Objective To study the effect of neuropeptide on migration of epidermal stem cells in wound repair. Methods 90 new born rats (3～4d old) were randomly divided into three groups: SP group, capsaicin group, and normal group. BrdU labeling, combined with specific protein markers of epidermal stem cells, K19 and ?1 integrin, were used to identify epidermal stem cells. The migration of epidermal stem cells was observed in full thickness skin wound on back on 21 days after injury. Substance P was applied to the skin wound after injury in SP group. The status of migration of epidermal stem cells in SP group was compared with that in capsaicin group, in which capsaicin was injected to destroy sensory neuron before skin injury, and with normal group, in which the skin wound was not treated with any medication. Results Wounds of rats in SP group were healed 18 days after injury. It was shortened by three days compared with normal group. Only 25.54% of wound area was healed in capsaicin group on day 18. There were many epidermal stem cells in the edge of the wound and granulation tissue in Group SP. Only a small number of epidermal stem cells were seen in capsaicin group, where SP release was blocked by chemical destruction of sensory neurons, in the wound edge, but not in granulation tissue. The amount of epidermal stem cells as seen in the wound edge, but not in granulation tissue, in normal group was less than in the SP group but more than in capsaicin group. Conclusions SP obviously promotes wound healing and shortens the healing period. SP can induce epidermal stem cells to migrate into the skin wound edge and granulation tissue.

Acupuncture Therapy ; Analgesia ; Animals ; Brain Chemistry ; Morphine ; pharmacology ; Opium ; analogs & derivatives ; antagonists & inhibitors ; Rats

The study aims to establish a method for simultaneous determination of repaglinide and pravastatin sodium in rat plasma by LC-MS/MS and to study its pharmacokinetic interactions. Eighteen male SD rats were divided into repaglinide group, pravastatin sodium group and co-administration group. Blood samples were collected at different times after oral administration. Repaglinide and pravastatin sodium in rat plasma were separated by Agilent HC-C18 with the mobile phase consisting of methanol-0.1% formic acid (80 : 20). Detection and quantification were performed by using ESI-MS. The detector was operated in selected Reaction-monitoring mode at m/z 453.3-->230.1 for repaglinide, m/z 447.2-->327.4 for pravastatin sodium and m/z 285.1-->192.9 for diazepam as the internal standard. The calibration curve obtained was linear (R2>0.99) over the concentration range of 9.77-10,000 ng.mL-1 for repaglinide and 4.88-625 ng.mL-1 for pravastatin sodium. Compared with the single administration group, Cmax and AUC0-6h of repaglinide increased significantly (P<0.05) and tmax of pravastatin sodium prolonged (P<0.05) in co-administration group. The method is found to be simple, sensitive and accurate for determining the concentration of repaglinide and pravastatin sodium in rat plasma. There exists pharmacokinetic interactions in the co-administration of repaglinide and pravastatin sodium.

OBJECTIVE:To optimize the preparation technology of substance P multivesicular liposomes(Depo-SP)and investigate its property.METHODS:Depo-SP was prepared by multiple emulsion method,with encapsulation efficiency taken as index,molar ratio of phospholipid to cholesterol(A),ultrasound emulsification time(B),volume ratio of colostrum to two-phase water(C)and coemulsifier concentration(D)as factors to design the orthogonal experiment for optimizing the preparation technology.The encapsulation efficiency,distribution of particle size,in vitro drug release profiles,phase transition temperature and melting point of the optimized preparation were determined.RESULTS:The optimized preparation conditions of Depo-SP were as follows:A-2:1,B=30 s,C=1:2 and D=2%.The encapsulation efficiency of the preparation reached 85%;the particle size stood at 0.75～27.75?m;the SP release in normal sodium was t_(1/2)=11 h and the sustained release maintained for 72 hours;the phase transition temperature was 34℃and the melting point was 110℃. CONCLUSION:The prepared Depo-SP had high encapsulation efficiency and stable in physical property,and it is an ideal sustained release preparation.

Objective To screen the non-nucleoside compounds against HIV-1 reverse transcriptase by molecular modeling and bioactivity assay. Methods Surflex-Dock module of Tripos SYBYL software was used to simulate the binding pattern of 22 000 compounds in SPECS database with the active pocket of HIV-1 reverse transcriptase. Based on the simulation results, the interaction mode between the above compounds and the crystal structure of HIV-1 reverse transcriptase was analyzed. The compounds with higher docking scores and better binding pattern were determined by anti-HIV-1 ac tivities test in vitro. Results The virtual screening results showed that the docking conformation of 1- (4-fluorophenyl) -3- [2- (1H-indol-3-yl) ethyl] thiourea was similar to the embedded ligand in Rilpivirine crystal structure. 1- ( 4-fluorophenyl) -3- [ 2- ( 1H-indol-3-yl) ethyl] thiourea was held together with the key residue Lys101 in docking pocket of HIV-1 reverse transcriptase by hydrogen bonds, and hadπ-πstacking action together with the conservative residue Trp229 and the aromatic residue Tyr181 respectively. The bioassay in vitro results showed that when the proliferation rate of C8166 lymphocyte syncytium infected by HIV-1ⅢB arrived 50% ( EC50) , the concentration of 1- ( 4-fluorophenyl) -3- [ 2- ( 1H-indol-3-yl) ethyl] thiourea was 5.45μg/mL. Conclusion Molecule docking technology is an effective approach to reducing the screening of candidate compounds with micromolecular activity, and can be used to predict the interaction mode between the compound and the target receptor. In the study, active compound 1- (4-fluorophenyl) -3- [2- (1H-indol-3-yl) ethyl] thiourea has been screened out by molecule docking technology.

Listeriamonocytogenes is a facultative intracellular bacterium that enters professional antigen presenting cells , presents passenger antigens to the major histocompatibility complex class I and II pathways ,then elicits CD+4 and CD+8 T-cell-mediated immune responses .It was demonstrated that attenuated Listeriamonocytogenes as a novel live vaccine vector in deliv-ering tumor antigens of cervical cancer and melanoma etc .,could induce strong protective immune response ,and shows effec-tive antitumor immunotherapeutics .This review discussed the characteristics of immune responses elicited by Listeria monocy-togenes ,and the progress of its antitumor immunotherapeutics as delivery vaccine vector .

Objective To study the characteristics of craniocerebral injury caused by the handgun bullet impacting on the bulletproof helmet.Methods Fourteen healthy landrace pigs were involved and randomly divided into injury group(n =9)and control group(n =5).The landrace pigs of the injury group were shot vertically on the head under the protection of helmet plate with 9 mm handgun bullet at velocity of 360 m/s.While the landrace pigs of the control group were dealt with the same process as the injury group except for use of unarmed handgun bullet of the same ammunition dose.The changes of vital sign in the early period and the retina injury at two hours after injury were observed.Porcine cerebrospinal fluid (CSF)at pre-injury and at three hours post-injury were obtained for investigating the levels of neuron specific enolase(NSE)andαⅡ-spectrin protein.Then,the landrace pigs were sacrificed and dissected to examine the general morphological changes of the brain.The brain cortex was taken,fixed and stained with hematoxylin-eosin for microscopic observation.Results The landrace pigs in the injury group manifested decrease of the blood pressure and increase of the heart rate and respiratory rate in the early stage after injury.General morphological observation found retinal hemorrhage(3/9),skull fracture(3/9)and brain surface damage including local impact lesion(9/9)and contrecoup lesion(9/9),with severe impact lesion than contrecoup lesion.Optical microscopic observation revealed acute injury of the cerebral cortex neuron both on the impact and contrecoup locations.The concentrations of NSE and αⅡ-spectrinwere significantly increased in CSF three hours after injury(P ＜ 0.05).Conclusions The handgun bullet impacts on the pig head protected by the bulletproof helmet may induce blunt craniocerebral injury in the early period,with severe impact lesion than contrecoup lesion.Therefore,traumatic brain injury of the soldiers armed with the helmets should be stressed and managed early.

The prevalence of human and animal listeriosis for nearly 11 years in China was investigated in this study . The literature information about listeriosis in China from 2002 to 2012 was collected through retrieval system to make clinical and epidemiological statistical analysis of listeriosis .Cases of listeriosis were reported in 27 (79% ) provinces of China .The re-sult showed that animal listeriosis was reported for 123 times ,among these reports ,most were from pigs (39% ) ,and the sheep was in second place .Central nervous system infection was the main clinical manifestation of listeriosis in animals (72% ) . For human listeriosis ,84 clinical cases of listeriosis were reported ,including 35% cases in non-perinatal stage and 65% cases in perinatal stage .The main clinical manifestation of listeriosis was septicemia (51% ) .According to the result of investigation about listeriosis based on literatures information ,Listeriamonocytogenes caused humans and animals listeriosis annually ,which were reported in most provinces of China .The epidemic characteristics for listeriosis suggested that it was essential to strength-en the prevention and control of listeriosis .

Bovine interferon-gamma (BoIFN-gamma) gene was amplified by reverse transcription polymerase chain reaction (RT-PCR) from total RNA of bovine spleen lymphocytes stimulated with ConA. The products of RT-PCR were cloned into pVAX1 vector, positive recombinant clone was identified by restriction enzyme digestion and sequencing. The recombinant plasmid pVAX1-BolFN-gamma was transfected into COS-7 cells mediated by lipofectine, indirect immunofluorescent assay analysis confirmed that rBoIFN-gamma was expressed in COS-7 cells. BoIFN-gamma gene (without signal peptide) was cloned into pET-30a(+) and pGEX-6p-1 vector, and transformed into the Escherichia coli cells. After optimizing the induction condition, SDS-PAGE analysis showed that the expression products were all found in soluble form and had a molecular weight of 23 kDa and 43 kDa respectively. BoLFN-gamma precursor gene (with signal peptide) was cloned into transfer vector pFastBac 1, and transformated into DH10Bac E. coli cells. By site-specific transposition, BoIFN-gamma gene was integrated into shuttle vector Bacmid, and transfected into the Sf9 insect cells mediated by lipofectine to produce recombinant baculovirus. Indirect immunofluorescent assay analysis confirmed that rBac-BoLFN-gamma was expressed successfully in Baculovirus vector system. The antiviral activities of rHis-BoIFN-gamma, rGST-BoIFN-gamma and rBac-BoIFN-gamma were up to 8.389 x10(7) U/mg, 6.554 x10(5) U/mg and 4.096 x 10(4) U/mL respectively, which were analyzed in MDBK/VSV system. A sandwich ELISA was established using monoclonal antibodies 3E6 and 5G4, which can detect BoIFN-gamma in quantity and provide a useful method for the clinical practice and research of BolFN-gamma.
Animals ; Antiviral Agents ; pharmacology ; Baculoviridae ; genetics ; metabolism ; COS Cells ; Cattle ; Cercopithecus aethiops ; Escherichia coli ; genetics ; metabolism ; Genetic Vectors ; genetics ; Interferon-gamma ; biosynthesis ; genetics ; pharmacology ; Recombinant Proteins ; biosynthesis ; genetics ; pharmacology ; Transfection

OBJECTIVETo explore an early stage repair method for soft tissue defect of limbs of modern firearm wound, and to improve treating result.

METHODSDefects of the hind limbs of dogs were repaired with skin, muscle and myocutaneous flaps.

RESULTSWounds healed within 2 weeks in the experimental group except one that healed in 3 weeks because of infection. Limb function was close to normal. The treatment result was better in the experimental group than the control.

CONCLUSIONSSkin, muscle and myocutaneous flaps can cover soft tissue defect at an early stage, prevent and reduce infection, promote the healing and recovery of combined injury, reduce the time of treatment and disability rate.

Animals ; Debridement ; Dogs ; Hindlimb ; injuries ; Microsurgery ; Soft Tissue Injuries ; surgery ; Wound Healing ; Wounds, Gunshot ; surgery