Research on the Method of Acquiring Customer Individual Demand Based on the Quantitative Kano Model.

In order to realize accurate marketing by analyzing customer individual demand, a new quantitative Kano model method is put forward, and it is helpful to provide customized products for heterogeneous customer classification groups. By improving the traditional Kano model, the customer satisfaction and the importance degree of products are defined, and the quantitative Kano demand model is established. Customers are classified as the price preference group, the brand preference group, and the service priority group, and decision-making of product attribute quality improvement for customer classification is realized. Lastly, electric vehicles (EVs) are selected as a study case, and their various demands for different classifications of customers are discussed by questionnaire survey and calculation of satisfaction and the importance degree. Furthermore, different customer group demands are classified as attractive demands, expected demands, nondifferential demands, or essential demands, and the important product attribute acquisition process for various customers is discussed to improve enterprise market competitiveness.

Medical image management and analysis system based on web for fungal keratitis images.

The medical image management and analysis system proposed in this paper is a medical software developed by the Browser/Server (B/S) architecture after investigating the workflow of the relevant departments of the hospital, which realizes the entire process of patients from consultation to printing of reports. The computer-aided diagnosis function is added based on image management. Due to the difficulty in collecting medical image data, in the computer-aided diagnosis module, this paper only uses the common fungal keratitis collected from the hospital in the laboratory. Focused microscope images are used for experiments. First, the images were trained with three convolutional neural networks, AlexNet, ZFNet, and VGG16. These models which classify fungal keratitis were obtained and integrated was performed to obtain better classification results. Finally, the model was integrated with the system designed in this paper, which realized the automatic diagnosis of Confocal Microscopy (CM) images of fungal keratitis online and provided it to medical staff for reference. The system can improve the work efficiency of the image-related departments while reducing the workload of doctors in the department to manually read the films.

Detection of carbapenemase-encoding genes among clinical isolates of Pseudomonas aeruginosa in a Chinese burn unit.

The carbapenemases have recently emerged as molecules implicated in one of the most feared bacterial resistance mechanisms because of their ability to hydrolyze virtually all lactamase agents and their highly mobile genes. This study aimed to investigate the prevalence of carbapenemase and antimicrobial susceptibilities of Pseudomonas aeruginosa isolated from burn patients in Chongqing, China. Antimicrobial susceptibility of 111 isolates was determined by the disc agar diffusion test and the agar dilution method. Random Amplification of Polymorphic DNA polymerase chain reaction analysis revealed 111 P. aeruginosa 42 genotypes. Carbapenemase genes were amplified by polymerase chain reaction and the sequence verified by blast. Ninety-three of 111 (83.8%) isolates were resistant to imipenem; all of them had developed multidrug resistance and exhibited higher resistant rates compared with the imipenem-susceptible Pseudomonas. Ciprofloxacin was the most effective antipseudomonal agent. Thirty-three of the isolates were identified to contain the metallo-ß-lactamase blaIMP-4 gene and belong to different Random Amplification of Polymorphic DNA polymerase chain reactiongenotypes. In conclusion, the high prevalence of multidrug resistance (94.6%) and the production of blaIMP-4 genes in P. aeruginosa isolates in burn patients highlight the necessity of considering these issues in burn hospitals.

Emulsified isoflurane postconditioning produces cardioprotection against myocardial ischemia-reperfusion injury in rats.

Emulsified isoflurane (EIso) preconditioning can induce cardioprotection. We investigated whether EIso application after ischemia protects hearts against reperfusion injury and whether it is mediated by the inhibition of apoptosis. Rats were subjected to 30-min coronary occlusion followed by 180-min reperfusion. At the onset of reperfusion, rats were intravenously administered saline (sham, control group), 30 % intralipid (IL group) or 2 ml kg(-1) EIso (EIso group) for 30 min. After reperfusion, infarct sizes, myocardial apoptosis and expression of Bcl-2, Bax and caspase-3 proteins were determined. Hemodynamic parameters were not different among groups. Compared with control and intralipid group, EIso limited infarct size, inhibited apoptosis, increased the expression of Bcl-2, decreased the expression of Bax, cleaved caspase-3, and enhanced Bcl-2/Bax ratio. EIso protects hearts against reperfusion injury when administered at the onset of reperfusion, which may be mediated by the inhibition of apoptosis via modulation of the expression of pro- and anti-apoptotic proteins.

Identification of mitochondria translation elongation factor Tu as a contributor to oxidative damage of postburn myocardium.

Mitochondrial damage plays an important role in mediating postburn cardiac injury. To elucidate the pivotal mitochondrial proteins and pathways underlying postburn cardiac injury, mitochondria were purified from control and postburn rat hearts. 2-dimensional gel electrophoresis (2-DE) and HPLC-chip-MS/MS analyses revealed 9 differentially expressed proteins, 3 of which were further validated by Western blotting. The differential expression of these mitochondrial proteins was accompanied by increased levels of oxidative cardiac damage and decreased levels of cardiac output. One of the differentially expressed proteins, mitochondria translation elongation factor Tu (EF-Tumt), was hypothesized to contribute crucially to postburn oxidative cardiac damage. The small interfering RNA (siRNA)-mediated downregulation of EF-Tumt in cultured rat cardiomyocytes increased reactive oxygen species (ROS) generation and protein carbonyl levels, and led to cell damage. The potential pathway of this process was associated with respiratory chain complex I deficiency. Together, these results demonstrate the mitochondrial responses to severe burn, and indicate a pathway by which decreased EF-Tumt expression mediates oxidative damage in postburn myocardium.

Spectrum and drug resistance of pathogens from patients with burns.

Microbial infection is an obstacle of burn treatment. However, little is known on what types of microbial infection dominate in the burn center and how the dynamic change of those microorganisms occurs during the past several years in China. We conducted a retrospective study of nosocomial infection (NI) in a large burn center to analyze the spectrum and antimicrobial resistance of microbial isolates from January 2003 to December 2010. We studied 989 isolates from 677 patients who had signs and symptoms of infection 48h after admission. The number of NIs per 100 admissions was 10.9. The commonest isolates were Pseudomonas aeruginosa (23.1%), Staphylococcus aureus (18.7%), and Candida (11.4%). The result indicated that the numbers of patients with Acinetobacter sp. infection increased (P=0.004), but with Proteus mirabilis infection decreased (P=0.004). The isolated Acinetobacter sp. and P. aeruginosa were consistently highly resistant to almost all antibiotics tested. Notably, more frequent Acinetobacter sp. isolates appeared to be resistant to amikacin, gentamicin, tobramycin, ceftazidim, piperacillin, tazobactam, levofloxacin, and ciprofloxacin and more frequent Escherichia coli isolates were resistant to ceftazidime and aztreonam at the late time period although the P. aeruginosa and E. coli isolates were sensitive to less used ciprofloxacin and piperacillin/tazobactam. The increased rates of drug-resistant isolates in the later period might be associated with regular prophylactic therapy with antibiotics.

MAP4 mechanism that stabilizes mitochondrial permeability transition in hypoxia: microtubule enhancement and DYNLT1 interaction with VDAC1.

Mitochondrial membrane permeability has received considerable attention recently because of its key role in apoptosis and necrosis induced by physiological events such as hypoxia. The manner in which mitochondria interact with other molecules to regulate mitochondrial permeability and cell destiny remains elusive. Previously we verified that hypoxia-induced phosphorylation of microtubule-associated protein 4 (MAP4) could lead to microtubules (MTs) disruption. In this study, we established the hypoxic (1% O(2)) cell models of rat cardiomyocytes, H9c2 and HeLa cells to further test MAP4 function. We demonstrated that increase in the pool of MAP4 could promote the stabilization of MT networks by increasing the synthesis and polymerization of tubulin in hypoxia. Results showed MAP4 overexpression could enhance cell viability and ATP content under hypoxic conditions. Subsequently we employed a yeast two-hybrid system to tag a protein interacting with mitochondria, dynein light chain Tctex-type 1 (DYNLT1), by hVDAC1 bait. We confirmed that DYNLT1 had protein-protein interactions with voltage-dependent anion channel 1 (VDAC1) using co-immunoprecipitation; and immunofluorescence technique showed that DYNLT1 was closely associated with MTs and VDAC1. Furthermore, DYNLT1 interactions with MAP4 were explored using a knockdown technique. We thus propose two possible mechanisms triggered by MAP4: (1) stabilization of MT networks, (2) DYNLT1 modulation, which is connected with VDAC1, and inhibition of hypoxia-induced mitochondrial permeabilization.

Role of serum amyloid P in skin graft survival and wound healing in burned patients receiving skin grafts.

BACKGROUND: Recent studies in animal models suggest that serum amyloid P (SAP) can affect burn wound healing. However, the role of SAP in a clinical setting remains unknown. METHODS: We enrolled 88 patients with third degree burn wounds. All the patients were candidates for auto-skin graft procedure using stamp skin graft. The complete graft healing time and the number of survived grafts were recorded. Serum SAP levels were assessed 1 day before operation. RESULTS: There was no significant difference in SAP level between controls and patients. There were no significant differences noted among the patients with different burn surface area. However, when the patients in each group were stratified by SAP levels, the mean complete healing time of grafted wound and the mean numbers of survived skin grafts were significantly different. Spearman's analyses showed that the serum SAP levels negatively correlated with the complete wound healing time and mean numbers of survived skin grafts. Logistic regression analysis showed that the serum SAP levels and mean numbers of survived skin grafts were potent independent factors contributing to wound healing. CONCLUSIONS: The results of this study suggest that the serum SAP levels may be an easy detected predictor for the healing of burn wounds.

[Influence of microtubule depolymerization of myocardial cells on mitochondria distribution and energy metabolism in adult rats].

OBJECTIVE: To investigate the influence of microtubule depolymerization of myocardial cells on distribution and activity of mitochondria, and energy metabolism of cells in adult rats. METHODS: Myocardial cells of SD adult rats and SD suckling rats were isolated and cultured. They were divided into adult and suckling rats control groups (AC and SC, normally cultured without any stimulating factor), adult and suckling rats microtubule depolymerization agent groups (AMDA and SMDA, cultured with 8 micromol/L colchicine containing nutrient solution for 30 minutes) according to the random number table. (1) The expression of polymerized beta tubulin in myocardial cells of adult and suckling rats was detected with Western blot. (2) Myocardial cells of rats in AC and AMDA groups were collected. The expression of cytochrome c was detected with Western blot. Distribution of voltage-dependent anion channels (VDAC) and polymerized beta tubulin in myocardial cells were observed with immunofluorescent staining. Mitochondrial inner membrane potential was determined with immunocytochemical method. Activity of myocardial cells was detected with MTT method. Contents of ATP, adenosine diphosphate (ADP), and adenosine monophosphate (AMP) and energy charge of cells were determined with high performance liquid chromatography. RESULTS: (1) The expression of polymerized beta tubulin:in AMDA group it was 0.52 + or - 0.07, which was obviously lower than that (1.25 + or - 0.12) in AC group (F = 31.002, P = 0.000); in SMDA group it was 0.76 + or - 0.12, which was significantly lower than that (1.11 + or - 0.24) in SC group (F = 31.002, P = 0.000), but was obviously higher than that in AMDA group (F = 31.002, P = 0.009). (2) The expression of cytochrome c in AC group was 0.26 + or - 0.03, which was obviously lower than that (1.55 + or - 0.13) in AMDA group (t = -24.056, P = 0.000). (3) Immunofluorescent staining result: in AC group, microtubules of myocardial cells were in linear tubiform, distributed in parallel with myocardial fiber; VDAC staining result showed that mitochondria were in granular form, distributed in the same direction as microtubules. In AMDA group, the normal distribution regularity of microtubules was destroyed, with weakened immune fluorescence intensity, microtubules structure indistinct, continuity lost, rough in appearance, and the distribution of mitochondria became disrupted. (4) Mitochondrial inner membrane potential in AC group fluorescent intensity was 1288 + or - 84, which was obviously higher than that (331 + or - 27) in AMDA group (t = 26.508, P = 0.000). (5) Cellular activity: in AC group absorbance value was 1.75 + or - 0.11, which was obviously lower than that (0.81 + or - 0.07) in AMDA group (t = 17.348, P = 0.000). (6) Energy metabolism: compared with those in AC group, content of ATP decreased, contents of ADP and AMP increased, and ATP/ADP value and energy charge decreased in AMDA group. CONCLUSIONS: Microtubules and mitochondria distribute in the same direction in normal myocardial cells in adult rats. After microtubule depolymerization, mitochondria are arranged in disorder fashion; cytochrome c leaks from mitochondria; mitochondrial membrane potential, energy supply, and cellular activity decrease in the myocardial cells.

Microtubular stability affects cardiomyocyte glycolysis by HIF-1alpha expression and endonuclear aggregation during early stages of hypoxia.

Hypoxia-inducible factor (HIF)-1alpha is a key regulator of anaerobic energy metabolism. We asked the following question: Does the breakdown of microtubular structures influence glycolysis in hypoxic cardiomyocytes by regulating HIF-1alpha? Neonatal rat cardiomyocytes were cultured under hypoxic conditions, while microtubule-stabilizing (paclitaxel) and -depolymerizing (colchicine) agents were used to change microtubular structure. Models of high microtubule-associated protein 4 (MAP4) expression and RNA interference of microtubulin expression were established. Microtubular structural changes and intracellular HIF-1alpha protein distribution were observed with laser confocal scanning microscopy. Content of key glycolytic enzymes, viability, and energy content of cardiomyocytes were determined by colorimetry and high-performance liquid chromatography. HIF-1alpha protein content and mRNA expression were determined by Western blotting and real-time PCR, respectively. Low doses of microtubule-stabilizing agent (10 mumol/l paclitaxel) and enhanced expression of MAP4 stabilized the reticular microtubular structures in hypoxic cardiomyocytes, increased the content of key glycolytic enzymes, ameliorated energy supply and enhanced cell viability, and upregulated HIF-1alpha protein expression and endonuclear aggregation. In contrast, the microtubule-depolymerizing agent (10 mumol/l colchicine) or reduced microtubulin expression had adverse affects on the same parameters, in particular, HIF-1alpha protein content and endonuclear aggregation. We conclude that microtubular structural changes influence glycolysis in the early stages of hypoxia in cardiomyocytes by regulating HIF-1alpha content. Stabilizing microtubular structures increases endonuclear and total HIF-1alpha expression, content of key glycolytic enzymes, and energy supply. These findings provide potential therapeutic targets for ameliorating cell energy metabolism during early myocardial hypoxia.

[The influence of microtubule intervention drugs on glycolytic key enzymes in myocardial cells after hypoxia].

OBJECTIVE: To investigate the influence of microtubule intervention drugs on glycolytic key enzymes in myocardial cells after hypoxia. METHODS: The primary passage of cultured myocardial cells from neonatal rats were divided into A group (with hypoxia), B group (with hypoxia and administration of l0 micromol/L colchicine), C group (with hypoxia and administration of 5 micromol/L taxol), D group (with hypoxia and administration of 10 micromol/L taxol), E group (with hypoxia and administration of 15 micromol/L taxol). The morphology of microtubule was observed with laser scanning microscope (LSM). The cell vitality was assayed by cell counting kit (CCK). The activities of hexokinase (HK), pyruvate kinase (PK), phosphofructokinase (PFK) and lactate dehydrogenase (LDH) were assayed with colorimetry. RESULTS: In group B and E, the microtubule structure was damaged heavily, and the cell vitality was decreased significantly [The cell vitality was (89.99 +/- 3.47)% in B group and (84.56 +/- 6.61)% in E group, respectively, at 1.0 post hypoxia hour (PHH), and hoth values were obviously lower than that in A group (97.44 +/- 1.76)%, P < 0.01]. The HK, PK and PFK activities decreased obviously. The activities of HK, PK and PFK in group C were similar to those of the A group. Compared with that in other groups, the degree of damage of microtubule structure in D group was milden. The activities of HK, PK and PFK in D group during 0.5 - 6.0 PHH were significantly higher than those in A group. The activity of LDH in each group was increased after hypoxia. CONCLUSION: Proper concentration of microtubule-stabilizing drugs can alleviate the damages to microtubule structure, and enhance the activity of glycolytic key enzymes of myocardial cells at early stage of hypoxia.

[Study on the influence of hypoxia induced microtubule damage on the opening of mitochondrial permeable transition pore of cardiac myocytes in rat].

OBJECTIVE: To investigate the influence of hypoxia induced microtubule damage on the opening of mitochondrial permeable transition pore (MPTP)of cardiac myocytes and on the decrease of respiratory function in rat. METHODS: Primary cultured myocardial cells from 30 neonatal rats were randomized as normoxic group (A), hypoxia group (B), normoxia with microtubule destabilizing agent group (C, with treatment of 8 micromol/L colchicines for 30 minutes before normoxia), and hypoxia with microtubule stabilizing agent group (D, with treatment of 10 micromol/L taxol for 30 minutes before hypoxia). beta-tubulin immunofluorescence ,the opening of mitochondria permeability transition pore, and the mitochondrial inner membrane potential were detected at 0.5, 1, 3, 6 and 12 post-treatment hours (PTH), and the mitochondrial respiratory function was determined by MTT method. The changes in these indices were also determined in A group at the corresponding time-points. RESULTS: Obvious damage of polymerized microtubule, opening of MPTP, mitochondrial inner membrane potential loss and decrease of myocardial respiratory activity were observed in both group B and C at 0.5 PTH, and they became more and more serious afterwards. However, the changes in the above indices in D group were much better than those in B group (P < 0.05 or 0.01), and no difference was found between D (92.8 +/- 4.0)% and C [(100.0 +/- 0.0) %, P > 0.05] groups. CONCLUSION: Hypoxia played a role in the myocardial microtubule damage as well as in the opening of MPTP. Moreover, hypoxia could also impair the mitochondrial respiratory function. Microtubule destabilizing agent could reproduce well the process of hypoxia induced microtubule damage, while the stabilizing agent exerted protective effect by improving the transition of mitochondrial permeability and the mitochondria respiratory function.