The cytolytic effect of perforin is a mechanism of anti-virus,killing microbial-infected cells and tumor cells.Perforin is a very important non-specific immune factors in fish.In order to understand the function of perforin,the cDNA of grass carp perforin C-terminal peptide was amplified from grass carp liver and kidney cDNA library.It contains a protein kinase C conserved region 2(C2).The cDNA was connected with pET32a,and transformed to expression bacteria DE3.PFP-C was expressed by a prokaryotic expression system and then purified by affinity chromatography.It showed a significant haemolytic activity when tested with rabbit red cells,the optimal pH for haemolytic activity was 7.5,and its haemolytic function dependents on Ca2+ apparently.

Objective To exPlore the effect of mammalian target of raPamycin ( mTOR ) inhibitor eVerolimus on radiosensitiVity of human non_small cell lung cancer cell line in vitro by using eVerolimus to inhibit mTOR signaling Pathway of A549. Methods Human non_small cell lung cancer cell line A549 was subjected to radiation alone or in combination with eVerolimus treatment. The 50%inhibition concentration ( IC50 ) of eVerolimus in A549 cells was detected by methylthiazol tetrazolium ( MTT) assay in vitro. EVerolimus at the 20%inhibition concentration ( IC20 ) was used to Pretreat A549 cells for 24 h. Cells were then irradiated by X_ray with 2,4,6,8 Gy. The cell surViVal fraction was comPuted by clone formation. Cell surViVal curVe was fitted by multitarget one_hit model, and mean lethal dose ( D0 ), dose quasithreshold ( Dq ), surViVal fraction at 2 Gy ( SF2 ), and sensitization enhancement ratio (SER) were calculated. The exPression ofγ_H2AX was determined by Western blotting and then the relatiVe gray Values were analyzed. Results EVerolimus significantly imProVed the sensitiVity of A549 cells to radiation. The D0 , Dq and SF2 of eVerolimus+irradiation grouP were significantly lower than those of irradiation grouP. The SER was 1. 36. The residual amount of γ_H2AX Protein in the eVerolimus + irradiation grouP was significantly higher than that of the irradiation grouP. Conclusion EVerolimus inhibiting mTOR signaling Pathway can increase the radiosensitiVity of A549 cells.

A large body of research including animal and human studies has confirmed the crucial role of the serotonin (5-HT) system in the regulation of nociception and chronic pain-related behaviors. In recent years, the functional status of the 5-HT system in descending inhibition and facilitation of spinal nociceptive processing has been reevaluated by novel genetic manipulation techniques and selective agents for 5-HT receptor subtypes. Although these studies shed more light on several aspects of descending 5-HT and spinal 5-HT receptors functioning in descending modulation in pain perception, the current knowledge about the specific role of descending 5-HT system in the induction and maintenance of persistent pain remains fragmentary. In this paper, we review the available data from recent studies of the inhibitory or facilitatory influence from descending 5-HT-spinal 5-HT receptor system in acute and persistent pain, attempt to dissect the involvement of this signaling pathway in neural circuits of maintenance of persistent pain and discuss some issues that need to be considered for further pain research.
Animals ; Humans ; Pain ; physiopathology ; Receptors, Serotonin ; physiology ; Serotonin ; physiology

In vivo extracellular recordings were made in the subthalamic nucleus (STN) of intact control rats and rats with 5,7-dihydroxytryptamine (5,7-DHT) -produced lesion of dorsal raphe nucleus (DRN). The results showed that the firing rate of STN neurons in control rats and DRN-lesioned rats were (6.93+/-6.55) Hz and (11.27+/-9.31) Hz, respectively, and the firing rate of DRN-lesioned rats significantly increased when compared to the control rats (P<0.01). In control rats, 13% of STN neurons discharged regularly, 46% irregularly and 41% in bursts. In DRN-lesioned rats, 9% of STN neurons discharged regularly, 14% irregularly and 77% in bursts, the percentage of STN neurons firing in bursts was obviously higher than that of the control rats (P<0.01). In addition, the mean interspike interval coefficient of variation of STN neurons in control rats and DRN-lesioned rats were (0.05+/-0.04) and (0.11+/-0.09), respectively. The mean interspike interval coefficient of variation of DRN-lesioned rats was significantly higher than that of the control rats (P<0.001). These results show that the firing rate and the bursting pattern rate of neurons in STN of DRN-lesioned rats increase significantly, suggesting that DRN inhibits the neuronal activity of the subthalamic neurons in the intact rat.
5,7-Dihydroxytryptamine ; pharmacology ; Adrenergic Agents ; pharmacology ; Animals ; Electrophysiological Phenomena ; Male ; Neurons ; physiology ; Random Allocation ; Raphe Nuclei ; drug effects ; pathology ; Rats ; Rats, Sprague-Dawley ; Subthalamic Nucleus ; physiopathology

In the present study, changes of the neuronal activity of 5-hydroxytrypamine (5-HT) neurons of dorsal raphe nucleus(DRN) in a rat model of Parkinson's disease (PD) were investigated with glass microelectrode recording. The results showed that the discharge rates of 5-HT neurons in control and PD rats were (1.61+/-0.56) Hz and (2.61+/-1.97) Hz, respectively. The discharge rate of PD rats was significantly increased when compared to that of the control rats. In control rats, 79% of 5-HT neurons discharged regularly and 21% in bursts. In PD rats, however, 36% of 5-HT neurons discharged regularly, 16% irregularly and 47% in bursts. The percentage of 5-HT neurons discharging in bursts was obviously higher than that of the control rats (P<0.05). The data suggest that the discharge rate and bursting pattern of 5-HT neurons in DRN are increased in a rat model of Parkinson's disease.
Animals ; Electrophysiology ; Male ; Microelectrodes ; Neurons ; physiology ; Parkinson Disease ; physiopathology ; Raphe Nuclei ; metabolism ; physiopathology ; Rats ; Rats, Sprague-Dawley ; Serotonin ; metabolism

OBJECTIVETo investigate the analgesic and sedative effects of inhaling a mixture of nitrous oxide and oxygen on burn patient during and after dressing change.

METHODSA total of 240 burn patients hospitalized in the Institute of Burn Research of Changhai Hospital Affiliated to the Second Military Medical University, Department of Burns of the First People's Hospital in Zhengzhou, and Department of Burns and Plastic Surgery of General Hospital of Ningxia Medical University from October 2011 to September 2012 were enrolled in our study, and they were all in accordance with the inclusion criteria. The 240 patients were divided into control group (n = 60, treated with inhalation of oxygen during dressing change) and treatment group (n = 180, treated with inhalation of a mixture of 65% nitrous oxide and oxygen during dressing change) according to the computer-generated list of random number. The other treatments in control group and treatment group were the same. Before, during, and after dressing change, heart rate (HR), systolic blood pressure (SBP), diastolic blood pressure (DBP), oxygen saturation (SO2), and adverse effects were observed. The degree of pain and anxiety felt by the patients were respectively evaluated with the visual analogue scale (VAS) and Chinese version of the burn specific pain anxiety scale (C-BSPAS) at the same time points as above. Data were processed with analysis of covariance, chi-square test, analysis of variance, and rank sum test.

RESULTSThere were no significant differences between control group and treatment group in the levels of HR, SBP, DBP, and SO2 before dressing change (with F values respectively 0.76, 0.06, 1.11, 0.70, P values all above 0.05). Compared with those of control group, the levels of HR, SBP, DBP, and SO2 in treatment group were significantly ameliorated during dressing change (with F values respectively 81.78, 146.36, 226.44, 205.62, P values all below 0.01). After dressing change, the levels of DBP in the two groups were close (F = 0.31, P > 0.05), but the levels of HR, SBP, and SO2 showed statistical differences (with F values respectively 7.02, 8.69, 12.23, P < 0.05 or P < 0.01). Before dressing change, the VAS scores were approximate between control group and treatment group (Z = 0.21, P > 0.05). Compared with those in control group (9.4 ± 0.7, 1.7 ± 2.5), the VAS scores were significantly lowered in treatment group during and after dressing change (1.6 ± 1.3, 0.7 ± 1.1, with Z values respectively 11.84, 3.35, P values all below 0.01). There was no significant difference in C-BSPAS score between control group and treatment group before dressing change (Z = 0.62, P > 0.05). Compared with those in control group (75 ± 13, 73 ± 12), the C-BSPAS scores in treatment group were decreased during and after dressing change (9 ± 15, 9 ± 14, with Z values respectively 11.91, 12.28, P values all below 0.01). There were no obvious adverse effects in two groups before, during, and after dressing change.

CONCLUSIONSA mixture of nitrous oxide and oxygen seems to have obvious analgesic and sedative effects on burn patients during dressing change, and it can be widely used.

Administration, Inhalation ; Adolescent ; Adult ; Aged ; Analgesia ; methods ; Bandages ; Burns ; surgery ; Female ; Humans ; Hypnotics and Sedatives ; administration & dosage ; therapeutic use ; Male ; Middle Aged ; Nitrous Oxide ; administration & dosage ; therapeutic use ; Oxygen ; administration & dosage ; therapeutic use ; Young Adult

OBJECTIVETo evaluate color Doppler ultrasonography (CDU) appearances of renal vein thrombosis (RVT) and its diagnostic value.

METHODSTen patients with RVT were analyzed retrospectively. Renal structure, distributions of intrarenal flow signals, echogenicity, and flow fullness in main renal veins were observed with CDU. Resistance index (RI) was recorded from the waveforms of segmental or interlobar renal artery.

RESULTSTen kidneys in nine patients were confirmed to have thrombus within the main renal veins, and one patient was confirmed to have thrombus within the small intrarenal veins. The appearances of the main renal vein thrombosis included full of solid echogenicity or strip echogenicity and complete or partial filling defect within the main renal veins, and absent or a few intrarenal venous flow signals in 70% of kidneys involved. The appearances of intrarenal vein thrombosis included obscure renal structure and no venous flow signal within the involved part of the kidneys. Reverse diastolic flow in the intrarenal artery had only a sensitivity of 36% (4/11); in other 7 kidneys without intrarenal arterial reverse diastolic flow, increased RI (mean, 0.84; range, 0.74-0.96) was found.

CONCLUSIONCDU is helpful for rapid clinical diagnosis and follow-up of RVT, and therefore can be the first imaging modality of choice for RVT.

Humans ; Renal Circulation ; Renal Veins ; diagnostic imaging ; Retrospective Studies ; Thrombolytic Therapy ; Ultrasonography, Doppler, Color ; methods ; Venous Thrombosis ; diagnostic imaging ; drug therapy

OBJECTIVETo search for an effective method of reducing intraoperative blood loss in radical retropubic prostatectomy (RRP).

METHODSWe performed RRP for 100 patients with prostate cancer, 50 (group A) with the Walsh or Poor method for handling the dorsal venous complex (DVC), and the other 50 (group B) through the following three additional procedures for hemostasis: first placing a #7 prophylactic suture in the distal position of DVC, then ligating the vascular bundle of the prostatic apex with continuous 4-0 Vicryl sutures, and lastly placing a 4-0 absorbable suture followed by freeing the neurovascular bundle (NVB) or freeing NVB before suturing the remained levator ani myofascia and the deep layer of Denovilliers' fascia above the rectal serosa with 4-0 Vicryl. We assessed the effects of the three hemostatic methods in RRP by comparing the volumes of intraoperative blood loss and transfusion, operation time and perioperative levels of hemoglobin.

RESULTSThere were no significant differences between groups A and B in age, PSA, Gleason score, clinical stage, prostate volume, operation time and perioperative hemoglobin levels (P>0.05). The volumes of intraoperative blood loss and transfusion were markedly higher in group A ([1103.00 +/- 528.03] ml and [482.00 +/- 364.60] ml) than in B ([528.00 +/- 258.96] ml and [140.00 +/- 266.28] ml) (P<0.05).

CONCLUSIONIntraoperative blood loss in RRP could be significantly decreased by placing a prophylactic hemostatic suture in the distal position of DVC, continuous suture of the vascular bundle of the prostatic apex after cutting off the urethra, and placing a fine absorbable suture above NVB or continuous suture of the remained levator ani mony fascia and the deep layer of Denovilliers'fascia above the rectal serosa with absorbable sutures after freeing NVB.

Aged ; Blood Loss, Surgical ; prevention & control ; Hemostatic Techniques ; Humans ; Male ; Middle Aged ; Prostatectomy ; methods ; Prostatic Neoplasms ; surgery

OBJECTIVEInvestigations were carried out to understand the effect of 50 Hz power frequency magnetic field on microfilament assembly of human amniotic cells and on expression of actin and epidermal growth factor receptor.

METHODSHuman amnion FL cells were exposed to 0.1, 0.2, 0.3, 0.4, 0.5 mT power frequency magnetic field for 30 minutes. Microfilaments were marked using Phalloidin-TRITC, and then were observed under a fluorescence microscope. An optical method was used to detect the relative content of microfilament in cells. A scanning electron microscope was used to detect the cell shape. The content of actin and epidermal growth factor receptor in the preparation of the detergent-insoluble cytoskeleton were measured by western-blotting to analyse the potential mechanism of the change induced by magnetic field.

RESULTSIntracellular stress fibers were found to decrease after exposing cells to a 0.2 mT power frequency magnetic field for 30 minutes. New microfilament and filopodia bundles appeared at the cell periphery after exposure, but the detected total F-actin content per cell was not significantly changed, detected by a F-actin-specific dye. The change in the amount of microfilaments caused by the field could be recovered 2 hours later when the field was withdrawn. The mean height of microfilament cytoskeleton decreased from (12.37 +/- 1.28) microm to (9.97 +/- 0.38) microm (t = 6.96, P > 0.05) after exposure using a confocal microscope. The cell shapes became more flat and lamellipodia appeared after exposure observed by a scanning electron microscope. By using Western blotting method, the intracellular contents of epidermal growth factor receptor and of actin in the preparation of the detergent-insoluble cytoskeleton which are associated with high-affinity epidermal growth factor receptors, increased about (31.2 +/- 4.1)% (t = 17.10, P < 0.05) and (16.8 +/- 2.3)% (t = 16.68, P < 0.05) respectively, compared with that of the control.

CONCLUSIONThese results suggest that a short time exposure to a 0.2 mT power frequency magnetic field induces re-organization of microfilament in human amnion FL cells. These changes could be recovered by field withdraw and may have something with the clustering of epidermal growth factor receptors induced by magnetic field.

Actin Cytoskeleton ; metabolism ; Amnion ; cytology ; radiation effects ; Cell Line ; Cell Movement ; Cytoskeleton ; metabolism ; radiation effects ; Electromagnetic Fields ; Humans ; Receptor, Epidermal Growth Factor ; metabolism ; Signal Transduction

AIMGenetic diversity, relationship and molecular authentication of total 8 wild populations of Dendrobium officinale were investigated using RAPD markers.

METHODS10 random decamer primers were screened for Random Amplified Polymophic DNA (RAPD) fragments. A DNA molecular dendrogram was established based on cluster analysis by UPGMA (unweighted pair-group method with arithmetic average), and the relationship of the wild populations were analyzed, and all the wild populations were authenticated.

RESULTSA total of 439 loci with an average of 43.9 loci per primer and 54.9 loci per population were amplified from 8 wild populations by 10 effective primers. In the total 104 amplified bands, 95 were polymorphic, corresponding to 91.35% genetic polymorphism. The genetic distances were 0. 590 to 0. 727, with an average of 0. 686.

CONCLUSIONDistinct genetic differences and extensive genetic diversity were presented among the wild populations. RAPD markers were an informative and useful tool for the genetic diversity, evaluation and authentication of wild populations of Dendrobium officinale. Primer S412 could be used to authenticate 8 wild populations completely.

China ; Cluster Analysis ; DNA Fingerprinting ; DNA Primers ; DNA, Plant ; genetics ; Dendrobium ; genetics ; Ecosystem ; Genetic Markers ; Genetic Variation ; Phylogeny ; Plants, Medicinal ; genetics ; Random Amplified Polymorphic DNA Technique