A data process of human knee joint kinematics obtained by motion-capture measurement.

BACKGROUND: The motion capture has been used as the usual method for measuring movement parameters of human, and most of the measuring data are obtained by partial manual process based on commercial software. An automatic kinematics data process was developed by programming on MATLAB software in this paper. METHODS: The motion capture measurement of healthy volunteers was carried out and the MATLAB program was used for data process. Firstly, the coordinate data of markers and anatomical points on human lower limb measured by motion capture system were read and repaired through the usual and the patch program. Meantime, the local coordinate systems of human femur and tibia were established with anatomical points. Then flexion/extension, abduction/adduction and internal/external rotation of human knee tibiofemoral joint were obtained by special coordinate transformation program. RESULTS: Using the above methods, motion capture measurements and batch data processing were carried out on squatting and climbing stairs of 29 healthy volunteers. And the motion characteristics (flexion/extension, internal/external rotation and adduction/abduction) of the knee joint were obtained. For example, the maximum internal/external rotation in squatting and climbing stairs were respectively was 30.5 degrees and 14 degrees, etc. Meantime, the results of this paper also were respectively compared with the results processed by other research methods, and the results were basically consistent, thus the reliability of our research method was verified. After calibration processing, the compiled MATLAB program of this paper can directly be used for efficient batch processing and avoiding manual modeling one by one. CONCLUSION: A novel Patch Program of this paper has been developed, which can make reasonable compensation for missing and noise signals to obtain more complete motion data. At the same time, a universal data processing program has also been developed for obtaining the relative movement of various components of the human body, and the program can be modified for detail special analysis. These motion capture technologies can be used to judge whether the human body functions are abnormal, provide a reference for rehabilitation treatment and design of rehabilitation equipment, and evaluate the effectiveness before and after surgery.

Isolation and characterization of polysaccharides with the antitumor activity from Tuber fruiting bodies and fermentation system.

Fifty-two polysaccharides were isolated from the fermentation systems of Tuber melanosporum, Tuber indicum, Tuber sinense, Tuber aestivum and the fruiting bodies of Tuber indicum, Tuber himalayense, Tuber sinense by elution with an activated carbon column. Polysaccharides from Tuber fermentation system exhibited relatively higher in vitro antitumor activity against HepG2, A549, HCT-116, SK-BR-3, and HL-60 cells than those from Tuber fruiting bodies. All polysaccharides were mainly composed of D-mannose, D-glucose, and D-galactose, which suggested that the polysaccharides from Tuber fruiting bodies and fermentation system have identical chemical compositions. The results of antitumor activity and structural identification indicated that the polysaccharide fractions could promote antitumor activity. Tuber polysaccharides from Tuber fermentation system exhibited relatively higher than that from Tuber fruiting bodies. These results confirm the potential of Tuber fermentation mycelia for use as an alternative resource for its fruiting bodies.

Artificial antigen synthesis and the development of polyclonal antibody-based immunoassay for citreoviridin determination.

Citreoviridin, a mycotoxin produced by Penicillium citreonigrum is a common contaminant of wide range of agri-products and detrimental to human and animal health. Therefore it is important to develop a rapid, sensitive, and specific immunoassay for citreoviridin detection. In this study, polyclonal antibody against citreoviridin was developed. For the preparation of citreoviridin-bovine serum albumin conjugate (CIT-BSA), hydroxyl groups on adjacent carbon atoms were oxidized by sodium periodate, so the product with reactive aldehyde residues was suitable for coupling with amine. Anti-citreoviridin polyclonal antibody was prepared by immunizing mice with CIT-BSA conjugate. The specificity and sensitivity of the polyclonal antibody was determined by indirect competitive ELISA. Results showed that the IC50 value of the polyclonal antibody was 0.56 µg/mL and no cross-reactivity was found between antiserum and other mycotoxins used in the experiment. The citreoviridin recovery rates by this polyclonal antibody were calculated through rice powder spiked by artificial citreoviridin. The recovery rates ranged were found from 70.5 ± 0.08 % to 94.7 ± 0.09% for inter-assay, and from 77.5 ± 0.04% to 95.4 ± 0.18% for intra-assay, which indicated that this polyclonal antibody could detect trace amount of CIT from the tested samples. Consequently, this study provided a specific and sensitive anti-citreoviridin polyclonal antibody, which made the determination of citreoviridin easier, quicker, and more accurate.

Potent in vitro synergism of fusidic acid (FA) and berberine chloride (BBR) against clinical isolates of methicillin-resistant Staphylococcus aureus (MRSA).

It was found in the present study that combined use of fusidic acid (FA) and berberine chloride (BBR) offered an in vitro synergistic action against 7 of the 30 clinical methicillin-resistant Staphylococcus aureus (MRSA) strains, with a fractional inhibitory concentration (FIC) index ranging from 0.5 to 0.19. This synergistic effect was most pronounced on MRSA 4806, an FA-resistant isolate, with a minimum inhibitory concentration (MIC) value of 1,024 µg/ml. The time-kill curve experiment showed that FA plus BBR yielded a 4.2 log10 c.f.u./ml reduction in the number of MRSA 4806 bacteria after 24-h incubation as compared with BBR alone. Viable count analysis showed that FA plus BBR produced a 3.0 log10 c.f.u./ml decrease in biofilm formation and a 1.5 log10 c.f.u./ml decrease in mature biofilm in viable cell density as compared with BBR alone. In addition, phase contrast micrographs confirmed that biofilm formation was significantly inhibited and mature biofilm was obviously destructed when FA was used in combination with BBR. These results provide evidence that combined use of FA and BBR may prove to be a promising clinical therapeutic strategy against MRSA.

YAP promotes the early development of temporomandibular joint bony ankylosis by regulating mesenchymal stem cell function.

To explore the role of YAP, a key effector of the Hippo pathway, in temporomandibular joint (TMJ) ankylosis. The temporal and spatial expression of YAP was detected via immunohistochemistry and multiplex immunohistochemistry on postoperative Days 1, 4, 7, 9, 11, 14 and 28 in a sheep model. Isolated mesenchymal stem cells (MSCs) from samples of the Day 14. The relative mRNA expression of YAP was examined before and after the osteogenic induction of MSCs. A YAP-silenced MSC model was constructed, and the effect of YAP knockdown on MSC function was examined. YAP is expressed in the nucleus of the key sites that determine the ankylosis formation, indicating that YAP is activated in a physiological state. The expression of YAP increased gradually over time. Moreover, the number of cells coexpressing of RUNX2 and YAP-with the osteogenic active zone labelled by RUNX2-tended to increase after Day 9. After the osteogenic induction of MSCs, the expression of YAP increased. After silencing YAP, the osteogenic, proliferative and migratory abilities of the MSCs were inhibited. YAP is involved in the early development of TMJ bony ankylosis. Inhibition of YAP using shRNA might be a promising way to prevent or treat TMJ ankylosis.

Oncologic safety of the pedicled submental island flap for reconstruction in oral tongue squamous cell carcinoma: An analysis of 101 cases.

OBJECTIVE: To evaluate whether the pedicle submental island flap (SIF) can be safely used in the oral tongue squamous cell carcinoma (OTSCC) patients with pathologically node-positive (pN+) neck, especially pN+ at level I. METHODS: Retrospectively, 101 OTSCC patients with SIF reconstruction were enrolled. Oncological outcomes included the total locoregional recurrence, the SIF related locoregional recurrence (SRLR) which referred to the local recurrence at flap and ipsilateral neck recurrence at level I, recurrence free survival (RFS), overall survival (OS), and disease specific survival (DSS). RESULTS: Sixty-one patients were pathologically node-negative (pN0) and 40 were pN+. Thirteen patients experienced locoregional recurrence, of which 5 had a SRLR. The pN+ group had a significantly higher locoregional recurrence rate, lower 5-year RFS, OS and DSS than pN0 group (P < 0.05). Patients with pN0 had a significantly higher neck RFS when compared to those with pN+ either at level I (P = 0.005) or at other levels (P < 0.001). However, the neck RFS was similar between the two subgroups of pN+ (P = 0.550). Especially, patients with pN+ at level I had a significantly higher SRLR rate (P = 0.006) compared to those with pN0 at level I. Multivariate analysis showed that pN+ was an unfavorable factor for tumor recurrence and OS. CONCLUSION: Our data did not support the use of SIF in OTSCC patients with pN+ neck at level I due to an significantly increased SRLR rate compared to those with pN0 neck at level I.

CWI pathway participated in vegetative growth and pathogenicity through a downstream effector AflRlm1 in Aspergillus flavus.

The cell wall is an essential dynamic structure for shielding fungus from environmental stress, and its synthesizing and remodeling are regulated by the cell wall integrity (CWI) pathway. Here, we explored the roles of a putative downstream effector AflRlm1 of CWI pathway in Aspergillus flavus. The results showed that AflRlm1 played a positive role in conidia production, sclerotium formation, aflatoxin biosynthesis, and pathogenicity. Furthermore, we provided evidence for the physical connection between AflRlm1 and AflSlt2 and determined the role of AflSlt2 in the phosphorylation of AflRlm1. Then, we discovered the importance of WSCs (cell wall integrity and stress response component) to the CWI signal and the process of AflRlm1 transferring to the nucleus after receiving the signal. Overall, this study clarified the transmission process of CWI signals and proves that the CWI pathway plays a key role in the development of A. flavus and the production of aflatoxin combined with transcriptome data analysis.

Functional analysis of an alpha-1,2-mannosidase from Magnaporthe oryzae.

Identification of enzymes that are expressed during host colonization and characterization of their biochemical properties are prerequisite to understanding their role in the pathogen-host interaction. Nine alpha-1,2-mannosidase homologs were identified in the analysis of the Magnaporthe oryzae genome. Endoplasmic reticulum localized alpha-1,2-mannosidases play an important role in protein glycosylation. However, several members of the alpha-1,2-mannosidase gene family are predicted to be secreted. The biological role of such extracellular enzymes in host colonization has not been defined. Here, we characterized a secreted alpha-1,2-mannosidase of M. oryzae, MGG_00994.6, and found that the mature polypeptide is a glycoprotein capable of hydrolyzing alpha-1,2 linked mannobiose. The gene is expressed during growth in vitro and during colonization on rice plants, however, deletion of the gene did not affect pathogenicity. Five other members of the alpha-1,2-mannosidase of M. oryzae were expressed with a pattern similar to MGG_00994.6, suggesting the potential for functional redundancy. These results form the basis for additional studies on the role of this gene family in the rice blast fungus and its interaction with rice.

2-Amino-nonyl-6-methoxyl-tetralin muriate inhibits sterol C-14 reductase in the ergosterol biosynthetic pathway.

AIM: To investigate the action mechanism of a novel chemical structural aminotetralin derivate, 2-Amino-Nonyl-6-Methoxyl-Tetralin Muriate (10b), against Candida albicans (C albicans) in the ergosterol biosynthetic pathway. METHODS: Antifungal susceptibility test of 10b was carried out using broth microdilution method, the action mechanism of 10b against C albicans was investigated by GC-MS spectrometry and real-time RT-PCR assay, and cytotoxicity of 10b in vitro was assessed by MTS/PMS reduction assay. RESULTS: 10b reduced the ergosterol content markedly, and the 50% ergosterol content inhibitory concentration (ECIC(50) value) was 0.08 microg/mL. Although the sterol composition of 10b-grown cells was completely identical with that of erg24 strain, the content of ergosta-8,14,22-trienol in 10b-grown cells was much higher than that in erg24 strain. Real-time RT-PCR assay revealed a global upregulation of sterol metabolism genes. In addition, the 50% inhibitory concentration (IC(50) value) of 10b was 11.30 microg/mL for murine embryonic fibroblasts and 35.70 microg/mL for human normal liver cells. CONCLUSION: 10b possessed a mode of action different from that of azoles and morpholines, whose targets were sterol C-14 reductase (encoded by ERG24 gene) and sterol C-5 desaturase (encoded by ERG3) related enzyme. Although 10b seemed to reduce MTS/PMS reduction in a dose dependent manner, IC(50) value for mammalian cells was much higher than 50% minimum inhibitory concentration (MIC(50)) value for C albicans. This indicates that the formulation is preliminarily safe and warrants further study for possible human applications.

Structural stability of a polyetheretherketone femoral component-A 3D finite element simulation.

BACKGROUND: Because its mechanical properties are similar to cortical bones of the knee, polyetheretherketone (PEEK) material has been used to make total knee arthroplasty (TKA) components. This study investigated the PEEK femoral component deformation of a TKA system and compared the data with that of a cobalt-chromium (CoCr) component. METHODS: A 3D finite element knee model was constructed using CT images of a normal subject. A knee prosthesis was installed on the model to simulate a TKA knee. The material properties of the bone were assumed linear and transverse isotropic. The femoral component was modeled using a PEEK or CoCr material. A compressive load was applied to the knee at full extension. Tibiofemoral contact stresses and femoral component deformations were analyzed. FINDINGS: Under a 3 kN load, the maximal Von-Mises stresses in the femoral component were 14.39 MPa and 30.05 MPa for the PEEK and CoCr components, respectively. At the tibial polyethylene surface, the CoCr femoral component caused higher contact stresses (>2.2%) than the PEEK component. The deformation of the PEEK component was over 3 times larger than that of the CoCr component (0.65 × 10-3 mm vs 0.2 × 10-3 mm). INTERPRETATION: The PEEK femoral component could result in lower contact stresses, but larger deformations in the TKA knee compared to the CoCr component. An increased deformation of the PEEK component indicates a reduction in its structural strength. Future investigation should examine if the reduced structural strength will affect the in-vivo component-bone interface integration and affect the component fatigue life.

[Expression and function of microRNA in embryonic stem cell].

Embryonic stem cells are the cells that have the ability both to self-renew and to differentiate into all the mature cell types in an adult. Both of these processes are tightly regulated by genetic and epigenetic mechanisms. Increasing evidence indicates that a class of single-stranded non-coding RNA, known as "microRNAs", also plays a critical role in this process. MicroRNA can bind to target mRNAs by specific base pairing, then degrade mRNAs or inhibit protein translation. Therefore, they can participate in post-transcriptional regulation. Recently, scientists began to study the effect of microRNA on embryonic stem cell and found that some microRNAs are specifically expressed and form an intricate network of microRNAs, regulating key transcription factors and other genes. This review focuses on the expression of microRNA in embryonic stem cell and their functions. We discuss some microRNA that are specifically expressed in embryonic stem cell and their regulating effect on self-renewal and differentiation.

Detection of deoxynivalenol based on a single-chain fragment variable of the antideoxynivalenol antibody.

Immunological detection of low molecular weight toxins, such as deoxynivalenol using single-chain antibody fragment (scFv), is a potentially novel and safe method of diagnosing fungal infection and food contamination. To develop a deoxynivalenol detection procedure based on scFv, deoxynivalenol was first conjugated to horseradish peroxidase (HRP) for immunoassay. Deoxynivalenol was initially activated using N-[p-maleimidophenyl] isocyanate and subsequently conjugated to S-acetyl thioglycolic acid N-hydroxysuccinimide-activated HRP. Sodium dodecylsulphate polyacrylamide gel electrophoresis and enzyme-linked immunosorbent assay (ELISA) analysis confirmed covalent attachment of deoxynivalenol to HRP successfully. Competitive direct-ELISA based on antideoxynivalenol scFv was used to detect deoxynivalenol in spiked and natural contaminated wheat samples, and the results showed that scFv is applicable in deoxynivalenol detection of contaminated wheat samples. This work confirms that sensitive and specific scFv against fungal toxins can be applicable in diagnosis of fungal infection.

Effects of Qindan Capsule on blood pressure, endothelin, calcitonin gene-related peptide and angiotensin- II in spontaneous hypertensive rats.

OBJECTIVE: To observe the hypotensive effects of Qindan Capsule (QC) on spontaneous hypertensive rats (SHR) and its effect on the contents of endothelin (ET), calcitonin gene-related peptide (CGRP) and angiotensin-II (Ang-II ) in plasma and vascular tissues, and to investigate the possible mechanism of QC in lowering blood pressure. METHODS: Forty SHRs were divided into 5 groups: the high dosage QC group [QCHD, 750 mg/(kgxd)], the low dosage QC group [QCLD, 150 mg/(kgxd) ], the Niuhuang Jiangya Pill group [NJP, 200 mg/(kgxd) ], the Captopril group [ 15 mg/(kg d) land the model group, 8 in each group. Meanwhile, a normal control group consisting of 8 Wistar-Kyoto (WKY) rats was set up also. All the rats were administered with medicine through gastrogavage. Systolic blood pressure (SBP), level of ET, CGRP and Ang-II in plasma and Ang-II in tissues of mesenteric artery were detected in all the rats after 12 weeks of treatment. RESULTS: The level of SBP after treatment in the QCHD group was lower than that in the model group (P<0.01), but with no significant difference as compared with that in the Captopril group and the NJP group (P>0.05). After treatment, the plasma level of ET was lower and CGRP higher than those in the model group (both P<0.05), and also higher than those in the NJP and Captopril group (both P<0.05). As for the content of Ang-II , in mesenteric arterial tissues, it was lower in the QCHD group than that in the model group ( P<0.05), but in plasma, it showed no significant difference between the two groups (P>0.05). CONCLUSION: QC has a satisfactory hypotensive action on SHR rats, and its mechanism may be associated with the regulation on plasma vasoactive peptide and regional renin-angiotensin system.

PMD: A Resource for Archiving and Analyzing Protein Microarray data.

Protein microarray is a powerful technology for both basic research and clinical study. However, because there is no database specifically tailored for protein microarray, the majority of the valuable original protein microarray data is still not publically accessible. To address this issue, we constructed Protein Microarray Database (PMD), which is specifically designed for archiving and analyzing protein microarray data. In PMD, users can easily browse and search the entire database by experimental name, protein microarray type, and sample information. Additionally, PMD integrates several data analysis tools and provides an automated data analysis pipeline for users. With just one click, users can obtain a comprehensive analysis report for their protein microarray data. The report includes preliminary data analysis, such as data normalization, candidate identification, and an in-depth bioinformatics analysis of the candidates, which include functional annotation, pathway analysis, and protein-protein interaction network analysis. PMD is now freely available at www.proteinmicroarray.cn.

Identification and characterization of Bacillus anthracis by multiplex PCR on DNA chip.

Bacillus anthracis can be identified by detecting virulence factor genes located on two plasmids, pXO1 and pXO2. Combining multiplex PCR with arrayed anchored primer PCR and biotin-avidin alkaline phosphatase indicator system, we developed a qualitative DNA chip method for characterization of B. anthracis, and simultaneous confirmation of the species identity independent of plasmid contents. The assay amplifies pag gene (in pXO1), cap gene (in pXO2) and Ba813 gene (a B. anthracis specific chromosomal marker), and the results were indicated by an easy-to-read profile based on the color reaction of alkaline phosphatase. About 1 pg of specific DNA fragments on the chip wells could be detected after PCR. With the proposed method, the avirulent (pXO1+/2-, pXO1-/2+ and pXO1-/2-) strains of B. anthracis and distinguished 'anthrax-like' strains from other B. cereus group bacteria were unambiguously identified, while the genera other than Bacillus gave no positive signal.

Pharmacological blockade of the MaxiK channel attenuates experimental acute pancreatitis and associated lung injury in rats.

Increasing evidence has recently demonstrated that soluble heparan sulfate (HS), a degradation product of extracellular matrix produced by elastase, plays a key role in the aggravation of acute pancreatitis (AP) and associated lung injury. However little is known about the detailed mechanism underlying HS-induced inflammatory cascade. Our previous work has provided a valuable clue that a large-conductance K(+) channel (MaxiK) was involved in the HS-stimulated activation of murine macrophages. Here we attempted to ask whether pharmacological inhibition of the MaxiK channel will exert beneficial effects on the treatment of AP and secondary lung injury. The protective effects of paxilline, a specific blocker of MaxiK, on rats against sodium taurocholate induced AP were evaluated. Our data showed that paxilline substantially attenuated AP and resultant lung injury, mainly by limiting the burst of inflammatory responses, as proven by decreased plasma concentrations of tumor necrosis factor-α and macrophage inflammatory protein-2, together with unimpaired pancreatic enzyme activities in rats suffering from AP. Compared with the therapeutic administration, pre-treatment of paxilline showed superior potential to slow down the progress of AP. Furthermore, AP rats received paxilline exhibited improved histopathologic alterations both in the pancreas and the lungs, and even lower lung MPO activity. Taken together, our study provides evidence that MaxiK is involved in the spread of inflammatory responses and the following lung injury during the attack of AP, indicating that this ion channel is a promising candidate as a therapeutic target for AP.

Allogeneic bone marrow mesenchymal stem cell transplantation in patients with UDCA-resistant primary biliary cirrhosis.

The objective of this study was to evaluate the safety and efficacy of allogeneic bone marrow mesenchymal stromal/stem cell transplantation (BM-MSCT) for patients with ursodeoxycholic acid (UDCA)-resistant primary biliary cirrhosis (PBC). Ten patients were enrolled in this trial of BM-MSCT. All patients were permitted to concurrently continue their previous UDCA treatment. The efficacy of BM-MSCT in UDCA-resistant PBC was assessed at various time points throughout the 12-month follow up. No transplantation-related side effects were observed. The life quality of the patients was improved after BM-MSCT as demonstrated by responses to the PBC-40 questionnaire. Serum levels of ALT, AST, γ-GT, and IgM significantly decreased from baseline after BM-MSCT. In addition, the percentage of CD8+ T cells was reduced, while that of CD4+CD25+Foxp3+ T cells was increased in peripheral lymphocytic subsets. Serum levels of IL-10 were also elevated. Notably, the optimal therapeutic outcome was acquired in 3 to 6 months and could be maintained for 12 months after BM-MSCT. In conclusion, allogeneic BM-MSCT in UDCA-resistant PBC is safe and appears to be effective.

2-amino-nonyl-6-methoxyl-tetralin muriate activity against Candida albicans augments endogenous reactive oxygen species production --a microarray analysis study.

Candida infections have become an increasingly significant problem, mainly because of the widespread nature of Candida and drug resistance. There is an urgent need to develop new classes of drugs for the treatment of opportunistic Candida infections, especially in medically complex patients. Previous studies have confirmed that 2-amino-nonyl-6-methoxyl-tetralin muriate (10b) possesses powerful antifungal activity in vitro against Candia albicans. To clarify the underlying action mechanism, an oligonucleotide microarray study was performed in C. albicans SC5314 without and with 10b treatment. The analytical results showed that energy metabolism-related genes, including glycolysis-related genes (PFK1, CDC19 and HXK2), fermentation-related genes (PDC11, ALD5 and ADH1) and respiratory electron transport chain-related genes (CBP3, COR1 and QCR8), were downregulated significantly. Functional analysis revealed that 10b treatment increased the generation of endogenous reactive oxygen species, and decreased mitochondrial membrane potential, ubiquinone-cytochrome c reductase (complex III) activity and intracellular ATP levels in C. albicans SC5314. Also, addition of the antioxidant ascorbic acid reduced the antifungal activity of 10b significantly. These results suggest that mitochondrial aerobic respiration shift and endogenous reactive oxygen species augmentation might contribute to the antifungal activity of 10b against C. albicans. This information may prove to be useful for the development of new strategies to treat Candida infections.

Construction of single chain variable fragment (ScFv) and BiscFv-alkaline phosphatase fusion protein for detection of Bacillus anthracis.

This paper describes an attempt for convenient and sensitive detection of Bacillus anthracis with single chain variable fragment (scFv)-based protein chip. Phage display technology was employed to generate scFv by using the protective antigen (PA) of B. anthracis for immunization. V(H) and V(L) genes of the scFv were amplified separately by reverse transcriptase-PCR from mRNA of immunized mice and then assembled into scFv gene with a linker DNA sequence. The scFv gene was inserted into a phagemid vector pCANTAB-5E and then transformed into Escherichia coli TG1 to yield recombinant phages after infection with helper phage M13KO7. After six rounds of panning with PA, the phage clones displaying scFv fragments of the antibody were selected by ELISA. One phage clone scFv-6w10 showing the strongest positive signal in ELISA was selected. To enhance the affinity of the scFv-6w10, a recombinant bivalent single-chain Fv antibody (biscFv-6w10) directed against PA was constructed and tested in functional assays. The affinity of the biscFv-6w10 was much higher than that of scFv-6w10 and reached 6.5 x 10(9) M(-1). An expression system was constructed for the production of E. coli alkaline phosphatase (EAP) labeled biscFv-6w10 (biscFv-6w10-EAP) in E. coli cells. The expressed fusion protein retained both antigen-specific binding and enzymatic activity and thus directly served as an enzyme-labeled antibody. Detections of PA and bacterial cells of B. anthracis using biscFv-6w10-EAP and Cy3-labeled biscFv-6w10 were performed on a protein chip. The fusion protein (biscFv-6w10-EAP) chip could detect 10 pg of PA and 500-1000 bacterial cells in approximately 2 h, while the sensitivity of Cy3-labeled protein chip reached 1 pg of PA and 50-100 cells within 2 h.