Objective To understand laboratory diagnosis and clinical characteristics of human immunodeficiency virus (HIV ) infection and AIDS patients in three comprehensive hospitals .Methods Laboratory diagnosis and clinical characteristics dates of people living with HIV/AIDS patients consulted in Chongqing Emergency Medical Center between 2007 to 2013 were retrospective‐ly analyzed .Results Totally 47 355 cases were carried HIV antibody screening during 7 years ,179 cases of HIV antibody were positive in preliminary screening ,171 cases were confirmed as positive .Among 5 cases of HIV antibodies result unconfirmed ,2 ca‐ses were followed up ,1 case was ruled out HIV infection ,1 case was converted to HIV antibody positive .People living with HIV al‐ways merging double or multiple infection with hepatitis b virus (HBV) ,hepatitis c virus (HCV) and treponema pallidum (TP) and so on .People living with HIV aged from 18 to 86 years old ,9 .36% was over 60 years old .Most patient has two or more clinical manifestations when consulted a doctor .Conclusion There were false‐positive of HIV antibody preliminary screening ,HIV anti‐body positive results must be confirmed by Western blot confirmatory test .Uncertainty of HIV antibody results should be judged by regularly follow‐up or combining with other detection methods ,epidemiological data .Routine HIV antibody screening should be adopt for HBV ,HCV and TP infection .Elder patients should not be ignored .Clinical specificity of HIV/AIDS is not strong ,it is need to be valued and identified from other cause similar symptoms of diseases caused by phase identification ,in order to reduce missed diagnosis and misdiagnosis .

OBJECTIVEThis study aims to investigate the expression levels of serum and urinary vascular endothelial growth factor-A (VEGF-A) and epidermal growth factor-like domain 7 (EGFL7) in proliferating infantile hemangioma patients under propranolol treatment.

METHODSPropranolol (0.5-2 mg x kg(-1)) was orally administered to 30 infants every day for 4-8 months. The Achauer method was used to measure the tumor radius and thus evaluate the clinical curative effects of the treatment. Enzyme-linked immunosorbent assay was used to measure the serum and urinary concentrations of VEGF-A and EGFL7 at 0, 4, and 12 weeks after the treatment.

RESULTSThe treatment response was excellent in 2 patients, good in 11, moderate in 14, and poor in 3. Serum VEGF-A (335.692 pg x mL(-1) ± 136.146 pg x mL(-1)) was high before the treatment and then significantly decreased after 4 weeks (264.853 pg x mL(-1) ± 122.120 pg x mL(-1)) and 12 weeks (211.345 pg x mL(-1) ± 104.035 pg x mL(-1)) of treatment (P < 0.05). Urinary VEGF-A (76.234 pg x mL(-1) ± 24.169 pg x mL(-1)) was high before the treatment and then significantly decreased after four weeks (56.454 pg x mL(-1) ± l6.111 pg x mL(-1)) and twelve weeks (34.728 pg x mL(-1)) ± 12.656 pg x mL(-1)) of treatment (P < 0.05). Serum and urinary EGFL7 also decreased after the treatment, showing a positive relationship with VEGF-A.

CONCLUSIONPropranolol can be safely and effectively used to treat proliferating infantile hemangiomas. This treatment can reduce the peripheral serum and urinary concentrations of VEGF-A and EGFL7 in affected children.

Objective To discuss the changes of neutrophil gelatinase-associated lipocalin (NGAL) in patients with primary nephrotic syndrome (PNS)and the correlation with renal pathological type,renal tubulointerstitial lesions and the clinical indicators.Methods Forty patients with PNS were divided into acute kidney injury (AKI) group and non-AKI group according to whether renal tubular necrosis (ATN) occurred in renal pathology.Moreover,on the basis of pathological type they were divided into minimal change disease (MCD) group,mesangial proliferative glomerulonephritis (MsPGN) group,focal segmental glomerulosclerosis (FSGS) group,membrane proliferative glomerulonephritis (MPGN) group and membranous nephropathy (MN) group.Twenty healthy subjects and normal kidney tissues which came from 20 patients with renal tumor nephrectomy and were distant from the tumor sites were the control groups.Enzyme-linked immunosorbent assay (ELISA) was applied to detect the serum and urine level of NGAL,and immunohistochemical staining was used to observe the expression of NGAL in the renal tissue.Results (1)The serum and urine level of NGAL and the expression of NGAL in the renal tissue in the PNS complicated with AKI group were significantly higher than that in the PNS without AKI group and in the control group(P ＜ 0.05).(2)The serum and urine level of NGAL and the expression of NGAL in the renal tissue were enhanced in MPGN group and FSGS group than that in the other three groups(P ＜ 0.05).(3) Before developing to severe tubulointerstitial lesions,with the aggravation of tubulointerstitial damage,the serum and urine level of NGAL and the expression of NGAL in the renal tissue were increased.But when renal tubular interstitial lesions developed to severe disease,serum level of NGAL and the expression of NGAL in the renal tissue were decreased(P ＜ 0.05).(4)The serum and urine level of NGAL and the expression of NGAL in the renal tissue were positively correlated with serum creatinine(r values were 0.198,0.352,0.146 respectively,P values were 0.048,0.000,0.028 respectively),were positively correlated with blood urea nitrogen(r values were 0.199,0.278,0.325 respectively,P values were 0.043,0.000,0.019 respectively),were negatively correlated with serum albumin(r values were-0.384,-0.318,-0.259 respectively,P values were 0.028,0.024,0.020 respectively) and were negatively correlated with urine osmotic pressure(r values were-0.250,-0.256,-0.277 respectively,P values were 0.012,0.027,0.002 respectively).Conclusion NGAL is a sensitive biological parameter for predicting AKI in the patients with PNS,and it can be used to evaluate the degree of tubulointerstitial lesions and renal function to a certain extent.

Objective To evaluate the clinical efficacy of propranolol in treating proliferating infantile haemangiomas,and to measure the expression levels of vascular endothelial growth factor-A (VEGF-A) and hypoxiainducible factor 1α (HIF-1α) in sera and urine of patients during the treatment.Methods Thirty infants with proliferating haemangiomas were treated with propranolol at doses of 0.5-2 mg/kg per day.The radius of haemangiomas was measured,and blood and urine samples were obtained from these patients before,and at 4 and 12 weeks after the beginning of treatment.Clinical efficacy was estimated according to a four-graded scale as well as the feedback from parents of these patients.Enzyme-linked immunosorbent assay (ELISA) was performed to determine the serum and urine concentrations of VEGF-A and HIF-1α.Thirty check-up infants collected from the Department of Child Health Care served as the healthy controls.Statistical analysis was done by two-way analysis of variance followed by the least significant difference (LSD) test.Results After 12 weeks of treatment,clinical response was excellent in 2 patients,good in 11,moderate in 14,and poor in 3.The serum levels of VEGF-A and HIF-1α were (268.174 ± 95.056) μg/L and (10.809 ± 1.686) mg/L respectively in the control group,sequentially decreased in the patients from baseline to 4 and 12 weeks after the beginning of treatment (VEGF-A:(385.692 ± 136.146) vs.(264.853 ± 122.12) vs.(211.345 ± 104.035) μg/L; HIF-1α:(31.462 ± 7.458) vs.(21.454 ± 5.489) vs.(12.052 ± 3.623) mg/L).The trend in expression changes of VEGF-A and HIF-1α in urine samples was similar to that in blood samples in these patients.Positive correlation was observed between the expression level of VEGF-A and HIF-1α in sera (r=0.730,P＜ 0.05) and urine (r=0.667,P＜ 0.05) of these patients.Moreover,the levels of serum VEGF-A,urine VEGF-A,serum HIF-1α and urine HIF-1α were all negatively correlated with the time course following propranolol administration (r =-0.390,-0.689,-0.806,-0.683,P ＜ 0.05,0.01,0.05,0.01 respectively).Conclusion Propranolol is effective for the treatment of proliferating infantile haemangiomas,likely by reducing serum and urinary concentrations of VEGF-A and HIF-lα in children.

Objective To investigate the role of silent mating type information regulation 2 homologue 1(SIRT1) in renal ischemia-reperfusion(IR) injury and its effect on NF-κBp65-peroxisome proliferator-activated receptor gamma coactivator 1 alpha (PGC-1α) signal pathway in mice.Methods Seventy-two healthy C57BL/6 male mice were randomly divided into four groups:control group(n=18),sham-operated group(n=18),IR group(n=18),resveratrol group(n=18).Bilateral renal pedicle were clamped for 45 min was adopted to establish the model of acute ischemic renal injury,to give 2％ dimethyl sulfoxide or resveratrol by intraperitoneal injection for 7 days before modeling.Determination techniques included routine biochemical methods for the the levels of Scr and BUN,spectrophotometry for the level of superoxide dismutase (SOD),HE staining for the histological changes as well as immunohistochemical method and Western blotting for the expressions of SIRT1,NF-κBp65 and PGC-1α,respectively.Results Compared with that in control and sham-operated groups,the levels of serum Scr and BUN were higher and SOD levels in renal tissues were lower at 12 h and 24 h after operation in IR groups(P ＜ 0.05).HE staining revealed evident pathological lesions including necrosis of renal tubular epithelial cells in IR group.Compared with that in IR group,resveratrol attenuated the above-mentioned changes.Western blotting revealed the up-regulated SIRT1 expression and the activated NF-κB signal pathway,the up-regulated p65 expression and the down-regulated PGC-1αexpression subsequent to IR(P ＜ 0.05).Both Western blotting and immunohistochemistry showed that the expressions of SIRT1 and PGC-1α in resveratrol group were up-regulated compared to that in IRgroup(P ＜ 0.05),while the NF-κBp65 expression in resveratrol group was down-regulated(P ＜ 0.05).Conclusions In mouse model of renal ischemia-reperfusion injury,the activation of SIRT1 can inhibit the NF-κBp65 expression and accordingly up-regulated PGC-1α level,contributing to inhibiting inflammatory reactions and attenuating oxidative stress-induced injury in the protection of the kidneys.

The objective of this study is to explore the application possibility of chitosan/pcDNA-EGFP-TGFPbeta1 nanoparticles in the transfection of synovial-derived mesenchymal stem cells (SDMSCs). Chitosan/pcDNA-EGFP-TGFbeta1 nanoparticles were fabricated through method of ionic crosslinking. The SDMSCs were harvested from rabbit joints and cultured to passage 3. The SDMSCs were then transfected with chitosan/pcDNA-EGFP-TGFbeta1 nanoparticles. Scanning electronic microscope (SEM) was employed to detect the shape and diameter of the nanoparticles. The transfected SDMSCs were examined under the fluorescence microscope and detected through the flow cytometry (FCM). The SEM examination showed that the contour of the fabricated chitosan/pcDNA-EGFP-TGFbeta1 nanoparticles was round and its average diameter was 50 nm. After being cultured for 48 h, the SDMSCs transfected by chitosan/pcDNA-EGFP-TGFbeta1 nanoparticles could be detected under the fluorescence microscope, and the live SDMSCs could also be examined through FCM. The transfection rate was 8% - 10%. Therefore, it suggested that the chitosan/pcDNA-EGFP-TGFbeta1 nanoparticles fabricated through the method of ionic crosslinking could transfect the SDMSCs, but the transfection rate should be improved.
Animals ; Chitosan ; chemistry ; Genetic Vectors ; Green Fluorescent Proteins ; genetics ; Mesenchymal Stromal Cells ; cytology ; Microscopy, Electron, Transmission ; Microscopy, Fluorescence ; Nanoparticles ; chemistry ; Rabbits ; Transfection ; Transforming Growth Factor beta1 ; genetics

BACKGROUND:Articular chondrocytes with the ability of autocrine and paracrine can provide the growth factors and microenvironment for synovial mesenchymal stem cels differentiating into the chondrocyte. The
three-dimensional scaffold could provide space for cels adhesion, proliferation and differentiation.
OBJECTIVE: To study the ability of chondrogenesis by co-culturing synovial mesenchymal stem cels and chondrocytes under the three-dimensional condition.
METHODS:The synovial membrane and articular cartilage were harvested from rat knee joint. The synovial
mesenchymal stem cels and chondrocytes were obtained through the method of enzyme digestion. The passage 3 synovial mesenchymal stem cels and passage 2 chondrocytes were co-cultured in the chitosan/I colagen
composite scaffolds at the ratio of 1:2. Then, the cels/scaffold composite was harvested to be examined
morphologicaly, histologicaly and immunohistochemicaly after being cultured 21 days. The confocal laser was also employed to detect the cels distribution in the scaffold.
RESULTS AND CONCLUSION: After being cultured 72 hours, it could be observed from the cels/scaffold composite examined through the scanning electron microscope that the cels adhered on the surface of the
scaffold and extracelular matrix surrounding the cels was seen on the scaffold. After being cultured 21 days, it could be found through the confocal laser scanning that the cels were wel-distributed on the scaffold, and cels decreased gradualy. Type II colagen was positive in the extracelular matrix immunohistochamicaly. It
suggested from this study that the synovial mesenchymal stem cels could be co-cultured with chondrocytes in the chitosan/I colagen composite scaffolds and have the ability of chondrogenesis differentiation.

BACKGROUND:Synovial mesenchymal stem cells have the ability of multilineage differentiation in vitro, which are expected to be seed cells for the treatment of cartilage defects in cartilage tissue engineering. Appropriate growth factors are critical for the chondrocyte differentiation of synovial mesenchymal stem cells.
OBJECTIVE:To study the role of secreted factors by chondrocytes to induce chondrogenesis of synovial mesenchymal stem cells.
METHODS:The synovial mesenchymal stem cells and chondrocytes were harvested from rat knee joints and cultured through the digestion method. The supernatant was col ected from chondrocytes, and centrifuged, filtered and cryopreserved. The third passage synovial mesenchymal stem cells centrifuged as pel ets were cultured in the chondrocyte supernatant for 21 days. And the cells morphology was examined and the type II col agen and aggrecan were detected through immunohistochemistry and RT-PCR.
RESULTS AND CONCLUSION:The synovial mesenchymal stem cellpel ets cultured in the chondrocyte supernatant became cartilage-like tissue after 21 days. The type II col agen was detected positively in the matrix of synovial mesenchymal stem cellpel et immunohistochemical y. RT-PCR examination showed that the type II col agen and aggrecan expressed in the synovial mesenchymal stem cellpel et cultured in the chondrocyte supernatant. It suggested that synovial mesenchymal stem cellcould be induced to differentiate into chondrocytes depending on soluble factors secreted by chondrocytes.

ObjectiveTo summarize the technique and preliminary outcome of Neuroform stent combined with Guglielmi detachable coil (GDC) to treat wide-necked intracranial aneurysms. Methods32 cases with aneurysms which underwent 32 endovascular procedures performed by using stent were retrospectively analyzed.The ratio of aneurysm neck/body is 1/2～1/1. Results24 aneurysms were completely occluded and other 8 were incompletely (>95%) occluded. Transient ischemia of cerebral occured in 2 cases. 14 aneurysms were followed up 0.5～1 year after. 2 aneurysms of them appeared neck remnant growth.ConclusionUsing Neuroform stent combined with GDC to treat wide-necked intracranial aneurysm may prevent the herniation of GDC into the artery and increase the outcome of wide-necked intacranial aneurysm.

BACKGROUND:Compared with other sources of mesenchymal stem cells, synovial-derived mesenchymal stem cells have significant characteristics of chondrogenesis and cloning. Therefore, synovial-derived mesenchymal stem cells are one of the most promising seed cells in cartilage tissue engineering.
OBJECTIVE:To isolate and culture synovial-derived mesenchymal stem cells of Sprague-Dawley rats, identify the multipotential differentiation and the potential ability of chondrogenic differentiation in three-dimensional culture condition.
METHODS:The synovium tissue was harvested from Sprague-Dawley rats. The synovial-derived mesenchymal stem cells were isolated with typeⅠcol agen enzyme digestion method and cultured in vitro. The passage 3 cells were detected with giemsa staining, the cellcycle, adipogenic and osteogenic differentiation were determined. The passage 3 cells were centrifuged as pel ets and cultured in the chondriogenic medium for 21 days. And the pel ets were examined by toluidine blue staining, typeⅡcol agen immunohistochemical staining and RT-PCR.
RESULTS AND CONCLUSION:The mesenchymal stem cells isolated from the synovium tissue of rats have the characteristics of mesenchymal stem cells, and exhibit fibroblast-like morphology after cultured in vitro. The multilineage differentiation potentials were also revealed. After the cellwere cultured in chondrogenic medium for 21 days, chondroid tissue was found, type II col agen and aggrecan could be detected positively by toluidine blue staining, typeⅡcol agen immunohistochemical staining, and expressed by RT-PCR examination. Therefore, synovial mesenchymal stem cells have a chondrogenic differentiation potential.