Objective To construct a technical platform for scarless gene modification of Yersinia pestis and to study the functions of its specific genes.Methods The resistance fragment, including upstream and downstream homologous arms of targeted regions, was reamplified by asymmetric PCR.The amplicons were introduced into Y.pestis harboring plasmid pKD46.With the induction of L-arabinose,the recombinant related enzymes: Exo, Beta and Gam, were expressed to guide the homologous recombination.A donor plasmid, pKSI-1, which carried the desired modification fragment flanking by I-SceⅠ recognition sites, was introduced into Y.pestis as the second step of λ-Red recombination with the help of pREDTKI.Results and Conclusion Two mutant strains:△waaA and waaA(△9nt), were successfully constructed for Y.pestis strain 201.Scarless modification introduces no extra modification to the genome, and it is ideal for comprehensive functional genomic studies.

Objective To investigate the roles of SENP1 in regulation of biological characteristics of NK cells.Methods Lentivirus-mediated-Senp1-small-hairpinRNA (shRNA) transduction was applied to NK92 cells.The expression of SENP1 in NK92 cells was determined by real-time PCR and Western blot.The proliferation of NK92 cells was detected by CCK-8 assay.The apoptosis of NK92 cells was determined by Annexin Ⅴ and PI labeling.The cytotoxicity of NK92 cells against K562 cells was evaluated by luciferase reporter assay.Results Treatment of NK92 cells with IL-21 resulted in SENP1 upregulation.Lentivirus mediated SENP1 knockdown reduced proliferation and increased apoptosis in NK-92 cells,but SENP1 inhibition had slight impact on the cytotoxic ability of NK92 cells to kill K562 cells.Conclusion SENP1 mediates the regulatory effect of IL-21 on the proliferation and survival of NK92 cells.

Objective To investigate the influence of dexmedetomidine by continuous intravenous infu-sion on the recovery process of patients undergoing laparoscopic radical hysterectomy surgery under general anes-thesia.Methods Eighty patients,ASAⅠ~Ⅱ,scheduled for laparoscopic radical hysterectomy surgery under general anesthesia,were randomized into group C and group D.Normal saline and dexmedetomidine at the dose of 0.6μg.kg-1 .h-1 were injected into different groups from 10 mins before the operation to 20 mins before the end of operation respectively.Heart rate(HR),mean arterial pressure(MAP) were recorded at following five time points:before anesthesia(T1),10 mins before extubation(T2),extubation(T3),5 mins after extubation(T4), 10 mins after exbubation(T5).The recovery time,cough reflex score,sedation-agitation scale(SAS),Ramsay score,the occurrence rate of untoward effect and the dosage of medication were observed.The observational indi-ces were analyzed by using t-test,chi-square test,repeated measures data of ANOVA or the Mann-Whitney u test method.Results Compared with group C,during the recovery process,MAP and HR in group D was more stable.Ramsay score in group D was higher(P<0.05).The SAS in group D was lower(P<0.05).The sedation-agitation scale in group D was lower(P<0.05).The occurrence rate of agitation and tachycardia was lower in group D(P<0.05).But the occurrence rate of bradycardia was higher(P<0.05).Meanwhile,usage amount of sevoflurane was lower in group D(P<0.05).Conclusion Dexmedetomidine continuous intravenous infusion re-duced the untoward effect of extubation,did not extend extubation time,and kept more stable haemodynamics.

Bacteriophages are abundantly distributed viruses , which are able to infect bacteria .Bacteriophages are becoming a focus of attention for microbiologists , as they can cause pollution to the fermentation industry and might serve as alternative therapies for antibiotic-resistant pathogenic bacteria .Effective bacteriophage infection normally involves adsorp-tion, injection, replication, assembly and release , against which bacteria have developed various resistance strategies .The research progress in the bacteriophage resistance mechanisms is briefly reviewed in this paper .

Objective To evaluate the therapeutic effect of hepatocyte growth factor(HGF) gene modified placenta-derived mesenchymal stem cells( PMSCs) on limb ischemia in a rabbit model.Methods The placental tissue was digested with enzyme, cultured and passaged.The PMSCs were characterized by surface marker expression.These cells were infected with adenoviral( Ad)-HGF and intramuscular injected for treatment of limb ischemia in a rabbit model.The blood supply of the limb was detected by digital subtraction angiography ( DSA ) and the vessel number was evaluated in histopathological HE staining.Results The results showed that Ad-HGF gene transduction increased the vascular endothelial growth factor ( VEGF ) , basic fibroblast growth factor, bFGF ( bFGF ) and HGF expression in PMSCs. Transplantation of HGF-transduced PMSCs resulted in the increase in vessel density and improvement of blood supply in the rabbit limb ischemia model.Conclusion The therapeutic effect of HGF gene engineered PMSCs on ischemia by enhancing angiogenesis in a rabbit model is evaluated.Transplantation of PMSCs with HGF gene therapy may be a promising strategy for the treatment of ischemia diseases.

Objective To investigate the prevalence rates of healthcare-associated infection (HAI)and antimicrobial use in a hospital in 2011-2013,and provide scientific basis for prevention and control of HAI.Methods HAI prevalence rates and antimicrobial use in a hospital in 2011-2013 were investigated by combination of bedside visiting and medical records reviewing.Results A total of 3 011 patients were investigated during three years,the prevalence rates were 3.49%, 2.87% and 3.98% respectively,difference of prevalence rates during three years were not significantly different (χ2 =2.105,P =0.356).Among HAI sites,lower respiratory tract ranked first,the major pathogens were gram-negative bacil-li,constituent ratios of different pathogens were not significantly different among three years(χ2 =1.003,P =0.972);anti-microbial usage rates and specimen detection rates among 3 years were both significantly different(χ2 =12.569,P <0.01;χ2=6.758,P <0.01,respectively),antimicrobial usage rate was highest in 2011(63.40%),and specimen detection rate was highest in 2012(62.14%).Conclusion Point prevalence rate in this hospital is at average national level,antimicrobial usage rate decreased, the consciousness of pathogenic detection gradually enhanced,clinical application management of antimicrobial agents still needs to be strengthened continuously.

BACKGROUNDSleep deprivation (SD) has been used in treatment of depression disorder, and could effectively improve the patients' depressive symptoms.The aim of the study was to explore the effects of SD on electroencephalographic (EEG) and executive function changes in patients with depression.

METHODSEighteen depression patients (DPs) and 21 healthy controls (HCs) were enrolled in the present study. The whole night polysomnography (PSG) was recorded by Neurofax-1518K (Nihon Kohden, Japan) system before and after 36 hours of SD. The level of subjects' depression state was assessed by Visual Analogue Scale (VAS), and the executive function was assessed by Wisconsin Card Sorting Test (WCST).

RESULTSSignificantly decreased sleep latency (SL; before SD: (31.8 ± 11.1) minutes, after SD: (8.8 ± 5.2) minutes, P < 0.01) and REM sleep latency (RL; before SD: (79.8 ± 13.5) minutes, after SD: (62.9 ± 10.2) minutes, P < 0.01) were found after SD PSG in depression patients. Decreased Stage 1 (S1; before SD: (11.7 ± 2.9)%, after SD: (7.3 ± 1.1)%, P < 0.01) and Stage 2 (S2, before SD: (53.8 ± 15.5)%, after SD: (42.3 ± 14.7)%, P < 0.05) of non-rapid eye movement (NREM) sleep, and increased Stage 3 (S3, before SD: (11.8 ± 5.5)%, after SD: (23.6 ± 5.8)%, P < 0.01) and Stage 4 (S4, before SD: (8.8 ± 3.3)%, after SD: (27.4 ± 4.8)%, P < 0.01) NREM sleep were also found. After SD, the depression level in patients decreased from 6.7 ± 2.1 to 2.9 ± 0.7 (P < 0.01). In WCST, the patients showed significantly decreased Response errors (Re, before SD: 22.3 ± 2.4, after SD: 18.3 ± 2.7, P < 0.01) and Response preservative errors (Rpe, before SD: 11.6 ± 3.6, after SD: 9.3 ± 2.9, P < 0.05). Depression patients' RE (t = 2.17, P < 0.05) and Rpe (t = 2.96, P < 0.01) also decreased significantly compared to healthy controls.

CONCLUSIONSD can improve depression symptom and executive function in depression patients.

Adult ; Depression ; physiopathology ; Female ; Humans ; Male ; Middle Aged ; Polysomnography ; methods ; Sleep Deprivation ; physiopathology

Objective To construct a prostate cancer specific oncolytic adenovirus armed with a fusion protein gene , PSA-IZ-CD40L, and to evaluate its oncolytic efficiency and immune activation ability in vitro.Methods Prostate Specific Antigen (PSA) gene, CD40L-N and CD40L-C genes were obtained from cDNA of LNCaP cells and Jurkat cells using poly-merase chain reaction (PCR) or nested-PCR, respectively.PSA,Linker,CD40L-N and CD40L-C were linked sequentially to generate fusion protein gene PSA-IZ-CD40L (PL) by overlapping PCR.Then, prostate specific oncolytic adenovirus PL-carrying gene, Ad-PL-PPT-E1A,was constructed using the oncolytic adenovirus system , which was based on Adeasy sys-tem.PC3M cells were infected by Ad-PL-PPT-E1A at serial multiplicity of infection (MOI), and the apoptosis was detec-ted by flow cytometry at several time points post-infection.For immune activation detection , PC3M cells were infected with Ad-PL-PPT-E1A at a MOI of 50, and the cell lysate was collected at 48 h post-infection.Peripheral blood mononuclear cells derived (PBMCs) from healthy donors were stimulated by the lysate from PC 3M cells or Ad-PL-PPT-E1A infected PC3M cells before proliferation was assayed using cell counting kit-8 (CCK8).Results Fusion protein gene, PSA-IZ-CD40L, was successfully constructed and cloned into the prostate cancer specific adenovirus to generate Ad -PL-PPT-PL. The expression of E1A and PL protein could be detected by reverse transcription PCR and Western-blotting.Cytopathic effect was observed in PC3M cells infected with Ad-PL-PPT-E1A.Furthermore, the apoptosis rate reached 70.67% ± 2.98%at 48 h post-infection with 200 MOI Ad-PL-PPT-E1A.Compared with the lysate of PC3M cells, that from Ad-PL-PPT-E1A infected cells could promote the proliferation of PBMCs .Conclusion We have constructed a prostate cancer spe-cific oncolytic adenovirus armed can fusion protein gene PL , Ad-PL-PPT-E1A, which could kill PC3M cells effectively and enhance the proliferation of PBMCs in vitro.

Objective To study the effect of heavy ion radiation on proliferation and apoptosis of human peripheral blood derived T lymphocytes and the mechanism .Methods T lymphocytes were isolated from heparinized whole blood samples by density gradient centrifugation using Ficoll before being irradiated with heavy ion beams 12 C.The accumulated absorbed dose (dose-rate values=0.5 Gy/min, and meanLET=29 keV/μm).12 h and 24 h post-infection, total RNA of T lymphocytes was isolated , and the apoptosis related gene expression , including Bcl-2, Bax, Caspase3, Caspase8 and Caspase9, was detected by RT-RT-PCR.24 h and 48 h after irradiation, the proliferation was analyzed by CCK 8 kit.The cell apoptosis was detected by flow cytometry after being labeled with AnnexinV-PE/7-AAD or AnnexinV-FITC/PE.The expression of Bcl-2, Bax and Caspase3 was also assayed by RT-PCR.Results Data showed that heavy ion radiation could inhibit the proliferation of T lymphocytes obviously , and the inhibition ratio in cells that received 2 Gy dose was much high-er than in cells that received 1 Gy dose.Furthermore, heavy ion radiation promoted the apoptosis of T lymphocytes signifi-cantly.The results of RT-PCR showed that the mRNA expression of Bcl-2 was down-regulated in heavy ion radiation T lym-phocytes while the expression of Bax and Caspase 3 was up-regulated.Conclusion Heavy ion radiation can inhibit the pro-liferation and promote the apoptosis of human peripheral blood derived T lymphocytes .

In this paper, we present a new method which can reconstruct the three-dimensional model of nasolacrimal duct. We firstly resampled the volume data along nasolacrimal duct direction, then segmented the nasolacrimal duct into slices, and finally, completed the 3D reconstruction from the two-D contours. Using this method, we can not only reconstruct normal nasolacrimal duct, but also reconstruct catagmatic nasolacrimal duct. It overcomes the current shortcomings of some traditional methods. Consequently, the technology proposed is of great significance in computer aided technique and surgical planning related to ophthalmonogy.
Humans ; Image Processing, Computer-Assisted ; methods ; Imaging, Three-Dimensional ; methods ; Nasolacrimal Duct ; anatomy & histology ; diagnostic imaging ; surgery ; Reconstructive Surgical Procedures ; methods ; Tomography, X-Ray Computed