Objective To detect the osteopontin(OPN)expression in renal tissue of rats with fluorosis and low calcium diet,and study the role of OPN in renal injury of fluorosis.Methods Forty-eight aged 1 month Wistar rats,80-120 g,were randomly divided into 4 groups by 2×2 factorial design(the number of female and male in each group was equal):the control group,high-flluoride group,low-calcium group and low-calcium with high-fluoride group.All rats of the fluorosis groups were fed with feed containing corn exposed to coal-burning from endemic fluorosis areas with high fluoride(100 mg/kg,corn),the other two groups were fed with feed containing coru from nonendemic fluorosis areas(fluoride 5 mg/kg,corn).After 16 weeks,the rats were killed.The change of teeth was examined,and the incidence rate of dental fluorosis was calculated.The expressions of both protein and mRNA of OPN in rat renal tissue were determined by RT-PCR and immunohistochemistry after four-month experimentation.Results The growth of teeth was very well in the control group and the low-calcium group.The two high-fluoride groups showed evident dental fluorosis(100%).The results of immunohistochemistry showed that the OPN protein was localized in renal tubule cytoplasm.The OPN-positive cells from renal tissue were lightly and scatteredly stained in control and low-calcium groups.The OPN-positive cells had deeper color in high-fluoride group and low-calcium with high-fluoride group,widely distributed in the renal tubular epithelial cells.The protein expression of OPN in the two groups exposed to fluoride(168.64±13.21,169.26±8.92)was significantly higher than those of the corresponding control group(145.78±10.26,all P<0.01)and low-calcium group(149.60±16.84,all P<0.01).The mRNA expression of OPN in the two groups exposed to fluoride(1.89±0.37,1.94±0.22)was significantly higher than those of the corresponding control group(1.32±0.26,all P<0.05)and low-calcium group(1.30±0.186,P<0.05),respectively.High fluoride influenced the expression of protein and mRNA of OPN(F=13.821,4.24,all P<0.05).Low calcium did not affect the expression of protein and mRNA of OPN(F=2.164,0.58,all P>0.05).However,high fluoride and low calcium had a cross interaction on the expression of protein and mRNA of OPN(F=6.257,432,all P<0.05).Conclusions Over-dose fluoride enhances the expression of OPN.The higher expression observed in the cases exposed to high fluoride concentration is associated with serious renal injury.OPN may he a potential marker for renal injury in fluorosis.Moreover,over-dose fluoride and low calcium make the renal injury worse,indicating low calcium plays an important part in renal injury by fluoride.

OBJECTIVE: To analyze 113 patients with drug-induced liver disease(DILD) so as to clinically validate the international generally-accepted diagnostic criteria.METHODS: 113 DILD cases were reviewed and categorized according to international generally-accepted diagnostic criteria and then graded separately by Maria clinical scale and DDW-J scale to analyze the correlations.RESULTS: 113 out of 822 patients with liver diseases were DILD(13.7%),among whom 17 were of hepatocellular type(15.0%),92 cholestasis type(81.5%) and 4 mixed type(3.5%).Application of Maria clinical scale and DDW-J scale led to different number of DILD cases diagnosed.DILD were mostly induced by antihyperlipidemic drugs and immunosuppressants followed by anticoagulant drugs.CONCLUSION: DDW-J scale,which is more close to clinical diagnostic criteria,contributed to the standardization of the diagnostic criteria of DILD,yet it has its limitations as well.

BackgroundPosterior capsule opacification(PCO) is the main cause inducing low vision after extacapsular cataract extraction. Our previous study determined that polylysine-ethylene diamine tetraacetic acid (EDTA) (PLE) can suppress the incidence of PCO. ObjectiveThe goal of this experiment was to investigate the inhibition of polylysine-EDTA on rabbit lens epithelial cells (LECs)proliferation in vitro and the effective concentrations of polylysine-EDTA. MethodsThe anterior capsular membranes from 10 3-month-old clean New Zealand white rabbits were digested and then cultured to obtain the LECs. The second and third generation of LECs were inoculated on the 96-hole culture plate with the cell density of the 1 × 105/ml. 12.5,25.0,50. 0,100. 0 μmol/Lof PLE were added into the culture medium for 48 hours respectively,and the DMSO medium was used at the same way as the control group. The proliferation of the LECs was then detected by MTT method and the inhibitory rate of PLE on LECs growth was calculated. ResultsLECs grew at a near normal state in ≤25.0 μmol/L PLE groups,however,cultured LECs were out of shape and the numbers decreased with the weakened adhesion ability in ≥50.0 μ mol/L PLE groups. The A490 values of LECs were 0. 278±0. 013,0. 266±0. 028,0. 260±0. 022 and 0. 247±0. 012 in 12. 5,25.0,50. 0, 100. 0 μmol/L polylysine-EDTA groups respectively and were lower than 0. 311 ±0. 038 of DMSO control group( P=0. 035,0. 011,0. 009,0.013 ). The inhibitory rates of 12. 5,25.0,50. 0, 100.0 μmoL/L PLE on LECs proliferation were 10.61％ , 14.47％ , 16.40％ and 20. 58％ respectively. ConclusionsPolylysine-EDTA can inhibit the growth and proliferation of LECs in vitro at a dose-dependent manner.

Thirteen compounds were isolated from the ethyl acetate fraction of Crepis crocea by column chromatographies on silica gel, Sephadex LH-20 and semi-preparative HPLC. The structures were elucidated on the basis of spectral analysis as tectorone I (1), 8β- (2-methyl- 2-hydroxy-3-oxobutanoyloxy) -glucozaluzanin C (2), tectoroside (3), luteolin-7-O-glucoside (4), cosmosiin (5), esculetin (6), 3,4-dihydroxybenzaldehyde (7), trans-4-hydroxycinnamic acid (8), Caffeic acid (9), methyl p-hydroxyphenyllactate (10), ethylp- hydroxyphenyllactate (11), cis-3,4-dihydroxy-β-ionion (12). All the compounds, except for compounds 4 and 9, were isolated from this plant for the first time, and tectorone I (1) is a new natural product.
Crepis ; chemistry ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Mass Spectrometry ; Molecular Structure

Objective To investigate the meaning of PURA gene and its protein in nephridial tissue of the rats with endemic fluorosis of coal burning. Methods Thirty-six SD rats of 80 - 100 g, body weight were randomly divided into control group, low fluorosis group and high fluorosis group according to body weight, 12 in each group, the number of female and male in each group was the same respectively. The control group, Low fluorosis group and high fluorosis group rots were fed with 1.5,25.0,60.0 mg/kg fluoride content in feedstuff, to establish the animal model of fluorosis. Expressions of both mRNA and its protein of PURA gene in rat nephridium tissue, were determined by RT-PCR and immunohistochemistry after four-month experimental period. Results The expressions of PURA mRNA[(2.74± 1.06),(4.29 ± 2.11)] and its protein[ (28 827.91 ± 4801.94),(61 146.96 ± 4997.55)] in low fluorosis group and high fluorosis group was higher than that in the control group[ ( 1.13 ± 0.87), (7131.95 ± 1524.54), all P < 0.05]. And the expressions of PURA mRNA and protein in high fluorosis groups was higher than that in low fluorosis greup(all P < 0.05). Conclusion High fluoride can lead to the high expression of PURA gene mRNA and protein in the rat nephridium tissue exposed to sodium fluoride.

Objective To determine clinical characteristics and the effects of drug-eluting stents on the occurrence of major adverse cardiac events during percuteneous coronary artery interventional(PCI)and long- term outcomes in patients with chronic renal insufficiency(CRI).Methods Nine hundreds and seventy three patients with angiographically-documented coronary artery disease(lumen inner diameter narrowing＞50%), included 516 patients complicated with experienced renal impairment(CRI group)and 457 with normal renal function(control group).Baseline clinical data and coronary angiographic features were recorded.Results Comparing with control group,patients in CRI group were older with higher incidence of hypertension or diabetes and simultaneously complicated by reduced left ventricular ejection fraction,and more complex coronary lesions(type C).During follow-up(mean 17 months),the mortality was significantly higher in CRI than in control group(6.2% vs 3.3%,P＜0.05),but the former with CRI was significantly lower by using drug-eluting stents in comparing with bare-metal stents(4.1% vs 8.5%,P＜0.05).Conclusion Patients with CRI often complicated with severe coronary artery disease,the mortality after PCI would be significantly reduced by using drug-eluting stents.

To study the changes of metabolites in rat urine after treatment of Aristolochia fangchi decoction by metabonomic method.

The excitability of nociceptive neurons increases in the intact dorsal root ganglion (DRG) after a chronic compression, but the underlying mechanisms are still unclear. The aim of this study was to investigate the ionic mechanisms underlying the hyperexcitability of nociceptive neurons in the compressed ganglion. Chronic compression of DRG (CCD) was produced in adult rats by inserting two rods through the intervertebral foramina to compress the L4 DRG and the ipsilateral L5 DRG. After 5-7 d, DRG somata were dissociated and placed in culture for 12-18 h. In sharp electrode recording model, the lower current threshold and the depolarized membrane potential in the acutely dissociated CCD neurons were detected, indicating that hyperexcitability is intrinsic to the soma. Since voltage-gated K(+) (Kv) channels in the primary sensory neurons are important for the regulation of excitability, we hypothesized that CCD would alter K(+) current properties in the primary sensory neurons. We examined the effects of 4-aminopyridine (4-AP), a specific antagonist of A-type potassium channel, on the excitability of the control DRG neurons. With 4-AP in the external solution, the control DRG neurons depolarized (with discharges in some cells) and their current threshold decreased as the CCD neurons demonstrated, indicating the involvement of decreased A-type potassium current in the hyperexcitability of the injured neurons. Furthermore, the alteration of A-type potassium current in nociceptive neurons in the compressed ganglion was investigated with the whole-cell patch-clamp recording model. CCD significantly decreased A-type potassium current density in nociceptive DRG neurons. These data suggest that a reduction in A-type potassium current contributes, at least in part, to the increase in neuron excitability that may lead to the development of pain and hyperalgesia associated with CCD.
Animals ; Female ; Ganglia, Spinal ; physiopathology ; Hyperalgesia ; etiology ; physiopathology ; Neurons, Afferent ; physiology ; Nociceptors ; physiology ; Pain ; physiopathology ; Potassium Channels ; physiology ; Radiculopathy ; physiopathology ; Rats ; Rats, Sprague-Dawley

The development of modern biologic techniques have provided new techniques and approaches for the modern studies on active components of Chinese medicine. This article is a review of four kinds of screening models and corresponding techniques for medicine and their applications in the studies on the active components of Chinese medicine. The four aspects comprise the whole animal models, receptor models and molecular biochromatography, cell models and cell membrane chromatography, and gene chip techniques. It will provide references for promoting studies and accelerating modernization of Chinese medicine.
Animals ; Disease Models, Animal ; Drug Evaluation, Preclinical ; methods ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; pharmacology ; Models, Molecular ; Oligonucleotide Array Sequence Analysis ; Plants, Medicinal ; chemistry

The inflammatory response is involved in the pathogenesis of the most common types of heart disease.Sanguinarine (SAN) has various pharmacological properties such as anti-inflammatory,antioxidant,antibacterial,antitumor,and immune-enhancing properties.However,few studies have investigated the effects of SAN on lipopolysaccharide (LPS)-induced inflammatory and apoptotic responses in H9c2 cardiomyocytes.Therefore,in this study,H9c2 cells were co-treated with SAN and LPS,and the mRNA levels of pro-inflammation markers and the apoptosis rate were measured to clarify the effect of SAN on cardiac inflammation.The underlying mechanism was further investigated by detecting the activation of Toll-like receptor (TLR)4/nuclear factor-κB (NF-κB) signaling pathways.As a result,increased mRNA expression of interleukin (IL)-1 β,IL-6,and TNFα induced by LPS was attenuated after SAN treatment;LPS-induced apoptosis of H9c2 cardiomyocytes and cleaved-caspase 8,9,3 were all significantly reduced by SAN.Further experiments showed that the beneficial effect of SAN on blocking the inflammation and apoptosis of H9c2 cardiomyocytes induced by LPS was associated with suppression of the TLR4/NF-κB signaling pathway.It was suggested that SAN suppressed the LPS-induced inflammation and apoptosis of H9c2 cardiomyocytes,which may be mediated by inhibition of the TLR4/NF-κB signaling pathway.Thus,SAN may be a feasible therapy to treat sepsis patients with cardiac dysfunction.