Objective To investigate the effect of Toxoplasma gondii Chinese 1 genotype infections on host brain iron metabolism and brain damages. Methods Twenty C57BL/6 mice, each weighing 15 to 17 g, were randomly divided into the control and infection groups, of 10 mice in each group. Each mouse in the infection group was injected intraperitoneally with 4 000 tachyzoites of the TgCtwh3 isolate with Chinese 1 genotype, while each mouse in the control group was injected with an equal amount of sterile phosphate-buffered saline (PBS). All mice were sacrificed 6 day post-infection and brain tissues were sampled. The iron levels were measured in mouse brain specimens using inductively coupled plasma mass spectrometry (ICP-MS). The differentially expressed genes were determined between the experimental and control groups using RNA chips and Gene Ontology (GO) term enrichment analysis of differentially expressed genes was performed. The mRNA expression of Toxoplasma gondii surface antigen 1 (TgSAG1) gene and some Zrt- and Irt-like protein (ZIP) family member coding genes was detected by quantitative real-time PCR (qPCR) assay. The ultrastructure of the hippocampus dentate gyrus in mouse brain specimens was observed using optical and electronic microscopy. The glutathione peroxidase 4 (GPx4) expression was determined using Western blotting, and malondialdehyde (MDA) level was measured using thiobarbituric acid (TBA) test. In addition, the optical density (OD) of vascular endothelial growth factor (VEGF) protein was measured using immunohistochemistry. Results Optical microscopy showed cell necrosis in the hippocampus dentate gyrus of mouse brain specimens in the infection group, and electronic microscopy cytoplasmic vacuolization, nuclear atrophy and necrosis, disruption of cristae mitochondriales and increased autophagosome levels in the mouse brain hippocampus specimens in the infection group. The iron level was significantly greater in mouse brain specimens in the infection group than in the control group [(32.92 ± 0.90) μg/g vs. (37.72 ± 1.10) μg/g; t = 3.397, P < 0.01]. RNA chips revealed 721 up-regulated genes and 276 down-regulated genes in mouse brain specimens between the infection and control groups, and the differentially expressed genes were significantly enriched in metal ion binding ability (molecular function). Elevated expression of metal element transporter ZIP2 mRNA (t = 8.659, P < 0.05), reduced GPx4 expression [(1.046 ± 0.025) vs. (0.720 ± 0.101); t = 3.129, P < 0.01], increased MDA level [(4.37 ± 0.33) nmol/mgprot vs. (5.93 ± 0.54) nmol/mgprot; t = 2.451, P < 0.05], and up-regulated mean OD of VEGF protein [(0.348 3 ± 0.017 8) vs. (0.490 6 ± 0.010 5); t = 6.641, P < 0.01] were found in mouse brain specimens in the infection group than in the control group. Conclusions Chinese 1 genotype T. gondii infection results in iron accumulation in brain tissues, reduced antioxidant ability and elevated levels of oxidative stress in mice, suggesting that T. gondii infection may cause brain damages through affecting iron metabolism in host brain tissues.

OBJECTIVETo investigate the possible effect of MicroRNA-214 (miR-214) on migration of umbilical cord blood CD34hematopoietic stem/progenitor cells induced by BM-MSC.

METHODSAfter transfection of the inhibitor or mimic of miR-214, the cultured supernatant of BM-MSC were collected respectively. The expression of miR-214 in BM-MSC was detected by real time quantitative PCR, the effect of supernatant on CD34cell migration was evaluated by chemotaxis assays. The levels of chemokine(SDF-1) secreted by BM-MSC in the supernatant were detected by ELISA.

RESULTSThe cultured supernatant of BM-MSC could promote the migration of CD34cells. Compared with the group without transfection or negative control(NC) group, the transfection with miR-214 mimic could promote the migration of CD34cells (P<0.01), while the migration rate in miR-214 inhibitor groups decreased significantly (P<0.01). Further study found that the concentration of SDF-1 was not changed notably in all groups (P>0.05), as compared with the control group.

CONCLUSIONmiR-214 signalings may indirectly increase the migration of CD34hematopoietic stem/progenitor cells by modulating BM-MSC functions, which may not significantly correlate with the chemokine SDF-1 secreted by BM-MSC.

This study was aimed to investigate the influence of TLR2 and TLR4 agonists on the migration and adhesion activity of human bone marrow-derived mesenchymal stem cells (MSC) and to clarify the underlying mechanisms. The expression of TLR2 and TLR4 on MSC was detected by flow cytometry. The effects of TLR2 agonist (PAM3CSK4) and TLR2 agonist (LPS) on MSC migration and adhesion ability were evaluated with chemotaxis and adhesion test. The results indicated that expressive levels of TLR2 and TLR4 on surface of human bone marrow MSC were (24.5 ± 3.2)% and (91.3 ± 5.2)% respectively. Compared with the control group, the migration activity of MSC toward SDF-1 was decreased significantly in PAM3CSK4 group, while the adhesion activity of MSC was promoted by PAM3CSK4 exposure. However, both the migration activity toward SDF-1 and the adhesion activity of MSC were not changed significantly in LPS-treated group. Further, it was found that PAM3CSK4 did not affect the expressive level of CXCR4 on MSC, however, it could inhibit the spontaneous migration of MSC in dose dependent manner. It is concluded that activation of TLR2 can decrease the migration ability of MSC, which may associate with the decreased spontaneous migration ability and the increased adhesion activity of MSC.
Bone Marrow Cells ; cytology ; drug effects ; Cell Movement ; drug effects ; Cells, Cultured ; Humans ; Lipopeptides ; pharmacology ; Lipopolysaccharides ; pharmacology ; Mesenchymal Stromal Cells ; cytology ; drug effects ; Toll-Like Receptor 2 ; agonists ; Toll-Like Receptor 4 ; agonists

BACKGROUNDThe renoprotective mechanisms of adenosine monophosphate (AMP)-activated protein kinase (AMPK) agonist - metformin have not been stated clearly. We hypothesized that metformin may ameliorate inflammation via AMPK interaction with critical inflammatory cytokines. The aim of this study was to observe the effects of metformin on expression of nuclear factor-κB (NF-κB), monocyte chemoattractant protein-1 (MCP-1), intercellular adhesion molecule-1 (ICAM-1) and transforming growth factor-beta 1 (TGF-β1) induced by high glucose (HG) in cultured rat glomerular mesangial cells (MCs).

METHODSMCs were cultured in the medium with normal concentration glucose (group NG, 5.6 mmol/L), high concentration glucose (group HG, 25 mmol/L) and different concentrations of metformin (group M1, M2, M3). After 48-hour exposure, the supernatants and MCs were collected. The expression of NF-κB, MCP-1, ICAM-1, and TGF-β1 mRNA was analyzed by real time polymerase chain reaction. Western blotting was used to detect the expression of AMPK, phospho-Thr-172 AMPK (p-AMPK), NF-κB p65, MCP-1, ICAM-1, and TGF-β1 protein.

RESULTSAfter stimulated by HG, the expression of NF-κB, MCP-1, ICAM-1, TGF-β1 mRNA and protein of MCs in group HG increased significantly compared with group NG (P < 0.05). Both genes and protein expression of NF-κB, MCP-1, ICAM-1, TGF-β1 of MCs induced by high glucose were markedly reduced after metformin treatment in a dose-dependent manner (P < 0.05). The expression of p-AMPK increased with the rising of metformin concentration, presenting the opposite trend, while the level of total-AMPK protein was unchanged with exposure to HG or metformin. Conlusion Metformin can suppress the expression of NF-κB, MCP-1, ICAM-1 and TGF-β1 of glomerular MCs induced by high glucose via AMPK activation, which may partly contribute to its reno-protection.

AMP-Activated Protein Kinases ; metabolism ; Animals ; Cells, Cultured ; Glomerular Mesangium ; cytology ; Glucose ; pharmacology ; Mesangial Cells ; drug effects ; metabolism ; Metformin ; pharmacology ; NF-kappa B ; metabolism ; Rats

Objective To investigate the necessity of preoperative use of prophylactic antibiotics in percutaneous transluminal angioplasty (PTA) of lower extremity. Methods A total of 86 patients with arteriosclerosis obliterans of lower extremity (101 invalid lower extremities in total) were enrolled in this study. The patients were prospectively and randomly divided into study group (n=41, 51 limbs) and control group (n=45, 50 limbs). The patients in the study group received intravenously prophylactic antibiotics two hours before PTA, while no antibiotic was employed for the patients in the control group. The improvement of symptoms and the occurrence of infection after PTA in the two groups were compared. Results After PTA, fever was seen in 27 patients, including 12 patients of the study group (29.3%) and 15 patients of the control group (33.3%). Elevation of neutrophil count (>70%) was observed in 6 patients (14.7%) of the study group and in 7 patients (15.6%) of the control group, but the difference between the two groups was not statistically significant (P>0.05). Septicemia occurred in one patient in each group, both were aged patients with diabetes. The post-treatment infection rate in the study group and in the control group was 1.96% and 2.00%respectively, the difference between the two groups was not significant (P>0.05). Conclusion There is no significant correlation between the use of prophylactic antibiotics and the infections after PTA of lower extremity. Therefore, the clinical value of using prophylactic antibiotics for patients with high risk of infection needs to be verified by further randomized controlled trials.

Exosomes are naturally nanometer-sized (40-100 nm) vesicles that contribute to the communication among the cells through the proteins, lipids and RNA which they carried. Exosomes derived at different physiological conditions play different role, which therefore can be widely used in the diagnosis and treatment of cancer. Besides, exosomes as the natural nanomaterials have the unique advantage in drug delivery system. Although the intrinsic advantages of exosomes have evoked a surge of interest, improvements in standardized isolation techniques with high efficiency and robust yield are still a huge challenge. Here, we provide an overview of the isolation and the application of exosomes in drug delivery system.

Objective Using the influenza virus hemagglutinin (HA) as the target to screen for novel anti-influenza polypeptide drugs. Methods The HA binding peptides were screened out through affinity selection from a 12-peptide phage library, and the anti-H1N1 activity was evaluated at MDCK cell and chicken embryo(ovo). Results Nine HA binding peptides were finally obtained, and the H6 peptide was found having significant antiviral activity against H1N1. Its IC50 against two strains of H1N1, A/FM1/1/47 (H1N1) and A/PPR8/34(H 1 N 1), were 37. 3 and 48. 5 μmoZL respectively determined by cytopathic effect (CPE)test, and 26. 7 and 33. 4 μmoI/L respectively measured by ovo antiviral experiment. Conclusion These results showed that H6 might be a potential herapeu icdrug for H1N1 infecion.

Objective: To establish an extraction process optimization method based on fingerprint combined with principal component analysis, which was finally applied to Compound Huzhang ethanol extraction process optimization. Methods: Taking Compound Huzhang prescription as model drug, different ethanol extraction condition fingerprint was established by HPLC, harvesting the areas of the common peaks, the total factor scores were calculated by PCA. Arranged experiments with U9(93×31) uniform design method, choosing the total factor scores as index, the influence of extration times (X1), alcohol consumption(X2), alcohol concentration(X3), and time of extracting(X4) on the yield of extract was investigated, then the technological parameters of optimum ethanol extraction condition were selected by multi-nonlinear mathematic models. Results: Multi-nonlinear mathematic models described the relationship between response indexes and factor variables with a regression coefficient of 0.997. The optimized conditions for Compound Huzhang Prescription were extracted in 90% ethanol six times as much as it for three times, 1h once. Conclusion: It is proved that the extraction method is suitable and feasible that could provide a reference for compound Chinese medicine extraction process optimization.

ObjectiveTo investigate the protective effect of Wuziyanzong Pills (WYP) in the rat model of oligoasthenospermia (OAS) and its action mechanism.

METHODSSixty male SD rats were equally randomized into six groups: normal control, OAS model, Shengjing Capsules (1.6 g per kg of the body weight), low-dose WYP (1 g per kg of the body weight), medium-dose WYP (2 g per kg of the body weight), and high-dose WYP (4 g per kg of the body weight). The OAS model was established by intragastric administration of Tripterygium glucoside at 30 mg per g per d for 6 weeks. From the 3rd week of modeling, the rats of the medication groups were treated intragastrically with corresponding drugs for 4 weeks. Then all the rats were sacrificed for measurement of the testicular and epididymal organ coefficients, examination of epididymal sperm quality and apoptosis, and detection of the openness of the sperm mitochondrial permeability transition pore (MPTP). Histopathological changes in the testis were observed by HE staining and the apoptosis of spermatogenic cells determined by Hochest staining.

RESULTSWYP obviously improved the organ coefficients of the testis and epididymis, increased sperm concentration, motility and viability, decreased the apoptosis of spermatogenic cells, and inhibited the abnormal openness of MPTP in the OAS model rats. HE staining showed that the number and levels of spermatogenic cells were significantly increased while Hochest staining manifested that the apoptosis of spermatogenic cells was remarkably inhibited in the seminiferous tubules of the testis in the WYP-treated rats.

CONCLUSIONSWYP can improve sperm quality and reduce the apoptosis of spermatogenic cells (including sperm) in OAS model rats, which may be related with its inhibitory effect on the abnormal openness of MPTP.

Animals ; Apoptosis ; drug effects ; Asthenozoospermia ; chemically induced ; drug therapy ; Drugs, Chinese Herbal ; pharmacology ; Epididymis ; drug effects ; Male ; Mitochondrial Membrane Transport Proteins ; drug effects ; Oligospermia ; drug therapy ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Sperm Count ; Sperm Motility ; drug effects ; Spermatozoa ; cytology ; drug effects ; Testis ; drug effects ; Tripterygium

Based on the experience of our team combined with literature analysis, there were some problems in study of clinical intervention, mainly in the aspects of clinical trial registration, construction of intervention programs, recruitment and compliance of research objects, intervention fidelity, selection of outcome indicators and data collection and analysis. It is necessary to understand the importance of clinical trial registration and follow the registration procedure, establish an intervention program based on the theoretical framework, focus on the factors that impede or promote the recruitment of objects to ensure the sample size, formulate strategies to improve the compliance and intervention fidelity of the intervention objects and select the appropriate outcome indicators， apply appropriate data collection and analysis methods based on the content of the study, and thus improve the quality of intervention researches.