OBJECTIVE To study the distributive properties and susceptibility of yeasts to six antifungal agents. METHODS To analyze the distributive properties of 264 clinical Candida spp isolates and study the susceptibility to amphotericin B,nystatin,fluconazole,ketoconazole,miconazole and clotrimazole.The susceptibility of yeasts was tested according to the National Committee for Clinical Laboratory Standards guideline(NCCLS M27-A2). RESULTS Strains of Candida albicans were the most frequent organism isolated accounted for 62.5% of all the isolates.C.tropicalis,C.glabrata,and C.parapsilosis accounted for 20.8%,12.5%,and 1.9%,the others accounted for only 2.3%.The main infected organs were lungs,urinary tract,and digestive tract;the susceptibility tests showed strains of Candida spp to nystatin,amphotericin B,and fluconazole were more active than to the other antifungal agents.The resistance to triazole antifugal agents could be shown. CONCLUSIONS We should strengthen the diagnosis of Candida spp and strengthen the surveillance on susceptibility of clinical isolates Candida spp so as to help the doctors choose the antifungal agents reasonably.

Objective To investigate the influence of renal function on serum BNP in the diagnosis of CRF with heart failure by observing the relationship between eGFR and BNP in serum and comparing cut-off values of BNP in different eGFR levels. Methods The elderly participants were enrolled in the study, including 52 patients with heart failure, and 29 patients without heart failure and 30 healthy controls. Serum BNP level was measured by ELISA.Results The level of serum BNP increased significantly in subjects with heart failure compared with those with renal dysfunction for no-heart failure patients (P ＜ 0.05) and healthy controls. BNP level was significantly higher in CRF no-heart failure patients than in control subjects. eGFR showed negative correlation with BNP in ESRD no-heart failure patients (γ= -0. 581, P ＜ 0.01). There was no correlation between eGFR and ESRD with heart failure patients (γ= - 0.081, P ＞ 0.05). The AUC of BNP in patients (eGFR 30 ～ 60 ml) was 0. 951, when cut-off value was 1 500 ng/L,the sensitivity and specificity of BNP were 96.4% and 86. 7% respectively. The AUC of BNP in patients(eGFR ＜30 ml)was 0. 860, when cut-off value was 1 850 ng/L,the sensitivity and specificity of B NP were 66.7% and 92.9%respectively. Conclusions Heart failure was major factor result in higher levels of BNP in chronic renal failure with heart failure patients. BNP could be used as a diagnostic marker for CRF with heart failure patients.

Objective To explore the meaning of midkine (MK) levels in serum in different development stage of acute leukemia,and to explore the relationship between MK and WT1.Methods The levels of the MK in serum of 86 cases of acute leukemia and 30 cases of normal people were detected by ELISA.Real-time quantitative PCR (RQ-PCR) method was used to determine the expression of WT1 at mRNA level in 15 AML patients.Results The MK level in serum in the new diagnosed group was higher than that in the complete remission group and the normal control group [7.52 (5.44,10.55) ng/ml vs 3.52 (1.56,5.20) ng/ml vs 2.44 (1.89,3.12) ng/ml].There' s no statistical difference between midkine level in new diagnosed acute B cell leukemia (B-ALL) group and acute myeloid leukemia (AML) group [7.88 (5.78,15.78) ng/ml vs 6.25 (4.59,16.33) ng/ml].The clear correlation was found between the level of serum MK and quantities of marrow WT1 gene (r =0.529,P =0.043).Conclusions The level of MK in serum of acute leukemia patients is increased at the time of new diagnosis and decreased at complete remission.ELISA may be a way to measure the status of AL.The location of MK gene is adjacent to WT1 gene and MK' s clinical significance is similar to WT1' s,furthermore,there is a clear correlation between MK in serum and WT1 of marrow in quantities.

AIM: To look for harmfulless anti-leukemia drug with selective high performance, lethal effect of small hairpin RNA (shRNA) on VEGFR2 gene expression of tumor cell line HL60 in vitro.METHODS: The most effective VEGFR2 siRNA was designed and screened. The shRNA oligo was designed and pU6/VEGFR2 entry clone was constructed. HL60 was transfected transiently and vascular endothelial growth factor receptor 2(VEGFR2) expression was tested with MTT assay, RT-PCR and Western blotting. The expression clone was constructed and cotransfected with ViraPowerTM Packaging Mix into 293FTTM cells to produce Lentiviral vectors harboring Lenti6/shVEGFR2. The virion supernatant was added into HL60 cells and VEGFR2 gene inhibitory effect was determined. RESULTS: The inhibitory rates of VEGFR2 siRNA c were high. VEGFR2 expression in HL60 was inhibited by using pU6/VEGFR2 entry clone constructed with shRNA and pENTRTM/U6. For HL60 cells, the inhibitory rate was 84.9%. The expression of VEGFR2 mRNA and protein decreased significantly. 48 hours after transfection of pU6/shVEGFR2 entry clone and transduction of Lenti6/shVEGFR2 expression clone, the cell inhibitory rates were similar. Cell growth inhibitory rate of entry clone descended rapidly after this time point, the expression clone changed slowly, reaching the peak at 96 hours, dropped slightly, having no significance deviation. CONCLUSION: in vitro, VEGFR2 shRNA using lentiviral vector blocks VEGF/VEGFR2 self-secretion in HL60 cells, which inhibits leukemia development.

AIM:Xaf1-Saos inducible cell lines,which contain "gene switch" system were used to detect the effect of Xaf1 on tumor necrosis factor receptor(TNFR) signal pathway and to investigate the mechanism of cooperation between Xaf1 and TNF-? in inducing cell apoptosis.METHODS:Xaf1 on TNFR1 expression was measured by RT-PCR and Western blotting.The effect of NF-?B on Xaf1 induced apoptosis was detected by DNA content flow cytometry after co-transfection.DNA binding activity of NF-?B was identified by gel mobility shift assay and transcription activity of NF-?B was analyzed by luciferase assay and RT-PCR.SAPK/JNK activity was checked by SAPK/JNK assay.RESULTS:Xaf1 did not modulate TNFR1 at protein and mRNA levels.Increased NF-?B activity in cells inhibited Xaf1 induced apoptosis.Expression of Xaf1 impaired modestly TNF-? induced NF-?B DNA binding activation and transcription activation,also modestly reduced SAPK/JNK activity.CONCLUSION:Xaf1 inhibits TNFR signal pathway,partly contributing to cooperation with TNF-? to induce apoptosis.

Objective To explore the value of color doppler ultrasonography by bed side in the early diagnosis of HIE in full term neonates.Methods The changes of cerebral parenchymal and cerebral arterial blood stream parameter on 35 cases of neonates clinically diagnosed HIE of mild and moderate degree and 40 cases of normal newborns on the 24,48 and 72 hours after birth were observed by color doppler ultrasonography by bed side.Results 1.The cerebral parenchyma was even echo in normal newborns,but it was maldistributed and reinforced in mild asphyxia neonates and it was more serious in moderate degree.The echo of cerebral parenchyma in mild degree was near normal in 48 hours after birth,while the echo of cerebral parenchyma in moderate degree was still maldistributed and reinforced in 48 and 72 hours after birth.2.There was obvious changes in the cerebral arterial blood stream parameter and hemodynamics of the asphyxia newborns compared with normals.The systolic peak velocity(Vs)and end diastolic velocity(Vd)of the cerebral arteries in mild and moderate degree were obviously lower than that of control group in 24,48 hours after birth(Pa0.05).3.Resistance index(RI)of the cerebral arteries in mild and moderate degree were higher than that of control group in 24,48 hours after birth(Pa0.05).Conclusion Color doppler ultrasonography by bed side is a convenient,noninvasive method for diagnosing HIE.

The purpose of this study is to investigate the enantioselectivity of norgestrel (NG) transdermal permeation and the potential influence of linalool and lipids on the enantioselectivity. In vitro skin permeation studies of NG across the excised rat skins were performed with Valia-Chien diffusion cells, and the permeation samples were analyzed by enantioselective HPLC. The possible enantioselective permeation of NG across intact rat back skin and lipids extracted rat back skin and the influence of linalool were evaluated. The skin permeation rate of dl-NG was two times higher than that of l-NG when donor solutions (EtOH/H2O 2 : 8, v/v) containing l-NG or dl-NG. It may be mainly attributed to the solubility discrepancy between enantiomer and racemate. The enantioselective permeation of dl-NG across intact rat skin was observed when the donor solutions containing dl-linalool. The permeation flux of l-NG was 22% higher than that of d-NG. But interestingly, the enantioselective permeation of dl-NG disappeared under the same experimental condition except that the lipid extracted rat skin was used. Attenuated total reflection-fourier transform infrared spectroscopy analysis of stratum corneum showed that the wave number for asymmetric CH2 stretching vibrations of lipids treated with dl-linalool was greater than that of the control. The results indicated that the enantioselective permeation of NG may be contributed by the interaction between dl-linalool and lipids. More than half of lipids were composed of ceramides. The stereospecific interaction maybe existed among chiral enhancer (linalool), lipids (ceramides) and/or chiral drugs (NG).

Objective To probe the value of ultrasound in detection of malignant foci among multiple thyroid nodules. Methods Ultrasonograms of 83 patients with pathologically confirmed thyroid cancer combined with benign nodules were analyzed.Results Among 83 patients of thyroid carcinoma, 89 malignant and 87 benign nodules were detected with ultrasound. Significant differences of two-dimensional ultrasounic signs including low echoes, border fuzzy, micro-calcification and rear echo attenuation were found bteween malignant and benign nodules (P<0.01). There were significant differences in the grade Ⅱ, Ⅲ blood flow between malignant and benign ones (P<0.01). Of 89 malignant nodules, small thyroid carcinoma with diameter ≤1 cm accounted for 52.81% (47/89), and ultrasound diagnostic rate of all cancer nodules was 67.42% (60/89).Conclusion Ultrasounic characteristics including low echoes, border fuzzy, micro-calcification, rear echo attenuation and internal rich blood flow contribute to differentiate the thyroid microcarcinoma from multinodules.In particular, the microcarcinoma should not be ignored.

The skin ulceration syndrome of sea cucumber is a kind of desease induced by bacterium.In order to investigate the bacterium of infected sea cucumber and detect the N-acyl-homoserine lactones(AHLs) se-cretion of the bacterium,7 bacterial strains were isolated from the infected sea cucumber.These strains were identified by physiological-biochemical characteristics and 16S rDNA sequence.Results show that strain C6 belongs to Tenacibaculum,strain 4 belongs to Shewanella putrefaciens group,strain TB belongs to Vibrio,strain BP2,BP3,BP4 and BP6 belong to Pseudoalteromonas,respectively.AHLs were detected with strain Agrobacterium tumefaciens KYC55.Among these bacterial strains,strain C6,4,TB,BP3 and BP4 can se-cret AHLs,while strain BP2 and BP6 can’t.And the AHLs activity differs,from the highest to the lowest are 4,TB,BP4,BP3 and C6.

ObjectiveTo identify serum specific protein biomarkers for the early diagnosis of lymphoma patients by using surface enhanced laser desorption/ionization time of flight mass spectrometry (SELDI-TOF-MS)protein chip array.MethodsSELDI-TOF-MS technique and weak cation exchanger (WCX2)protein chip were used to detect the serum proteomic patterns of 96 patients with lymphoma (86 patients with non-Hodgkin lymphoma and 10 patients with Hodgkin lymphoma)and 30 normal control.The serum level of LDH were measured by biochemical methods,the serum level of β2-MG and CA125 were detected by enzyme-linked immunosorbent assay (ELISA).ResultsFour protein peaks pattern mass/charge ratio (M/Z) 6880,8564,8692 and 13751 were decreased in lymphoma patients,and they could be used to distinguish lymphoma from healthy people and other malignant tumors.Their sensitivity and specificity were 82.29 ％ and 80.00 ％,78.13 ％ and 73.30 ％,75.00 ％ and 80.00 ％,71.88 ％ and 83.30 ％ respectively in lymphoma patients.When it was used to early diagnosing lymphoma in stage Ⅰ or Ⅱ,the sensitivity of four protein peaks pattern (M/Z 6880,8564,8692 and 13 751) were 77.55 ％,71.43 ％,71.43 ％ and 67.35 ％respectively and the specificity of all four protein peaks pattern were 100.00 ％.Four protein peaks pattern sensitivity were higher than LDH,CA125 and β2-MG respectively.ConclusionApplication of SELDI-TOF-MS can be of great potential for the early diagnosis of lymphoma patients.