OBJECTIVETo explore the scavenging action of tenuigenin (TEN) on intracerebral amyloid β protein (Aβ) aggregation and the abnormal phosphorylated tau protein and its mechanism in Alzheimer's disease (AD) rats' brain.

METHODSAβ1-40 was injected into the right CA1 region hippocampus to establish the AD model. Successfully modeled rats were divided into the model group, the low, middle, high TEN group. Rats were administered with TEN (18.5, 37.0, 74.0 mg/kg) by gastrogavage. Besides, a sham-operation group was set up. Expression levels of Aβ1-40 and Tau p-Ser262 were detected by immunohistochemistry. Expression levels of ubiquitin (Ub) and Ub-protein ligase E3 were measured by Western blotting.The content of 26S proteasome was detected by ELISA.

RESULTSImmunohistochemical results showed that the number of Aβ and Tau p-Ser262 positively reacted neurons significantly increased in model group, when compared with the sham-operation group (P < 0.01). Results of Western blot showed expression levels of ubiquitinated protein were up-regulated and those of Ub-protein ligase E3 were down-regulated in the model group (P < 0.01). ELISA results showed that the content of 26S proteasome significantly decreased in AD rats' brain (P < 0.01). Compared with the model group, expression levels of Aβ1-40, Tau p-Ser262, and Ub significantly decreased; expression levels of Ub-protein ligase E3 apparently increased; the content of 26S proteasome significantly increased in each TEN treatment group (P < 0.05, P < 0.01). Best effect was shown in 37.0 mg/kg and 74.0 mg/kg TEN groups.

CONCLUSIONSUb proteasome pathway (UPP) participated in the occurrence of AD. TEN could obviously reduce intracere- bral Aβ1-40 accumulation and abnormal tau phosphorylation.

Alzheimer Disease ; metabolism ; Amyloid beta-Peptides ; Animals ; Drugs, Chinese Herbal ; pharmacology ; Hippocampus ; metabolism ; Neurons ; metabolism ; Phosphorylation ; Proteasome Endopeptidase Complex ; metabolism ; Rats ; Ubiquitin-Protein Ligases ; metabolism ; Ubiquitins

OBJECTIVE: To set up a natural degeneration model of chondrocytes derived from endplate of intervertebral discs of rats in order to offer an appropriate carrier for the study on mechanism of intervertebral disc degeneration. METHODS: The method of enzyme digestion combined with natural subculture was used to set up the in vitro natural degeneration model of chondrocytes derived from the endplate of intervertebral disc of rats. The morphological appearances and microstructures of the chondrocytes of different generations were observed. The expression of collagen II in chondrocytes was detected by immunocytochemical method. RESULTS: The chondrocytes derived from the endplate of intervertebral disc expressed collagen II. After 13 days of culture, the chondrocytes of generation III showed that the ability of cell division descended, the nucleoli became unclear, the cells deformed obviously, fusiform shape with weak optical activity appeared, and the intercellular space was enlarged. There were vacuoles and lipid droplets in cytoplasm. The synthesis of collagen II, as well as the cell proliferation rate, descended notably. All results showed the natural degeneration process of the chondrocytes. CONCLUSION: The in vitro natural degeneration model of chondrocytes derived from endplate of intervertebral discs of rats was successfully established. This can offer the cytological basis for study on the mechanism of intervertebral disc degeneration.

To investigate the effects of serum containing Yiqi Huayu Bushen Recipe, a compound Chinese herbal medicine, on expressions of aggrecan and type X collagen mRNAs of the degenerated intervertebral disc cells in patients with cervical spondylotic myelopathy.

To investigate the gene expression changes in the degenerated cervical intervertebral discs of rats, and to study the function of Yiqi Huayu Bushen Recipe, a compound Chinese herbal medicine, and its discomposed formulas in regulating gene expressions in the degenerated cervical intervertebral discs.

OBJECTIVE: To make an animal model of cervical spondylosis (arthralgia syndrome type) with stimulation of wind, cold, and dampness. METHODS: Twenty-four 8 months old male New Zealand white rabbits were randomly allocated into four groups: normal control group, light stimulation group, moderate stimulation group and severe stimulation group. The wind speed was 10.8-13.8 m/s, the temperature was (5+/-0.5)degrees centigrade, and the humidity was 100%. The rabbits of light, moderate, and severe stimulation groups were kept in the above-mentioned environments for 4 hours everyday, and for a total of 32, 64, and 128 hours, respectively. The intervertebral discs were stained with HE method, and observed with a light microscope. Prostaglandin E(2) (PGE(2)), 6-ketone-prostaglandin F1alpha (6-K-PGF(1alpha)) and thromboxane B(2) (TXB(2)) contents were measured by ELISA. Fas and Bcl-2 expressions were examined by immunohistochemical avidin-biotin peroxidose complex technique. Interleukin-1beta (IL-1beta), tumor necrosis factor alpha (TNF-alpha), and transforming growth factor beta (TGF-beta) mRNA expressions were examined by reverse transcription-polymerase chain reaction. RESULTS: The nucleus pulposus of rabbits in the light and moderate stimulation groups shrunken, and in the severe stimulation group, the anulus fibrosus loosed or ruptured, and the cartilage end-plate became proliferated. Compared with rabbits in the normal control group, the PGE(2) content rose in the light stimulation group, the contents of PGE(2), 6-K-PGF(1alpha), and TXB(2) increased, the expressions of IL-1beta and TNF-alpha mRNAs and Fas were up-regulated, and the expressions of TGF-beta mRNA and Bcl-2 were down-regulated in the moderate and severe stimulation groups. The expression of Fas was up-regulated mostly and Bcl-2 was down-regulated mostly in the severe group. CONCLUSION: Moderate and severe stimulations of wind, cold and dampness can lead to degeneration of cervical intervertebral discs of rabbits. The model corresponds to the theory of traditional Chinese medicine about arthralgia syndrome caused by wind, cold and dampness.

OBJECTIVE: To study the effects of Yiqi Huayu Recipe and its decomposed formulas-medicated sera on expressions of bcl-2, Bax and caspase-8 of apoptotic anulus fibrosus cells in rats. METHODS: Immunohistochemical and integral optical density analytic methods were used to observe the effects of Yiqi Huayu Recipe-, Yiqi Recipe-, Huayu Recipe-medicated sera and insulin-like growth factor 1 (IGF-1) on the expressions of bcl-2, Bax and caspase-8 of apoptotic anulus fibrosus cells in rats induced by anti-Fas antibody. RESULTS: As compared with apoptosis group, bcl-2 expression was higher, Bax and caspase-8 expressions were lower in Yiqi Huayu-treated, Yiqi-treated, Huayu-treated and IGF-1 groups (P<0.01). As compared with Yiqi-treated group and Huayu-treated group, Bax expression was lower in Yiqi Huayu-treated group (P<0.05). CONCLUSION: Yiqi Huayu Recipe and its decomposed formulas can delay degeneration of the cervical intervertebral disc, which may be due to its action in regulating the expressions of bcl-2, Bax and caspase-8.

OBJECTIVE: To study the effects of Yiqi Huayu Recipe on the gene expression pattern in normal and apoptotic chondrocytes in the cervical intervertebral disc of rats. METHODS: The intervertebral disc endplates of rats were digested enzymatically. The apoptosis of the chondrocytes was induced by anti-Fas antibody. BiostarR-40s microarray chips were used to investigate the gene expression pattern in chondrocytes in the normal control, apoptosis and Yiqi Huayu Recipe groups. The results were scanned with Scan Array 4000 and analyzed with GenePix Pro 3.0, and then subjected to standardization and ratio analysis. RESULTS: In the apoptosis group, 30 kinds of genes expressed in cervical intervertebral disc chondrocytes were screened out, in which 10 were up-regulated (ratio>2) and 20 down-regulated (ratio<0.5) as compared with the normal control group. On the other hand, in the Yiqi Huayu Recipe group, 97 kinds of genes expressed in cervical intervertebral disc chondrocytes were screened out, in which 44 were up-regulated (ratio>2) and 53 down-regulated (ratio<0.5) as compared with the apoptosis group. CONCLUSION: There are some signal transduction pathways that control the apoptosis of the intervertebral disc chondrocytes. Yiqi Huayu Recipe regulates the gene express of the apoptotic chondrocytes in intervertebral disc. The study gives a further understanding to the mechanism of the cervical spondylosis and enriches the theories of qi and blood of traditional Chinese medicine.

Objective To observe the effects of low-intensity(650 nm,5 mW)laser irradiation on the pathological changes associated with aortic atherosclerosis using a rabbit model.Methods Thirty-six male Zelanian rabbits were randomly divided into 4 equal groups:a control group,a laser irradiation group,a simvastatin treatment group and a laser plus simvastatin group,and were treated accordingly.After being fed the basic diet for a week,all the animals were fed a high fat diet during the experiment.Blood samples were taken for lipid assay at the 60th day. The animals were sacrificed at the 61st day and their aortas removed for gross and microscopic examination.Any pathological changes were graded as mild,intermediate or severe according to the indicators of atherosclerosis observed.Results There were no significant differences in blood fat levels among the four groups before the experi- ment,but significant differences in serum triglyceride prevailed after the treatments.Low-density lipoprotein(LDL)- C level in the control group was significantly higher than those in the laser irradiation,simvastatin treatment and laser plus simvastatin groups.There was also a very significant difference in high-density lipoprotein(HDL)-C levels among the laser irradiation group,the laser plus simvastatin group and the control group.The pathological changes observed were correlated with blood fat levels.Mild atherosclerosis was found in the treated groups,but severe or in- termediate atherosclerosis was more prevalent in the control group.Conclusion Low-intensity laser irradiation a- lone or combined with simvastatin can significantly decrease blood lipid levels and the severity of pathological changes associated with aortic atherosclerosis in this animal model.

Adolescent ; Adult ; Aged ; Biomarkers, Tumor ; metabolism ; Child ; Female ; Gene Expression Regulation, Neoplastic ; Hodgkin Disease ; genetics ; metabolism ; Humans ; Ki-1 Antigen ; metabolism ; Male ; Middle Aged ; PAX5 Transcription Factor ; metabolism ; Staining and Labeling ; methods ; Young Adult

Objective To evaluate the clinical application of automated urine formed elements analyzer and/or urine dipstick analyzer for examination of urinary formed elements in screening urinary tract infection (UTI). Methods 148 fresh midstream clear-catch urine samples from the UTI patients and 284 fresh midstream clear-catch urine samples from non-UTI subjects were selected. Bacteria culture was performed for bacterial colony counting and identification. Bacteria counts ( BACT), yeast-like fungus and WBC were performed by UF-looOi automated urine formed elements analyzer. Leukocyte esterase test (LEU) and nitrite test (NIT) were performed by URISYS 2400 urine dipstick analyzer. We evaluated data obtained from urine dipstick analyzer, UF-1000i and combination of UF-1000i with urine dipstick analyzer and the results was compared with those obtained from quantitative bacterial culture. Then we evaluated the sensibility, specificity, positive predictive value (PPV), negative predictive value (NPV) and accuracy. Results Among the 148 patients with UTI, the positive rate of the quantitative bacterial culture was 73.6% (109/148), the positive rate of LEU and NIT detected by dipstick test 26. 4% (39/148).There was significantly statistical difference between bacterial culture and strip test(χ2 = 55.68 ,P < 0. 05 ). The positive rate of urine flow cytometry by UF-1000i with either positive of BACT and WBC was 91.2%(135/148), which was higher than the positive rate of the quantitative bacterial culture. There was significant difference between two methods (χ2 = 14. 70, P < 0. 05 ). The positive rate of anyone positive among BACT, WBC, LEU and NIT was 94. 6% (140/148) when detected with combination of dipstick test and UF-1000i, which was higher than the positive rate of the quantitative bacterial culture. And there was significant difference between two methods (χ2 = 20. 45, P < 0. 05 ). The sensitivity of dipstick test was low (26. 4% ,39/148 ), and specificity was high ( 99. 3%, 282/284 ) . The sensitivity, specificity, positive predictive value, negative predictive value of BACT detected by UF-1000i in diagnosing urinary tract infection were 92. 6% ( 137/148 ), 39. 8% ( 113/284 ). 44. 5% ( 137/308 ) and 91.1% ( 113/124 ), respectively. If the dipstick test was combined with UF-1000i, the sensitivity, negative predictive value, specificity, positive predictive value and accuracy were 98.0% ( 145/148 ), 97.1% ( 100/103 ). 35.2% (100/284) ,44. 1% (145/329) and 56. 7% (245/432), respectively. Conclusions The combination of urine dipstick test and automated urine formed elements analyzer UF-1000i plays an important role in early diagnosis of UTI. And it has significant value in diagnosis of UTI, especially for the patients with negative bacterial cultures of urine sample.