AIM: To investigate whether and how N, N-dimethylsphingosine (DMS) plays a role in modulating the adhesion of monocytes to vascular endothelial cells, and identify whether human umbilical vein endothelial cell line EA.hy926 take place of the vascular endothelial cells.METHODS: Adhesion ratio was measured by flow cytometry, and immunohistochemistry was used to detect the expression of ICAM-1 and P-selectin in HUVEC: EA.hy926 cells after the effect of DMS. RESULTS: DMS inhibited the adhesion of monocytes to HUVEC: EA.hy926 cells in a time-dependent and concentration-dependent manner by reducing the expression of ICAM-1 and P-selectin. CONCLUSIONS: DMS reduced adhesion molecule expression in vascular endothelial cells. DMS may be an important contributor to reduce adhesion ratio, suggesting that DMS plays a negative role in proinflammatory and immune functions of the modified vascular endothelial cells during atherosclerosis and restenosis.

Objective To investigate the relationship between the activity of Na +/K +pump on RBC membrane and abnormal potassium transportation through cellular membrane in hypokalemic patients Methods Activity of RBC ATPase in 17 cases of hypokalemic paralysis patients and 16 cases of healthy controls was tested Results Activity of RBC ATPase was statistically higher in hypokalemic periodic paralysis (HOPP)group[( 25 6?3 5) ?mol?(10 7RBC) 1 ?h 1 ], and lower in nephridium oxidosis group [(1 9?0 8) ?mol?(10 7RBC) 1 ?h 1 ]than in control group[(6 2?0 9) ?mol?(10 7 RBC) 1 ?h 1 , P

Objective To study the relations between CD23 expression on peripheral blood B lymphocytes and IgE in asthmatic children and their pathogenesis. Methods CD19 +, CD23 +, CD19 +/CD23 + expressions of peripheral blood lymphocytes were measured by flow cytometry and serum total IgE level by chemiluminescence immunoassay in 22 children with classical asthma, 18 cough variant asthma (CVA), 20 with asthmatiod bronchitis ,20 with pneumonia and 25 healthy controls. The correlations between CD23 expression on peripheral blood B lymphocytes and IgE in serum were analysed. The 20 children with asthmatiod bronchitis were divided into two groups by the scores whether it reached that for diagnosing asthma after ten months: Group Ⅰ (10 cases) and Group Ⅱ (10 cases). Results CD19 +, CD23 +, CD19 +/CD23 + expressions on lymphcytes were significantly elevated in peripherial blood and serum IgE from children with classical asthma [(13.56?5.87)%,(26.56?7.61)%,(12 86?5.01)%, (12.67?7.56) ?mol/L respectively] and CVA [(13.10?5.15)% ,(24.66?8.15)% , (12 10?4.97)%, (9.45?4.16) ?mol/L respectively] as compared with those in the children with pneumonia and healthy controls [(7.95?3.65)%, (14.92?3.02)% ,(6.82?2.95)% , (3.10?1.68) ?mol/L respectively] ( P

Objective To investigate the differences and harmonization of immunoassay systems in detecting CA19-9 and to assess the possibility of mutual recognition in different laboratories.Methods Data were collected and analyzed from External Quality Assessments (EQA) of NCCL(Lots:200811-201215) and ZJCCL(Lots:080309-120211).120 fresh serum with different concentrations of CA19-9 were collected.The CA19-9 results of healthy people were also collected from September 2010 to March 2012.Four kinds of stable immunoassay systems were involved in our research,including Abbott Architect i2000,Beckman UniCel DxI 800,Roche E170 and Siemens ADVIA Centaur XP.The differences among four system groups were calculated with the EQA data.The fresh serum comparisons were also performed following the guideline of CLSI EP9-A2 The 95％ confidence interval of each immunoassay system was calculated.Comparisons were made by scatter diagrams and weighted regression.Results Both EQA of NCCL and ZJCCL showed better correlation coefficients and larger bias (bw ranged from 1.340 to 4.683) than in fresh serum comparisons.Although the correlation coefficients were all unsatisfactory,the bw were all close to 1 in fresh serum comparisons.When the recommended serum concentration of 27 U/ml was used,the biases were Abbott-Roche-6.41％,Beckman-Roche-5.07％,Siemens-Roche 13.15％,Beckman-Abbott 2.46％,Siemens-Abbott 22.52％ and Siemens-Beckman 39.66％,respectively.Differences of 95％ confidence intervals were statistically significant among parts of the immunoassay systems.Conclusions Only in the lower concentration can CA19-9 results be mutual recognized among four different immunoassay systems,there will be larger differences and risks in the higher concentration.

Objective To observe the inhibition of amikacin in vitro on platelet aggregation and blood coagulation tests, and to study its effects on hemostasis and the related mechanisms.Methods Plateletrich plasma and platelet-poor plasma from donors were mixed with different concentration of amikacin, which was divided into four groups:0 mg/L, 30 mg/L, 91 mg/L and 910 mg/L group.The maximial ratio of platelet aggregation induced by ADP were measured with Platelet Aggregation Analyzer.The expression levels of P-selectin, GP Ⅱ b/Ⅲ a and Fg-R were determined with Flow Cytometer.The PT, APTT, TT and Fg of platelet-poor plasma were detected with Blood Coagulation Analyzer. The four concentration of amikacin mentioned above and two anticoagulants (62.5 U/ml of sodium heparin and 109 mmol/L of sodium citrate)were interacted with fresh whole blood, in which the blood CT and plasma Ca2+ were detected. Blood samples were collected from 10 patients with acute lower respiratory tract infection before and 30 minutes after routine amikcin treatment respectively.The maximial ratio of platelet aggregation, the expression levels of P-selectin, GPⅡ b/Ⅲa and Fg-R induced by ADP were measured; while PT, APTT, CT and plasma Ca2+ were determined.Results At 30 mg/L of amikacin group, the maximal ratios of platelet aggregation (65.8±3.9)%, the expression levels of P-selectin (9.2 ± 1.0)% and Fg-R (12.6 ± 1.7)% were statistically lower than those [(88.0 ±4.6%, (16.1 ± 1.3)% and (31.0 ±2.5)%]at 0 mg/L of amikacin group ( t = 9.442,8.432,9.993,P ＜ 0.01 ).At 30 mg/L of amikacin group, the APTT (80.5 ±6.8) s and CT ( 857 ± 66) s were significantly higher than those [(33.0 ± 3.6) s and (447 ± 35 ) s] at 0 mg/L of amikacin group ( t = 11.312, 13.211, P ＜ 0.01 ). There was a negative correlation between amikacin concentration and maximial ratio of platelet aggregation ( r = - 0.832, P ＜ 0.05 ), but a positive correlation between amikacin concentration and inhibitory rates of platelet aggregation ( r = 0.939, P ＜0.05) was observed, as well as APTT (r ＞0.870, P＜0.05).At 30 mg/L, 91 mg/L, and 910 mg/L of amikacin groups, the P-selectin and Fg-R expression were remarkably inhibited with a dose-dependent manner, the CT was notably enhanced [Fwithin subjects =21.44, 26.24, ( ＞29.81 ), P ＜0.01].At 0 mg/L,30 mg/L, 91 mg/L and 910 mg/L of amikacin groups, the PT values were ( 14.7 ± 1.9) s, ( 15.2 ± 1.7) s,(15.6±1.5) s and (22.1 ±2.1) s, respectively (F=8.21,P＜0.05), but there was no markeddifference for the levels of GP Ⅱ b/Ⅲ a, TT, Fg and plasma Ca2+ among the four groups ( P ＞ 0.05 ).After 30 minutes of amikacin treatment, the maximial ratio of platelet aggregation (51.6 ± 10.1)%, the expression levels of P-selectin (6.8 ± 1.8) % and Fg-R ( 20.1 ± 5.8 ) % were significantly lower than those [(66.8 ± 11.4)%, ( 10.9 ±3.1 )% and (28.5 ±7.4)%] before amikacin treatment, but APTT (49.8 ±5.9) s and CT (660 ±59) s were remarkably higher than those [(26.9 ±3.8) s and (410 ±45) s] before amikacin treatment, respectively ( t = 5.456,8.875,7.423,10.012,11.322, P ＜ 0.01 ), while the GP Ⅱ b/Ⅲ a expression, PT and Ca2+ concentration had no significant changes ( P ＞ 0.05).Conclusions There are inhibitory effects of amikacin on platelet aggregation mainly through the inhibition of both fibrinogen receptor activation and secretion reaction of activated platelet. Amikacin may also inhibit pathway of coagulation system factor to prevent blood coagulation.Therefore, risk of hemorrhage may be investigated in the patients with amikacin for anti-infection treatment.

Objective To investigate the effects of ZGDHu-1 on T lymphocytes activation in vitro to elucidate its immunosuppressive effects. Methods Lymphocytes isolated from healthy persons were stim-ulated with phytohanmagglutinin(PHA) and different experimental groups were set by cocultured for 24 h, 48 h with ZGDHu-1 or with ZGDHu-1 and Cyclosporin A(CSA). To assess the proliferation and apoptosis of T lymphocytes, we detected CD3~+ CD69~+ , CD3~+ CD25~+ , CD4~+ CD25~+ , CD8~+ CD25~+ and CD3~+ Fas~+, CD4~+ Fas~+ , CD8~+ Fas~+ with flow cytometry. The early apoptosis rate of lymphocytes was analyzed with flow cytometry. Culture supernatant IL-2 and TGF-β1 were detected with ELISA. Results ZGDHu-1 decreased PHA activative CD3~+ CD69~+, CD3~+ CD25~+, CD4~+ CD25~+ and CD3~+ Fas~+, CD4~+ Fas~+, Annexin V~+/ PI~- and inhibited IL-2 secretion and promoted TGF-β1 secretion respectively. ZGDHu-1 has synergistic effect with CSA to be more obvious. Conclusion ZGDHu-1 can inhibit T lymphocytes activation and de-creased apeptosis of T lymphocytes. ZGDHu-1 has synergistic effect with CSA to be obvious.

Objective To explore the inhibition effects of CD4+CD25+/highCD127low on the EBV immortalized cells(CD23+)in children with infectious mononucleosis (IM). Methods The expression of CD3+, CD3+ CD4+ , CD3+ CD8+ , CD25+/highCD127low and CD19+ , CD19+ CD23+ were analyzed by flow cytometry in 23 children at the acute stage of IM and 10 children recovering from IM in comparison with the ones of 20 healthy controls. Results Compared with the following results in controls, CD3+, CD3+ CD8+ were significantly increased and CD3+ CD4+, CD4+/CD8+ ratio, CD4+ CD25+/highCD127low, CD19+, CD19+ CD23+ were significantly reduced in the IM patients. Compared with the ones in the IM pa-tients recovering from IM, CD3+ , CD3+ CD8+ were significantly increased and CD3+ CD4+, CD4+/CD8+ ratio, CD4+ CD25+/highCD127low, CD19+ , CD19+ CD23+ were significantly reduced in the IM patients at the acute stage. There was no obvious difference in all the marks except CD4+/CD8+ratio, CD8+ CD28+ , CD19+ , CD19+ CD23+ ratio between the children recovering from IM and the normal controls. There is a positive correlation between CD3+ and CD19+, CD19+ CD23+ during acute phase. There is a negative correlation between CD4+ CD25+/highCD127low and CD19+ , CD19+ CD23+ during acute phase. Conclusion The results indicate that the decreasing of CD4+ CD25+/highCD127low may play an important role in elimina-ting EBV immortalized cells.

Objective To establish a flow cytometry-based drug susceptibility test for the rapid de-tection of antifungal susceptibility or resistance of Candida isolates.Methods The gate selection and opti-mal experimental conditions of flow cytometry-based drug susceptibility test were determined by using Candi-da albicans strain ATCC90029 as the test strain and propidium iodide ( PI) as the fluorescent dye .The es-tablished flow cytometry-based drug susceptibility test was used to detect the susceptibility or resistance to fluconazole or voriconazole of 110 isolates belonging to Candida species, and the obtained results were com-pared with those by using typical M 27-A3 constant dilution method .Results The killed and viable Candida albicans ATCC90029 strains were clearly divided into two groups on the figure of SS /log (FL3) by regulating voltages.There was a high correlation between the results of susceptibility test and the proportions of killed and viable fungi in mixture (r=0.999).The flow cytometry-based drug susceptibility test could provide the results within 30 min and its optimal concentration of fungal suspension , time of drug-fungus incubation , dyeing method and time were 1.0×106/ml, 3 h incubation and sodium deoxycholate-pretreated plus PI dye-ing for 5 min, respectively .The total coincident rates between the established test and the constant dilution method were 98.2%and 87.3%in the detection of drug susceptibility of 110 fungal isolates to fluconazole and voriconazole .Conclusion The flow cytometry-based drug susceptibility test shows advantages of rapidi-ty, accuracy and high sensitivity compared with the constant dilution method .It has a great potential for clin-ical application .

We isolated and identified the bacterial pathogen in a pyogenic balanoposthitis patient and investigated the drug resistance and its mechanism of the isolate.Urethral secretions and balanus pustule liquids were collected for microscopic examination after Gram-staining and detection of mycoplasma using Mycoplasma IST 2 kit.The two samples were inoculated on Columbia blood plate,N.gonorrhoeae selective plate and chromID Candida plate for isolation.The obtained colonies were identified by VITEK 2-compact automatic bacterial detection and analysis system.Moreover,PCR was performed to detect 16S rRNA gene of N.gonorrhoeae in the samples and colonies.KB method was applied for detecting susceptibility of five common antibiotics against the isolate.The β-lactamase and extended spectrum β-lactamase confirmatory tests were used to investigate the enzyme production of the isolate as well as drug resistance-associated tetM,TEM,mefA and ermF genes in the isolate were detected by PCR.Results showed that all the clinic samples showed negative for mycoplasma.All the isolating cultivation results of urethral secretions were negative while the balanus pustule liquids provided positive isolating cultivation in the blood and selective plates.The VITEK 2-compact system and 16S rRNA-PCR revealed that the isolated strain belongs to N.gonorrhoeae.The isolate can produce β-lactamases and resist to penicillin G,ciprofloxacin and tetracycline.The tetM,TEM,mefA and ermF genes could be found in the isolate's genome.The patient's balanoposthitis is caused by infection of N.gonorrhoeae.The multidrug resistance of Neisseria gonorrhoeae isolate is closely associated with its carried resistant genes.

0.05).There was a positive corrlation on CD40~+ and CD40L~+ in chronic B hepatitis patients.There was a positive corrlation on CD8~+/CD28~+ and CD40~+、CD40L~+ in chronic B hepatitis patients.There were no remarkable corrlation on CD8~+/CD28~- and CD40~+、CD40L~+ in chronic B hepatitis patients.Conclusion:The costimulation molecules CD40~+、CD40L~+ and CD8~+/CD28~+ are lower,while CD8~+/CD28~- are higher in chronic B hepatitis patients than in the health.To test the expression of CD40~+、CD40L~+ and CD8~+/CD28~+ on peripheral blood of the chronic B hepatitis could help to evaluate patients's celluar immunity and guide clinical treatment.