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Objective: Adult stem cell senescence and exhaustion are important drivers of organismal age. Restored stem cell self-renewal has revealed novel therapeutic targets for decreasing the incidence of age-associated diseases (AADs) and prolonging the human health span. Transient ectopic expression of the reprogramming factors Oct3/4, Sox2, Klf4 and c-Myc (collectively known as OSKM) in somatic cells can induce partial cellular reprogramming and effectively ameliorate their age-associated hallmarks. However, how this form of rejuvenation is applied to senescent stem cells remains unknown., Materials and Methods: The Integrin-α6highCD71high epidermal stem cells (ESCs) with low self-renewal ability were sorted by flow cytometry and then treated by the interrupted reprogramming induced by transient expression of OSKM. The ability of secondary clones' generation and self-proliferation in vitro, as well as stem cell marker p63, were detected to determine their self-renewal ability. Besides, gene and protein of epidermal cell markers were detected to determine whether their cell identities were retained. Finally, DNA methylation age (eAge) and DNA dehydroxymethylase/methyltransferase were analyzed to explore the alternation of their global DNA methylation pattern during this rejuvenation., Results: The partial reprogramming restored the youthful self-renewal and proliferation in senescent ESCs, including larger secondary clone generation, higher expression of stem cell marker p63 and proliferation marker Ki67, and faster proliferation speed, in each case without abolishing epithelial cellular identity. Moreover, the rejuvenation of adult stem cells could be maintained for 2 weeks after reprogramming factor withdrawal, which was more stable than that of differentiated somatic cells. Additionally, we found that partial reprogramming counteracted the acceleration of eAge in senescent epidermal stem cells and DNA methyltransferase 1 (DNMT1) may play a crucial role in this process., Conclusions: Partial reprogramming has high therapeutic potential for reversing adult stem cell age, providing an advanced way to treat AADs.

Objective: To analyze the difference between Z score and previous criteria in the diagnosis characteristics of coronary artery aneurysm (CAA) in Kawasaki disease, and to investigate the clinical distribution of Kawasaki disease CAA in the Z score group. Methods: This study retrospectively analyzed the clinical and echocardiographic data of 2 419 children with Kawasaki disease in Shenzhen Children's Hospital from January 2009 to December 2019. The traditional criteria and Z score criteria were used to diagnose CAA, and the differences of diagnostic efficiency between the 2 diagnostic methods were analyzed. The clinical distribution characteristics of CAA in children with Kawasaki disease were analyzed by grouping their sex, clinical classification (complete Kawasaki disease, incomplete Kawasaki disease) the sensitivity to intravenous immunoglobulin (IVIG) (IVIG-sensitive Kawasaki disease,IVIG-unresponsive Kawasaki disease). And the course of the disease (≤6 weeks, >6-8 weeks, >8 weeks to 6 months) etc. The χ² test or Kruskal-Wallis test was used for comparison between the groups, and the Kappa test was used for consistency evaluation. Results: Among the 2 419 children with Kawasaki disease, 1 558 were males and 861 were females. The age of onset was 1.8 (1.0, 3.2) years. The rate of CAA by Z score criteria was higher than that by traditional method (21.9% (529/2 419) vs. 13.9% (336/2 419), χ 2 =1 074.94, P< 0.001). Compared to the traditional method, the Z score criteria found higher rate of CAA in male patients, patients with incomplete Kawasaki disease, and IVIG-unresponsive patients (25.2% (392/1 558) vs. 16.0% (249/1 558), (32.7% (166/507) vs. 19.5% (99/507), 30.5% (95/312) vs. 24.0% (75/312), χ 2 =694.05, 216.19, 184.37, all P< 0.001). The Z score criteria was consistent with the traditional method in diagnosing CAA (κ=0.642, P <0.001). Moreover, in the Z score criteria, the rate of CAA in males (25.2%, 392/1 558) was higher than that in females (15.9%, 137/861), higher in incomplete Kawasaki cases (32.7%, 166/507) than that in complete Kawasaki case (19.0%, 363/1 912), and higher in IVIG-unresponsive cases (30.4%, 95/312) than that in IVIG-sensitive cases (20.6%, 434/2 107), with statistically significant differences (χ 2 =27.76, 44.38, 15.43, all P <0.001). Coronary Z score of course ≤ 6 weeks was greater than that of course between>6-8 weeks and >8 weeks to 6 months (1.3 (0.7, 2.3) vs. 0.7 (0.3, 1.4), 0.7 (0.3, 1.3), Z= 20.65, 13.70, both P< 0.001). Conclusions: The rate of CAA in Kawasaki disease by Z score criteria is higher than that by traditional method. In the Z score group, most CAA occur within 6 weeks of the course of the disease, and the rate of CAA in male, incomplete Kawasaki disease, and IVIG-unresponsive is higher.

Objective: To detect the incidence and analyze the clinical significance of regions of homozygosity (ROH) through the single nucleotide polymorphism array (SNP array). Methods: The SNP array detection results of 5 116 pregnant women in the Third Affiliated Hospital of Guangzhou Medical University from January 2016 to December 2020 were retrospectively analyzed. The pregnant women with ROH (5 Mb as the threshold) were followed up to analyze the relationship between ROH and abnormal fetal phenotype. Whole exon sequencing was performed in 4 cases of consanguineous marriage to detect potential recessive causative genes in the ROH region. Results: (1) A total of 39 cases of ROH were detected, with a positive rate of 0.76% (39/5 116). Among them, 25 cases (64%, 25/39) were detected only on single chromosome, and chromosome 11 had the highest detection rate, suggesting the risk of uniparental disomy; fourteen cases (36%,14/39) were detected on multiple chromosomes, most commonly on chromosomes 11, 1, 3, 4 and 8. (2) The number of cases and detection rate of ROH detected by different prenatal diagnosis indicators were as follows: 12 cases (1.78%, 12/676) in pregnant women with abnormal non-invasive prenatal testing result, 12 cases (0.37%, 12/3 284) in pregnant women with ultrasound abnormality, 4 cases (4/4) in pregnant women with consanguineous marriage, 3 cases (0.92%, 3/326) in pregnant women with previous adverse pregnancy, 2 cases (1.15%, 2/174) in pregnant women with high risk of serology in screening, 2 cases (4.00%, 2/50) in pregnant women with abnormal fetal chromosomal karyotype, 2 cases (0.79%, 2/253) in pregnant women with advanced maternal age, 1 case (0.56%, 1/178) in pregnant women with related parental genetic factors and 1 case (0.58%, 1/171) in pregnant women with the other factors. (3) The follow-up results of 39 cases of prenatal ROH showed that there were 16 cases of term birth, 15 cases of termination of pregnancy, 2 cases of preterm births, 1 case of fetal death and 5 cases lost to follow-up. Conclusions: Chromosomal ROH phenomenon is not rare. By analyzing the detection rate of ROH in prenatal diagnosis, combined with the results of fetal phenotype and postpartum follow-up, the clinical characteristics of ROH are discussed, so as to better understand the relationship between ROH and its phenotype.

Objective: To investigate the effect of triptolide on radiosensitivity of lung cancer cells and its mechanism. Methods: The lung cancer cells H1299, A549, H157 and H838 were cultured. The strongest radio resistance cell line, H1299 was selected by cell clone formation experiment and for the subsequent experiments. 3-(4, 5-dimethyl-2-thiazolyl)-2, 5-diphenyl-2H tetrazolium bromide (MTT) was used to detect the effect of different concentrations of triptolide on the proliferation of H1299 cells. The optimal concentration of triptolide was 50nmol/L, and the optimal treatment time was 48 hours. The H1299 cells were divided into the control group, triptolide group (50 nmol/L), 4 Gy group and triptolide+ 4 Gy group. Flow cytometry was used to detect the apoptosis rate of H1299 cell. Western Blot was used to detect the proteins expression levels of p-Chk2, p-ATM, p-p53, Bcl-2, Bax and cleaved-Caspase 3. Results: The apoptotic rate and protein levels of Bax, cleaved-Caspase 3, p-Chk2, p-ATM and p-p53 in the 4 Gy group were (12.38±1.34)%, 0.42±0.04, 0.38±0.04, 0.98±0.11, 0.73±0.08, 0.95±0.09, respectively, higher than (3.26±2.43)%, 0.22±0.02, 0.23±0.03, 0.32±0.03, 0.21±0.02, 0.25±0.03 in the control group ( P <0.05). However, the protein level of Bcl-2 was (0.52±0.05), lower than (0.93±0.09) of the control group ( P <0.05). The survival fraction (0.462) and protein level of Bcl-2 (0.44±0.04) in the triptolide group were lower than those of the control group (0.702 and 0.93±0.09, P <0.05). The apoptotic rate and the protein levels of Bax and cleaved-Caspase 3 in the triptolide group were (9.27±1.08)%, 0.45±0.05, 0.41±0.04, respectively, higher than (3.26±2.43)%, 0.22±0.02, 0.23±0.03 in the control group ( P <0.05), and the sensitization ratio in the triptolide group was 1.579. The apoptosis rate, Bax and cleaved Caspase 3 protein expression levels in triptolide + 4 Gy group were (26.53±2.19)%, 0.91±0.09 and 0.79±0.08, respectively, higher than (12.38±1.34)%, 0.42±0.04 and 0.38±0.04 in 4 Gy group ( P <0.05). The expression level of Bcl-2 protein was (0.21±0.02), lower than (0.52±0.05) in 4 Gy group ( P <0.05). Conclusion: Triptolide increases the radiosensitivity of radiation-induced lung cancer cells by inhibiting DNA repair and inducing apoptosis.

Aims: This study aimed to explore potential drug targets of Streptococcus suis at the system level., Methods and Results: A homologous protein mapping method was used in the construction of a protein-protein interaction (PPI) network of S. suis, which presented 1147 non-redundant interaction pairs among 286 proteins. The parameters of PPI networks were calculated and showed scale-free network properties. In all, 41 possibly essential proteins identified from 47 highly connected proteins were selected as potential drug target candidates. Of these proteins, 30 were already regarded as drug targets in other bacterial species. Six transporters with high connections to other functional proteins were identified as probably not essential but important functional proteins. Afterward, the subnetwork centred with cell division protein FtsZ was used in confirming the PPI network through bacterial two-hybrid analysis., Conclusions: The predicted PPI network covers 13·04% of the proteome in S. suis. The selected 41 potential drug target candidates are conserved between S. suis and several model bacteria., Significance and Impact of the Study: The predictions included proteins known to be drug targets, and a verifying experiment confirmed the reliability of predicted interactions. This work is the first to present systematic computational PPI data for S. suis and provides potential drug targets, which are valuable in exploring novel anti-streptococcus drugs., (© 2020 The Society for Applied Microbiology.)

The measurement of the energy spectrum of cosmic ray helium nuclei from 70 GeV to 80 TeV using 4.5 years of data recorded by the Dark Matter Particle Explorer (DAMPE) is reported in this work. A hardening of the spectrum is observed at an energy of about 1.3 TeV, similar to previous observations. In addition, a spectral softening at about 34 TeV is revealed for the first time with large statistics and well controlled systematic uncertainties, with an overall significance of 4.3σ. The DAMPE spectral measurements of both cosmic protons and helium nuclei suggest a particle charge dependent softening energy, although with current uncertainties a dependence on the number of nucleons cannot be ruled out.

Abstract: Objective To study the changes of metabolites in serum and tissues （kidney, liver and heart） of mice died of acute tetracaine poisoning by metabolomics, to search for potential biomarkers and related metabolic pathways, and to provide new ideas for the identification of cause of death and research on toxicological mechanism of acute tetracaine poisoning. Methods Forty ICR mice were randomly divided into control group and acute tetracaine poisoning death group. The model of death from acute poisoning was established by intraperitoneal injection of tetracaine, and the metabolic profile of serum and tissues of mice was obtained by ultra-high performance liquid chromatography-electrostatic field orbitrap high resolution mass spectrometry （UPLC-Orbitrap HRMS）. Multivariate statistical principal component analysis （PCA） and orthogonal partial least square-discriminant analysis （OPLS-DA） were used, combined with t-test and fold change to identify the differential metabolites associated with death from acute tetracaine poisoning. Results Compared with the control group, the metabolic profiles of serum and tissues in the mice from acute tetracaine poisoning death group were significantly different. Eleven differential metabolites were identified in serum, including xanthine, spermine, 3-hydroxybutylamine, etc.; twenty-five differential metabolites were identified in liver, including adenylate, adenosine, citric acid, etc.; twelve differential metabolites were identified in heart, including hypoxanthine, guanine, guanosine, etc; four differential metabolites were identified in kidney, including taurochenodeoxycholic acid, 11, 12-epoxyeicosatrienoic acid, dimethylethanolamine and indole. Acute tetracaine poisoning mainly affected purine metabolism, tricarboxylic acid cycle, as well as metabolism of alanine, aspartic acid and glutamic acid. Conclusion The differential metabolites in serum and tissues of mice died of acute tetracaine poisoning are expected to be candidate biomarkers for this cause of death. The results can provide research basis for the mechanism and identification of acute tetracaine poisoning., 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 Forensic Medicine.)

The article "Preclinical pharmacokinetic study of a novel lipid-lowering agent, IMM-H007, by Z.-L. Zhang, W.-Q. Liu, X.-Z. Deng, published in Eur Rev Med Pharmacol Sci 2018; 22 (24): 8939-8950-DOI: 10.26355/eurrev&#95;201812&#95;16664-PMID: 30575938" has been withdrawn from the authors stating that "after our article was published, we received a conflict of interest from the Chinese Academy of Sciences. They declared that they developed IMM-H007 for the first time and they had already applied for the patent. We had already cited in the references that IMM-H007 came from the Chinese Academy of Sciences. But they still insisted that we had violated their rights. We had to withdraw our paper in order to avoid greater conflicts and disputes". The Publisher apologizes for any inconvenience this may cause. https://www.europeanreview.org/article/16664.

Several bioactive molecules isolated from the saliva of blood-sucking arthropods, such as mosquitoes, have been shown to exhibit potential anticoagulant function. We have previously identified a 30kDa allergen named Aegyptin-like protein (alALP), which is highly homologous to Aegyptin, from the salivary glands of female Aedes albopictus (Asian tiger mosquito). In this study, we identified the conserved functional domain of alALP by using bioinformatic tools, and expressed the His-tagged alALP recombinant protein in sf9 insect cells by generation and transfection of a baculoviral expression plasmid carrying the fulllength cDNA of alALP. We purified this recombinant protein and examined its function on the inhibition of blood coagulation. The results showed that the purified His-alALP prolonged the Activated Partial Thromboplastin Time (APTT), Prothrombin Time (PT) and Thrombin Time (TT) in vitro as well as the Bleeding Time (BT) in vivo, which suggest that alALP could be a novel anticoagulant.

The precise measurement of the spectrum of protons, the most abundant component of the cosmic radiation, is necessary to understand the source and acceleration of cosmic rays in the Milky Way. This work reports the measurement of the cosmic ray proton fluxes with kinetic energies from 40 GeV to 100 TeV, with 2 1 / 2 years of data recorded by the DArk Matter Particle Explorer (DAMPE). This is the first time that an experiment directly measures the cosmic ray protons up to ~100 TeV with high statistics. The measured spectrum confirms the spectral hardening at ~300 GeV found by previous experiments and reveals a softening at ~13.6 TeV, with the spectral index changing from ~2.60 to ~2.85. Our result suggests the existence of a new spectral feature of cosmic rays at energies lower than the so-called knee and sheds new light on the origin of Galactic cosmic rays., (Copyright © 2019 The Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science. No claim to original U.S. Government Works. Distributed under a Creative Commons Attribution NonCommercial License 4.0 (CC BY-NC).)

Objective: The aim of this study was to investigate the effect of long non-coding ribonucleic acid (lncRNA) nuclear paraspeckle assembly transcript 1 (NEAT1) on sepsis-induced brain injury in mice through nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB)., Materials and Methods: The mouse model of sepsis was established by cecal ligation and puncture induction. The relative expression levels of NEAT1 and NF-κB in brain tissues of mice in healthy group and sepsis group were determined via quantitative Reverse Transcription-Polymerase Chain Reaction (qRT-PCR) and Western blotting, respectively. Subsequently, the expression of NEAT1 was silenced by transfection of small interfering RNAs (siRNAs). Meanwhile, its effect on NF-κB expression was detected. To further explore the effect of sepsis on brain injury, the content of brain water and the expression levels of apoptosis-related proteins, including B-cell lymphoma 2 (Bcl-2) and Bcl-2-associated X protein (BAX), in mice of healthy group, sepsis group, and sepsis + si-NEAT1 group were measured. Furthermore, MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide) and terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assays were used to detect the proliferation and apoptosis of nerve cells., Results: The relative expression levels of NEAT1 and NF-κB were significantly increased in the brain tissues of septic mice (p<0.01). Si-NEAT1 transfection significantly decreased the expressions of NEAT1 and NF-κB in brain tissues of septic mice (p<0.05). The content of brain water in mice of sepsis group was evidently increased (p<0.05). However, si-NEAT1 treatment remarkably reduced this content (p<0.05). In addition, sepsis markedly decreased the activity of nerve cells (p<0.05). However, si-NEAT1 could significantly increase the activity of nerve cells in septic mice (p<0.05). Moreover, si-NEAT1 notably decreased the expression of BAX (p<0.05), whereas it increased the expression of Bcl-2 (p<0.05). The results of apoptosis detection revealed that sepsis remarkably promoted the apoptosis of mouse nerve cells (p<0.05). In addition, si-NEAT1 transfection could evidently alleviate the apoptosis of nerve cells in septic mice (p<0.05)., Conclusions: LncRNA NEAT1 promotes brain injury in septic mice by positively regulating NF-κB. However, si-NEAT1 transfection can reduce this injury.

Heavy metal pollution from industrial wastewater is an important source. A method for heavy metals determination in industrial wastewater based on laser-induced breakdown spectroscopy (LIBS) technique was studied and the on-line monitoring system that used automatic graphite enrichment and spatial plasma confinement detection was developed and field demonstrated. The limits of detection (LOD) of heavy metal elements (Cd, Cr, Cu, Ni, Pb, Zn) could reach several μg/L. In Tongling, the on-line heavy metal monitor was field demonstrated. The calibration curves of copper and zinc were built on site, and then on-line monitoring was conducted. The measurement results of this monitor were compared with ICP-OES and had a good correlation. The results showed that the heavy metal monitor could be used for on-line detection of heavy metals in wastewater and had a good reliability.

Objective: To investigate the pharmacokinetic characteristics of absorption, distribution, metabolism, and excretion in vivo after oral administration and sublingual venous injection of the small molecule IMM-H007 in hamsters. MATERIALS AND  METHODS: Pharmacokinetic characteristics, including absorption, distribution, metabolism, and excretion, were studied in vivo by LC-MS/MS after oral administration and sublingual venous injection of IMM-H007 in hamsters. Furthermore, IMM-H007 stability in artificial gastric juices, artificial intestinal juices, and Tris-HCl buffer was also analyzed., Results: There was no significant matrix or impurity interference in golden hamster whole blood as shown using MS/MS analysis to detect the existence of these substances. IMM-H007, Ml, and MP exhibited good linearity in the range of 1-500 ng/mL, 2-1000 ng/mL, and 10-5000 ng/mL, respectively. The matrix effect was 71.93-105.49%, and IMM-H007, M1, and MP were stable during the process of sample disposal. These results illustrate that the HPLC MS/MS analytic method is simple, reliable, and sensitive and exhibits high specificity and which meets the clinical pharmacokinetic requirements of IMM-H007. IMM-H007 is rapidly absorbed through the oral route in hamsters. The Cmax and AUC(0-t) of the Ml and MP metabolites in male and female hamsters were increased with increasing dosage and were proportional to the dose. In addition, T1/2 and MRT(0-t) were significantly prolonged with increasing dosage, exhibiting linear dynamic characteristics and no significant gender differences. Bioavailability in male and female golden hamsters after oral administration of IMM-H007 was calculated using the sum of Ml and MP, resulting in 6.97% and 8.95%, respectively. IMM-H007 and its metabolites were stable in Tris-HCl buffer, artificial gastric juices, and artificial intestinal juices., Conclusions: We provide an experimental basis for elucidating the material pharmacodynamics actions of IMM-H007 and predicting its potential drug interactions.

Successful spin injection into graphene makes it a competitive contender in the race to become a key material for quantum computation, or the spin-operation-based data processing and sensing. Engineering ferromagnetic metal (FM)/graphene heterojunctions is one of the most promising avenues to realise it, however, their interface magnetism remains an open question up to this day. In any proposed FM/graphene spintronic devices, the best opportunity for spin transport could only be achieved where no magnetic dead layer exists at the FM/graphene interface. Here we present a comprehensive study of the epitaxial Fe/graphene interface by means of X-ray magnetic circular dichroism (XMCD) and density functional theory (DFT) calculations. The experiment has been performed using a specially designed FM1/FM2/graphene structure that to a large extent restores the realistic case of the proposed graphene-based transistors. We have quantitatively observed a reduced but still sizable magnetic moments of the epitaxial Fe ML on graphene, which is well resembled by simulations and can be attributed to the strong hybridization between the Fe 3dz2 and the C 2pz orbitals and the sp-orbital-like behavior of the Fe 3d electrons due to the presence of graphene.

Objectives: To compare adverse effects, tolerability and efficacy of the tricyclic antidepressants (TCAs) amitriptyline and nortriptyline in management of neuropathic pain due to peripheral neuropathy (PN)., Materials & Methods: We performed a prospective open-label flexible-dosing comparison of monotherapy or adjuvant therapy using amitriptyline or nortriptyline in PN-associated neuropathic pain. Primary outcomes were quantitative adverse effects and discontinuation rates. Secondary outcomes assessed changes in pain severity, quality of life, disability, sleep efficacy, mood and anxiety, and global improvement. Assessments occurred at 3 and 6 months after initiation. Our hypothesis was that nortriptyline would have better tolerance than amitriptyline., Results: A total of 228 PN patients were enrolled approximately equally for monotherapy and adjuvant therapy. Adverse effects and discontinuation rates were similar between amitriptyline and nortriptyline interventions. Weight gain was more common with amitriptyline, while nortriptyline use was associated with greater prevalence of dry mouth. Secondary outcome measures were similar in both groups, demonstrating improvement from baseline., Conclusions: Amitriptyline and nortriptyline are equivalent for overall adverse effects and discontinuation rates. Either TCA should be equally considered for use in neuropathic pain due to PN. When used as monotherapy or as part of adjuvant therapy, either TCA can be expected to provide approximately 23-26% visual analog scale pain reduction if tolerated. Discontinuations due to inefficacy or adverse effects can be anticipated in 26-37% of patients initiated on either TCA for PN-associated neuropathic pain., (© 2013 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

The magnetic properties of a Co2FeAl/(Ga,Mn)As bilayer epitaxied on GaAs (001) are studied both experimentally and theoretically. Unlike the common antiferromagnetic interfacial interaction existing in most ferromagnet-magnetic semiconductor bilayers, a ferromagnetic interfacial interaction in the Co2FeAl/(Ga,Mn)As bilayer is observed from measurements of magnetic hysteresis and x-ray magnetic circular dichroism. The Mn ions in a 1.36 nm thick (Ga,Mn)As layer remain spin polarized up to 400 K due to the magnetic proximity effect. The minor loops of the Co2FeAl/(Ga,Mn)As bilayer shift with a small ferromagnetic interaction field of +24 Oe and -23 Oe at 15 K. The observed ferromagnetic interfacial coupling is supported by ab initio density functional calculations. These findings may provide a viable pathway for designing room-temperature semiconductor spintronic devices through magnetic proximity effect.

Hydrogen sulfide (H(2)S), an endogenous "gasotransmitter", exists in the central nervous system. However, the central cardiovascular effects of endogenous H(2)S are not fully determined. The present study was designed to investigate the central cardiovascular effects and its possible mechanism in anesthetized rats. Intracerebroventricular (icv) injection of NaHS (0.17~17 microg) produced a significant and dose-dependent decrease in blood pressure (BP) and heart rate (HR) (P < 0.05) compared to control. The higher dose of NaHS (17 microg, n = 6) decreased BP and HR quickly of rats and 2 of them died of respiratory paralyse. Icv injection of the cystathionine beta-synthetase (CBS) activator s-adenosyl-L-methionine (SAM, 26 microg) also produced a significant hypotension and bradycardia, which were similar to the results of icv injection of NaHS. Furthermore, the hypotension and bradycardia induced by icv NaHS were effectively attenuated by pretreatment with the K(ATP) channel blocker glibenclamide but not with the CBS inhibitor hydroxylamine. The present study suggests that icv injection of NaHS produces hypotension and bradycardia, which is dependent on the K(ATP) channel activation.

The frequent disease outbreaks caused by avian influenza virus not only affect the poultry industry but also pose a threat to human safety. To address the problem, RNA interference (RNAi) has recently been widely used as a potential antiviral approach. Transgenesis in combination with RNAi to specifically inhibit avian enza virus gene expression has been proposed to make chickens resistant to the infection. For the transgenic breeding, screening in vitro efficient siRNAs as the candidate genes is one of the most important tasks. Here, we combined an online search tool and a series of bioinformatics programs with a set of rules for designing siRNAs targeted towards different mRNA regions of H5N1 avian influenza virus. Five rational siRNAs were chosen by this method, five U6 promoter-driven shRNA expression plasmids containing the siRNA genes were constructed and used for producing stably transfected MDCK cells. The data obtained by virus titration, IFA, PI-stained flow cytometry, real-time quantitative RT-PCR, and DAS-ELISA analyses showed that all five stably transfected cell lines we re resistant to virusreplication when exposed to 100 CCID50 of avian influenza virus H5N1. Finally, most effective plasmids (pSi-604i and pSi-1597i) as the candidates for making the transgenic chickens were chosen. These findings provide baseline information on use of RNAi technique for breeding transgenic chickens resistant to avian influenza virus.

The frequent disease outbreaks caused by avian influenza virus (AIV) not only affect the poultry industry but also pose a threat to human safety. To address the problem, RNA interference (RNAi) has recently been widely used as a potential antiviral approach. Transgenesis, in combination with RNAi to specifically inhibit AIV gene expression, has been proposed to make chickens resistant to avian influenza. For the transgenic breeding, screening the efficient siRNAs in vitro as the candidate genes is one of the most important tasks. Here, we combined an online search tool and a series of bioinformatics programs with a set of rules for designing the siRNAs targeting different mRNA regions of AIV H5N1 subtype. By this method we chose five rational siRNAs, constructed five U6 promoter-driven shRNA expression plasmids contained the siRNA genes, and used these to produce stably transfected Madin-Darby canine kidney (MDCK) cells. Data from virus titration, IFA, PUI-stained flow cytometry, real-time quantitative RT-PCR and DAS-ELISA analyses showed that all five stably transfected cell lines were effectively resistant to viral replication when exposed to 100 CCID50 of AIV, and we finally chose the most effective plasmids (pSi-604i and pSi-1597i) as the candidates for making the transgenic chickens. These findings provide baseline information for breeding transgenic chickens resistant to AIV in combination with RNAi., (© Pleiades Publishing, Ltd. 2010.)

We identify an essential role for the RGD (Arg-Gly-Asp tripeptide) moiety in vivo during adult peripheral neuron regenerative growth. Beyond a peripheral nerve transection there were rises in the fibronectin extracellular matrix, and striking rises in the mRNA and protein expression of integrin subunits sensitive to RGD/fibronectin signalling. Neuron perikarya, axons and Schwann cells all expressed RGD/fibronectin sensitive integrins after injury. To evaluate the significance of RGD/fibronectin-integrin interactions, we infused solutions of a pentapeptide including the RGD motif (sRGD) serially and directly within the milieu of early axon growth across rat sciatic transection injuries. While low dose infusions of sRGD facilitated early axon ingrowth, we encountered inhibition of ingrowth and bridge formation with higher doses of sRGD indicating competitive disruption of RGD/fibronectin-integrin signalling. Fibronectin RGD moieties serve a critical and important role during peripheral axon outgrowth.

This study investigated the nucleotide sequences and tissue expression levels of genes relating to the ovulation rate in Yunling black goats, a famous Yunnan province, China, local breed with low fecundity. Five genes, FSHB, FSHR, BMP15, BMPR1B, and ESR2, were investigated; the complete cDNA sequences of these genes were 390, 2088, 1185, 1509, and 1585 bp, respectively, and compared with Boer goats (a more fecund breed), the sequence identities were 99%, 99%, 99%, 100%, and 99%, respectively. There were two base differences in FSHB and BMP15, four in FSHR, and three in ESR2. There were fewer follicles and oocytes in Yunling black goats than in Boer goats. Expression levels of FSHB, FSHR, and BMP15 genes in Yunling black goats were lower, and expression levels of BMPR1B and ESR2 genes were higher. Serum FSH content was lower in Yunling black goats, but serum estrogen content was higher. Protein expression levels of FSHR, BMP15, BMPR1B, and ESR2 in ovary and pituitary correlated positively with gene mRNA expression levels. In Yunling black goats, the mRNA expression levels of FSHB, FSHR, and BMP15 positively correlated with litter size, but those of BMPR1B and ESR2 correlated negatively. Together, base changes and variations in mRNA and protein expression levels of genes relating to the ovulation rate result in low fecundity in the Yunling black goat. Reduced BMP15 and FSHR levels may be related to the observed fewer oocytes and, consequently, fewer follicles.

Amyotrophic lateral sclerosis (ALS) is a fatal neuromuscular disorder in which motor neurons may be targeted by oxidative and nitrergic stress without sufficient compensation by intrinsic support mechanisms. In this work, we addressed two key tenets of this hypothesis for the pathogenesis of ALS. Using superoxide dismutase (SOD) 1G93A mice, we studied the impact of reduction of nitrergic stress within the CNS with the use of a broad spectrum nitric oxide synthase (NOS) inhibitor, NG-nitro-l-arginine methyl ester. A separate cohort of SOD1G93A mice received direct insulin neurotrophic support, ligating receptors expressed upon motor neurons, to attempt protection against neuronal and functional motor dropout. For direct access, we used a novel form of intranasal delivery that provides peak concentration levels in the CNS within 1 h of delivery without systemic side effects at doses which previously rescued retrograde loss of motor axons after axotomy. To identify even minor impacts of these interventions on the outcome, we utilized an intensive program of serial behavioral and electrophysiological testing weekly, combined with endpoint quantitative morphometry and molecular analysis. This intensive evaluation enhanced our knowledge of the time course in SOD1G93A mice and impact of the SOD1G93A mutation upon motor neurons and their function. Neither intervention had even minimal impact upon slowing progression of disease in SOD1G93A mice. Our data argue against significant roles for nitrergic stress in promoting motor neuron loss and the importance of alternative neurotrophic support mechanisms that might support motor neurons and prevent disease progression in SOD1G93A mice.

The regeneration of adult peripheral neurons after transection is slow, incomplete and encumbered by severe barriers to proper regrowth. The role of RHOA GTPase has not been examined in this context. We examined the expression, activity and functional role of RHOA GTPase and its ROK effector, inhibitors of regeneration, during peripheral axon outgrowth. We used qRT-PCR, quantitative immunohistochemistry, and assays of RHOA activation to examine expression in sensory neurons of rats with sciatic transection injuries. In vitro, we exposed dissociated adult sensory neurons, not grown on inhibitory substrates, to a RHOA-ROK inhibitor HA-1077 and measured neurite initiation and outgrowth. In vivo, we exposed early regenerating axons and Schwann cells directly to HA-1077 in a conduit connecting the proximal and distal stumps of transected sciatic nerves. Intact adult dorsal root ganglia sensory neurons expressed RHOA and ROK 1 mRNAs and protein and there were rises in RHOA after injury. Activated GTP-bound RHOA, undetectable in intact ganglia, was dramatically upregulated in both neurons and axons after injury. Adult rat sensory neurons in vitro demonstrated a dose-related increase in the initiation of neurite outgrowth, and in the proportion with long neurites when they were exposed to a ROK antagonist. Regenerative bridges that were directly exposed to the ROK inhibitor had a dose-related rise in the extent and distance of in vivo axon and partnered Schwann cell regrowth within them. RHOA activation and signaling are features of adult peripheral axon regeneration within its own milieu, independent of myelin. Inhibition of its activation may benefit peripheral axon lesions.

Erythropoietin (EPO) and its receptor (EPO-R), mediate neuroprotection from axonopathy and apoptosis in the peripheral nervous system (PNS). We examined the impact and potential mechanisms of local EPO signaling on regenerating PNS axons in vivo and in vitro. As a consequence of injury, peripheral nerve axons and DRG neurons have a marked increase in the expression of EPO and EPO-R. Local delivery of EPO via conduit over 2 weeks to rat sciatic nerve following crush injury increased the density and maturity of regenerating myelinated axons growing distally from the crush site. In addition, EPO also rescued retrograde degeneration and atrophy of axons. EPO substantially increased the density and intensity of calcitonin gene-related peptide (CGRP) expression within outgrowing axons. Behavioral improvements in sensorimotor function also occurred in rats exposed to near nerve EPO delivery. EPO delivery led to decreased nuclear factor kappaB (NFkB) activation but increased phosphorylation of Akt and STAT3 within nerve and dorsal root ganglia neurons indicating rescue from an injury phenotype. Spinal cord explant studies also demonstrated a similar dose-dependent effect of EPO upon motor axonal outgrowth. Local EPO signaling enhances regenerating peripheral nervous system axons in addition to its known neuroprotection. Exogenous EPO may have a therapeutic role in a large number of peripheral nerve diseases through its impact on regeneration.

A charge amplifier or a voltage amplifier can be used as a signal conditioning circuit between a piezoelectric element and a meter or a data acquisition board. The outputs of the piezoelectric sensor are in an open-circuit state and a short-circuit state if a voltage amplifier and a charge amplifier are used, respectively. When the electrodes are in different states, the piezoelectric element has rather different stiffness and thus different sensor resonant frequency. This phenomenon is theoretically analyzed in detail and validated by a carefully designed experiment. The results indicate that a much wider range of working frequency is achieved when a voltage amplifier is used.

Growth factor receptor-bound protein 2 (Grb2) is an adapter protein involved in the Ras-dependent signaling pathway that plays an important role in human cancers initiated by oncogenic receptors. Grb2 is constituted by one Src homology 2 domain surrounded by two SH3 domains, and the inhibition of the interactions produced by these domains could provide an antitumor approach. In evaluating chemical libraries, to search for potential Grb2 inhibitors, it was necessary to elaborate a rapid test for their screening. We have developed, first, a batch method based on the use of an affinity column bearing a Grb2-SH3 peptide ligand to isolate highly purified Grb2. We subsequently describe a very rapid 96-well screening of inhibitors based on a simple competition between purified Grb2 and a peroxidase-coupled proline-rich peptide.

This paper describes the design of the highest affinity ligands for Grb2 SH3 domains reported so far. These compounds were designed by combining N-alkyl amino acid incorporation in a proline-rich sequence with subsequent dimerization of the peptoid sequence based on structural data and molecular modeling. Optimization of the linker size is discussed, and the N-alkyl amino acid incorporation into both monomeric halves is reported. Because the affinity for Grb2 of the optimized compounds was too high to be measured using the fluorescent modifications that they induce on the Grb2 emission spectrum, a competition assay was developed. In this test, Grb2 is pulled down from a cellular extract by the initial VPPPVPPRRR peptide bound to Sepharose beads. In the presence of competitors, the test quantifies the amount of Grb2 displaced from the beads. It has enabled us to determine a K(i) value in the 10(-10) M range for the highest affinity Grb2 peptoid analogue dimer.

Recently a strong positive association between schizophrenia and Notch4 has been reported. Both individual markers and haplotypes showed association with the disease, with five markers (three microsatellites and two SNPs) being tested. In order to test this finding we genotyped these markers in the Han Chinese population using a sample of 544 cases and 621 controls as well as >300 trios. Analysis of allele, genotype and haplotype frequencies in both samples showed no association between the markers and the disease. Our results would indicate that a significant role for the Notch4 gene in schizophrenia can be ruled out in the Han Chinese. However, similar studies are necessary in the Caucasian population as linkage disequilibrium arrangements and founder effects may differ between these two populations.

In the vitamin K-dependent protein family, only protein S (PS) contains a thrombin-sensitive region (TSR), located between the domain containing the gamma-carboxyglutamic acid and the first epidermal growth factor-like domain. To better define the role of TSR in the PS molecule, we expressed a recombinant human PS (rHPS) and its analogue lacking TSR (rTSR-less), and prepared factor Xa- and thrombin-cleaved rHPS. A peptide reproducing TSR (TSR-peptide) was also synthesized in an attempt to obtain direct evidence of the domain involvement in PS anticoagulant activity. In a coagulation assay, both rTSR-less and factor Xa-cleaved PS were devoid of activated protein C cofactor activity. The TSR-peptide did not inhibit rHPS activity, showing that TSR must be embedded in the native protein to promote interaction with activated protein C. The binding of rHPS to activated platelets and to phospholipid vesicles was not modified after factor Xa- or thrombin-mediated TSR cleavage, whereas the binding of rTSR-less was markedly reduced. This suggested a role for TSR in conferring to PS a strong affinity for phospholipid membranes. TSR-peptide did not directly bind to activated platelets or compete with rHPS for phospholipid binding. The results of the present study show that TSR may not interact directly with membranes, but probably constrains the gamma-carboxyglutamic acid-rich domain in a conformation allowing optimal interaction with phospholipids.

Objective: To explore the activity of heme oxygenase and immunolocate the enzyme in the adult worms of Schistosoma japonicum., Methods: Microsomal protein was isolated from the homogenate of adult S. japonicum, heme degradation and effect of different pH conditions and buffers on degrading reaction were investigated by incubating microsomal protein with hemin. The slices of whole worm and cells of S. japonicum were prepared, distribution of HO in schistosome was studied by immunofluorescent and alkaline phosphatase(AP)-immunocytochemical assays., Results: Microsomal protein of adult worms can degrade the heme in vitro, the activity being 56.7 nmol bilirubin/(mg.min). The optimal pH was 8.7. Immunofluorescent and AP-immunocytochemical assays revealed that the HO distributed dispersively in the worm, and located in cytoplasm., Conclusion: The presence of HO was firstly proved in S. japonicum.

Anarchic cell proliferation, observed in some leukemia and in breast and ovarian cancers, has been related to dysfunctioning of cytoplasmic or receptor tyrosine kinase activities coupled to p21 Ras. The growth factor receptor-bound protein 2 (Grb2) adaptor when complexed with Sos (Son of sevenless), the exchange factor of Ras, conveys the signal induced by tyrosine kinase-activated receptor to Ras by recruiting Sos to the membrane, allowing activation of Ras. This review shows how it is possible to stop the Ras-deregulated signaling pathway to obtain potential antitumor agents. Grb2 protein is comprised of one SH2 surrounded by two SH3 domains and interacts by means of its Src homology (SH2) domain with phosphotyrosine residues of target proteins such as the epidermal growth factor (EGF) receptor or the Shc adaptor. By means of its SH3 domains, Grb2 recognizes proline-rich sequences of Sos, leading to Ras activation. Inhibitors of SH2 and SH3 domains were designed with the aim of interrupting Grb2 recognition. On the one hand, using structural data and molecular modeling, peptide dimers or "peptidimers", made up of two proline-rich sequences from Sos linked by an optimized spacer, were developed. On the other, using the structure of the Grb2 SH2 domain complexed with a phosphotyrosine (pTyr)-containing peptide and molecular modeling studies, a series of N-protected tripeptides containing two phosphotyrosine or mimetic residues, with one pTyr sterically constrained, were devised. These compounds show very high affinities for Grb2 in vitro. They have been targeted into cells showing selective antiproliferative activity on tumor cells. These results suggest that inhibiting SH2 or SH3 domains of signaling proteins might provide antitumor agents.

Phosphopeptide prodrugs bearing two S-acyl-2-thioethyl (SATE) biolabile phosphate protections were developed. They are capable to inhibit the Shc/Grb2 interaction and MAP kinases (ERK1 and ERK2) phosphorylation in cellular assay. The S-acetyl-2-thioethyl (MeSATE) analogue showed an IC50 of 1 microM in the inhibition of the colony formation of tumor cell line NIH3T3/HER2.

A series of small peptides with the sequence mAZ-pTyr-Xaa-Asn-NH(2), where Xaa denotes alpha-methylphosphotyrosine or its carboxylic mimetics, were synthesized as inhibitors of the Grb2 SH2 domain. Peptide 3 with (alpha-Me)pTyr as Xaa has the highest affinity for Grb2 (K(d) = 3 +/- 1 nM) and exhibits to date the best inhibitory activity (IC(50) = 11 +/- 1 nM) to displace PSpYVNVQN-Grb2 interaction in an ELISA test. The lower affinities of peptides with (alpha-Me)Tyr, (alpha-Me)Phe(4-CO(2)H), or (alpha-Me)Phe(4-CH(2)CO(2)H) as Xaa demonstrate the importance of a double charged phosphate group at the pY+1 position. Molecular modeling showed additional hydrogen bond interactions provided by the (alpha-Me)pTyr residue with the Grb2 SH2 domain. These results thus show that the effect of hydrophobic pY+1 residues, initially put forth to increased the binding affinities, can be further enhanced by a (-Me)pTyr residue which has both hydrophobic and hydrophilic properties.

With the aim of interrupting the growth factor-stimulated Ras signaling pathway at the level of the Grb2-Sos interaction, a peptidimer, made of two identical proline-rich sequences from Sos linked by a lysine spacer, was designed using structural data from Grb2 and a proline-rich peptide complexed with its SH3 domains. The peptidimer affinity for Grb2 is 40 nM whereas that of the monomer is 16 microM, supporting the dual recognition of both Grb2 SH3 domains by the dimer. At 50 nM, the peptidimer blocks selectively Grb2-Sos complexation in ER 22 (CCL 39 fibroblasts overexpressing epidermal growth factor receptor) cellular extracts. The peptidimer specifically recognizes Grb2 and does not interact with PI3K or Nck, two SH3 domain-containing adaptors. The peptidimer was modified to enter cells by coupling to a fragment of Antennapedia homeodomain. At 10 microM, the conjugate inhibits the Grb2-Sos interaction (100%) and MAP kinase (ERK1 and ERK2) phosphorylation (60%) without modifying cellular growth of ER 22 cells. At the same concentration, the conjugate also inhibits both MAP kinase activation induced by nerve growth factor or epidermal growth factor in PC12 cells, and differentiation triggered by nerve growth factor. Finally, when tested for its antiproliferative activity, the conjugate was an efficient inhibitor of the colony formation of transformed NIH3T3/HER2 cells grown in soft agar, with an IC50 of around 1 microM. Thus, the designed peptidimers appear to be interesting leads to investigate signaling and intracellular processes and for designing selective inhibitors of tumorigenic Ras-dependent processes.

The effect of merocyanine 540 (Mc 540) mediated photoirradiation on both neoplastic and normal hemopoietic progenitor cells was studied. Bone marrow (BM) cells from children with acute lymphoblastic leukemia (ALL), acute myeloid leukemia (AML) at initial diagnosis, ALL in remission, neuroblastoma and normal children as well as cells of Reh-6 and HL-60 cell lines were incubated with Mc 540 in the presence of human albumin (HA) and exposed to different argon laser 514 nm doses. Cell survival was estimated using Trypan Blue supravital stain following a 24-h incubation and leukemic cell lines were studied in continuous cell cultures of 4 weeks duration. Our results showed that HA protects normal BM cells from Mc 540 mediated phototoxicity. A 99.9999% inhibition of Reh-6 and HL-60 was noted at irradiation doses where the corresponding mean survival of normal BM cells was 77.4 +/- 12 and 70.3 +/- 10%, respectively. BM leukemic cells from children with ALL and AML were also very sensitive to Mc 540 photoirradiation in contrast to neuroblastoma cells where only a three-fold reduction was observed. Finally, the survival of normal BM progenitors was 38% for colony forming unit erythroid CFU-E, 37% for burst forming unit erythroid BFU-E, 55% for CFU-GM and 29% for CFU-GEMM. In conclusion it seems that Mc 540 mediated photoirradiation in neoplastic cells exerts selective cytotoxicity and can be used in ex vivo purging of malignant cells in BM.

A laser-induced fluorescence-guided biopsy system has been developed for the screening and early detection of malignancies in the female inner/outer genital tract. Fluorescence spectra were recorded during exposure of normal and malignant tissue to He-Cd laser (442 nm) radiation. A characteristic increase in the fluorescence signal at 600 nm for malignant tissue was observed after treatment of the samples with hypocrellin (HA). This, combined with the spectral distribution of tissue natural fluorescence, allowed for the development of simple algorithms, based on the intensity difference. A subsequent index of discrimination between normal and various malignant tissues has been calculated. These results suggest that monitoring of this index during colposcopy could enhance selective detection of the malignant tissue, reducing the risk of leaving pathologic tissue untreated during standard exploratory surgical procedures.

We previously showed that the oxidation of branched-chain amino acids is increased in rats treated with clofibrate [Paul and Adibi (1980) J. Clin. Invest. 65, 1285-1293]. Two subsequent studies have reported contradictory results regarding the effect of clofibrate treatment on gene expression of branched-chain keto acid dehydrogenase (BCKDH) in rat liver. Furthermore, there has been no previous study of the effect of clofibrate treatment on gene expression of BCKDH kinase, which regulates the activity of BCKDH by phosphorylation. The purpose of the present study was to investigate the above issues. Clofibrate treatment for 2 weeks resulted in (a) a 3-fold increase in the flux through BCKDH in mitochondria isolated from rat liver, and (b) a modest but significant increase in the activity of BCKDH. However, clofibrate treatment had no significant effect on the mass of E1 alpha, E1 beta, and E2 subunits of BCKDH or the abundance of mRNAs encoding these subunits. On the other hand, clofibrate treatment significantly reduced the activity, the protein mass and the mRNA levels of BCKDH kinase in the liver. In contrast to the results obtained in liver, clofibrate treatment had no significant effect on any of these parameters of BCKDH kinase in the skeletal muscle. In conclusion, our results show that clofibrate treatment increases the activity of BCKDH in the liver and the mechanism of this effect is the inhibition of gene expression of the BCKDH kinase.

Early-arriving photons of 100-fs laser pulses transmitted through highly scattering media have been detected by a streak camera. Because of their partial spatial coherence, they are affected by diffraction from small hidden discontinuities. The experimental data of the patterns are analyzed with Fresnel diffraction theory and then corrected accordingly. Submillimeter hidden objects were scanned and imaged. Diffraction correction resulted in a significantly improved contrast in the hidden object's image.

The present study demonstrates that dexamethasone and 8-(4-chlorophenylthio)adenosine 3',5'-monophosphate (CPT-cAMP), a cAMP analog, increase the substrate flux through branched-chain keto acid dehydrogenase (BCKDH) in primary rat hepatocytes cultured in defined medium. Maximum response (2.7-fold increase in flux) was observed when hepatocytes were cultured with 1 microM dexamethasone plus 50 microM CPT-cAMP for 24 h. This increase in the flux rate was accompanied by significant increases in both the basal and total activities of BCKDH (2.2- and 2.0-fold, respectively), without any significant change in the activity state of this enzyme. The increase in BCKDH activity was the result of increased protein mass of E1 alpha (3.2-fold), E1 beta (2.9-fold), and E2 (1.6-fold) subunits of BCKDH, indicating that E2 is the limiting subunit for the expression of BCKDH. The relative abundance of mRNAs encoding the E1 alpha, E1 beta, and E2 subunits of BCKDH increased by 7.4-, 21.7-, and 4.8-fold, respectively. We conclude that increased flux through BCKDH in hepatocytes cultured with dexamethasone and CPT-cAMP is due to increased expression of BCKDH subunit genes. However, nonstoichiometric expression of individual subunits and the corresponding mRNAs suggests regulation of BCKDH also at translational and post-translational steps.

A series of indicators, including serum lysozyme activity, G-banding chromosome aberration (G-banding CA) analysis, sister chromatid exchanges (SCEs), chromosome aberration (CA), T-lymphocyte transformation rate (TcTR), gamma-GT, GPT and AKP, were employed in the present survey among occupationally vinyl chloride (VC) exposed workers and inhabitants living in VC polluted area in a polyvinyl chloride (PVC) factory. The results showed that the serum lysozyme (S-LZM) activities in Group 3 (adult inhabitants exposed to 0.20 mg/m3 VC for at least 8 years), Group 2 (workers exposed to 4.1 mg/m3 for at least 7 years occupationally), Group 1 (workers exposed to 25.7 mg/m3 for at least 2 years) were significantly higher than control. G-banding CA analysis showed that the total chromosome breakage rates in both Groups 1 and 2 were higher, but no difference existed between Group 3 and control. Only Group 1 was observed having higher SCEs, CA level and lower TcTR than control. AKP levels in Groups 1 and 2 were higher than control, but no gamm-GT and GPT differences were found among groups. The study also suggests that G-banding CA analysis is more sensitive than CA and SCEs.

In order to develop a reliable laser-induced-fluorescence-guided angioplasty system, fluorescence spectra were recorded during exposure of normal and atherosclerotic cadaveric aortic tissue to He-Cd laser radiation (442 nm). A characteristic increase in the fluorescence signal at 600 nm for atheromatous tissue was observed after treatment of the samples with hypocrellin (HA). This, combined with the spectral distribution of tissue natural fluorescence, allowed the development of simple algorithms, based on the intensity difference and the full width at half-maximum (FWHM), and a subsequent index of discrimination between normal and various atheromatous tissues. Our results suggest that monitoring of this index through the catheter could enhance selective ablation, reducing the risk of normal vessel perforation.

In searching for effective anticonvulsant agent fourteen 6-(substituted-phenyl)-4,5-dihydro-3 (2H) pyridazinones and fifteen 6-(substitutedphenyl)-3 (2H) pyridazinones have been synthesized and screened in mice for their ability to antagonize maximal electroshock seizure (MES). The ED50 values showed that 6-(2',4'-dichlorophenyl)-3 (2H) pyridazinone was the most potent anticonvulsant in these compounds. The structure--activity relationships in these two series were examined. We came to the conclusion that the higher is the hydrophobic parameter pi of the substituent on phenyl ring, the more potent anticonvulsant is the compound. And also, only the compounds with an electronwithdrawing substituent on the phenyl ring exhibited appreciable anticonvulsant activity.

In recent years considerable emphasis has been placed on the hypothesis that enhancement of GABA transmission could be beneficial in some types of epilepsy. The alpha-(aryl)-4-morpholineacetonitrile obtained by the interaction of aryl aldehydes, morpholine and potassium cyanide, have been used to synthesize 3-(aroyl)-propionic acids and esters by 1,4-additions to acrylonitrile or acrylic ester. 3-(Aroyl) propionic acids reacting with hydrazine can yield 6-aryl-4,5-dihydro-3(2H) pyridazinones which are dehydrogenated by bromine (via bromination dehydrobromination) to give 6-aryl-3(2H) pyridazinones. The latter compounds were converted into 3-(N-GABA)-6-(substitutedphenyl) pyridazines and 3-(N-butyryllactamyl)-6-(substitutedphenyl) pyridazines by the chlorination (by means of phosphorus oxychloride) and then reaction with GABA. By this method seventeen 3-GABA derivatives of 6-(substituted-phenyl)pyridazines were synthesized. The anticonvulsant activity (MES) of these compounds were also tested. 3-(N-GABA)-6-(2',4'-dichloro)phenylpyridazine; potent anticonvulsant (ED50 = 21.05 mg/kg).

By injecting Naloxone, the T-lymphocyte transformation was reduced markedly. When the Naloxone plus electro-acupuncture were used together, it was not reduced but was still lower than it was when only electro-acupuncture was used. This explained why Naloxone could reverse the function by which electro-acupuncture promoted the T-lymphocyte transformation, and the endogenous opioid in the brain promoted the immune reaction. Through promoting the release of the endogenous opioid in the brain, electro-acupuncture raised the T-lymphocyte transformation function. Using electro-acupuncture when naloxone antagonized morphine, the T-lymphocyte transformation function was still promoted markedly. The mechanism could be that electro-acupuncture excited the function of the endogenous opioid system in the brain. In acupuncture immune reaction, the relationship between the endogenous opioid and catecholaminergic systems was explored.