A temperature-sensitive mutant strain was isolated after transposon mutagenesis with Tn5 and named MT54.PCR was carried out with primers designed according to the sequence of transposon,the PCR products showed that the MT54 carried transposon in the genome. The mutant grew well at 30℃in minimal medium(MM)containing p-nitrophenol(PNP)as sole carbon source,while it cannot grow at 37℃in the same medium,NO_2.detection results also proved that.Comparing the degradation rate of PNP and hydroquinone of MT54 and DLL-E4 at different temperature,it was speculated that the mutant site locate in the PNP degradation related genes.

OBJECTIVETo study the synergistic effects of hypothermia and hypoxia on the damage of pulmonary microvascular endothelial cells (PMVEC) in rat.

METHODSPrimary PMVECs were obtained by complex phosphoesterasum digesting from isolated lung tissues of Wistar rats, the PMVECs were identified by phase contrast microscope and immunofluorescence studies for CD31 antigen and bandeiraea simplicifolia isolectin (BSI) binding test. Factorial design was adopted in trial according to hypothermia and hypoxia existing or not. Using corresponding kit measured the levels of lactate dehydrogenase (LDH) activity in cell medium. Level of nitric oxide (NO) concentration was measured by Griess Assay. RT-PCR was used to examine the expression of vascular endothelial growth factor (VEGF), endothelin-1 (ET-1) mRNA in PMVECs.

RESULTSThe monolayer of cultured PMVECs displayed the shape of pavingstone. CD31 antigen and binding BSI results by fluorescence microscope identified the cultured cells were PMVECs. Compared to the control group, LDH activity and VEGF, ET-1 expression levels were significantly increased in hypothermia group, hypoxia group and hypoxia combined with hypothermia group. And the levels of NO concentration were reduced in these three groups. The results of One-way ANOVA showed that there was a synergistic effect between hypothermia and hypoxia.

CONCLUSIONHypothermia and hypoxia both have an effect on PMVECs whether in altering the cell permeability or in releasing of vasoactive substances including NO and ET-1. In addition, there is a synergistic effect between hypothermia and hypoxia.

Animals ; Cell Hypoxia ; Cell Membrane Permeability ; Cells, Cultured ; Cold Temperature ; Endothelial Cells ; cytology ; metabolism ; Endothelin-1 ; metabolism ; Endothelium, Vascular ; cytology ; Lung ; blood supply ; Male ; Nitric Oxide ; metabolism ; RNA, Messenger ; genetics ; Rats ; Rats, Wistar ; Vascular Endothelial Growth Factor A ; metabolism

Objective To evaluate the diagnostic value of the heparin-binding protein (HBP),procalcitonin (PCT),C-reactive protein (CRP),white blood cell (WBC) in respiratory tract bacterial infection.Methods 66 respiratory tract bacterial infection patients,37 respiratory tract non-bacterial infection patients and 39 control group in the Third Xiangya Hospital from October 2015 to March 2017 was selected as objects in this prospective study.The levels of HBP,PCT and CRP in blood of the objects were tested with ELESA,immunofluorescence assay,immunoturbidimetry respectively;WBC counts were taken by Sysmex XE-5000 blood analyzer.The difference among the three groups was analyzed by Student's t test,one-way ANOVA or Wilcoxon test.Receiver operating characteristic curve was utilized to analyze the diagnostic value of HBP,PCT,CRP and WBC in respiratory tract bacterial infection.Results The plasma level of HBP were 36.30 (7.78-89.36) ng/ml,5.57 (4.37-8.23) ng/ml,2.84 (1.53-6.51) ng/ml in respiratory tract bacterial infection group,respiratory tract non-bacterial infection group and control group respectively.The socre of PCT were 0.08 (0.04-0.83) ng/ml,0.09 (0.04-0.30) ng/ml,0.04 (0.03-0.08) ng/ml.The socre of CRP were 56.20 (19.33-76.23) mg/L,34.40 (2.15-83.95) mg/L,(2.20 ± 0.99) mg/L.The socre of WBC count were (10.59 ±4.58) × 109/L,8.40 (5.80-11.88) × 109/L,(6.14± 1.31) × 109/L.There were statistically significant differences in HBP scores between respiratory tract bacterial infection group and respiratory tract non-bacterial infection group or control group (Z =-4.828,P ＜0.001;Z =-5.685,P ＜ 0.001).There were no statistically significant differences in PCT,CRP and WBC scores between respiratory tract bacterial infection group and non-bacterial infection group (F =0.045,P ＞ 0.05;F =0.100,P ＞ 0.05;F =2.417,P ＞ 0.05),but significant differences between respiratory tract bacterial infection group and control group (Z =-2.881,P ＜ 0.05;Z =-6.595,P ＜ 0.001;t =6.499,P ＜ 0.001).The area under curve (AUC) of HBP,PCT,CRP and WBC diagnosing respiratory tract bacterial infection was 0.89,0.69,0.95 and 0.85 respectively.The AUC of HBP differential diagnosising was 0.80.Conclusion HBP can be used as an efficient supplementary indicator for respiratory tract bacterial infection,the differential diagnostic value is superior to PCT,CRP and WBC.

Abstract：Objective To analyze the stabilities of neuraminidase（NA）in influenza vaccine at different temperatures and provide a reference for further complete understanding of overall shelf life of vaccines. Methods Monovalent bulks of influenza H1N1，H3N2 and B vaccines were stored at 4（low temperature），25（room temperature）and 37 ℃（changed temperature）for 0. 5，2，7，24 and 48 h separately，using that at 100 ℃（extreme temperature）for 1 h as control，and determined for NA activity by enzyme⁃linked lectin method. Results The NA activities of influenza H1N1 vaccines stored at 25 and 37 ℃ decreased significantly with the increasing of time. No significant decreases were observed in H3N2 and B vaccines even after storage at two non⁃storage temperatures for 48 h. However，all the NA activities of three vaccines decreased at 100 ℃. Conclusion Both H3N2 and B vaccines showed high stability at abnormal storage temperatures not more than 37 ℃，while H1N1 vaccine was relatively sensitive to the temperature for storage.

OBJECTIVETo explore the damage effects and expression of vascular endothelial growth factor (VEGF) exposed with different low-temperatures on rat dermal microvascular endothelial cells (DMVECs).

METHODSPrimary DMVECs were obtained by discontinuous Percoll gradient centrifugation. The DMVECs were identified by phase contrast microscope and immunofluorescence studies for CD31 antigen. Applied 28 degrees C, 12 degrees C and 0 degrees C to interfere with rat DMVECs as cold-exposure model. The changes of cells morphology were observed under invert microscope. The membrane integrity was determined by lactate dehydrogenase (LDH) activity. RT-PCR was used to examine the expression of vascular endothelial growth factor mRNA in cells.

RESULTSThe monolayer of cultured PMVECs displayed the shape of pavingstone. CD31 antigen and binding BSI results by fluorescence microscope identified the cultured cells were DMVECs. After 24 h cold exposure, the cell morphology of 0 degrees C group was shrunken, the other groups were "Fibroblast-like". The LDH activity (U/L) in the medium of 28 degrees C, 12 degrees C and 0 degrees C groups was 54.17 +/- 3.02, 64.66 +/- 3.03, 82.13 +/- 10.91 respectively, which was significantly higher than that of 37 degrees C group (12.23 +/- 3.0, P < 0.01). The VEGF mRNA expression level was up-regulated in 28 degrees C group and 12 degrees C group versus control group (P < 0.05), but unchanged in 0 degrees C group.

CONCLUSIONThe rat DMVECs injury severity are deteriorated with temperature decreasing, and VEGF might be involved in the regulation of membrane permeability in this period.

Animals ; Animals, Newborn ; Cells, Cultured ; Cold Temperature ; Dermis ; blood supply ; Endothelial Cells ; cytology ; metabolism ; Endothelium, Vascular ; cytology ; Rats ; Rats, Wistar ; Vascular Endothelial Growth Factor A ; metabolism

OBJECTIVETo investigate the protective effects of natakalim against rat aortic vascular endothelial cells (RAVECs) injuries induced by hypoxia and its mechanisms.

METHODSSelecting RAVECs as a cell model injured by hypoxia, these RAVECs were divided into 5 groups: i.e. control group, hypoxia group, natakalim low, medium and high group. The cell survival rate was determined by MTT assay, con was measured using Griess Assay, RT-PCR was used to examine t he expression of intercellular adhesion molecule-1 (ICAM-1), vascular endothelial growth factor (VEGF), endothelin-1 (ET-1) mRNA in RAVEC.

RESULTSNatakalim could reverse hypoxia-induced changes in endothelial cell function, including increased endothelial cell survival rate and level of NO concentration, significantly inhibited the hypoxia-induced endothelial ICAM-1, ET-1, VEGF mRNA expression levels increased.

CONCLUSIONNatakalim have protective effects on hypoxia-induced changes in endothelial cell function, increasing of permeation, excess expression of cell adhesion molecules.

Allyl Compounds ; pharmacology ; Animals ; Aorta ; cytology ; metabolism ; Cell Hypoxia ; Cells, Cultured ; Endothelial Cells ; metabolism ; Endothelin-1 ; metabolism ; Intercellular Adhesion Molecule-1 ; metabolism ; Male ; Propylamines ; pharmacology ; RNA, Messenger ; genetics ; Rats ; Rats, Wistar ; Vascular Endothelial Growth Factor A ; metabolism ; Vascular System Injuries ; metabolism

OBJECTIVETo study protective effect and pathogenesis of complex salvia miltiorrhiza (DanShen) on acute mercury poisoning in rabbits.

METHODSModels of acute mercury poisoning was made in rabbits. The effect of complex salvia miltiorrhiza on blood urea nitrogen (BUN), copper-protein (CP), lactate dehydrogenase (LDH), acid phosphatase (ACP) and the malondialdehyde (MDA) in plasma, superoxide dismutase (SOD) in erythrocyte and MDA, SOD in tissues homogenate were observed.

RESULTSThe administration of complex salvia miltiorrhiza after mercury injection 0.5 h and 9.5 h, decreased BUN, CP, MDA, LDH and ACP, and prevented the reduction of SOD. Compared with mercury poisoning group, the difference was statistical significant (P < 0.05, P < 0.01).

CONCLUSIONIt is suggested that acute mercury poisoning may result in renal damage but also multiple organ tissues, and complex salvia miltiorrhiza possesses protective effect, through stabilized membranes.

Animals ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Female ; Male ; Mercury Poisoning ; blood ; drug therapy ; Phenanthrolines ; pharmacology ; therapeutic use ; Rabbits ; Salvia miltiorrhiza

OBJECTIVETo explore the effect of auricular points sticking-pressing (APSP) in treating post-cesarean hypogalactia (PCH).

METHODSA randomized, controlled, single-blinded clinical trial on 116 patients with PCH was carried out. They were equally assigned to the treatment group and the control group. The treatment group received APSP, with the pellets pressed for 4 times daily, while the control group was only asked to do lactation to meet infant demand. The therapeutic efficacy and the changes in scores of traditional Chinese medicine (TCM) syndrome, volume of milk secretion, supplementary feeding and serum level of prolactin (PRL) in the two groups were estimated and compared after the patients had been treated for 5 days.

RESULTSThe cured and markedly effective rate in the treatment group was 89.7%, which was significantly higher than that in the control group (27.6%, P<0.05), 95% CI (0.1543, 0.2527). The improvement of TCM syndrome, elevation of milking volume, decrease of the supplementary feeding and increase of PRL level revealed in the treatment group were all superior to those in the control group, showing statistical significance (P<0.01).

CONCLUSIONAPSP shows an apparent efficacy in treating PCH and is worthy of application in clinical practice.

Acupuncture Points ; Acupuncture, Ear ; adverse effects ; methods ; Adult ; Cesarean Section ; adverse effects ; rehabilitation ; Female ; Humans ; Infant Nutritional Physiological Phenomena ; Infant, Newborn ; Lactation ; physiology ; Lactation Disorders ; blood ; therapy ; Medicine, Chinese Traditional ; adverse effects ; methods ; Milk, Human ; secretion ; Postoperative Complications ; therapy ; Pregnancy ; Pressure ; Prolactin ; blood ; Treatment Outcome ; Young Adult

This study was aimed to investigate the expression of Hippo signaling pathway core element MST1 gene in acute leukemia (AL) patients, and explore its relation with AL pathogenesis and prognosis. 50 newly diagnosed patients with AL, 33 normal people, 23 patients with AL in complete remission, 12 refractory or relapsed patients were tested for the expression of MST1 gene by real-time quantitative PCR, Western blot was used to further validate the level of MST1 protein expression. And combined with clinical data, prognostic factors of the patients were analyzed. The results indicated that compared with the normal people, the expression level of MST1 in newly diagnosed patients with AL significantly decreased (P < 0.05), but significantly increased in AL patients with complete remission (CR), the difference of expression was statistically significant before CR and after CR (P < 0.05). Compared with refractory or relapsed patients, the expression level of MST1 gene in newly diagnosed patients was not significantly different (P > 0.05). Besides, the expression level of MST1 between the patients with CR and the normal people was not significantly different (P > 0.05). It is concluded that the low expression of MST1 may be related with the pathogenesis and prognosis of AL.
Acute Disease ; Adolescent ; Adult ; Aged ; Aged, 80 and over ; Child ; Child, Preschool ; Female ; Gene Expression Regulation, Leukemic ; Humans ; Leukemia ; diagnosis ; metabolism ; pathology ; Male ; Middle Aged ; Prognosis ; Protein-Serine-Threonine Kinases ; metabolism ; Signal Transduction ; Young Adult