[Safety and efficacy of the early administration of levosimendan in patients with acute non-ST-segment elevation myocardial infarction and elevated NT-proBNP levels: An Early Management Strategy of Acute Heart Failure (EMS-AHF)].

Objectives: To investigated the safety and efficacy of treating patients with acute non-ST-segment elevation myocardial infarction (NSTEMI) and elevated levels of N-terminal pro-hormone B-type natriuretic peptide (NT-proBNP) with levosimendan within 24 hours of first medical contact (FMC). Methods: This multicenter, open-label, block-randomized controlled trial (NCT03189901) investigated the safety and efficacy of levosimendan as an early management strategy of acute heart failure (EMS-AHF) for patients with NSTEMI and high NT-proBNP levels. This study included 255 patients with NSTEMI and elevated NT-proBNP levels, including 142 males and 113 females with a median age of 65 (58-70) years, and were admitted in the emergency or outpatient departments at 14 medical centers in China between October 2017 and October 2021. The patients were randomly divided into a levosimendan group (n=129) and a control group (n=126). The primary outcome measure was NT-proBNP levels on day 3 of treatment and changes in the NT-proBNP levels from baseline on day 5 after randomization. The secondary outcome measures included the proportion of patients with more than 30% reduction in NT-proBNP levels from baseline, major adverse cardiovascular events (MACE) during hospitalization and at 6 months after hospitalization, safety during the treatment, and health economics indices. The measurement data parameters between groups were compared using the t-test or the non-parametric test. The count data parameters were compared between groups using the χ² test. Results: On day 3, the NT-proBNP levels in the levosimendan group were lower than the control group but were statistically insignificant [866 (455, 1 960) vs. 1 118 (459, 2 417) ng/L, Z=-1.25,P=0.21]. However, on day 5, changes in the NT-proBNP levels from baseline in the levosimendan group were significantly higher than the control group [67.6% (33.8%,82.5%)vs.54.8% (7.3%,77.9%), Z=-2.14, P=0.03]. There were no significant differences in the proportion of patients with more than 30% reduction in the NT-proBNP levels on day 5 between the levosimendan and the control groups [77.5% (100/129) vs. 69.0% (87/126), χ²=2.34, P=0.13]. Furthermore, incidences of MACE did not show any significant differences between the two groups during hospitalization [4.7% (6/129) vs. 7.1% (9/126), χ²=0.72, P=0.40] and at 6 months [14.7% (19/129) vs. 12.7% (16/126), χ²=0.22, P=0.64]. Four cardiac deaths were reported in the control group during hospitalization [0 (0/129) vs. 3.2% (4/126), P=0.06]. However, 6-month survival rates were comparable between the two groups (log-rank test, P=0.18). Moreover, adverse events or serious adverse events such as shock, ventricular fibrillation, and ventricular tachycardia were not reported in both the groups during levosimendan treatment (days 0-1). The total cost of hospitalization [34 591.00(15 527.46,59 324.80) vs. 37 144.65(16 066.90,63 919.00)yuan, Z=-0.26, P=0.80] and the total length of hospitalization [9 (8, 12) vs. 10 (7, 13) days, Z=0.72, P=0.72] were lower for patients in the levosimendan group compared to those in the control group, but did not show statistically significant differences. Conclusions: Early administration of levosimendan reduced NT-proBNP levels in NSTEMI patients with elevated NT-proBNP and did not increase the total cost and length of hospitalization, but did not significantly improve MACE during hospitalization or at 6 months.

[Genetic analysis and prenatal diagnosis of Duchenne or Becker muscular dystrophy].

Objective: To explore the mutation characteristics of DMD gene in patients with Duchenne or Becker muscular dystrophy and female carriers, to provide effective prenatal diagnosis. Methods: Samples were collected from 94 male patients clinically diagnosed with Duchenne or Becker muscular dystrophy and 121 corresponding female relatives from Qingdao Women and Children's Hospital from June 2011 to October 2018. Multiplex ligation-dependent probe amplification (MLPA) was used to detect their DMD gene, and 23 high risk pregnants were performed prenatal diagnosis. Any candidate of DMD gene single-exon deletion was validated by further PCR amplification. The sample with whole DMD gene deletion was confirmed by chromosomal microarray analysis (CMA) to detect copy number variations and break site. Results: Among 94 clinical Duchenne or Becker muscular dystrophy patients, 66(70.2%, 66/94) were detected gene mutation; 56 cases were exon deletion mutation and 10 cases were duplication mutation. In 121 female relatives, 48 cases (39.7%, 48/121) were diagnosed as carriers. The mutation carrying rate, was 64.5% (40/62) identified in 62 mothers of Duchenne or Becker muscular dystrophy patients. Five Duchenne or Becker muscular dystrophy fetuses and 5 carrier fetuses were prenatally diagnosed in 23 high risk pregnants. Two children with the entire DMD gene deletion were identified more deletions at Xp21, with deletions of 6.66 Mb and 10.64 Mb respectively. Conclusions: MLPA may be an important method to detect DMD gene mutation of deletion and duplication. Therefore, the diagnosis of probands, female carriers and making an effective prenatal diagnosis are essential to reduce the birth of children with Duchenne or Becker muscular dystrophy.

The relationship between clinical characteristics including presence of exposed lesions and health-related quality of life (HRQoL) in patients with psoriasis: analysis from the nationwide epidemiologic study for psoriasis in Korea (EPI-PSODE study).

BACKGROUND: Psychological aspect and quality of life should be considered in treating patients with psoriasis. OBJECTIVE: We sought to ascertain which clinical characteristics including presence of exposed lesions are associated with impairment of health-related quality of life (HRQoL) in patients with psoriasis. METHODS: The EPI-PSODE study was a nationwide, multicenter, cross-sectional study conducted in Korea that included 1260 adult patients with psoriasis. In addition to clinical characteristics including presence of exposed lesions, data were collected using the Psoriatic Arthritis (PsA) Screening and Evaluation (PASE), Dermatology Life Quality Index (DLQI), MOS 36-Item Short-Form Health Survey (SF-36), Work Productivity and Activity Impairment Questionnaire Psoriasis (WPAI: PSO) and Medication Satisfaction Questionnaire (MSQ). RESULTS: Patients with a DLQI score > 5 (n = 990) were younger, had an earlier onset of psoriasis, scored higher on the Psoriasis Area and Severity Index (PASI), had higher body surface area (BSA) and had higher PASE scores than patients with DLQI ≤ 5 (n = 266). The group of patients with exposed lesions (n = 871) were younger and male predominance, earlier onset of psoriasis, longer disease duration, higher PASI/BSA score and a higher proportion with drinking and smoking history each than the group of patients without exposed lesions (n = 389). Presence of exposed lesions negatively influenced DLQI, 36-Item Short-Form Health Survey (SF-36) (mental component), presenteeism, total work productivity impairment and total activity impairment in the WPAI: PSO. In multiple regression model, PASI score was the only variable which was significantly associated with all HRQoL measures. Presence of exposed lesions was a significant factor affecting DLQI and SF-36 (mental). CONCLUSION: The presence of exposed lesions has a negative impact on quality of life, mental health and work productivity. Therefore, effective treatments are particularly needed for psoriasis patients with exposed lesions.

Maternal dietary linoleic acid supplementation promotes muscle fibre type transformation in suckling piglets.

As meat quality is basically dependent on muscle fibre characteristics, it is important to know how muscle fibres are regulated and transformed. This study aimed to investigate the effect of maternal dietary supplementation on muscle fibre types using 3% saturated fatty acid (palmitic acid, PA) or 3% unsaturated fatty acid (linoleic acid, LA) from 80 days of gestation to the weaning of offspring (25 days post-natal). The results indicated that higher mRNA levels of MyHCI type genes were found in the soleus muscles of piglets that suckled from LA-supplemented sows than from PA-supplemented sows. In addition, LA treatment increased the gene expression of the type I muscle fibre marker troponin I (p < 0.01), suggesting that LA promoted muscle fibre type transformation to type I fibres. Moreover, PGC-1α (p < 0.01) and MEF2c (p < 0.05) mRNA levels were higher in the piglets from the LA treatment group than in those from the PA treatment group. Furthermore, LA supplementation also significantly increased AMP-activated protein kinase (AMPK) mRNA levels (p < 0.05), which is an upstream regulator of PGC-1α. Collectively, these findings demonstrated that maternal dietary LA supplementation promoted muscle fibre transformation to type I fibre and that this process may be mediated through an AMPK-dependent pathway.

Effect of ultrasonic waves on the freezing rates of potatoes in degassed coolant and untreated coolant.

BACKGROUND: Ultrasonic waves are shown to enhance the rate of freezing. OBJECTIVE: To elucidate the mechanism of immersion freezing of potatos with ultrasonic waves. MATERIALS AND METHODS: Ultrasound is applied to potato samples immersed in degassed coolant and untreated coolant. Sonic waves were intermittently applied at temperature below -1 degree C. The freezing rates were measured under different experimental conditions. RESULTS: The use of ultrasonic waves increased the freezing rates of potatoes immersed in both degassed coolant and untreated coolant. However, the freezing rate in the degassed coolant was less than that in the untreated coolant. CONCLUSION: Heat transfer on the interface between the potato sample and sonicated degassed coolant appears to be less than that within the sample in the absence of cavitation. The interface heat transfer between the potato sample and untreated coolant is likely improved due to ultrasonic cavitation.

Influence of dietary inclusion of Bacillus licheniformis on laying performance, egg quality, antioxidant enzyme activities, and intestinal barrier function of laying hens.

This experiment was conducted to evaluate the effects of dietary inclusion of Bacillus licheniformis on laying performance, egg quality, antioxidant enzyme activities, and intestinal barrier function of laying hens. Hy-Line Variety W-36 hens (n = 540; 28 wk of age) were randomized into 6 groups, each group with 6 replications (n = 15). The control group received the basal diet formulated with maize and soybean meal. The treatment groups received the same basal diets supplemented with 0.01, 0.02, 0.03, 0.06, and 0.09% Bacillus licheniformis powder (2 × 10(10) cfu/g) for an 8-wk trial. The results showed that dietary supplementation with 0.01 and 0.03% B. licheniformis significantly increased egg production and egg mass. However, no significant differences were observed in egg weight, feed consumption, and feed conversion efficiency among the 6 groups. Supplementation with different levels of B. licheniformis was found to be effective in improvement of egg quality by increasing egg shell thickness and strength. Compared with control, d-lactate content, diamine oxidase activity, and adrenocorticotropic hormone level in serum decreased significantly, and the level of estradiol and follicle-stimulating hormone increased significantly in plasma of all the experimental groups. Dietary supplementation with B. licheniformis increased the intestinal villus height and reduced the crypt depth. In conclusion, dietary inclusion of B. licheniformis could improve laying performance and egg quality significantly in a dose-dependent manner by decreasing the stress response, upregulating the growth hormone, and improving intestinal health.

Effect of Saccharomyces boulardii and Bacillus subtilis B10 on intestinal ultrastructure modulation and mucosal immunity development mechanism in broiler chickens.

The recent ban on the use of antibiotics as a feed additive has led to the search for alternative sources of antibiotics in the feed industry. Presently, probiotics are considered as a potential substitute for antibiotic as a live biotherapeutic agent to improve animal health and performance. Accordingly, study was focused on evaluating the effect of Saccharomyces boulardii (Sb) and Bacillus subtilis B10 (Bs) on ultrastructure modulation and mucosal immunity development in broiler chickens. A total of three hundred 1-d-old Sanhuang broilers (a Chinese cross breed) were randomized into 3 groups, each group with 5 replications (n = 20). The control group (Ctr) was fed a basal diet containing an antibiotic (virginiamycin, 20 mg/kg). Meanwhile, broilers in experimental groups received Sb and Bs (1 × 10(8) cfu/kg of feed) in addition to the basal diet for 72 d. The results of the experimental groups revealed a significant improvement in live BW and relative weight of bursa of Fabricius and thymus. Also, intestinal villus height, width, and number of goblet cells increased in the Sb and Bs groups. Meanwhile, modulation in the intestinal ultrastructure and increased mRNA expression levels of occluding, cloudin2, and cloudin3 (P < 0.05) were observed in the Sb and Bs groups. Moreover, IgA-positive cells significantly increased in the jejunum of Sb- and Bs-supplemented groups (P < 0.05). Intestinal cytokines interleukin-6, tumor necrosis factor-α, interleukin-10, transforming growth factor-ß, and secretory IgA concentrations were (P < 0.05) improved in the probiotic groups; however, Sb induced inflammatory and antiinflammatory cytokines (P < 0.05) in comparison with the Ctr group. The present findings conclusively revealed that Sb and Bs increased IgA-positive cells in the lumen of the intestinal villus and revealed that Sb and Bs could modulate intestinal ultrastructure through increasing occluding, cloudin2, and cloudin3 mRNA expression levels in broiler intestine.

Correlation between retinal vein pulse amplitude, estimated intracranial pressure, and postural change.

Spaceflight associated neuro-ocular syndrome (SANS) is common amongst astronauts on long duration space missions and is associated with signs consistent with elevated cerebrospinal fluid (CSF) pressure. Additionally, CSF pressure has been found to be elevated in a significant proportion of astronauts in whom lumbar puncture was performed after successful mission completion. We have developed a retinal photoplethysmographic technique to measure retinal vein pulsation amplitudes. This technique has enabled the development of a non-invasive CSF pressure measurement apparatus. We tested the system on healthy volunteers in the sitting and supine posture to mimic the range of tilt table extremes and estimated the induced CSF pressure change using measurements from the CSF hydrostatic indifferent point. We found a significant relationship between pulsation amplitude change and estimated CSF pressure change (p < 0.0001) across a range from 2.7 to 7.1 mmHg. The increase in pulse amplitude was highest in the sitting posture with greater estimated CSF pressure increase (p < 0.0001), in keeping with physiologically predicted CSF pressure response. This technique may be useful for non-invasive measurement of CSF pressure fluctuations during long-term space voyages.

Evaluating the impact of excess dietary tryptophan on laying performance and immune function of laying hens reared under hot and humid summer conditions.

1. Tryptophan (Trp), besides its role as an essential amino acid in protein synthesis, may also have other important effects on laying hens under summer conditions. 2. Babcock Brown layers (n = 768), 40 weeks of age, were allocated to 4 treatment groups, each of which included 6 replicates of 32 hens. Each group received the same basal diet, formulated with maize and soybean meal, for 8 weeks. Hens were fed on the basal diet with 0·0, 0·2, 0·4, and 0·8 g/kg L-Trp to achieve dietary concentrations of 1·7, 1·9 g/kg, 2·1 g/kg or 2·5 g/kg of Trp, respectively. 3. Supplementing L-Trp had no affect on laying performance. Adding 0·2 or 0·4 g/kg L-Trp improved egg shell strength compared with those fed on the control diet. Serum albumin concentration increased at 0·4 g/kg compared with those receiving 0·0 or 0·8 g/kg Trp. The addition of Trp at 0·4 g/kg increased serum IgM concentration quadratically. Serum superoxide dismutase (T-SOD) increased linearly and quadratically at 0·4 g/kg. 4. In conclusion, we suggest that 0·2 to 0·4 g/kg Trp may have beneficial effects on laying hens under conditions of high temperature and humidity.

Zero retinal vein pulsation amplitude extrapolated model in non-invasive intracranial pressure estimation.

Intracranial pressure (ICP) includes the brain, optic nerve, and spinal cord pressures; it influences blood flow to those structures. Pathological elevation in ICP results in structural damage through various mechanisms, which adversely affects outcomes in traumatic brain injury and stroke. Currently, invasive procedures which tap directly into the cerebrospinal fluid are required to measure this pressure. Recent fluidic engineering modelling analogous to the ocular vascular flow suggests that retinal venous pulse amplitudes are predictably influenced by downstream pressures, suggesting that ICP could be estimated by analysing this pulse signal. We used this modelling theory and our photoplethysmographic (PPG) retinal venous pulse amplitude measurement system to measure amplitudes in 30 subjects undergoing invasive ICP measurements by lumbar puncture (LP) or external ventricular drain (EVD). We estimated ICP from these amplitudes using this modelling and found it to be accurate with a mean absolute error of 3.0 mmHg and a slope of 1.00 (r = 0.91). Ninety-four percent of differences between the PPG and invasive method were between - 5.5 and + 4.0 mmHg, which compares favourably to comparisons between LP and EVD. This type of modelling may be useful for understanding retinal vessel pulsatile fluid dynamics and may provide a method for non-invasive ICP measurement.

Organic trace minerals improve eggshell quality by improving the eggshell ultrastructure of laying hens during the late laying period.

The objective of this study was to investigate the effects of low inclusion levels of organic trace minerals (iron, copper, manganese, and zinc) on performance, eggshell quality, serum hormone levels, and enzyme activities of laying hens during the late laying period. A total of 405 healthy hens (HY-Line White, 50-week-old) were randomly divided into 3 treatments, with 9 replications per treatment and 15 birds per replication. The dietary treatments included a basal diet supplemented with inorganic trace minerals at commercial levels (CON), a basal diet supplemented with inorganic trace minerals at 1/3 commercial levels (ITM), and a basal diet supplemented with proteinated trace minerals at 1/3 commercial levels (TRT). The trial lasted 56 D (8 wk). Compared with the CON group, the ITM group showed decrease in (P < 0.05) egg production, eggshell strength, eggshell palisade layer, palisade layer ratio, serum estrogen, luteinizing hormone, glycosaminoglycan concentration, and carbonic anhydrase activity and increase in (P < 0.05) egg loss and mammillary layer ratio. However, the TRT group almost kept all the indices close to the CON group (P > 0.05). Furthermore, hens fed with low inclusion levels of organic trace minerals had smaller mammillary knobs (P < 0.05) than those in the CON and ITM groups. In conclusion, hens fed with low inclusion levels of proteinated trace minerals had better performance and eggshell strength than those fed with identical levels of inorganic compounds; organic trace minerals improved eggshell quality by improving the eggshell ultrastructure of laying hens during the late laying period.

HCP5 promotes colon cancer development by activating AP1G1 via PI3K/AKT pathway.

OBJECTIVE: To explore whether HCP5 participates in the pathogenic progression of colon cancer (CC) and its underlying mechanism. PATIENTS AND METHODS: HCP5 expression in CC tissues and cell lines was detected by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR). The correlation between the HCP5 expression and tumor stage of CC patients was then analyzed. After CC cells were transfected with HCP5-siRNA, the proliferation and migration capacities were detected by cell counting kit-8 (CCK-8), colony formation and transwell assay, respectively. Cell cycle was examined by flow cytometry. Western blot was conducted to detect protein expressions of HCP5, AP1G1 and relative molecules in the PI3K/AKT pathway. Rescue experiments were performed by co-transfection of HCP5-siRNA and AP1G1-siRNA into CC cells, followed by cell function detection. RESULTS: HCP5 was highly expressed, whereas AP1G1 was lowly expressed in CC tissues and cell lines. Besides, CC patients with stage III-IV presented higher expression of HCP5 than those with stage I-II. The knockdown of HCP5 in CC cells down-regulated proliferation and migration capacities, and arrested cell cycle in the G0/G1 phase, which was reversed by the AP1G1 knockdown. In addition, HCP5 knockdown up-regulated AP1G1 expression, whereas down-regulated the expression of relative proteins in the PI3K/AKT pathway. CONCLUSIONS: HCP5 was significantly increased in CC and enhanced the proliferation and migration of CC cells by inhibiting the AP1G1 expression. HCP5 promoted CC development by activating the PI3K/AKT pathway.

Safety evaluation of zinc methionine in laying hens: Effects on laying performance, clinical blood parameters, organ development, and histopathology.

The study was conducted to investigate whether high-dose zinc methionine (Zn-Met) affected the safety of laying hens, including laying performance, hematological parameters, serum chemical parameters, organ index, and histopathology. A total of 540 20-week-old Hy-Line White laying hens was randomly allocated to 6 groups with 6 replicates of 15 birds each. Birds were fed diets supplemented with 0 (control), 70, 140, 350, 700, or 1,400 mg Zn/kg diet as Zn-Met. The experiment lasted for 8 wk after a 2-week acclimation period. Results showed that dietary supplementation with 70 or 140 mg Zn/kg diet as Zn-Met significantly increased average daily egg mass (ADEM), laying rate (LR), and feed conversion ratio (FCR) (P < 0.05) and lowered broken and soft-shelled egg ratio (BSER) (P < 0.05) in comparison with the control group; no significant differences were detected among hens fed with 0, 350, or 700 mg Zn/kg as Zn-Met (P > 0.05); hens administered 1,400 mg Zn/kg showed a significant increase in BSER and remarkable decreases in ADEM, LR, and FCR (P < 0.001). There were no significant differences among hens receiving 0, 70, 140, 350, or 700 mg Zn/kg as Zn-Met in serum chemical parameters (P > 0.05); supplementation with 1,400 mg Zn/kg as Zn-Met remarkably elevated the concentrations of serum total bilirubin (TBILI), glucose (GLU), uric acid (UA), and creatinine (CRE) (P < 0.001), and enhanced activities of serum glutamic oxalacetic transaminase (GOP) and alkaline phosphatase (AKP) (P < 0.001) compared with the control group. No significant histopathological changes were found in hens administered 0, 70, 140, 350, or 700 mg Zn/kg as Zn-Met, while significant histological lesions were observed in the heart, liver, lung, and kidney tissues of hens receiving 1,400 mg Zn/kg as Zn-Met. No significant differences were detected in hematological parameters or organ index (P > 0.05). In conclusion, a nominal Zn concentration of 700 mg/kg as Zn-Met is considered to be no-observed-adverse-effect level following daily administration to hens for 56 days.

Antibiotic inclusion in the diet did not alter the standardized ileal digestible tryptophan to lysine ratio for growing pigs.

Two 21-d experiments were conducted to determine the optimum standardized ileal digestible (SID) Trp:Lys ratio for growing pigs; 1 experiment fed diets supplemented with or without an antibiotic. The primary response variables in both experiments were ADG, ADFI, G:F, and plasma urea N (PUN) concentrations with the optimum SID Trp:Lys ratio detected using broken-line analysis. Experiment 1 evaluated the optimum SID Trp:Lys ratio in growing pigs fed diets supplemented with an antibiotic. This experiment used 120 crossbred pigs that were blocked by sex and initial BW (24.13 ± 2.72 kg) and allotted to 6 SID Trp:Lys ratios in 4 replicates. Dietary treatments were formulated by the addition of crystalline Trp to create 6 SID Trp:Lys ratios (13.08%, 14.06%, 15.04%, 17.00%, 18.95%, and 20.91%) with a constant SID Lys level of 0.655%. As SID Trp:Lys ratios increased, ADG, ADFI, and G:F increased, and PUN concentrations decreased linearly ( < 0.05) and quadratically ( < 0.05). Linear broken-line analysis yielded optimum SID Trp:Lys ratios of 17.93% ( < 0.001) and 16.17% ( = 0.009) for ADG and PUN, respectively, resulting in a mean optimum SID Trp:Lys ratio of 17.05%. Experiment 2 evaluated the optimum SID Trp:Lys ratio in growing pigs fed diets supplemented with or without an antibiotic. It used a total of 324 crossbred pigs (initial BW: 30.81 ± 3.56 kg) that were allotted to 6 SID Trp:Lys ratios in 6 replicates. Dietary treatments were formulated by the addition of crystalline Trp to create 6 SID Trp:Lys ratios (12.52%, 14.86%, 17.20%, 19.54%, 21.88%, and 24.22%) with a constant SID Lys level of 0.67%. As SID Trp:Lys ratios increased, ADG, ADFI, and G:F increased, and PUN concentrations decreased linearly ( < 0.001) and quadratically ( < 0.001) regardless of antibiotic inclusion. There were no differences by the antibiotic treatment in ADG, ADFI, G:F, or PUN concentrations ( > 0.49) and no interactions between antibiotics and Trp:Lys ratios ( > 0.29). When the data for all pigs were pooled for the various Trp:Lys ratios, the optimum SID Trp:Lys ratios for ADG and PUN based on linear broken-line analysis were 14.58% ( < 0.001) and 14.54% ( < 0.001), respectively, resulting in an optimum SID Trp:Lys ratio of 14.56% as the mean of the determined optima for ADG and PUN responses. These results demonstrate that the optimum SID Trp:Lys ratio for 30- to 50-kg growing pigs is not impacted by the dietary inclusion of an antibiotic as long as the diets are formulated on an SID AA basis.

Determination of a threshold compromise value for the perfusion index by laser Doppler imaging after digital revascularization.

We have used laser Doppler imaging to monitor the microcirculation of replanted digits during the post-operative period in 103 patients who underwent either replantation after traumatic amputation or toe-to-finger reconstruction. The blood flow (perfusion unit) in each revascularized digit was compared with that of an unaffected digit. The perfusion index was defined as the perfusion value of a revascularized digit divided by the perfusion value of the neighbouring normal digit. The ideal threshold value of the perfusion index (0.397) was calculated by determining the receiver operating characteristic curve with optimal sensitivity and specificity. The corresponding Youden's index was 0.828. We believe that by establishing a threshold, that laser Doppler imaging should provide a reliable and objective assessment for the development of perfusion compromise in revascularized digits. LEVEL OF EVIDENCE: III.

Overexpression of membrane-type matrix metalloproteinase-1 gene induces mammary gland abnormalities and adenocarcinoma in transgenic mice.

To investigate the role of membrane-type matrix metalloproteinase-1 (MT1-MMP) in mammary gland development and tumorigenesis, transgenic mice overexpressing MT1-MMP in mammary gland under the control of the mouse mammary tumor virus long terminal repeat-promoter were generated. The mouse mammary tumor virus/MT1-MMP transgenic mice displayed abnormalities in 82% of female mammary glands. The abnormalities were verified as lymphocytic infiltration, fibrosis, hyperplasia, alveolar structure disruption, dysplasia, and adenocarcinoma. Northern and reverse transcription-PCR analyses demonstrated that MT1-MMP mRNA was overexpressed in mammary glands exhibiting abnormalities. Western blot analysis and immunohistochemical studies have revealed that the protein expression level was also increased in these glands. In addition, the beta-casein gene as a functional epithelial cell marker was poorly expressed in the mammary glands of transgenic mice exhibiting abnormalities. Gelatin zymography showed significantly increased MMP-2 activation in these mammary glands. These results showed that overexpression of MT1-MMP induced remodeling of the extracellular matrix and tumor formation in the mammary glands of transgenic mice. Therefore, we suggest that overexpression of MT1-MMP may play a key role in development and tumorigenesis in mammary glands.

Mucin 1-mediated chemo-resistance in lung cancer cells.

Paclitaxel (PTX) is a commonly used drug to treat diverse cancer types. However, its treatment can generate resistance and the mechanisms of PTX-resistance in lung cancers are still unclear. We demonstrated that non-small cell lung cancers (NSCLCs) survive PTX treatment. Compared with the progenitor NSCLC A549 cells, the PTX-resistant A549 cells (A549/PTX) displayed enhanced sphere-formation ability. The proportion of the cancer stem cell marker, aldehyde dehydrogenase-positive cells, and epithelial-mesenchymal transition signaling protein levels were also elevated in A549/PTX. Importantly, the levels of oncoproteins phosphoinositide-3 kinase/Akt, mucin 1 cytoplasmic domain (MUC1-C) and ß-catenin were also significantly elevated in A549/PTX. Furthermore, nuclear translocation of MUC1-C and ß-catenin increased in A549/PTX. The c-SRC protein, an activator of MUC1-C, was also overexpressed in A549/PTX. These observations led to the hypothesis that enhanced expression of MUC1-C is associated with stemness and PTX resistance in NSCLCs. To test this, we knocked down or overexpressed MUC1-C in A549/PTX and found that inhibition of MUC1-C expression coupled with PTX treatment was sufficient to reduce the sphere-forming ability and survival of A549/PTX. In summary, our in vitro and in vivo studies have revealed a potential mechanism of MUC1-C-mediated PTX resistance and provided insights into a novel therapeutic measure for lung cancers.

Peroxiredoxin II promotes hepatic tumorigenesis through cooperation with Ras/Forkhead box M1 signaling pathway.

The current study was carried out to define the involvement of Peroxiredoxin (Prx) II in progression of hepatocellular carcinoma (HCC) and the underlying molecular mechanism(s). Expression and function of Prx II in HCC was determined using H-ras(G12V)-transformed HCC cells (H-ras(G12V)-HCC cells) and the tumor livers from H-ras(G12V)-transgenic (Tg) mice and HCC patients. Prx II was upregulated in H-ras(G12V)-HCC cells and H-ras(G12V)-Tg mouse tumor livers, the expression pattern of which highly similar to that of forkhead Box M1 (FoxM1). Moreover, either knockdown of FoxM1 or site-directed mutagenesis of FoxM1-binding site of Prx II promoter significantly reduced Prx II levels in H-ras(G12V)-HCC cells, indicating FoxM1 as a direct transcription factor of Prx II in HCC. Interestingly, the null mutation of Prx II markedly decreased the number and size of tumors in H-ras(G12V)-Tg livers. Consistent with this, knockdown of Prx II in H-ras(G12V)-HCC cells reduced the expression of cyclin D1, cell proliferation, anchorage-independent growth and tumor formation in athymic nude mice, whereas overexpression of Prx II increased or aggravated the tumor phenotypes. Importantly, the expression of Prx II was correlated with that of FoxM1 in HCC patients. The activation of extracellular signal-related kinase (ERK) pathway and the expression of FoxM1 and cyclin D1 were highly dependent on Prx II in H-ras(G12V)-HCC cells and H-ras(G12V)-Tg livers. Prx II is FoxM1-dependently-expressed antioxidant in HCC and function as an enhancer of Ras(G12V) oncogenic potential in hepatic tumorigenesis through activation of ERK/FoxM1/cyclin D1 cascade.

Oxygen distribution and consumption within the retina in vascularised and avascular retinas and in animal models of retinal disease.

Maintenance of an adequate oxygen supply to the retina is critical for retinal function. In species with vascularised retinas, such as man, oxygen is delivered to the retina via a combination of the choroidal vascular bed, which lies immediately behind the retina, and the retinal vasculature, which lies within the inner retina. The high-oxygen demands of the retina, and the relatively sparse nature of the retinal vasculature, are thought to contribute to the particular vulnerability of the retina to vascular disease. A large proportion of retinal blindness is associated with diseases having a vascular component, and disrupted oxygen supply to the retina is likely to be a critical factor. Much attention has therefore been directed at determining the intraretinal oxygen environment in healthy and diseased eyes. Measurements of oxygen levels within the retina have largely been restricted to animal studies in which oxygen sensitive microelectrodes can be used to obtain high-resolution measurements of oxygen tension as a function of retinal depth. Such measurements can immediately identify which retinal layers are supplied with oxygen from the different vascular elements. Additionally, in the outer retinal layers, which do not have any intrinsic oxygen sources, the oxygen distribution can be analysed mathematically to quantify the oxygen consumption rate of specific retinal layers. This has revealed a remarkable heterogeneity of oxygen requirements of different components of the outer retina, with the inner segments of the photoreceptors being the dominant oxygen consumers. Since the presence of the retinal vasculature precludes such a simple quantitative analysis of local oxygen consumption within the inner retina, our understanding of the oxygen needs of the inner retinal components is much less complete. Although several lines of evidence suggest that in the more commonly studied species such as cat, pig, and rat, the oxygen demands of the inner retina as a whole is broadly comparable to that of the outer retina, exactly which cell layers within the inner retina have the most stringent oxygen demands is not known. This may be a critical issue if the cell types most at risk from disrupted oxygen supply are to be identified. This paper reviews our current understanding of the oxygen requirements of the inner and outer retina and presents new data and mathematical models which identify three dominant oxygen-consuming layers in the rat retina. These are the inner segments of the photoreceptors, the outer plexiform layer, and the deeper region of the inner plexiform layer. We also address the intriguing question of how the oxygen requirements of the inner retina are met in those species which naturally have a poorly vascularised, or even totally avascular retina. We present measurements of the intraretinal oxygen distribution in two species of laboratory animal possessing such retinas, the rabbit and the guinea pig. The rabbit has a predominantly avascular retina, with only a narrow band of retinal vasculature, and the guinea pig retina is completely avascular. Both these animals demonstrate species adaptations in which the oxygen requirement of their inner retinas are extremely low when compared to that of their outer retinas. This finding both uncovers a remarkable ability of the inner retina in avascular species to function in a low-oxygen environment, and also highlights the dangers of extrapolating findings from avascular retinas to infer metabolic requirements of vascularised retinas. Different species also demonstrate a marked diversity in the manner in which intraretinal oxygen distribution is influenced by increases in systemic oxygen level. In the vascularised rat retina, the inner retinal oxygen increase is muted by a combination of increased oxygen consumption and a reduction of net oxygen delivery from the retinal circulation. The avascular retina of the guinea pig demonstrated a novel and powerful regulatory mechanism that prevents any dramatic rise in choroidal oxygen levels and keeps retinal oxygen levels within the normal physiological range. In contrast, in the avascular regions of the rabbit retina the choroidal oxygen level passively follows the increase in systemic oxygenation, and there is a dramatic rise in oxygen level in all retinal layers. The presence or absence of oxygen-regulating mechanisms may well reflect important survival strategies for the retina which are not yet understood. Intraretinal oxygen measurements in rat models of retinal disease are also presented. We describe how oxygen distribution across the rat retina is influenced by manipulation of systemic blood pressure. We examine the effect of acute and chronic occlusion of the retinal vasculature, and explore the feasibility of meeting the oxygen needs of the ischemic retina from the choroid. (ABSTRACT TRUNCATED)

Characterization of the murine gene encoding 1-Cys peroxiredoxin and identification of highly homologous genes.

A new type of peroxiredoxin, named 1-Cys peroxiredoxin (1-Cys Prx), reduces hydrogen peroxide with the use of electrons from unidentified electron donor(s). We have isolated the mouse gene encoding 1-Cys Prx (CP-3) and shown that it is comprised of five exons and four introns. Analysis of 5' flanking regions revealed binding sequences of several putative transcription factors such as Sp1, Pit-1a, c-Jun, c-Myc and YY1. It is noticeable that several potential Sp1 binding sites assigned the -60 through -96bp from putative transcription initiation site. The gel shift assays showed that Sp1 and Pit-1a bind specifically to each binding site in 1-Cys Prx promoter. We also isolated two highly related genes such as CP-2 and CP-5. These genes are encoded by single exons, and show 85% of nucleotide sequence homology with the CP-3. The structural features of these genes suggest that they might be intronless genes derived from the CP-3 by the mechanism involving retrotransposition. In addition, our data suggest that they are inserted to a specific site of the mouse L1 repetitive element. The 1-Cys Prx was actively transcribed in a variety of adult tissues as well as in the developing embryos. These results suggest that only the 1-Cys Prx gene might be relevant for studying the function of the 1-Cys Prx in the murine system.