Objective To investigate the effect of serum containing Huangqi decoction on the proliferation, migration, and tubulogenesis of rat liver sinusoidal endothelial cells (LSECs) induced by vascular endothelial growth factor (VEGF) and its mechanism of action based on the protein kinase B (AKT)/mammalian target of rapamycin (mTOR) signaling pathway. Methods The serum containing Huangqi decoction was prepared, and rat LSECs were isolated and cultured in vitro . Then rat LSECs were randomly divided into blank group, VEGF group, serum control group, and low-, middle-, and high-dose serum containing Huangqi decoction groups. MTT colorimetry, Transwell assay, and tubulogenesis assay were used to measure the proliferation, migration, and tubulogenesis abilities of LSECs in each group, and Western Blot was used to measure the protein expression levels of platelet endothelial cell adhesion molecule-1 (CD31), endothelin-1 (ET-1), and endothelial nitric oxide synthase (eNOS), as well as AKT, phosphorylated-AKT (p-AKT), mTOR, and phosphorylated mTOR (p-mTOR) in the AKT/mTOR signaling pathway. A one-way analysis of variance was used for comparison of continuous data between multiple groups, and the Tukey's test was used for further comparison between two groups. Results Compared with the blank group, the VEGF group and the serum control group had significantly promoted proliferation, migration, and angiogenesis of rat LSECs (all P < 0.05), and Western Blot showed significant increases in the expression levels of CD31, ET-1, eNOS, and AKT/mTOR signaling pathway-related proteins (all P < 0.05). There were no significant differences in the above indices between the VEGF group and the serum control group (all P > 0.05). Compared with the serum group, the middle- and high-dose serum containing Huangqi decoction groups had significantly inhibited proliferation, migration, and angiogenesis of rat LSECs induced by VEGF (all P < 0.05), and Western Blot showed significant reductions in the expression levels of CD31, ET-1, eNOS, and AKT/mTOR signaling pathway-related proteins (all P < 0.01). Conclusion A relatively high dose of serum containing Huangqi decoction can significantly inhibit the proliferation, migration, and tubulogenesis of rat LSECs induced by VEGF, possibly by regulating the AKT/mTOR signaling pathway.

Objective:To investigate the role of CD40/CD40L Pathway in the formation of silicosis fibrosis.Methods:Totally 64 inpatients were recruited and assigned to the silicosis group and the control group, 23 in each group. The alveolar lavage fluid was collected from all patients and isolated. The expression of CD40L protein was detected by Flow Cytometry. The level of IL-8、The IL-6、INF-γ and MCP-1 was detected by ELISA. Two groups of BALF were co-cultured with HFL-1 cells, the expression of Collagen I and α-SMA was detected by Immunohistochemistry.Results:Compared with the control group, CD40L was highly expressed on T lymphocyte cells in silicosis group ( P<0.05) , and the contents of IL-8、The IL-6、INF-γand MCP-1 in Silicosis group were significantly higher than those in control group ( P<0.05) . After co-culture of BALF and HFL-1 cells, the expression levels of Collagen I and α-SMA in Silicosis group were significantly higher than those in control group ( P<0.05) . Conclusion:CD40-CD40L cross-linking system can promote the activation of T cells, release inflammatory factors, promote the synthesis of collagen I and α-SMA by fibroblasts, make the lung fibrous tissue proliferate, and lead to the formation of silicosis fibrosis.

Objective:To investigate the role of CD40/CD40L Pathway in the formation of silicosis fibrosis.Methods:Totally 64 inpatients were recruited and assigned to the silicosis group and the control group, 23 in each group. The alveolar lavage fluid was collected from all patients and isolated. The expression of CD40L protein was detected by Flow Cytometry. The level of IL-8、The IL-6、INF-γ and MCP-1 was detected by ELISA. Two groups of BALF were co-cultured with HFL-1 cells, the expression of Collagen I and α-SMA was detected by Immunohistochemistry.Results:Compared with the control group, CD40L was highly expressed on T lymphocyte cells in silicosis group ( P<0.05) , and the contents of IL-8、The IL-6、INF-γand MCP-1 in Silicosis group were significantly higher than those in control group ( P<0.05) . After co-culture of BALF and HFL-1 cells, the expression levels of Collagen I and α-SMA in Silicosis group were significantly higher than those in control group ( P<0.05) . Conclusion:CD40-CD40L cross-linking system can promote the activation of T cells, release inflammatory factors, promote the synthesis of collagen I and α-SMA by fibroblasts, make the lung fibrous tissue proliferate, and lead to the formation of silicosis fibrosis.

OBJECTIVE: To investigate the efficacy of brain-targeted rapamycin (T-Rap) in treatment of epilepsy in rats. METHODS: Rapamycin nanoparticles targeting brain were prepared. The epilepsy model was induced by injection of pilocarpine in rats. The rats with pilocarpine-induced epilepsy were treated with rapamycin (Rap group) or brain-targeted rapamycin (T-Rap group). Seizure activity was observed by electroencephalography; the effect on mTOR signaling pathway was detected by Western blot; neuronal death and moss fiber sprouting were analyzed by Fluoro-Jade B (FJB) and Timm's staining, respectively. RESULTS: Electroencephalography showed that both preparation of rapamycin significantly reduced the frequency of spontaneous seizures in rats, and the effect of T-Rap was stronger than that of conventional rapamycin (<0.05). Western blot showed that the phosphorylation levels of S6K and S6 in T-Rap group were lower than those in Rap group (all <0.05), indicating that T-Rap had a stronger inhibitory effect on mTOR signaling pathway. FJB staining showed that T-Rap significantly decreased neuronal death, but there was no significant difference as compared with Rap group. Timm's staining showed that both preparations of rapamycin significantly reduced the germination of mossy fibers, while the effect of T-Rap was more pronounced than Rap group (<0.05). The inhibition of body weight gain of T-Rap group was less than that of Rap group (<0.05). CONCLUSIONS T-Rap has a better therapeutic effect on epilepsy than conventional rapamycin with a less adverse effects in rats.
Animals ; Brain ; drug effects ; Disease Models, Animal ; Epilepsy ; chemically induced ; drug therapy ; Neurons ; drug effects ; Pilocarpine ; Rats ; Rats, Sprague-Dawley ; Signal Transduction ; drug effects ; Sirolimus ; pharmacology ; therapeutic use ; Treatment Outcome

Accumulating data have revealed that abnormal activity of the mTOR (mammalian target of rapamycin) pathway plays an important role in epileptogenesis triggered by various factors. We previously reported that pretreatment with perifosine, an inhibitor of Akt (also called protein kinase B), abolishes the rapamycin-induced paradoxical increase of S6 phosphorylation in a rat model induced by kainic acid (KA). Since Akt is an upstream target in the mTOR signaling pathway, we set out to determine whether perifosine has a preventive effect on epileptogenesis. Here, we explored the effect of perifosine on the model of temporal epilepsy induced by KA in rats and found that pretreatment with perifosine had no effect on the severity or duration of the KA-induced status epilepticus. However, perifosine almost completely inhibited the activation of p-Akt and p-S6 both acutely and chronically following the KA-induced status epilepticus. Perifosine pretreatment suppressed the KA-induced neuronal death and mossy fiber sprouting. The frequency of spontaneous seizures was markedly decreased in rats pretreated with perifosine. Accordingly, rats pretreated with perifosine showed mild impairment in cognitive functions. Collectively, this study provides novel evidence in a KA seizure model that perifosine may be a potential drug for use in anti-epileptogenic therapy.
Animals ; Anticonvulsants ; pharmacology ; Brain ; drug effects ; pathology ; Convulsants ; toxicity ; Disease Models, Animal ; Epilepsy, Temporal Lobe ; chemically induced ; pathology ; Kainic Acid ; toxicity ; Male ; Neurons ; drug effects ; pathology ; Phosphorylcholine ; analogs & derivatives ; pharmacology ; Protein Kinase Inhibitors ; pharmacology ; Proto-Oncogene Proteins c-akt ; antagonists & inhibitors ; Rats ; Rats, Sprague-Dawley ; Status Epilepticus ; chemically induced ; pathology

Objective To investigate the mechanism of CD34+ progenitor cell differentiation in rat by observing the change relations between the eosinophils (EOS) and the content of Eotaxin and IL-5 in blood and the CD34+/CCR3+,CD34+/IL-5Rα+ in bone marrow after occupational asthma (OA) model rats are simulated,and to observe the effect of WTKYD Trraitioual Chinese Medicine intervention.Methods A total of 40 healthy male SD model rats (200 ～ 250 g weight) were randomly divided into model contrast Group,prednisone acetate intervention Group,WTKYD+1/2 prednisone acetate intervention Group and WTKYD intervention Group,10 in each group,and set a Group for blank contrast.Give them saline (20 ml/kg),prednisone acetate (8.22 mg/kg),WTKYD (20g/kg) +1/2 prednisone acetate (4.11 mg/kg) and WTKYD (20 g/kg) intervention respectively.By means of cell count,immunohistochemical,ELISA,flow cytometry technique,situ hybridization and so on,to observe EOS anti the expression of Eotaxin in lung tissue,the EOS in peripheral blood,the content of Eotaxin and IL-5 in blood as well as the expression of CD34+/CCR3 + and CD34+/IL-5Ra+ in bone marrow respectively.Results The number of EOS,the content of Eotaxin and IL-5,the expression of CD34+/CCR3+ and CD34+/IL-5Ra+ in Model Contrast Group were higher in Blank Contrast Group,the difference was statistically significant (P＜0.01),while they were lower in mnedical intervention Groups when comparing to Model Contrast Group,the difference was statistically significant (P＜0.01 or P＜0.05),and the above items in WTKYD +1/2 Prednisone Acetate Intervention Group were even lower thau in Prednisone Acetate Intervention Group and WTKYD Intervention Group,the difference was statistically significant (P＜0.05).EOS in lung tissue is highly positive related to the content of Eotaxin and IL-5 in peripheral blood as well as the expression of CD34+/CCR3 and CD34+/IL-5Rα in bone marrow (0.9666,0.9829,0.9142,0.8874).Conclusion The increase of internnd EOS in lung tissue is related to the up-regulated expression of CD34+/CCR3+ and CD34+/IL-5Ra+ in bone marrow after antigens in Occupational Asthma model rats are stimulated.Through down-regulating it's expression to restrain the differentiation of CD34 + progenitor cells towards EOS,meanwhile,the collaboration of WTKYD and prednisone acetate possess a certain synergistic action.

Objective To investigate the mechanism of CD34+ progenitor cell differentiation in rat by observing the change relations between the eosinophils (EOS) and the content of Eotaxin and IL-5 in blood and the CD34+/CCR3+,CD34+/IL-5Rα+ in bone marrow after occupational asthma (OA) model rats are simulated,and to observe the effect of WTKYD Trraitioual Chinese Medicine intervention.Methods A total of 40 healthy male SD model rats (200 ～ 250 g weight) were randomly divided into model contrast Group,prednisone acetate intervention Group,WTKYD+1/2 prednisone acetate intervention Group and WTKYD intervention Group,10 in each group,and set a Group for blank contrast.Give them saline (20 ml/kg),prednisone acetate (8.22 mg/kg),WTKYD (20g/kg) +1/2 prednisone acetate (4.11 mg/kg) and WTKYD (20 g/kg) intervention respectively.By means of cell count,immunohistochemical,ELISA,flow cytometry technique,situ hybridization and so on,to observe EOS anti the expression of Eotaxin in lung tissue,the EOS in peripheral blood,the content of Eotaxin and IL-5 in blood as well as the expression of CD34+/CCR3 + and CD34+/IL-5Ra+ in bone marrow respectively.Results The number of EOS,the content of Eotaxin and IL-5,the expression of CD34+/CCR3+ and CD34+/IL-5Ra+ in Model Contrast Group were higher in Blank Contrast Group,the difference was statistically significant (P＜0.01),while they were lower in mnedical intervention Groups when comparing to Model Contrast Group,the difference was statistically significant (P＜0.01 or P＜0.05),and the above items in WTKYD +1/2 Prednisone Acetate Intervention Group were even lower thau in Prednisone Acetate Intervention Group and WTKYD Intervention Group,the difference was statistically significant (P＜0.05).EOS in lung tissue is highly positive related to the content of Eotaxin and IL-5 in peripheral blood as well as the expression of CD34+/CCR3 and CD34+/IL-5Rα in bone marrow (0.9666,0.9829,0.9142,0.8874).Conclusion The increase of internnd EOS in lung tissue is related to the up-regulated expression of CD34+/CCR3+ and CD34+/IL-5Ra+ in bone marrow after antigens in Occupational Asthma model rats are stimulated.Through down-regulating it's expression to restrain the differentiation of CD34 + progenitor cells towards EOS,meanwhile,the collaboration of WTKYD and prednisone acetate possess a certain synergistic action.

OBJECTIVETo determine the effects of ginsenosides Rb1(GSRb1) on learning and memory and expression of somatostatin (SS) in the hippocampus and the frontal cortex in rat model of sleep deprivation (SD).

METHODSRats were randomized into groups of SD 2 d, SD 4 d, SD 6 d, and SD 0 d, while each group was sub-divided into GSRb1 group and normal saline (NS) sub-groups. Rats were intraperitoneal administered with 30 mg/(kg*d) of GSRb1 or NS for 7 d, then the learning and memory abilities were examined by measuring average swimming speed and mean escape latency using Morris maze.Expression of somatostatin was detected with immunohistochemical method and image analysis in the hippocampus and the frontal cortex.

RESULTSCompared with SD 0 d rats, SD rats exhibited significant decrease in the average swimming speed and increase in the escape latency (P <0.01). The expression of somatostatin in the hippocampus was decreased significantly in SD 2 d, SD 4 d and SD 6 d rats (P<0.05). However, decrease was only observed in SD 4 d and SD 6 d rats in the frontal cortex (P <0.05). Parallel comparison between NS control and GSRb1 treated rats demonstrated that rats treated with GSRb1 in each subgroup exhibited faster swimming speed and shorter escape latency (P <0.05). Meanwhile, the expression of somatostatin was increased in SD 2 d, SD 4 d and SD 6 d rats in the hippocampus and in SD 4 d and SD 6 d rats in the frontal cortex (P <0.05), respectively.

CONCLUSIONGSRb1 enhances the expression of somatostatin in sleep deprivation rats and subsequently may improve learning and memory abilities of rats.

Animals ; Brain ; metabolism ; Disease Models, Animal ; Ginsenosides ; pharmacology ; Learning ; drug effects ; Male ; Memory ; drug effects ; Rats ; Rats, Sprague-Dawley ; Sleep Deprivation ; metabolism ; Somatostatin ; metabolism

The chiral separation of three triazole pesticides, i.e. diniconazole, triadimefon and triadimenol was studied on a Chiralcel OJ-H and a Chiralcel OD-H HPLC chiral columns. The optical rotation quality of diniconazole and triadimefon enantiomers was measured and the absolute configurations of individual enan-)tiomers) were further concluded. On this basis, the absolute configurations of the four triadimenol stereoisomers were deduced via the reductive experiment of triadimefon to triadimenol. Furthermore, the chiral stability of the three triazole pesticides in organic solvents and buffer solutions was investigated. The results showed the obvious enantiomerization was observed as for triadimefon in methanol, ethanol and water, whereas dinicona-)zole) and triadimefon were chiral stable in organic solvents and water. The enantiomerization of triadimefon would be accelerated at higher temperature and in alkaline media.

Objective To screen and isolate the radioresistance related genes of IRM-2 mice.Methods cDNA library of IRM-2 mice was constructed by SMART technique.Total RNA was isolated from spleens of IRM-2 male mice.The first-strand cDNA was synthesized by using PowerScript reverse transeriptase,and double-strand cDNA was synthesized and amplified by long PCR.The PCR products were purified,digested with restriction enzyme Sfi I.The ds-cDNA fragment lessthan 500 bp was fractionated and ligated to the Sfi I-digested pDNR-LIB vector.The ligation mixture was transformed into E.coil DH5α by electroporution transformation to generate the unamplified cDNA library.The quality of cDNA library was identified by PCR technique.130 clones from cDNA library were sequenced and compared with GenBank database.Results The cDNA library contained 2.25 x 106 independent clones with an average insert size of 1.2 kb.The ratio of recombination and full-length was 95% and 55%,respectively.21 pieces of EST sequences from cDNA library were not the same as the known mice genes and registered into GenBank EST database,with registered number DW474856-DW474876.Conclusions cDNA library of IRM-2 mice has been constructed successfully.21 pieces of EST implies that radioresistance correlative genes may be in IRM-2 mice,which will lay a foundation for isolating and identifying radioresistance related genes in further study.