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The aim of this study is to establish a multiplex polymerase chain raction (PCR) to identify of four kinds of laboratory animal pathogens: Pasteurella multocida, Bordetella bronchiseptica, Mycoplasma pneumoniae and Klebsiella pneumoniae.Methods Specific primers were designed based on GenBank data.The multiplex PCR system was established through optimization of multiple PCR and detection of its specificity and sensitivity.This technique was used to test artificially infected samples and tracheal secretions of experimental animals (rat, mouse, guinea pig, rabbit, hamster), and comparing the detection results by this method and traditional detection test.Results Target bands of Pasteurella multocida (356 bp), Bordetella bronchiseptica (237 bp), Mycoplasma pneumoniae (266 bp), and Klebsiella pneumoniae (142 bp) were obtained, with a detection sensitivity of Klebsiella pneumoniae of 10 pg, and that of Pasteurella multocida, Bordetella bronchiseptica and Mycoplasma pneumoniae of 1 pg by this newly developed multiplex PCR assay.No target bands were observed from the non-specific pathogens of artificially infected samples.The tracheal secretions taken from 45 experimental animals (mice and rabbits) were tested with this new PCR assay, among which 15 cases of Klebsiella pneumonia and 9 cases of Pasteurella multocida were detected as positive, while all the results of traditional method and serological test were negative.Conclusions A simple, rapid, specific and highly sensitive multiplex PCR system has been successfully established.It is valuable for detection of Pasteurella multocida, Bordetella bronchiseptica, Mycoplasma pneumoniae, and Klebsiella pneumoniae in laboratory animals.
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BACKGROUND:The low oxygen environment after femoral fracture and cerebral trauma wil induce series of related cytokines expression, including hypoxia-inducible factor 1αand core binding factorα1, which play key roles in regulating bone healing. However, whether the accelerated bone healing is correlated with the expression of hypoxia-inducible factor 1αand core binding factorα1 is stil unknown. OBJECTIVE:To construct rat models of brain injury, to compare the expression level of hypoxia-inducible factor 1αand core binding factorα1 in femoral fracture combined with cerebral trauma rats and simple femoral fracture rats, and to assess the influence of cerebral trauma on bone healing. METHODS:Rats were randomly divided into blank group, simple femoral fracture group and femoral fracture combined with cerebral trauma group. At 1, 2, 3 and 5 weeks after modeling, rats were executed. Bone healing was evaluated using femoral fracture end X-ray score and hematoxylin and eosin staining at cal us tissues. Besides, the expression levels of hypoxia-inducible factor 1αand core binding factorα1 of three groups were determined with immunohistochemistry. RESULTS AND CONCLUSION:Bone healing in the femoral fracture combined with cerebral trauma group was better than that of simple femoral fracture group. There was significant difference in the expression level of hypoxia-inducible factor 1αand core binding factorα1 between the simple femoral fracture group and femoral fracture combined with cerebral trauma group (P<0.05). At the same time, the level of simple femoral fracture group and femoral fracture combined with cerebral trauma group was significantly higher than that of blank group, and that in femoral fracture combined with cerebral trauma group was significantly higher than that of simple femoral fracture group (P<0.05). Results verified that the expression levels of hypoxia-inducible factor 1αand core binding factorα1 of rats with femoral fracture combined with cerebral trauma were significantly high, which may be the major reason why the bone healing was accelerated after fracture combined with brain injury.
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Objective To investigate the influence of apolipoprotein E gene (APOE) polymorphism on the acute-phase brain electrical activity after mild/moderate traumatic brain injury. Methods The clinical data of 112 patients with mild/moderate traumatic brain injury were collected and the APOE genotypes were identified by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). The brain electrical activity in every patient was recorded twice by using electroencephalogram within one week after injury. Qualitative and quantitative methods were used to determine the variations of brain electrical activity. Chi-square test, variance analysis and logistic regression analyses via SPSS version 11.5 were performed among APOE genotypes, electroencephalogram data and clinical data. Results The distributions of APOE genetypes and alleles matched Haldy-Weinberg Law in 112 patients. Of 22 patients with APOEε4, 12 patients (55%) presented with deteriorated electroencephalogram, which was significantly higher than those (16 of 90 patients, 18%) without APOEε4 (P ＜ 0. 01). Comparison of the first and second electroencephalograms demonstrated that the slow waves were increased significantly in patients with APOEε4 ( P ＜ 0. 01 ) but decreased in patients with APOEε2 and APOEε3 (P ＜0.05). The reduction of slow waves in APOEε2 carriers was more obvious than APOEε3 carriers (P ＜0.05). Univariate and multivariate logistic regression analyses showed that APOEε4 was a risk factor to electroencephalogram aggravation after traumatic brain injury. Conclusion APOEε4 is a risk factor to electroencephalogram aggravation during acute stage after mild/moderate traumatic brain injury. However,APOEε2 seems to be beneficial for recovery of brain electrical activity.
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Objective To discuss the psychological behavior intervention methods on reducing the violence behavior of schizophrenia patients. Methods 120 schizophrenia patients were admitted from January to December, 2007 and randomly assigned into the behavior intervention group (group A), the psy-chological support group (group B), the health education group (group C) and the control group (group D) with 30 cases in each group. Group A received routine nursing plus behavior intervention, group B was giv-en routine nursing plus psychological support, group C adopted routine nursing plus health education, group D only received routine nursing. The intervention effect was evaluated with MOAS and the relapse rate of violence behavior went through statistics. Results After intervention, the scores of MOAS decreased sig-nificantly in group A, B, C and D compared with those before intervention. The reduction of MOAS scores in group A, B and C was significantly higher than that of group D, with the highest reduction score in group A and group B and C followed subsequently. The relapse rates of the violence behavior in group A, B and C were significantly lower than those of group D. Conclusions Routine nursing, health education, psycho-logical support and behavior intervention can all reduce and decrease the violence behavior and the inci-dence of the violence behavior, among which behavior intervention proves to be the most effective, and psy-chological support and health education go subsequently.
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Objective To determine the relationship between polymorphism of apolipoprotein E gene (APOE) and electroencephlogram in patients with mild/moderate traumatic brain injury. Methods (1) Venous blood for 2 ml was collected from 81 patients with mild/moderate traumatic brain injury on admission. APOE genotype was identified by PCR restriction fragment length polymorphism ( PCRRFLP). (2) All the patients were monitored by electroencephalogram for 2-3 times within a week after injury. X2 test and logistic regression analysis via SAS version 8.2 were performed to analyze the results of genotype and electroencephalogram and clinical data. Results The distributions of genetypes and alleles among 81 patients matched with Haldy-Weinberg Law. The findings of electroencephalogram were significantly different between patients with and without APOEε4 (P<0.05). Ten (63%) out of 16 patients with APOEε4 showed an aggravated electroencephalogram,while only 16 (25%) out of 65 patients without APOEε4 showed the same results of electroencephalogram. Logistic regression analyses showed that APOEε4 was a risk factor for electroencephalogram aggravation after traumatic brain injury. Conclusion APOEε4 is a risk factor for electroencephalogram aggravation during acute stage after mild/moderate traumatic brain injury.
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BACKGROUND: Applying naloxone in acute brain injury can sustain the cerebral perfusion pressure(CPP), alleviate the cerebral edema and prevent the secondary brain damage to a certain degree. But the dosage and the administration of naloxone in clinical practices vary substantially according to the literatures.OBJECTIVE: To investigate the effect of different doses of naloxone on the changes in the absolute power values of electroencephalography(EEG) in acute brain injury, and study the protective effects of naloxone at different doses.DESIGN: Case-control study based on patients.SETTING: Neurosugery department of a hospital affiliated to a university PARTICIPANTS: From January 2002 to April 2003, at the Intensive Care Unit(ICU) of theNeurosugery Department of the First Hospital Affiliated to the Chongqin Medical University, 86 patients with moderate or severe acute closed brain injury were selected. Of all the patients, 59 were male and 27 were female, aged between 18 - 65.METHODS: According to the degree of injury graded by Glasgow Coma Scale(GCS), the 86 patients bearing acute brain injury were divided into 3 groups: GCS 3 - 5 group, GCS 6 - 8 group and GCS 9 - 12 group. Each group contained a naloxone treatment group and a matched control group. The naloxone treatment group consisted of a low-dose naloxone subgroup and a large-dose naloxone subgroup. The changes in the total power value of EEG before treatment and at the time of 30 minutes, 1, 2, 24, 48, 72 and 120 hours after treatment were measured respectively using quantitative EEG monitor.MAIN OUTCOME MEASURES: The changes in the total power value of the patients' EEG before and after treatment were observed and recorded.RESULTS: The difference between the total power of EEG of the GCS 9 - 12naloxone treatment group 1 hour after a naloxone treatment and that of the matched control group was statistically significant(P ＜ 0.05); The same comparison between the low-dose and the large-dose naloxone subgroups within the GCS 9 - 12 naloxone treatment group yielded no significant difference. In the GCS 6 - 8 naloxone treatment group, the difference between the total power of EEG 1 hour after a naloxone treatment and that of the matched control group was statistically significant, and the large dose subgroup was more significant than the low-dose group. In the GCS 3 - 5 naloxone treatment group, no significant difference between the total power of EEG of the naloxone group and that of the control group could be observed.CONCLUSION: The low-dose naloxone treatment is helpful enough on the intervention for moderate brain injury, and the large-dose naloxone treatment is better than the low-dose on severe brain injury. For the patients with exceptionally severe brain injury, both the two treatments are proved to have no therapeutic effects.
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0.05).Conclusion The variety of ECoG on patients with intracranial tumors undergone surgery is helpful to prognosticate postoperative early epilepsy.
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Objective To explore the effect of Chinese herbal medicine formula "tonifying kidney plus strengthening vital energy" on regulating CD40, CD40L protein and mRNA expression in spleen cell in mice with SHENXU. Methods The interaction of CD40L expressed by activated T cells with CD40 on monocytes, macrophages, or B cells is essential for the production of IL-12 and other cytokines. An experimental model of mice with SHENXU by corticosterone was established to observe the effect of Chinese formula on CD40, CD40L protein and mRNA expression in spleen cells or in ConA stimulated spleen cells by immunohistochemistry and RT-PCR. Results No significant differences were found among 5 groups of mice about the CD40 and CD40L protein expression in spleen cells. Both CD40 and CD40L expression detected by immunohistochemistry decreased seriously in ConA induced cells from SHENXU mice (18.9%?3.3% vs 15.8%?2.3%) as compared to that of normal controls(29.7%?4.2% vs 23.3%?1.3%), and so did the mRNA expression on ConA stimulated cells from SHENXU mice. They were restored in different degrees when the SHENXU mice were cured by different Chinese medicine. The effect of "tonifying kidney plus strengthening vital energy" was stronger than that of "tonifying kidney" and "strengthening vital energy" alone. Conclusions "Tonifying kidney plus strengthening vital energy" enhanced the expression of CD40, CD40L mRNA and protein, and this may be one of the mechanisms of improving the status of immune inhibition resulted by Corticosterone.