0.98); immature cells would display in the WBC histogram when in higher proportion. Conclusions The analyzer can be used to test blood cell parameters accurately and reliably. Its main performance indices accorded with the experimental requirements; The results were credible. It is necessary to checked with microscopy for DC before reported.

Objective To observe the protective effect of asiatic acid on hyperlipidemia golden hamsters induced by high fat diet and explore its mechanism.Methods Hyperlipidemic golden hamsters fed with high-fat diet were administered orally with asiatic acid (8,16,32 mg/kg)for 4 weeks.Levels of serum lipid content,liver histology,hepatic GSH-PX and SOD levels,serum ALT and AST activities were evaluated in golden hamsters,fluorescence quantitative polymerase chain reaction(RT-PCR)is using to examine the liver lecithin cholesterol fatty acyl transferase(LCAT) and class B typeⅠscavenger receptor(SR-BⅠ)mRNA expression. Results Compared with model group,the levels of TC,TG,LDL-C,TC/HDL,ALT and AST in low and mediate asiatic acid groups were all significantly decreased but the levels of SOD and GSH-PX were significantly increased(P<0.01).HE staining results showed that fat deposition in the liver were improved by administration of asiatic acid,and the expression of LCAT and SR-BⅠmRNA were increased.Conclusion Asiatic acid can effectively reduce blood lipid levels,and alleviate liver damage of the hyperlipidemia golden hamsters by lipid regulation and antioxidative ability augmentation.The increased levels of LCAT and SR-BⅠmRNA expression maybe involved in the mechanism of hypolipidaemic effect of asiatic acid.

Objective: To analyze the survival rate of stomach cancer patients and its variation during different periods in Linzhou city, Henan Province, from 1988 to 2004, and evaluate the level of secondary prevention and diagnosis of stomach cancer in this area. Methods: All of the incidence and death records of stomach cancer from 1988 to 2004 were collected and matched from Linzhou Cancer Registry. The records that were identified as duplicate cases or had only death certificate (DCO) were excluded. The tumor cause eliminated life tables in this area were calculated and linked to the data of incidence and death of stomach cancer. FivE-year observed survival rates and fivE-year relative survival rates in three periods (1990-1994, 1995-1999, and 2000-2004) were calculated using period survival analysis mehod. The relative survival curves in the three periods were plotted. Results:The 5-year relative survival rate of stomach cancer was 26.66% during 1990-1994, 32.01% during 1995-1999, and 40.43% during 2000-2004 in Linzhou city. It showed a gradually increasing trend. The 5-year survival rates were higher in males than those in females. During 1990-1994 and 1995-1999, the 5-year survival rates of gastric cardia cancer were higher than those of non-cardia cancer. During 2000-2004 period, the 5-year survival rate of gastric cardia cancer was lower than that of non-cardia cancer. Conclusion: The survival rates of stomach cancer in Linzhou city are increasing gradually since 1990s in 20 century. It indicates that the levels of secondary prevention and clinical diagnosis and treatment on stomach cancer kept increasing in this area.

Objective To construct an eukaryotic expression vector for TAP genes fused with enhanced green fluorescent protein(EGFP)gene,and to analyze the expression and subcellular localization of the fusion protein in A375 human malignant melanoma cells transfected with the eukaryotic expression vector.Methods A375 cells were cotransfected with the combination of plasmid(P)TAP1-EGFP or pTAP2-EGFP and pDsRed2-endoplasmic reticulum(ER),or with pEGFP-TAP1 and-TAP2,or monotransfected with pTAP1-EGFP or pTAP2-EGFP alone.The monoclonal A375 cells stably expressing TAP were obtained by G418 selection.Then.the distribution and expression of fusion proteins were assessed in A375 cells by using fluorescence microscopy and Western blot,respectively.Flow cytometry was used to measure the expression of HLA class Ⅰ on A375 cells.Results Transfection of A375 cells with pTAP1-EGFP or pTAP1-EGFP and pTAP2-EGFP significantly increased the expression of TAP 1 and TAP 2 in as well as HLA class Ⅰ antigen on A375 cells.The green fluorescence of TAP1-EGFP and TAP2-EGFP overlapped with the red fluorescence of ER marker in cotransfected cells.indicating that TAP was localized subcellularly on the ER.Conclusions The expression vector for TAP-EGFP fusion gene has been constructed cuccessfully and expressed in A375 cells,and the expressed fusion protein is subcelluiarly localized to ER.This study will provide a basis for the research into subsequent immune response following induction of TAP expression.

Objective To investigate the expression of glucose transporter type 1(GLUT-1)and hypoxia-inducible factor 1 (HIF-1)α in psoriatic lesions,and to explore their correlations with keratinocyte proliferation.Methods Biopsy specimens were obtained from 30 patients with psoriasis and 20 normal human controls.Immunohistochemistry and Western blotting were used to examine the protein expression of GLUT-1 and HIF-1α in these specimens.Results GLUT-1 and HIF-1α were mainly expressed in the basal layer of the control skin,but throughout the whole epidermis of psoriatic lesions.A significant increase was observed in the expression of GLUT-1 and HIF-1α in psoriatic lesions compared with that in the control skin (botb P＜0.01).In the case of psoriatic lesions,both the expression of GLUT-1 and HIF-1α was positively correlated with that of Ki-67(r=0.70,0.81 respectively,both P＜0.01),and positive correlation was also found between the expression of GLUT-1 and HIF-1α(r=0.85.P＜0.01).Conclusion Our data suggest that uprcgulation Of GLUT-1 and HIF-1α expression in psoriatic lesions might contribute to the proliferation of keratinocytes and psoriasis development.

To observe a PPAR-alpha agonist effect of N-oleoylethanolamine (OEA) on CB2 (cannabinoid receptor 2), an anti-inflammatory receptor in vascular endothelial cell, healthy HUVECs and TNF-alpha induced HUVECs were used to establish a human vascular endothelial cell inflammatory model. Different doses of OEA (10, 50 and 100 micromol x L(-1)) had been given to HUVECs, cultured at 37 degrees C for 7 h and then collected the total protein and total mRNA. CB2 protein expression was detected by Western blotting and CB2 mRNA expression was assayed by real-time PCR. As the results shown, OEA (10 and 50 micromol x L(-1)) could induce the CB2 protein and mRNA expression, but not 100 micromol x L(-1). To detect if anti-inflammation effect of OEA is partly through CB2, CB2 inhibitor AM630 was used to inhibit HUVEC CB2 expression, then the VCAM-1 expression induced by TNF-alpha was detected, or THP-1 adhere to TNF-alpha induced HUVECs was examined. OEA (50 micromol x L(-1)) could inhibit TNF-alpha induced VCAM-1 expression and THP-1 adhere to HUVECs, these effects could be partly inhibited by a CB2 inhibitor AM630. The anti-inflammation effect of OEA is induced by PPAR-alpha and CB2, suggesting that CB2 signaling could be a target for anti-atherosclerosis, OEA have wide effect in anti-inflammation, it may have better therapeutic potential in anti-inflammation in HUVECs, thus achieving anti-atherosclerosis effect.

Objective To study the effect of Jade -Screen Powder (JSP)on regulating expression of 5 microRNAs associated with helper T cells in asthmatic mouse model.Methods Forty Balb /c mice were randomly di-vided into 4 groups,1 0 mice for each group,namely normal control,asthma model,JSP treatment and Dexamethasone treatment.The mouse models of allergic inflammation on both upper and lower airways were established by ovalbumin sensitization and challenge.Interleukin(IL)-1 3 and IL -1 7 expressions were detected from lung homogenates by ELISA.Hematoxylin and eosin staining was also performed to observe the pathological changes in the lung tissue.The expressions of miR -1 46a,miR -1 46b,miR -21 0,miR -1 26 and miR -21 a were detected by quantitative real time PCR from splenocytes.Results The lower levels of IL -1 3 [(6.382 ±1 .690)μg/L]and IL -1 7 [(24.21 2 ± 1 .250)μg/L]were found in JSP treatment group compared with those in the asthma model group [(20.1 54 ±7.960)μg/L;(50.31 2 ±5.770)μg/L,rseparately],there was significant difference in IL -1 3 between JSP group and the asthma model group,as well as IL -1 7 (t =3.785,P =0.005;t =9.891 ,P =0.000).Same findings were found in Dexamethasone treated group as well [IL -1 3:(9.366 ±3.460)μg/L,IL -1 7:(29.1 32 ±4.960)μg/L;t =2.779, P =0.024;t =6.225,P =0.000].However,upregulation of miR -21 0 was observed in JSP treatment group (2.052 ± 0.871 )compared with that in the asthma model group (4.034 ±1 .379)(3.95 folds,t =2.71 8,P =0.026).Mean-time,the expression of miR -1 26 in JSP group (4.920 ±0.924)and Dexamethasone group (3.862 ±1 .51 0)in-creased compared with asthma model group (6.024 ±0.447)(2.1 5 folds,t =2.405,P =0.043,and 4.48 folds,t =-3.069,P =0.01 5).Conclusions Th2 and Th1 7 T cells participate in the pathogenesis of asthma and the asthmatic process can be inhibited by JSP.JSP may affect the helper T cells by regulating miR -21 0 and miR -1 26.

Objective This study was to investigate the characteristics of posterior corneal astigmatism (PCA) and aberration in cataract patients with high myopia.Methods A retrospective study was designed.Two hundred and eighty-two eligible eyes of 190 cataract patients were enrolled in Eye and ENT Hospital of Fudan University from September to December,2014.The eyes were classified into two groups according to axial length (AL):high myopia group with 139 eyes (AL≥26 mm) and control group with 143 eyes (AL was 20 to 25 mm).The mean keratometric mid-radius of curvature (Km),corneal central thickness (CCT),astigmatism and aberrations were measured by the rotating Scheimpflug System (Pentacam),and the AL were measured by the partial coherence interferometry (IOL Master).This study followed the Helsinki declaration,and was approved by the Ethic Committee of Eye and ENT Hospital,Fudan University.Informed consent was signed from each patient.Results In high myopia group,the mean PCA was 0.3 D (range 0 ～ 0.9 D) and 92.8％ eyes had PCA values ＜0.5 D.The steep corneal meridian was aligned vertically (60°～ 120°) in 87.1％ eyes for the posterior corneal surface.There was no significant difference in PCA between the high myopia group and the control group (P =0.797).Significant positive linear correlations was found between PCA and anterior corneal astigmatism (ACA),PCA and anterior corneal root mean square (RMS),PCA and anterior lower-order RMS,PCA and posterior corneal RMS,PCA and posterior high-order RMS,PCA and posterior lower-order RMS (r =0.235,P =0.005;r =0.217,P =0.010;r =0.229,P =0.007;r =0.395,P =0.000;r =0.243,P =0.004;r =0.384,P =0.000).Compared with total corneal astigmatism (TCA),anterior corneal measurements overestimated with-the-rule astigmatism (WTR) by a mean of (0.27 ± 0.18) D in 65.67％ eyes,underestimated against-the-rule astigmatism (ATR) by (0.27 ± 0.18) D in 88.10％ eyes and underestimated oblique astigmatism (Obl) by (0.22 ± 0.10) D in 63.33％ eyes.Compared with total corneal aberrations,anterior corneal aberrations measurements overestimated by (0.275 ±0.176) μm in 87.05 ％ eyes,and the anterior corneal astigmatism types had no effect on the result.Conclusions In high myopia group,92.8％ eyes had PCA values ＜0.5 D and the main astigmatism type in posterior corneal surface was ATR.The posterior corneal astigmatism and aberration were needed to consider in choosing intraocular lens (IOL) before cataract surgery.

The present study evaluated the effect of non-thermal plasma on skin wound healing in BalB/c mice. Two 6-mm wounds along the both sides of the spine were created on the back of each mouse (n=80) by using a punch biopsy. The mice were assigned randomly into two groups, with 40 animals in each group: a non-thermal plasma group in which the mice were treated with the non-thermal plasma; a control group in which the mice were left to heal naturally. Wound healing was evaluated on postoperative days (POD) 4, 7, 10 and 14 (n=5 per group in each POD) by percentage of wound closure. The mice was euthanized on POD 1, 4, 7, 10, 14, 21, 28 and 35 (n=1 in each POD). The wounds were removed, routinely fixed, paraffin-embedded, sectioned and HE-stained. A modified scoring system was used to evaluate the wounds. The results showed that acute inflammation peaked on POD 4 in non-thermal plasma group, earlier than in control group in which acute inflammation reached a peak on POD 7, and the acute inflammation scores were much lower in non-thermal group than in control group on POD 7 (P<0.05). The amount of granular tissue was greater on POD 4 and 7 in non-thermal group than in control group (P<0.05). The re-epithelialization score and the neovasularization score were increased significantly in non-thermal group when compared with control group on POD 7 and 10 (P<0.05 for all). The count of bacterial colonies was 10(3) CFU/mL on POD 4 and <20 CFU/mL on POD 7, significantly lower than that in control group (10(9) CFU/mL on POD 4 and >10(12) CFU/mL on the POD 7) (P<0.05). It was suggested that the non-thermal plasma facilitates the wound healing by suppressing bacterial colonization.

Background: African swine fever virus (ASFV), classical swine fever virus (CSFV), and porcine reproductive and respiratory syndrome virus (PRRSV) are still prevalent in many regions of China. Co-infections make it difficult to distinguish their clinical symptoms and pathological changes. Therefore, a rapid and specific method is needed for the differential detection of these pathogens. Objectives: The aim of this study was to develop a multiplex real-time quantitative reverse transcription polymerase chain reaction (multiplex qRT-PCR) for the simultaneous differential detection of ASFV, CSFV, and PRRSV. Methods: Three pairs of primers and TaqMan probes targeting the ASFV p72 gene, CSFV 5′untranslated region, and PRRSV ORF7 gene were designed. After optimizing the reaction conditions, including the annealing temperature, primer concentration, and probe concentration, multiplex qRT-PCR for simultaneous and differential detection of ASFV, CSFV, and PRRSV was developed. Subsequently, 1,143 clinical samples were detected to verify the practicality of the assay. Results: The multiplex qRT-PCR assay could specifically and simultaneously detect the ASFV, CSFV, and PRRSV with a detection limit of 1.78 × 10 0 copies for the ASFV, CSFV, and PRRSV, but could not amplify the other major porcine viruses, such as pseudorabies virus, porcine circovirus type 1 (PCV1), PCV2, PCV3, foot-and-mouth disease virus, porcine parvovirus, atypical porcine pestivirus, and Senecavirus A. The assay had good repeatability with coefficients of variation of intra- and inter-assay of less than 1.2%. Finally, the assay was used to detect 1,143 clinical samples to evaluate its practicality in the field. The positive rates of ASFV, CSFV, and PRRSV were 25.63%, 9.36%, and 17.50%, respectively. The co-infection rates of ASFV+CSFV, ASFV+PRRSV, CSFV+PRRSV, and ASFV+CSFV+PRRSV were 2.45%, 2.36%, 1.57%, and 0.17%, respectively. Conclusions The multiplex qRT-PCR developed in this study could provide a rapid, sensitive, specific diagnostic tool for the simultaneous and differential detection of ASFV, CSFV, and PRRSV.