OBJECTIVEEffects of ginsenoside Rb1, Rg1 and Re on neurotrophic factor signal transduction pathway using liposome-mediated transfection of eukaryotic cells approach.

METHODThe injury model was established by treating SH-SY5Y cells with 0.6 mmol x L(-1) of corticosterone (CORT) by 24 h. SH-SY5Y cell were pretreated with CORT for 30 min followed by co-treated with 120,60 and 20 micromol x L(-1) of Rb1, 120, 80 and 40 micromol x L(-1) of Rg1 and 120, 80 and 40 micromol x L(-1) of Re for 24 h. Cells viability was determined by Cell Counting Kit (CCK) assay. CREB expressing Luciferase reporter gene was constructed and transfected with plasmid containing hRaf, hcAMP, hAkt, hCaMK gene into human embryonic kidney (HEK293) cells using liposornal transfection reagent lipofection 2000. The expression of CREB before and after it addion of Rb1, Rg1 and Re was examined by Luc assay system and Western blotting.

RESULTCompared with normal control group, CORT significantly decreased the viability of SH-SY5Y cells to 67.21% (P < 0.01). CCK results show that Rb1 (60 micromol x L(-1)), Rg1 (80 micromol x L(-1)) and Re (80 micromol x L(-1)) on SH-SY5Y cells have significant protective effect (P < 0.01). Lucassay and Western blotting results show that the gene and protein levels of CREB increased significantly through the pathway of Raf and Akt with Rb1 and Rg1 (P < 0.01), Re can increase significantly the gene and protein levels of CREB through the pathway of Raf and CaMK II.

CONCLUSIONRb1, Rg1 and Re protects SH-SY5Y cells from CORT-induced damage and the neuroprotective mechanism may be associated with the Raf-CREB, Akt-CREB and CaMK II -CREB pathways.

Calcium-Calmodulin-Dependent Protein Kinase Type 2 ; genetics ; metabolism ; Cell Line ; Cell Survival ; drug effects ; Cyclic AMP Response Element-Binding Protein ; genetics ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; Genes, Reporter ; Ginsenosides ; pharmacology ; Humans ; Panax ; chemistry ; Proto-Oncogene Proteins c-akt ; genetics ; metabolism ; Signal Transduction ; drug effects ; raf Kinases ; genetics ; metabolism

Five compounds (tenuifoliside C, tenuifoliside D, telephiose A, telephiose C and polygalaxanthone III) from polygala tenuifolia wild were incubated together with CYP probe substrate in human liver microsomes to investigate the inhibitory effect towards CYP450 enzyme. Phenacetin (CYP1A2), coumarin (CYP2A6), paclitaxel (CYP2C8), diclofenac (CYP2C9), S-mepheriytoin (CYP2C19), dextromethorphan (CYP2D6), chlorzoxazone (CYP2E1), midazolam (CYP3A) were selected as the isoforfn specific substrate. And the formation of paracetamol, 7-hydroxycoumarin, 6alpha-hydroxy paclitaxel, 4'-hydroxydiclofenac, dextrorphan, 6-hydroxychlorzoxazone, 1'-hydroxymidazolam, 4'-hydroxymephenytoin were detected respectively to measure the effect towards CYP450 by high-pressure liquid chromatography (HPLC). The result shows that five compounds from polygala tenuifolia willd significantly inhibit chlorzoxazone 6-hydroxylation catalyzed by CYP2E1, while showed no effect towards CYP1A2, CYP2A6, CYP2C8, CYP2C9, CYP2C19, CYP2D6, CYP3A. And IC50 value was 38.73, 54.14, 61.77, 62.22, 50.56 micromol x L(-1), respectively.
Cytochrome P-450 Enzyme System ; metabolism ; Esters ; pharmacology ; Glycosides ; pharmacology ; Humans ; Microsomes, Liver ; drug effects ; enzymology ; Oligosaccharides ; pharmacology ; Polygala ; chemistry ; Xanthones ; pharmacology

Objective To study the mutations of Env sequence of SIVmac239 after infection of Chinese rhesus monkeys, and compare the differences and characteristics of Gp120 sequences of enterotropic and neurotropic SIV strains. Methods Six strains of simian immunodeficiency virus were analyzed in this study: four separated from peripheral blood mononuclear cells of SIVmac239-infected monkeys and two neurotropic SIVmac251 strains.Isolated and cultured monoclonal virus was obtained by limiting dilution assay.Gp120 sequences were amplified after the RNA extraction and phylogenetic analysis was processed thereafter.So did the Gp120 amino acid sequence and N-glycosylation sites analysis of the enterotropic and neurotropic strains.Results SIVmac239 had different mutations in four rhesus monkeys.The diversity in amino acid sequences of the enterotropic and neurotropic strains concentrated in the V1 and V4 regions of Gp120.The enterotropic strains had an addition of glycosylation site in V4 but the glycosylation site changes of neurotropic strains were located in the conservative regions of C1, C2 and C3.Conclusions The addition of one glycosylation site in V4 region of GP120 and loss of one glycosylation site in C1 region are associated with enhanced enterotropism and neurotropism.The differences between the enterotropic and neurotropic strains are not dipicted in Gp120 V3 region which is closely related with the tropism of strains.

Objective To study the effect of repeated rectal exposure of low -dose simian immunodeficiency virus on the systemic cellular immunity in monkeys .Methods Eight 3-to 4-year old rhesus macaques ( Macaca mulatta) (male:female 1:1) were used in this study.The monkeys were inoculated with 10 TCID50 SIVmac239 virus through rectum twice a week for consecutive 6 weeks to establish a multiple rectal exposure model of SIVmac 239 virus infection.Then, plasma viral load, CD4+ T cell count, T cell subsets and IFN-γsecretion of the experiment monkeys were determined . Results Low-dose SIVmac239 virus induced some changes in the immune system through the rectal mucosa , but didn’t induce typical infection.Repeated rectal mucosal low-dose virus exposure can activate the cellular immune system . Conclusions This study defines the effect of repeated low -dose simian immunodeficiency virus exposure on the systemic cellular immunity, and provided basic information for HIV-1 vaccine research.

The objective of this study was to investigate the immunophenotype of T-lineage acute lymphoid leukemia (T-ALL) and to find valuable immunologic markers in T-ALL diagnosis and therapy. Four-color multiparametric flow cytometry(FCM) with CD45/SSC gating was used for immunophenotyping of 95 patients with newly diagnosed T-ALL. The results demonstrated that T-ALL occurred more frequently in males younger than 30 years of age and was usually accompanied by a high WBC count and tumor mass at diagnosis. Univariate analysis showed an influence on achievement of CR1 for age (< 30 years) but not for WBC count and tumor mass. According to WHO (2008) classification of tumors of haematopoietic and lymphoid tissues, 87 patients with confirmed subtype included 27 cases of Pro-T-ALL (31.0%), 31 cases of Pre-T-ALL (35.6%), 23 cases of cortical-T-ALL (26.4%), 6 cases of medullary-T-ALL (6.9%). CD34 expression in Pro-T-ALL was significantly higher than that of Pre-T-ALL (p = 0.001). After the first chemotherapy, the complete remission rate in Pro-T-ALL was statistically lower than that of Pre-T-ALL. Besides, the complete remission rate of immature T-ALL (including Pro-T-ALL and Pre-T-ALL) was also significantly lower than that in mature T-ALL (including cortical-T-ALL and medullary-T-ALL). Myeloid antigen (CD13, CD33) expression was associated with T-ALL subtype and treatment effect. While 66.7% of CD13(+) patients belonged to Pre-T-ALL, most (60.0%) of CD33(+) patients were classified into Pro-T-ALL; CD13 expression had no effect on CR1 rate whereas CD33(+) patients had worse treatment effect compared with CD33(-) groups (p = 0.001). Notably, the expression of CD117 reached up to 26.7% and the positive cases were primarily distributed in pro-T-TAll and pre-T-ALL. It is found that CD117 expression in CD34(-) group was homogeneous and CD117 expression level was less than 10% in 73.2% patients, but CD117 expression level in CD34(+) group was not homogenous, in which group the CD117 expression levels < 10%, 10% - 20% and > 20% were 44.2%, 17.3% and 38.5% respectively. As compared with CD34(-) group, the proportion of patients with CD117 expression levels < 10%, > 20% in CD34(+) group was higher, and there was significant difference between these 2 group. It is concluded that immunophenotype has great value in T-ALL diagnosis, classification as well as treatment. Flow cytometry provides access to find valuable immunologic markers for T-ALL biological research.
Adolescent ; Adult ; Aged ; Aged, 80 and over ; CD13 Antigens ; metabolism ; Child ; Child, Preschool ; Female ; Humans ; Immunophenotyping ; Infant ; Male ; Middle Aged ; Precursor T-Cell Lymphoblastic Leukemia-Lymphoma ; classification ; immunology ; therapy ; Proto-Oncogene Proteins c-kit ; metabolism ; Sialic Acid Binding Ig-like Lectin 3 ; metabolism ; Young Adult

OBJECTIVETo study the pathologic change and molecular regulation in cell proliferation and apoptosis of gastric mucosa in rats with chronic atrophic gastritis (CAG), and evaluate the possible mechanisms.

METHODSRats were administered with 60% alcohol or 2% salicylate sodium, 20 mmol/L deoxycholate sodium and 0.1% ammonia water to establish chronic atrophic gastritis (CAG) models. The gastric specimens were prepared for microscopic view with hematoxylin and eosin (H-E) and alcian blue (A-B) stain. The number of infiltrated inflammatory cells, the thickness of the mucosa gland layer (microm) and the number of gastric glands were calculated. The damage of barrier in mucosa with erosion or ulceration, and the thickness of mucin were examined by scanned electron microscope (SEM). The levels of PGE(2), EGF (epiderminal growth factor) and gastrin in the serum were measured with radioimmunoassay or ELISA method. The immunohistochemistry method was used to observe the number of G cells, the expression of protein of EGFR (EGF receptor), C-erbB-2, p53, p16 and bcl-2 in gastric tissue.

RESULTSUnder SEM observation, the gastric mucosa was diffused erosion or ulceration and the thickness of mucin was decreased. Compared with normal rats, the grade of inflammatory cell infiltration in CAG rats was elevated, whereas the thickness and number of gastric gland were significantly lower (P<0.05). Compared with normal level of (0.61+/-0.28) microg/L, EGF in CAG (2.24+/-0.83) microg/L was significantly higher (P<0.05). The levels of PGE(2) and gastrin in serum were significantly lower in CAG rats than that in normal rats (P<0.05). Immunohistochemistry detection showed that the number of G cell in antrum was lower in CAG group (P<0.05). Immuno-stain showed EGFR protein expression in the basal and bilateral membrane, and the cytoplasma in atrophic gastric gland, while negative expression was observed in normal gastric epithelial cells. Positive staining of p53 and p16 protein was localized in the nucleus of epithelial cells. The former was higher positively expressed in atrophic gland, while the later was higher positively stained in normal gastric tissue. bcl-2 protein was positively stained in the cytoplasma in atrophic gastric gland, while very weakly stained in normal gastric tissue.

CONCLUSIONThe pathological findings in gastric gland accorded with the Houston diagnostic criteria of antrum-predominant CAG. CAG in rats was related with the damage of barrier in gastric mucosa and the misbalance of cell proliferation and apoptosis. There was high protein expression of oncogene, while inhibitor of suppressor gene in CAG rats indicated high trend of carcinogenesis.

Animals ; Chronic Disease ; Epidermal Growth Factor ; blood ; Gastric Mucosa ; chemistry ; pathology ; Gastrins ; blood ; Gastritis, Atrophic ; metabolism ; pathology ; Immunohistochemistry ; Male ; Proto-Oncogene Proteins c-bcl-2 ; analysis ; Rats ; Rats, Sprague-Dawley ; Receptor, Epidermal Growth Factor ; analysis ; Tumor Suppressor Protein p53 ; analysis

The changes of microRNA expression in rat hippocampus after traumatic brain injury (TBI) were explored. Adult SD rats received a single controlled cortical impact injury, and the ipsilateral hippocampus was harvested for the subsequent microarray assay at three time points after TBI: 1st day, 3rd day and 5th day, respectively. We characterized the microRNA expression profile in rat hippocampus using the microRNA microarray analysis, and further verified microarray results of miR-142-3p and miR-221 using quantitative real-time PCR. Totally 205 microRNAs were identified and up-/down-regulated more than 1.5 times. There were significant changes in 17 microRNAs at all three time points post-TBI. The quantitative real-time PCR results of miR-142-3p and miR-221 indicated good consistency with the results of the microarray method. MicroRNAs altered at different time points post-TBI. MiR-142-3p and miR-221 may be used as potentially biological markers for TBI assessment in forensic practice.

Objective: In comparison with thrombus aspiration, to study the safety and effcacy of precise intracoronary retrograde thrombolysis during primary percutaneous coronary intervention (PCI) in patients with acute ST-segment elevation myocardial infarction (STEMI).
Methods: A total of 123 consecutive patients with acute STEMI received primary PCI in our hospital from 2014-01 to 2015-12 were enrolled.The patients were randomly divided into 2 groups: RT group, the patients received precise intracoronary retrograde thrombolysis (RT),n=60 and TA group, the patients received thrombus aspiration (TA),n=63, among them, 3 patients with failed TA were excluded. Primary end points included occurrence rates of no-lfow after PCI and ST-segment resolution (STR)≥50% at (60-90) min after PCI; primary safety end points included occurrence rates of in-hospital stroke and TIMI-hemorrhage events.
Results:①Compared with TA group, RT group showed decreased no-lfow rate after PCI (1.7% vs 15.0%),P=0.008 and increased rate of STR≥50% after PCI (65.0% vs 45.0%),P=0.028, improved LVEF by echocardiography (50.7±8.6) % vs (46.7±8.3)%,P=0.011. The in-hospital MACE occurrence rate was similar between 2 groups,P>0.05.②No in-hospital stroke or TIMI-hemorrhage events occurred in neither group.
Conclusion: Intracoronary retrograde precise thrombolysis had the similar safety to thrombus aspiration during primary PCI in patients with acute STEMI, it may reduce no-relfow rate and improve left ventricular function after PCI.

Objective: To compare the efifcacy of direct and pre-dilated atherectomy (RA) for treating the patients with calciifed coronary lesions. Methods: A total of 137 coronary artery disease (CAD) patients received RA treatment in our hospital from 2010-04 to 2014-09 were retrospectively studied. The ischemic related lesions were all deifned as calciifed coronary lesions. The patients were divided into 2 groups: Direct RA group,n=81 and Pre-dilated RA group, the patients received balloon dilatation followed by RA,n=56. The procedural features, complications, in-hospital and 1 year occurrence rates of major cardiaccerebral vascular events (MACCE) were compared between 2 groups. Results: Compared with Pre-dilated RA group, Direct RA group had the less pre-stent balloon application,P=0.000 and the higher maximum post-dilatationpressure,P=0.004; lower rate of in-operative complication (14.8% vs 32.1%),P=0.016; higher rate of acute lumen gain (128.52±75.77) % vs (77.12±27.01) %,P=0.004; lower MACCE occurrencerate(7.3% vs 23.6%) at 1 year period,P=0.006.Cox regression analysis presented that the following indexes were related to MACCE occurrence within 1 year of RA treatment: balloon dilatation before RA (HR=8.166, 95% CI 1.872-35.614,P=0.005), left main disease (HR=13.649, 95% CI 2.983-62.440,P=0.001), minimum post-operative lumen area (HR=0.583, 95% CI 0.378-0.879,P=0.010), post-dilatation (HR=0.066, 95% CI 0.013-0.332,P=0.001) and EF>40% (HR=0.019, 95% CI 0.002-0.158, P=0.000). Conclusion: Direct RA had the lower MACCE occurrence rate at 1 year period, this might be related to less operative complication and the optimal lumen gain.

·AIM: To retrospectively analyze the prevalence of retinopathy of prematurity(ROP) in 3471 neonates in Suzhou Municipal Hospital. ·METHODS: A total of 3471 children (1947 males, 1524 females) were screened for ROP in Suzhou Municipal Hospital from January 2010 to September 2016 using binocular ophthalmoscope or ( and) RetCamⅡ. First examination was performed from 4-6wk after birth. The ocular findings were recorded according to the International Classification of ROP and The Early Treatment for ROP. Only the more aggressive eye of bilateral asymmetrical cases was counted for statistical purpose. Children with ROP in both binocular or single eye were counted in 1 case, and the cases required surgeries were defined as severe cases. The prevalence of ROP and severe ROP in recent 6a were analyzed retrospectively. ·RESULTS: The overall relevance ratio of ROP and severe ROP was 17.03% and 1.15%. The relevance ratio of ROP and severe ROP of the males were 16.38% and 1.08%,and of the females were 17.85% and 1.25%, the results were not statistically different (x2= 1. 296, P =0.255). The relevance ratio of ROP and severe ROP of the single birth infants were 17.61% and 1.13%, and of the multiple birth infants were 15.13% and 1.23%,the results were not statistically different (x2=2.706, P=0.100). The children were divided into 5 groups according to the birth weight. The relevance ratio of ROP with birth weight<1000g,1000-1499g,1500-1999g,2000-2499g and ≥2500g were 75. 00%, 36. 17%, 10. 75%, 6. 86% and 3. 77%respectively with significant differences (There were significant differences between the three groups which the birth weight <2000g, P<0.005). The relevance ratio of severe ROP were 36.54%, 1.68%, 0.31%, 0.19% and 0 respectively in these birth weight groups (There were significant differences between the three groups which the birth weight <2000g,P<0.005). The children were divided into 4 groups according to gestational weeks, the relevance ratio of severe ROP of gestational age<28wk,28-31wk, 32-36wk and ≥37wk were 69. 12%, 29. 91%, 8.28% and 3.33% respectively with significant differences (There were significant differences between the three groups which the gestational age <37wk, P<0.005). The relevance ratio of severe ROP were 25%, 1.52%, 0.24% and 0 in these gestational age groups respectively (There were significant differences between the three groups which the gestational age <37wk,P<0.005). · CONCLUSION: The detection rate of ROP in 3471 premature infants was 17. 03%, the severe ROP was 1.15%. There was no evidence that sex and birth were related to ROP, but lower birth weight and smaller gestational age increased the detection rate of ROP.