Objective To prepare Exosomes secreted by mouse hepatoma cell (H_(22)) and heat stressed Exosomes (HS-Exo) derived from heat stress-treated mouse hepatoma cell (H_(22)), in order to study the possible anti-tumor immune mechanism. Methods Exosomes and HS-Exo were purified by serial ultracentrifugation and sucrose density gradiant centrifugation, and were observed and identified by electron microscope. The components and production of the protein and the effects of the host immune response against hepatocellular carcinoma of HS-Exo were observed by using Exosomes as the control. Their immunological factors were detected by Western blot. Lymphocyte proliferation and specific cytotoxic activity of mouse splenic cells were determined by MTT. CD4~+ and CD8~+ lymphocytes infiltration in mouse tumor tissues immunized by both were analysed by immunohistochemical staining. Results HS-Exo was similar in morphology to the Exosomes, the important immune-ralated protein expressed in HS-Exo was increased (P<0.05). HS-Exo immunized mouse group showed more effective inhibition of tumor growth, better-induced lymphocyte proliferation,more significantly enhanced the cytotoxic activity of spleen lymphocytes, as well as a more prominent role in tumor therapy than Exosomes immunized mouse control group (P<0.05). Conclusions Heat stress treatment method for the preparation of HS-Exo was feasible. HS-Exo had a stronger role in the immuneactivity and tumor treatment than control Exosomes.

Objective To evaluate the survival benefit of amphotericin B (AmB) plus flucytosine or fluconazole for treatment of patients with acquired immunodeficiency syndrome (AIDS)-associated cryptococcal meningitis.Methods The following database were searched from the beginning to October 2013,including Cochrane library,PubMed,OVID,Embase,Wanfang Date,CNKI and Chinese Biomedical Database,and the references of eligible studies were manually screened.Reference lists of relevant articles were screened according to selection and extraction criteria.Meta-analysis was performed using RevMan 5.2.Results Four prospective controlled studies with a total of 399 patients with cryptococcal meningitis were identified,including 386 patients with AIDS-associated cryptococcal meningitis and 13 human immunodeficiency virus (HIV)-negative patients.Two hundred and twentyseven patients were treated with AmB and flucytosine combination therapy,including 217 patients with AIDS-associated cryptococcal meningitis and 10 HIV-negative patients.One hundred and seventy-two patients were treated with AmB and fluconazole combination therapy,including 169 patients with AIDS-associated cryptococcal meningitis and 3 HIV-negative patients.The Meta-analysis revealed that the mortality rate in AmB plus flucytosine combination therapy group was 6.6％ (95％ CI:18.5％-31.6 ％) at two weeks point,which was significantly lower than that in AmB plus fluconazole combination group (19.7％,95％CI:-23.6％-62.9％; OR=0.51,95％CI:0.27-0.93,P＜0.05).But at 10 weeks point,the mortality rate in flucytosine combination group was 12.9％ (95％oo CI:-22.2％-48.0％),which was lower than that in fluconazole combination group (31.4％,95％ CI:-23.1％-85.9 ％).However,there was no statistically significant difference between these two groups at 10 weeks point (OR=0.70,95％CI:0.44-1.13,P=0.15).Conclusion Administration of AmB plus flucytosine at early stage can reduce the mortality rate in patients with AIDS-associated cryptococcal meningitis.

Adolescent ; Female ; Humans ; Immunoglobulins, Intravenous ; administration & dosage ; Immunosuppressive Agents ; administration & dosage ; Polyradiculoneuropathy, Chronic Inflammatory Demyelinating ; diagnosis ; therapy

Objective To observe the expression of Na+/I- symporter(NIS) in cultured lactating mammary cells with different levels of iodine and the effect of tumor necrosis factor-α(TNF-α). Methods Original generation of mouse lactating mammary cells cultured in vitro were divided into low iodine group Ⅰ (LI-Ⅰ), low iodine group Ⅱ (LI-Ⅱ), adequate iodine group(AI), high iodine group Ⅰ(HI-Ⅰ), and high iodine group Ⅱ(HI-Ⅱ). Cells were cultured in DEME/F12 culture medium for 24 h with different concentrations of iodine (0,5,50,3000 and 10 000 μg/L, respectively), and TNF-α( 10-2 mg/L) was added to some of cultured cells for 24 h. The expression of NIS mRNA of lactating mammary cells was determined by real-time quantitative PCR and the expression of NIS protein was detected by In-Cell Western. Results In iodine alone group, the expression of NIS mRNA in LI-Ⅰ group [ (64.66 ± 14.99) x 10-4] was higher than that of AI group[ (22.76 ± 7.36) × 10-4, P ＜ 0.01 ]; HI-I group[ (10.18 ±3.53) × 10-4] and HI-Ⅱ group[ (8.59 ± 2.89) × 10-4] were lower than that of AI group(all P ＜ .0.05); With increased iodine concentration, the expression of NIS mRNA decreased. The expression of NIS mRNA in LI-Ⅰ group [(2.72 ± 0.45) × 10-4], LI-Ⅱ group[ (2.69 ± 0.68) × 10-4] and AI group[(1.80 ± 0.67) × 10-4] with iodine plus TNF-o were all lower than that of LI-Ⅰ group, LI-Ⅱ group[ (29.82 ± 4.47 ) × 10-4], and AI group without TNF-α (all P ＜ 0.01). In iodine plus TNF-α, the expression of NIS mRNA in HI-Ⅰ group[(6.58 ± 2.87) × 10-4] and HI-Ⅱ[(7.04 ± 1.36) × 10-4] group were all higher than that of AI group(all P ＜ 0.05); With increased iodine deficiency or iodine excess, the expression of NIS mRNA increased. With increased iodine concentration, the expression of NIS protein decreased in iodine alone group. The expression of NIS protein in iodine plus TNF-α was all lower than that in iodine alone group. In iodine plus TNF-α, the expression of NIS protein increased in both iodine deficiency and iodine excess conditions. Conclusions Iodine may decrease the expression of NIS mRNA and protein of lactating mammary cells. The expression of NIS mRNA and protein of lactating mammary cells was inhibited by TNF-α under different levels of iodine.

ObjectiveTo study the morphological and functional changes of thyroid in lactating rats and their offspring in iodine deficiency and iodine excess animal models.MethodsOne hundred and twenty Wistar rats(30 males and 90 females) were selected.Based on their body weight,the 90 females were stratified and randomly divided into five groups( 18 in each group):low iodine group 1 and group 2(fed with low iodine feed and deionized water containing iodine of 0,5 μg/L) ; high iodine group 1,group 2 and control group(feed with normal diet and deionized water containing iodine of 3000,10 000,50 μg/L).After fed for 3 month,all female rats were mated with males in a ratio of 3 ∶ 1.After birth for 10 days,8 female rats and their offspring in each group were sacrificed.Changes of thyroid were observed by naked eyes.The thyroid weight was measured and pathological changes of thyroids were observed under light microscope.Results①Absolute and relative weight of lactating rats thyroid in low iodine group 1 and group 2 [ (92.02 ± 24.40 ),(77.11 ± 23.32 )mg,(0.509 ± 0.072),(0.384 ± 0.089) mg/kg] were much higher than that of control group[ (17.41 ± 9.25)mg,(0.102 ± 0.016)mg/kg,all P＜ 0.05].Absolute and relative weight of lactating rats thyroid in high iodine group 1 and group 2[(8.22 ± 0.41 ),(9.42 ±0.43)mg,(0.047 ± 0.006),(0.035 ± 0.005)mg/kg] were lower than that of control group(all P ＜ 0.05).Absolute and relative weight of lactating rats and their offspring thyroid was decreased with increase of iodide intake in the diet.②Thyroid enlargement of lactating rats in low iodine group 1 and group 2 was evident,but that of high iodine group 1 and group 2 was not.③Epithelial cell hyperplasia and smaller follicular cavity were observed in low iodine group 1 and group 2 under light microscope.Epithelial cell deformation and mostly flat were observed in high iodine group 1 and group 2.ConclusionsThyroid morphology is changed with iodide intake in the lactating rats and their offspring,and the changes are consistent between female rats and their newborns.

Objective To construct RNAi combinant adenoviral expressive vectors specific to p65 subunit of NF-?B and to observe their gene silencing effect on p65 subunit.Methods Three pairs of complementary. single-strand DNA oligos targeting three various sites of p65 mRNA were designed and synthesized.Annealling was used to generate double-strand oligos(ds-oligos),and then the ds-oligos were cloned into pENTR~TM/u6 to generate the entry clone named pENTR.Recombination reaction in vitro with the pENTR and pAd/BLOCK-iT~TM- DEST was used to creat the adenovirus plasmid which contains the RNAi cassette.Then,the adenovirus plasmids digested with PacI were transfected into HEK293A cells to product adenovirus,and latter infected the HEK293A cells to amplify the adenoviral stock.Plaque forming assay was used to titer the adenoviral stock.The p65 gene silencing effect induced by the RNAi adenovirus was detected by Western blot and immunocytochemistry assay in ECV304 cells.Results The RNAi adenovirus specific to p65 subunit of NF-?B were produced with titer of 3.0 x 10~9pfu/ml to 2.5?10~10pfu/ml.The expression of p65 protein in ECV304 cells could be down-regulated efficiently by the RNAi adenovirus 48-72 h after infection,which would last for more than 6 days after infection.Conclusion RNAi adenovirus is an important tool inhibiting the expression of target gene efficiently.

AIM To investigate the anti-neuroinflammation effects of 4-hydroxybenzyl aldehyde (4-HBAL) from Gastrodia elata Blume on acute cerebral ischemic injury in rats and its nechanism of action.METHODS The rat model of acute cerebral ischemic injury was induced by injecting arachidonic acid via intracarotid artery.Brain tissue samples were taken from the animals 3 h after the model of acute cerebral ischemic injury.Tumor necrosis factor-alpha (TNF-α) and interleukin-1 beta (IL-1β) were detected in brain tissue to evaluate the effects of 4-HBAL in vivo.Lipopolysaccharid (LPS)-induced activation of BV-2 microglia cells model was used to explore the anti-neuroinflammation mechanism of 4-HBAL.RESULTS The experimental results showed that 4-HBAL had a significant protective effect on acute cerebral ischemic injury.It could significandy decrease the contents of tumor necrosis factor-alpha (TNF-α) and interleukin-1 beta (IL-1β),and obviously inhibit the production of nitric oxide (NO),prostaglandin E2 (PGE2) and TNF-α in LPS-stimulated BV-2 cell,and increase the production of interleukin-10 (IL-10) and transforming growth factors-β (TGF-β) in BV-2 cell.CONCLUSION The mechanism of 4-HBAL may be related to the suppression of the excessive activation of microglia after cerebral ischemia and the promotion of the transformation of microglia into anti-inflammatory phenotype.

Objective To discuss the diagnostic value of MRI for portal vein(PV)and superior mesenteric venous(SMV)thrombosis.Methods Twelve patients with portal vein and superior mesenterie venous thrombosis proved by operation and pathology were examined with T_1WI,T_2WI,T_2-weighted fat suppression imaging,MR angiography(MRA)and Gd-DTPA enhanced dynamic MRI.Results Signals in PV and SMV were detected on T_1 WI and T2WI in 12 cases;3 acute thrombus presented hypo-or isointense on T_1 WI and hyperintense on T_2 WI.ttyperintense on T_1 WI and T_2WI were showed in 8 subacute thrombus; 1 chronic thrombus presented heterogenous intense on T_1W1 and hypointense on T_2W1.No enhancement within PV and SMV was found on Gd-DTPA enhanced images.Bowel dilatation was found in 10 cases,bowel hemorrhage in 6,bowel wall thickening in 12,intestinal pneumatosis in 3,ascites in 12,cavernous transformation of the portal vein in 3,hepatic perfusion disorder in 6.Conclusion MRI is an important and sensitive imaging method for the diagnosis and location of portal vein and superior mesenteric venous thrombosis with intestinal ischemia.

OBJECTIVETo investigate the plasma Big endothelin-1 levels in patients with gastric carcinoma before and after radical gastrectomy, and explore its clinical significance.

METHODSOne hundred and six patients with gastric carcinoma and 20 controls were enrolled. The Big ET-1 plasma levels were examined by enzyme-linked immuno absorbent assay before and on the 1st, 3rd, and 10th day after curative surgery, and then were tested every 3 months in the patients with advanced gastric cancer.

RESULTSAll patients, except those with stage I gastric cancer, had significantly higher mean plasma Big ET-1 levels compared with normal controls (P=0.000). Higher plasma Big ET-1 levels were associated with lymph node metastasis (P=0.020) and serosal infiltration (P=0.035). The plasma Big Endothelin-1 levels were markedly increased on the first post-operative day (1st POD) in all patients,but decreased on the 3rd POD with no significant difference compared to the preoperative levels. On the 10th POD, the patients with stage I and II gastric cancer showed marked reduction in plasma Big ET-1 levels (P=0.010 and P=0.000, respectively), whereas no significant difference was observed in stage III and IV patients. During the follow-up, the plasma Big ET-1 levels just before recurrence in stage II patients were significantly higher compared with the levels on the 10th POD (P=0.011).

CONCLUSIONSPlasma Big ET-1 might be a reliable marker to determine the severity of gastric carcinoma. Monitoring plasma Big ET-1 levels after curative resection in stage II gastric cancer patients is valuable to predict recurrence.

Adult ; Aged ; Case-Control Studies ; Endothelin-1 ; blood ; Female ; Follow-Up Studies ; Gastrectomy ; Humans ; Male ; Middle Aged ; Neoplasm Recurrence, Local ; pathology ; Neoplasm Staging ; Stomach Neoplasms ; blood ; pathology ; surgery

ObjectiveTo observe the influence of iodine on mRNA expression of iodide transporter (NIS),insulin-like growth factor Ⅰ (IGF- Ⅰ ) and transforming growth factor beta(TGF-β) in thyroid and mammary glands of lactating rats and to explore the role of NIS,IGF- Ⅰ and TGF-β mRNA in iodine uptake in thyroid and mammary glands of lactating rats.MethodsOne hundred and one Wistar rats(80 female and 21 male),weighting 8 - 100 g were selected.These female rats were randomly divided into five groups according to their body weight:control group(NI,normal feed,drank deionized water containing iodine 50 μg/L) ; low iodine group 1 and 2(LI-1,LI-2,low iodine feed,drank deionized water containing iodine 0 and 5 μg/L,respectively); high iodine group 1 and 2(HI-1,HI-2,normal feed,drank deionized water containing iodine 3000 and 10 000 μg/L,respectively),16 rats in each group.After feeding for 3 months,the female and male rates were mated 3:1.The female rats in each group were sacrificed at the fifth and tenth day after postpartum.Thyroid and mammary glands were taken.The mRNA levels of NIS,IGF- Ⅰ and TGF-β in thyroid and mammary glands of lactating rats were determined by real time quantitative PCR.ResultsThe fifth days after postartum,NIS,IGF- Ⅰ and TGF-β mRNA expression levels of thyroid and lactating mammary glands were different between groups,and the differences were statistically significant ( NIS:F =631.46,64.91,all P ＜ 0.01 ; IGF- Ⅰ:F =11.45,6.56,all P ＜ 0.01 ; TGF-β:F =291.83,304.53,all P ＜ 0.01).Compared with control group [NIS:0.0066 ± 0.0023, (0.1481 ± 0.0711 ) × 10-2; IGF- Ⅰ:0.0419 ± 0.0062,0.0542 ± 0.0044; TGF-β:0.1416 ± 0.0277,0.1670 ± 0.0499],regardless of thyroid or mammary gland,the NIS,IGF- Ⅰ and TGF-β mRNA expression of LI-1 [NIS:0.0447 ± 0.0110,(0.3030 ± 0.1831) × 10-2;IGF- Ⅰ:0.0662 ± 0.0078,0.0902 ± 0.008; IGF- Ⅰ:0.5514 ± 0.0508,0.6942 ± 0.0367],LI-2[NIS:0.0317 ±0.0081,(0.3017 ± 0.1601) × 10-2; IGF-I:0.0645 ± 0.0054,0.0894 ± 0.0093; TGF-β:0.5292 ± 0.0332,0.6704 ± 0.0277 ] was significantly increased (all P ＜ 0.01 ); the NIS mRNA expression of HI-1 [0.0043 ± 0.0011,(0.1233 ± 0.0954) × 10-2],HI-2[0.0037 ± 0.0017,(0.1058 ± 0.0854) × 10-2] was decreased(all P ＜ 0.05),while the expression of IGF-Ⅰ mRNA [0.0521 ± 0.0910,0.0715 ± 0.0026; 0.0516 ± 0.0078,0.0697 ± 0.0038] and TGF-β mRNA [0.2087 ± 0.0425,0.2361 ± 0.0425; 0.1971 ± 0.0237,0.2257 ± 0.0752 ] was increased (all P ＜ 0.05 ).The tenth days after postpartum,the mRNA expression levels of NIS,IGF- Ⅰ and TGF-β of thyroid and lactating mammary gland in rats were different between groups,and the differences were statistically significant (NIS:F =103.55,116.32,all P ＜ 0.01; IGF-Ⅰ:F =67.67,11.98,all P ＜ 0.01; TGF-β:F =74.30,381.30,all P ＜0.01 ).Compared with the control group[NIS:0.0069 ± 0.0011,(0.1337 ± 0.0599) × 10-2; IGF-Ⅰ:0.0390 ±0.0071,0.0534 ± 0.0056; TGF-β:0.1351 ± 0.0336,0.1534 ± 0.0320],the mRNA expression levels of NIS,IGF- Ⅰ and TGF-β of LI-1 [ NIS:0.0432 ± 0.0165,(0.2962 ± 0.0985 ) × 10-2; IGF- Ⅰ:0.0643 ± 0.0088,0.0873 ± 0.0055 ; TGF-β:0.5042 ± 0.0912,0.6408 ± 0.0420],LI-2[NIS:0.0287 ± 0.0111,(0.2873 ± 0.0862) × 10-2; IGF- Ⅰ:0.0621 ± 0.0094,0.0862 ± 0.0038; TGF-β:0.4893 ± 0.0504,0.6372 ± 0.0389] were significantly increased(all P ＜ 0.01 ); the NIS mRNA levels of HI-1 [ 0.0042 ± 0.0029,(0.1006 ± 0.0909) × 10-2],HI-2[0.0035 ± 0.0020,(0.0890 ± 0.0119) × 10-2] were decreased(all P＜ 0.05),while the expression of IGF-Ⅰ mRNA[0.0516 ± 0.0078,0.0668 ± 0.0071; 0.0508 ± 0.0089,0.0621 ± 0.0064] and TGF-β mRNA[0.2007 ± 0.0546,0.2175 ± 0.0370;0.1959 ± 0.0393,0.2097 ± 0.0425] were increased(all P ＜ 0.05 ).In thyroid and mammary glands,the comparisons of NIS,IGF,TGF-β mRNA expression of the fifth and tenth day after postartum,between each group were not statistically significant(all P ＜ 0.05).ConclusionsThere are regulatory mechanisms of thyroid and mammary glands of lactating rats in response to low or high iodine conditions.In low iodine,the expressions of NIS,IGF- Ⅰ and TGF-β mRNA in thyroid and mammary glands increase and iodide uptake ability is enhanced to meet the body needs.In high iodine,the expression of NIS mRNA decreases in thyroid and mammary glands.Due to the reduced ability of iodine uptake,iodine intake is reduced,thereby reducing the hazards of high iodine in filial rats.