Objective To evaluate the effect of continuous adenosine infusion on mesentery microcirculation during cardiopulmonary resuscitation in rats with asphyxia. Methods Rat model of asphyxia was established in 22 healthy Wistar rats. The animals were then randomly divided into normal saline group (group A, n=10), epinephrine group (group B, n=6) and epinephrine plus adenosine group (group C, n=6). After a 3-min asphyxia without intervention, cardiopulmonary resuscitation (CPR) was started. The cardiac compression was carried out with an electric cardio-pulmonary resuscitation machine (200 times/min). Respiration was restored and maintained with a ventilator with tidal volume of 4ml, breathing rate 50times/min and FiO2 100%. After a 4-min CPR, rats in group A and group B were given normal saline and epinephrine (bolus of 90?g/kg) respectively, and in group C epinephrine (bolus of 90?g/kg) plus 70?g/kg adenosine were given. Electric defibrillation would be initiated if there was ventricular fibrillation. The reperfusion rate of mesentery arterioles and venules, diameter of blood vessels and relative blood velocity were observed. Results The reperfusion rate of mesentery arterioles and venules was significantly higher in group C than in group B (P

This study was aimed to clone human intercellular adhesion molecule-1 (ICAM-1) gene, to transfect the constructed eukaryotic expression vector ICAM-1-GFP into CHO cells, as well as to detect ICAM-1-GFP expression in CHO cells binding with Molt-4 cells. ICAM-1 cDNA gene was amplified by RT-PCR and inserted in PMD(18)-T vector. Then ICAM-1 cDNA from pMD18-ICAM-1 vector was subcloned into eukaryotic expression vector pEGFP-C1 to construct recombinant ICAM-1-pEGFP-C1 vector. Restriction analysis and DNA sequencing were used to confirm the recombinant vector. After stable transfection of CHO-K1 cells with the recombinant vector, the expression and subcellular localization of ICAM-1-GFP were detected by RT-PCR, flow cytometry and fluorescence microscopy. The function of ICAM-1-GFP fusion protein was assessed by the binding of ICAM-1-GFP/CHO cells to Molt-4 cells. The results showed that 1622 bp full-length ICAM-1 cDNA obtained and was successfully ligated with pMD(18)-T-vector, subcloned to construct recombinant ICAM-1-pEGFP-C1 vector. Restriction analysis and DNA sequencing indicated that recombinant ICAM-1-GFP was successfully constructed and ICAM-1-GFP was expressed stably in CHO cells. ICAM-1-GFP expression was only observed in the cytoplasm of ICAM-1-GFP/CHO cells by fluorescence microscopy. The ICAM-1-GFP/CHO cells were bound to PMA-treated Molt-4 cells. The expression of MEM-148 was very weak in PMA-treated Molt-4 cells. It is concluded that the ICAM-1-GFP eukaryotic expression vector has been constructed successfully and expresses stably in CHO cells. PMA can increase the binding of Molt-4 cells to ICAM-1-GFP/CHO cells by inducing specialized form of ICAM-1 clustering.
Animals ; CHO Cells ; Cricetinae ; Cricetulus ; DNA, Complementary ; genetics ; Genetic Vectors ; Green Fluorescent Proteins ; genetics ; Humans ; Intercellular Adhesion Molecule-1 ; genetics ; Recombinant Proteins ; genetics ; Transfection

The 3-hydroxy-3-methylglutaryl-CoA reductase (HMGR) catalyzes the conversion of HMG-CoA to mevalonate in mavalonic acid pathway, which is the first committed step for isoprenoid biosynthesis in plants. However, it still remains unclear whether HGMR gene plays a role in the isoprenoid biosynthesis in Dendrobium officinale, an endangered epiphytic orchid species. In the present study, a HMGR encoding gene, designed as DoHMGR1 (GenBank accession JX272632), was identified from D. officinale using the reverse transcription polymerase chain reaction (RT-PCR) and rapid amplification of cDNA ends (RACE) methods, for the first time. The full length cDNA of DoHMGR1 was 2 071 bp in length and encoded a 562-aa protein with a molecular weight of 59.73 kD and an isoelectric point (pI) of 6.18. The deduced DoHMGR1 protein, like other HMGR proteins, constituted four conserved domains (63-561, 147-551, 268-383 and 124-541) and two transmembrane motifs (42-64 and 85-107). Multiple sequence alignment and phylogenetic analyses demonstrated that DoHMGR1 had high identity (67%-89%) to a number of HMGR genes from various plants and was closely related to Vanda hybrid cultivar, rice and maize monocots. Real time quantitative PCR (qPCR) analysis revealed that DoHMGR1 was expressed in the three included organs. The transcripts were the most abundant in the roots with 2.13 fold over that in the leaves, followed by that in the stems with 1.98 fold. Molecular characterization of DoHMGR1 will be useful for further functional elucidation of the gene involving in isoprenoid biosynthesis pathway in D. officinale.
Base Sequence ; Cloning, Molecular ; DNA, Complementary ; genetics ; Dendrobium ; enzymology ; genetics ; Gene Expression Regulation, Plant ; Hydroxymethylglutaryl CoA Reductases ; genetics ; metabolism ; Molecular Weight ; Phylogeny ; Plant Leaves ; enzymology ; genetics ; Plant Roots ; enzymology ; genetics ; Plant Stems ; enzymology ; genetics ; Plants, Medicinal ; enzymology ; genetics ; Sequence Alignment ; Sequence Homology, Amino Acid

Adult ; Carcinoid Tumor ; pathology ; Ear Neoplasms ; pathology ; Ear, Middle ; pathology ; Female ; Humans

A collection of neurons in the upper lumbar spinal cord (lumbar segments 3 and 4) of male rats project to the lower lumbar spinal cord (lumbar segments 5 and 6) and release a gastrin-releasing peptide (GRP) to the somatic and autonomic regions, which are known to regulate male sexual reflexes. The GRP plays some special functions when bound to the specific GRP receptor (GRPR). The spinal GRP system is regulated by androgens. Accumulating evidence shows that GRP plays an important role in rat penile erection and ejaculation, and pharmacological stimulation of GRPRs with a specific agonist can restore penile reflexes and ejaculation in castrated male rats. Therefore, the GRP system appears to be a potential therapeutic target for the treatment of erectile dysfunction or ejaculatory dysfunction. The present paper briefly reviews the recent studies on the role of the spinal GRP system in regulating the sexual function of males.
Androgens ; metabolism ; Animals ; Ejaculation ; physiology ; Gastrin-Releasing Peptide ; metabolism ; physiology ; Male ; Penile Erection ; physiology ; Rats ; Spinal Cord ; metabolism

Objective To evaluate the role of standard CPR training in Beijing Olympics medical volunteers by compared with the no-traimng medical personnel.Method Performance qualities of CPR was evaluated in the 44 medical vohmteers,who worked in a Beijing Olympic Venue.They received the training On standard CPR for 6 monthes,and served as training group.The performance of another 72 emergency medical workers from first-class hospitals without the training on stand CPR within 1 year(control group) Was compared with training group.Phillips QCPR3535 monitor was used to measure the compression frequency,depth and chest re-expand between the compression and the operation time via the sternal chest compression pad fitted with an accelerometer.Personal practical results of 5 circles of CPR operation were recorded in a table and the numbers ofpractical. Compression in one minute were counted.Data Were analyzed with chi-quare test and t-test.The parameters ofthe influence fac.tor(gender,age,hand placement,hand skill,compression posture and standard training)were brought into logistic regression analysis.Results Compared with the control group,the qualification rate of general CPR performance in the training group was much higher(86.4%vs.31.9%),and the compression cpmlification rates of the chestcompression,depth and chest re-palld betweenthe compression wcfe higher(88.6%,93.2%,95.5% vs.40.3%,43.0%,86.1%,P<0.01),the duration of ventilation in each operationstion was shorter[(6.38±1.3)vs.(7.57±1.6),P0.05],pratieal compressionn flberin oneminutewas not different[(77.2±3.5)vs.(77.8±12.2),P>0.05)],the ratios of the duration of compression to that of ventilation was higher(2.6:1 vs.2.1:1).Logistic analysis showed that hand skill,compression posture and standard CPR trainings were high correlated with the CPR performance qualities(P<0.05).Conclusions The performance quahty of training group is much higher thanthat ofcontrol group.The standard and continuous CPR training is elfecfive in improving the quality of CPR operation.

By analysing the anatomy of upper cervical segment, the atlas - axis relation in plain X - ray film in 3-1 cases revealed that the joint displacement is one of the important causes for vertebral artery type or sympathetic type cervical spondylopathy. The main manifestations for diagnosing atlas - axis displacement included headache, dizziness, unilateral protrusion and tenderness of the affected vertebra, displacement of tooth protrusion up to 1 mm at anterior mouth -open positions. The most effective therapy was accurate and dexterousmanipulation for reduction.

Objective To identify the important risk factors for type 2 Diabetes Mellitus (T2DM) and develop effective strategies to address the problem of T2DM.Our study aimed to evaluate the association between apolipoprotein E (ApoE) genetic polymorphism and type 2 diabetes,and to provide clues for the etiology of T2DM.Methods Based on the criteria of inclusion and exclusion,we extracted,pooled,analyzed and assessed the case-control studies of ApoE polymorphism and T2DM published in PubMed,Web of Science,Medline,WanFang,VIP,and CNKI databases by R soft-ware (version 3.4.3).We used Random-effect models when heterogeneity was present in between-study,and fixed-effect models otherwise.Results We had 59 studies covering 6,872 cases with T2DM and 8,250 controls,and compared the alleles and genotypes of ApoE between cases and controls.When we conducted a comparison between ApoE ε4 and ε3 alleles,we produced a pooled OR of 1.18 (95％ CI:1.09-1.28;P ＜ 0.001).ApoE ε2/ε2 genotype displayed a possible association with T2DM (OR =1.46;95％ CI:1.11-1.93;P =0.007),ε3/ε4 genotype showed a 1.11-fold risk (OR =1.11;95％ CI:1.01-1.22;P =0.039) and ε4/ε4 genotype had a 1.71-fold risk of developing T2DM (OR =1.71;95％ CI:1.33-2.19;P ＜ 0.001) when they were compared with ε3/ε3 genotype.Conclusions There is an association between ApoE polymorphism and T2DM:allele ε4 and genotypes (ε2/ε2,ε3/ε4,and ε4/ε4) are associated with the increased risk for the development of T2DM,and they may be risk factors for T2DM.

OBJECTIVETo evaluate perioperative occult blood loss following total knee arthroplasty (TKA).

METHODSA retrospective analysis of 40 patients undergoing TKA was conducted to calculate the mean blood loss and occult blood loss according to Gross formula.

RESULTSThe mean total blood loss was 1538 ml in these cases with occult blood loss of 791 ml. In patients with autologous blood transfusion, the mean total blood loss was 1650 ml with occult blood loss of 786 ml. In patients without autologous blood transfusion, the mean total blood loss was 1370 ml with occult loss of 798 ml.

CONCLUSIONTKA often results in large volume of occult blood loss in the perioperative period which can not be fully compensated by autologous blood transfusion, and additional blood supply is needed for maintenance of the circulating volume.

Aged ; Arthroplasty, Replacement, Knee ; adverse effects ; methods ; Blood Loss, Surgical ; statistics & numerical data ; Blood Transfusion, Autologous ; Female ; Hemoglobins ; analysis ; Humans ; Intraoperative Complications ; blood ; etiology ; Male ; Middle Aged ; Retrospective Studies

The mitochondrial enzyme glutaminase C (GAC) is highly expressed in a variety of cancer cells, resulting in increased glutamine metabolism and cancer development. Therefore, GAC has become a potential target for anti-tumor drug development. However, current GAC inhibitors shared similar structural characteristics, few new scaffolds were reported. By conducting a prokaryotic Escherichia coli expression system, human GAC protein of high-purity was obtained through lysozyme digestion combined with ultrasound dissociation, and cobalt magnetic beads purification, Moreover, we performed studies to validate interaction between small molecules and GAC protein through thermal shift assay, drug affinity responsive target stability assay, protein crosslinking and GAC enzyme activity detection. Meanwhile, a comprehensive small molecule-protein interaction confirmation and systematic pharmacodynamic study in vitro were carried out on compound C19, which was a reported GAC inhibitor screened from the Enamine database. Results showed that C19 directly bind to GAC protein, disturbed GAC tetramers formation, and inhibited its enzyme catalytic activity. By interfering GAC function, C19 dose-dependently suppressed GAC-mediated glutamine metabolism, reduced glutamate in cancer cells, and thus alleviated A549 and NCI-H1299 non-small cell lung cancer cell growth. Together, C19 was identified as a lead compound, providing a new strategy for the structural design of drugs targeting GAC.