Objective: To explore research the protective effect of ischemia preconditioning to rabbits′myocardium injury of ischemia reperfusion, and discuss its mechanism. Methods: We randomly divide rabbits into 3 groups ( 8 rabbits every group ) , namely non-operation group . ischemia reperfusion group ( IR ) . ischemia preadaptation group ( IP ), and observed the level of enzymatic dynamics of myocardium,SOD,MDA, ET and correlation peptide of calcitonin gene. Results: In terms of the level of enzymatic dynamics of myocardium, SOD,MDA, ET and CGRP, the differences between IR and IP are very obviously (P

Objective To analyze the urodynamic data of the patients with benign prostatic hyperplasia(BPH)and provide evidence for the therapy.Methods 452 patients with BPH were selected.The mean age was(65.6 ±5.65)years(51 to 89 years).All patients had intact history documents which included the results of cystoscopy,urodynamics,doppler,DRE and PSA.Results 15 cases were not diagnosed with BOO.430 cases were diagnosed with BOO,among them,65 cases of bladder detrusor muscle weakness,bladder detrusor instability(detrusor,DI,instability)in 365 cases,3 cases were normal,and the other 4 cases had detrusor atony.In 445 patients the mean maximum urinary flow rate(8.32±3.15)ml/s,the average residual urine volume(87.68±79.46)ml,the average maximum flow rate when the detrusor pressure(62.32±7.54)cm H2O,maximum urethral closure pressure was(86.43±18.35)cmH2O,among which 19 cases were not suitable for surgery.At the end of the 426 patients underwent surgical treatment,postoperative follow-up of 3 to 18 months,the International Prostate Symptom Score(IPSS)from 0 to 5,the quality of life index(QOL)score of 0 to 1,Qmax 11 to 21 ml /s,residual urine(45.6±36.2)ml.There were significant differences between the preoperative and postoperative indexes(P<0.05).Conclusion Urodynamic tests can avoid the blindness in the surgery treatment of BPH.

Objective:To investigate the inhibitory effect and mechanism of different doses of Xiao Chai Hu Tang on C6 glioma cells cultured in vitro. Methods:C6 glioma cells were inoculated in 96 holes,24 holes and 6 holes,each culture plate was divided into 4 groups:control group, low dose group, middle dose group and high dose group, when the cells covered the bottom of culture plate 80%-90%,began adding,cultured for 24 hours after the ter mination of training. Cell proliferation activity,cell viability,protein content and protein positive expression intensity of VEGF and ESM-1,cell apoptosis in early and late stage were detected by CCK-8,in vivo staining,ELISA, immunocytochemistry and flow cytometry. Results: CCK-8 assay showed that the growth of C6 glioma cells was inhibited by low,medium and high dose group;ELISA and immunocytochemistry showed that the expression of VEGF and ESM-1 was lower in the lower, middle and high dose groups, and the levels of protein expression and protein levels were decreased. The flow cytometry showed that the low dose of small radix,middle and high dose group could promote the cell apoptosis. Inverted microscope ob-servation showed that with the increase of dose,the number of cells increased gradually,and the number of dead cells increased,and all kinds of detection methods showed that the inhibition effect increased with dose and dose dependence. The difference between the two groups was statistically significant(P<0. 01). Conclusion:The growth of C6 glioma cells was significantly inhibited by Xiao Chai Hu Tang. It may play a role in inhibiting tumor growth by down regulating ESM-1 and VEGF protein level and promoting cell apoptosis.

Objective To investigate the biomechanicai characteristics of flexor profundus tendons repaired after decimeter wave therapy, and to observe the effect of decimeter wave therapy on early active mobilization. Methods A total of 56 Leghorn chickens were randomly divided into a therapy group and a control group with 28 chickens in each. The 3rd and 4th toes of their left feet were employed for the establishment of a tendon injury model. The flexor profundus tendons were cut and repaired. Gypsum support was applied and fixed with an adhesive plaster after the operation. The operated sites on toes Ⅲ and Ⅳ were exposed. The external fixation was removed 3 weeks later and the chickens were left free to move. Decimeter wave therapy ( frequency 915 MHz, power 8 Watts) was ap-plied for 10 minutes once daily on the left foot of each chicken in the therapy group from day 1 until 3 weeks after the operation. Sham decimeter wave therapy was applied to chickens in the control group. Four chickens from each group were randomly selected at the 1st, 7th, 10th, 14th, 18th, 21st and 28th days for biomechanical analysis. Biome-chanical parameters including tensile strength of rupture (Pmax), elongation ratio at rupture (δimax) and the tensile adhesion strength of the rupture zone (W0>) were observed at each time point. Results At the 7th, 10th, 14th, 18th, 21st and 28th day after the operation, the differences in Pmax, δmax and W0 between the therapy and control groups were statistically significant. The results of the therapy group were better than those of the control group. Conclusions Local decimeter wave therapy after flexor tendon repair can promote intrinsic healing and reduce ex-trinsic healing. The speed and quality of healing are improved. The elasticity and tenacity of the injured tendons are enhanced. Therefore decimeter wave therapy is helpful for early active mobilization training.

OBJECTIVETo explore the effects of stress imposed on adenoid cystic carcinoma (ACC), therefore to clarify the molecular basis and mechanism of ACC's malignant phenotype under the elevated tumor interstitial fluid pressure.

METHODSACC cells were cultured under pressure (103.74 kPa), and were divided into four groups (3 h group, 6 h group, 12 h group, 24 h group) according the pressure time. Untreated ACC2 was as negative control group, untreated ACCM was as positive control group. The level of epidermal growth factor receptor (EGFR) was detected by semiquantitative analysis of immunochemistry. Matrix metalloproteinase 9 (MMP9) and EGFR mRNA expression were assessed by reverse transcriptase polymerase chain reaction. EGFR, phosphorylation epidermal growth factor receptor (P-EGFR), MMP9, keratinocyte growth factor (KGF) and phosphorylation extracellular signal-regulated kinase (P-ERK) protein expressions were assessed by Western blot.

RESULTSAs the extension of pressure time, the expression of EGFR, P-EGFR, MMP9, KGF, P-ERK in ACC2 gradually increased, which were positive correlation with pressure time, and were higher than that of negative control group.

CONCLUSIONUnder the stimulation of pressure, the mRNA and protein levels of adhesion molecules and metastatic relative molecules in ACC2 were sharply elevated.

Objective:Cancer stem cells (CSCs) are resistant to chemotherapy. Our study aimed to investigate the stem cell-like proper-ties of doxorubicin-resistant osteosarcoma cell line 143B and its correlation with Notch signaling. Methods:We generated doxorubicin-resistant osteosarcoma cells by treating them with 2μm doxorubicin. Stem cell-like properties such as morphology change, Stro-1/CD117 double positive ratio, stem cell-related gene expression, sphere formation efficiency, and EMT character were assessed on day 5 after doxorubicin withdrawal. Notch receptor and its target genes were examined using qPCR and Western blot analysis. The stem cell-like properties of doxorubicin-resistant osteosarcoma cells were assessed when pretreated with Notch inhibitor or vehicle. The an-ti-tumor effect of Notch inhibitor was tested using a xenograft model. Results:Doxorubicin-resistant osteosarcoma cells were enriched in Stro-1+/CD117+cells, which showed obvious increased expression of stem cell-related genes, and exhibited enhanced spheroid for-mation and evident mesenchymal characteristics unlike doxorubicin-sensitive cells. qPCR and Western blot assays showed that Notch intracellular domain 1 (NICD1) and target genes Hes1 and Hey1 were upregulated in doxorubicin-resistant osteosarcoma stem cells compared with those in vehicle cells. Furthermore, pretreatment with a γ-secretase inhibitor (GSI) to prevent Notch signaling en-hanced chemo-sensitivity and inhibited doxorubicin-enriched osteosarcoma stem cell activity in vitro. Finally, the Notch inhibitor pre-vented tumor growth in mice xenograft models. Conclusion: Doxorubicin induced the enrichment of osteosarcoma stem-like cells through Notch signaling, and inactivation of Notch could be useful for overcoming drug resistance and eliminating osteosarcoma.

In order to develop a safer vaccine strain exploit Salmonella Pullorum vaccine strain as a live vaccine vector.Methods:AΔcrpΔasd mutant of S.pullorum C79-13 strain was constructed and it was developed E.coli donor strain mutant was generated through the two-step method introduced by χ7213 ( pREΔasd) was conjugated with the recipient of C 79-13Δcrp.The C79-13ΔcrpΔasd the transduction of recombinant suicide plasmid .Results:PCR and sequencing results showed that ΔsipBSL1344 was suc-cessfully constructed.The further study indicated that foreign DAP must be supplied for the ΔcrpΔasd mutant to grow,in addition,the asd gene was transmitted stably .Compared with C79-13 strain,the O antigens was identical to C79-13 strain,but the growth velocity was reduced significantly ,significantly reduced virulence .Conclusion: The ΔcrpΔasd mutant can accept asd+plasmid to construct host-vector balance lethal system , and this system can be used to express foreign gene efficiently and to develop potential oral multivalent vaccines.

Objective: To develop an oral live vaccine vector which stably carries exogenous genes.Methods:SL1344ΔsipBΔasd host-vector balanced lethal system was constructed by the method of recombinant suicide plasmid-mediated allelic exchange on the basis of attenuated Salmonella typhinurium SL1344ΔsipB.Then,the biological characteristics of SL1344ΔsipBΔasd was analyzed.Results:The results showed that the mutant was stabile with the Δasd gene in vitro;the serotype and growth rate of SL1344ΔsipBΔasd strain was almost same as the parent SL1344ΔsipB and SL1344 strain.And the mutant strains remain swim ming zones.Virulence test in mice showed that the virulence of SL1344ΔsipBΔasd which carried complementary plasmid pYA3493 by electro-transformation decreased by 1.4%compared with SL1344.Conclusion: These results showed that the SL1344ΔsipBΔasd mutant was successfully constructed.It is likely that this mutant should be used as a live vector to express foreign genes.

Radiotherapy is a treatment for cancer with undesired by-effects. In order to develop a new radiation protective agent that could reduce the by-effects, we tried to express and purify human cryptochrome 1 (hCRY1). The coding sequence of hCRY1 was inserted into prokaryotic expression plasmid pET28a(+), and this protein was purified from Escherichia coli BL21(DE3) after IPTG induction, ultrasonication, inclusion body dissolution, gradient dialysis, nickel column purification and ultrafiltration. The yield of hCRY1 in 1 L E. coli culture (LB medium) was about 10-15 mg. The radiation protective efficiency of hCRY1 was monitored by detecting X-ray-induced H2A.X foci in HaCaT cells. The results of immunofluorescence show that hCRY1 significantly reduces X-ray stimulated DNA damage response. The apoptosis of HaCaT cell was also detected, and the repression of H2A.X foci formation was not due to hCRY1's cytotoxity. All these data suggest a potential application of recombinant hCRY1 as a protective agent for radiotherapy.
Cryptochromes ; biosynthesis ; Escherichia coli ; Humans ; Plasmids ; Radiation-Protective Agents ; Recombinant Proteins ; biosynthesis

Objective:In order to develop a safer vaccine strain exploit Salmonella typhimurium vaccine strain .A ΔsipB mutant of Salmonella typhimurium SL 1344 strain was constructed.Methods: Firstly, the recombinant suicide plasmid containing the missing 585 bp sipB ( PREΔsipB ) was built by homologous recombination , and screenned by two-step method.Results: PCR and sequencing results showed that SL 1344ΔsipB was successfully constructed.It was no significant changes compared with SL 1344.But compared with the parent strains SL 1344 , the mutant strain had obvious change in its virulence , oral challenge of bacteria in mice revealed that LD50 of the mutant strain was 1.70 ×108 CFU,the toxicity reduced about 1.4%.The protection rate induced by the sipB mutant was 50%,and the serum antibody peaked 14 d post-immunization.Conclusion:The SL1344ΔsipB mutant was constructed suc-cessfully,and genetic stability ,significantly reduced virulence.The study provides a new approach for further study of the relationship between the gene and pathogenicity of Salmonella typhimurium.It is likely that the ΔsipB mutant could be adapted to develope attenuated Salmonella vaccine.