Objective To observe the changes of intracellular free calcium and the expression of CaM in the hippocampal neurons of posttraumatic stress disorder (PTSD) rats and to further investigate the neurobiological mechanisms. Methods The SPS-method was used to set up the rat PTSD models. A total of sixty male Wistar rats were randomly divided into 12 hours,1 day,4 days,7 days groups of SPS and normal control group. The intracellular free calcium was examined by fluorescence spectrophotometer. The expression of CaM was detected by using immunohistochemistry, Western blotting and RT-PCR. Results The intracellular free calcium level in the hippocampus of experimental rats was markedly increased 12 hours after SPS stimulation,and reached the peak after 1 day, then gradually decreased to normal level after 7 days. The expression of CaM in the hippocampus 1day after SPS was also the highest and then gradually decreased.Conclusion The lasting dysfunction of Ca~(2+)/CaM signaling cascades in hippocampal may play important roles in the pathogenesis of PTSD rats.

By molecular cloning technique, the expressing plasmids pDOR-SV40-Bcl-2 cDNA were successfully constructed. The reconstructed plasmids with lipofectamine were transduced into gastric cancer cell line SGC7901 and then the positive clones which contained the reconstructed plasmids were choosed by G418. Circa 150 positive clones were choosed from 2 x 10_5 gene transduced cells, which suggested that the transducing efficiency was more than 1%o; 2 positive clones were expanded and passage, then 1 drug-resistance cell strain (SGC7901 anBcl-2 cells) was obtained. The bloting results suggested, Bcl-2 cDNA were expressed in both gene transduced and not transduced cell strains, but the expressing level of mRNA and protein in gene transduced cell strain was very low than that in gene not transduced cell strain were positive by the means of Southern blot, Northern blot and Western blot. This results showed that normally in gastric cancer cells the expressing level of Bcl-2 gene was very high, and the cDNA fragment of antisense Bcl-2 were successfully transduced into gastric cancer cells, and in gene transduced cell strains the expressing of Bcl-2 gene was effi-cientlv blocked.

Objective:To investigate YKL-40-mediated inflammation in human bronchial epithelial cells and analyzed the soluble factors secreted by bronchial epithelial cells exposed to YKL-40 that were responsible for increasing proliferation and migration of primary normal human bronchial smooth muscle cells (BSMCs).Methods:YKL-40-induced inflammation was assayed in two human bronchial epithelial cells (BEAS-2B cell line and primary human bronchial epithelial cells ,namely HBECs).In addition,we treated BEAS-2B cells and HBECs with YKL-40,and added the conditioned culture media ( YKL-40-BEAS-2B-CM) and ( YKL-40-HBECs-CM) to BSMCs.The proliferation and migration of BSMCs were determined by premixed WST-1 cell proliferation reagent and QCM chemotaxis migration assay ,respectively.Results: Bronchial epithelial cells treated with YKL-40 resulted in a significant increase of IL-8 production,but have no effect about RANTES ,Eotaxin and TNF-α.YKL-40-BEAS-2B-CM and YKL-40-HBECs-CM induced IL-8 was found to further stimulate proliferation and migration of BSMCs ,and the effects were inhibited after neutralizing IL-8.Conclusion:Through investigating the interaction of airway epithelium and smooth muscle ,our findings implicate that YKL-40 may be involved in the inflammation of asthma by induction of IL-8 from epithelium,subsequently contributing to BSMCs proliferation and migration.Moreover, inhibition of IL-8 signaling is a potential therapeutic target for YKL-40-induced inflammation and remodeling of asthma.

Objective: To further understand the role of folic acid supplements rivaling MTHFR gene silencing in pathogenesis of NCLP, RNA interference (RNAi) was applied to knock down MTHFR in mouse embryonic palatal mesenchymal (EPM) cells. Methods: MTHFR ShRNA expression vector were transfected into the primary cultured EPM cells. MTT was used to observe cell proliferation after MTHFR gene silencing. FCM was used to observe cell cycle after MTHFR gene silencing. Results: The results showed the cells proliferation had an inequality amelioration after using folic acid supplements in MEPM cells with MTHFR gene silencing. Using folic acid supplements rivaled the effect of MTHFR gene silencing had a dose-dependent manner. Using 20 μg/ml folic acid supplements could improve the cell proliferation to achieve normal level of cell proliferation. Conclusion: MTHFR gene is an important candidate gene of NCL/P. Using folic acid supplements could prevent teratogenic MTHFR gene silencing for embryonic palate development.

Data mining, as known as knowledge discovery in databases, is a non-trivial process of revealing the implied, previously unknown and potentially useful information from the massive data. In recently years, the applications of data mining in the field of pharmaceutical research of traditional Chinese medicine have widespread. Especially in the field of the heritage of experiences of na-tional medical masters, data mining plays an important role. In this study, we would expound of the use of methods of data mining in the heritage of experiences of national medical masters, and analyze their advantages and disadvantages, such as association rules, Bayesian networks, neural networks, and decision trees.
Data Mining ; Databases, Pharmaceutical ; Medicine, Chinese Traditional ; Neural Networks (Computer)

OBJECTIVETo observe application value of multispectral animal living imaging technology in rats model of osteoarthritis.

METHODSFifteen male SD rats weighed (180 +/- 20) g (3 months old) were received intra-articular injection of iodoacetic acid for establishing osteoarthritis. Articular cavity of left knee of rats were injected into 50 microl iodoacetic acid. The same volume of sterile saline was injected into right knee articular cavity as control. X-ray living imaging and bone mineral density were observed at 2 and 4 weeks after establishment of model. After 4 weeks,rats were sacrificed and their bilateral joints were collected and determined histologically based on Collins classification and Kellgren-Lawrence classification.

RESULTSOsteoarthritis model was successfully established, compared with control group, model group showed typical manifestation of osteoarthritis, including irregular cartilage surface,osteophyte formation,joint deformity and cartilage defect,and combined with significant decrease of bone density (P < 0.01), while the decrease was not obvious in proximal tibia (P < 0.05). After 2 weeks, knee joints in model group was classified as Collins grade 1 and Kellgren-Lawrence grade 2,then classified as Collins grade 4 and Kellgren-Lawrence grade 3 after 4 weeks,control group showed smooth articular surface,normal joint space and intact cartilage surface, knee joints was classified as Collins and Kellgren-Lawrence grade 0, and bone density of distal femur and proximal tibia were normal.

CONCLUSIONMultispectral animal living imaging technology could be used in dynamic observation of living imaging and detection of bone density in the animal model of osteoarthritis, and it is significant for evaluation of osteoarthritis model, and its realted tesearch.

Animals ; Bone Density ; Disease Models, Animal ; Humans ; Knee Joint ; diagnostic imaging ; Male ; Osteoarthritis ; diagnosis ; diagnostic imaging ; physiopathology ; Radiography ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo observe the effect of Shen warming Pi strengthening method on expressions of serum T cell subsets (C045+%, C03+%, and C04 +/COB+) in diarrhea-predominant irritable bowel syndrome (IBS-0) rats. Methods An IBS-0 rat model was established referring to AL-Chaer's modeling method combined with tail clamp and intragastric administration of sanna leaf. After modeling 30 SO rats were randomly divided into 6 groups according to random digit table, i.e., the model group, the high, middle, low dose Wenshen Jianpi Recipe (WJR) groups, and the Sishen Pill control group, 6 in each group. A normal control group consisting of 6 SO rats were also set up. Rats in high, middle, low dose WJ R groups were administered by gastrogavage with boil-free WJ R at the daily dose of 3. 100, 1. 550, 0. 775 g/kg, respectively. Rats in the Sis hen Pill control group were administered by gastrogavage with boil-free Sis hen Pill at the daily dose of 0. 736 g/kg. Equal volume of normal saline was given by gastrogavage to rats in the model group and the normal control group. All medication lasted for 2 successive weeks. Rats' general state, expressions of T cell subsets (CD45+%, CD3+%, and CD4+ /CDB+) changes were observed.

RESULTSCompared with the normal control group, expressions of CD45+% and CD3+% increased, but CD4+ /CDB+ decreased with statistical difference (P < 0. 05). Compared with the model group, expressions of CD45+% and CD3+% decreased, but CD4+ ICDB+ increased with statistical difference in high, middle, low dose WJR groups, and the Sis hen Pill control group (P <0. 05). Compared with the Sis hen Pill control group, there was statistical difference in all indices except CD45+ value in the low dose SWPSM group (P <0. 05). Compared with the low dose WJ R group, the expression of CD3+% decreased in high and middle dose WJR groups, and the Sis hen Pill control group; CD4+ /CD8+ increased in the Sishen Pill control group and the high dose SWPSM group (all P < 0. 05).

CONCLUSIONSWJR showed better treatment effect. The mechanism of Shen warming Pi strengthening method might be achieved by regulating expressions of CD45+% and CD3+%, and CD4+ /CD8+ ratios.

Animals ; Drugs, Chinese Herbal ; Female ; Irritable Bowel Syndrome ; therapy ; Leukocyte Common Antigens ; metabolism ; Medicine, Chinese Traditional ; Rats ; T-Lymphocyte Subsets ; metabolism

Objective To investigate the expression and possible mechanism of miR-21 and Ski-related novel protein N( SnoN) in the renal fibrosis diabetic process.Methods The animal model was established by tail-vein injection of Streptozotocin,and the other group were normal control ( NC) group.After 10 weeks, the rats were sacrificed to measure biochemical parameters and renal index , and to observe the changes of pathomorphology by HE staining as well.Meanwhile, immunohistochemistry and Western blot were employed to examine protein ex-pression of E-cadherin,α-smooth muscle actin(α-SMA), fibronectin(FN), collagen-Ⅰ(Col-Ⅰ), collagen-Ⅲ(Col-Ⅲ), transforming growth factor-β1(TGF-β1), Smad3, p-Smad3(Ser423/425) and SnoN in the renal tissue. In addition, the expression of SonN mRNA and miR-21 were detected by qPCR.Results In DM group,the ex-pressions of Col-Ⅰ, Col-Ⅲ and FN in renal interstitium were increased ( P <0.05 ) , TGF-β1 increased (P<0.05),while E-cadherin decreased(P<0.05).Compared with NC group, the expression of α-SMA,p-Smad3 (Ser423/425) protein increased in DM group(P<0.05),while the protein level of SnoN decreased but the level of SnoN mRNA increased ( P <0.05 ) .Moreover, the level of miR-21 markedly increased in DM group ( P <0.05 ) .Conclusions TGF-β1 may up-regulate the expression of miR-21 but restrain the translational expression of SnoN, aggravating fibrosis.

AIM: To investigate the effects of bone morphogenetic protein 7 ( BMP-7 ) on the expression of transcription factor E2A and inhibitor of differentiation 2 (Id2) in the renal tubule epithelial cells(NRK-52E)exposed to high glucose, and to explore its possible mechanism of improving renal tubular fibrosis induced by high glucose.METH-ODS:The NRK-52E cells were divided into control group, high glucose (HG) group and high glucose with different doses of BMP-7 (10μg/L and 20μg/L) group.The cells in HG group and BMP-7 group were cultured for 12 h, 24 h and 48 h. The protein expression of Id2, E2A, E-cadherin,α-smooth muscle actin (α-SMA) and collagen-I was detected by Western blot.In addition, the mRNA expression of Id2 was detected by real-time PCR.RESULTS:Compared with control group, the mRNA and protein levels of Id2 and the protein level of E-cadherin were down-regulated, while the protein levels of E2A,α-SMA and collagen-I were up-regulated in HG group (P<0.05).Compared with HG group, the mRNA and pro-tein levels of Id2 and the protein level of E-cadherin were significantly up-regulated, while the protein expression of E2A,α-SMA and collagen-I was significantly down-regulated in 20 μg/L BMP-7 group ( P<0.05 ) .The correlation analysis showed that the Id2 protein level was negatively correlated with the E2A protein level (P<0.05).CONCLUSION:BMP-7 may intercept the process of renal tubule fibrosis induced by high glucose via promoting the expression of Id2 and inhibi-ting the expression of E2A at protein level.

OBJECTIVETo observe the clinical effect of low-dose once-daily tadalafil combined with Shuganyiyang Capsules in the treatment of mild-to-moderate erectile dysfunction (ED).

METHODSNinety patients with mild-to-moderate ED were equally randomized to groups A, B and C to receive Shuganyiyang Capsules, tadalafil, and tadalafil + Shuganyiyang Capsules, respectively. The scores of the patients on IIEF-5 and SF-PAIRS (15-Item Short Form of Psychological Interpersonal Relationship Scales) were recorded before and at 1 and 3 months after treatment.

RESULTSThe IIEF-5 scores of groups A, B and C were 10.13 +/- 1.55, 11.00 + 1.60 and 10.73 +/- 1.91 before treatment, and 13.77 +/- 2.11, 17.77 +/- 2.13 and 17.17 +/- 3.84 at 1 month after treatment, significantly higher in B and C than in A (P <0. 001) , but with no remarkable difference between B and C (P =0. 411). At 3 months after treatment, the IIEF-5 scores were 15.77 +/- 2.05, 18.07 +/- 2.24 and 19.37 +/- 3.76 in the three groups, dramatically higher in B and C than in A (P <0.001) as well as in C than in B (P<0.05). The scores on sexual self-confidence, sexual spontaneity and time concerns in SF-PAIRS were 3.90 +/-0.80, 8.67 +/- 1.94 and 14.43 +/- 1.92 before medication, 5.83 +/- 1.02, 9.90 +/- 1.75 and 11.17 +/- 1.68 at 1 month and 6.73 +/- 0.98, 11.07 +/- 2.08 and 10.67 +/-1.60 at 3 months after medication in group A; 4.17 +/- 0.87, 9.37 +/-1.43 and 14.47 +/-1.57 before medication, 6.47 +/-0.78, 10.83 +/- 2.18 and 10.20 +/-1.56 at 1 month and 6.83 +/-0.91, 11.30 +/- 1.88 and 9.47 +/- 1.57 at 3 months in group B; and 4.23 +/-0. 94, 9.50 +/- 1.89 and 14.67 +/- 2.91 before medication, 8.03 +/- 1.67, 13.43 +/-1.10 and 9.70 +/-1.21 at 1 month and 8.93 +/- 1.78, 14.70 +/- 1.26 and 8. 87 +/- 0. 97 at 3 months in group C. Compared with the baseline, the SF-PAIRS scores of the three groups were all significantly improved after treatment (P <0. 05) , and markedly higher in C than in the other two groups (P <0.05).

CONCLUSIONLow-dose once-daily tadalafil combined with Shuganyiyang Capsules is obviously effective in the treatment of mild-to-moderate ED, which not only improves the patients'erectile function, sexual self-confidence and sexual spontaneity, but also reduces their time concerns.

Adult ; Carbolines ; administration & dosage ; therapeutic use ; Drugs, Chinese Herbal ; therapeutic use ; Erectile Dysfunction ; drug therapy ; Humans ; Male ; Tadalafil ; Treatment Outcome