OBJECTIVETo observe mainfestations of syndrome and biochemical indices of hypertensive model rats with excessive accumulation of phlegm-dampness syndrome (EAPDS), and to explore its possible pathological mechanism.

METHODSEAPDS rat model was prepared in 50 Wistar rats by feeding with high fat forage. Meanwhile, a normal control group consisting of 10 Wistar rats was set up by feeding with normal forage. After 25-week continuous feeding, 22 rats with body weight (BW) and blood pressure (BP) exceeding 25% those of the control group were selected as a model group. BW, BP, blood lipids, and related serological indicators were detected in all rats. Morphological changes of target organs were observed. mRNA expression levels of leptin receptor (LepR), Janus kinase2 (Jak2), signal transducer and activator of transcription 3 (Stat3), suppressor of cytokine signaling-3 (Socs3), angiotensin II receptor type 1 (AT1), angiotensin II receptor type 2 (AT2), phosphatidylinositol 3 kinase (P13K), serine threonine kinase (Akt), nuclear factor of kappa B (NF-κBp65), inhibitor of nuclear factor kappa-B kinase α (IKKα), NF-kappa-B inhibitor β (lKKβ), NF-kappa-B inhibitor α (IKBα), and AMP-activated protein kinase (AMPK) were detected by quantitative real-time PCR (qPCR). Expression levels of AT1 and LepR in aorta were detected by immunohistochemical assay and Western blot respectively.

RESULTSCompared with the control group, BW, BP, and blood lipids increased; serum levels of leptin (Lep) , Ang II, Hcy, ET-1, TNF-α, IL-6, and p2-MG increased, but NO decreased in the model group (P < 0.05, P < 0.01). Aortal endothelial injury and smooth muscle cell proliferation occurred in the model group, accompanied with heart and renal injury. Compared with the control group, mRNA expression levels of LepR, Jak2, Stat3, Socs3, AT1 , PI3K, Akt, NF-κB p65, IKKβ, IKBα, and AMPK in aorta were up-regulated significantly (P < 0.05), while the expression of IKKa decreased (P < 0.05). Immunohistochem- ical staining showed, brownish yellow deposit of AT1 and LepR was obviously increased, with more extensively positive distribution. Western blot results showed, as compared with the control group, protein expression levels of AT1 and LepR obviously increased in the model group (P < 0.05).

CONCLUSIONSModel rats exhibited typical syndromes of EAPDS. They put up weight with fat abdomen, gloomy hair, poor appetite, hypersomnia, lowered activities , reduced food intake, loose stool, dark red tongue, white tongue with white, thick, greasy fur. Lep could be taken as one of objective indicators for evaluating hypertension rat model with EAPDS.

Animals ; Aorta ; Cell Proliferation ; Disease Models, Animal ; Hypertension ; physiopathology ; I-kappa B Proteins ; Interleukin-6 ; Leptin ; blood ; NF-KappaB Inhibitor alpha ; NF-kappa B ; Phosphatidylinositol 3-Kinases ; Rats ; Rats, Wistar ; Suppressor of Cytokine Signaling Proteins ; Transcription Factor RelA ; Tumor Necrosis Factor-alpha

Objective Investigated the treatment effect of ISCOM leukemia vaccine combination with 1-methyl tryptophan on tumor burden mice .Methods Saponin was added lipase protein (1 mg/mL) 7 ℃ for 12 h ,adding 80 μL lipid mixed solution and 5 mL saponin solution (1 mg/mL ) to prepare ISCOM leukemia vaccine .C57BL /6 mice were randomly divided into model group , ISCOM leukemia vaccine group ,1-methyl tryptophan group and combination group ,Mice were injected FBL-3 cell to built leukemia tumor-burdened mice model .After treatment for 4 weeks ,tumors weight ,NK and Mφ and CTL cell killing activity ,serum levels of IL-10 ,IL-12 were detected .Results Tumor weight in combination group was less than 1-methyl tryptophan and ISCOM leukemia group [(0 .64 ± 0 .26)g vs .(2 .49 ± 0 .91)g ,P< 0 .01 ,(0 .64 ± 0 .26)g vs .(1 .28 ± 0 .73)g ,P< 0 .05] ;NK cell killing activity in com-bination group was higher than 1-methyl tryptophan group[(38 .41 ± 8 .27)% vs .(67 .22 ± 12 .74)% ,all P< 0 .01)] ;M φ activity in combination group was significantly higher than 1-methyl tryptophan group[(55 .69 ± 13 .69)% vs .(69 .47 ± 14 .79)% ,P< 0 .01] ;CTL activity in combination group was significantly higher than 1-methyl tryptophan group and ISCOM leukemia group[(43 .77 ± 8 .89)% vs .(69 .68 ± 11 .44)% ,P< 0 .01 ,(58 .87 ± 9 .45)% vs .(69 .68 ± 11 .44)% ,P < 0 .05] ;IL-10 in combination group were significantly lower than 1 - methyl tryptophan group and ISCOM leukemia group [(76 .2 ± 6 .82)pg /L vs .(98 .3 ± 13 .4)pg/L ,P<0 .01 ,(76 .2 ± 6 .82)pg/L vs .(202 .3 ± 44 .5)pg/L ,P < 0 .01] ;IL-12 in combination group were significantly higher than 1-methyl tryptophan group and ISCOM leukemia group[(381 .2 ± 47 .3)pg/L vs .(332 .1 ± 30 .2)pg/L ,P < 0 .05 ,(381 .2 ± 47 .3)pg /L vs . (291 .2 ± 17 .3)pg/L ,P< 0 .01] .Conclusion Combination with 1-methyl tryptophan and ISCOM leukemia vaccine has a well anti-tumor effect ,its mechanism may be through mediated and the expression of IL-12 and IL-10 .

Objective To compare the effects of biphasic square waveform (BSW) with low or high energy on external defibrillation.Method Adult swine model of closed chest ventricular fibrillation induced by electricity was established.Eighteen swine,weighing (30?3.3) kg were randomly divided into three groups:50-50-50 J group (n=6),30-50-75 J group (n=6),120-150-200 J (n=6).After three minutes of ventricular fibrillation without treatment,the pigs in the three groups were defibrillated accordance to the above sequences. Results 30 J BSW didn't succed to external defibrillate.The first defibrillation successful rate of 50 J and 120 J BSW was 5/6.The total defibrillation successful rate of every group was 100%.All pigs quickly had spontaneous circulation after defibrillation and survived more than 24 hours.ST-T change of low-energy was less than that of high-energy.After resuscitation,myocardial function decreased,but there had not significant differences between groups.Conclusions In the study,30J BSW could not reach successful defibrillation,and 50 J and 120 J BSW had similar defibrillation efficacy.The ideal energy of BSW external defibrillation was 50 J.

The synthesized full-length hGLP-1 gene was cloned into pET-32a(+) to get the recombinant plasmid pET32-GLP-1, which could express a fusion protein containing thioredox, hexahistidine, and rhGLP-1 consecutively. The recombinant plasmid containing hGLP-1 was transformed into E.coli BL21 (DE3) and expressed by IPTG induction. The fusion protein was purified from lysates with Ni?IDA His?Bind affinity chromatography. rhGLP-1 with the purity of 90% was achieved after enterokinase digestion, Ni?IDA His?Bind affinity chromatography again, then was concentrated by ultrafiltration. The purified rhGLP-1 showed a single band on IEF gel with an isoelectric point between pH5.2 and pH5.85. ESI mass spectrometry showed that the molecular weight was 3355.0kDa as expected. rhGLP-1 was digested with trypsin followed by mass analysis and the peptide mapping produced two expected fragments with the molecular weights of 2097.7kDa and 1005.5kDa, respectively. The purified rhGLP-1 also showed obvious biological activity for both lowering plasma glucose and stimulating insulin secretion in mice.

Objective To study the osteogenic effects of induced endothelial cell(EC)on bone marrow stromal cells(BMSCs)of rabbits in co-culture condition.Methods BMSCs were obtained from rabbits by density gradient centrifugation.The adhesive ceils were preserved to passage in culture.The cultured cells were divided into four groups:group A(BMSCs),group B(BMSCs osteogenic induction),group C(EC induction)and group D (co-culture of induced BMSCs and EC).The cell morphology,immunofluorescence,cell proliferation,alkaline pbosphatase(ALP)activity,osteocalcin synthesis were observed to determine the effects of induced autologous EC on the osteogenic potential and cellular compatibility of BMSCs.Results The immunochemical staining showed that the BMSCs were induced into EC in group C.The cellular compatibility of BMSCs and EC was good.The ALP activity and osteocalcin content were obviously higher in group D than in any other groups(P＜0.05).The cell proliferation difference was not obvious between groups(P＞0.05).Conclusions The cellular compatibility of induced osteoblasts and induced EC is perfect.The ECs can significantly increase the viability and ALP activity of induced osteoblasts.

Emerging data indicated that HCV subverts the antiviral activity of interferon (IF); however,whether HCV core protein contributes to the process remains controversial. In the present study, we examined the effect of HCV core protein on interferon-induced antiviral gene expression and whether the effect is involved in the activation and negative regulation of the Jak/STAT signaling pathway. Our results showed that, following treatment with IFN-α, the transcription of PKR, MxA and 2'-5'OAS were down-regulated in HepG2 cells expressing the core protein. In the presence of HCV core protein,ISRE-dependent luciferase activity also decreased. Further study indicated that the core protein could inhibit the tyrosine phosphorylation of STAT1, whereas the level of STAT1 expression was unchanged.Accordingly, SOCS3, the negative regulator of the Jak/STAT pathway, was induced by HCV core protein. These results suggests that HCV core protein may interfere with the expression of some interferon-induced antiviral genes by inhibiting STAT1 phosphorylation and induction of SOCS3.

Cephalosporin is still the first-line drug chosen for the treatment of gonorrhea, although it is likely to develop resistance to Neisseria gonorrhoeae. Recently, the resistance or low sensitivity of cephalosporin to Neisseria gonorrhoeae have been reported at home and abroad. The studies of resistance mechanism to Neisseria gonorrhoeae are particularly important. The resistance mechanism is mainly lied in three aspects. The first is the changes in antibiotic target sites, for example, the changes in penicillin-binding proteins would affect the binding of antibiotics to Neisseria gonorrhoeae; the second is the changes in bacterial cell membrane porin which effects drugs influxing; the third is the changes in efflux system which enhances the bacterial efflux capacity. To study and explore the mechanism of Neisseria gonorrhoeae resistance is of great importance for the prevention and treatment of gonorrhea.

OBJECTIVETo investigate the impact of omeprazole on platelet response to clopidogrel and the effect of polymorphisms of CYP2C19 on the antiplatelet effect of clopidogrel.

METHODSPlatelet aggregation (PA) was assessed before 300 mg aspirin plus 300 mg loading dose of clopidogrel and after 300 mg aspirin plus 75 mg maintenance dose of clopidogrel 7 days later in 414 patients with acute coronary syndrome who have undergone percutaneous coronary intervention (PCI). Thereafter, gastric mucosal protective drugs were given (omeprazolem 20 mg, n=224 or cimetidine 800 mg, n=190). Fourteen days later, PA was measured again. Genotypes of CYP2C19*2 were analyzed with polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP).

RESULTSAfter taken aspirin and clopidogrel, PA has decreased significantly in both groups. Compared with cimetidine, omeprazole had no significant impact on PA on 7 and 21 days post PCI. Compared with homozygotes or heterozygotes for the wild-type CYP2C19*2, patients with CYP2C19*2 AA genotype had significantly higher PA on 7 and 21 days post PCI (P<0.05).

CONCLUSIONNo attenuating effect on platelet response to clopidogrel has been observed for Omeprazole. The variant of CYP2C19*2 AA genotype is significantly associated with attenuated response to clopidogrel.

Adult ; Aged ; Aryl Hydrocarbon Hydroxylases ; genetics ; metabolism ; Cytochrome P-450 CYP2C19 ; Drug Interactions ; Female ; Humans ; Male ; Middle Aged ; Omeprazole ; pharmacology ; Platelet Aggregation Inhibitors ; pharmacology ; Proton Pump Inhibitors ; pharmacology ; Ticlopidine ; analogs & derivatives ; pharmacology

OBJECTIVETo determine the effects of KH2PO4 on the odonto- and osteogenic differentiation potential of human stem cells from apical papillae (SCAP) in vitro.

METHODSSCAP were isolated and cultured respectively in alpha minimum essential medium (α-MEM) or α-MEM containing 1.8 mmol/L KH2PO4. Alkaline phosphatase (ALP) activity, alizarin red staining, real-time reverse transcription polymerase chain reaction (RT-PCR) and Western blotting were used to examine the odonto and osteogenic potential of SCAP in the two media.

RESULTSSCAP cultured in α-MEM containing 1.8 mmol/L KH2PO4 exhibited a higher ALP activity [(0.370 ± 0.013) Sigma unit×min(-1)×mg(-1)] at day 3 than control group [(0.285 ± 0.008) Sigma unit×min(-1)×mg(-1)] and KH2PO4-treated SCAP formed more calcified nodules at day 5 [(0.539 ± 0.007) µg/g] and day 7 [(1.617 ± 0.042) µg/g] than those in normal medium [(0.138 ± 0.037) µg/g, P < 0.01]. The expression of odonto- and osteogenic markers were significantly up-regulated after the stimulation of KH2PO4 at day 3 and 7 respectively, as compared with control group.

CONCLUSIONS1.8 mmol/L KH2PO4 can promote the odonto and osteogenic differentiation potential of human SCAP.

Cell Differentiation ; drug effects ; Cells, Cultured ; Dental Pulp ; cytology ; Extracellular Matrix Proteins ; metabolism ; Humans ; Osteoblasts ; cytology ; Osteocalcin ; metabolism ; Phosphates ; pharmacology ; Phosphoproteins ; metabolism ; Potassium Compounds ; pharmacology ; Sialoglycoproteins ; metabolism ; Stem Cells ; cytology ; drug effects ; metabolism

OBJECTIVETo investigate the influence of CD4+ CD25+ regulatory T cells(Treg cells) on mouse gastric cancer.

METHODSTreg cell in mouse spleen bearing gastric tumor was tested in different time points. Magic cell sorting(MACS) method was used to purify mouse Treg cells and the Treg cells were injected into mouse bearing gastric tumor with different dosage. After 3 weeks, the tumor size and tumor cell apoptosis rate were measured.

RESULTSTreg existed in normal mouse spleen with a rate of (3.86+/-0.07)%. In tumor model this percentage increased gradually and was (4.12+/-0.13)% after 3 weeks, which was significantly higher than that in control. When Treg cell applied in mouse reached 2.0 x 10(5), the tumor size enlarged significantly(P=0.013) and tumor cell apoptosis rate decreased significantly (P=0.012).

CONCLUSIONSTreg cell is associated with gastric cancer progress in mouse tumor model. Treg cell can promote gastric cancer growth and decrease tumor apoptosis. The anti- Treg GITR can improve anti- tumor effects.

Animals ; Apoptosis ; Female ; Flow Cytometry ; Male ; Mice ; Mice, Inbred Strains ; Spleen ; cytology ; Stomach Neoplasms ; immunology ; pathology ; T-Lymphocytes, Regulatory ; immunology