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Objective: To assess the overall asthma control level in urban China. Methods: A multi-center, cross-sectional, questionnaire-based survey was carried out in 30 provinces in China (except Tibet). Asthmatic outpatients who met the inclusion criteria were recruited consecutively from 30 sub-centers from October 2015 to May 2016. Data of demographic characteristics, asthma control, asthma management, asthma exacerbations and self-management, and disease perception were collected by face-to-face interviews. Results: Totally 3 875 asthmatic outpatients were recruited. Asthma control level was assessed according to GINA 2015. The prevalence of asthma control was 28.5%(1 099/3 854). Among 10 cities, which were also involved in the asthma control survey in 2008, the level of asthma control improved from 28.7% in 2008 to 39.2%(533/1 361) in 2016. The rate of hospitalizations due to asthma exacerbations was 26.4%(1 017/3 858) and the rate of emergency visits was 22.4%(864/3 858). Among adult patients, 18.4%(568/3 092) of them had been absent from work while 63.6%(63/99) adolescent patients had been absent from school because of asthma attacks in the previous year. Conclusion: Although the level of asthma control in China is still far from ideal at present, the status has been improved in some cities. Poorly controlled asthma resulted in increase of hospitalizations, emergency department visits and work/school absences.

Objective: To investigate the current perception of disease among asthmatic patients in Chinese urban areas, and to address its association with asthma control. Methods: This was a nationwide, multi-center, cross-sectional study covering 30 third-level, grade A hospitals in 30 provinces, municipalities and autonomous regions (not including Tibet) across Chinese mainland. The survey was carried out from October 2015 to May 2016. Asthmatic outpatients were selected to receive face-to-face questionnaire survey on asthma control including Asthma Control Test (ACT) score, classification of asthma control level[according to the Global Initiative for Asthma (GINA) 2015 classification system, as assessed by the physician completing the survey], perception of asthma (including question 1: nature of asthma as a disease; question 2: selection of first-line therapeutic agents to be regularly used daily for asthma; question 3: appropriate timing of the use of short-acting aerosols of β(2) receptor agonists; and question 4: therapeutic goals for asthma). Results: A total of 3 875 asthmatic outpatients were included; among them, 69.0% (2 660/3 857) were aware that asthma is "an airway inflammatory disorder resulting from the combined effects of heredity and environment" ; 60.2% (2 321/3 857) considered "inhaled glucocorticoids or their compound preparations" to be the first-line therapeutic agents to be regularly used daily for patients with persistent chronic asthma; 85.7% (3 277/3 823) considered it appropriate to use short-acting aerosols of β(2) receptor agonists "as needed in the event of disease aggravation or acute exacerbation" ; and 75.4% (2 761/3 661) were aware that asthma "can be adequately or completely controlled in the long term" . The ACT score[20 (16, 23) vs 19 (16, 22) points; Z =-3.928, P <0.001]and asthma control rate (29.92% vs 25.31%; χ(2)=8.616, P =0.003) were significantly higher, and the rate of uncontrolled asthma (19.92% vs 23.48%; χ(2)=6.267, P =0.012) was significantly lower among the 2 660 (69.0%) patients correctly answering question 1 than among the 1 197 (31.0%) patients giving incorrect answer. The ACT score[21 (17, 23) vs 19 (15, 22) points; Z =-9.190, P <0.001] and asthma control rate (32.66% vs 22.20%; χ(2)=49.614, P <0.001) were significantly higher, and the rate of uncontrolled asthma (18.40% vs 25.00%; χ(2)=24.267, P <0.001) was significantly lower among the 2 321 (60.2%) patient correctly answering question 2 than among the 1 536 (39.8%) patients giving incorrect answer. Conclusions: Compared to previous surveys, there has been improved perception of disease among asthmatic patients in Chinese urban areas. Correct perception of disease is favorable for improving asthma control level.

An interface electron state at the junction between a three-dimensional topological insulator film, Bi 2 Se 3 , and a ferrimagnetic insulator film, Y 3 Fe 5 O 12 (YIG), was investigated by measurements of angle-resolved photoelectron spectroscopy and x-ray absorption magnetic circular dichroism. The surface state of the Bi 2 Se 3 film was directly observed and localized 3d spin states of the Fe 3+ in the YIG film were confirmed. The proximity effect is likely described in terms of the exchange interaction between the localized Fe 3d electrons in the YIG film and delocalized electrons of the surface and bulk states in the Bi 2 Se 3 film.

We investigated the mechanisms mediating hepatic metabolic responses to an acute lipopolysaccharide (LPS) challenge in goats. Guanzhong dairy goats (15) were randomly divided into three groups: control (CTL, saline, 0.2 ml/kg BW), lower dose LPS (LPS-L, 20 μg/kg BW) and higher dose LPS (LPS-H, 40 μg/kg BW). All injections were administered intraperitoneally twice with a 24-h interval. Forty-eight hours after the first injection, blood samples were collected to extract plasma for biochemical analysis, and liver tissues were biopsied and stored in liquid nitrogen for metabonomics analysis. We found that plasma levels of alanine aminotransferase, aspartate aminotransferase and total bilirubin increased (p < 0.05) in both LPS-treated groups, whereas plasma triglyceride, cholesterol, very low-density lipoprotein, low-density lipoprotein, high-density lipoprotein, total protein and albumin levels markedly decreased (p < 0.05). The increased activities of alanine aminotransferase (ALT) and aspartate aminotransferase (AST), levels of tumour necrosis factor α (TNF-α), interleukin (IL)-1β, IL-6 and IL-8 indicated hepatic injury and metabolic dysfunction in some degree. Using proton nuclear magnetic resonance ( 1 H-NMR) metabonomics and the Chenomx NMR suite database, 69 metabolites were detected and identified. Metabolic differences among the groups were determined with pattern recognition analyses using principal component analysis and supervised projection to latent structures discriminant analysis. Pattern recognition analysis distinguished and clustered the metabolite variables from the three groups, finding nine of 69 metabolites that differed significantly between two of the three groups: six from the LPS-L or LPS-H groups differed from CTL and three differed between LPS-L and LPS-H groups. These altered metabolites were closely connected with glucose, lipid and amino acid metabolic pathways in hepatocytes. Based on an analysis of these metabolites and their relevant pathways, the mechanisms and degree of hepatic injury were deduced. Therefore, the metabolic profile was used effectively to detect characteristic hepatic metabolites, discriminate metabolic changes induced by LPS, clarify the mechanisms for the resulting metabolic dysfunctions and provide efficient information to diagnose liver injury., (Journal of Animal Physiology and Animal Nutrition © 2016 Blackwell Verlag GmbH.)

Objective: To analyze the correlation between white matter integrity and spatial navigation impairment in patients with mild cognitive impairment (MCI). Methods: A total of 27 MCI subjects and 24 healthy controls were enrolled from the Affiliated Drum Tower Hospital of Nanjing University Medical School from May 2015 to February 2016, who underwent 3.0 T MRI scan and 2D-computer version spatial navigation test.DTI preprocessing and tract-based spatial statistics (TBSS) were performed by PANDA.Two sample t -test and partial correlation coefficients were performed to investigate the correlation of white matter impairments and spatial navigation decline. Results: Relative to controls, MCI showed worse egocentric navigation ( t =-2.202, P <0.05). Decreased FA in superior longitudinal fasciculus (left t =2.95, right t =2.95, P <0.05), inferior longitudinal fasciculus and inferior fronto-occipital fasciculus (left t =2.66, right t =2.96, P <0.05), corpus callosum ( t =2.09, P <0.05), cingulum (left t =2.76, right t =2.41, P <0.05), fornix ( t =4.83, P <0.05), and corticospinal tract (left t =2.33, right t =2.26, P <0.05), were found in the MCI subjects.The decreased FA value of superior longitudinal fasciculus (left r =-0.354, right r =-0.347, P <0.05), inferior longitudinal fasciculus (left r =-0.338, right r =-0.336, P <0.05), cingulum (left r =-0.395, right r =-0.370, P <0.05), right corticospinal tract ( r =-0.362, P <0.05) and fornix ( r =-0.369, P <0.05) were correlated with increased ego average total error.Allo average total error were negative correlated with FA value of superior longitudinal fasciculus (left r =-0.329, right r =-0.350, P <0.05), inferior longitudinal fasciculus (left r =-0.349, right r =-0.378, P <0.05), splenium of corpus callosum ( r =-0.364, P <0.05) and cingulum (left r =-0.340, right r =-0.406, P <0.05). Conclusion: This study implicated the potential white matter structural basis of spatial navigation impairment and will have an impact on the further study of the neurobiological mechanisms of human spatial navigation ability.

Objective: To determine the relevance between the SOX 10 mutation and Waardenburg syndrome (WS) accompanied with inner ear abnormality by analyzing the inner ear imaging results and molecular and genetic results of the WS patients with the SOX 10 mutation. Methods: This study included 36 WS in patients during 2001 and 2015 in the department of otorhinolaryngology head and neck surgery, Chinese Peoples's Liberation Army General Hospital. The condition of the inner ear of each patient was assessed by analyzing HRCT scans of the temporal bone and MRI scans of the brain and internal auditory canal. Meanwhile, the possible pathogenic genes of WS, including SOX10, MITF , and PAX 3, were also screened. Patients were divided into two groups according to SOX 10 mutation.The Fisher accuracy test was used to determine statistical difference of inner ear deformation incidence between the two groups. Results: Among all 36 patients, 12 were found to have inner ear abnormality. Most abnormalities were posterior semicircular canal deformations, some accompanied with cochlear deformation and an enlarged vestibule. Among all patients, 9 patients were SOX 10 heterozygous mutation carriers, among which six showed bilateral inner ear abnormality. Fisher accuracy test results suggested a significant correlation between the SOX 10 mutation and inner ear abnormality in WS patients ( P =0.036). Conclusion: This study found that WS patients with the SOX 10 mutation are more likely to have deformed inner ears when compared to WS patients without the SOX 10 mutation.

Objective: To analyze the etiology and clinical symptoms and to investigate the therapeutic strategies of cerebrospinal fluid otorrhea. Method: A retrospective analysis of 37 cases of patients with cerebrospinal fluid otorrhea.The clinical symptoms, auxiliary examination, intraoperative findings, surgical methods and postoperative follow-up were analyzed. Result: In 37 cases, 35 patients underwent the plugging surgery once and cured, 1 patient with inner ear malformation underwent another operation and cured, 1 patient didn't have the operation. No cerebrospinal fluid leakage or meningitis recurrence was reported by the followed up from 1 months to 7 years after operation. Conclusion: Surgical repair is an effective method to treat the cerebrospinal fluid otorrhea. It is significant to take appropriate surgical approach to expose and to find the leak, according to the etiological factor and imaging examination., Competing Interests: The authors of this article and the planning committee members and staff have no relevant financial relationships with commercial interests to disclose., (Copyright© by the Editorial Department of Journal of Clinical Otorhinolaryngology Head and Neck Surgery.)

Objective: To investigate the effect of Toll-like receptor 2 (TLR2) on the inhibition role of sevoflurane on airway inflammation in asthmatic mice., Methods: The lung tissue samples of C(57) BL/6 mice used in this study were from previous research, including control group, asthma group and sevoflurane group, 8 samples in each group. Twenty-four specific pathogen free female TLR2 gene deletion (TLR2(-/-)) mice were randomly assigned to control group, asthma group and sevoflurane group, with 8 mice in each group. Asthma group and sevoflurane group were then sensitized and challenged with ovalbumin (OVA) to establish asthma model, combined with repeated inhalation of 3% sevoflurane in sevoflurane group. In C(57) mice, expression levels of TLR2 were detected using Western blotting analyses. In TLR2(-/-) mice, numbers of differential inflammatory cells were investigated; levels of tumor necrosis factor-alpha (TNF-α) and interleukin-10 (IL-10) in bronchoalveolar lavage fluid (BALF) were measured by enzyme linked immunosorbent assay (ELISA); lung tissue inflammation was detected with HE staining., Results: In lung tissues from C(57) mice, levels of protein expression of TLR2 in asthma group (0.547±0.042) were higher than those in control group (0.312±0.023) (P=0.023) and sevoflurane group (0.287±0.033) (P=0.020). In TLR2(-/-) mice, the number of total cells ((83.13±19.43)×10(3)/ml), numbers of differential inflammatory cells and TNF-α level ((546±16) pg/ml) in BALF in sevoflurane group were lower than those in asthma group ((206.43±41.82)×10(3)/ml, (732±41) pg/ml), but still higher than those in control group ((44.64±7.17)×10(3)/ml, (380±24) pg/ml) (all P<0.05); lung tissue inflammation was inhibited in sevoflurane group than in asthma group, but still more obvious than that in control group., Conclusion: Toll like receptor 2 involved in the anti-inflammatory effect of sevoflurane on asthmatic airway inflammation in mice.

Several reports indicate that melatonin alleviates bleomycin (BLM)-induced pulmonary fibrosis in rodent animals. Nevertheless, the exact mechanism remains obscure. The present study investigated the effects of melatonin on endoplasmic reticulum (ER) stress and epithelial-mesenchymal transition (EMT) during BLM-induced lung fibrosis. For the induction of pulmonary fibrosis, mice were intratracheally injected with a single dose of BLM (5.0 mg/kg). Some mice were intraperitoneally injected with melatonin (5 mg/kg) daily for a period of 3 wk. Twenty-one days after BLM injection, lung fibrosis was evaluated. As expected, melatonin significantly alleviated BLM-induced pulmonary fibrosis, as evidenced by Sirius red staining. Moreover, melatonin significantly attenuated BLM-induced EMT to myofibroblasts, as determined by its repression of α-SMA expression. Further analysis showed that melatonin markedly attenuated BLM-induced GRP78 up-regulation and elevation of the cleaved ATF6 in the lungs. Moreover, melatonin obviously attenuated BLM-induced activation of pulmonary eIF2α, a downstream target of the PERK pathway. Finally, melatonin repressed BLM-induced pulmonary IRE1α phosphorylation. Correspondingly, melatonin inhibited BLM-induced activation of XBP-1 and JNK, two downstream targets of the IRE1 pathway. Taken together, these results suggest that melatonin alleviates ER stress and ER stress-mediated EMT in the process of BLM-induced pulmonary fibrosis.

Apolipoprotein A5 (APOA5) and C3 (APOC3) genes are involved in the PPAR lipid metabolism pathway and thus associated with elevated triglyceride levels. However, whether APOA5 and APOC3 genetic polymorphisms affect intramuscular fat deposition and other meat quality traits remains unknown in pigs. One hundred and seventy-one Kele pigs were sampled to investigate genetic variants in the APOA5 and APOC3 genes and their association with seven pork quality traits. We identified 5 single nucleotide polymorphisms (SNPs) in the promoter region of the APOA5 gene and 17 SNPs in the APOC3 gene. Linkage disequilibrium analysis revealed 5 complete linkage disequilibria among these 22 SNPs. We found that 10 SNPs were significantly correlated with meat quality traits, including the mutation A5/-769 in the APOA5 gene, which was significantly associated with cooked weight percentage, and 9 SNPs in the APOC3 gene that were significantly associated with drip loss rate, meat color value of longissimus dorsi muscle and shear force. Therefore, these SNP markers will be useful for marker-assisted selection for improved pork quality.

Recent advances in defining neurotrophin signaling mediators have provided insights into the signal transduction mechanisms that underlie axon growth. Evidence is accumulating that major Trk effectors regulate the morphological development of embryonic peripheral neurons. Less is known about signaling related to the robust axon extension that follows peripheral axotomy of adult neurons. Regenerative axon growth can be mimicked in vitro by a "conditioning" lesion performed 2 weeks before culture (Smith and Skene, 1997). Previous work has implicated both neurotrophins and cytokines in this response. Because signal transduction mediators of both of these families of growth factors are well characterized, we have compared the role of neurotrophin and cytokine signaling in developmental versus regenerative sensory axon growth. Chemical inhibitors were administrated to embryonic and axotomized sensory neurons in vitro to block the activation of Erk kinase (MEK)-extracellular signal-regulated kinase (ERK), phosphatidylinositol-3 kinase (PI3-K), and janus kinase (JAK) signaling. As expected, both MEK and PI3-K inhibition blocked axon growth from both naive and NGF-stimulated embryonic day 13 sensory neurons, whereas inhibition of JAK phosphorylation had no effect. In contrast, neither MEK nor PI3-K inhibitors blocked elongation of adult sensory neurons after a conditioning lesion. However, the addition of a JAK2 inhibitor prevented the regenerative axon response. Consistent with these pharmacological results, the percentage of neurons showing intense nuclear signal transducers and activators of transcription 3 phosphorylation after a conditioning lesion was markedly increased compared with controls. These observations demonstrate that the signaling mediators that underlie regenerative axon growth are distinct from those used during development and suggest that cytokine signaling may be critical to peripheral nervous system regeneration.

A world-class lightweight rower who came to the Hong Kong Sports Institute was evaluated over a two-month period of training. He was preparing for the 1995 World Rowing Championships. The subjects trained for approximately 70 minutes per day at different levels of intensity, to simulate race conditions. Caloric expenditures (based upon heart rate response and estimation from specific activity) were estimated to be approximately 4125 kcal/day. A three day dietary survey revealed that the subject was able to maintain caloric balance (caloric intake = 4088 kcal/day). Body weight was maintained at a constant level, another indication that the subject was able to meet energy needs. A breakdown of his food intake revealed protein, carbohydrate, and fat, made up 19%, 51%, and 30% of his caloric intake, respectively. This was somewhat surprising since recommendations for endurance athletes are for higher carbohydrate contribution. During and after the training bouts the subject consumed two separate carbohydrate supplements. A total of 218 g of carbohydrate were consumed daily. This level of supplementation was able to maintain the subject's blood glucose concentration at normal levels following recovery from exercise. Comparisons of recovery heart rates when either water or carbohydrate was consumed indicate a beneficial effect of the supplement. Despite consuming carbohydrates at lower than recommended levels, this athlete went on to win this third gold medal in rowing.

To elucidate mechanisms of tumor necrosis factor alpha (TNF-alpha)-induced proliferation of a number of human leukemia and lymphoma cell lines, we examined the role of p38 mitogen-activated protein kinase (MAPK) in TNF-alpha signaling in Mo7e and Hut-78 cells. TNF-alpha-dependent p38 MAPK activation was detected in both Mo7e and Hut-78 cells and was blocked by the p38 MAPK inhibitor, SB203580. Ablation of p38 MAPK activity by SB203580 abrogated TNF-alpha-induced Mo7e cell proliferation and TNF-alpha-dependent autocrine growth of Hut-78. As we have shown previously that activation of the nuclear factor kappaB (NF-kappaB) is also required for TNF-alpha-induced Mo7e cell proliferation, the involvement of p38 MAPK in NF-kappaB activation was assessed. SB203580 did not affect TNF-alpha-signaled nuclear translocation and DNA-binding activity of NF-kappaB, and inhibition of NF-kappaB function did not affect TNF-alpha-induced p38 MAPK activation, indicating that these events are not dependent on each other. However, SB203580 depressed the expression of NF-kappaB-dependent genes, as monitored by a kappaB-driven reporter gene. Our findings demonstrate that activation of both p38 MAPK and NF-kappaB plays a critical role in TNF-alpha-mediated survival and proliferation of human leukemia and lymphoma cells, and p38 MAPK acts at least in part by facilitating the transcriptional activation function of NF-kappaB.

Circadian rhythm disturbances are frequently present in Alzheimer disease (AD). In the present study, we investigated the expression of vasopressin (AVP) mRNA in the human suprachiasmatic nucleus (SCN). The in situ hybridization procedure on formalin-fixed paraffin-embedded material was improved to such a degree that we could, for the first time, visualize AVP mRNA expressing neurons in the human SCN and carry out quantitative measurements. The total amount of AVP mRNA expressed as masked silver grains in the SCN was 3 times lower in AD patients (n = 14; 2,135 +/- 597 microm2) than in age- and time-of-death-matched controls (n = 11; 6,667 +/- 1466 microm2) (p = 0.003). No significant difference was found in the amount of AVP mRNA between AD patients with depression (n = 7) and without depression (n = 7) (2,985 +/-1103 microm2 and 1,285 +/- 298 microm2, respectively; p = 0.38). In addition, the human SCN AVP mRNA expressing neurons showed a marked day-night difference in controls under 80 years of age. The amount of AVP mRNA was more than 3 times higher during the daytime (9,028 +/- 1709 microm2, n = 7) than at night (2,536 +/- 740 microm2, n = 4; p = 0.02), whereas no clear diurnal rhythm of AVP mRNA in the SCN was observed in AD patients. There was no relationship between the amount of AVP mRNA in the SCN and age at onset of dementia, duration of AD and the neuropathological changes in the cerebral cortex. These findings suggest that the neurobiological basis of the circadian rhythm disturbances that are responsible for behavioral rhythm disorders is located in the SCN. It also explains the beneficial effects of light therapy on nightly restlessness in AD patients.

The factor-independent Dami/HEL and Meg-01 and factor-dependent Mo7e leukemic cell lines were used as models to investigate JAK/STAT signal transduction pathways in leukemic cell proliferation. Although Dami/HEL and Meg-01 cell proliferation in vitro was independent of and unresponsive to exogenous cytokines including granulocyte-macrophage colony-stimulating factor (GM-CSF), interleukin-3 (IL-3), IL-6, thrombopoietin (TPO), and tumor necrosis factor-alpha (TNF-alpha), the growth of Mo7e cells was dependent on hematopoietic growth factors. When these cell lines were cultured in medium without cytokines, a constitutively activated STAT-like DNA-binding factor was detected in nuclear extracts from both Dami/HEL and Meg-01 cells. However, the STAT-like factor was not detectable in untreated Mo7e cells, but was activated transiently in Mo7e cells in response to cytokine treatments. The constitutively activated and cytokine-induced STAT-like DNA-binding factor in these three cell lines was identified as STAT5 by oligonucleotide competition gel mobility assays and by specific anti-STAT antibody gel supershift assays. Constitutive activation of JAK2 also was detected in the factor-independent cell lines, but not in Mo7e cells without cytokine exposure. Meg-01 cells express a p185 BCR/ABL oncogene, which may be responsible for the constitutive activation of STAT5. Dami/HEL cells do not express the BCR/ABL oncogene, but increased constitutive phosphorylation of Raf-1 oncoprotein was detected. In cytokine bioassays using growth factor-dependent Mo7e and TF-1 cells as targets, conditioned media from Dami/HEL and Meg-01 cells did not show stimulatory effects on cell proliferation. Our results indicate that the constitutive activation of JAK2/STAT5 correlates with the factor-independent growth of Dami/HEL and Meg-01 cells. The constitutive activation of JAK2/STAT5 in Dami/HEL cells is triggered by a mechanism other than autocrine cytokines or the BCR/ABL oncoprotein.

Sleep disruption, nightly restlessness, sundowning, and other circadian disturbances are frequently seen in Alzheimer's disease (AD) patients. Changes in the suprachiasmatic nucleus and pineal gland are thought to be the biological basis for these behavioral disturbances. Melatonin is the main endocrine message for circadian rhythmicity from the pineal. To determine whether melatonin production was affected in AD, melatonin levels were determined in the cerebrospinal fluid (CSF) of 85 patients with AD (mean age, 75 +/- 1.1 yr) and in 82 age-matched controls (mean age, 76 +/- 1.4 yr). Ventricular postmortem CSF was collected from clinically and neuropathologically well defined AD patients and from control subjects without primary neurological or psychiatric disease. In old control subjects (>80 yr of age), CSF melatonin levels were half of those in control subjects of 41-80 yr of age [176 +/- 58 (n = 29) and 330 +/- 66 (n = 53) pg/mL, respectively; P = 0.016]. We did not find a diurnal rhythm in CSF melatonin levels in control subjects. In AD patients the CSF melatonin levels were only one fifth (55 +/- 7 pg/mL) of those in control subjects (273 +/- 47 pg/mL; P = 0.0001). There was no difference in the CSF melatonin levels between the presenile (42 +/- 11 pg/mL; n = 21) and the senile (59 +/- 8 pg/mL; n = 64; P = 0.35) AD patients. The melatonin level in AD patients expressing apolipoprotein E-epsilon3/4 (71 +/- 11 pg/mL) was significantly higher than that in patients expressing apolipoprotein E-epsilon4/4 (32 +/- 8 pg/ml; P = 0.02). In the AD patients no significant correlation was observed between age of onset or duration of AD and CSF melatonin levels. In the present study, a dramatic decrease in the CSF melatonin levels was found in old control subjects and even more so in AD patients. Whether supplementation of melatonin may indeed improve behavioral disturbances in AD patients should be investigated.

We investigated the effects of tumor necrosis factor (TNF) alpha on the human megakaryocytic leukemic cell lines Mo7e, Meg-01, and Dami/HEL. Our data show that both type I and type II TNF receptors (TNF-RI and TNF-RII) are expressed on all of these cells, and TNF-alpha significantly stimulates the proliferation of growth factor-dependent Mo7e cells but not of Meg-01 or Dami/HEL cells, which grow in a factor-independent manner. TNF-alpha serves predominantly as a mitogen for Mo7e cell proliferation and does not induce Mo7e cell differentiation. Coincubation with both TNF-alpha and anti-TNF-alpha neutralizing antibody completely abolishes the TNF-alpha-induced proliferation of Mo7e cells. In bioassays, there is no detectable level of other stimulatory cytokines in conditioned medium from Mo7e cells previously stimulated by TNF-alpha, implying that the stimulatory effect of TNF-alpha on Mo7e cells is derived from the direct action of TNF-alpha rather than via the induction of secondary cytokines by TNF-alpha. Flow cytometric studies demonstrated that TNF-alpha binds to Mo7e cells that have been pretreated with either anti-TNF-RI or anti-TNF-RII neutralizing antibody, but TNF-alpha does not bind to cells pre-exposed to both receptor antibodies. However, the incubation of Mo7e cells with either TNF-RI or TNF-RII neutralizing antibodies or with either soluble TNF-RI or TNF-RII inhibits TNF-alpha-induced cell proliferation, indicating the requirement of interactions with both TNF receptors for the mitogenic activity of TNF-alpha. Furthermore, our data suggest that an alternative signaling pathway may be involved in TNF-alpha-induced Mo7e cell proliferation, because the common mitogen-activated protein kinase (MAPK) and signal transducer and activator of transcription (STAT) signaling pathways activated by other cytokines that induce Mo7e cell proliferation are not activated by TNF-alpha.

The proliferative potential of cultured rat glioma cells (C6 and 9L) was evaluated after hyperthermia using immunohistochemical staining with bromodeoxyuridine (BrdU) and Ki-67 monoclonal antibodies. Apoptosis was assessed by in situ end-labeling of deoxyribonucleic acid breaks. Both BrdU and Ki-67 labeling indexes decreased with increasing hyperthermia time. The decrease of the Ki-67 labeling index was not as great as that of the BrdU labeling index. The number of apoptotic cells increased with time after hyperthermia. These results indicate that the antitumor effect of hyperthermia may reflect the induction of apoptosis in the cells within the cell cycle, and the resultant reduction of the proliferative potential of surviving cells, especially in the S phase.

A 50-year-old male presented with a rare intraparenchymal metastatic tumor spreading through the periventricular tissue. Magnetic resonance (MR) imaging demonstrated the tumor as a heterogeneous low-intensity area on T1-weighted images with enhancement by gadolinium-diethylenetriaminepenta-acetic acid, and as a heterogeneous high- or isointensity area on T2-weighted images. Histological examination of a biopsy sample showed adenocarcinoma. This MR imaging appearance is typical of malignant glioma. The differential diagnosis of tumor in the cerebral parenchyma with ventricular dissemination should include both primary and secondary intracranial malignant tumors. MR imaging is useful in the diagnosis of such tumors, but the final diagnosis should be based on either tissue biopsy or cytological examination of the cerebrospinal fluid.

It was reported that chemical stresses such as arsenite, cadmium or salicylate fail to induce synthesis of the inducible form of HSP70 (HSP70i). We report here that exposure of cells to higher doses of these chemical treatments induced significant synthesis of HSP70i in CHO cells as well as other cell lines. The synthesis of HSP70i is primarily regulated at the transcriptional level. Although all tested chemical treatments induced heat shock factor (HSF) binding to the heat shock element (HSE), HSP70i synthesis appears to be regulated by an alternative factor (CHBF) which constitutively binds to the HSE at 37 degrees C. The treatments, which dissociate the HSE-CHBF complex, induced significant HSP70i synthesis. The treatments, which failed to induce HSP70i synthesis, still activated HSF binding to HSE but the HSE-CHBF complex remained as that of untreated control cells.

The effects of stimulation of the lateral reticular nucleus (LRN) on responses of spinal dorsal horn neurons produced by formalin (5%, 50 microliters, sc) were investigated. In 12 out of 19 neurons tested, formalin induced 2 distinct phases of response. The first phase was initiated immediately after the injection of formalin and lasted for 3-8 min. The second phase started gradually 20-35 min after administration of formalin and lasted 30-65 min. Both phases of the response were inhibited by LRN stimulation.

We report here our studies on the inhibitional effect of 45 degrees C heat shock on transcriptional and translational activity and their subsequent recovery at 37 degrees C in Rat-1 cells, thermotolerant Rat-1 cells (TT Rat-1), and Rat-1 cells transfected with human hsp70 gene (HR24, M21). Specifically, we ask whether overexpression of hsp70 protects cells from heat-induced inhibition in RNA and protein synthesis, and/or facilitates cells' ability to recover from translational or transcriptional inhibition after heat shock treatment. Our data demonstrate that the constitutive expression of human hsp70, by itself, confers thermal resistance as expressed in enhanced survival and translational tolerance, but not transcriptional tolerance. In addition, the expression of human hsp70 in Rat-1 cells facilitates cells' ability to recover from heat-induced inhibition in protein and RNA synthesis. After healing at 45 degrees C for 25 min, the time required for RNA and protein synthesis to recover is considerably shorter in HR24, M21, and TT Rat-1 cells than that in control Rat-1 cells. These results provide strong evidence for a direct link between the expression of a functional form of mammalian hsp70, and cells' translational tolerance, as well as their ability to recover from translational and transcriptional inhibition after heat shock.

The major heat-shock protein, hsp-70, is synthesized by cells from a wide variety of organisms in response to heat shock or other stresses. It is assumed that hsp-70 may have an important thermal protective function. To test this hypothesis directly, we have transfected rat fibroblast cells with appropriate expression plasmids containing a cloned human hsp-70 gene. Stable transfectants expressing the human hsp-70 gene product were identified by Western blot analysis. During the course of selecting successful transfectants, we found that when standard methods of sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and immunostaining were employed with the monoclonal antibody raised against the human hsp-70 antigen, we were unable to differentiate the human hsp-70 from the heat-inducible rat hsp-70. This was because the monoclonal antibody cross-reacts with the human and rat proteins, which have the same mobility in SDS-PAGE, and it is difficult to determine which protein is expressed. To improve the resolution of the Western blot technique, we performed additional immunoblot analysis of cellular proteins separated by slab gel isoelectrofocusing. Our study shows that the isoelectrofocusing technique, when combined with antibodies against hsp-70, gave a better resolution for the separation of exogenous human hsp-70 and the endogenous rat hsp-70 than the commonly used SDS-PAGE Western blot analysis. It provides a rapid and specific method to identify positive colonies that express the human hsp-70 gene in transfected rodent cells.

A series of short AVP analogs with D- or L-arginine forming the C-terminal was synthesized, and their peripheral effects, i.e., vasoconstrictor and antidiuretic activities, and behavioral effects in enhancing retention in passive avoidance in rats were evaluated. We found that: 1) AVP(4-8) and its cysteinyl methyl ester were more potent in the behavioral response than its D-Arg homologs; 2) the methyl ester derivative was the most effective analog in this behavioral test among the peptides we synthesized, with a potency 40 times as high as AVP; 3) neither the D- nor the L-Arg short derivatives of AVP showed any peripheral effects up to a dose thousands of times greater than AVP. The results support the contention that arginine in the short analogs plays an important role in their behavioral response associated with a relatively steady peptide conformation.

Adult rats were trained in a maze for brightness discrimination (BD) or in a shuttle box for active avoidance. It was found that the pretreatment of 1-desamino-8-D-arginine-vasopressin (DDAVP) enhanced the retention of brightness discrimination and retarted the extinction of avoidance response. Neonatal daily administration for 14 d of both 9-desglycylamide-DDAVP (DGDDAVP) or 2-destyrosyl-3-desphenylalanyl-9-desglycylamide-AVP (AVP4-8), but not D-arginine-AVP4-8 (DAVP4-8) showed significant facilitation on acquisition and subsequent maintenance of BD in adult rats. The results indicate that some of synthetic arginine-vasopressin analogs may directly act on CNS and consequently affect learning behaviors in rats.

For a distribution F on R(p) and a point x in R(p) the simplicial depth D(x), which is the probability that x be inside a random simplex whose vertices are p + 1 independent observations from F, is introduced. D(x) can be viewed as a measure of depth of the point x relative to F, and its empirical version gives rise to a natural ordering of the data points from the center outward. This ordering provides an approach for detecting outliers in a multivariate data cloud and leads to the introduction of affine equivariant multivariate generalizations of the univariate sample median and L-statistics. This sample median is shown to be consistent for the center of any angularly symmetric distribution.

Groups of newborn Wistar rats received daily 1-desamino-8-D-arginine-vasopressin (DDAVP), oxytocin (OXT), hypertonic saline or normal saline for 14 days from day 1 to day 14 of life. One or three months later they were trained in a maze for brightness discrimination (BD). A group of untreated adult male rats received posttrial DDAVP or normal saline for brightness discrimination. Subsequently all the retentions of BD were tested after one month. We found that the neonatal treatments with both DDAVP and hypertonic saline facilitated acquisition and subsequent maintenance of brightness discrimination in immature and mature rats, and also that posttreatment with DDAVP enhanced retention of BD in adult rats. Oxytocin and normal saline had no effect on these parameters. The results are interpreted as showing that endogenous AVP and its synthetic analog enhance the development and adult function of central neural substrates involved in learning behaviors.

A full-length human placental aromatase cDNA clone, Aro 2, was isolated upon screening a human placental cDNA library with an aromatase cDNA probe and an oligonucleotide probe whose sequence was derived from a human aromatase genomic clone. Nucleotide sequence microheterogeneity was found in the 3'-untranslated region among Aro 2 and in two previously described human aromatase cDNA clones. Both the minor sequence differences and the expression of a single protein species in placental tissue suggest the presence of different alleles for aromatase. Northern blot analyses using one cDNA and two oligonucleotide probes are consistent with the two mRNA messages of 2.9 and 2.5 kilobases arising in human placenta as a consequence of differential processing. Several yeast expression plasmids containing the aromatase cDNA we cloned were constructed. The enzyme was expressed in Saccharomyces cerevisiae. The expressed activity was inhibited by the known aromatase inhibitor, 4-hydroxyandrostenedione. A level of 2 micrograms aromatase/mg partially purified yeast microsomes was estimated by analyses of carbon monoxide difference spectra on microsomal fractions from yeast carrying plasmid pHARK/VGAL. Using [1 beta, 2 beta-3H]androst-4-ene-3,17-dione as the substrate, an apparent Michaels-Menken constant (Km) of 34 nM and a maximum velocity (Vmax) of 23 pmol [3H]water formed per min/mg protein were obtained for the yeast synthesized aromatase by transformation with plasmid pHARK/VGAL. The kinetic results are similar to those determined for human placental aromatase, and suggest that the yeast synthesized aromatase will be useful for further structure-function studies.