Objective To observe the effects of wild jujube seed decoction (WJSD) on learning memory and the levels of malondialdehyde (MDA) and the activities of supemxide dismutase (SOD) and nitric oxide synthase (NOS) in the brain of the pathogeny model rats with sleep deprivation (SD). Methods Levels of learning memory and MDA, and activities of SOD and NOS in rats' brain were assayed after SD induced by made-self multiple platform method ( MMPM), senescence induced by D-galactose, and Yin and blood-deficiency induced by eyclophosphamide and hydrocortisonum. Results After 48 h SD, the levels of learning memory was lower in the model group( ( 108.9 ± 12.5 ) s, ( 89 ± 11.5 ) s, ( 0 ) ) than those in the environmental control group ( ( 47.3 ±4.6)s,(9±1.4)s,(6.5 ±1.2))(( t=4.36,3.18,2.07, P＜0.01＝＝. While MDA, and activities of SOD and NO in rats'brain higher in the model group( (3.8 ±0.6) ,(3.0 ±0.5)nmol · mgprot-1 ,(229.7 ±25.8) ,(236.3± 25.2 ) U · mgprot - 1, ( 5.7 ± 0. 8 ), ( 5.4 ± 0.9 ) U · mgprot - 1 ) than those in the environmental control group ( (2. 1 ±0.4) ,(1.6 ±0.4)nmol · mgprot-1 ,(155.5 ±10.6) ,( 147.2 ±26.1 )U · mgprot-1 ,(2.8 ±0.7),(2.9 ± 0.5 ) U · mgprot -1 ) ( t = 2.89,3.01,6.78,5.94,3.10,3.46, P ＜ 0.05 ＝. However, the observation of the groups treated with WJSD, the levels of learning memory showed a tendency in returning to normal level (P ＜ 0.05 ＝ ,and MDA, and activities of SOD and NO of the high dose of WJSD showed a tendency in returning to normal leve1 (P ＜ 0.05 ＝ ,and the low dose of WJSD showed a tendency in returning to normal level too(P ＜ 0.05 ＝. Conclusion WJSD can improve the disability of learning and memory of the pathogeny rats model, and its one of mechanism maybe involve the reduction of neural cell damage with free radical and NO.

OBJECTIVE: To study the role of nuclear factor-kappa B (NF-kappaB) and cycloxygenase-2 (COX-2) in the onset of phlegm obstruction due to lung-deficiency in rats and the therapeutic mechanism of Huatan Recipe. METHODS: Twenty-four SD rats were randomly divided into normal group, model group and treatment group, with 8 rats in each group. The rats in the model group and treatment group were exposed to sulfur dioxide and cold wind to establish the rat model of phlegm obstruction due to lung-deficiency, and the rats in the treatment group were also treated with Huatan Recipe, a compound traditional Chinese medicine. The expression of NF-kappaB in the bronchial epithelial cells of the rats was tested with the method of immunohistochemistry, and the COX-2 mRNA in the lung tissues of the rats was measured by using reverse transcription-polymerase chain reaction. RESULTS: The expressions of NF-kappaB and COX-2 mRNA in rats of the model group were higher than those of the normal group (P<0.01), and the expressions of NF-kappaB and COX-2 mRNA in rats of the treatment group were obviously lower than those of the model group (P<0.01). CONCLUSION: The NF-kappaB and COX-2 play an important role in the onset of phlegm obstruction in rats. Huatan Recipe may prevent the development of phlegm obstruction by down-regulating the expressions of NF-kappaB and COX-2 mRNA.

Objective To observe the effects of wild jujube seed decoction(WJSD) on ultrastructure and astrocytes expression in the brain cortex of old model rats with blood-deficiency and Yin and sleep deprivation(SD)after SD. Methods Ultrastructural changes in cortical parts of the experiment rat were observed by transmission electron microscopy , immunohistochemical staining was used to detect star-shaped glial cells marker-glial fibrillary acidic protein(GFAP) expression after SD induced by made-self multiple platform method (MMPM), senescence induced by D-galactose, and Yin and blood-deficiency induced by cyclophosphamide and hydrocortisonum. Results Compared to environmental control group((9.8 ± 2.5), (0.11 ± 0.02) ) , syndrome model group (( 20.4 ±4.4), (0.20 ±0.011) ) rats had obvious ultrastructural changes and stronger expression of GFAP in cortical parts (t = 7.63,3.18, P ＜ 0. 01), while those of W J SD of high and low dose group ((14.4 ± 3.9), (15.5 ± 6.4),(0.14 ±0.02)(0.14±0.02)) rats showed weaker(t=6.32,5.24,2.31,2.45, P＜0.05). Conclusion WJSD of large and small lose group could improve the rats neurons pathological changes,WJSD downward adjusting the expression of GFAP may be one of the mechanisms of treatment on insomnia with blood-deficiency and Yin in the old.

OBJECTIVE: To observe the effects of Guben Huatan Tongmai Recipe (GBHTTMR), a compound Chinese herbal recipe, on expressions of macrophages and cell adhesion molecules (CAMs) of aortic endothelia in rats with syndrome of phlegm blocking blood vessel, and to explore the pathogenesis of the phlegm-pathogen. METHODS: Fifty normal male Wistar rats, 7-week in age, were randomly divided into five groups: normal control group, untreated group, high-dose GBHTTMR-treated group, low-dose GBHTTMR-treated group and simvastatin-treated group, with 10 rats in each group. Syndrome of phlegm blocking blood vessel was induced in rats of the latter 4 groups by feeding the rats with high lipid diet. Levels of blood lipid were compared among the 5 groups. The expressions of macrophages and CAMs in aortic endothelia were tested by immunohistochemical staining method. RESULTS: The level of blood lipid, and the expressions of macrophages and CAMs showed statistical differences between the normal control group and the untreated group (P<0.01), and between the untreated group and the low-, high-dose GBHTTMR-treated and simvastatin-treated groups as well (P<0.05). CONCLUSIONS: GBHTTMR can decrease the level of serum cholesterol and triglycerides, and increase the level of high density lipoprotein. It also can inhibit the expressions of macrophages, intercellular adhesion molecule-1, vascular cell adhesion molecule-1, P-selectin, and E-selectin.

Objective To design the small interference RNA (siRNA) specific to human CCL20 gene by RNA interfering technique, construct its recombinant lentiviral expression vectors, and identify these vectors by DNA sequencing. Methods According to Tuschl’s principle, the siRNA was designed and converted into cDNA of shRNA (small hairpin RNA) of siRNA for CCL20 gene. The cDNA was synthesized and inserted into plasmid pHSER-dsRNA-GFP-SIN which was linearized by restriction endonucleases SpeⅠ and SalⅠ. The recombinant plasmid was transformed into competent E. coli. DH5? cells. The positive recombinant colony was selected by ampicillin medium agar and identified by DNA sequencing. Results Two recombinant lentiviral plasmids of siRNA specific to CCL20 gene were constructed successfully. Their DNA sequence analysis completely coincided with their designed sequences. Conclusion Lentiviral vector-based siRNA expression plasmids against CCL20 gene have been successfully constructed and identified. They will be further used for interfering CCL20 mRNA transcription and lay the foundation for CCL20 gene modified human keratinocyte stem cells.

Objective:To investigate the complex Calculus Bovis-target-keratitis network and to explore the molecular mechanism of Calculus Bovis treating keratitis through network pharmacology. Methods:Genes related to keratitis were searched in the online DisGeNET database and the protein-protein interaction (PPI) network of keratitis-associated proteins was constructed.The components isolated and identified in Calculus Bovis were collected through the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP, https: //tcmsp-e.com/tcmsp.php), Chemistry Database by Shanghai Institute of Organic Chemistry of CAS (http: //www.organchem.csdb.cn), and published literature.The canonical SMILES information of the collected components was exported, which were submitted to the SwissTargetPrediction platform to predict potential targets of the components.The active component-predicted target network of Calculus Bovis was constructed and merged with the PPI network of keratitis-associated proteins to build the active component-potential target network of Calculus Bovis and systemically investigate the potential targets and signal pathways of Calculus Bovis in treatment of keratitis.The component-target-pathway network was established to analyze the mechanism of Calculus Bovis treating keratitis. Results:Thirty-nine components isolated and identified in Calculus Bovis were searched and 65 target genes related to keratitis were screened.Of the 28 potential targets involved in Calculus Bovis treating keratitis, there were 7 direct targets, including tumor necrosis factor, caspase 1, Toll-like receptor 9, C-X-C motif chemokine ligand 8, interleukin-6, mitogen-activated protein kinase 8, neurotrophic receptor tyrosine kinase 1.The 28 potential targets were annotated to 12 entries for biological process, 18 for cellular components and 13 for molecular function.In the Kyoto encyclopedia of genes and genomes pathway enrichment analysis, 10 signal pathways were identified as enriched categories, which were mainly related to human cytomegalovirus infection, amoebiasis, antifolate resistance, PI3K-Akt signaling pathway, rheumatoid arthritis, apoptosis, cytokine-cytokine receptor interaction, malaria, non-alcoholic fatty liver disease, interleukin-17 signaling pathway. Conclusions:Calculus Bovis may play an adjuvant therapeutic effect on keratitis through anti-inflammatory, antibacterial, antiviral, immune regulation, inflammatory regulation and other functions.

ObjectiveTo investigate the effect and mechanism of Shenling Baizhusan on the treatment of oligoasthenospermia with hyperuricemia （HUA）. MethodThirty-two male Kunming （KM） mice were randomly divided into blank group （n=6）， model group （n=6）， high-dose Shenling Baizhusan group （n=7）， low-dose Shenling Baizhusan group （n=7）， and febuxostat group （n=6）. Except for the blank group， all other groups received intraperitoneal injection of potassium oxazinate suspension （600 mg·kg^-1） for 7 days. After modeling， the high-dose Shenling Baizhusan group and the low-dose Shenling Baizhusan group were orally administered with 20.14 g·kg^-1 and 10.07 g·kg^-1 of Shenling Baizhusan， respectively. The Febuxostat group was orally administered with 0.25 g·kg^-1 of Febuxostat， while the blank group and model group were orally administered with the same volume of physiological saline. Oral administration was performed once a day for 14 consecutive days， after which samples were collected. Biochemical methods were used to measure serum uric acid （UA）， superoxide dismutase （SOD） and malondialdehyde （MDA） in testicular tissue. Hematoxylin-eosin （HE） staining was used to observe the histopathological changes in testicular tissue and evaluate the spermatogenesis function. Automated sperm analyzer was used to measure sperm density and motility. Single-cell gel electrophoresis （SCGE） was used to assess sperm DNA integrity. Terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling （TUNEL） was used to detect testicular cell apoptosis rate. Western blot analysis was performed to measure the protein expression levels of Kelch-like ECH-associated protein 1 （Keap1）， nuclear factor E₂-related factor 2 （Nrf2）， heme oxygenase-1 （HO-1）， B-cell lymphoma-2 （Bcl-2）， Bcl-2-associated X protein （Bax）， and Caspase-3 in testicular tissue. Real-time polymerase chain reaction （PCR） was conducted to evaluate the mRNA expression levels of Keap1， Nrf2， and HO-1 in testicular tissue. ResultCompared with the blank group， the model group showed elevated serum UA level （P<0.01）， decreased testicular spermatogenesis function， sperm density， and motility （P<0.01）， and increased sperm trailing rate and testicular cell apoptosis rate （P<0.01）. Compared with the model group， the high-dose Shenling Baizhusan group showed significant improvements in the above-mentioned indicators （P<0.05， P<0.01）. Additionally， the expression levels of Keap1， Bax， and Caspase-3 in testicular tissue were reduced， while the expression levels of Nrf2， HO-1， and Bcl-2 increased （P<0.05， P<0.01）. The mRNA level of Keap1 decreased （P<0.05， P<0.01）， while the mRNA levels of Nrf2 and HO-1 increased （P<0.05， P<0.01）. ConclusionShenling Baizhusan can significantly improve HUA oligoasthenospermia， and its mechanism may be related to the Nrf2/antioxidant response element （ARE） signaling pathway.