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OBJECTIVE@#To investigate the antidepressant-like effects of Chaihu Shugan Powder (CSP, ) and to explore its underlying mechanisms.@*METHODS@#Thirty-two Sprague-Dawley rats were randomly divided into control (CON), chronic unpredictable mild stress (CUMS), fluoxetine (FLU), and CSP groups, 8 rats in each group. All of the rats except for those in the control group were subjected to 3 consecutive weeks of CUMS to establish the depression model. The open field test (OFT), forced swimming test (FST), and sucrose preference test were used to assess the anti-anxiety and antidepressant effects of CSP. Terminal deoxynucleotidyl transferase (TdT) dUTP nick-end labeling was used to determine the apoptosis rate in the hippocampal tissues. The mRNA and protein levels of glucose-regulated protein (GRP) 78, spliced X-box-binding protein (XBP)-1, CCAAT/enhancer-binding protein homologous protein (CHOP), caspase-12, and c-Jun N-terminal kinase (JNK) in the hippocampus of rats were evaluated by real-time PCR and Western blot analysis, respectively.@*RESULTS@#Administration of CSP alleviated anxiety and depression-like behavior in CUMS rats, as revealed by enhanced time and distance in the center of the OFT (P<0.05), an increased preference for sucrose, and longer swimming time and shorter immobility time during the FST (all P<0.05). In addition, CSP treatment significantly reduced the rate of apoptosis in rat hippocampal neurons (P<0.05). The mRNA and protein expression levels of GRP78, spliced XBP-1, and CHOP were down-regulated along with the expression of caspase-12 and cleaved caspase-12 proteins (all P<0.05), whereas total and phosphorylated JNK1 protein levels did not differ significantly between control and CSP-treated rats.@*CONCLUSION@#CSP can improve depression-like behavior in rats exposed to CUMS, possibly by suppressing CHOP and caspase-12 mediated apoptosis in the rat hippocampus.
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Buffaloes , Cells, Cultured , In Vitro Techniques , Primary Cell Culture , Stem Cells , TretinoinABSTRACT
Currently, the systems for culturing buffalo spermatogonial stem cells (SSCs) in vitro are varied, and their effects are still inconclusive. In this study, we compared the effects of culture systems with undefined (foetal bovine serum) and defined (KnockOut Serum Replacement) materials on the in vitro culture of buffalo SSC-like cells. Significantly more DDX4- and UCHL1-positive cells (cultured for 2 days at passage 2) were observed in the defined materials culture system than in the undefined materials system (p < 0.01), and these cells were maintained for a longer period than those in the culture system with undefined materials (10 days vs. 6 days). Furthermore, NANOS2 (p < 0.05), DDX4 (p < 0.01) and UCHL1 (p < 0.05) were expressed at significantly higher levels in the culture system with defined materials than in that with undefined materials. Induction with retinoic acid was used to verify that the cultured cells maintained SSC characteristics, revealing an SCP3⺠subset in the cells cultured in the defined materials system. The expression levels of Stra8 (p < 0.05) and Rec8 (p < 0.01) were significantly increased, and the expression levels of ZBTB16 (p < 0.01) and DDX4 (p < 0.05) were significantly decreased. These findings provided a clearer research platform for exploring the mechanism of buffalo SSCs in vitro.
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Currently, the systems for culturing buffalo spermatogonial stem cells (SSCs) in vitro are varied, and their effects are still inconclusive. In this study, we compared the effects of culture systems with undefined (foetal bovine serum) and defined (KnockOut Serum Replacement) materials on the in vitro culture of buffalo SSC-like cells. Significantly more DDX4- and UCHL1-positive cells (cultured for 2 days at passage 2) were observed in the defined materials culture system than in the undefined materials system (p < 0.01), and these cells were maintained for a longer period than those in the culture system with undefined materials (10 days vs. 6 days). Furthermore, NANOS2 (p < 0.05), DDX4 (p < 0.01) and UCHL1 (p < 0.05) were expressed at significantly higher levels in the culture system with defined materials than in that with undefined materials. Induction with retinoic acid was used to verify that the cultured cells maintained SSC characteristics, revealing an SCP3⺠subset in the cells cultured in the defined materials system. The expression levels of Stra8 (p < 0.05) and Rec8 (p < 0.01) were significantly increased, and the expression levels of ZBTB16 (p < 0.01) and DDX4 (p < 0.05) were significantly decreased. These findings provided a clearer research platform for exploring the mechanism of buffalo SSCs in vitro.
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Near-infrared spectroscopy ( NIR ) is widely used in the area of food quantitative and qualitative analysis.Variable selection technique is a critical step of the spectrum modeling with the development of chemometrics.In this study, a novel variable selection strategy, automatic weighting variable combination population analysis (AWVCPA), was proposed.Firstly, binary matrix sampling (BMS) strategy that gives each variable the same chance to be selected and generates different variable combinations, was used to produce a population of subsets to construct a population of sub-models.Then, the variable frequency ( Fre) and partial least squares regression ( Reg) , which were two kinds of information vector ( IVs) were weighted to obtain the value of the contribution of each spectral variables, the influence of two IVs of Rre and Reg was considered to each spectral variable.Finally, it used the exponentially decreasing function ( EDF) to remove the low contribution wavelengths so as to select the characteristic variable.In the case of near infrared spectrum of beer and corn, the prediction model based on partial least squares ( PLS ) was established.Compared with other variable selection methods, the research showed that AWVCPA was the best variable selection strategy in the same situation.It had 72.7% improvement compared AWVCPA-PLS with PLS and the predicted root mean square error (RMSEP) decreased from 0.5348 to 0.1457 on beer dataset.It had 64.7% improvement compared AWVCPA-PLS with PLS and the RMSEP decreased from 0.0702 to 0.0248 on corn dataset.
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Hyoscyamine and scopolamine are two main alkaloids in Atropa belladonna with great medicinal value. In this paper, the contents of hyoscyamine and scopolamine, the upstream products in alkaloid synthesis, and the expression levels of key enzyme genes PMT, TRⅠ and H6H in secondary metabolism of A. belladonna seedlings were measured to clarify the mechanism of nitrogen forms regulating alkaloids synthesis.The results showed that the 50/50 (NH⁺₄/NO⁻₃) treatment was more favorable for the accumulation of alkaloids and the conversion of hyoscyamine to scopolamine. The content of putrescine was almost consistent with the change of key enzymes activities in the synthesis of putrescine, they both increased with the rise of ammonium ratio, reaching the highest at 75/25 (NH⁺₄/NO⁻₃). The detection of signaling molecule nitric oxide (NO) showed that the NO concentration decreased with the decrease of nitrate proportion. Further detection of gene expression levels of PMT, TRⅠ and H6H in TAs synthesis pathway showed that a certain amount of ammonium promoted the expression of PMT and H6H in roots. When the ratio of ammonium to nitrate was 50/50, PMT, TRⅠ and H6H in leaves and roots had higher expression levels. It can be speculated that the regulation of the formation of hyoscyamine to scopolamine by nitrogen forms mainly through affecting the expression of key enzyme genes. 50/50 (NH⁺₄/NO⁻₃) treatment increased the gene expression of TRⅠ in both leaves and roots as well as PMT and H6H in roots, promoting the synthesis of putrescine to hyoscyamine and the conversion of hyoscyamine to scopolamine.
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Atropa belladonna , Genetics , Gene Expression Regulation, Plant , Hyoscyamine , Mixed Function Oxygenases , Nitrogen , Metabolism , Scopolamine , MetabolismABSTRACT
Objective To investigate the incidence and risk factors of surgical site infection(SSI) in patients with internal fixation surgery for limb fracture.Methods Medical data of patients with internal fixation surgery for limb fracture in a hospital from January 2013 to January 2016 were collected, 39 patients with SSI following internal fixation was as infection group, according to the 1:2 ratio, 78 patients without SSI following operation during the same period were randomly selected as the control group, risk factors of SSI were analyzed.Results Among 4 125 patients undergoing internal fixation surgery, incidence of SSI was 0.95% (n=39), the positive rate of bacterial culture in infection group was 87.2% (34/39), a total of 38 strains of pathogenic bacteria were isolated, among which 22 were gram-positive strains (57.9%), 15(39.5%)were gram-negative strains,1(2.6%) was fungi,Staphylococcus aureus was the main pathogenic bacteria (47.4%), and there were 20 isolates of multidrug-resistant organisms.Univariate analysis showed that infection group and control group was significantly different in the following aspects: combined underlying diseases, time from injury to operation≥8 hours, open fracture, multiple fracture, duration of operation≥180 minutes, intra-operative blood loss≥400 mL, allogeneic blood transfusion, duration of postoperative indwelling drainage tube≥5 days, and average length of hospital stay≥14 days (all P<0.05).Multivariate logistic regression analysis showed that the following factors were risk factors for SSI following internal fixation surgery for fracture: time from injury to operation≥8 hours, open fracture, duration of operation≥180 minutes, duration of postoperative indwelling drainage tube≥5 days, and average length of hospital stay≥14 days (all P<0.05).Conclusion Risk factors for SSI in patients with internal fixation surgery for limb fracture are multiple, reducing risk factors has a positive effect on decreasing the incidence of SSI and improving the cure rate.
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<p><b>OBJECTIVE</b>To study the effect of Tiantai No. 1 [symbol in text] on gene expression profile in hippocampus of Alzheimer's disease (AD) rat, molecular genetic target points of the effect of this drug were defined, its molecular genetic pharmacodynamic mechanism of anti-AD was further explored at molecular gene level, and a scientific basis was provided for its clinical availability and promotion.</p><p><b>METHODS</b>Thirty male Sprague-Dawley rats were divided into three groups with 10 rats per group: sham-operation group, model group and Tiantai No. 1 group. Sterile surgical procedure was applied, the model group with bilateral hippocampal injection of Aβ1-40 was established, and normal saline was used instead of Aβ1-40 in the sham-operation group. One week after the models was made, rats were administered by gastric lavage once every day for three consecutive weeks. The rats of the sham-operation group and the model group were daily fed with purified water by lavage; the rats of the Tiantai No.1 group treated group were administered with Tiantai No.1 by lavage. Total RNAs of hippocampus tissues were extracted with Trizol, the changes of hippocampus gene expression profiles in the above three groups were analyzed by using Affymetrix rat whole genome expression profile microarray.</p><p><b>RESULTS</b>Microarray analysis showed that, compared with the sham-operation group, the hippocampus of the model group had 50 up-regulated genes with significant difference (fold change >2), and 21 down-regulated genes with significant difference (fold change <0.5); compared with the hippocampus of the model group, the hippocampus of the Tiantai No. 1 group was found to have 5 up-regulated genes with significant difference (fold change >2) and 20 down-regulated genes with significant difference (fold change <0.5). The functions of differentially expressed genes of the groups were involved in nervous system's development, neuronic differentiation and function-regulation, cellular growth and differentiation and apoptosis, synaptic occurrence and plasticity, inflammation and immune response, ion channels/transporters, cellular signal transduction, cellular material/energy metabolism and so on.</p><p><b>CONCLUSION</b>Tiantai No. 1 can regulate hippocampal function, and further regulate the brain function of animals in multiple gene target points by a number of ways.</p>
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Animals , Male , Alzheimer Disease , Genetics , Pathology , Body Weight , Computational Biology , Methods , Drugs, Chinese Herbal , Pharmacology , Electrophoresis, Agar Gel , Gene Expression Profiling , Gene Expression Regulation , Hippocampus , Metabolism , Pathology , Nucleic Acid Denaturation , Organ Size , RNA , Metabolism , Rats, Sprague-DawleyABSTRACT
<p><b>OBJECTIVE</b>To investigate the chemical constituents in leaves of Ilex centrochinensis and their antitumor bioactivity.</p><p><b>METHOD</b>Various chromatography techniques such as column chromatography on silica gel, Sephadex LH-20 and preparative HPLC were used to isolate and purify the compounds and their structures were identified by spectral data and physicochemical properties. Their antitumor effect was tested by MTT method.</p><p><b>RESULT</b>Ten compounds were isolated and identified as 1,4-benzenediol (1), (2S)-5,4'-dihydroxy-7,3'-dimethoxyflavan(2), (2S)-5,4'-dihydroxy-7-methoxyflavan (3), kaempferol (4), quercetin (5), naringenin (6), ursolic acid (7), uvaol (8), oleanolic acid (9) and beta-sitosterols (10).</p><p><b>CONCLUSION</b>Compounds 1-5, 7, 8 were isolated from the species for the first time, among which compounds 1-3 were isolated from the Ilex genus for the first time. Compounds 2 and 3 showed strong cytotoxic activity against Huh7 cell lines with IC50 values of 8.98, 13.04 mg x L(-1), respectively. Compounds 7-9 exhibited weak cytotoxic activity against Caco-2 cell lines with IC50 values of 28.52, 38.28, 33.04 mg x L(-1), respectively.</p>
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Humans , Antineoplastic Agents , Chemistry , Pharmacology , Caco-2 Cells , Cell Line, Tumor , Cell Survival , Dose-Response Relationship, Drug , Ilex , Chemistry , Inhibitory Concentration 50 , Plant Extracts , Chemistry , Pharmacology , Plant Leaves , Chemistry , Plants, Medicinal , ChemistryABSTRACT
The SRY gene from buffalo (Bubalus bubalis) genome was amplified by the polymerase chain reaction ( PCR) with primers based on the sequence of Hostein SRY gene. The amplified fragment was 2005 bp include 5UTR ( 1 - 504bp) and 3'UTR(1196 - 2005bp). And the amplified fragment was cloned and sequenced. Sequence analysis showed that the coding region of SRY gene (505 - 1195bp) from buffalo was highly homologous with those of other bovine counterpart genes (96% homology) , especially in the HMG box region (99%homology). It was found that there were only signal on male buffalo genome on Southern blot,which indicate SRY gene are highly conservative on evolves.