and intracellular free calcium ions were examined by autoradiograms and fluorescence technique. RESULTS: In the presence of Ang-Ⅱstimulation, the increase in incorporation of [3H]-TdR and [3H]-prolin was much higher than that of control in a dose-dependent manner. Autoradiograms showed that Ang-Ⅱ was uniformly distributed over the membrane of the fibroblasts. The silver grains on fibroblasts were obviously decreased after adding unlabeled Ang-Ⅱ by the autoradiograms. The concentration of free calcium ions was increased in fibroblast. CONCLUSION: These findings suggest that Ang-Ⅱ promotes fibroblast proliferation and the syntheses of collagen protein, in which Ang-Ⅱ binds to the membrane receptor of fibroblasts to activate the signal system in cells, such as intracellular free calcium ions. [

Objective To investigate the etiology of hand,foot and mouth disease (HFMD) and to analyze the genetic characteristics of three prevalent enteroviruses in Qingdao in 2015. Methods City-wide surveillance data in 2015 were used to analyze the epidemiological characteristics of HFMD in Qingdao. RNA samples extracted from throat swab of HFMD cases were examined for general enteroviruses (EVs) such as enterovirus 71(EV71), coxsackievirus A16 (CA16) and CA6 by real-time RT-PCR. Full-length VP1 genes of EV-positive specimens were amplified and sequenced. Sequencing results were phylogenetically analyzed using MEGA 7.0 software package. Results A total of 804 patients with HFMD were identified from 1 176 patients in 2015. CA6 (41.4%),EV71 (31.6%) and CA16 (15.3%) were the predominant EVs causing HFMD. Children 5 years of age and under accounted for 80.3% of the 804 HFMD cases. CA6 was responsible for 48.9% of HFMD cases in children under 3 years old. The epidemic subtypes of EV71 and CA16 in Qingdao were C4a and B1b,respectively. Twenty-eight randomly selected CA6 strains were all classified into type A genome. Conclusion CA6, EV71 and CA16 were the prevalent pathogens causing HFMD in Qingdao in 2015. CA6 became the most predominant pathogen,mainly targeting children under 3 years old. C4a remained the prevailing subtype of EV71,while different from the co-prevalence of B1a and B1b subtypes in the past,B1b became the predominant subtype of CA16. CA6 strains circulating in Qingdao in 2015 mainly belonged to type A genome and evolved into multiple smaller branches. However, CA6 strains isolated in Qingdao in 2015 and 2013 located in different branches.