OBJECTIVE To establish a simpler method for screening bacterial metallo-?-lactamase(MBL) by comparing Hodge test and double-disk synergy tests(DDSTs),and using 5 inhibitors such as sodium citrate,EDTANa2,EDTAK2,sodium dimercaptopropane sulfonate and mercaptoethanol.METHODS In Hodge test,5 inhibitors of different concentration were added to the imipenem(IMP) disks,the sizes of the inhibition zones were compared.Disk containing IMP and disk containing a metallo-?-lactamase inhibitor were used in DDSTs.The optimal inhibitors concentration and edge-to-edge distance between the two disks were selected.The performances of the Hodge test and the DDSTs were also compared.RESULTS Among the metallo-?-lactamase inhibitors used in this study,1∶15 diluted mercaptoethanol gave the most reproducible and the clearest results when a filter disk containing mercaptoethanol was placed near the IMP disk with the edge distance of 3 mm.CONCLUSIONS Mercaptoethanol-IMP DDSTs are simple,convenient and sensitive methods for screening MBL.

Objective To investigate the clinical characteristics of the tympanic membrane atelectasis and treatment methods ,and to provide a reference value for future clinical diagnosis and treatment .Methods A retro‐spective analysis of 86 patients(104 ears) with tympanic membrane atelectasis treated in our hospital from June 2011 to August 2013 .Disease severity was classified according to the erasmus classification of atelectasis by Sade ,and pre- and post -operative air -bone gaps (ABG) were compared .Results There was no statistical difference of mastoid gasification on CT scan between mild and sever tympanic membrane atelectasis (P>0 .05) .While the sta‐tistical difference was found in two groups of whether there were soft tissues in middle ear and mastoid cavity on CT scan(P<0 .05) .There was an improvement in the average ABG for all stages .Conclusion This study demonstrated that surgical intervention had a favorable effect on hearing level across all stages .The treatment of atelectatic ears should be taken and individualized .

Objective To evaluate the role of complement receptor 3 (CR3) on murine macrophages in the recognition of Penicillium marneffei.Methods RAW264.7 murine macrophage cells were cultured in vitro,and divided into four groups to be cocultured with inactivated and live Penicillium mameffei yeast cells as well as inactivated and live Penicillium marneffei conidia respectively at 37 ℃ in 5％ CO2 for one hour.The RAW264.7 cells incubated with phosphate-buffered saline (PBS) served as the blank control group.Then,reverse transcription-PCR was conducted to detect CR3 mRNA expression,Western blot to measure CR3 protein expression,flow cytometry to determine phagocytosis rate,enzyme-linked immunosorbent assay (ELISA) to quantify cytokine levels in culture supernatant.Some RAW264.7 macrophages were transfected with a specific siRNA targeting CR3 gene and cocultured with inactivated Penicillium marneffei conidia,subsequently,phagocytosis rate and supematant cytokine levels were determined.Data were processed by the SPSS 16.0 software,and one-way analysis of variance (ANOVA) was conducted for inter-group comparisons of these parameters.Results No significant differences were observed in the mRNA or protein expressions of CR3 among the four groups of RAW264.7 cells cocuhured with different forms of Penicillium marneffei (both P ＞ 0.05).The phagocytosis rate was 95.14％,89.56％,91.03％ and 90.78％ in RAW264.7 cells cocultured with inactivated conidia and yeast cells,as well as live conidia and yeast cells of Penicillium marneffei,respectively (P ＞ 0.05).The levels of interleukin (IL)-2,interferon (IFN)-γ,IL-4 and IL-10 in culture supernatant were increased at different degrees after one-hour coculture in the four coculture groups compared with the blank control group,but no statistical difference was noted among the four coculture groups in the supernatant levels of these cytokines (all P ＞ 0.05).After coculture with inactivated Penicillium marneffei conidia,the siRNA-transfected RAW264.7 cells showed a statistical decrease in phagocytosis rate (10.89％ vs.92.78％,P ＜ 0.05) and supernatant levels of IL-2,IFN-γ IL-4 and IL-10 compared with untransfected RAW264.7 cells.Conclusions In early stage of innate immunity,CR3 on macrophages may be one of the pattern recognition receptors participating in the recognition and mediation of phagocytosis of Penicillium marneffei.It's possible that both Thl-and Th2-type cytokines,such as IL-2,IFN-γ,IL-4 and IL-10,are involved in the immune response of macrophages against Penicillium marneffei.

Objective:To investigate the protective effect of Yiqi Huazhi recipe Quqian granules on rat renal tubular cell apoptosis induced by lead poisoning. Methods:Totally 60 Wistar rats were divided into 2 groups, 12 in the control group and the others in the model group. Chronic lead poisoning model was made by drinking 0. 02% lead acetate water for 60 days. Then the lead poisoning rats were randomly divided into four groups, high-dose Quqian granules group (3. 0 g·kg-1·d-1), low-dose Quqian granules group (0. 6 g·kg-1 ·d-1 ) , positive control group ( calcium disodium edentate plus procaine, im, 50 mg·kg-1 ·d-1 ) and model group. Seven treatment courses were carried out in the first three groups with every 4-d as one course and 4-d withdrawal period between every two courses. After 60 days, the change of lead in blood and kidney was observed by atomic absorption spectrometry,the apoptosis of kidney tissues was studied by TUNEL, the expression of Bcl-2 protein was detected by immunohistochemical methods and the expression of p53 was studied by Western Blotting. Results:Compared with the control group, the body weight, hemoglobin and the expression of Bcl-2 in the model group were decreased significantly(P<0.01)those in, and Pb in blood(0.990 ±0.443)μg·ml-1, Pb in kidney(51.33 ± 5. 16)μg·ml-1 , the apoptosis of tubular epithelial cell(4. 148 ± 0. 414) and the expression of p53 protein (1. 868 ± 0. 139) were significantly higher (P<0. 05). Compared with the those in model group, the body weight, hemoglobin and Bcl-2 in high-dose group were increased significantly(P<0.01), and the blood lead level (0.082 ±0.015)μg·ml-1, the kidney lead level (6.38 ±0.97)μg ·ml-1 , the apoptosis of tubular epithelial cell(1. 412 ± 0. 109) and p53 protein expression(1. 164 ± 0. 172) were significantly lower (P<0. 05). Conclusion:Lead may induce high expression of p53,low expression of Bcl-2 and promote the apoptosis of renal tubular epithelial cells. It is proven that Yiqi Huazhi recipe Quqian granules can inhibit the expression increase of p53 and the expression de-crease of Bcl-2 resulting in the reduction of the renal tubular apoptosis to allivate the renal injury caused by lead.

This paper introduces the method of mixed programming with Labview and Matlab, and applies this method in a pulse wave pre-processing and feature detecting system. The method has been proved suitable, efficient and accurate, which has provided a new kind of approach for biomedical signal analysis.
Heart Rate ; Signal Processing, Computer-Assisted ; instrumentation ; Software

Objective To generate of human induced pluripotent stem cells from umbilical cord matrix cells(UMC).Methods Sox2 and Klf4 and Oct4 and c-Myc were transfected into UMC cells with retrovirus,and thcn UMC cells was reprogrammed to iPS cells.Gene expression was confirmed with RT -PCR and the integration was confirmed with cell karyotype.iPS cells were further validatcd with cell alkaline phosphatase detection and immunofluorescence staining,differentiating into teratomas in vivo and embryoid bodies in vitro.Results iPS cells were similar to embryonic stem cells (ES).The expression of Nanog,Oct4,Rex1 and Sox2 in iPS cells were higher than UMC cells,and Sox2,Klf4,Oct4,c-Myc silenced in iPS cells.Exogenous genes were inserted into the nucleus of iPS cells,which was confirmed by 1％ agarose gel electrophoresis.iPS ccll karyotype was normal,alkaline phosphatase activity increased,ES cell-specific proteins,including SSEA3,SSEA4,TRA-1-60,TRA-1-81 and Nanog,were expressed.iPS cells were differentiated into a teratoma in vivo and embryoid bodies in vitro.Conclusions iPS cells were similar to ES cells,which had pluripotency.

Objective To investigate the risk factors and outcomes of acute kidney injury (AKI) in patients after intra--coronary stent implantation.Methods A retrospective and case control study was done with data analysis in 325 patients who underwent intra-coronary stent implantation from January 2010 to March 2011.The patients were divided into two groups as per the criteria of AKI identified on the 7th day after implantation of stent.The variables to be studied included:(1) age,gender,hypertension,diabetes,cerebrovascular disease,left ventricular insufficiency,peripheral angiopathy,creatinine,urea nitrogen,estimated glomerular filtration rate,hyperuricemia,proteinuria,emergency operation,hydration,and medication (ACEI/ARB,statins) before operation; (2) dose of contrast media,operation time,hypotension during intra-operative period; and (3) postoperative:hypotension.The variables were analyzed with the process of One-way ANOVA and multivariate Logistical regression analysis.Consequently,the independent risk factors of AKI in patients after intra-coronary stent implantation could be found.Further,the prognosis of AKI patients was analyzed.Results Of the 325 patients,51 (15.7％) developed AKI.Compared the normal group,hospital stay (P ＜ 0.01 ) and in-hospital mortality (P ＜ 0.05) increased significantly in the AKI group.Monofactorial analysis showed that age,pre-operative laboratory and clinical data including left ventricular insufficiency,peripheral angiopathy,creatinine,urea nitrogen,estimated glomerular filtration rate, hyperuricemia, proteinuria, hydration and emergency operation, and intraoperative information such as operation time and hypotension,and postoperative hypotension in AKI patients group were significantly different in comparison with control group ( P ＜ 0.05 ). Multivariate logistic regression analysis revealed that elderly age (OR =0.253),pre-operative proteinuria (OR =5.351 ),preoperative left ventricular insufficiency ( OR =8.704),eGFR ≤ 60 ml/ ( min · 1.73 m2 ) ( OR =6.677 ),prolonged operation time ( OR =1.017),intra-operative hypotension ( OR =25.245 ) were independent risk factors of AKI ( P ＜ 0.05 ).Conclusions AKI is a common complication and associated with increase in mortality after intra-coronary stent implantation.Increase in age,pre-operative proteinuria,pre-operative left ventricular insufficiency,pre-operative low estimated glomerular filtration rate,prolonged operation time,intra-operative hypotension are the independently risk factors associated with AKI.

Objective To investigate the features of risk factors of minor stroke with CISS classification in Guangdong Province. Methods We retrospectively investigated the patients admitted within 3 days of the occurrence of a minor stroke, and were classified by CISS criteria as large artery atherosclerosis (LAA), cardiogenic stroke (CS), penetrating artery disease (PAD), other etiology (OE), undetermined etiology (UE). Results In this study, 303 pa-tients met the inclusion criteria of minor stroke. The highest percentage of the risk factors included hypertension (72.3%), hyperlipidemia (58.3%), and diabetes mellitus (39.9%). Among different subtypes, 41.9% were diagnosed with LAA, and 50.8% with PAD. Plasma triglyceride (TG)(1.765 ±1.18)mg/L vs.(2.19 ±1.84)mg/L,P=0.03], apolipoproteinsB (ApoB) [(0.95±0.29)mg/L vs.(1.11±0.46)mg/L,P=0.009]C-reactive protein (CRP) [(6.63±11.30) mg/L vs.(3.42 ±5.02)mg/L,P=0.042] and ApoB/ApoA1 ratio [(0.754 ±0.25)mg/L vs.(0.875 ±0.49)mg/L,P=0.019], differed significantly between group LAA and PAD. Conclusion Hypertension, hyperlipidemia and diabetes mellitus are the major risk factors of minor stroke. The most common subtypes of the minor stroke patients in Guangdong Province are LAA and PAD, and detecting their TG, apoB, CRP level and apoB/apoA1 ratio might help subclassify minor stroke according to CISS.

ObjectiveGeneration of human induced pluripotent stem cells from human skin fibroblasts.MethodsSox2, Klf4, Oct4, c-Myc were transfected into HSF cells with retrovirus, and then HSF cells was reprogrammed to iPS cells.Detecting cells endogenous and exogenous gene, analyzing karyotype,cells alkaline phosphatase staining and immunofluorescence staining, differentiating into teratomas in vivo and embryoid bodies in vitro were performed.ResultsiPS cell morphology was similar to embryonic stem cells (ES).The expression of Nanog, Oct4, Rex1, Sox2 in iPS cells were higher than HSF cells, and Sox2, Klf4, Oct4, c-Myc were silenced for the iPS cells.Exogenous genes were inserted into the nucleus of iPS cells, which was confirmed by 1％ agarose gel electrophoresis.iPS cell karyotype was normal, alkaline phosphatase activity increased, ES cell-specific protein expressed.iPS cells were differentiated into a teratoma in vivo and embryoid bodies in vitro.ConclusionsiPS cells were similar to ES cells, which have pluripotency.

Objective To evaluate the correlation between the degree of cochlea endolymphatic hydrops(EH) and hearing loss and symptoms in patients with unilateral Meniere's disease. Methods Fifty seven patients with unilateral Meniere's patients were retrospectively quantitatively analyzed, which evaluated the correlation between the cochlea EH and hearing loss and symptoms. The affected ears in the experimental group(57 ears) and the asymptomatic ears in the control group(57 ears), were confirmed by bilateral intratympanic Gd-DTPA injection and 3D real IR MRI scan after 24 h. The maximum length of endolymph space and labyrinth chamber along the modiolus cochleae and their ratio which represented the endolymph space proportion of each turn(R1, the basal turn;R2, the middle turn;R3, the apical turn) were calculated. And the paired t test was used to compare the differences in EH degree between the experimental and control group;Pearson correlation analysis was used to analyze the correlation between the cochlea EH degree and hearing loss and duration of symptoms. Results R1, R1 and R3 of ipsilateral cochlea were higher than normal cochlea(ipsilateral R1:0.354±0.097 vs. normal R1:0.185±0.031, P<0.01;ipsilateral R2:0.445 ± 0.098 vs. normal R2:0.201 ± 0.046, P<0.01;ipsilateral R3:0.467 ± 0.102 vs. normal R3:0.221 ± 0.053, P<0.01). A positive correlation was found between the degree of EH in each turn and high-frequency hearing loss(r=0.275, 0.281, 0.329, respectively;all P<0.05), whereas no correlation was found between the extent of EH and the time of vertigo, tinnitus and hearing loss(P>0.05). In addition, there was a correlation between the degree of EH in the apical turn and low, medium, high-frequency hearing loss(r=0.271, 0.269, 0.329, respectively; all P<0.05). Conclusions The degree of cochlea EH in the basal and second turn showed great relevance with the high-frequency hearing loss, and the apical turn EH degree was relevant with the low, medium, high-frequency hearing loss, but there was no correlation between the EH degree of each turn, ages and symptoms.