Objective To investigate the effects of Sulfotanshinone Sodium Injection (SSI) on neuropathic pain in rats.Methods One hundred and eight adult male Wistar rats,aged 6-8 weeks,weighing 180-220 g,were randomly divided into 3 groups (n =36 each):sham operation group (group S) ; chronic constrictive injury (CCI)group; group SSI.The animals were anesthetized with intraperitoneal 10% chloral hydrate 350 mg/kg.In groups CCI and SSI,4 ligatures were placed on the right sciatic nerve at 1 mm intervals with 4-0 silk thread according to the method described by Bennett et al.In group S,the right sciatic nerves were exposed,but not ligated.In group SSI,SSI 25 mg/kg was injected intraperitoneally once a day starting from the end of operation until one day before the animals were sacrificed,while the rats received the equal volume of normal saline (5 ml/kg) instead of SSI in groups S and CCI.Twelve animals in each group were chosen at 1 day before operation and 3,7 and 14 days after CCI (T1-4) to measure mechanical paw withdrawal threshold to yon Frey stimuli (MWT) and paw withdrawal latency to thermal nociceptive stimulus (TWL).Six rats in each group were sacrificed at T2-4 after measurement of pain threshold,and their lumbar segnents (L4-6) of the spinal cord were immediately removed for determination of Bcl2 and caspase-3 expression in spinal dorsal horn (by immune-histochemistry),and MDA content and SOD activity (by spectrophotometry) in spinal cord.Results Compared with group S,PWT was significantly decreased,PWL was shortened,the expression of Bcl-2 and caspase-3 was up-regulated,MDA content was increased and SOD activity was decreased at T2-4 in groups CCI and SSI (P ＜ 0.05).Compared with group CNP,PWT was significantly increased,PWL was prolonged,the expression of Bcl-2 was up-regulated,the expression of caspase-3 was downregulated,MDA content was decreased and SOD activity was increased at T2-4 in group SSI (P ＜ 0.05).Conclusion SSI can mitigate neuropathic pain in rats and inhibition of oxidative stress in spinal cord tissues and reduction of apoptosis in spinal dorsal horn neurons are involved in the mechanism.

Objective To evaluate the effects of dexmedetomidine on the activity of cAMP response element binding protein (CREB) and c-fos in the spinal dorsal horn in a rat model of neuropathic pain.Methods Fifty-four adult male Wistar rats,aged 6-8 weeks,weighing 180-220 g,were randomly divided into 3 groups (n =18 each):sham operation group (group S),chronic neuropathic pain group (group C) and dexmedetomidine group (group D).The animals were anesthetized with intraperitoneal 10％ chloral hydrate 350 mg/kg.The sciatic nerve was exposed and 4 ligatures were placed on the right sciatic nerve at 1 mm intervals with 4-0 silk thread in C and D groups.In group D,dexmedetomidine 50 μg/kg was injected intraperitoneally once a day starting from the end of operation until 1 day before the animals were sacrificed,while the equal volme of normal saline was injected instead of dexmedetomidine in S and C groups.Paw withdrawal threshold to mechanical stimulation with yon Frey filament (MWT) and paw withdrawal latency to thermal stimulation (TWL) were measured on 1 day before operation and 3,7 and 14 days after operation.The animals were sacrificed after measurement of MWT and TWL.Their lumbar segments (L4-6) of the spinal cord were removed for measurement of the expression of phosphorylated CREB (pCREB) and c-fos by immunohistochemistry.Results Compared with group S,MWT was significantly decreased,TWL was shortened,and the expression of pCREB and c-fos was up-regulated on 3,7 and 14 days after operation in C and D groups (P ＜ 0.05).Compared with group C,MWT was significantly increased,TWL was prolonged,and the expression of pCREB and c-fos was down-regulated on 3,7 and 14 days after operation in group D (P ＜ 0.05).MWT was significantly lower,and TWL was shorter on 3,7 and 14 days after operation than on 1 day before operation in C and D groups (P ＜ 0.05).MWT was significantly lower,TWL was shorter,and the expression of pCREB and c-fos was higher on 7 and 14 days after operation than on 3 days after operation in C and D groups (P ＜ 0.05).MWT was significantly higher,TWL was longer,and the expression of pCREB and c-fos was lower on 14 days after operation than on 7 days after operation in C and D groups (P ＜ 0.05).Conclusion The mechanism by which dexmedetomidine reduces neuropathic pain is related to inhibition of the activity of CREB and c-fos in the spinal dorsal horn of rats.

Objective To evaluate the effect of dexmedetomidine and small dose of ketamine on the expression of P2X4 receptor (P2X4 R) mRNA and P2X7 receptor (P2X7R) mRNA in the dorsal root ganglion of rats with neuropathic pain.Methods Ninety male Sprague-Dawley rats,aged 6-9 weeks,weighing 180-220 g,were randomly divided into 5 groups (n =18 each):sham group (group S),chronic constrictive injury group (group CCI),dexmedetomidine group (group D),ketamine group (group K) and dexmedetomidine + ketamine group (group DK).The animals were anesthetized with intraperitoneal 10％ chloral hydrate 400 mg/kg.Neuropathic pain was induced by CCI in CCI,D,K and DK groups.The sciatic nerve was exposed and 4 loose ligatures were placed on the sciatic nerve at 1mmintervals with 4-0 silk thread.In group S,the sciatic nerves were only exposed but not ligated.In D,K and DK groups,dexmedetomidine 50μg/kg,ketamine 10 mg/kg and dexmedetomidine 25μg/kg + ketamine 5 mg/kg were injected intraperitoneally,respectively,while the equal volume of normal saline was injected in S and CCI groups,once a day for 14 consecutive days after CCI.Mechanical paw withdrawal threshold (MWT) and thermal paw withdrawal latency (TWL) were measured at 1 day before CCI,and 3,7 and 14 days after CCI.Six animals were sacrificed after measurement of pain threshold at 3,7 and 14 days after CCI and the lumbar segments (L4-6) of the dorsal root ganglion were removed for determination of P2X4 R mRNA and P2X7 R mRNA expression by RT-PCR.Results Compared with group S,MWT and TWL were significantly decreased at 3,7 and 14 days after CCI in groups CCI,D,K and DK,the expression of P2X4R mRNA and P2X7R mRNA was up-regulated at 3,7 and 14 days after CCI in groups CCI,D and K,and the expression of P2X4 R mRNA and P2X7 R mRNA was up-regulated at 3 and 7 days after CCI in group DK (P ＜ 0.05).Compared with group CCI,TWL and MWT were significantly increased and the expression of P2X4 R mRNA and P2X7 R mRNA was down-regulated at 3,7 and 14 days after CCI in groups D,K and DK (P ＜ 0.05).Compared with D and K groups,TWL and MWT were significantly increased and the expression of P2X4 R mRNA and P2X7 R mRNA was down-regulated at 3,7 and 14 days after CCI in group DK (P ＜ 0.05).Conclusion The mechanism by which the combination of dexmedetomidine and small dose of ketamine produces a synergistic antinociception in rats with neuropathic pain may be related to down-regulation of the expression of P2X4 R mRNA and P2X7 R mRNA.

Objective To evaluate the effect of dexmedetornidine on the expression of Toll-like receptor 4 (TLR4) and nuclear factor kappa B (NF-κB) in the spinal cord in rats with neuropathic pain (NP).Methods One hundred and eight male Wistar rats,aged 6-8 weeks,weighing 180-220 g,were randomly assigned into 3 groups (n =36 each):sham operation group (group S),NP group and dexmedetomidine group (group D).NP was induced by chronic constrictive injury in anesthetized rats.Sciatic nerve was exposed and 4 loose ligatures were placed on the sciatic nerve at 1 mm intervals with 4-0 silk thread.In group S,the right sciatic nerves were exposed,but not ligated.Dexmedetomidine 50 μg/kg was injected intraperitoneally once a day from the onset of operation to one day before the rats were sacrificed in group D,while the equal volume of normal saline was injected in groups S and NP.Mechanical withdrawal threshold (MWT) and thermal pain threshold (TPT) were measured on the day before operation (T0) and 3,7,and 14 days after operation (T1-3).After measurement of pain threshold at T1,T2 and T3 after operation,the L4-6 segments of the spinal cord were removed for determination of the expres-sion of TLR4 and NF-κB mRNA (by RT-PCR) and the expression of TLR4 and NF-κB in spinal dorsal horn (by immuno-histochemistry).Results Compared with group S,MWT and TPT were significantly decreased and the expression of TLR4,NF-κB and TLR4 and NF-κB mRNA was up-regulated after operation in groups NP and D (P ＜ 0.05).Compared with group NP,TPT and MWT were significantly increased and the expression of TLR4,NF-κB,TLR4 mRNA and NF-κB mRNA was significantly down-regulated after operation in group D (P ＜ 0.05).Conclusion The mechanism by which dexmedetomidine attenuates NP in rats is related to inhibition of the expression of TLR4 and NF-κB in rat spinal cord.

OBJECTIVE To investigate the pathogenic causes of lower respiratory tract infections(LRTIs) in the elderly in Guangzhou area.METHODS Pathogens obtained from 107 patients with LRTIs were performed by multiple diagnostic tools that including bacterial culture,PCR and specific immunological assays.RESULTS A bacterial cause was established in 42(68.5%) and an atypical pathogen cause in 25(31.6%) of the 107 patients.Mycoplasma pneumoniae and Haemophilus influenzae remained the most important pathogens for LRTIs.CONCLUSIONS In the prescription of antibiotics in the elderly with LRTIs,not only bacteria but also atypical pathogens should be taken into account.

PEG-modified magnetic Fe3O4 (Fe3O4-PEG) nanoparticles were sythesized using a solvothermal reaction and characterized with transmission electron microscopy (TEM) and thermo gravimetric analysis (TGA). The photothermal effect and photothermal destruction of cancer cells were evaluated. Then the doxorubicin loaded Fe3O4-PEG (DOX-Fe3O4-PEG) nanoparticles were prepared. The cytotoxicity and combined chemotherapy/photothermal therapy (PTT) effect were investigated. Uniform PEG coated Fe3O4 nanoparticles with particle size of 155 nm were obtained in the experiment. The loading and release of doxorubicin on Fe3O4-PEG were pH-dependent. The drug loading capacity in water was 21%. The results of MTT indicated a good biocompatiblity of Fe3O4-PEG nanoparticles and high cytotoxicity of DOX-Fe3O4-PEG. In combined therapy experiment, photothermal therapy demonstrated unambiguously enhanced chemotherapy efficacy. In conclusion, the obtained Fe3O4-PEG nanoparticles which exhibit good photothermal effect and drug loading capacity can be used for chemotherapy and photothermal therapy. The synergetic anti-tumor activity indicates the potential for the combined application of chemotherapy and photothermal therapy in cancer treatment.
Antibiotics, Antineoplastic ; administration & dosage ; pharmacology ; Cell Survival ; drug effects ; Doxorubicin ; administration & dosage ; pharmacology ; Drug Carriers ; Ferrosoferric Oxide ; chemistry ; Humans ; Hyperthermia, Induced ; MCF-7 Cells ; Magnetite Nanoparticles ; chemistry ; Particle Size ; Polyethylene Glycols ; chemistry

Objective To investigate prevalence and risk factors of urgency urinary incontinence (UUI) among adult women in Beijing.Methods Multiple-stage stratified sampling was used to recruit women equal to and more than 20 years old from 48 communities ( villages and neighborhoods ) in six districts of Beijing,by household interview with a standardized questionnaire to collect information of demographic characteristics,delivery history,health conditions and UUI related clinical data.Data analysis was performed by SPSS software version 16.0.Results Totally,3058 adult women completed the survey,with an overall prevalence of UUI of 1.7 % (52/3058).Distribution of age,cultural background,family income,number of pregnancy,number of abortion,delivery history,history of chronic diseases,general surgery,pelvic surgery,abdominal pain and abdominal distension were found significantly different between UUI participants and non-UUI ones with univariate analysis ( all P ＜ 0.05 ).Results of multivariate logistic regress analysis showed that number of pregnancy ( OR = 1.29,95% CI = 1.06-1.58 ),history of chronic diseases ( OR = 2.53,95% CI = 1.38-4.62 ) and history of abdominal pain ( OR = 2.32,95%CI= 1.10 -4.91 ) all significantly associated with UUI.Conclusions UUI associates with complicated factors and its prevention and intervention at communities is necessary targeting at related factors.

PEG-modified magnetic Fe3O4 (Fe3O4-PEG) nanoparticles were sythesized using a solvothermal reaction and characterized with transmission electron microscopy (TEM) and thermo gravimetric analysis (TGA). The photothermal effect and photothermal destruction of cancer cells were evaluated. Then the doxorubicin loaded Fe3O4-PEG (DOX-Fe3O4-PEG) nanoparticles were prepared. The cytotoxicity and combined chemotherapy/photothermal therapy (PTT) effect were investigated. Uniform PEG coated Fe3O4 nanoparticles with particle size of 155 nm were obtained in the experiment. The loading and release of doxorubicin on Fe3O4-PEG were pH-dependent. The drug loading capacity in water was 21%. The results of MTT indicated a good biocompatiblity of Fe3O4-PEG nanoparticles and high cytotoxicity of DOX-Fe3O4-PEG. In combined therapy experiment, photothermal therapy demonstrated unambiguously enhanced chemotherapy efficacy. In conclusion, the obtained Fe3O4-PEG nanoparticles which exhibit good photothermal effect and drug loading capacity can be used for chemotherapy and photothermal therapy. The synergetic anti-tumor activity indicates the potential for the combined application of chemotherapy and photothermal therapy in cancer treatment.

OBJECTIVETo establish a human embryonic stem cell line with stably β-catenin gene silencing by lentivirus-mediated shRNA interference.

METHODSPLKO.1-puro-β-catenin vector, a lentivirus plasmid expressing β-catenin shRNA, was packaged into 293FT cells. Human embryonic stem cells were infected with the lentivirus and the cell clones stably expressing β-catenin shRNA were selected by puromycin, with the uninfected cells and cells infected with the empty vector as the control. Real-time RT-PCR was used to evaluate the efficiency of β-catenin knocked down; β-catenin and OCT4 protein expression in the infected cells was examined using immunofluorescence assay.

RESULTSInfection with β-catenin-specific shRNA lentivirus resulted in stable interference of β-catenin expression in human embryonic stem cells, which showed a β-catenin mRNA expression of only 1% of that in the uninfected cells. Infection with the empty vector produced no obvious effect on β-catenin expression compared to the uninfected cells. In the cells infected with β-catenin shRNA lentivirus, β-catenin protein expression was almost undetectable in immunofluorescence assay, while OCT4 was still expressed after the interference.

CONCLUSIONLentiviral vector-delivered shRNA results in effective and stable β-catenin gene silencing in human embryonic stem cells.

Cell Line ; Embryonic Stem Cells ; cytology ; metabolism ; Genetic Vectors ; Humans ; Lentivirus ; genetics ; Plasmids ; RNA Interference ; RNA, Small Interfering ; genetics ; Transfection ; Wnt Signaling Pathway ; beta Catenin ; genetics ; metabolism

Objective Through experiments in vitro,we explored the role of Chinese medicine monomer-Astragaloside Ⅳ with DC vaccine in the body immunologic function.Methods Peripheral blood mononuclear cells (PBMCs)from healthy volunteers were isolated,cultured and generated in vitro,pulsed with tumor antigen from SGC7901 gastric carcinoma cell lysates,produced DC vaccine.Observe T-cell proliferation responses stimulated by DC vaccine with AⅣ group,DC vaccine group and AⅣ group respectively,and the anti-tumor effects on SGC7901 cells in vitro.Results ①The T-cell proliferation rate of DC vaccine with AⅣ group and DC vaccine group were significantly higher than AⅣ group and T-cell group(negative control group)(P=0.000).The stimulating efficacy on T-cell proliferation of DC vaccine with AⅣ group was higher than that of DC vaccine group(S/R 1∶5,1∶10,1∶50,1∶100,P=0.013,0.014,0.017,0.019).Compared with T-cell group,the T-cell proliferation rate of AⅣ group had no statistically significance(P=0.185).②The killing rate of effector cells actived by DC vaccine with AⅣ group and DC vaccine group against SGC7901 gastric carcinoma cells were higher than that of AⅣ group and T-cell group(P=0.000).The Killing power of DC vaccine with AⅣ group was stronger than that of DC vaccine(E/T 5∶1,10∶1,20∶1,50∶1,P=0.023,0.012,0.016,0.011);while the group of AⅣ group and T-cell group cannot killing tumor cells.Both had no statistically significance(P=0.267).Conclusion AⅣ can stimulate T-cell proliferation and enhance the activity of killing tumor cells by DC,which induced specific antitumor response against stomach carcinoma cells effectively.