Association between PT, PT-INR, and in-hospital mortality in critically ill patients with tumors: A retrospective cohort study.

Objectives: Prothrombin time (PT) and PT-INR are independent predictors of mortality in patients with cancer. The PT and PT-INR of cancer patients are independent predictive variables of mortality. However, whether the PT or PT-INR is related to in-hospital mortality in severely ill patients with tumors remains unknown. Design: This was a case-control study based on a multicenter public database. Settings: This study is a secondary analysis of data extracted from 2014 to 2015 from the Electronic Intensive Care Unit Collaborative Research Database. Participants: The data relevant to seriously ill patients with tumors were obtained from 208 hospitals spread throughout the USA. This research included a total of 200,859 participants. After the samples were screened for patients with combination malignancies and prolonged PT-INR or PT, the remaining 1745 and 1764 participants, respectively, were included in the final data analysis. Primary and secondary outcome measures: The key evaluation methodology was the PT count and PT-INR, and the main outcome was the in-hospital mortality rate. Results: After controlling for confounding variables, we found a curvilinear connection between PT-INR and in-hospital mortality (p < 0.001), and the inflection point was 2.5. When PT-INR was less than 2.5, an increase in PT-INR was positively associated with in-hospital mortality (OR 1.62, 95% CI 1.24 to 2.13), whereas when PT-INR was greater than 2.5, in-hospital mortality was relatively stable and higher than the baseline before the inflection point. Similarly, our study indicated that the PT exhibited a curvilinear connection with in-hospital mortality. On the left side of the inflection point (PT <22), a rise in the PT was positively linked with in-hospital mortality (OR 1.08, 95% CI 1.04 to 1.13, p < 0.001). On the right side of the inflection point, the baseline PT was above 22, and the in-hospital mortality was stable and higher than the PT count in the prior range (OR 1.01, 95% CI 0.97 to 1.04, 0.7056). Conclusion: Our findings revealed that there is a curved rather than a linear link between the PT or PT-INR and in-hospital mortality in critically ill cancer patients. When these two laboratory results are below the inflection point, comprehensive therapy should be employed to reduce the count; when these two laboratory results are above the inflection point, every effort should be made to reduce the numerical value to a value below the inflection point.

Nicotinamide phosphoribosyltransferase as a biomarker for the diagnosis of infectious pleural effusions.

Nicotinamide phosphoribosyltransferase (NAMPT) has been reported to be involved in infectious diseases, but it is unknown whether it plays a role in infectious pleural effusions (IPEs). We observed the levels of NAMPT in pleural effusions of different etiologies and investigated the clinical value of NAMPT in the differential diagnosis of infectious pleural effusions. A total of 111 patients with pleural effusion were enrolled in the study, including 25 parapneumonic effusions (PPEs) (17 uncomplicated PPEs, 3 complicated PPEs, and 5 empyemas), 30 tuberculous pleural effusions (TPEs), 36 malignant pleural effusions (MPEs), and 20 transudative effusions. Pleural fluid NAMPT levels were highest in the patients with empyemas [575.4 (457.7, 649.3) ng/ml], followed by those with complicated PPEs [113.5 (103.5, 155.29) ng/ml], uncomplicated PPEs [24.9 (20.2, 46.7) ng/ml] and TPEs [88 (19.4, 182.6) ng/ml], and lower in patients with MPEs [11.5 (6.5, 18.4) ng/ml] and transudative effusions [4.3 (2.6, 5.1) ng/ml]. Pleural fluid NAMPT levels were significantly higher in PPEs (P < 0.001) or TPEs (P < 0.001) than in MPEs. Moreover, Pleural fluid NAMPT levels were positively correlated with the neutrophil percentage and lactate dehydrogenase (LDH) levels and inversely correlated with glucose levels in both PPEs and TPEs, indicating that NAMPT was implicated in the neutrophil-associated inflammatory response in infectious pleural effusion. Further, multivariate logistic regression analysis showed pleural fluid NAMPT was a significant predictor distinguishing PPEs from MPEs [odds ratio (OR) 1.180, 95% confidence interval (CI) 1.052-1.324, P = 0.005]. Receiver-operating characteristic (ROC) analysis demonstrated that NAMPT was a promising diagnostic factor for the diagnosis of infectious effusions, with the areas under the curve for pleural fluid NAMPT distinguishing PPEs from MPEs, TPEs from MPEs, and IPEs (PPEs and TPEs) from NIPEs were 0.92, 0.85, and 0.88, respectively. In conclusion, pleural fluid NAMPT could be used as a biomarker for the diagnosis of infectious pleural effusions.

Nonalcoholic Fatty Liver Disease-Associated Liver Fibrosis Is Linked with the Severity of Coronary Artery Disease Mediated by Systemic Inflammation.

METHODS AND RESULTS: We conducted a retrospective study of 531 patients with ultrasonogram-confirmed NAFLD who underwent percutaneous coronary intervention (PCI). Then, all patients were separated into four categories by Gensini score (0, 0-9, 9-48, and ≥48) for use in ordinal logistic regression analysis to determine whether NAFLD fibrosis was associated with increased Gensini scores. Mediation analysis was used to investigate whether systemic inflammation is a mediating factor in the association between NAFLD fibrosis and CAD severity. FIB - 4 > 2.67 (OR = 5.67, 95% CI 2.59-12.38) and APRI > 1.5 (OR = 14.8, 95% CI 3.24-67.60) remained to be independent risk factors for the severity of CAD after adjusting for conventional risk factors, whereas among the inflammation markers, only neutrophils and neutrophil-to-lymphocyte ratio (NLR) were independently associated with CAD. Multivariable ordinal regression analysis suggested that increasing Gensini score (0, 0-9, 9-48, and ≥48) was associated with advanced NAFLD fibrosis. ROC curve showed that either fibrosis markers or inflammation markers, integrating with traditional risk factors, could increase the predictive capacity for determining CAD. Inflammation markers, especially neutrophils and NLR, were mediators of the relationship between NAFLD fibrosis and CAD severity. CONCLUSIONS: NAFLD patients with advanced fibrosis are at a high risk of severe coronary artery stenosis, and inflammation might mediate the association between NAFLD fibrosis and CAD severity.

Effectiveness of board game activities for reducing depression among older adults in adult day care centers of Taiwan: a quasi-experimental study.

Depression is common in older adults and is associated with an increased risk of cognitive impairment. To clarify the possible roles of board game use in psychosomatic health promotion, this study evaluated the effects of board game activities in reducing depression in older adults. This was a quasi-experimental study. Purposive sampling was used to select 150 participants aged 65 years and above with intact mental functions who were currently residing in adult day care centers. Seventy-five participants who participated in 12 sessions of selected board game activities were assigned to the experimental group, and 75 participants who adhered to their ordinary activities were allocated to the control group. Structured questionnaires were used for data collection. The board game activities showed promising effects on the depression levels of the investigated older adults living in adult day care centers. Therefore, one possible beneficial effect of board game activities may be reduced depression in older adults. The results of this study provide support for the mediating role of board game activities in the mental health of long-term care elders. Incorporating board game activities into social work may help to make it more diverse and innovative.

Determination of the Pharmacokinetics and Tissue Distribution of Methyl 3,4-Dihydroxybenzoate (MDHB) in Mice Using Liquid Chromatography-Tandem Mass Spectrometry.

BACKGROUND AND OBJECTIVES: Methyl 3,4-dihydroxybenzoate (MDHB) has the potential to prevent neurodegenerative diseases (NDDs). The present work aims to reveal the pharmacokinetics and tissue distribution characteristics of MDHB. METHODS: The pharmacokinetics and tissue distribution of MDHB were analyzed using LC-MS/MS after a single intragastric administration (50 to 450 mg/kg) in mice, and samples were collected from five animals at specific time points. RESULTS: Pharmacokinetic parameters of MDHB following intragastric administrations were: the time to peak concentration (Tmax) ranged from 0.033 to 0.07 h, the peak concentration (Cmax) ranged from 12,379.158 to 109798.712 µg/l, the elimination half-life (t1/2z) ranged from 0.153 to 1.291 h, the area under the curve (AUC0-∞) ranged from 640.654 to 20,241.081 µg/l × h, the mean residence time (MRT0-∞) ranged from 0.071 to 0.206 h, the apparent volume of distribution (Vz/F) ranged from 17.538 to 45.244 l/kg, and the systemic clearance (Clz/F) ranged from 22.541 to 80.807 l/h/kg. The oral bioavailability of MDHB was 23%. The maximum MDHB content was detected in the stomach, and the minimum content was observed in the testes; the peak content in the brain was 15,666.93 ng/g. CONCLUSIONS: The pharmacokinetic characteristics of MDHB include fast absorption, high systemic clearance, a short half-life and an oral bioavailability of 23%. Additionally, MDHB permeates the blood-brain barrier (BBB) and is rapidly distributed to all organs. The identification of the pharmacokinetics of MDHB following its oral administration will contribute to further preclinical and clinical studies of its effects.

Effect of Yangzheng Sanjie Decoction on Proliferation, Apoptosis and ERK Pathway of Human Gastric Cancer MKN-45 Cells / 中国实验方剂学杂志

Objective: To investigate the effect of Yangzheng Sanjie decoction on proliferation, apoptosis and extracellular signal-regulated kinase (ERK) pathway of human gastric cancer MKN-45 cells.Method: Gastric cancer cell line MKN-45 was treated for 24, 48, 72 h with Yangzheng Sanjie decoction (0.5, 1, 1.5, 2, 2.5, 3, 3.5 g·L-1); cell proliferation was measured by cell counting kit-8 (CCK-8); cell colony forming ability was observed by the plate cloning experiment after intervention with Yangzheng Sanjie decoction (0.4, 0.8 g·L-1); MKN-45 cells was treated with 4, 8 g·L-1, and then cell apoptosis was detected by flow cytometry; the expression of ERK and its phosphorylation level were detected by Western blot assay after treatment with 2, 4, 8 g·L-1.Result: Compared with the blank group, Yangzheng Sanjie decoction could significantly inhibit the proliferation of MKN-45 cells. After treatment for 24, 48 h, Yangzheng Sanjie decoction started from 2 g·L-1, and after treatment for 72 h, it started from 1.5 g·L-1, the cell viability gradually decreased in a concentration-dependent manner (PPP-1, cell colonies could not be formed; the apoptosis rate of Yangzheng Sanjie decoction was significantly higher than that of the blank group (PP-1, and the phosphorylation level of ERK protein in MKN-45 cells was down-regulated (PConclusion: Yangzheng Sanjie decoction can inhibit the proliferation of human gastric cancer cell line MKN-45 and promote its apoptosis. The mechanism may be related to the inhibition of phosphorylation of ERK.

Novel subunit vaccine with linear array epitope protect giant grouper against nervous necrosis virus infection.

Viral nervous necrosis caused by nervous necrosis virus (NNV) is one of the most severe diseases resulting in high fish mortality rates and high economic losses in the giant grouper industry. Various NNV vaccines have been evaluated, such as inactivated viruses, virus-like particles (VLPs), recombinant coat proteins, synthetic peptides of coat proteins, and DNA vaccines. However, a cheaper manufacturing process and effective protection of NNV vaccines for commercial application are yet to be established. Hence, the present study developed a novel subunit vaccine composed of a carrier protein, receptor-binding domain of Pseudomonas exotoxin A, and tandem-repeated NNV coat protein epitopes by using the structural basis of epitope prediction and the linear array epitope (LAE) technique. On the basis of the crystal structure of the NNV coat protein, the epitope was predicted from the putative target cell receptor-binding region to elicit neutralizing immune responses. The safety of the LAE vaccine was evaluated, and all vaccinated fish survived without any physiological changes. The coat protein-specific antibody titers in the vaccinated fish increased after vaccine administration and exerted NNV-neutralizing effects. The efficacy tests revealed that the relative percent survival (RPS) of LAE antigen formulated with adjuvant was above 72% and LAE vaccine was effective for preventing NNV infection in giant grouper. This study is the first to develop an NNV vaccine by using epitope repeats, which provided effective protection to giant grouper against virus infection. The LAE construct can be used as a vaccine design platform against various pathogenic diseases.

Viral infection upregulates myostatin promoter activity in orange-spotted grouper (Epinephelus coioides).

Myostatin is a negative regulator of myogenesis and has been suggested to be an important factor in the development of muscle wasting during viral infection. The objective of this study was to characterize the main regulatory element of the grouper myostatin promoter and to study changes in promoter activity due to viral stimulation. In vitro and in vivo experiments indicated that the E-box E6 is a positive cis-and trans-regulation motif, and an essential binding site for MyoD. In contrast, the E-box E5 is a dominant negative cis-regulatory. The characteristics of grouper myostatin promoter are similar in regulation of muscle growth to that of other species, but mainly through specific regulatory elements. According to these results, we conducted a study to investigate the effect of viral infection on myostatin promoter activity and its regulation. The nervous necrosis virus (NNV) treatment significantly induced myostatin promoter activity. The present study is the first report describing that specific myostatin motifs regulate promoter activity and response to viral infection.

PI3K/Akt signaling pathway is involved in the neurotrophic effect of senegenin.

Neurodegenerative diseases are frequently associated with the loss of synapses and neurons. Senegenin, extracted from the Chinese herb Polygala tenuifolia Willd, was previously found to promote neurite outgrowth and neuronal survival in primary cultured rat cortical neurons. The aim of the present study was to investigate the underlying mechanisms of senegenin-induced neurotrophic effects on rat cortical neurons. Primary cortical rat neurons were treated with various pharmacological antagonists and with or without senegenin, and subjected to MTT and western blot analysis to explore the effects of senegenin on cell survival as well as the activation of signaling pathways. Neurite outgrowth and neuronal survival induced by senegenin were significantly inhibited by A2A receptor antagonist ZM241385 and specific phosphoinositide-3 kinase (PI3K) inhibitor LY294002, but not by tropomyosin receptor kinase A receptor inhibitor K252a, mitogen-activated protein kinase kinase inhibitor PD98059 or protein kinase C inhibitor GÖ6976. Furthermore, senegenin enhanced the phosphorylation of Akt, which was blocked by LY294002. The present study revealed that the PI3K/Akt signaling pathway may be involved in the neurotrophic effects of senegenin.

Methyl 3,4-dihydroxybenzoate extends the lifespan of Caenorhabditis elegans, partly via W06A7.4 gene.

To identify and analyze the compounds that delay aging and extend the lifespan is an important aspect of the gerontology research. A number of compounds, including the ones with the antioxidant properties, have been shown to extend the lifespan of Caenorhabditis elegans. Here, we report that methyl 3,4-dihydroxybenzoate (MDHB), a small antioxidant molecule, prolongs the C. elegans' lifespan under normal as well as stress conditions, delays the age-associated decline in the pharyngeal pumping rate, and obviously enhances the abilities of scavenging intracellular reactive oxygen species (ROS). To further investigate the mechanism underlying the anti-aging action of MDHB, microarray analyses were performed, which demonstrated that 13 genes were differentially expressed in worms treated with MDHB for 48 and 144 h in common. RNA interference of W06A7.4 (NM_001269697.1), the most significantly up-regulated gene, shortened the lifespan of worms by 14%, compared with the L4440 control. Our findings demonstrate that W06A7.4 is a potentially positive determinant of the MDHB induced C. elegans' lifespan extension effect.

Neuroprotective effects of methyl 3,4-dihydroxybenzoate against H2O2-induced apoptosis in RGC-5 cells.

In the present study, we investigated the protective effect of methyl 3,4-dihydroxybenzoate (MDHB) against H2O2-induced apoptosis in RGC-5 cells. The RGC-5 cells were cultured in plates for 24 h, which were then pretreated with dimethyl sulfoxide, different concentrations of MDHB, or probucol for 12 h prior to addition of 300 µM H2O2 for 24 h. The cell viability was detected by MTT assay. The rate of apoptosis, level of lipid peroxidation, and mitochondrial membrane potential (MMP) were detected by flow cytometry. Western blot analysis was also used to measure the expression level of Bcl-2, Bax, caspase 9, and caspase 3 proteins in H2O2-treated RGC-5 cells. Our study showed that the cell viability of RGC-5 cells significantly decreased after treatment with 300 µM H2O2 for 24 h, but MDHB (8, 16, 32 µM) increased RGC-5 cell survival, suppressed the rate of apoptosis, scavenged reactive oxygen species, and restored MMP. MDHB also obstructed H2O2-induced apoptosis by regulating the expression of Bcl-2 and Bax, as well as suppressing the activation of caspase 9 and caspase 3. Our results showed that MDHB is an effective neuroprotective compound that mitigates oxidative stress and inhibits apoptosis in RGC-5 cells.

Methyl 3,4-dihydroxybenzoate protects primary cortical neurons against Aß25₋35-induced neurotoxicity through mitochondria pathway.

Amyloid-ß peptides (Aß), which can aggregate into oligomers or fibrils in neurons, play a critical role in the pathogenesis of Alzheimer's disease (AD). Methyl 3,4-dihydroxybenzoate (MDHB), a phenolic acid compound, has been reported to have antioxidative and neurotrophic effects. The present study investigated the neuroprotective effects of MDHB against Aß-induced apoptosis in rat primary cortical neutons. The primary cortical neurons were pretreated with different concentrations of MDHB for 24 hr, then incubated with 10 µM Aß25-35 for 24 hr. The results showed that Aß25-35 could induce neurotoxicity as evidenced by the decreased cell viability and the increased apoptotic rate. In parallel, Aß25-35 significantly increased the reactive oxygen species accumulation and decreased mitochondrial membrane potential. However, pretreatment of the primary cortical neurons with MDHB could effectively suppress these cellular events caused by Aß25-35 exposure. In addition, MDHB could increase the level of Bcl-2, decrease the level of Bax, and inhibit the activation of caspase-9 and caspase-3 in Aß25-35 -treated primary cortical neurons. All these results were beneficial in their protective effect against Aß-induced neurotoxicity. Our results suggest that MDHB has a neuroprotective effect that provides a pharmacological basis for its clinical use in the treatment of AD.

Stable integration and expression of heterologous genes in several lactobacilli using an integration vector constructed from the integrase and attP sequences of phage ΦAT3 isolated from Lactobacillus casei ATCC 393.

An integration vector capable of stably integrating and maintaining in the chromosomes of several lactobacilli over hundreds of generations has been constructed. The major integration machinery used is based on the ΦAT3 integrase (int) and attP sequences determined previously. A novel core sequence located at the 3' end of the tRNA(leu) gene is identified in Lactobacillus fermentum ATCC 14931 as the integration target by the integration vector though most of such sequences found in other lactobacilli are similar to that determined previously. Due to the lack of an appropriate attB site in Lactococcus lactis MG1363, the integration vector is found to be unable to integrate into the chromosome of the strain. However, such integration can be successfully restored by cotransforming the integration vector with a replicative one harboring both attB and erythromycin resistance sequences into the strain. Furthermore, the integration vector constructed carries a promoter region of placT from the chromosome of Lactobacillus rhamnosus TCELL-1 which is used to express green fluorescence and luminance protein genes in the lactobacilli studied.

[Health issues and preventive health strategies for adolescent girls].

As adolescent girls have specific healthcare needs, this paper was designed to provide a better understanding of their healthcare needs in both physical and psychosocial terms. After conducting a targeted review of the literature on children and adolescents, we identified factors of importance to physical health as body weight, physical activity, menstruation, sexual knowledge and attitude and to psychosocial health as anxiety, interpersonal relationships, depression, and suicide behavior. Reflecting these factors, this paper presents four preventive suggestions to clinical practice, education, and research to facilitate improvements in adolescent girl's health. These suggestions include: 1) strengthening health education and media responsibility with regard to adolescent girl health; 2) improving awareness of the needs of adolescent girls within healthcare and education organizations; 3) making health guidelines for promoting proper health behavior in adolescent girls; and 4) mobilizing nurses to assert the health of adolescent girls in clinical, education, and research fields. This is the first paper that focuses on the health needs of adolescent Taiwanese girls. The authors hope that more people become involved in the care of adolescent girl's health in Taiwan.

Estimating low-toxic-effect concentrations in closed-system algal toxicity tests.

The no-observed-effect concentrations (NOEC) and EC(10) values for 108 organic compounds were estimated, using multiple endpoints (i.e., biopopulation, growth rate, and dissolved oxygen production), from previous data obtained by a closed-system algal toxicity test (test alga: Pseudokirchneriella subcapitata). These low-toxic-effect concentrations are valuable to risk assessment of chemicals and protection of the aquatic environment as such information is quite scarce in existing toxicological databases. Furthermore, based on limited amount of available data, we found that the risk of organic toxicants to phytoplankton may be severely underestimated by existing databases, which are primarily derived by the conventional batch technique. Good correlation relationships between NOEC (or EC(10)) and EC(50) values were established. For polar and nonpolar narcotics, quantitative structure-activity relationships (QSARs) based on hydrophobicity, and/or the lowest unoccupied molecular orbital energy (Elumo) were developed with satisfactory predictive powers. The above statistical relationships can be applied to derive a preliminary estimation for the low-toxic-effect levels for other (or new) organic compounds that has no toxicological data available.

Formation of an inverted repeat junction in the transposition of insertion sequence ISLC3 isolated from Lactobacillus casei.

An insertion sequence, ISLC3, of 1351 bp has been isolated from Lactobacillus casei. Formation of IS circles containing a 3 bp spacer (complete junction) or deletion of 25 bp at the left inverted repeat (IRL) between the abutted IS ends of the ISLC3 junction region (deleted junction) was also discovered in the lactobacilli and Escherichia coli system studied. We found that the promoter formed by the complete junction P(jun) was more active than that formed by the 25 bp deleted junction P(djun) or the indigenous promoter P(IRL). The corresponding transcription start sites for both promoter P(jun) and P(IRL) as well as P(djun) were subsequently determined using a primer extension assay. The activity of transposase OrfAB of ISLC3 was also assayed using an in vitro system. It was found that this transposase preferred to cleave a single DNA strand at the IRR over the IRL end in the transposition process, suggesting that attack of one end by the other was oriented from IRR to IRL.

Complete genomic sequence of the temperate bacteriophage PhiAT3 isolated from Lactobacillus casei ATCC 393.

The complete genomic sequence of a temperate bacteriophage PhiAT3 isolated from Lactobacillus (Lb.) casei ATCC 393 is reported. The phage consists of a linear DNA genome of 39,166 bp, an isometric head of 53 nm in diameter, and a flexible, noncontractile tail of approximately 200 nm in length. The number of potential open reading frames on the phage genome is 53. There are 15 unpaired nucleotides at both 5' ends of the PhiAT3 genome, indicating that the phage uses a cos-site for DNA packaging. The PhiAT3 genome was grouped into five distinct functional clusters: DNA packaging, morphogenesis, lysis, lysogenic/lytic switch, and replication. The amino acid sequences at the NH2-termini of some major proteins were determined. An in vivo integration assay for the PhiAT3 integrase (Int) protein in several lactobacilli was conducted by constructing an integration vector including PhiAT3 int and the attP (int-attP) region. It was found that PhiAT3 integrated at the tRNAArg gene locus of Lactobacillus rhamnosus HN 001, similar to that observed in its native host, Lb. casei ATCC 393.

Evaluation of murine models of permanent focal cerebral ischemia.

BACKGROUND: To date murine models of permanent focal cerebral ischemia have not been well characterized. The purposes of this paper were to compare three different permanent middle cerebral artery occlusion (MCAo) models with or without craniectomy, and to identify an ideal mouse model of permanent focal cerebral ischemia. METHODS: Experiments were performed on 45 healthy adult male Kunming mice, weighing 28 to 42 g. The animals were randomly assigned to three groups (n = 15 in every group) based on surgical procedure: MCAo via the external carotid artery (ECA), MCAo via the common carotid artery (CCA), and direct ligation of the middle cerebral artery (MCA). Each day post-ischemia, the animals were scored using an eight-grade neurological function scale, and mortality was also recorded. Seven days post-ischemia, the brains were removed for lesion size determination using triphenyltetrazolium chloride staining. Correlation analysis of lesion volume and neurological score was carried out. RESULTS: Mortality in the group receiving direct MCA ligation was lowest among the three groups, and there was a significant difference between the direct MCA ligation group and the two intraluminal occlusion groups (P < 0.05). In all groups, neurological scores gradually increased with prolongation of ischemic duration, peaking after two days, then gradually decreasing. In the direct MCA ligation group, however, neurological scores were relatively stable. There was a significant correlation between infarct volume and neurological score 7 days after MCAo in every group (all r > 0.7, P < 0.05), suggesting good reproducibility of lesion volume in the three groups, but the infarct volume was more constant in the direct MCA ligation group. CONCLUSION: The direct ligation model of MCAo provides an optimal means of studying permanent focal cerebral ischemia, and is preferable to the models using intraluminal sutures.

Evaluation of murine models of permanent focal cerebral ischemia / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>To date murine models of permanent focal cerebral ischemia have not been well characterized. The purposes of this paper were to compare three different permanent middle cerebral artery occlusion (MCAo) models with or without craniectomy, and to identify an ideal mouse model of permanent focal cerebral ischemia.</p><p><b>METHODS</b>Experiments were performed on 45 healthy adult male Kunming mice, weighing 28 to 42 g. The animals were randomly assigned to three groups (n = 15 in every group) based on surgical procedure: MCAo via the external carotid artery (ECA), MCAo via the common carotid artery (CCA), and direct ligation of the middle cerebral artery (MCA). Each day post-ischemia, the animals were scored using an eight-grade neurological function scale, and mortality was also recorded. Seven days post-ischemia, the brains were removed for lesion size determination using triphenyltetrazolium chloride staining. Correlation analysis of lesion volume and neurological score was carried out.</p><p><b>RESULTS</b>Mortality in the group receiving direct MCA ligation was lowest among the three groups, and there was a significant difference between the direct MCA ligation group and the two intraluminal occlusion groups (P < 0.05). In all groups, neurological scores gradually increased with prolongation of ischemic duration, peaking after two days, then gradually decreasing. In the direct MCA ligation group, however, neurological scores were relatively stable. There was a significant correlation between infarct volume and neurological score 7 days after MCAo in every group (all r > 0.7, P < 0.05), suggesting good reproducibility of lesion volume in the three groups, but the infarct volume was more constant in the direct MCA ligation group.</p><p><b>CONCLUSION</b>The direct ligation model of MCAo provides an optimal means of studying permanent focal cerebral ischemia, and is preferable to the models using intraluminal sutures.</p>