Objective To study the correlation between serum human cytomegalovirus (HCMV) DNA and peripheral T cell subsets as well as islet function in type 1 diabetic patients.Methods HCMV DNA levels in sera from 20 type 1 diabetic patients and 40 controls were measured by quantitative polymerase chain reaction (PCR),then the comparison and correlation analysis were made between HCMV DNA and T cell subsets,blood glucose (BG),insulin (Ins) and C peptide (C P).Results The positive rate and levels of HCMV DNA in type 1 diabetic patients were significantly higher than those of controls respectively.The percentage of CD 8 and the levels of fasting BG and BG 2 hours after meal in type 1 diabetic patients with positive HCMV DNA were by far higher than those in controls,while the percentage of CD 4,the ratio of CD 4/CD 8,the levels of fasting Ins,Ins and C P 2 hours after meal were significantly lower than those of controls.There existed the linear positive correlation between HCMV DNA and CD 8 or fasting BG,negative correlation between HCMV DNA and CD 4/CD 8,fasting Ins or fasting C P.Differences were not so significant between type 1 diabetic patients with negative HCMV DNA and controls compared with those between patients with positive HCMV DNA and controls.Conclusion HCMV infection is associated with dysfunction of islet and T cell mediated immunity in type 1 diabetic patients.

Objective To establish rat model of type 2 diabetes through a single subcutaneous injection of lipopolysaccharide which induced chronic inflammation.Methods The male Wistar rats(n = 30)were randomly divided control group (n = 10)and model group (n=20).Model group with LPS (300 μg·kg-1 ·day-1 )subcutaneous injection of eight weeks,rats in control group received isometric stroke-physiological saline solution injection in the same way.The changes in appearance,weight and fasting blood glucose (FBG)of rats were observed every week.At the end of the 8th week,thelevels of tumor necrosis factor a (TNF-α), interleukin-1(IL-1),interleukin-6 (IL-6),monocyte chemotactic protein-1(MCP-1)and fasting insulin(FINS)were measured.Oral glucose tolerance test (OGTT)and insulin release test(IRT)were also performed.The successful rat model was determined by the standards that FBG was ≥11.1 mmol/L.Results Model group rats reached the standard of type 2 diabetes after six weeks of LPS injection.Model group blood sugar is significantly higher than the control group (P <0.05).In addition,model group′s expression level of inflammatory cytokines in serum TNF-α,IL-1,IL-6,MCP-1 and FINS were significantly higher than control group (P <0.05).Oral glucose tolerance test,blood glucose levels higher than normal in model group,the insulin peak is lower than the normal group (P <0.05).Conclusion The success of establishing the animal model of type 2 diabetic rats which were injected of low dose of LPS by subcutaneous may be provide certain help for the etiology of diabetes research.

Objective Stem cell transplantation is a new approach to the treatment of lower limb ischemia ( LLI) .This study was to investigate the therapeutic effect of the transplantation of autologous bone marrow mesenchymal stem cells ( BM-MSCs) in the treatment of LLI. Methods We established the left LLI model in 12 New Zealand rabbits and divided them into a control and a trial group of equal number.The control animals were injected with DMEM, while the rabbits in the trial group with autologous BM-MSCs, into the ischemic skeletal muscle.Four weeks after injection, we performed CT angiography and perfusion imaging of both lower limbs of the rabbits and conducted HE staining of the paraffin sections of the skeletal muscle of the ischemic limbs. Results Dynamic ob-servation revealed different degrees of ecderon necrosis in 2 of the LLI models in the control group, even with toenail coloboma, but no necrosis in the trial group except for some slight muscular atrophy. Both collateral arteries and blood perfusion were obviously increased in the ischemic lower limbs in the trial than in the control group.HE stai-ning showed a significantly higher density and percentage of capillaries in the skeletal muscle fibers in the former than in the latter ([6.500 ± 1.049]/HP vs [3.670 ±0.816]/HP, [9.68 ±0.56]%vs [5.87 ± 0.86]%, P<0.01), with no necrosis in either group, nor hematoma, bony tissue, or fibroid tumor in the trial group. Conclusion Autologous transplantation of BM-MSCs can improve neovascularization in ischemic lower limbs in rabbits and can be used as a safe and effective treatment of limb ischemia.

Objective To discuss the nursing intervention of Rey syndrome in children patients.so as to search effective nursing measures.Methods 12 children patients with Rey syndrome received comprehensive nursing treatment with reducing intracranial pressure and were under continuous close observation.Slightest changes were feeded back timely,then effective nursing measures were taken to stabilize their illness,patients also received dietary therapy,medication nursing and rehabilitation training,etc.Results All patients ameliorated after 3 to 7 days,and achieved clinical recovery after 20 to 30 days without sequelae.Conclusions Early diagnosis and correct effective nursing care can improve curative rate and avoid the occurrence of complications and sequelae.

Objective To study the effects of Ganoderma lucidum polysaccharide (GLP) on PBMCs and the related immune mechanism.Methods PBMCs from cancer patients and healthy donors were isolated and treated with different doses of Ganoderma lucidum polysaccharides (10 ng/ml,50 ng/ml and 100 ng/ml).DC cell costimulatory molecules (HLA-DR,CD83,and CD11 c),Th1 (CD3 + CD8-IFN-γ+) cells,Th2 (CD3 + CD8-IL-4+) cells and NK (CD3-CD56+) were analyzed by FCM.Furthermore,The CD3+ CD4+ Th ceils were separated by immunomagnetic beads and stimulated with Ganoderma lucidum polysaccharides at different concentrations in culture.After 24 h,the cytokine expression levels of Th1 and Th2were detected by RT-PCR.The expressions of Th1 differentiation-related transcription factor,STAT4,were analyzed by Western blot.Results Ganoderma lucidum polysaccharides can significantly stimulate in vitro Thl cell differentiation (P＜0.01) in a dose depend manner.It correlates with an increased expression of STAT4 and the elevated mRNA expression levels of Th1 cytokineincluding IL-12,IFN-γ and TNF-α (P＜0.01).Conclusion Ganoderma lucidum polysaccharides may promote Th1 differentiation and increase the secretion of Th1 cvtokines through the upregulation of STAT4.

Objective To investigate the correlation between the expressions of vascular endothelial growth factor C (VEGF C) and vascular endothelial growth factor D (VEGF D) and the lymph node metastasis of groin and retroperitoneum in human epithelial ovarian carcinoma. Methods The expressions of VEGF C and VEGF D in benign, borderline, and malignant epithelial ovarian tumors were detected by immunohistochemistry and light microscopy. Results The expressions of VEGF C and VEGF D were found in benign, borderline, and malignant epithelial ovarian tumors. The expressions of VEGF C and VEGF D in ovarian carcinomas were significantly higher than those in benign and borderline ovarian tumors. The expression levels of VEGF C and VEGF D were correlated with peritoneal metastasis, lymph node metastasis of groin and retroperitoneum, and clinical stage, but not with distant metastasis, histology type, histological grade, and age. Conclusion Up regulation of VEGF C and VEGF D in epithelial ovarian carcinoma is advantageous to lymph node metastasis. This process may probably be correlated with lymphangiogenesis in tumors.

The roles of NF-kappaB (NF-kappaB) expression, Bax activity and cytochrome C (Cyt C) release, apoptosis of islet cells induced by high concentration glucose were explored in vitro. Pancreatic islet cells, which were isolated from Kunming mice, were cultured with different concentrations of glucose in DMEM, and divided into the following groups: G1, G2, G3, G4, G5, and G6 groups, corresponding to the glucose concentrations of 5.6, 7.8, 11.1, 16.7, 22.5, and 27.6 mmol/L, respectively. After culture for 120 h, insulin secretion was evaluated by radioimmunoassay, and the NF-kappaB expression was detected by immunocytochemistry. Bax activity and Cyt C release were measured by immunofluorescence, and apoptosis was examined by Hoechst33342 assay. The results showed that in G1, G2 and G3 groups, insulin secretion was enhanced with the increase of glucose concentration, and the NF-kappaB expression was also increased (P<0.05), but Bax activity, Cyt C release and apoptosis rate showed no significant difference among them. However, in G4, G5, and G6 groups, apoptosis rate of islet cells, NF-kappaB expression, Bax activity, and Cyt C release were all significantly increased, and insulin secretion was impaired as compared with G1, G2, and G3 groups (P<0.05). It was concluded that the exposure of islet cells to high glucose could induce islet cells apoptosis as well as impaired insulin secretion. The NF-kappaB signaling pathway and mitochondria pathway in islet cells might play some roles in the progressive loss of islet cells in diabetes. The inhibition of the NF-kappaB expression could be an effective strategy for protecting pancreatic islet cells.

To evaluate the prevalence of mosquito-borne viruses in Manshi and Ruili (Yunnan Province, China), we collected 2 149 mosquitoes (17 species) in August 2010. Virus isolation was undertaken by the cul- ture of baby hamster kidney cells (BHK-21 cells). Two virus-like isolates were obtained: DHL10M117 was isolated from collected in Mangshi; DHL10M110 was obtained from Anopheles vagus collected in Rui- li. Both isolates caused cytopathic effects,illness and death in suckling mice inoculated with these isolates via the intracerebral route. Two positive amplicons, 702-bp from the S segment and 456-bp from the M segment,were obtained using reverse transcription-polymerase chain reaction using primers specific for the Akabane virus (AKV). Phylogenetic analysis suggested that these two virus stains had a distant relation- ship with AKVs from Kenya and Australia,but were genetically close to those from Japan,South Korea, and Taiwan. However,they were separate from other Asian strains and grouped into a small branch. The highest nucleotide and amino-acid sequence identity of the S segment was found with the CY-77 strain from Taiwan (96.6% and 99.6% for DHL10M117 and 96.7% and 100% for DHL10M110,respectively). Com- parison of the M segment showed they shared the highest amino acid identity with CY-77 (99.6% and 100%, respectively), whereas the highest nucleotide identity was found with the Iriki strain from Japan (99.6% and 100%, respectively). Compared with the MP496 strain from Kenya,they displayed lower lev- els of sequence homology, at 69.7% and 70.0% for nucleotide sequences of the two loci,and 91. 0% for a- mino acids. Our results identified that DHL10M117 and DHL10M110 were strains of AKV,and provided molecular biological evidence for the existence of AKV in Yunnan Province. These AKV strains that are circulating in Yunnan Province share a close genetic relationship with strains from the rest of Asia. Culex tritaeniorhynchus and Anopheles vagus may serve as transmission vectors.
Amino Acid Sequence ; Animals ; Anopheles ; virology ; Base Sequence ; Bunyaviridae Infections ; virology ; China ; Cricetinae ; Female ; Humans ; Insect Vectors ; virology ; Male ; Mice ; Orthobunyavirus ; classification ; genetics ; isolation & purification ; physiology ; Phylogeny ; Sequence Homology ; Viral Proteins ; chemistry ; genetics

Objective To study the effects of benzyl propionate nandrolone (BPN ) on the nicotinamide phosphoribosyl transferase (Nampt), insulin receptor substeate-2 (IRS-2 )and pancreatic duodenal homeobox-1 (PDX-1)expressions, cell cycle changes as well as insulin secretion in pancreatic islet cell NIT-1 lines, and to explore the influence of BPN in the Nampte xpression in NIT-1 cells and insulin signaling molecules in high glucose oxidation stress.Methods The NIT-1 cells were cultured with different concentrations (5.6,11.1,16.7,and 27.6 mmol·L-1)of glucose,then they were treated with 10 mg·L-1 BPN for 48 h with no BPN treatment as corresponding control groups.The expression levels of Nampt,IRS-2,and PDX-1 were tested by Western blotting assay.The changes of cell cycle were determined by FCM and the cell insulin secretion levels were measured with radioimmunoassay.Results Compared with corresponding control groups,the expression levels of Nampt,IRS-2, and PDX-1 proteins in the NIT-1 cells in various BPM groups were increased (P<0.05 or P<0.01).The G0/G1 phase arrest was relieved (P<0.01)when the cells was cultured in low glucose (5.6 mmol·L-1 )condition,and the G2/M block was remitted significantly in high glucose (27.6 mmol·L-1 )condition (P<0.01),furthermore, the cell insulin secretion was promoted compared with control groups except 1 1.1 mmol· L-1 glucose group (P<0.01).Conclusion BPN can promote the expression levels of Nampt,ISR-2 and PDX-1 proteins in NIT-1 cells. There is close relationship between the Nampt expression in NIT-1 cells and insulin signaling pathway and BPN prevents the cells from insulin resistance.

Objective To study the expressions of nicotinamide phosphoribosyl transferase (Nampt)in main energy metabolism organs (liver, pancreas, skeletal muscle, and kidney ) of the rats with type 2 diabetes mellitus (T2DM),and to explore the correlation between the expression and distribution of Nampt and the occurrence of diabetes.Methods The SD rat diabetes model was established by injecting with streptozotcin (STZ).The SD rats were randomly divided into diabetes group and control group. Immunohistochemical Envision staining assay was used to detect the distribution and protein expressions of Nampt in liver,pancreas,muscle,and kidney tissues of the rats,at the same time the blood glucose and serum insulin levels were also be detected. Results The blood glucose level of the rats in diabetes group was significantly higher than that in control group (P<0.01),and the fasting insulin level was lower than that in control group (P<0.01).The Nampt expression in the liver tissue of the rats in diabets group was significantly increased,which distributed near the hepatic sinus in diabetes rats,and the Nampt expression was also increased in skeletal muscle in which the whole cell was thick dying;the Nampt expression mainly distributed in the renal tubular epithelial cells. Compared with control group, the positive expression rates of Nampt in liver,skeletal muscle,and kidney tissues of the rats in diabets group were significantly increased (P<0.05 or P<0.01).There was nearly no Nampt expression in pancreas tissue of the rats in diabetes group and the Nampt expression level was significantly lower than that in control group (P<0.01). Conclusion The Nampt expressions are much different in main energy metabolic organs of the rats with diabetes.It is suggested that Nampt may be used as a specific indicative marker in the process of diabetes.