Objective To investigate the role of induction of carbon monoxide (CO) with methylene chloride (MC) in recipients in ameliorating allograft rejection and prolonging allograft survival and to explore the possible mechanisms in a murine heart transplantation model.Methods Inbred male C57BL/6 mice were used as donors and inbred male Balb/c mice as recipients respectively to establish cervical heterotopic heart transplantation model.The experiments were divided into 3 groups.Recipients were treated with MC (100 mg/kg,per os) day 1 prior to transplantation to day 6 posttransplantation (group MC1w) or to day 13 posttransplantation (group MC2w),or treated with isovolumic olive oil day 1 prior to transplantation to cardiac arrest of allograft (group Tx).The serum TNF-α and IL-10 proteins,TNF-α and IL-10 mRNA,and Foxp3 mRNA and protein in cardiac grafts were measured respectively.The intercellular adhesion molecule-1 (ICAM-1) and Caspase-3 protein in cardiac grafts,and the histopathologic changes of cardiac grafts were also observed.Results Serum COHb levels in untreated mice were (0.85 ± 0.28)％.After MC application,serum COHb peaked within 3 h in recipients (5.24 ± 0,45)％ (P＜0.01 ).The median survival time of cardiac grafts in group MC1w(12.1 days) and group MC2w( 19.4 days) was longer than that in group Tx (6.3 days) (P ＜0.01). As compared with group Tx,induction of CO in group MC1w and group MC2w down-regulated significantly the levels of serum TNF-α (P＜0.01 ) and TNF-α mRNA (P＜0.01) of cardiac grafts and spleen in recipient mice,inhibited the protein expression of ICAM-1 (P＜0.01) and Caspase-3 (P＜0.01) of cardiac grafts,and inhibited,especially in group MC2w,the proliferation of lymphocytes and monocytes infiltration in cardiac grafts.There was no significant difference in serum IL-10 and Foxp3 mRNA and protein in cardiac grafts and spleen of recipients among the groups (P＞0.05).Conclusion Induction of CM in recipients could relieve cardiac allograft rejection and prolong cardiac allograft survival via its anti-inflammation and anti-apoptotic effects,not via up-regulation of Foxp3 in recipient mice.

Objective To evaluate the feasibility and efficacy of catheter-directed thrombolysis (CDT) for the treatment of non-acute (history ＞ 14 days) deep venous thrombosis (DVT) of the lower extremity.Methods Clinical data of 63 patients of non-acute DVT of lower extremities treated by CDT and adjunctive angioplasty and stenting from July 2009 to August 2013 were analyzed retrospectively.Venous recanalization was graded by a thrombus score based on pre-and post-treatment venography.Follow-up was performed by Doppler ultrasound and clinical evaluation.Results A total of 63 limbs with DVT with a mean symptom duration of (22 ± 5) days were treated by a continuous combined with pulse-spray infusion of urokinase of (1.21 ± 0.69) million IU/d for (74 ± 21) hours.Significant recanalization was achieved in 77％ (48 of 63) of the treated limbs.After thrombolysis,percutaneous angioplasty was done for 15 residual lesions and stent placement was performed in 11 iliac veins and 1 femoral vein.Minor bleeding occurred in 6 (10％) patients,no patients suffered from major bleeding or symptomatic pulmonary embolism.During follow up (mean:15 ±6 months),the veins were patent in 45 (71％) limbs.15 (24％) limbs developed mild post-thrombotic syndrome (PTS),and none had severe PTS.Conclusions CDT combined with adjunctive angioplasty and stenting is safe and effective for removal of the clot burden and for restoration of the venous flow in patients with non-acute lower extremity DVT.

Objective To investigate the consistency of M RI detecting posterior ligamentous complex (PLC) injury associated with thoracolumbar factures.Methods MRI data of 170 cases of thoracolumbar fractures were reviewed retrospectively.Each case underwent MRI around one week postinjury.MRI data were analyzed and compared by three physicians respectively to discuss the consistency in MRI detection of PLC injury and the severity of PLC injury.Results Kappa coefficient was 0.846 between observer 1 and 2,0.768 between observer 1 and 3,and 0.793 between observer 2 and 3.Interobserver reliability was high and overall Kappa coefficient was 0.803.Severity of PLC injury was interrelated with spinal cord nerve injury (P ＜ 0.05).Conclusions Accurate detection of PLC injury in thoracolumbar fractures is beneficial to clear the mechanical stability of the spine.MRI detection of PLC injury is of high consistency and hence deserves wide use.

【Objective】To observe the dynamic interference of cellular endotoxin specific receptors of Toll-like receptor 4(TLR4) mRNA and nuclear factor ?B p65(NF-?B p65) in rats with damp-heat syndrome of seasonal febrile disease by Ganlu Xiaodu Dan(GXD),and to explore the therapeutic mechanism of prescriptions with the actions of clearing heat and resolving dampness.【Methods】Wistar rats were randomized into 3 groups: A(normal control),B(models established by feeding of high-fat diet and colon bacillus under the high-temperature and high-humidity condition),and C(rat models treated with GXD).Reverse transcription-polymerase chain reaction(RT-PCR) analysis was used to detect the TLR4 mRNA level in hepatic macrophages and immunohistochemical method was used to detect the activation of NF-?B p65.【Results】The expression of TLR4 mRNA and NF-?B p65 was increased gradually in group B 6,12 and 24 hours after infection with colon bacillus,and the differences were significant,indicating that the expression of TLR4 mRNA and NF-?B p65 increased with the prolongation of disease course.In group C,TLR4 mRNA expression was decreased in different time points after infection,the difference of TLR4 mRNA expression 12 and 24 hours after infection was significant as compared with group B and group A(P

Objective To observe the protective effects of salidroside on glutamate-induced injury in cultured hippocampal neurons.Methods Primarily cultured hippocampal neurons from fetal Wistar rat were incubated with salidroside(10,20 and 40mg/L) for 24 hours,then glutamate(125?mol/L) was added for 15 minutes to induce injury.Cell viability was detected by MTT assay,the vigor of LDH was determined by biochemistry method,the apoptosis rates were anallyzed using Annexin V-FITC and PI labelling and Hoechst 33342 staining and flow cylometric assay.Fluorescent intensity of intracellular free calcium was observed with laser scanning confocal microscopy(LSCM).Results After the pretreatment with salidroside for 24 hours,the increases of LDH vigor and apoptosis rates and the decrease of cell viability caused by glutamate were resisted obviously.Salidroside inhibited the increase of Ca~2+ in cytoplasm significantly.Conclusion Salidroside can significantly resist the injury induced by glutamate.The neuroprotective activities of salidroside can be related to its ability to reduce Ca~2+ overload in cytoplasm.

The aim of this study was to investigate the effect of Compound Yihe Tea on improving insulin resistance in obesity mice. Thirty-two male C57BL/6J mice were randomly divided into 4 groups: the normal fat diet group(NFD group), high fat diet group(HFD group), Compound Yihe Tea low dosage group［20 mg/(kg ·d), YH-L group］ and high dosage group［40 mg/(kg ·d), YH-H group］. NFD group was given standard feed, and the remaining mice were administered with high fat diet. After 6 weeks, YH-H and YH-L groups were given Compound Yihe Tea for 6 weeks. Blood glucose was measured at week 11 and serum levels of total cholesterol(TC), serum triglyceride(TG), low-density lipoprotein cholesterol(LDL-C)and high-density lipoprotein cholesterol(HDL-C)were measured at week 12. Liver tissues were prepared for oil red O and HE staining. Immunohistochemical analysis was used to test the protein expression of GLUT4 in liver. Protein expressions of PI3K, Akt and GLUT4 in epididymis white adipose tissue(WAT)were tested by Western blot. The results showed that Compound Yihe Tea could effectively reduce body weights and the serum levels of TC, TG and LDL-C. Furthermore Compound Yihe Tea could improve the histopathological changes of liver, up-regulate the protein expression of PI3K, Akt and GLUT4 in epididymis WAT and the protein expression of GLUT4 in liver. Compound Yihe Tea can reduce the fat accumulation in liver tissue, improve the indexes of blood glucose and lipid levels, and improve insulin resistance via PI3K-AKT-GLUT4 pathway.

Objective To investigate the damage sequence of posterior ligamentous complex (PLC) and its clinical significance in thoracolumbar fracture.Methods Data of 132 patients with spinal fracture evaluated with X-rays,CT and short-tau inversion-recovery (STIR) sequences in MRI were collected prospectively.Fracture morphology was classified using the AO classification.PLC components including interspinous ligament (ISL),supraspinous ligament (SSL),ligamentum flavum (LF) and facet capsules (FC) were assessed and classified as intact,edema,or tear.ISL edema was further subdivided depending on the extension (＞ 50％ or ≤50％).Correlation between MRI signal and AO progressive scale of morphological damage was analyzed.Results AO type A1/A2 fracture associated with only FC distraction.AO type A3 fracture showed additional ISL tear,usually less than 50％,with neither LF nor SSL tear.AO type B1 fracture showed FC distraction,ISL edema or disruption,and low rate of SSL/LF tear,but B2 fracture increased the rate of SSL/LF tear.AO type C fracture showed facet fracture or dislocation and ISL,SSL as well as LF tear.High correlation was found between AO progressive scale and MRI signal (P ＜ 0.01).Conclusions MRI study can well display the PLC damage and damage sequence.MRI correlates with AO progressive scale of morphological damage,which shows a progressive orderly rupture sequence among different PLC components as traumatic forces increase.

Objective To knockout and identify the Antigen 43 (Ag43) in the Escherichia Coli JM109 .Methods Mutation group Ⅱ introns RNA protein complexes (RNP) gene sequence was obtained by Sigma Company′s TargeTron Gene Knockout Sys‐tem and Ag43 gene specific designed PCR primers amplification ,then ,to acquired Ag43 specific recombinant RNP plasmid pACD4K‐Ag4 ,this gene sequence was inserted into the plasmid pACD4K‐C of RNA′s expression .Finally ,pEGFP‐Ag43 was trans‐formed into JM109 and inserted the group Ⅱ intron into the Ag43′s locus by IPTG inducing expression .Results The best insertion locus was between 1 812 and 1 913 .Through the agarose electrophoresis gel ,the RNP gene sequence was consistent with the expec‐ted value (350 bp) .The pEGFP‐Ag43 vector was correctly constructed which was proofed by endonuclease Nhe Ⅰ and Hind ⅡI di‐gestion as predicted products (3 646 and 4 029 bp;7 000 and 550 bp ,respectively ) .The PCR and gene sequence results indicated that the group Ⅱ intron was inserted into the locus between 1 812 and 1 913 in the Ag43 gene .Conclusion Successful knockout of the Ag43 in Escherichia Coli JM109 found basis to further study the Ag43′s function and regard the coli as host bacteria of Ag43 chimeric protein recombinant .

Objective To observe the effect of bone marrow-derived mesenchymal stem cells (bMSCs) on graft healing within a bone tunnel after anterior cruciate ligament (ACL) reconstruction in rabbits.Methods The study involved 24 New Zealand white rabbits undergone ACL reconstruction with an autologous ipsilateral gastrocnemius tendon graft.Both hindlimbs were included.In one hindlimb,graft coated with fibrin glue compound by bMSCs was employed (bMSCs group).Whereas in the contralateral hindlimb,graft coated with fibrin glue without cells was employed (control group).At postoperative 2,4,6 and 8 weeks,specimens were harvested to have a biomechanical test of tensile strength and stiffness of tendon-bone interface.Results Tensile strength and stiffness of tendon-bone interface in both experiment and control groups presented a rising trend with the prolong of repair time.In contrast,significantly higher tensile strength and stiffness of tendon-bone interface were observed in experiment group since the 6 weeks (P ＜ 0.05).Conclusion bMSCs transplantation significantly enhances the early tensile strength and stiffness at tendon-bone interface after ACL reconstruction in rabbits and improves the graft healing within a bone tunnel.

Objective To understand the knowledge of tuberculosis in general hospital patients with respiratory diseases.Methods Face to face questionnaire survey 821 respiratory patients, the main contents include: the tuberculosis awareness, common symptoms of tuberculosis, tuberculosis with or without contagious, persistent cough expectorant willing to go to where the treatment of tuberculosis, national policy, treatment of the course of treatment, tuberculosis prevention measures, and whether the knowledge of tuberculosis drug resistance.Results 92.1% of patients were aware of TB and 98.7% of patients knew TB was contagious, and 99% knew that TB was transmitted by the respiratory tract.Only 4.5% of patients with symptoms appear willing to go to tuberculosis treatment of patients.94.9% of the patients were aware of a specific TB control facility, 85% knew that the basic TB treatment was free, 99% thought to be good treatment of tuberculosis patients, 55% of patients know that tuberculosis treatment for a long time, to regular medication.30% of people know that mycobacterium tuberculosis resistance.90% of patients know to isolate tuberculosis patients, 40% of patients that protect susceptible populations, and 95% believed that BCG could prevent tuberculosis.Conclusion Patients have a certain understanding of tuberculosis, the timely treatment of indifference to the treatment of treatment is probably not impressed, very little knowledge of tuberculosis resistance.