Development of a Convolutional Neural Network-Based Colonoscopy Image Assessment Model for Differentiating Crohn's Disease and Ulcerative Colitis.

Objective: Evaluation of the endoscopic features of Crohn's disease (CD) and ulcerative colitis (UC) is the key diagnostic approach in distinguishing these two diseases. However, making diagnostic differentiation of endoscopic images requires precise interpretation by experienced clinicians, which remains a challenge to date. Therefore, this study aimed to establish a convolutional neural network (CNN)-based model to facilitate the diagnostic classification among CD, UC, and healthy controls based on colonoscopy images. Methods: A total of 15,330 eligible colonoscopy images from 217 CD patients, 279 UC patients, and 100 healthy subjects recorded in the endoscopic database of Tongji Hospital were retrospectively collected. After selecting the ResNeXt-101 network, it was trained to classify endoscopic images either as CD, UC, or normal. We assessed its performance by comparing the per-image and per-patient parameters of the classification task with that of the six clinicians of different seniority. Results: In per-image analysis, ResNeXt-101 achieved an overall accuracy of 92.04% for the three-category classification task, which was higher than that of the six clinicians (90.67, 78.33, 86.08, 73.66, 58.30, and 86.21%, respectively). ResNeXt-101 also showed higher differential diagnosis accuracy compared with the best performing clinician (CD 92.39 vs. 91.70%; UC 93.35 vs. 92.39%; normal 98.35 vs. 97.26%). In per-patient analysis, the overall accuracy of the CNN model was 90.91%, compared with 93.94, 78.79, 83.33, 59.09, 56.06, and 90.91% of the clinicians, respectively. Conclusion: The ResNeXt-101 model, established in our study, performed superior to most clinicians in classifying the colonoscopy images as CD, UC, or healthy subjects, suggesting its potential applications in clinical settings.

Development of a conceptual model regarding quality of life in Chinese adult patients with strabismus: a mixed method.

BACKGROUND: Substantial challenges have been reported in China in terms of the large number of adult patients with strabismus and their poor quality of life. Quality of life is a cultural concept that varies according to personal feelings and perceptions, and it is influenced by physical, psychological and social factors. However, to date, there has been no mixed-method research of the quality of life of Chinese adult patients with strabismus, and no conceptual model has been reported. This study aimed to utilize mixed methods to explore the influence of strabismus on health-related quality of life in Chinese adult patients and to develop a conceptual model. METHODS: Thirty adult patients with strabismus from three tertiary hospitals in China participated in the interview. In-depth one-to-one interviews were semi-structured and addressed strabismus-related symptoms and the impacts on the participants' quality of life. Transcripts were analysed to identify themes. A self-designed questionnaire was distributed to 448 patients, 437 of whom returned valid questionnaires. Descriptive statistics and x2 test were conducted. RESULTS: Five themes were revealed regarding the impact of strabismus on patient quality of life: appearance, daily activities, personal development, social interaction, and emotions. In the survey, the top three symptoms (n ≥ 70%) rated by the participants were monocular vision, eye fatigue and physical discomfort. Compared to those without diplopia, the patients who suffered diplopia more often reported experiencing the symptoms of blurred vision, monocular vision, physical discomfort, eye fatigue, cannot estimate depth well and increasing deviation size (all p < 0.05). CONCLUSIONS: This study is the first to examine quality of life among Chinese strabismus patients using both qualitative and quantitative methods and proposing a conceptual model. Symptom burden and appearance were the two original reasons for the decreased quality of life, and they were also the triggers for strabismus patients to visit clinics and undergo surgery. The interventions to treat symptoms burden should be different between patients with and without diplopia.

Rasch Analysis of the Adult Strabismus Quality of Life Questionnaire (AS-20) among Chinese Adult Patients with Strabismus.

BACKGROUND: The impact of strabismus on visual function, self-image, self-esteem, and social interactions decrease health-related quality of life (HRQoL).The purpose of this study was to evaluate and refine the adult strabismus quality of life questionnaire (AS-20) by using Rasch analysis among Chinese adult patients with strabismus. METHODS: We evaluated the fitness of the AS-20 with Rasch model in Chinese population by assessing unidimensionality, infit and outfit, person and item separation index and reliability, response ordering, targeting and differential item functioning (DIF). RESULTS: The overall AS-20 did not demonstrate unidimensional; however, it was achieved separately in the two Rasch-revised subscales: the psychosocial subscale (11 items) and the function subscale (9 items). The features of good targeting, optimal item infit and outfit, and no notable local dependence were found for each of the subscales. The rating scale was appropriate for the psychosocial subscale but a reduction to four response categories was required for the function subscale. No significant DIF were revealed for any demographic and clinical factors (e.g., age, gender, and strabismus types). CONCLUSION: The AS-20 was demonstrated by Rasch analysis to be a rigorous instrument for measuring health-related quality of life in Chinese strabismus patents if some revisions were made regarding the subscale construct and response options.

The effect of concentration and duration of normobaric oxygen in reducing caspase-3 and -9 expression in a rat-model of focal cerebral ischaemia.

The aim of this study was to determine the effect of different concentrations of normobaric oxygen (NBO) on neurological function and the expression of caspase-3 and -9 in a rat model of acute cerebral ischaemia. Sprague-Dawley rats (n=120) were randomly divided into four groups (n=30 per group), including 3 groups given NBO at concentrations of 33%, 45% or 61% and one control group given air (21% oxygen). After 2h of ischaemic occlusion, each group was further subdivided into six subgroups (n=5) during reperfusion according to the duration (3, 6, 12, 24, 48 or 72h) and concentration of NBO (33%, 45% or 61%) or air treatment. The Fluorescence Quantitative polymerase chain reaction (PCR) and immunohistochemistry were used to detect caspase-3 and -9 mRNA and protein relative expression respectively. The Neurologic Impairment Score (NIS) was significantly lower in rats given 61% NBO ≥3h after reperfusion when compared to the control group (P<0.05, Mann-Whitney U). NBO significantly reduced caspase-3 and -9 mRNA and protein expression when compared to the control group at all NBO concentrations and time points (P<0.05, ANOVA). The expression of caspase-3 and -9 was lower in the group given 61% NBO compared any other group, and this difference was statistically significant when compared to the group given 33% NBO for ≥48h and the control group (both P<0.05, ANOVA). These findings indicate that NBO may inhibit the apoptotic pathway by reducing caspase-3 and -9 expression, thereby promoting neurological functional recovery after stroke.

Comparison of the Adult Strabismus Quality of Life Questionnaire (AS-20) with the Amblyopia and Strabismus Questionnaire (ASQE) among adults with strabismus who seek medical care in China.

BACKGROUND: The impact of strabismus on visual function, self-image, self-esteem, and social interactions might decrease health-related quality of life (HRQoL). This study aimed to evaluate the psychometric properties and clinical applications of two strabismus-specific HRQoL questionnaires in the cultural context of China. METHODS: The Chinese versions of the Adult Strabismus Quality of Life Questionnaire (AS-20) and the Amblyopia and Strabismus Questionnaire (ASQE) were self-administered to 304 adults with strabismus. The Cronbach's α coefficient was calculated to assess the internal consistency reliability. The criterion-related validity was identified by exploring Spearman's correlation with the most widely used vision-specific quality of life questionnaire NEI-VFQ-25. One-way ANOVA was employed to examine the differences in the quality of life of strabismus patients with visually normal adults and with other eye diseases patients. RESULTS: Significantly positive correlations with NEI-VFQ-25 were shown in both scales (r = 0.21 - 0.44, p <0.05, p <0.01). Both scales could distinguish individuals with strabismus from visually normal adults (p <0.001) and adults with other eye diseases (p <0.001). The overall Cronbach's α value were 0.91 for the AS-20 and 0.89 for the ASQE; and for the subscales, the α value ranged from 0.68 to 0.90. CONCLUSION: This was the first cross-sectional study to compare the psychometric properties of two strabismus-targeted questionnaires, AS-20 and ASQE in the context of Chinese culture. Both AS-20 and ASQE showed satisfactory and comparable properties for measuring HRQoL in strabismus patients.

Loss of ASAP3 destabilizes cytoskeletal protein ACTG1 to suppress cancer cell migration.

ArfGAP with SH3 domain, ankyrin repeat and PH domain 3 (ASAP3), previously known as ACAP4, DDEFL1 and UPLC1, is considered to be an important regulator in cancer cell migration/invasion and actin-based cytoskeletal remodeling. However, the underlying mechanisms through which ASAP3 mediates these processes are not well-elucidated. This study reported that in certain types of cancer cells, loss of ASAP3 suppressed cell migration/invasion, in part by destabilizing γ-actin-1 (ACTG1), a cytoskeletal protein considered to be an integral component of the cell migratory machinery, essential for the rearrangement of the dynamic cytoskeletal networks and important in diseases, such as brain malformation, hearing loss and cancer development. The data, for the first time, link ASAP3 with ACTG1 in the regulation of cytoskeletal maintenance and cell motility.

Development and application of the Chinese version of the adult strabismus quality of life questionnaire (AS-20): a cross-sectional study.

BACKGROUND: Patients with strabismus experience visual dysfunction, self-image disorders, low self-esteem, and social and emotional barriers, which adversely influence their health-related quality of life (HRQoL). Currently no strabismus-specific questionnaire is available in China to identify patients' quality of life and to evaluate the effectiveness of strabismus treatment. The aims of the present study were to validate the Chinese-language version of the Adult Strabismus Quality of Life Questionnaire (AS-20) and to evaluate the impacts of strabismus on the quality of life among Chinese strabismus patients. METHODS: Two hundred and fifty-five Chinese adults with strabismus, one hundred visually normal adults and one hundred patients with other eye diseases completed the Chinese version of AS-20. Psychometric properties of the Chinese AS-20 were examined by Cronbach's α coefficient, test-retest and split-half reliability, and construct and criterion-related validity. Independent-samples t test and one-way ANOVA analyses were conducted to explore the impact of demographic factors and clinical characteristics on HRQoL in Chinese strabismic adults. RESULTS: The final AS-20 in Chinese (AS-C) included 18 items and two subscales: psychosocial (12 items) and function (6 items). The Cronbach's α was 0.908 for overall scale, with 0.913 and 0.808 for 'psychosocial' and 'function' subscales respectively, indicating high internal consistency reliability. The mean of the overall AS-C score among strabismus patients was 62.80 ± 18.94, significantly lower than that in visually normal adults (t = -18.693, P < 0.001), and in patients with other eye diseases (t = -5.512, P < 0.001). CONCLUSIONS: The AS-C is a culturally appropriate tool to evaluate the HRQoL in Chinese strabismus adults. The psychosocial health well-being and overall quality of life in strabismic patients should receive greater emphasis.

Whole-genome sequencing reveals untapped genetic potential in Africa's indigenous cereal crop sorghum.

Sorghum is a food and feed cereal crop adapted to heat and drought and a staple for 500 million of the world's poorest people. Its small diploid genome and phenotypic diversity make it an ideal C4 grass model as a complement to C3 rice. Here we present high coverage (16-45 × ) resequenced genomes of 44 sorghum lines representing the primary gene pool and spanning dimensions of geographic origin, end-use and taxonomic group. We also report the first resequenced genome of S. propinquum, identifying 8 M high-quality SNPs, 1.9 M indels and specific gene loss and gain events in S. bicolor. We observe strong racial structure and a complex domestication history involving at least two distinct domestication events. These assembled genomes enable the leveraging of existing cereal functional genomics data against the novel diversity available in sorghum, providing an unmatched resource for the genetic improvement of sorghum and other grass species.

[Advances in the research of fluid resuscitation and its monitoring in burn patients].

Shock is the one of the most serious complications during the early stage of burn injury. Early effective fluid resuscitation, enabling the burn patient to pass through the shock stage smoothly and uneventfully, plays a necessary and essential role in the prevention of the subsequent organ complications, reduction of mortality and morbidity, and improvement in life quality. Rapid restoration of blood volume is the fundamental measure to prevent burn shock. In this review, the history and the current status of several important issues related to burn shock resuscitation, including the fluid replacement formula, quality of fluids, and monitoring of physiological parameters, were over viewed. The authors also proposed that a new therapeutic strategy to prevent microvascular permeability should be emphasized and developed in future, which may hopefully act as the most basic approach to prevent burn shock and its related complications.

A Disaster Response and Management Competency Mapping of Community Nurses in China.

BACKGROUND: It is widely accepted in many parts of the world thatcommunity nurses are of vital importance in various phases of disaster response and management. In China, however, it is not clear whether the Chinese community nurses are able to assume disaster-related duties due to the lack of a systematic assessment. METHODS: A pre-designed and well-tested questionnaire was employed to evaluate the competency in disaster response and management among 205 valid registered Chinese community nurses between September and October 2009. Statistical analyses were performed with SPSS Version 13.0 using one way ANOVA, Least Significant Difference (LSD) and multiple stepwise regression analysis. RESULTS: This group of Chinese community nurses scored at an intermediate level of competency (a score of 3.68 (SD 0.48) out of a perfect score of 5) in disaster response and management, suggesting that they have the basic ability to participate in disaster-related nursing. Four factors, namely, Experiences in Disaster Relief, Participation in Disaster Training, the Age and Duration in Job, were identified to be the predominant factors contributing significantly to the integrated competency in disaster response and management of an individual. CONCLUSION: Most of the Chinese community nurses have basic qualifications and competencies to undertake the responsibilities of disaster response and management. However, more targeted disaster training including virtual-reality based drills should be provided in order to improve their competency.

[Influence of histatin 1 on the proliferation and migration of HaCaT cells].

OBJECTIVE: To study the influence of histatin 1 (Hst1) on the proliferation and migration of human epidermal cell line HaCaT. METHODS: (1) HaCaT cells were routinely cultured and divided into control group, 100, 30, and 3 µg/mL Hst1 groups, 10 ng/mL recombinant human epidermal growth factor (rhEGF) group, and 30 µg/mL Hst1 + 10 ng/mL rhEGF group, according to the random number table (the same dividing method used for following grouping), with 27 samples in each group. NO stimulating factor was added in control group, while Hst1 and(or) rhEGF in corresponding concentration(s) was (were) added in the latter 5 groups. Cell proliferation was assayed by cell counting method at post culture hour (PCH) 24, 48, and 72. (2) HaCaT cells were divided into control group and 100, 30, and 3 µg/mL Hst1 groups, with 27 samples in each group. NO stimulating factor was added in control group, while Hst1 in corresponding concentration was added in the latter 3 groups. Cell cycle was assayed with flow cytometry at PCH 24, 48, and 72, and PI was calculated. (3) HaCaT cells were divided into control group, 30 µg/mL Hst1 group, 10 ng/mL rhEGF group, 30 µg/mL Hst1 + 10 ng/mL rhEGF group, 15 µg/mL Hst1 + 5 ng/mL rhEGF group, and 15 µg/mL Hst1 + 10 ng/mL rhEGF group, with 10 samples in each group. NO stimulating factor was added in control group, while Hst1 and(or) rhEGF in corresponding concentration(s) was (were) added in the latter 5 groups. Cells in each group were divided into two portions: cells in one portion were treated by mitomycin C for 2 hours, while cells in the other portion were not. Scratching assay was conducted in both portions of cells. Cell migration was measured at post scratching hour (PSH) 0, 16, and 24, and the wound-area healing rate was calculated. Data were processed with analysis of variance, and LSD- t test or Dunnett t test was applied in paired comparison among groups. RESULTS: (1) At PCH 24, the cell numbers in 10 ng/mL rhEGF group and 30 µg/mL Hst1 + 10 ng/mL rhEGF group were significantly higher than that in control group (with t values respectively 3.813, 5.410, P < 0.05 or P < 0.01). Except for cell numbers in 30 µg/mL Hst1 group and 3 µg/mL Hst1 group at PCH 48, cell numbers in the other groups as treated by Hst1 and (or) rhEGF were significantly higher than those in control group at PCH 48 and 72 (with t values from 7.754 to 24.979, P values all below 0.01). At PCH 72, the cell number was obviously higher in 100 µg/mL Hst1 group [(19.21 ± 0.59)×104] than in 30 µg/mL Hst1 group [(16.19 ± 0.53)×104)] and 3 µg/mL Hst1 group [(15.38 ± 0.13)×104], with t values respectively 11.391, 19.017, P values all below 0.01. The cell number was higher in 30 µg/mL Hst1 + 10 ng/mL rhEGF group than in 30 µg/mL Hst1 group, 3 µg/mL Hst1 group, and 10 ng/mL rhEGF group (with t values from 4.579 to 34.884, P < 0.05 or P < 0.01). Cell numbers in all groups increased with prolongation of time. (2) Compared with those in control group at PCH 24 and 48, the percentage of cells in G0/G1 phase was decreased, the percentage of cells in S phase was increased (except for cell percentage of 30 µg/mL Hst1 group at PCH 24), and PI value was significantly increased in 100 µg/mL Hst1 group and 30 µg/mL Hst1 group (with t values from 4.752 to 16.104, P values all below 0.01). The PI value in 3 µg/mL Hst1 group was obviously higher than that in control group only at PCH 48 (t = 4.609, P < 0.01). At PCH 72, only the PI value in 100 µg/mL Hst1 group was higher than that in control group (t = 8.005, P < 0.01). Compared among the groups treated by Hst1, the percentage of cells in G0/G1 phase showed an elevating trend, and the percentage of cells in S phase and the PI value showed a declining trend along with the decrease in Hst1 concentration at each time point. Compared within each group treated by Hst1, the percentage of cells in G0/G1 phase declined first and then elevated, while the percentage of cells in S phase and the PI value elevated first and then declined along with prolongation of time. (3) Without treatment of mitomycin C, the wound-area healing rate in 30 µg/mL Hst1 group (75.9 ± 3.9)% at PSH 16 was significantly higher than those in control group and 10 ng/mL rhEGF group [(53.0 ± 3.5)%, (61.7 ± 2.5)%, with t values respectively 12.241, 7.598, P values all below 0.01], but lower than those in 30 µg/mL Hst1 + 10 ng/mL rhEGF group, 15 µg/mL Hst1 + 5 ng/mL rhEGF group, and 15 µg/mL Hst1 + 10 ng/mL rhEGF group [(95.0 ± 4.1)%, (97.0 ± 3.7)%, (80.5 ± 5.9)%, with t values from -11.324 to -2.502, P < 0.05 or P < 0.01]. After being treated by mitomycin C, the wound-area healing rate in 30 µg/mL Hst1 group at PSH 16 [(54.1 ± 4.5)%] was higher than that in control group [(35.8 ± 5.7)%, t = 7.790, P < 0.01], but lower than that in the same Hst1 concentration but without mitomycin C treatment group (t = -10.863, P < 0.01). There was no statistically significant difference in the wound-area healing rate between 30 µg/mL Hst1 group and other groups treated by Hst1 and rhEGF at PSH 16 (with t values from 0.061 to 2.030, P values all above 0.05). Compared within each group with or without treatment of mitomycin C, the wound-area healing rate at PSH 16 was not significantly different from that at PSH 24 (with F values from 0.856 to 3.062, P values all above 0.05). CONCLUSIONS: Hst1 can promote the proliferation and migration of HaCaT cells. It has synergic effect with rhEGF on the promotion of cell proliferation, but their synergic effect on cell migration is not obvious.

Reduced CD27 expression on antigen-specific CD4+ T cells correlates with persistent active tuberculosis.

OBJECTIVE: CD27, a member of the tumor necrosis factor receptor family, has important role in generation of T cell immunity. In this study, association of CD27 expression on mycobacterial antigen-specific CD4+ T cells with pulmonary tuberculosis (TB) was investigated. METHODS: Mycobacterial antigen-specific CD4+ T cells were identified based on CD154 expression and CD27 expression on antigen-specific CD4 T cells was analyzed by flow cytometry. RESULTS: Compared with tuberculin-positive controls, patients with bacterial culture-positive pulmonary TB had significantly reduced CD27 expression on antigen-specific CD4 T cells. The persistent active TB patients had much lower percentages of CD27+ antigen-specific CD4 T cells than culture-positive new TB patients (P=0.008) and healthy controls (P=0.005). Logistic regression analysis on frequencies of CD27-expressing antigen-specific CD4 T cells and TB patients' clinical characteristics indicated that low percentage of CD27+ antigen-specific CD4 T cells correlated significantly with persistent active tuberculosis (P=0.002, odds ratio=19.6). CONCLUSION: It is concluded that frequency of CD27+ antigen-specific CD4 T cells could be used as an immunological marker for persistent active TB.

[Expression of B7-H1 protein in human pancreatic carcinoma tissues and its clinical significance].

BACKGROUND AND OBJECTIVE: B7-H1, a member of B7 family, is expressed in tumor cells and has emerged as an important immune modulator capable of suppressing host immunity by inhibiting T cells function. This study was to probe into the correlation between the expression level of B7-H1 protein in pancreatic carcinoma tissues and clinicopathological characteristics and prognosis. METHODS: The expression of B7-H1 was measured in 40 cases of pancreatic carcinoma tissues and 10 cases of normal corresponding paracarcinoma tissues by immunohistochemistry. The relationship between the expression level of B7-H1 and clinicopathological characteristics and survival was analyzed. RESULTS: The positive rate of B7-H1 was significantly higher in the tumor tissues [45.00% (18/40)] than in the normal corresponding paracarcinoma tissues [0(0/10)] (P<0.05); moreover, B7-H1 expression was significantly associated with the staging of tumor and preoperative serum CA19-9 level (P<0.05). The multivariate cox proportional hazards regression analysis of prognostic factors for overall survival and relapse-free survival showed that the expression of B7-H1 was an independent factor for poor prognosis. CONCLUSION: B7-H1 protein was expressed in human pancreatic carcinoma tissues, and was associated with the prognosis.

[Expression and significance of membrane skeleton protein 4.1 family in non-small cell lung cancer].

BACKGROUND AND OBJECTIVE: Protein 4.1, a component of cell membrane skeleton, plays a role in maintaining the shape and mechanical stability of erythrocytes. Recent researches showed that protein 4.1 may be associated with the development of tumors. This study was to investigate the expression and significance of membrane skeleton protein 4.1 family members (4.1B, 4.1R, 4.1N and 4.1G) in human non-small cell lung cancer (NSCLC). METHODS: The expression of proteins 4.1B, 4.1R, 4.1N and 4.1G in 147 specimens of NSCLC was detected by EnVision plus immunohistochemistry. The correlations of 4.1B, 4.1R, 4.1N and 4.1G expression to clinicopathologic features of NSCLC were analyzed by Wilcoxon rank sum test and Spearman rank correlation analysis. RESULTS: The protein levels of 4.1B, 4.1R and 4.1N were significantly lower in lung squamous cell carcinoma tissues than in adjacent normal tissues (P<0.01). The protein levels of 4.1B, 4.1R, 4.1N and 4.1G were significantly lower in lung adenocarcinoma tissues than in adjacent normal tissues (P<0.05). The protein levels of 4.1B and 4.1G were significantly lower in lung squamous cell carcinoma tissues than in lung adenocarcinoma tissues (P<0.05). Protein 4.1G expression in squamous cell carcinoma was positively correlated to tumor cell differentiation (rs=0.386,P<0.01). In adenocarcinoma, the expression of proteins 4.1B, 4.1N and 4.1G were positively correlated to tumor cell differentiation (rs=0.276, P<0.05; rs=0.248,P<0.05; rs=0.268, P <0.05). The expression of protein 4.1s in squamous cell carcinoma and adenocarcinoma were not related to lymph node metastasis, tumor size, patients'age and sex (P>0.05). CONCLUSIONS: Protein 4.1s are weakly expressed in NSCLC tissues than in adjacent normal tissues. The expression of proteins 4.1B, 4.1N and 4.1G are related to tumor cell differentiation.

Identification of a new broad-spectrum CD8+ T cell epitope from over-expressed antigen COX-2 in esophageal carcinoma.

Cyclooxygenase-2 (COX-2) has been found to be over-expressed in esophageal carcinoma (EC) and it could be considered as a potential tumor-associated antigen (TAA). In the present study, six candidate peptides from COX-2 were firstly predicted and synthesized. Among them, P(479) had the highest affinity and stability toward both HLA-A *0201 and HLA-A *03 molecules and it could significantly promote the IFN-gamma release. The cytotoxic T lymphocytes (CTLs) induced by P(479) could specifically lyse COX-2-expressed EC cell lines, EC-1 (HLA-A3 supertype) and EC-9706 (HLA-A2 supertype). These results suggested that P(479) as a novel broad-spectrum T cell epitope would be very useful in immunotherapy against esophageal carcinoma.

[Clone, expression and identification of penicillin binding protein 2a of methicillin-resistant Staphylococcus aureus isolated from patients].

OBJECTIVE: To clone, express and identify the mecA fragment which encoded penicillin binding protein 2a (PBP2a) from methicillin-resistant staphylococcus aureus (MRSA) isolated from patients by gene recombination method. METHODS: According to the sequence of mecA gene recorded in GenBank, the primer of mecA fragment which encoded amino acids 25 - 668 of PBP2a was designed. Then the mecA fragment was amplified by PCR and cloned into pQE30 plasmid. After being identified by enzyme digestion and sequencing, the recombinant plasmid was transferred into E. coli M15 [pREP4], and then its expression was induced by 1 mmol/L Isopropy-beta-D-Thiogalactoside (IPTG). The expression product was analyzed by SDS-PAGE, protein sequencing and mass spectroscopy. RESULTS: The recombinant pQE30- mecA had been successfully constructed. The result of sequencing showed that the mecA fragment had 1932 bases, including 9 bases undergoing mutation. After being induced for 6 hours by IPTG, the soluble protein in M15 (pQE30- mecA), with a relative molecular weight of 74 x 10(3), was found by SDS-PAGE. The soluble protein had been confirmed to be PBP2a after identification. CONCLUSION: The soluble PBP2a of MRSA isolated from patients is expressed successfully by gene recombinant technology.

Effects of preoperatively selected gut decontamination on cardiopulmonary bypass-induced endotoxemia.

OBJECTIVE: To evaluate the influence of preoperatively selected gut decontamination (SGD) on intestinally derived endotoxemia(ETM) in patients with rheumatic heart disease undergoing valve replacement operation with cardiopulmonary bypass(CPB). METHODS: Thirty patients were randomly divided into control group and SGD group. The patients in control group underwent preoperative bowel preparation, i.e, diet preparation and enema. The patients in SGD group were administrated 100 mg Tobramycin, 40 mg garlicin and 20% Lactulose for 10 ml three times per day for 3 days besides routinely preoperative bowel preparation. Bacteria cultivation and identification and Gram staining of feces in both groups were used to evaluate species of intestinal flora and their ratios. The levels of endotoxin, D-lactate, TNF-alpha and complement 3 were determined at four time points of anesthetic induction, CPB end, 2 h after CPB, 24 h after CPB. And the related clinical biochemical and clinical markers were recorded. RESULTS: Aerobic gram-negative bacilli (AGNB) ratio in post-SGD group decreased significantly as compared with that in control group and pre-SGD group (P less than 0.05). The level of D-lactate reduced significantly at time points of anesthetic induction and 2 h after CPB (P less than 0.05). Endotoxin levels of patients in both groups elevated significantly after CPB (P less than 0.05), and endotoxin levels of the patients in SGD group decreased significantly at points of CPB end (P less than 0.01) and 24 h after CPB (P less than 0.05) compared with those in control group. The levels of TNF-alpha and complement 3 were similar in both groups as well as clinical and biochemical markers. CONCLUSIONS: CPB induces endotoxemia, while the regime of SGD is an effective way to prevent endotoxemia but may not affect activation of inflammatory media and clinical outcomes.

[Effects of soybean isoflavones on the cell cycles, the cell apoptosis and the proliferation of spleen in radiated mice].

AIM: To study effect of soybean isoflavones (SI) on spleen in radiated mice. METHODS: 90 male mice were randomly divided into control group, radiated group, radiated plus 0.5% dose SI group. After 2-week feeding, the mice received 4.0 Gy 137Cs gamma-radiation, the cell cycles, cell apoptosis and proliferation on the spleen and the spleen index were observed in radiated after 12 h, 24 h, 1 week and 2 weeks. RESULTS: After the mice were radiated, the spleen were significantly atrophy, the rate of the cell apoptosis and the cell cycles of G0-G1 phase in splenocytes were significantly increased (P < 0.01), the cell cycles rate of S phase and the proliferation index were significantly decreased in spleen (P < 0.05). Compared with radiated group, the spleen atrophy and the rate of the cell cycles of G0-G1 phase were significantly decreased (P < 0.05), and the cell cycles of G2-M phase and the proliferation index were significantly increased (P < 0.05) in the mice supplied 0.5% soybean isoflavones. CONCLUSION: The soybean isoflavones could significantly increase spleen radioprotective effect in mice.

[Effect of oxygen therapy for injury of intestinal mucosal barrier of rabbits in high altitude hemorrhagic shock].

OBJECTIVE: To investigate the protective effects of oxygen therapy on gut barrier function of rabbits after hemorrhagic shock at high altitude. METHODS: Fifteen rabbits were exposed to a simulated 4000 m high altitude in a hypobaric chamber, and they were randomly divided into two groups of treatment and control. Then hemorrhagic shock was induced through bleeding to mean artery pressure 40 mmHg (1 mmHg=0.133 kPa). After 1 hour, the rabbits were resuscitated with infusion of total volume of shed blood and balanced saline equivalent to 1.5 times of the shed blood. In addition, the animals of the treatment group were given oxygen therapy. RESULTS: After 4 hours treatment, the mean arterial pressure of the treatment group was increased significantly in comparison with the control group, and the concentrations of plasma lactate in the former were significantly decreased after treatment for 4 hours and 8 hours. The plasma lipopolysaccharide levels in the treatment group were significantly lower than those in the control after treatment, and the histologic evidence of damage to the intestinal mucosa of the rabbits in the treatment group was less marked than that in the control group (P<0.05 or P<0.01). CONCLUSION: Hemorrhagic shock at high altitude may cause disruption of the intestinal mucosa barrier and oxygen therapy could afford protection to the intestinal mucosa barrier in such circumstance.

[Effects of genistein on bone mineralization in ovariectomized rats].

AIM: To investigate the effects of genistein on bone mineralization in ovariectomized rats. METHODS: Forty-seven Wistar rats were randomly allocated into six groups: sham-operated (sham), ovariectomized (ovx), ovariectomized supplied with diethyl stilbestrol (E, 20 microg x kg bw(-1) x d(-1)) or genistein (25, 50, 100 mg x kg bw(-1) x d(-1)). After the rats had been fed for three months, analysis of the bone mineral density, parameters related to mineralization, bone content of Ca, P, Mg, Mn and Zn and serum concentration of parathyroid calcitonin and estrogen was performed. RESULTS: Bone mineral density, bone Ca, P, Zn and Mg content and serum estrogen concentration in ovariectomized rats were significantly decreased, but mean osteoid width increased, mineralization lag time and osteoid maturation period prolonged compared with sham animals. After three months supplementation to ovariectomized rats, bone Ca, P and Mg content increased, mean osteoid width decreased, mineralization lag time and osteoid maturation period shortened compared with ovariectomized animals. CONCLUSION: Genistein promotes bone mineralization by increasing bone Ca, P, Mg and adjusting serum calcitonin to prevent osteoporosis.