Objective To investigate the effect of specific small interfering RNA (siRNA) targeting receptor of advanced glycation end products (RAGE) on the production of fibrosis markers (laminin,hyaluronic acid (HA) and N-terminal procollagen Ⅲ propeptide (PⅢ NP) in hepatic fibrosis (HF).Methods The expression vectors of specific siRNA targeting RAGE were constructed.Primary rat hepatic stellar cells (HSC) were isolated and cultured.The primary rat HSC were transfected with the recombinant vector.The blank control group and unspecific siRNA vector pAKD-NC-transfected group were as controls.The expressions of RAGE,laminin,HA and PⅢ NP at mRNA and protein levels were detected by real-time polymerase chain reaction and Western blot,respectively.Least-significant difference (LSD) and Student-Newman-Keuls (SNK) were performed to analyze standard normal distribution or homogeneous variance.Non-normal distribution and heterogeneity of variance data were analyzed by non-parametric Wilcoxon test.Results The expressions of RAGE at mRNA and protein levels in pAKD-GR126-transfected primary HSC were (42.32 ± 6.16)％,(43.24±7.50)％,(51.06±13.79)％ and (47.94±5.36)％ in blank control group and pAKDNC group (F=7.791 and 36.513,all P＜0.05).The expressions of laminin at mRNA and protein levels were (41.07±3.13)％,(40.59±5.87)％,(53.89±2.25)％ and (52.46±4.68)％ in blank control group andpAKD-NC group (F=225.111 and 88.039,all P＜0.05).The expressions of HA at mRNA and protein levels were (45.69 ± 0.87) ％,(46.08 ± 2.36) ％,(54.20 ± 0.56) ％ and (52.30±3.42)％ in blank control group and pAKD-NC group (F=178.317 and 180.646,all P＜ 0.05).The expressions of PⅢ NP mRNA at mRNA and protein levels were (56.10±4.18)％,(55.15±2.39)％,(54.40±2.79)％ and (53.58±6.18)％ in blank control group and pAKD-NC group (F=141.633 and 49.670,all P＜0.05).Conclusion RAGE specific siRNA could inhibit the expression of RAGE in primary rat HSC and could significantly lower the expression of fibrosis markers laminin,HA and PⅢ NP at mRNA and protein level.

This article reviewed the researches proceeding on the contamination and detection of the foodborne pathogen noroviruses (NoVs) in fresh produce, which involved the NoVs contaminations in fresh produce, the special attachment of NoVs in fresh produce, the NoVs outbreaks associated with fresh produce and the NoVs detection in fresh produce. There had been an increase in reported infectious disease risks associated with the consumptions of fresh produce for recent 30 years. Because the NoVs, as a primary cause of viral gastroenteritis thoughout the world, were highly contagious, had a low infectious dose, and were persistent in the environment. And also the methods for NoVs detection in food had significantly developed over the last 15 years. Currently NoVs were the most common pathogen accounting for 40% of outbreaks associated with fresh produce (i. e., fruits and vegetables). Data from outbreaks investigations verified fresh produce as the high risk food products for NoVs. The fresh produce were typically eaten raw with no thermal processing, can be contaminated at any step during production and processing from faecally polluted water and fertilizers, the poor hygiene practices by food handlers and the cross-contamination. The attachment of NoVs to the fresh produce was due to the physio-chemical factors of virus protein coat, the special attachment to different fresh produce, and the possibility for internalization of NoVs. It might provide answers to why those high risk foods were more frequently implicated (i. e., lettuce and raspberries). According to the data of foodborne NoVs outbreaks which were associated with fresh produce from EU countries and the USA, the outbreaks in EU countries were mainly associated with NoVs contaminated raspberries and lettuce, while in USA which were associated with NoVs contaminated lettuce. Unfortunately, there were no NoVs detection methods for fresh produce or the data of foodborne NoVs outbreaks which were associated with fresh produce in China. That made it difficult to analyze the NoVs contamination situation in China. The heterogeneous distributions of presumably low levels of virus, which presented in contaminated fresh produce, also made it difficult to detect NoVs. To solve this problem, different sampling methods, viral elution methods and RT-qPCR methods were chosen. For example, according to the isoelectric point of NoVs particles, high pH and high ionic strength solution could be used as means for releasing NoVs. For the elution from acidic fruit, the buffer capacity and the virus recovery could be increased by the addition of tris-HCl. When analyzing pectin containing raspberries or strawberries, the viral elution usually incubated with pectinase at neutral pH to avoid from foaming jelly. In this paper, the latest ISO standard for NoV detection in food and the new approaches for NoV detection were also reviewed to provide references for domestic researches. It was necessary to establish and develop domestic methods for NoV detection in fresh produce, especially the different NoV conventional molecular detection methods with corresponding NoV extraction methods, which targeted to the different adsorption characteristics of different fruits and vegetables, in order to strengthen the national food safety monitoring.
Food Analysis ; methods ; Food Contamination ; analysis ; Foodborne Diseases ; virology ; Fruit ; virology ; Gastroenteritis ; virology ; Humans ; Norovirus ; classification ; genetics ; isolation & purification ; physiology ; Vegetables ; virology

Objective To observe the effect of everolimus on proliferation,cycle and apoptosis of endometrial cancer cells and discuss the mechanism.Methods Proliferation of RL95-2 interfered with everolimus in different concentration and time was observed by MTT assay.Cell cycle and apoptosis affected by everolimus was detected by flow cytometry.PI3K-Akt-mTOR signaling pathway was detected by Western blot.Results (1) MTT assay showed that,cell viability in everolimus group was time-dependent decreased,compared with control group,there were significant differences in 48 h,72 h and 96 h ((64.36±4.78)％,(48.18±5.93) ％,(42.72±6.91) ％;F inner grouP =3.278,P＜ 0.05;F between groups =12.327,P＜ 0.01;F across groups=7.729,P＜0.05).Cell viability in everolimus group was dose-dependent decreased(F=3.264,P＜0.05).Compared with control group,RL95-2 cells in G1 phase and G2 phase was (69.28±2.61)％ and (4.75±0.84) ％,decreased in everolimus group,and in S phase (27.31±0.69) ％ was increased (t=5.743,P＜0.05;t =4.528,P＜0.05;t=6.209,P＜0.05).The apoptosis in everolimus group was 29.78％,higher than in control group (47.29％),the difference was significant (t =19.381,P＜0.01).(2) Western blot showed that both mTOR and p-Akt in everolimus group were dose-dependent decreased (F=3.589,P＜0.05;F =5.292,P＜0.05),Akt was dose-dependent increased (F =4.294,P＜0.05).Conclusion Everolimus decrease the cell viability and promote apoptosis via target inhibiting mTOR expression.

Objective To observe the expression of ZNF139 and MMP-7 in endometrial adenocarcinoma tissues and discuss the correlation with clinical pathological significances.Methods In 71 patients diagnosed as endometrial adenocarcinoma,71 cancer tissues(adenocarcinoma group) and 71 adjacent normal tiuues(adjacent cancer group) were collected in operations.Immunohistochemical methods was used to detect the expression of ZNF139 and MMP-7 in the two groups,the relationship between the two genes and communication with clinicopathological indexes were discussed.Western blot and qRT-PCR were used to detected the expression of ZNF139 and MMP-7 at protein and mRNA level.Results The positive rates of ZNF139 was 66.20% in adenocarcinoma group,higher than in adjacent cancer group(21.42%,P=0.016);the positive rates of MMP-7 was 74.65% in adenocarcinoma group,higher than7 than in adjacent cancer group(26.76%,P=0.012).The expression of ZNF139 and MMP-7 was correlated with lymph node metastasis,muscle invasion and FIGO stage(P=0.028,0.031;P=0.004,0.016;P=0.008,0.011).Spearman rank correlation analysis showed there was positive correlation between ZNF139 and MMP-7 in adenocarcinoma tissues(r=0.716,P=0.039).Western blot showed that ZNF139 and MMP-7 expression in adenocarcinoma group were higher than in adjacent cancer group(t=6.92,7.34;P<0.01);qRT-PCR showed that ZNF139 mRNA and MMP-7 mRNA expression in adenocarcinoma group were higher than in adjacent cancer group(t=4.27,4.06;P<0.05).Conclusion ZNF139 and MMP-7 are correlated with malignant pathological characteristics in endometrial adenocarcinoma,they may modulate the malignant behavior together.

Objective To investigate the association between matrix metalloproteinase 9(MMP 9) and type 2 diabetic macroangiopathy and explain the mechanism of Simvastatin protective effects on blood vessel.Methods The serum MMP 9 was determined by ELISA in 40 healthy controls and 80 type 2 diabetes(including 40 type 2 diabetic macroangiopathy and 40 type 2 diabetes without complications) before and after Simvastatin treatment.Results The serum MMP 9 in type 2 diabetic macroangiopathy was higher than that in type 2 diabetes without complications and healthy controls.Serum MMP 9 was correlated positively with TC,LDL C,Ox LDL and blood pressure and correlated negatively with HDL C. After Simvastatin treatment,the levels of serum MMP 9 were reduced and serum TC,LDL C and Ox LDL were also decreased.Conclusion MMP 9 may play an important role in the development and progression of type 2 diabetic macroangiopathy.The protective effects of Simvastatin on blood vessel partly attribute to its lowering of serum MMP 9.

Objective To compare the value of 99m Tc-DTPA dynamic renal imaging and intravenous pyelography(IVP) in assessing renal function. Methods The results of 99m Tc-DTPA dynamic renal imaging and IVP in 117 patients with renal diseases confirmed by clinical examinations, abdominal ultrasonography, CT and blood biochemical markers were analysed retrospectively. Results Out of 234 kidneys,101 were clearly visualized by IVP, which were indicated normal renal uptake by 99m Tc-DTPA dynamic renal imaging, and the GFR value was (60.13?16.85)ml/min. 48 kidneys were poorly visualized by IVP, their uptake detected by dynamic renal imaging was between grade 1 to 3, and the GFR value was (23.06?10.61)ml/min. 85 kidneys were not visualized by IVP, 71.8% of which(61/85kidneys) were indicated uptake of radionuclide by dynamic renal imaging with the (20.39?12.54)ml/min GFR value, and uptake of 8 kidneys among the 61 kidneys was normal. The functional kidneys detected by dynamic renal imaging were more than those detected by IVP(P

Objective:To study the changes of the NKT like cells after HIV infected. Methods:We collected peripheral blood from 47 untreated HIV infected individuals and 31 healthy controls,and analyzed the expression of Annexin-V,Ki-67,HLA-DR and other surface molecules in NKT like cells by flow cytometry. Results:The NKT like cell percentage of untreated HIV infected group was (3. 03±1. 61)%,the NKT like cell percentage of normal control group was (8. 30±7. 42)%,the percentage of NKT like cells in HIV infected individuals was significantly lower than the healthy controls ( P<0. 05 );the NKT like cell HLA-DR expression of untreated HIV infected group and normal control group were (5. 40±4. 10)% and (0. 89±0. 83)%,the NKT like cell Annexin-V expression of untreated HIV infected group and normal control group were (30. 21±13. 15)% and (5. 40±8. 05)% ,and the activation and apoptosis of NKT like cells was significantly higher after HIV infection compared with health individuals (P<0. 001,P<0. 01),the degree of activation was negatively correlated with CD4 count (r=-0. 885 7,P<0. 05);and the NKT like cell Ki-67 expression of untreated HIV infected group and normal control group were (11. 15±4. 76)% and (27. 63±18. 31)%,the proliferation ability was significantly lower after HIV infection compared with healthy controls(P<0. 05). Conclusion:HIV infection can significantly reduce the number of NKT like cells and its ability to proliferate,and increase its ability to activation and apoptosis.

Objective To test thyroid-stimulating hormone (TSH) suppress GLUT4 expression and translocation by stimulating TNF-α secretion in 3T3-L1 adipocytes via a cAMP-PKA pathway.Methods 3T3-L1 preadipocytes were induced to differentiate into adipocytes.The adipocytes were treated with bovine TSH,Forskolin,H89 and Rapamycin,respectively.The concentration of TNF-α in the cell culture medium was measured by ELISA.The level of GLUT4 mRNA in adipocytes was assessed by real time polymerase chain reaction.Protein levels of GLUT4 in total cell lysates and plasma membrane lysates were quantified by Western blotting.Results Incubating 3T3-L1 adipocytes with TSH markedly increased the concentration of TNF-α in medium in a time-and dosedependent manner (P ＜ 0.05); meanwhile,the levels of GLUT4 mRNA and total and plasma membrane GLUT4 protein were decreased in a dose-dependent manner (P＜0.05 or ＜0.01).H89 and rapamycin could block the above effects respectively (326.7±43.2 vs.341.9±12.0,P＞0.05).However,there was no statistical difference in the TNF-α levels between stimulation with 1 μmmol/L forskolin versus 0.04 μmmol/L bovine TSH (481.9± 28.4 vs.522.7± 36.2,P＞0.05).Conclusions TSH can down-regulate GLUT4 expression and translocation in 3T3-L1 adipocytes by stimulating TNF-α secretion through a cAMP-PKA pathway.

Oxytocin ( OT ) is a cyclic neuropeptide containing nine amino acids residues, in addition to the traditional roles of uterine contraction and lactation, it also plays important roles in the central nervous system and other peripheral organs, such as improving schizophrenia, autism - related psychiatric and psy-chological symptoms. Oxytocin exhibits its physiological func-tions by binding to its receptor (oxytocin receptor,OTR). Cur-
rently researchers are manipulating OT system by developing new OTR ligand ( agonists and antagonists ) , hoping to prevent and treat OTR related diseases. This paper reviews the latest devel-opment of OTR agonists, antagonists and its physiological roles in central nerve system and peripheral organs.

Objective To observe the cerebral edema occurrence ,evolution and prognosis of intra-arterial thrombolytic recanali-zation in patients with acute ischemic stroke .Methods From October 2010 to October 2012 ,36 patients who underwent the intra-arterial thrombolytic therapy were cerebral recanalization .They were randomly divided into two groups :1-3 h group and >3-6 h group .The alteration of brain edema was observed by cranial CT .Their mRS .NHISS and BI were scaled and recorded before and after intra-arterial thrombolytic therapy .Results After intra-arterial thrombolytic therapy ,the occurrence rate of cerebral edema was 94% ,appeared at the onset of more than 1 hours in all patients .Long-term follow-up showed ,cerebral edema location appeared obvious cerebromalacia .Cerebral edema and clinical outcome had not significant difference between 1-3 h group and 3-6 h group . Conclusion Cerebral edema and loss of cerebral tissue occurred almost inevitability in patients who received intra-artery thromboly-sis and recanalized their cerebral artery .It indicates that good local circulation and general condition may be favorable factors that can gradually reduce brain edema .