OBJECTIVE To investigate antibiotic resistance in nosocomial infections with Acinetobacter baumannii from lower respiratory tract in the intensive care unit(ICU) and guide clinically rational use of drug.METHODS Antibiotics susceptibility tests in 109 clinical isolates of A.baumannii from lower respiratiory tract were performed by K-B disk diffusion method.The results were judged according to CLSI 2007.As controls,standard strain was used simultaneously.RESULTS The sensitivity rate to cefoperazone/sulbactam was the highest,arriving at 77.1% and followed by imipenem(57.8%) and meropenem(54.9%).The other antibiotics were below 50.0%,generally.The serious cross-resistance phenomenon was in present.Pan-resistant strains had been detected.CONCLUSIONS The drug-resistant status in nosocomial infections with A.baumannii from lower respiratory tract in ICU is very serious.We should continue to strengthen the monitoring of the antibiotic resistance and prevent the outbreak epidemics of drug resistant strains in ICU.

Background:Traditional tissue engineering methods to fabricate urinary tract patch have some drawbacks such as compromised cell viability and uneven cell distribution within scaffold.In this study,we combined three-dimensional (3D) bioprinting and tissue engineering method to form a tissue-engineered urinary tract patch,which could be employed for the application on Beagles urinary tract defect mode to verify its effectiveness on urinary tract reconstruction.Methods:Human adipose-derived stem cells (hADSCs) were dropped into smooth muscle differentiation medium to generate induced microtissues (ID-MTs),flow cytometry was utilized to detect the positive percentage for CD44,CD105,CD45,and CD34 of hADSCs.Expression of vascular endothelial growth factor A (VEGFA) and tumor necrosis factor-stimulated gene-6 (TSG-6) in hADSCs and MTs were identified by Western blotting.Then the ID-MTs were employed for 3D bioprinting.The bioprinted structure was encapsulated by transplantation into the subcutaneous tissue of nude mice for 1 week.After retrieval of the encapsulated structure,hematoxylin and eosin and Masson's trichrome staining were performed to demonstrate the morphology and reveal collagen and smooth muscle fibers,integral optical density (IOD) and area of interest were calculated for further semiquantitative analysis.Immunofluorescent double staining of CD31 and a-smooth muscle actin (α-SMA) were used to reveal vascularization of the encapsulated structure.Immunohistochemistry was performed to evaluate the expression of interleukin-2 (IL-2),α-SMA,and smoothelin of the MTs in the implanted structure.Afterward,the encapsulated structure was seeded with human urothelial cells.Immunofluorescent staining of cytokeradns AE1/AE3 was applied to inspect the morphology of seeded encapsulated structure.Results:The semi-quantitative assay showed that the relative protein expression of VEGFA was 0.355 ± 0.038 in the hADSCs vs.0.649 ± 0.150 in the MTs (t--3.291,P =0.030),while TSG-6 expression was 0.492 ± 0.092 in the hADSCs vs.1.256 ± 0.401 in the MTs (t =3.216,P =0.032).The semi-quantitative analysis showed that the mean IOD of IL-2 in the MT group was 7.67 ± 1.26,while 12.6 ± 4.79 in the hADSCs group,but semi-quantitative analysis showed that there was no statistical significance in the difference between the two groups (t =1.724,P =0.16).The semi-quantitative analysis showed that IOD was 71.7 ± 14.2 in noninduced MTs (NI-MTs) vs.35.7 ± 11.4 in ID-MTs for collagen fibers (t =3.428,P =0.027) and 12.8 ± 1.9 in NI-MTs vs.30.6 ± 8.9 in ID-MTs for smooth muscle fibers (t=3.369,P=0.028);furthermore,the mean IOD was 0.0613 ±0.0172 in ID-MTs vs.0.0017 ± 0.0009 in NI-MTs for α-SMA (t =5.994,P =0.027),while 0.0355 ± 0.0128 in ID-MTs vs.0.0035 ± 0.0022 in NI-MTs for smoothelin (t=4.268,P =0.013),which indicate that 3D bioprinted structure containing ID-MTs could mimic the smooth muscle layer of native urinary tract.After encapsulation of the urinary tract patch for additional cell adhesion,urothelial cells were seeded onto the encapsulated structures,and a monolayer urothelial cell was observed.Conclusion:Through 3D bioprinting and tissue engineering methods,we provided a promising way to fabricate tissue-engineered urinary tract patch for further investigation.

Immunohistochemistry and double immunofluorescent labeling techniques combined with confocal laser scanning microscope analysis were used to investigate the characteristic spatial induction profile of nestin following a transient middle cerebral artery occlusion in adult rat brain. The results showed that nestin was induced in ischemic core at 1 day after reperfusion. In addition to ischemic core, the expression of nestin increased in peri-ischemic I, II and III regions at 3 days and 1 week, then it decreased and narrowed along the rim of ischemic core 2 weeks after reperfusion. Double immunofluorescent labeling showed that nestin positive cells were mostly co-stained with GFAP,a astrocyte marker, in peri-ischemic I region 3 days after reperfusion. At 2 weeks, however nestin cells showed a long process and the cells double stained with nestin and NSE,a neuonal specific marker,increased in the ischemic brain. The results suggest that cerebral ischemia induces nestin expression in damaged neurons which might favor the neuroprotection against ischemic damage.
Animals ; Brain ; metabolism ; pathology ; Brain Ischemia ; metabolism ; pathology ; Immunohistochemistry ; Infarction, Middle Cerebral Artery ; metabolism ; pathology ; Nestin ; metabolism ; Neurons ; metabolism ; Rats

OBJECTIVETo investigate the surgical treatment of benign prostate hyperplasia (BPH) and its concomitant diseases at the same time.

METHODSOne hundred and fourteen operations were performed for BPH patients, including transurethral resection/vapor of the prostate (TURP/TUVP), inguinal herniorrhaphy, internal urethrotomy, transurethral resection of bladder tumor (TURBt) or vesical litholapaxy, and the data were reviewed.

RESULTSThe procedures were successful in all cases. A follow-up of 3 to 60 months found a good outcome of TURP. There was no recurrence in 30 cases of inguinal hernia and 39 cases of vesical calculus. Of the 25 cases of urethral stricture, 1 had an obvious hypotension during the operation and 4 needed urethral dilatation after operation. Six of the 20 cases of bladder tumor underwent a second TURBt due to the recurring tumor which was far from prostatic urethra.

CONCLUSIONInguinal hernia, urethral stricture, bladder tumor or vesical calculus can be treated simultaneously during TURP.

Aged ; Aged, 80 and over ; Follow-Up Studies ; Hernia, Inguinal ; complications ; surgery ; Humans ; Male ; Middle Aged ; Prostatic Hyperplasia ; complications ; surgery ; Transurethral Resection of Prostate ; Urethral Stricture ; complications ; surgery ; Urinary Bladder Calculi ; complications ; surgery ; Urinary Bladder Neoplasms ; complications ; surgery

BACKGROUNDH3N2 subtype influenza A viruses have been identified in humans worldwide, raising concerns about their pandemic potential and prompting the development of candidate vaccines to protect humans against this subtype of influenza A virus. The aim of this study was to establish a system for rescuing of a cold-adapted high-yielding H3N2 subtype human influenza virus by reverse genetics.

METHODSIn order to generate better and safer vaccine candidate viruses, a cold-adapted high yielding reassortant H3N2 influenza A virus was genetically constructed by reverse genetics and was designated as rgAA-H3N2. The rgAA-H3N2 virus contained HA and NA genes from an epidemic strain A/Wisconsin/67/2005 (H3N2) in a background of internal genes derived from the master donor viruses (MDV), cold-adapted (ca), temperature sensitive (ts), live attenuated influenza virus strain A/Ann Arbor/6/60 (MDV-A).

RESULTSIn this presentation, the virus HA titer of rgAA-H3N2 in the allantoic fluid from infected embryonated eggs was as high as 1:1024. A fluorescent focus assay (FFU) was performed 24-36 hours post-infection using a specific antibody and bright staining was used for determining the virus titer. The allantoic fluid containing the recovered influenza virus was analyzed in a hemagglutination inhibition (HI) test and the specific inhibition was found.

CONCLUSIONThe results mentioned above demonstrated that cold-adapted, attenuated reassortant H3N2 subtype influenza A virus was successfully generated, which laid a good foundation for the further related research.

Animals ; COS Cells ; Cercopithecus aethiops ; Dogs ; Hemagglutinin Glycoproteins, Influenza Virus ; genetics ; Influenza A Virus, H3N2 Subtype ; immunology ; Influenza Vaccines ; immunology ; Mice ; Mice, Inbred BALB C ; Neuraminidase ; genetics ; Plasmids ; Reassortant Viruses ; immunology ; Reverse Transcriptase Polymerase Chain Reaction ; Vaccines, Attenuated ; immunology ; Viral Proteins ; genetics

OBJECTIVEA lot of studies have suggested that a certain amount of T cells may be involved among cytokine-induced killer (CIK) cells. The aim of the present study was to prove whether an antigen-specific killing effect on tumor cells is involved during the CIKs-induced killing process.

METHODSBone marrow mononuclear cells (BMMNCs) derived from healthy subjects were separately cultured to generate dendritic cells (DC) and CIKs. A human mammary cancer cell line MCF-7/ADR, expressing P-gp antigen, was frozen-thawed and the lysate including P-gp antigen was obtained. The DC pulsed with or without tumor antigen lysate was co-cultured with CIK (pulsed-DC + CIK and DC + CIK), and CIK cultured alone was used as control. The cell phenotype of DC and CIK was analyzed by flow cytometry. The secretion of IL-12 and IFN-gamma was assayed by ELSA. The antitumor effect of the three CIK groups targeted at MCF-7/ADR cells expressing P-gp antigen and MCF-7 cells was detected by MTT.

RESULTSPulsed-DC + CIK group and DC + CIK group showed a higher expression level of DC mature phenotypes than those before co-culture with CIK, with a significant difference (P = 0.003, P = 0.001, respectively). The phenotypes (CD3, CD8, CD56) of CIK in pulsed-DC + CIK group and DC + CIK group was higher than those in CIK group (P = 0.003, P = 0.011, respectively). Among the three CIK groups, pulsed-DC + CIK group had the highest phenotypes on CD3+ CD56 (pulsed-DC + CIK vs. DC + CIK, P = 0.001; pulsed-DC + CIK vs. CIK, P < 0.001) and CD3 CD8 (P = 0.002, P = 0.002, respectively). Among the three groups, the pulsed-DC + CIK group showed the lowest CD45RA phenotype (pulsed-DC + CIK vs. DC + CIK, P < 0.001; pulsed-DC + CIK vs. CIK, P = 0.004). Among the three groups the secretion of IL-12 and IFN-gamma had the highest level in pulsed-DC + CIK group, with a value of 254 +/- 14.5 pg/ml and 3100 +/- 286 pg/ml, respectively. The antitumor killing effect on MCF-7/ADR cells had a significant difference between any two groups (pulsed-DC + CIK VS. DC + CIK, P = 0.039; pulsed-DC + CIK VS. CIK, P = 0.002; DC + CIK vs. CIK, P = 0.049). The highest was in pulsed-DC + CIK group and the lowest was in CIK group. The CIK group showed a significantly lower antitumor effect on MCF-7 cells than the other two groups (pulsed-DC + CIK vs. CIK, P = 0.007; DC + CIK vs. CIK, P = 0.048), but no significant difference between the pulsed-DC + CIK and DC + CIK groups.

CONCLUSIONIn the present study, DC and CIK cells have been successfully obtained and cultured from bone marrow mononuclear cells. After their co-culture, not only both their specific phenotypes were increased, but also the associated cytokines were secreted. An improved antitumor killing effect and some possible specific immunocytotoxicity were observed. Our findings provided a basis for experimental and clinical research on bio-immunotherapy targeted at multi-drug resistant tumor cells.

ATP-Binding Cassette, Sub-Family B, Member 1 ; metabolism ; Breast Neoplasms ; metabolism ; pathology ; CD3 Complex ; metabolism ; CD56 Antigen ; metabolism ; CD8 Antigens ; metabolism ; Cell Line, Tumor ; Cell Proliferation ; Coculture Techniques ; Cytokine-Induced Killer Cells ; cytology ; immunology ; metabolism ; Cytotoxicity, Immunologic ; Dendritic Cells ; cytology ; immunology ; metabolism ; Drug Resistance, Multiple ; Drug Resistance, Neoplasm ; Female ; Humans ; Interferon-gamma ; metabolism ; Interleukin-12 ; metabolism

This paper reviewed the present nursing manager succession planning at home and abroad, covering the succession standard of nursing managers, the content and form of pre-job training, and the evaluation tool of management ability. These efforts aim to provide references for the construction of nursing management talents teams in China.

Objective:To establish a HPLC method for determination of chlorogenic acid,rutin,polydatin,kaempferol-3-O-rutinoside,astragalin,resveratrol,quercetin,kaempferol in Tetrastigmatis Hemsleyani Radix,in order to study the content changes of eight components of different months. Method:Zorbax SB C₁₈column (4.6 mm×250 mm,5 μm)was adopted with mobile phase consisting of acetonitrile(A)and 0.1%phosphoric acid(B)in gradient elution(0-30 min,10%-30%A;30-40 min,30%-95%A;40-45 min,95%A;45-60 min,95%-10%A). The flow rate was 0.8 mL·min^-1,the column was kept at 25℃,and the detection wavelength was 320 nm. Result:Chlorogenic acid,rutin,polydatin,kaempferol-3-O-rutinoside,astragalin,resveratrol,quercetin,kaempferol showed a good linearity within the range of 13.7-549 mg·L^-1(r=0.999 0),12.6-253 mg·L^-1(r=0.999 1),15.8-316 mg·L^-1(r=0.999 0),14.7-147 mg·L^-1(r=0.999 2),8.8-88 mg·L^-1(r=0.999 1),7.9-79 mg·L^-1(r=0.999 5),8.6-172 mg·L^-1(r=0.999 1),8.9-89 mg·L^-1(r=0.999 4). There were great differences in contents of the eight flavonoid active components in different growth phases. In July and August,the relative contents of chlorogenic acid,rutin,kaempferol 3-O-rutinoside and polydatin were the highest. The highest relative content of quercetin was observed in June. The relative contents of resveratrol and kaempferol in April and May was higher than those in other mouths. The relative content of astragalin in November was the highest. Conclusion:It could provide abundant information for the production and quality control of Tetrastigmatis Hemsleyani Radix.

BACKGROUND: Preparing a scaffold with cartilage derived components and good initial mechanical strength is the direction of tissue engineering cartilage research. OBJECTIVE: To prepare porous acellular osteochondral scaffolds, and to explore their mechanical properties and cell compatibility. METHODS: Osteochondral bone from the porcine knee joint was taken, and then porous osteochondral scaffolds were made by laser microporation technology. Subsequently, the scaffolds were decellularized chemical methods. Scaffold structure was observed by scanning electron microscopy, and the compression modulus of the scaffolds was determined. Bone marrow mesenchymal stem cells were cultured in L-DMEM containing 10% fetal bovine serum (control group) and cultured in the medium extract of porous acellular osteochondral scaffolds (experimental group), respectively. Cell proliferation was detected by cell counting kit-8 method within 5 days of culture. Bone marrow mesenchymal stem cells were seeded on the porous acellular osteochondral scaffolds, and within 28 days of co-culture, cell growth was observed by hematoxylin-eosin staining and toluidine blue staining. RESULTS AND CONCLUSION: (1) Observation under scanning electron microscopy: The porous acellular osteochondral scaffolds had the smooth surface with evenly distributed pores. The pores of the scaffold extended longitudinally into the subchondral bone. (2) Mechanical properties: The average compressive modulus of porous acellular osteochondral scaffolds was 0.77 MPa, which was close to the compression modulus of the normal cartilage (1.15 MPa). (3) Cell counting kit-8 test: There were no differences in cell proliferation between the control and experimental groups at 1, 2, 3, 4 and 5 days of culture. (4) Cell-scaffold co-culture: A large amount of bone marrow mesenchymal stem cells were observed to be adherent to the scaffold after 1 day of culture through hematoxylin-eosin and toluidine blue staining. However, as time went on, a few cells adhered to the pore wall or grew into the pores at 7 and 21 days of culture. There were also some adherent cells but a large amount of cell masses formed in the pores at 28 days of culture. To conclude, the porous acellular osteochondral scaffold has good mechanical properties and cell compatibility.

Objective To investigate the seasonal fluctuations of mosquitoes in Taibai Lake area of Jining City,and evaluate the application effects of two mosquito trapping tools(Gongfu Xiaoshuai mosquito lamp and WJ-C mosquito sampler),so as to provide references for the effective surveillance,prevention and control of local mosquito vectors.Methods The overnight cap-ture of mosquitoes with a man-bait net trap,Gongfu Xiaoshuai mosquito lamp,and WJ-C mosquito sampler was performed to col-lect mosquitoes.The species of adult mosquitoes and seasonal fluctuations were investigated and analyzed.Results A total of 10 914 mosquitoes belonging to 13 species,5 genera were collected with the overnight capture of mosquitoes with the man-bait net trap,and Culex tritaeniorhynchus,Cx.pipiens palllen and Mansonia uniformis were the dominant species,accounting for 77.41％of the total.The proportion of Coquillettidia ochracea increased significantly.All the adult mosquitoes appeared in June,and reached the density peak in July and August.Both of the two surveillance tools showed relatively sensitive capture perfor-mance against mosquitoes.The trapping results of WJ-C mosquito sampler were basically the same as the man-bait trapping meth-od,however,the amount of Mansonia uniformis and Coquillettidia ochracea collected by Gongfu Xiaoshuai mosquito lamp was significantly lower than that of the man-bait trapping method.Conclusions The mosquito species are varied in Taibai Lake area of Jining City.Therefore,monitoring should be strengthened.In addition,the two mosquito trapping tools are sensitive,but they have both advantages and disadvantages,thus we should better pay attention to the consistency of monitoring methods in use.