Protective effects of ulinastatin on cardiac dysfunction in mice with heat stroke and its mechanism / 解放军医学杂志

Objective To examine the effects ofulinastatin (UTI) on cardiac dysfunction in mice with heat stroke and its possible mechanism.Methods 20 mice were divided into four groups randomly:room temperature plus normal saline (Sham+NS),room temperature plus UTI (Sham+UTI),heat stress plus normal saline (HS+NS),heat stress plus UTI (HS+UTI),5 each.105U/kg UTI was delivered by intraperitoneal injection before the onset of the heat stress.Room temperature groups were housed at room temperature (23.0 ± 0.5 ℃),while heat stress groups were kept in an incubator at 36.5 ± 0.5 ℃ and humidity of 65.0％ ± 2.0％.The rectal temperature (Tr) reaching 42℃ was taken as severe heat stroke,and the time in two heat stress groups was recorded.The mice were transferred to the room temperature (23.0 ± 0.5 ℃) for natural cooling after the heat stroke onset.6 hours after the treatment,cardiac output (CO) was ultrasonographically detected,the myocardium was separated for histopathological examination and the expression of total p38 and phosphorylated p38 (p-p38) was determined by Western blotting.Results The time to reach 42℃ in HS+UTI group was significantly prolonged (P=0.044).Compared with the Sham+NS group,the CO in HS+NS and HS+UTI group decreased significantly (P=0.017),and the score of myocardial inflammation (P＜0.001) and p-p38/p38 ratio (P＜0.001) increased.The CO was significantly higher in HS+UTI group than in HS+NS group (P=0.030),and the score of myocardial inflammation (P＜0.001) and p-p38/p38 ratio (P=0.001) were significantly lower.Conclusion Ulinastatin might improve the cardiac function in mice with heat stroke by decreasing the p-p38 and alleviating the inflammation response.

Puerarin alleviates HUVEC injury induced by visfatin through up-regulating miR-155-3p / 中国实验动物学报

Object To study the effect of puerarin on the expression of inflammatory factors and miR-155-3p in human umbilical vein endothelial cells ( HUVEC) induced by visfatin.Methods The HUVEC cell injury model was es-tablished with visfatin.Cell proliferation was measured by MTT assay.Cell apoptosis was detected by flow cytometry.The level of CRP and NF-κB was detected by ELISA, and the expression of miR-155-3p was detected by RT-PCR.The expres-sion of myeloid differentiation factor 88 ( MyD88) was identified by western blotting.Results Visfatin induced cell prolif-eration and inhibited apoptosis in HUVEC, meanwhile the expressions of both CRP and NF-κB were significantly increased, compared with that of the control group (P<0.01).Puerarin at moderate and high concentrations obviously reduced the HUVEC injury induced by visfatin, mainly through down-regulating the expression of CRP and NF-κB, as well as up-regu-lating the level of miR-155-3p in the HUVEC.MiR-155-3p mimic markedly decreased the level of MyD88, CRP and NF-κB in the HUVEC induced by visfatin (P<0.05).Conclusions Pueprarin obviously alleviates HUVEC injury induced by visfatin, probably related to down-regulating the level of MyD88, CRP, NF-κB, and up-regulating the expression of miR-155-3p in HUVEC.

Prevalence of Hearing Disorders Based Whole Population in Jilin Province, China / 中国康复理论与实践

Objective To investigate the prevalence, etiology, rehabilitation demands and service condition of hearing disorders based on the whole population in Jilin Province, China. Methods Using the probability proportion to size (PPS) sampling, 9246 (93.3%) out of 9909 residents sampled form 36 counties were targeted for investigation from August, 2014 to January, 2015, followed the WHO Ear and Hearing Disorders Survey Protocol. The hearing loss and disability were classified as WHO recommended and Classification and Grading Criteria of Disability (GB/T 26341-2010). Results The standardized prevalence of hearing loss and disability was 16.41%and 4.78%, re-spectively. Age, sex, residence, occupation and marriage status, education level and household income were significantly associated with hearing loss prevalence, while nationality was not. The main etiologies included non-infectious disease (47.33%), ear disease (14.17%), un-known causation (13.89%), and noise (8.59%). Among all people with hearing loss, those who accepted intervention service accounted for 11.02%. Among all people with hearing disability, those who used hearing aids accounted for 5.58%, and 0.67%used artificial cochlea. Con-clusion Demographics and socioeconomic factors are significantly associated with the prevalence of hearing loss. The main etiology con-tains non-infectious disease, ear disease and noise. Both the rate of service utilization among people with hearing loss and the rate of adopt-ing hearing aids among people with hearing disability are low. It is needed to do more in prevention and rehabilitation of hearing impairment.

The study of external factors affecting auditory ability of infants and toddlers after cochlear implantation / 临床耳鼻咽喉头颈外科杂志

OBJECTIVE@#To analyze external factors affecting auditory ability of infants and toddlers after cochlear implantation in the first year of switch-on.@*METHOD@#Seventy-five infants and toddlers after cochlear implantation were selected as subjects, using LittlEARS Auditory Questionnaire to assess and analyze the correlations with auditory ability and external factors (including gender, cochlear implanted age, pre-implant hearing aid fitting, caregivers' education background, household income and rehabilitation modes) in different stages (before switch-on, and 3 months, 6 months, 9 months, 12 months after switch-on).@*RESULT@#The mean scores of LittlEARS were significantly different in cochlear implanted age group, pre-implant hearing aid fitting group and rehabilitation modes group (P 0.05). The correlations with the mean scores of LittlEARS and cochlear implantation age or pre-implant hearing aid fitting were significant at 3 months or 6 months after switch-on(/r/ ≥ 0. 3, P ≥ 0.3, P < 0.01).@*CONCLUSION@#Cochlear implanted age and pre-implant hearing aid fitting were the important factors affecting auditory ability of infants and toddlers after cochlear implantation in the first year of switch-on. The effect of rehabilitation modes on auditory ability of infants and toddlers after cochlear implantation was slow.

Mechanism Study of Xiaoyao San in Improving Depression-Like Behavior of Rats by Regulation of Hepatic Sinusoidal Endothelial Function / 广州中医药大学学报

Objective To observe the effectiveness and mechanism of Xiaoyao San (Xiaoyao Powder for Soothing Liver and Relieving Depression) in improving depression-like behavior of rats. Methods Male Wistar rats were randomized into normal group, model group, Xiaoyao San (1.9 g·kg-1·d-1) group, and fluoxetine (2 mg·kg-1·d-1) group. The rats were exposed to chronic unpredictable mild stress ( CUMS) to induce rat depression-like behavior. Field test was performed for the observation of effect of Xiaoyao San on rat depression-like behavior, Luminex liquid chip system was applied to detect the serum cytokines, and the amount and size of rat hepatic sinusoidal endothelial window were examined under electron microscope, and hepatic indoleamine 2, 3-dioxygenase ( IDO) and tryptophan 2, 3 -dioxygenaes ( TDO) expression levels were detected by immunohistochemical and Western blot methods. Results Xiaoyao San showed obvious effect on increasing sugar water consumption, the number of crossing the blocks and erection frequency in rats, decreasing serum levels of tumor necrosis factor alpha ( TNF-α) and interleukin 6 ( IL-6) , increasing the amount of hepatic sinusoidal endothelial window, promoting hepatic sinusoidal endothelial vascularization, and reducing TDO and IDO expression ( P<0.05 or P<0.01). Conclusion Xiaoyao San exerts obvious effect on improving rat depression-like behaviors, and the mechanism is probably related with the decrease of inflammatory factors, inhibition of IDO pathway, and improvement of hepatic sinusoidal endothelial function.

Amentoflavone induces apoptosis in SW480 human colorectal cancer cells via regulating β-catenin and caspase-3 expressions / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the role of β-catenin and caspase-3 in amentoflavone-induced apoptosis of human colorectal cancer SW480 cells.</p><p><b>METHODS</b>MTT assay was used to detect the viability of SW480 cells exposed to amentoflavone, and flow cytometry was employed to assess the cell apoptosis. Western blotting was performed to determine the protein expressions of β-catenin and caspase-3 in the exposed cells.</p><p><b>RESULTS</b>Amentoflavone dose-dependently inhibited the viability of SW480 cells, and a high concentration of amentoflavone (150 µmol/L) obviously induced apoptosis of the cells. Amentoflavone exposure caused significantly increased expression of caspase-3 and suppressed β-catenin expression in the cells.</p><p><b>CONCLUSION</b>Amentoflavone-induced apoptosis in SW480 human colorectal cancer cells is associated with altered expressions of β-catenin and caspase-3.</p>

Lentivirus-mediated LOX-1-RNA interference attenuates oxidative stress-induced apoptosis in myocardial cells / 南方医科大学学报

<p><b>OBJECTIVE</b>To construct a lectin-like oxidized low-density lipoprotein receptor (LOX-1) RNA interference (RNAi) lentivirus and explore the role of LOX-1 in H(2)O(2)-induced apoptosis of rat myocardial cells.</p><p><b>METHODS</b>LOX-1 shRNA sequence was synthesized and cloned into pLentiLox3.7 (pLL3.7) lentiviral vector to construct the lentiviral vector pLL3.7-LOX1. The lentiviral vectors (pLL3.7 and pLL3.7-LOX1) and the packaging vectors dR8.9 and VSVG were co-transfected into 293FT cells for packaging the lentivirus. H9C2 myocardial cells were infected by the lentiviruses to observe the inhibitory rate of LOX-1 on H(2)O(2)-induced apoptosis of H9C2 cells by RT-PCR, CCK-8, and Hochest33258 staining.</p><p><b>RESULTS</b>Double restriction enzyme digestion and DNA sequencing confirmed correct insertion of the sequence. Suppression of LOX-1 by the lentivirus attenuated H(2)O(2)-induced cell viability reduction and apoptosis in the myocardial cells (P<0.01).</p><p><b>CONCLUSION</b>LOX-1 activation may play an important role in H(2)O(2)-induced apoptosis of rat myocardial cells.</p>

Preparation of biotin-avidin mediated KDR-targeted liposome ultrasound contrast agent and targeted experiment in vitro / 中华超声影像学杂志

Objective To prepare a new kind of targeted liposome ultrasound contrast agent with small peptide K237 as the ligand which can combine specifically with KDR which is the main receptor of VEGF.and to test its capability in vitro. Methods Targeted bubbles(P-Bio-Av-Bio-Mbs) were formed through "biotin-avidin" bridge grafting, then they were incubated respectively with LOVO, HUVECs and LS174T which were KDR positive or negative expressed in various cells,meanwhile incubated LOVO cells with FITC- P-Bio-Av-Bio-Mbs,FITC-P-Mbs and FITC-Mbs respectively. After that, the rosette formation rate and fluorescence intensity of the combination between microbubbles and cells were observed with microscope and fluorescence microscope. After being incubated with small peptide K237 of 10 μg and 50 μg, LOVO cells were incubated with P-Bio-Av-Bio-Mbs for observing the distribution of microbubbles. Results In KDR sharply positive expressed LOVO cells, the surrounding rosette formation rate was as high as 90. 52% with the fluorescence intensity of grade 3, and it was 53. 46% with grade 2 fluorescence intensity rate in KDR positive expressed HUVECs cells, while in KDR negative expressed LS174T cells, there were few microbubbles surrounded with rosette formation rate of 5. 57% and fluorescence intensity rate of grade 0-1, therefore there were significant statistic differences in rosette formation rate among groups ( P < 0.05). After LOVO cells combined with FITC-P-Bio-Av-Bio-Mbs, FITC-P-Mbs and FITC-Mbs respectively,there were significant differences in their rosette formation rate, namely 89.62%, 7. 56% , 0 with the fluorescence rate of 3,0 - 1 and 0 respectively. Targeted cells pretreated with 10 pg K237 showed significant decreased rosette formation,and there was no formation in 50 ?g pretreated group. Conclusions KDR-Targeted liposome contrast agent with small peptide K237 liganded has been successfully prepared through biotin-avidin mediation and could combine specifically and high efficiently with targeted cells in vitro. The KDR-targeted molecular imaging of tumor neovascularizaiton may provide a new approach for early diagnosis of carcinoma.

Effect of Glycyrrhiza flavonoids on amounts of immunocytes in S180 tumor-bearing mice / 中华中医药杂志

Objective: To investigate the anti-tumor effect of Glycyrrhiza flavonoids(GF) and its immunological mechanism.Methods: Mice bearing sarcoma 180(S180) were randomized into 3 GF-treated groups and one control group.The mice in GF-treated groups were perfused with GF.Leukocyte and lymphocyte count were taken by the blood cell analyzer.Flow cytometry was performed to detect the percentages of the T cell subsets.Results: Treatment of GF resulted in the tumor inhibition rates of 52.3%(high dose group).Blood total leukocyte and lymphocyte count in GF treated groups were all higher than that in the control group,and there was the most significant increase of the number of immune cells in the high dose GF group(P

The theory and application of fluorescence resonance energy transfer / 中国病理生理杂志

The structural and functional study of protein is a major topic of current functional genomics. Fluorescence resonance energy transfer (FRET) is one of few tools available for measuring nanometer scale distances and changes in distances in vivo . FRET is an ideal technology for detection of protein conformation and protein-protein interaction by using fluorescence protein, traditional organic dyes and other dyes as probes. It uses fluorescence protein, traditional organic dyes and other dyes as its probes. The application of FRET in the determination of intracellular events would be helpful for us to understand the structure and function of biology molecules. [

Characteristics of arrhythmia symdrome in the south erea and clinical study on TCM prevention and treatment / 中医杂志

Objective: The caracteristics of rapid arrhythmia in the south region of our country are simultaneous appearance of phlegm. heat, blood stasis and deficiency, and slow arrhythmia are intermingled deficiency. blood stasis and phlegm. Thus, they were treated respectively by clearing away heat, eliminating Yang and removing blood stasis, and warming Yang to remove blood stasis respectively. Results: The total effective rate was 96. 55% in 261 cases of rapid arrhythmia treated by modified Ding Xin Decoction, and 78. 46% in the control group with a significant difference between the two groups (P

Adenoviral-mediated high efficiency expression of enhanced green fluorescence protein gene in ex vivo expanded rat mesenchymal stem cells / 中国病理生理杂志

AIM: To investigate the feasibility and infection efficiency of MSCs with replication-deficient adenovirus containing delivered gene, and whether enhanced green fluorescence protein (EGFP) gene track the change during rMSCs differentiating neuron-like cells. METHODS: Rat marrow mesenchymal stem cells (rMSCs) were expanded in low density in vitro . Under the control of CMV promoter, pAd-EGFP-Vector was constructed by homologous recombination in E.coil BJ 5183, and the recombinant virus was produced in HEK 293 packaging cell line. rMSCs infected with Ad-EGFP were observed and analyzed with fluorescence microscope. Infection efficiency was assessed by microscopical scoring and flow cytometrics. After withdrawing serum and exposure to ?-mercaptoethanol medium, rMSCs infected with Ad-EGFP was induced to differentiate into neuron-like cells. As a control, the plasmid of pTrack-EGFP also was transfected into rMSCs to evaluate transfection efficiency.RESULTS: The results showed that Adenovirus vector (AdVec) delivered EGFP gene with high efficiency to marrow mesenchymal stem cells. Gene expression analysis showed that 36%?2 % of rMSCs infected with recombinant adenovirus expressed the transgene of EGFP at high levels. However, the transfection of plasmid pTrack-EGFP using routine method of lipofectamin mixed with plasmid DNA (pTrack-EGFP) was not easily successful and the transfection efficiency was much lower. rMSCs infected with Ad-EGFP in different passage could differentiate into typical morphology alike neural cells after withdrawing serum and exposure to ?-mercaptoethanol medium. Immuno-staining with neuron-specific enolase (NSE), a neuronal marker, was strong positive, which suggested that rMSCs infected with Ad-EGFP had the potential to differentiate into neurons or neuron-like cells. CONCLUSION: The AdVec system can deliver target gene into MSCs and EGFP gene carried by AdVec can track the change during rMSCs differentiating into neuron-like cells.

Construction and expression of red fluorescent protein vectors containing different regions of human eNOS promoter / 中国病理生理杂志

AIM: To construct the plasmid vectors containing different regions of human eNOS promoter coupled to a red fluorescent protein reporter gene, which may express in mammalian cells. METHODS: Different regions of human eNOS promoter were subcloned respectively into a red fluorescent protein vector, pDsRed1-1. These recombinant vectors, pDsF1033Red, pDsF494Red and pDsF166Red, were then transfected into NIH3T3 cell lines, followed by the observation under a fluorescent microscope. RESULTS: After identified to be right by double restriction enzyme digestion, PCR and sequencing, the vectors might be effectively expressed in NIH3T3 cells. 95 % of the red fluorescent emitted by a red fluorescent protein dispersed all over the cells, appearing at 48-60 h after transfection, reaching peak at 96-144 h, becoming the strongest in light at 144 h, gradually disappearing after 168 h and remaining little red fluorescent in 21 days. The quantity and intensity in expressions of red fluorescent protein drived by different regions of human eNOS promoter were clearly lower than by a strong promoter, p CMVIE . CONCLUSION: The red fluorescent protein reporter gene vectors containing different regions of human eNOS promoter are successfully constructed and may efficaciously express in mammalian cells, appearing not strong transcriptional activities, which provide practical and feasible tools to study functions of different regions of human eNOS promoter and roles of cis-elements in it. [

Construction of recombinant adenovirus expressing BDNF and its expression in expanded rat mesenchymal stem cells in vitro / 中国病理生理杂志

AIM: To construct recombinant adenovirus vector containing brain derived neurotrophic factor, (BDNF) gene using bacterial homogenous recombination, and investigate the expression in expanded rat mesenchymal stem cells (rMSC) in vitro. METHODS: BDNF gene and proBDNF gene were subcloned into adenovirus shuttle plasmid pAdTrack-CMV containing enhanced green fluorescent protein gene (EGFP) expression cassette, forming shuttle vector of pAdTrack-BDNF, and pAdTrack-proBDNF, and co-transformed into BJ5183 bacterial cells with adenovirus backbone vector pAdEasy-1 using chemical transformation. After the recombinant adenovirus vector was obtained, the identified recombinant adenovirus plasmid DNA was digested with Pac I and transfected to 293 cells to package recombinant adenovirus particles. rMSC were infected by recombinant adenovirus and EGFP expression was detected using fluorescent microscope. Infection efficiency was assessed by flow cytometrics. Western blotting identified expression of Ad -proBDNF and Ad-BDNF in rMSC. rMSC infected with Ad -proBDNF and Ad-BDNF were induced to differentiate into neuron-like cells. rMSC infected with Ad -proBDNF and Ad-BDNF were injected into nude mice and assessd in vivo. RESULTS: We successfully constructed the recombinant adenovirus Ad -proBDNF and Ad-BDNF that expressed in expanded rMSC in vitro.CONCLUSION: Recombinant adenovirus high-effectively mediates Ad -proBDNF and Ad-BDNF expression in expanded rMSC in vitro and in vivo.