Objective To investigate the prophylactic treatment of cytomegalovirus (CMV) infection in renal transplant patients.Methods Fifty-five patients were divided into two groups. Group A (n=27) was administered with ganciclovir (5 mg/kg every day) intravenously for 30 days. Group B(n=28) was the control group. According to donor (D) or recipient (R) CMV antibody categories,each group had D+/R-,D-/R+,D+/R+ three subgroups. All patients were followed up prospectively for 6 months with measurements of CMV antibody,CMV-DNA antigen,acute rejection for monitoring activity of CMV infections.Results The rate of CMV infection and disease was similar in both groups. The delay between transplantation and CMV infection was significantly longer in the ganciclovir group than in the control group (70.1?14.9 vs 50.2?9.8,P

Objective To investigate the expression of vascular endothelial growth factor (VEGF), angiopoietin and their receptors (VEGFR and Tie2) in aging mice kidney and the possible roles in aging mice. Methods Mice were divided as follows: 4-month old group (n=6), 9-month old group (n=6), 12-month old group (n=6) and 20-month old group (n=6). Paraffin sections of the mice kidneys were stained by PAS. The density of glomerular microvascular was determined by renal perfusion with fluorescent dyes. The level of VEGF, VEGFR2 (Flk-1), Ang-1, Ang-2, Tie2 mRNA expression and protein abundance in kidney was determined by real-time PCR, immunochemistry, immunofluorescence and Western blot. Results Compared with other three groups, in the 20-manth old group, the glomerulosclerosis index (GSI) increased remarkbly (2.48±0.79 vs 0.53±0.19, 0.69±0.18, 1.50±0.70, P<0.05); the fluorescence intensity in glomeruli decreased (P<0.05). lmmunohistochemistry demonstrated that the TGF-131 level in the aging kidneys showed an increase trend in the glomerular tubulointerstitium, and especially in the glomeruli. Real-time PCR results revealed that compared with 4-month old group mice, the mRNA expression of VEGF, Flk-1, Ang-1, Ang-2, Tie2 of the other three groups decreased, the gene levels of VEGF, Flk-1, and Ang-2 fell about 90%, 50% and 80% (all P>0.05), and the gene levels of Ang-1 and Tie2 fell about 75% and 40% in 20-month-old group (all P<0.05). Western blot domonstrated that the protein abundace of VEGF, Flk-1, Ang-1, Ang-2, Tie2 also declined with aging, the protein level of VEGF, Flk-1, Ang-1, Ang-2 and Tie2 dropped by about 35%, 50%, 15%, 13% and 21% respectively in 20-month-old group as compared to 4-month-old group (all P<0.05). Expression of above 5 factors and glomerular fluorescence intensity were negatively correlated with Scr (P<0.05). Conclusions The mRNA expression and protein abundance of VEGF, Flk-1, Ang-1, Ang-2, Tie2 in mice kidneys decreases with aging. Angiogenesis regulatory factors may play important roles in aging progression of the mice kidney.

To study the expression of herpes simplex virus type 2 latency-associated transcript (LAT) open reading frame 1 (ORF1) and its anti-apoptosis function induced by actinomycin D in Vero cells. The recombinant plasmid pEGFP-ORF1 was constructed and transfected into Vero cells, and the expression of ORF1 was identified by RT-PCR. The changes of Vero cells morphology induced by actinomycin D were observed by fluorescence microscopy, Hochest33258 fluorescence staining. Cells viability was evaluated by MTT assay and cells apoptosis rate was detected by flow cytometry. Double digestion and sequencing confirmed the pEGFP-ORF1 was constructed successfully, RT-PCR showed that the target gene was highly expressed in Vero cells. Hochest33258 staining reaveals that Vero cells transfected with pEGFP-ORF1 and induced apoptosis by actinomycin D had no changes in morphology. MTT assay showed that the viabilities of Vero cells transfected with recombinant plasmid pEGFP-ORF1 and induced apoptosis by actinomycin D has no statistically significant difference compared with the untreated normal control group (P > 0.05), but remarkable higher than Vero cells transfected with empty plasmid pEGFP-C2 and induced apoptosis by actinomycin D, the difference was statistically significant (P < 0.05). Flow cytometry assay shows that the cells apoptosis rate had no significant difference between pEGFP-ORF1 group and the normal group, but the cells apoptosis rate ofpEGFP-ORF1 was lower than the pEGFP-C2 group. HSV-2 LAT ORF1 gene can be expressed in Vero cells and can protect Vero cells from apoptosis induced by actinomycin D.
Animals ; Apoptosis ; physiology ; Cercopithecus aethiops ; Dactinomycin ; Herpes Simplex Virus Protein Vmw65 ; genetics ; Herpesvirus 2, Human ; genetics ; Open Reading Frames ; genetics ; Promoter Regions, Genetic ; Transcription, Genetic ; Vero Cells ; Viral Proteins ; genetics ; Virus Activation ; Virus Latency ; genetics ; physiology

Objective　To ascertain the natural infection rate of hemorrhagic fever with renal syndrome virus (HFRSV) among its hosts and the type of the natural foci for providing some baseline data for the immigrant health and epidemic prevention of the Three-Gorge region. Methods　Epidemiological survey on the field was performed including epidemiological data collection, ecology of rodents and pathogen detection. HFRS virus antigen of hosts were detected by the direct immunofluorescent assay (DIFA) technique and determination of HFRSV-RNA by ISH were carried out from HFRSV-Ag-positive animals. Results　HFRSV-Ag-positive animals were found in 5 migration areas ie Baitao Town of Fuling Section, Wansheng Village of Fengjie County and Dachang Town of Wushan County. The positive hosts were as follows, Rattus Norvegicus, Apodemus agrarius, Anourusurex squamipes, Mus musculus and Rattus flavipectus. The positive rate of HFRSV in the mice of 5 migration areas were 19.4%, 17.0%, 14.0%, 13.7%, and 8.5% respectively. The results showed that the lung tissues of some hosts in all five migration areas were HFRSV-RNA-positive. Baitao Town and Peishi Town were attributed to mixture type epidemic areas while. Kangle Town, Wansheng Village and Dachang Town were domestic rats type epidemic areas. Conclusion　This study shows that the five migration areas are natural epidemic foci of HFRS. It is predicted that maximum risk of HFRS breakout or epidemic may take place after the completion of the San Xia Reservoir(the Three-Gorges Reservoir), which results from rodent moving toward higher land. Therefore, deratization and preventive measures for rat are important in migration areas.

Objective To investigate the role of endothelial bioreacter device in sepsis porcine model.Method Sepsis porcine model was induced gy established endotoxin (LPS,0.25 mg·kg~(-1)) in healthy hybrid swines. The animals were randomly divided(random number) into endothelial bioreactor device group(EBR group) and sham circulation group( Sham group)( n = 6, respectively). After the infusion of endotoxin, extracorporeal circulation was started with the blood flow of 30 mL/min. The blood went through the endothelial bioreactor, then went back to the body via internal jugular vein in the EBR group. The bioreactor with the same size and without endothelial cells(ECs) was used in the sham group. Hemodynamic variables, blood biochemistry, inflammatory markers, Endothelin-1(ET-11) and yon Willebrand Factor(vWF) were examined just before and every hour after the injection. When the survival time of the animals was recorded,the animals were sacrificed to calculate the lung injury score. The time-dependent hemodynamics and cytokine data were compared between groups by repeated measurement ANOVA .Student's t -test was used to analyze the survival time. Results The mean artetial blood pressure (MAP) remarkably decreased in both groups after LPS injection, while the decreasing rate in EBR group was significantly lower than that in control group after 2 hours( P < 0.05). The ET- 1 level in EBR group increased after a slight decrease at the beginning, while that in the sham group went on increasing(P<0.01). The vWF levels increased first, then returned to the baseline in the sixth hour in both groups, while the change in EBR group was significantly less than that in the sham group(P<0.05). The Lung Injury Score in EBR-treated group was significantly lower than that in the sham group(6.1 ± 0.9 vs. 8.2 ± 1. 0, P < 0.05). These physiologic and biochemical alterations were associated with a significant advantage to the survivals in the EBR group when compared with the control sham group(6.7 ± 1.32 vs. 5.2 ± 0.61 h, P < 0.01 ). Conclusions Timely intervention in endotoxin shock with EC therapy by using tissue-engineered bioreactor may improve cardiovascular performance and alter the natural course of this disease process, probably via modulating ioflammation and coagulation cascades.

It is not fully clear why there is a higher contribution of pluripotent stem cells (PSCs) to the chimera produced by injection of PSCs into 4-cell or 8-cell stage embryos compared with blastocyst injection. Here, we show that not only embryonic stem cells (ESCs) but also induced pluripotent stem cells (iPSCs) can generate F0 nearly 100% donor cell-derived mice by 4-cell stage embryo injection, and the approach has a "dose effect". Through an analysis of the PSC-secreted proteins, Activin A was found to impede epiblast (EPI) lineage development while promoting trophectoderm (TE) differentiation, resulting in replacement of the EPI lineage of host embryos with PSCs. Interestingly, the injection of ESCs into blastocysts cultured with Activin A (cultured from 4-cell stage to early blastocyst at E3.5) could increase the contribution of ESCs to the chimera. The results indicated that PSCs secrete protein Activin A to improve their EPI competency after injection into recipient embryos through influencing the development of mouse early embryos. This result is useful for optimizing the chimera production system and for a deep understanding of PSCs effects on early embryo development.
Activins ; metabolism ; Animals ; Cells, Cultured ; Embryonic Development ; Germ Layers ; metabolism ; Mice ; Pluripotent Stem Cells ; cytology ; metabolism