Effects of Ginsenoside Rg1 on the Biological Activity of Human Periodontal Ligament Cells.

, Objective. To investigate the effect of ginsenoside Rg1 on the biological activity of primary cultured human periodontal ligament cells (PDLC). Methods. The effects of ginsenoside Rg1 on the proliferation activity, protein synthesis, and alkaline phosphatase (ALP) activity of primary cultured human periodontal ligament cells were investigated by thiazole blue (MTT) colorimetric method, Coomassie brilliant blue method, and enzyme kinetics method. The effect of ginsenoside Rg1 on cell cycle was detected by flow cytometry, and the cells were labeled with calcium ion-sensitive fluorescent probe Fluo3/AM, and the effect of ginsenoside Rg1 on intracellular free calcium concentration was detected by laser scanning confocal microscope. Results. Compared with the control group, the experimental groups of ginsenoside Rg1 at various concentrations could significantly promote cell proliferation, and the effect time was the longest in the concentration range of 0.01-0.05 µmol/L;, Rg1 0.01umol/L and 0.05umol/L. The protein content in the 72-hour cell culture medium of the µmol/L group was significantly higher than that of the control group; the ALP activity in the 72-hour cell culture medium of the Rg1 0.01 µmol/L, 0.05 µmol/L, and 0.1 µmol/L groups was significantly higher than that of the control group; FCM assay showed that after 0.1 µmol/L Rg1 for 48 hours, compared with the control group, the proportion of cells in the early stage of DNA synthesis (G1%) of PDLC was significantly reduced, while the proportion of cells in the DNA synthesis stage (S%) and the value of cell proliferation index PrI (S + G2M)% were significantly increased; Rg1 increased intracellular calcium in PDLC cells at first and then decreased and finally maintained at a slightly higher resting calcium level than before drug addition. Conclusion. Ginsenoside Rg1 can increase the proliferation activity, protein synthesis, and alkaline phosphatase activity of periodontal ligament cells within a certain concentration range; Rg1 reduces the cells in G1 phase and increases cells in S phase of periodontal ligament fibroblasts. Change the concentration of free calcium ions in cells and promote more cells to enter a proliferative state.

Stabilization of microtubules improves cognitive functions and axonal transport of mitochondria in Alzheimer's disease model mice.

One major pathological process in Alzheimer's disease is mediated by hyperphosphorylated tau, which includes altered microtubules (MTs) and functions associated with tau. A potential way to compensate for altered MT function is to use an MT stabilizer, such as epothilone D (EpoD). Previous studies have demonstrated improved cognitive functions and axonal transport by EpoD in tau-mutation mice. Here, we demonstrated that extended EpoD treatment also has beneficial effects on APP/PS1 double-transgenic mice, improving their motor and spatial memory, increasing key synaptic protein levels, while not affecting amyloid plaque density or level of tau phosphorylation. Interestingly, EpoD appears to improve the retrieval of formed memories. We also observed improved axonal transport of mitochondria in cultured neurons from APP/PS1 mice. In addition, higher level of perineuronal nets are found in APP/PS1 mice injected with EpoD, suggesting potential contributions of increased inhibition. Our results suggest potential therapeutic value of EpoD in treating Alzheimer's disease.

Chinese Herbal Medicine Xueshuantong Enhances Cerebral Blood Flow and Improves Neural Functions in Alzheimer's Disease Mice.

Reduced cerebral blood flow in Alzheimer's disease (AD) may occur in early AD, which contributes to the pathogenesis and/or pathological progression of AD. Reversing this deficit may have therapeutic potential. Certain traditional Chinese herbal medicines (e.g., Saponin and its major component Xueshuantong [XST]) increase blood flow in humans, but whether they could be effective in treating AD patients has not been tested. We found that systemic XST injection elevated cerebral blood flow in APP/PS1 transgenic mice using two-photon time-lapse imaging in the same microvessels before and after injection. Subchronic XST treatment led to improved spatial learning and memory and motor performance in the APP/PS1 mice, suggesting improved neural plasticity and functions. Two-photon time lapse imaging of the same plaques revealed a reduction in plaque size after XST treatment. In addition, western blots experiments showed that XST treatment led to reduced processing of amyloid-ß protein precursor (AßPP) and enhanced clearance of amyloid-ß (Aß) without altering the total level of AßPP. We also found increased synapse density in the immediate vicinity of amyloid plaques, suggesting enhanced synaptic function. We conclude that targeting cerebral blood flow can be an effective strategy in treating AD.

Interaction network analysis of differentially expressed genes and screening of cancer marker in the urine of patients with invasive bladder cancer.

OBJECTIVE: To detect the expression profile of bladder cancer and to delineate the interaction network of these genes in invasive bladder cancer. METHODS: A total of 126 differentially expressed genes were identified, and input into STRING online database to delineate interaction network. The network data were screened with central nodes. The expression of genes with the most evident change in the exfoliated cells of urine was detected. RNA markers with over-expression in stage Ta tumor and/or T1 to T4 tumors but low expression in blood or inflammatory cells were characterized. RESULTS: On the basis of assay of 21,639 whole-genome oligonucleotide microarray, a total of 126 differentially expressed genes were identified, of which 69 had up-regulated expression and 57 had down-regulated expression. STRING screening showed there were interactions among 103 genes in the bladder cancer which formed a complex network. A total of 23 central nodes were screened with Cytoscape and are involved in multiple signaling pathways related to tumorigenesis. The test specificity was 80% for the 30 control patients with urinary tract infections. The combination of BLCA-4 and HOXA13 could distinguish between low and high grade tumors, with specificity and sensitivity of 80%. CONCLUSION: The interaction network of differentially expressed genes, especially the central nodes of this network, can provide evidence for the early diagnosis and molecular targeted therapy of invasive bladder cancer, and combined detection of IGF-1, hTERT, BLCA-4 and HOXA13 genes is helpful to evaluate BTCC at different stages.

Screening and identification of specific markers for bladder transitional cell carcinoma from urine urothelial cells with suppressive subtractive hybridization and cDNA microarray.

OBJECTIVE: The objective of this study was to screen and identify differentially expressed genes in invasive bladder transitional cell carcinoma (BTCC). METHODS: Voided urine samples were collected from consecutive patients with BTCC and patients under surveillance for bladder cancer recurrence; voided urine samples from patients with non-malignant diseases served as control. We identified the differentially expressed genes by comparing urine samples of bladder carcinoma to that of the control group with suppressive subtractive hybridization (SSH) and cDNA microarray. The differentially expressed genes were verified by quantitative real-time polymerase chain reaction (QPCR). RESULTS: From the 762 white colonies, a total of 449 positive clones were obtained in which 112 were found to be upregulated in BTCC. Sequencing and homology analysis were performed for these 112 clonies. The detection rates of some known genes (including IGF-1, human telomerase reverse transcriptase [hTERT], bladder cancer specific nuclear matrix protein 4 [BLCA-4] and homeobox A13 [HOXA13]) for BTCC at the Ta, T1 and >T1 stages were 48%, 90% and 100%, respectively, with a specificity of 85%. The test specificity was 80% for the 30 control patients with urinary tract infections. The combination of BLCA-4 and HOXA13 could distinguish between low- and high-grade tumours, with specificity and sensitivity of 80%. CONCLUSION: We successfully constructed a reliable SSH library of BTCC and found that combination detection insulin-like growth factor 1 (IGF-1), hTERT, BLCA-4 and HOXA13 genes could help to evaluate BTCC at different stages.

[Tumor necrosis factor alpha-308 polymorphism and asthenospermia].

OBJECTIVE: To investigate the relationship between -308 genotype polymorphism in the promoter region of the tumor necrosis factor alpha (TNFalpha) gene and asthenospermia in infertile men. METHODS: Allele-specific polymerase chain reaction (ASPCR) and polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) were used to analyze the genotype at position -308 in the promoter region of the TNFalpha gene in 187 infertile male patients, who were divided into Groups A (asthenospermia, n = 60), B (oligoasthenozoospermia, n = 65) and C (infertile patients with normal sperm, n = 62). The levels of TNFalpha in the seminal plasma from these patients were measured by radioimmunoassay, and all the data were statistically analyzed by SPSS16.0. RESULTS: Groups A and B exhibited significant differences from C in the frequency of GA/AA at position 308 in the promoter region of the TNFalpha gene (21.67% and 26.15% versus 8.06%, P < 0.05). Spearman analysis showed a negative correlation between the GA + AA type of the TNFalpha-308 allele and the percentage of grade a + b sperm (r = -0.690, P < 0.05). The level of TNFalpha in the seminal plasma was significantly elevated in Groups A ([4.23 +/- 0.45] ng/ml) and B ([4.29 +/- 0.47] ng/ml) as compared with C ([4.03 +/- 0.66] ng/ml, P < 0.05), but with no significant differences between Groups A and B (P > 0.05). It was also significantly higher in the GA+AA ([4.61 +/- 0.29] ng/ml) than in the GGtype ([4.06 +/- 0.45] ng/ml, P < 0.05). CONCLUSION: Regardless of sperm density, the frequently of TNFalpha-308 GA/AA is negatively correlated with the percentage of grade a + b sperm, which may be associated with the level of TNFalpha in the seminal plasma. Accordingly, anti-TNFalpha therapy might be effective for asthenospermia, and the measurement of the TNFalpha level in the seminal plasma can be an auxiliary diagnostic marker for male infertility.