In order to evaluate the changes in T cell subsets after acute irradiation injury, and investigate the mechanism of deficiency of cellular immunity in acute irradiation syndrome, Balb/c mice were exposed to a single total body irradiation of 5Gy gamma rays from a 60 Co source. On days 10, 28 and 120, the amounts of CD4 + and CD8 + subsets in spleen cells were evaluated by flow cytometry (FCM), and expression of IL 4 gene in ConA stimulated spleen cells was evaluated by reverse transcription linked polymerase chain reaction, in which ?-actin gene was used as an internal control. It was found that after irradiation, the ratio of CD4 + /CD8 + became significantly higher than that of the controls, and then the ratio lowered gradually until day 120. However, it still did not return to the level of the controls. On day 10, expression of IL 4 gene at mRNA level increased significantly. The results indicated that CD8 + subset was more radiosensitive than CD4 + , and the pattern of cytokine seeretion shifted to that of Th2 soon after irradiation.

BACKGROUND:Previous study has found that hsa-miR-182 is probably related to the apoptosis-related genes such as cytochrome C (Cycs C) and calcineurin subunit CnB (PPP3R1) in nucleus pulposus cells.
OBJECTIVE:To determine whether miR-182 plays a regulatory role in nucleus pulposus cel apoptosis by detecting the relative gene expression levels after transfecting miR-182 with Cycs C and PPP3R1 into nucleus pulposus cel s via plasmid delivery.
METHODS:After a bioinformatics prediction about miR-182, miR-182 and target genes were transfected into the nucleus pulposus cel s, and at the same time, blank control group was established. Then the expression levels of the target genes were detected through cel lysis.
RESULTS AND CONCLUSION:miR-182 significantly inhibited the expression of Cycs C in nucleus pulposus cel s compared with the blank control group (P<0.05). Compared with the blank control group, miR-182 made no inhibitory effect on the expression of PPP3R1. These findings suggest that miR-182 may play a regulatory part in nucleus pulposus cel apoptosis by inhibiting the expression of Cycs C.

Biocompatible Materials ; analysis ; Chromatography ; methods ; Chromatography, Gas ; methods ; Chromatography, Liquid ; methods ; Paraquat ; analysis ; Pesticide Residues ; analysis

Animals ; Brain ; pathology ; Hydroxyproline ; metabolism ; Kidney ; pathology ; Liver ; pathology ; Lung ; metabolism ; pathology ; Male ; Paraquat ; metabolism ; poisoning ; Rats ; Rats, Wistar

BACKGROUND:We have designed and manufactured a novel artificial cervical vertebra and intervertebral complex (ACVC) which combines the cervical titanium cage with the artificial cervical disc, and also developed the ACVC with a hydroxyapatite biocoating (ACVC-HA). OBJECTIVE:To evaluate biomechanical properties of the joint system, and the role of HA coating in promoting osseointegration and long-term stability. METHODS:Twenty-four goats were randomly divided into three groups and underwent the anterior C2/3 and C3/4 discectomy, and C3 subtotal corpectomy, fol owed by ACVC implantation (group 1) and ACVC-HA implantation (group 2), and given no intervention (black control group), respectively. group. At 12 weeks after surgery, C1-5 samples were col ected to undergo biomechanical tests and histological staining. RESULTS AND CONCLUSION:Prior to the fatigue test, compared with the blank control group, the range of motion and neural zone of groups 1 and 2 in the directions of flexion-extension and lateral bending showed no significant differences, but the above indicators were significantly increased in the direction of rotation (P<0.05). Additional y, the stiffness in al three directions was significantly lower than that in the blank control group (P<0.05). There were no significant differences in the range of motion and neural zone in al directions between groups 1 and 2. Similar results were found after the fatigue test. The histological staining showed that both two implants had good biocompatibility and abradability, but more new bone formed on the ACVC-HA. These results suggest that ACVC can effectively reconstruct the motor function of the cervical spine after decompression. Furthermore, HA coating can markedly improve bone-implant interface to promote osseointegration.

Objective Toinvestigate the role of dynamic dispersion of electrical recovery in ventricular fibrillation maintenance mechanism.Method Thirty-seren male Yorkshire pigs weighing (23?2.5) kg at 3 months age were randomly divided into Sham-operated group and myocardial infarction group in which the first branch of left anterior descending coronary artery of animals was hgated.After 4 weeks,according to VF occurrence by programmed electrical simulation,the myocardial infarction hearts were divided into two groups, twelve hearts in VF (+) group or six hearts in VF (-) group;and the normal hearts were divided into two groups,five hearts in VF (+) group or ten hearts in VF (-) group.Action potential duration (APD) of subendocardial,subepicardial and mid-layer myocardium of left ventricular anterior wall were recorded simultaneously.The relationship between APD and diastolic interval was quantified as an electrical restitution curve.A Student's-test was used to specify differences between groups.Results At the fourth week after coronary artery ligation,eighteen myocardial infarction hearts and fifteen normal hearts were investigated.When investigating transmural dispersion of reporlarization (TDR) of left ventricle,there was no difference between VF (+) group and VF (-) group [ (20.43?7.01) ms vs (22.32?7.53) ms,P=0.45],although there was statistically significant between normal hearts and myocardial infarction hearts [ (15.66?4.45) ms vs (25.72?6.70) ms,P =0.001].The percentage of APD restitution slope exceed 1 and APD restitution slope maximum were high in the VF (+) group with myocardial infarction hearts and in the VF (+) group with normal hearts, lower in the the VF (-) with normal hearts and lowest in the the VF (-) group with myocardial infarction hearts.Conclusion Dynamic APD restitution properties may play an important role in the ventricular fibrillation maintenance mechanism.The APD restitution slope and not static TDR have prognostic value regarding ventricular fibrillation events.The hearts with the steepest of APD restitution curves has the greatest liability of ventricular fibrillation to happen.

BACKGROUND:Development of new coronary thrombolytic agents is hot in the market. A new drug, mutated recombinant tissue-type plasminogen activator (rtPAm), is the product of mutation of tPA by changing binding loci with plasminogen activator inhibitor (PAI)-1 to reduce the degradation. In vitro test has demonstrated that the activity of rtPAm is much higher than rtPA in the absence of PAI. The present study is to observe the efficacy of mutated recombinant tissue-type plasminogen activator (rtPAm) in coronary thrombolytic therapy. METHODS:A total of 30 adult beagles were equally divided into 5 groups after thrombi:vehicle group, urokinase group, rtPAm low-dose group, rtPAm medium-dose group, and rtPAm high-dose group. Thrombolytic effect and myocardial infarction were observed after thrombolytic therapy. RESULTS:In the urokinase group, time to reperfusion was (15.8±3.8) minutes. TIMI 2 flow was demonstrated in 4 beagles, TIMI 3 flow in 2, and re-occlusion in 4 after 90 minutes respectively. In the low-dose rtPAm group, time to reperfusion was (15±4.5) minutes; TIMI 2 flow was demonstrated in 2 beagles, TIMI 3 flow in 4, and re-occlusion in 2 after 90 minutes. In the high-dose rtPAm group, time to reperfusion was (7.5±2.6) minutes. None of the beagles showed re-occlusion after 90 minutes. The infarction areas were (2.1+0.9)％ in the medium-dose rtPAm group and (0.7+0.4)％ in the high-dose rtPAm group, which decreased significantly than those in the low-dose rtPAm group. The aggregation rate in the medium-dose and high-dose rtPAm groups decreased significantly than that in the urokinase group. CONCLUSION:rtPAm may serve as a thrombolytic agent with platelet-targeted fibrinolysis and antiplatelet aggregation activities.

ABSTRACT:In the present study ,we aimed to identify differentially expressed proteins between induced worms (the infec‐ted mice were treated intragastrically with ED50 PZQ) and uninduced worms (control group) for clarifying the mechanism of PZQ .ED50 PZQ was used to administrate mice that were infected with S .japonicum via intragastric incubation for consecutive‐ly 30 days .Twenty‐one days later ,mice were sacrificed after treatment with 200 mg/kg PZQ for continuously five days ,and the male worms were obtained and some of them were subjected in DMEM medium with different concentrations of PZQ in vitro for 16 hours .Then the worms were washed twice and incubated in PZQ‐free medium for 72 hours .Compared with control group ,the induced worms had lesser sensitivity to PZQ .The survival rate of induced worms was 75 .6% in vitro when the con‐centration of PZQ was 112 mol/L (the concentration was 8 times of uninduced worms Lethal Concentration ) ,significantly higher than that in the uninduced worms (11 .1% ,P<0 .05) ,showing obviously tolerance .The other induced and uninduced worms were acquired and collected for 2D‐DIGE and MALDI‐TOF‐MS ,and combined with bioinformatics to analyse the func‐tion of the identified protein .Thirty differential expression proteins were confirmed between induced and uninduced worms ,in‐cluding 12 proteins up‐regulated and 18 proteins down‐regulated .These proteins respectively ascribed to cytoskeleton‐associat‐ed protein ,glucose and energy metabolism enzymes ,stress proteins ,thioredoxin peroxidase enzymes ,and other protease .Up‐or down‐regulation of these differential proteins indicated that PZQ promote or inhibit the expression of some specific genes . These findings may help to clarify the mechanism of PZQ ,simultaneously ,providing a scientific basis for exploring new vaccine candidate antigens and targets for drug therapy .

AIM: To verify and localize the expression of nicotinamide N-methyltransferase (NNMT) in pancreas of streptozotocin(STZ)-induced diabetic monkeys and understand its important role in ?-cell destruction in the pathogenesis of diabetes. METHODS: Through an olig-microarray gene chip, NNMT was identified as the most obviously up-regulated genes in pancreas of STZ-induced diabetic monkeys versus controls. Semiquantitative RT-PCR and Western blotting were performed to verify the differential expression at mRNA and protein level respectively. Then the cellular localization of NNMT expression within pancreas was identified by immunohistochemical(IHC) staining.RESULTS: An obvious high expression of NNMT at both mRNA and protein levels was shown in pancreas of STZ-induced diabetic monkeys compared to that of controls. Further localization of the protein by IHC staining in pancreas specimens showed that its altered expression was restricted to central islets, most of which were ? cells.CONCLUSION: Expression of NNMT is increased in islets of STZ- induced diabetic monkeys, which infers that NNMT might participate in the process of ? cell damage in diabetes probably through the mechanism of energy metabolism disturbance.

OBJECTIVETo investigate the effect of salidroside on hypoxia-induced apoptosis of corpus cavernosum smooth muscle cells (CCSMCs) in rats.

METHODSRat CCSMCs were cultured in vitro by the enzyme digestion method and identified by immunofluorescent staining of anti-alpha-SMA and anti-Desmin. The non-toxic dose of salidroside was determined by MTT assay. Low-oxygen mixed gas (1% O2, 5% CO2, and 94% N2) was piped into a modular incubator chamber to induce hypoxia. The CCSMCs were divided into a normal, a hypoxia, and a 32 microg/mL salidroside intervention group. The apoptosis of the CCSMCs was detected by flow cytometry and the expression of the caspase-3 protein determined by Western blot.

RESULTSThe majority of the CCSMCs were positive for alpha-SMA and Desmin at immunofluorescent staining. Salidroside at < 32 microg/ml produced no obvious toxicity to CCSMCs. Compared with the normal control group, the rates of early and late apoptosis of CCSMCs were both increased significantly in the hypoxia group ([12.77 +/-1.41]% vs [18.69 +/- 1.29]%, P < 0.01 and [14.63 +/- 2.00]% vs [21.03 +/- 1.530]% , P < 0.05). Western blot showed a markedly increased expression of cleaved caspase-3 (P < 0.01). Intervention with 32 microg/ml salidroside significantly reduced hypoxia-induced early apoptosis of CCSMCs ([13.46% +/- 1.87]%, P < 0.01) and decreased the expression of cleaved caspase-3 (P < 0.01).

CONCLUSIONSalidroside can reduce the expression of cleaved caspase-3 and inhibit hypoxia-induced apoptosis of CCSMCs in rats.

Animals ; Apoptosis ; drug effects ; physiology ; Caspase 3 ; metabolism ; Cell Hypoxia ; physiology ; Cells, Cultured ; Glucosides ; pharmacology ; Humans ; Male ; Myocytes, Smooth Muscle ; cytology ; drug effects ; enzymology ; Penis ; cytology ; drug effects ; Phenols ; pharmacology ; Rats