Zhao L.,Guiyang Medical University |
Yu Y.,Guiyang Medical University |
Deng C.,Guiyang Medical University
Toxicology Letters | Year: 2014
In order to investigate the Sonic hedgehog (Shh) signaling pathway and the effect of cyclopamine in rat hepatocytes with chronic fluorosis, 48 Wistar rats were randomly divided into 4 groups. The control group was provided with tap water in which the fluorine concentration was <1. mg/L, while the remaining three groups were provided with water containing sodium fluoride (NaF) at a concentration of 50. mg/L. After 6 months, the blocking and blocking control groups were injected intraperitoneally once every 2 days for 6 days with 10. mg/kg cyclopamine or dimethyl sulfoxide, respectively. The urinary and skeletal fluoride contents were determined by the ion selective electrode method. Levels of aspartate transaminase (AST), alanine transaminase (ALT), total protein (TP) and albumin (Alb) in the serum were determined by using autobiochemical machine. Histological changes in liver tissue were evaluated with Hematoxylin & Eeosin (H&E) staining using light microscopy. The protein and mRNA expression of Shh, Smo and Gli1 in hepatocytes of experimental animals was determined by immunohistochemistry (IHC), Western blotting (Wb) and Real-time quantitative PCR (RT-qPCR). Fluoride content of the urine and bone was increased in the fluorosis and blocking groups compared to those in the control group (P<. 0.05), while fluoride content in the blocking group was decreased compared to the fluorosis and blocking control groups (P<. 0.05). The expression of Shh, Smo and Gli1 at the mRNA and protein levels was significantly increased in hepatocytes from the fluorosis and blocking control groups compared with the control group, and expression in the blocking group was lower than that of the fluorosis and blocking control groups. The difference between any two groups was considered to be statistically significant (P<. 0.05). Taken together, our study indicates that the expression of Shh, Smo and Gli1 at the protein and mRNA level in hepatocytes of rats with chronic fluorosis can be increased by fluoride and may be inhibited by cyclopamine and that the Shh signaling pathway plays an important role in the liver pathogenesis caused by fluorosis. © 2014 Elsevier Ireland Ltd.
Xiao C.-D.,Guiyang Medical University |
Shen X.-C.,Guiyang Medical University |
Tao L.,Guiyang Medical University
International Journal of Pharmaceutics | Year: 2013
Solvent evaporation/extraction method is widely used for core-shell microspheres fabrication. However, the solvent evaporation rate, as an essential factor for polymers phase separation, is difficult to control which results in failure of complete phase separation between polymers. At the present study, the selective dissolution technique was used to improve the phase separation, and successfully fabricate core-shell microspheres for controlled delivery of drug with reduced initial burst release. The core-shell microspheres were prepared with poly(L-lactic-co-glycolic acid) (PLGA) and poly(L-lactic acid) (PLLA), and aspirin was used as model drug. Ethyl acetate (EtAc) was applied to ameliorate the phase separation during the preparation process. The ratio of dichloromethane (DCM)/EtAc seriously affected the distribution of polymer molecules and the formation of the core-shell structure. The internal morphology of the microspheres varied depending on the amount of EtAc. Core-shell structure with a dense core of PLLA and a shell of PLGA was well formed when 2 ml EtAc was used. The differential scanning calorimeters (DSC) results showed two distinct melting points which confirmed a completely polymer phased separation occurred. These microspheres showed sustained release of aspirin for at least 456 h with a little burst release (3.49%). © 2013 Elsevier B.V. All rights reserved.
Liu Y.-J.,Guiyang Medical University |
Guan Z.-Z.,Guiyang Medical University |
Gao Q.,Guiyang Medical University |
Pei J.-J.,Karolinska Institutet
Toxicology Letters | Year: 2011
In order to reveal the mechanism of the brain injury induced by chronic fluorosis, the levels of apoptosis and c-Jun N-terminal kinases (JNK) in brains of rats and SH-SY5Y cells exposed to different concentrations of sodium fluoride (NaF) were detected. The dental fluorosis and fluoride contents in blood, urine and bones of rats were measured to evaluate the exhibition of fluorosis. The apoptotic death rate was measured by flow cytometry and the expression of JNK at protein level by Western blotting. The results showed that as compared with controls, the apoptotic death rate was obviously increased in brains of the rats exposed to high-fluoride (50. ppm) for 6 months with a concentration dependent manner, but no significant change for 3 months. In SH-SY5Y cells treated with high concentration (50. ppm) of fluoride, the increased apoptotic death rate was obviously observed as compared to controls. In addition, the expressions of phospho-JNK at protein level were raised by 20.5% and 107.6%, respectively, in brains of the rats exposed to low-fluoride (5. ppm) and high-fluoride for 6 months; while no significant changes were found between the rats exposed to fluoride and the controls for 3 months. The protein level of phospho-JNK was also increased in SH-SY5Y cells exposed to high-fluoride. There were no changes of total-JNK both in the rats and in the SH-SY5Y cells exposed to excessive fluoride as compared to controls. When SH-SY5Y cells were singly treated with SP600125, an inhibitor of phospho-JNK, the decreased expression of phospho-JNK, but no apoptosis, was detected. Interestingly, after JNK phosphorylation in the cultured cells was inhibited by SP600125, the treatment with high-fluoride did not induce the increase of apoptosis. In addition, there was a positive correlation between the expression of phospho-JNK and the apoptotic death rate in rat brains or SH-SY5Y cells treated with high-fluoride. The results indicated that exposure to excessive fluoride resulted in the increase of apoptosis in rat brains and SH-SY5Y cells, in which one of the mechanisms might be activating JNK phosphorylation. © 2011 Elsevier Ireland Ltd.
Zeng Y.,Guiyang Medical University |
Song J.-X.,Chinese University of Hong Kong |
Shen X.-C.,Guiyang Medical University
Phytotherapy Research | Year: 2012
Atherosclerosis (AS) is a systemic cardiovascular disease with complicated pathogenesis involving oxidative stress, endothelial dysfunction and chronic inflammation. Increasing lines of evidence have questioned the statins-dominated treatment for AS, including their dangerous side-effects such as the breakdown of muscle when taken in larger doses. A multifaceted approach that addresses all major risk factors or pathological targets may provide an ideal treatment for AS. Studies of the herbal remedies on the prevention and treatment of AS have received much attention in recent years. This review summarizes some important experimental findings regarding their mechanisms of action on AS. Using the pre-set PUBMED searching syntax and inclusion criteria, representative citations published in English concerning the experimental studies of 14 herbal materials were included. We found that many extracts and (or) single compounds from these herbal materials, such as Salvia miltiorrhiza, Curcuma longa, Rheum undulatum and Panax notoginseng, could regulate multiple key targets involved in the initiation and propagation of AS. Some important findings about the effects of herbal formulations on AS were also reviewed. Given the complicated nature of AS and the holistic, combinational approach of herbal remedies, we propose that mixed herbal preparations with multiple active ingredients may be preferable for the prevention and treatment of AS. Further rigorously designed pharmacological evaluation and multi-centred clinical trials are warranted. © 2011 John Wiley & Sons, Ltd.
Zhou W.,Guiyang Medical University |
Zhang X.-Y.,Hubei University of Medicine
American Journal of Chinese Medicine | Year: 2013
Radix isatidis (R. isatidis) (Banlangen) is a traditional Chinese medicine (TCM) famous for its broad antiviral activity. Its clinical medical history spans several thousands of years in China. Many scientists and scholars have conducted systematic research on this herb from its pharmacognosy to pharmaceuticals, especially in China. Through our research and literature reports, we inferred that the antiviral activity of R. isatidis mostly depended on the water-soluble part, including amino acids, IRPS, nucleosides, and sulfur-containing alkaloids. By playing a role in directly killing pathogenic viruses or regulating the immune system to enhance anti-virus ability, R. isatidis's biological activities mostly depend on the synergistic effect of its multiple components. This article aims to expand understanding of R. isatidis in the following aspects including medicinal resources, chemical constituents, pharmacological activities, clinical applications, and separation and analytical technologies. © 2013 World Scientific Publishing Company Institute for Advanced Research in Asian Science and Medicine.
Yao M.,Guiyang Medical University
Zhonghua yu fang yi xue za zhi [Chinese journal of preventive medicine] | Year: 2013
OBJECTIVE: To establish coal arsenic poisoning rat model by feeding the rats with the corn powder baked by high arsenic coal as the main raw material.METHODS: Fifty Wistar rats, healthy, were randomly divided into 5 groups according to the figures of their weights, including control group, drinking arsenic poisoning water group, low, medium and high arsenic contaminated grain group, 10 rats for each.Rats in control group and drinking arsenic poisoning water group were fed with standard feed without any arsenic containing. Rats in water group would drink 100 mg/L As2O3 solution and the rats in arsenic grain groups would be fed with the arsenic contaminated grain at the dose of 25, 50 and 100 mg/kg, respectively. The duration would last for 3 months.General situation and weight were observed. At the same time, the arsenic contents of urine, hair, liver and kidney of the rats in each group were detected, as well as the histopathology changes of liver and kidney, and the ultra structure of liver was observed.RESULTS: The arsenic contents of urine (median(min-max)) of the rats in the arsenic water group, low, medium and high arsenic grain groups were separately 3055.59 (722.43-6389.05), 635.96(367.85-1551.31), 1453.84 (593.27-5302.94) and 3101.11 (666.64-6858.61) µg/g Cr; while the arsenic contents of hair of the rats in the above groups were separately (23.07 ± 10.38), (8.87 ± 3.31), (12.43 ± 6.65) and (25.68 ± 7.16) µg/g; the arsenic contents of liver of the rats in the above groups were separately (5.68 ± 3.13), (2.64 ± 1.52), (3.89 ± 1.76) and (5.34 ± 2.78) µg/g; and the arsenic contents of kidney were separately (6.90 ± 1.94), (3.48 ± 1.96), (5.03 ± 2.08) and (7.02 ± 1.62) µg/g; which were all significantly higher than those in the control group (86.70 (49.71-106.104) µg/g Cr,(1.28 ± 0.37) µg/g, (1.01 ± 0.34) µg/g and (1.82 ± 1.09) µg/g, respectively). The difference showed significance (P < 0.05). Under electron microscope detection, we observed the reduction of mitochondrial, the blurred mitochondrial cristae, some disappeared ridges, the reduced rough endoplasmic reticulum, and irregular uneven nuclear in the liver cells of rats in arsenic contaminated grain group. The contents of aspartate transaminase (AST) and total bile acid (TBA) in medium and high arsenic contaminated grain group were respectively (196.17 ± 46.18), (212.40 ± 35.14) U/L and (11.74 ± 4.07), (19.19 ± 4.68)µmol/L, which were higher than it in the control group (separately (143.10 ± 29.13) U/L and (6.23 ± 2.95)µmol/L). The contents of glutathione-S-transferases(GST), γ-glutamyltranspeptidase (GGT)and blood urea nitrogen (BUN)in high arsenic contaminated grain group were separately (196.21 ± 47.38)U/L, (1.71 ± 0.66)U/L, (9.54 ± 1.95)mmol/L, which were higher than that in the control group ((134.93 ± 24.80 )U/L, (0.75 ± 0.36)U/L, (7.67 ± 1.02)mmol/L, respectively). The contents of cholinesterase (CHE) in low, medium and high arsenic contaminated grain group were separately (259.90 ± 52.71)U/L, (263.44 ± 66.06)U/L and (244.90 ± 36.14)U/L, the contents of total protein(TP) in rats of high arsenic contaminated grain group were (62.64 ± 5.50)g/L, which was all lower than that in the control group ((448.33 ± 59.67)U/L, (69.38 ± 4.24)g/L, respectively). The contents of TBA in high arsenic contaminated grain group ( (19.19 ± 4.68) µmol/L) was higher than that in drinking water arsenic poisoning group ((15.15 ± 2.64)µmol/L). The differences of the above indexes were all significant (P < 0.05).CONCLUSION: The results showed the arsenic poisoning rat model produced by coal-burning were successfully established.
Hong F.,Guiyang Medical University
Zhonghua yu fang yi xue za zhi [Chinese journal of preventive medicine] | Year: 2013
OBJECTIVE: To observe the chronic combined effects of sodium fluoride and sodium arsenite on the Runx2 and downstream related factors of bone metabolism in SD rats.METHODS: SD rats were divided randomly into nine groups of 6 each by factorial experimental design (half female and half male) , and supplied with the different doses of fluoride, arsenite and fluoride plus arsenite containing in deionized water (untreated control containing 0 mg/kg fluoride and 0 mg/kg arsenite, and low-fluoride and high supplemented with 5 and 20 mg/kg fluoride, and low-arsenite and high supplemented with 2.5 and 10 mg/kg arsenite, and low-fluoride plus low-arsenite, and low-fluoride plus high-arsenite, and high-fluoride plus low-arsenite, and high-fluoride plus high-arsenite, respectively) . After 6 months exposure, the concentration of Runx2, matrix metallopeptidase 9 (MMP-9) ,Osterix, Receptor activator for nuclear factor-κ β ligand (RANKL) were detected by enzyme-linked immunosorbent assay method, respectively.RESULTS: There were no dental fluorosis found in the control group, low-arsenic group and high-arsenic group. There were significant differences in the constituent ratio of dental fluorosis among the rats from low-fluoride and high-fluoride (that is 5 rats out of 6 and 6 rats out of 6) compared with the control group (0 rat out of 6) (χ(2) = 8.57, 12.00, P < 0.05). The bone fluorine level increased with the increase of fluoride dose, the groups without fluoride supply (control group, low-arsenite and high-arsenite group's geometric mean (minimum-maximum) were 0.005 (0.003-0.009), 0.006 (0.003-0.021), 0.003 (0.002-0.100) mg/g, respectively), low-fluorine groups (low-fluoride group, low-fluoride plus low-arsenite, and low-fluoride plus high-arsenite group were 3.395 (2.416-5.871), 3.809 (1.471-7.799), 1.471 (1.473-6.732)mg/g, respectively) , the high-fluorine groups (high-fluoride, high-fluoride plus low-arsenite, and high-fluoride plus high-arsenite group were 70.086 (46.183-131.927), 69.925 (40.503-96.183), 40.503 (52.622-89.487) mg/g, respectively) and the differences between groups was significant (P < 0.05). The bone arsenic level increased with the increase of arsenite dose. The low-arsenic groups (low-arsenite group, low-arsenite plus low-fluoride, and low-arsenite plus high-fluoride group were 7.195 (5.060-9.860), 6.518 (2.960-12.130), 6.970 (3.400-9.730) µg/g, respectively), the high-arsenic groups (high-arsenite, high-arsenite plus low-fluoride, and high-fluoride plus high-arsenite group's geometric mean(minimum-maximum) were 8.823 (5.760-10.840), 9.470 (7.230-12.860), 8.321 (2.420-17.540) µg/g, respectively) were significantly higher than that in the groups without arsenic supply (control group, low-fluoride and high-fluoride group were 1.785 (0.300-3.750), 2.226 (1.410-3.980), 2.030 (1.040-3.850)µg/g, respectively) (P < 0.05). There was no significant difference of the bone arsenic concentration between low-arsenic and high arsenic group. There was significant positive correlation between fluoride concentration and Runx2, MMP-9, Osterix, RANKL level (the correlation coefficient was 0.647, 0.354, 0.582, 0.613 between fluorine gavage concentration and protein level, the correlation coefficient was 0.559,0.387, 0.487, 0.525 between bone fluorine concentration and protein level, respectively, P < 0.01). There was negative correlation between arsenite gavage concentration with Runx2 level (r = -0.527, P < 0.05) and was no correlation between arsenite gavage concentration with MMP-9, RANKL,Osterix level (P > 0.05). There was interaction between fluoride and arsenite to Runx2, MMP-9, RANKL,Osterix (F = 3.88, 15.66, 2.92, 6.42, respectively, P = 0.01, <0.01, 0.031, <0.01, respectively).CONCLUSION: The combined effects of fluoride and arsenic on the Runx2, MMP-9, RANKL, Osterix of bone metabolism showed antagonistic effects.
Luo W.,Guiyang Medical University
Zhongguo xiu fu chong jian wai ke za zhi = Zhongguo xiufu chongjian waike zazhi = Chinese journal of reparative and reconstructive surgery | Year: 2012
To construct a new type of self-assembling peptide nanofiber scaffolds-RGDmx, and to study the cell compatibility of the new scaffolds and the proliferation and chondrogenic differentiation of precartilaginous stem cells (PSCs) in scaffolds. PSCs were separated and purified from newborn Sprague Dawley rats by magnetic activated cell sorting and indentified by immunohistochemistry and immunofluorescent staining. The RGDmx were constructed by mixing KLD-12 and KLD-12-PRG at volume ratio of 1:1. PSCs at passage 3 were seeded into the KLD-12 scaffold (control group) and RGDmx scaffold (experimental group). The proliferation of PSCs in 2 groups were observed with the method of cell counting kit (CCK)-8 after 1, 3, 7, and 14 days after culture. The RGDmx were constructed by mixing KLD-12-PRG and KLD-12 at different volume ratios of 0, 20%, 40%, 60%, 80%, and 100% and the proliferation of PSCs was also observed. The complete chondrogenic medium (CCM) was used to induce chondrogenic differentiation of PSCs in different scaffolds. The differentiation of PSCs was observed by toluidine blue staining and RT-PCR assay. PSCs were separated and purified successfully, which were identified by immunohistochemistry and immunofluorescent staining methods. The results of CCK-8 showed that the absorbance (A) value in the experimental group increased gradually and reached the highest at 7 days; the A value in the experimental group was significantly higher than that in the control group at 7 days and 14 days (P < 0.05). Meanwhile, the A value in the RGDmx scaffold with a volume ratio of 40% was significantly higher than those in others (P < 0.05). After 14 days of induction culture with CCM, the toluidine blue staining results were positive in 2 groups; the results of RT-PCR showed that the expression levels of collagen type II and the aggrecan in the experimental group were significantly higher than those in the control group (P < 0.05). The self-assembling peptide nanofiber scaffold-RGDmx is an ideal scaffold for tissue engineer because it has good cell compatibility and more effective properties of promoting the differentiation of PSCs to chondrocytes.
Chen J.,Guiyang Medical University
Ceramics International | Year: 2015
As one of the most potential negative electrode materials, Na2Ti6O13 is expected to play an important role in the area of high-performance battery. In this work, we have developed an easy, efficient and controllable method to prepare rod-shaped Na2Ti6O13 crystals. This approach utilized a single-source molten salt strategy and only needed to sinter a special precursor synthesized from an aqueous solution containing H3BO3 and (NH4)2TiF6 in presence of sodium salts. The component and shape of precursor crystals can be tuned by adjusting the reagent concentration and reaction temperature. By sintering precursor crystals in air at 900 °C for 30 min, Na2Ti6O13 with high crystallinity and purity can be obtained. X-ray diffraction and scanning electron micrographs results of different sintering times show that the sintering process can be divided into two steps. Firstly, the precursor crystals are converted to TiO2 (anatase) nano-particles and amorphous sodium salts. Subsequently, molten salt reaction occurs between amorphous sodium salts and TiO2 and forms rod-shaped Na2Ti6O13 crystals. © 2015 Elsevier Ltd and Techna Group S.r.l. All rights reserved.
Yang T.-T.,Guiyang Medical University
Chinese Journal of Endemiology | Year: 2013
Objective: To investigate DNA methylation in the promoter region, mRNA transcription and protein expression of glutathione-S-transferases-P1 (GSTP1) gene and their relation with arsenism. Methods: In endemic coal-pollution-borne arsenism area, Jiaole village of Xinren county, Guizhou province, according to the diagnostic criteria of endemic arsenism(WS/T 211-2001), 123 cases with endemic arsenism were selected and divided into three groups (mild arsenism group: 42 cases, moderate arsenism group: 41 cases and severe arsenism group: 40 cases). Forty seven residents were selected as controls in a village about 12 km away from the endemic arsenism area. With the informed consent principle, peripheral blood of all respondents was collected in order to analyze DNA methylation and check mRNA. DNA methylation of GSTP1 gene promoter region in peripheral blood was assayed by PCR, and GSTP1 mRNA expression was assayed using real-time quantitative PCR. In addition, other cutaneous specimens originated from 53 cases with arsenism that accepted surgical treatment voluntarily were taken. Of these specimens, general pathological changes were 28 cases, precancerous 20 cases and cancerous 5 cases. Skin tissues of 15 cases of non-tumor surgery patients without abnormal pathological changes were as control group. GSTP1 protein expression in the skin tissue was detected using immunohistochemistry (IHC). Results: Among different groups of arsenic poisoning, the positive rate of DNA methylation of GSTP1 gene was 28.57%(12/42) in the mild group, 57.10%(23/41) in the moderate group and 65.00%(26/40) in the severe group. Compared with the control group (6.38%,3/47), the difference was statistically significant (χ2 = 7.792, 26.000, 33.412, all P < 0.01). Among different groups of arsenic poisoning diagnosed by dermapathology, the positive rate of DNA methylation of GSTP1 gene was 21.43%(6/28) in the general pathological change group, 50.00%(10/20) in the precancerous group and 80.00%(4/5) in the cancerous group. Compared with the control group(6.67%,1/15), the difference was statistically significant (χ2 = 3.562, 7.468, 10.756, all P < 0.05). It showed that the positive rate of DNA methylation of GSTP1 gene increased with aggravation of the disease and dermatic lesion of arsenism(tendency χ2 = 38.239, χ2.= 13.659, all P < 0.01). Compared with the control group(0.184 26), the expressions of GSTP1 mRNA in peripheral blood in moderate (0.087 77) and severe arsenic poisoning groups(0.056 93) were significantly reduced(all P < 0.01), and that of severe group was significantly lower than that of the moderate group (P < 0.01); compared with the control group (0.338 45) and the general lesion group (0.276 74), GSTP1 mRNA expression was significantly reduced in precancerous lesion group(0.104 81) and cancerous group(0.043 70), in which the cancerous group was significantly lower than that of the precancerous lesions. The difference of skin tissue GSTP1 protein expression rate between groups was statistically significant (χ2 = 20.948, P < 0.05), in which the difference between the precancerous lesion group(65.00%, 13/20), the cancer group (40.00%, 2/5) and the control group (100.00%, 15/15) was statistically significant (χ2 = 12.183, 11.778, P < 0.01). Spearman correlation analysis showed that the degree of skin lesion and the level of GSTP1 protein expression was negatively correlated(r = - 0.520, P < 0.05). Groups were divided according to DNA methylation of GSTP1 gene, and the mRNA and protein expression of GSTP1 in methylation group(0.038 40, 57.14%) was significantly lower compared with that of unmethylated group(0.187 07, 95.74%; Z = 9.032, χ2 = 23.134, all P < 0.01). Conclusions: Arsenism may lead to DNA methylation of human GSTP1 gene promoter region, thereby inhibiting expression of mRNA and protein. GSTP1 gene plays an important role in arsenism or carcinogenic process.