Amplification Effect of CdS Quantum Dots on Electrogenerated Chemiluminescence of Ru(bpy)32+and Its Application in Determination of Catechol / 分析化学

The effect of CdS quantum dots (QDs) on the electrochemiluminescence (ECL) signal of Ru(bpy)32+ was studied. It was found that CdS QDs could enhance the anodic ECL of Ru(bpy)32+ by 4 times. The sensitization mechanism was discussed and the influence factors including concentrations of Ru (bpy)32+ and CdS QDs, pH of solution and scan rate on ECL intensity were investigated. On the basis of quenching effect of catechol on the ECL signal of CdS QDs-Ru(bpy)32+,a system for sensitive determination of catechol was established with a detection limit of 5.5 nmol/L (S/N=3). This method was applied to the detection of catechol in tea sample with satisfactory results.

MiR-503 promotes wound healing of diabetic foot ulcer by targeting FBN1

Objective: To highlight the relationship between miR-503 and wound healing of diabetic foot ulcer (DFU). Methods: Microarray analysis was used to detect the dysregulated miRNAs between the DFU tissues and normal tissues. The expression of miR-503 in tissues and serum of patients with DFU was detected by qRT-PCR technique. Then, CCK-8 assay was applied to determine the cell proliferation. TUNEL assay was used for assessing the apoptosis of cells after treatment with miR-503. Possible correlation between miR-503 and fbillin1 (FBN1) was predicted according to data accessed on RNA22 website online, and was detected for confirmation by luciferase reporter assay. Results: Microarray analysis showed that miR- 503 was significantly decreased in the DFU tissues compared with normal tissues. While marked increase in the expression of miR-503 in tissues and serum of patients with DFU was confirmed by qRT-PCR technique. Then, CCK-8 assay indicated that transfection of miR- 503 mimic obviously accelerated the cell proliferation. However, TUNEL assays suggested that miR-503 mimic inhibited the apoptosis of cells to improve the survival of fibroblasts. Besides, miR-503 AMO played a role in fibroblasts of DFU tissues exactly countering to miR-503 mimic treatment. It was predicted that MiR-503 is a complementary to the FBN1 by RNA22. Besides, SiRNA-FBN1 promoted the proliferation, but brought down the apoptosis of fibroblasts. Conclusions: MiR-503 regulates the function of fibroblasts and wound healing of patients with DFU by targeting FBN1 directly which provids a novel and critical target for diagnosis and treatment of DFU.

MiR-503 promotes wound healing of diabetic foot ulcer by targeting FBN1

Objective:To highlight the relationship between miR-503 and wound healing of diabetic foot ulcer (DFU).Methods:Microarray analysis was used to detect the dysregulated miRNAs between the DFU tissues and normal tissues. The expression of miR-503 in tissues and serum of patients with DFU was detected by qRT-PCR technique. Then, CCK-8 assay was applied to determine the cell proliferation. TUNEL assay was used for assessing the apoptosis of cells after treatment with miR-503. Possible correlation between miR-503 and fbillin1 (FBN1) was predicted according to data accessed on RNA22 website online, and was detected for confirmation by luciferase reporter assay.Results:Microarray analysis showed that miR- 503 was significantly decreased in the DFU tissues compared with normal tissues. While marked increase in the expression of miR-503 in tissues and serum of patients with DFU was confirmed by qRT-PCR technique. Then, CCK-8 assay indicated that transfection of miR- 503 mimic obviously accelerated the cell proliferation. However, TUNEL assays suggested that miR-503 mimic inhibited the apoptosis of cells to improve the survival of fibroblasts. Besides, miR-503 AMO played a role in fibroblasts of DFU tissues exactly countering to miR-503 mimic treatment. It was predicted that MiR-503 is a complementary to the FBN1 by RNA22. Besides, SiRNA-FBN1 promoted the proliferation, but brought down the apoptosis of fibroblasts.Conclusions:MiR-503 regulates the function of fibroblasts and wound healing of patients with DFU by targeting FBN1 directly which provids a novel and critical target for diagnosis and treatment of DFU.

3D visualization assisted proximal femoral nail anti-rotation surgery in the treatment of elderly intertrochanteric fracture / 局解手术学杂志

Objective To investigate the application of 3 D visualization (3 DV) assisted proximal femoral nail anti-rotation (PFNA) surgery in the treatment of elderly intertrochanteric fracture (ITF).Methods Totally 11 cases of elderly unstable ITF were treated with 3DV assisted PFNA fixation surgery.All these patients were examined with multi-row spiral CT volume scan,and the anatomical data of Dicom was reconstructed through M3 D software.Computer aided design (CAD) was performed based on 3 D reconstruction data,including anatomical data measurement,fracture reduction simulation,and proximal femoral head nail (PFNA) implant parameter design.And the CAD data were showed by S-3DV system during operation to guide the operation.Time of operation,intra-operative blood lose,post-operative drainage volume,hospital stay time and Harris hip score were recorded and compared to the non-3DV assisted PFNA procedure.Results In 3 DV-PFNA surgery,there was no significant difference between the preoperative planned average PFNA size and the implanted average PFNA size (P ＞ 0.05).Compared to the non-3DV-PFNA surgery,3 DV-PFNA surgery has less intra-operative blood lose and post-operative drainage volume,shorter time of preoperative traction reduction and operation (P ＜ 0.05).Harris hip function score of 3 DV-PFNA surgery was better than the conventional surgery 1 week after operation,and there was no significant difference in VAS score I month after operation.The 11 cases treated with 3DV-PFNA technology were healed well,and the rate of complications was lower than the conventional surgery.Conclusion 3DV technique can show accuracy CAD model of ITF,which may play an important role to improve efficiency and accuracy of PFNA surgery.

Sequence analysis of varicella-zoster virus gE gene in varicella-zoster virus strains with different clades / 病毒学报

To analyze the gE gene sequence of varicella-zoster virus (VZV) strains of different clades and subclades currently circulating in China. Eighteen skin lesion fluid swabs or skin scab pieces from patients with chickenpox or shingles were obtained from Beijing, Changchun, Lhasa and Urumqi between December 2010 and June 2011. The genotype of the virus strains was determined by a group of single nucleotide polymorphism (SNP) located in 15 ORFs, and the full-length gE genes of 18 strains representing all the clades in the study was amplified by PCR and sequenced. In addition to the synonymous mutations and non-synonymous mutations that were reported in the literature, there were 3 novel non-synonymous mutations (C56T, C1109T, C917A) and 4 new synonymous mutations (C54T, T1075C, T816C, G279A) found in the 8 strains analyzed. We found the VZV strains of clade 5 in Xinjiang for the first time,and the genotypes of some VZV strains circulating in Chagnchun could not be determined by the present methods. The analysis of gE gene sequences,revealed a novel non-synonymous mutations in the e1 and c1 epitopes, corresponding to the amino acid change of serine to tyrosine.

The genotyping and molecular evolution of varicella-zoster virus / 病毒学报

Varicella-zoster virus (VZV, Human herpesvirus 3) is a member of the family Herpesviridae, and is classified as alpha-subfamily along with HSV-1 and HSV-2. VZV is the causative agent of chicken pox (varicella) mostly in children, after which it establishes latency in the sensory ganglia with the potential to reactivate at a later time to cause shingles (zoster). Increasing molecular epidemiological studies in recent years have been performed to monitor the mutations in VZV genome, discriminate vaccine virus from wild type virus, study the phylogeny of VZV strains throughout the world, and understand the evolution of the different clades of VZV. The progress has great impact on the fields of epidemiology, virology and bioinformatics. In this review, the currently available data concerning the geographic distribution and molecular evolution of VZV clades are discussed.

A murine model with murine cytomegalovirus infection resulting in colon inflammation after allogeneic skin transplantation / 病毒学报

<p><b>UNLABELLED</b>To provide a reliable animal model for study of human CMV disease in gastrointestinal track, we tried to infect with murine cytomegalovirus (MCMV) in mice that were received allogenetic skin transplantation under immunosuppression. (1) Skin transplantation was performed between 18 donor C57BL/6 mice and 72 recipient BALB/c mice. (2) All recipient mice were then given Cyclosporine at 12 mg/kg daily for 2 weeks by intraperitoneal injection. Mice were randomly divided into 3 groups. Two experimental groups were received MCMV-infected mouse embryonic fibroblasts (MEF) at 10(4) PFU and 10(5) PFU respectively, and the control group received MEF only. We observed any possibly pathophysiological behavior changes and recorded the changes in body weight. The mice were sacrificed at 5d, 9d, 14d, 21d post infection and colon tissue was collected for analysis.</p><p><b>RESULTS</b>Mice infected with MCMV at 10(5) PFU group showed anorexia, lethargy and degression in locomotor activity. This group of mice showed significant decrease in body weight than that of other groups. Colon tissues were collected 14 days after infection. Histological examination revealed that the mucous layer became thinner in the proximal colon and increased number of lymphoid follicles in distal colon in infected animals. The changes in the mucosal structure was most prominent in the group 10(5) PFU MCMV. Viral DNA was present in the colon by in situ hybridization for IE1 gene, and viral gB transcript was positive by RT-PCR. One of the viral major proteins, pp65, was widely distributed in the colon by immunohistochemistry. These data demonstrated that MCMV established infection in colon of the mice after allogenetic skin transplantation. Electron microscopy showed that there were herpes virus particles in the colon tissue.</p><p><b>CONCLUSION</b>Infection with MCMV in mouse after allogenetic skin transplantation by nasal cavity inoculation resulted in the pathological changes in colon tissue similar to that of inflammation in human colon. The small animal model of colon inflammation may provide a platform for further study of pathogenesis as well as medical intervention of HCMV involved inflammation of human bowel.</p>

Evaluation of riboflavin photochemical inactivation efficacy of virus in red blood cells by using animal models / 中国实验血液学杂志

This study was aimed to evaluate the efficacy of riboflavin photochemical inactivation of virus in red blood cells by using animal models. human cytomegalovirus (HCMV) plus red blood cells were used as indicator, 30 BALA/c mice were divided into the experimental group (n = 10), virus control group (n = 10), visible light control group (n = 5) and red blood cell control group (n = 5). Mice in experimental group were inoculated with red blood cells inactive by the riboflavin photochemical, mice in virus control group was injected with red blood cells without riboflavin photochemical inactivation treatment, and mice in light control group was infused with red blood cells irradiated by visible light, and mice in red blood cells control group was injected with normal red blood cells. The virus was isolated in vitro from mice of various groups, the HCMV UL83 gene was detected by PCR, the PP65 antigen was identified by indirect immunofluorescence. The results indicated that the virus isolation, PCR detection and indirect immunofluorescence identification all showed positive in virus control group and visible light control group, while the results of detection in experimental and red blood cell control groups were negative. It is concluded that riboflavin photochemical viral inactivation of red blood cells is effective.

Genetic analysis of clinical varicella-zoster virus isolates collected in China / 中华流行病学杂志

Objective To study the genetic characteristrics of vaccine and varicella-zoster virus(VZV)strains isolated from patients with chickenpox or zoster by molecular analysis.Methods SNP based VZV genetic characteristrics were analyzed in 19 VZV isolates using the restriction fragment length polymorphisms analysis of DNA fragments of the open reading frames 6,38,62 and sequence alignment of the open reading frames 1,31,51,62.Results All vaccine strains were revealed Alu Ⅰ~-Pst Ⅰ~-Sma Ⅰ~+ BssH Ⅱ~+ Nae Ⅰ~+,96% clinical isolates were revealed Alu Ⅰ~+ Pst Ⅰ ~+Sma Ⅰ BssHⅡ Nae Ⅰ~-,2% clinical isolates were revealed Alu Ⅰ~-Pst Ⅰ~-Sma Ⅰ~+ BssH Ⅱ~+,2%clinical isolates were revealed Alu Ⅰ~+ Pst Ⅰ~-Sma Ⅰ~-BssHⅡ Nae Ⅰ~-by restriction fragment length polymorphisms analysis and sequence alignment revealed the mutations also presented in this four vaccine strains.Conclusion Use the restriction fragment length polymorphisms analysis of DNA fragments of the open reading,frames 6 and 62 could be distinguished VZV wild-type strains and vaccine strain in clinical isolates in China.In order to find the adverse effect caused by vaccine from certain company's,analysis on the SNPs in ORFs 1,31,51 and 62 is needed.

Effect of intense pulsed light on transforming growth factor-beta1 mRNA expression in rat skin / 南方医科大学学报

<p><b>OBJECTIVE</b>To observe the effect of intense pulsed light (IPL) on transforming growth factor-beta1 mRNA (TGF-beta1 mRNA) expression in rat skin and explore the molecular mechanisms of photorejuvenation.</p><p><b>METHODS</b>Fifteen SD rats were exposed to IPL in 3 dermal regions with triple pulses (duration of 4, 5, and 6 ms) at the energy density of 34 J/cm2 and pulse delay of 20 or 25 ms. On days 1, 3, 5, 7, 15, and 30 after the treatment, skin specimens from the treated and non-treated areas were obtained to detect TGF-beta1 mRNA expression with in situ hybridization.</p><p><b>RESULTS</b>In the UPL-exposed skin areas, TGF-beta1 mRNA expression was detected in the epidermal keratinocytes and dermal cells 1 day after the exposure, reaching the highest expression level on day 7 followed by gradual decrement since day 15, and till day 30, only weak expression was found in the dermal cells. In the non-exposed regions, the cells remained negative for TGF-beta1 mRNA.</p><p><b>CONCLUSION</b>IPL can enhance TGF-beta1 mRNA expression in the skin, suggesting that TGF-beta1 plays an important role in dermal remodeling in photorejuvenation.</p>

Seroepidemiology of varicella-zoster virus infection measured by the fluorescent antibody to membrane antigen test / 中华流行病学杂志

Objective Adopting serology assay-fluorescent antibody to membrane antigen (FAMA) as "gold standard" , sero-prevalence status of varicella-zoster virus (VZV) infection was investigated, in Guangzhou. Methods FAMA test was established with VZV infected human embryo fibroblasts as antigens and fluorescein isothiocyanate(FITC) labeled goat anti-human IgG as the secondary antibody. Sensitivity and specificity of the assay were evaluated. The sero-prevalence of anti-VZV IgG in 592 serum specimens randomly collected from a clinical laboratory, was analyzed with FAMA. Results Data from FAMA test showed no cross-reaction with other Herpesviruses when it was used to detect VZV antibodies. The overall prevalence of VZV antibody was 76.52%. Age-specific prevalence rates of VZV antibody in different age groups as: 1-3, 4-6, 7-13, 14-19, 20-29, 30-39, 40-49, ≥50, were found to be 14.67%, 51.56%, 73.91%, 91.26%, 92.78%, 95.65%, 98.11% and 100%, respectively.The sero-prevalence of 1-3 age group appeared the lowest but rose sharply with the increase of age but showing no association with gender. Conclusion Our data indicated that VZV infection occurred in early childhood, in Guangzhou, suggesting that the primary recipients of VZV vaccine should be under the 1-3 age group. Additional subjects for vaccination would be children above 3 years old with no history of VZV infection, and serology test negative for VZV. The assay was validated by its excellent specificity and could be used as the first choice in the detection of protective antibodies against VZV infection.

Modified technique for correction of the orbital hypertelorism / 中华整形外科杂志

<p><b>OBJECTIVE</b>To explore the possibility of para-orbital soft-tissue expansion, before orbital osteotomy and medial translocation procedures with a combined intracranial-extracranial approach.</p><p><b>METHODS</b>Tissue expansion in the region of the zygomatic and temporal has been undergone for 3 weeks before the traditional intracranial-extracranial approach for orbital osteotomy and medial translocation in two patients. The healing between the orbital bone was studied with measurement of interorbital distance and three-dimensional CT.</p><p><b>RESULTS</b>The inter-orbital distance of the two patients decreased from 4.4 cm and 3.2 cm to 2.0 cm and 1.4 cm. The intercanthal distance decreased from 6.7 cm and 4.8 cm to 5.0 cm and 3.8 cm.</p><p><b>CONCLUSIONS</b>The para-orbital soft-tissue expansion technique may be an effective technique for the stability of the corrected interorbital distance in orbital hypertelorism.</p>

Immunoregulation effects in vitro of the xenoprotein in combination with recombinant human granulocyte-macrophage colony stimulating factor and bacillus Calmette-Guerin / 中国实验血液学杂志

This study was aimed to investigate the effects of xenogeneic antigen neu-Fc in combination with the recombinant human granulocyte-macrophage colony stimulating factor (GM-CSF) and Bacillus Calmette-Guerin (BCG) on the regulation of Th1 and Th2 immune response in vitro. The rat neu L2-S2 domain was engineered as a chimeric protein with human IgG Fc. The eukaryotic expression vector was constructed. The recombinant protein was stably expressed in CHO cells and purified by rProtein A Sepharose Fast Flow column. The recombinant protein was identified by SDS-PAGE and Western blot. Peripheral blood mononuclear cells (PBMNCs) were obtained by means of standard Ficoll separation from the blood of healthy donors. Neu-Fc-induced PBMNC proliferation was tested by MTT. The production of IL-12 and IL-10 was measured by ELISA. The results showed that the level of IL-12 decreased and IL-10 increased after PBMNCs were incubated with MCF-7 cultural supernatant. 10 nmol/L neu-Fc strongly induced the cell proliferation. Compared with neu-Fc or GM-CSF or BCG treatment alone, neu-Fc in combination with GM-CSF and BCG significantly stimulated IL-12 production and inhibited IL-10 production (p < 0.01). It is concluded that the neu-Fc can stimulate the proliferation activity of PBMNCs. neu-Fc, GM-CSF and BCG costimulation efficiently induces Th1 immune response.

CONSTRUCTION AND IDENTIFICATION OF HCMV cDNA EXPRESSING LIBRARY AND SCREENING OF pp65 POSITIVE CLONES / 微生物学通报

In order to provide effective tool for further studying of the function of HCMV genome, developing of molecular vaccine and diagnostic reagents. Extraction of HCMV mRNA from HF cell infected by HCMV AD169 strain for 96h was reverse transcripted into cDNA, then was cloned into EcoR I-digested lambda gt11. HCMV AD169 strain cDNA expressing library has been constructed after packaging. The volume and the recombination rate of the prime cDNA expressing libraries was 3.6?10 6 and 96%, 168 positive clones of HCMV were screened by immune blotting with anti-HCMV mouse convalescent sera, 34 positive clones were obtained by dot nucleic acid hybridization with DIG-labled HCMV pp65 gene probe. 2 positive clones were amplified by HCMV pp65 all length primer. The PCR product has been tested by southern-blotting.The PCR product was sequenced and was taken as homology comparison by DNASIS software,and the homology is 98%.To lay the foundation of furher cloning,expressing the pp65 gene,further studying of the function of the pp65 prodct.

Relationship between Expression of podocin mRNA and Oxidative Stress in Rats with Adriamycin-Induced Nephrotic Syndrome / 实用儿科临床杂志

Objective To explore the possible association of oxidative stress reaction with the mRNA expression of podocin in rats with adriarnycin(ADR)-induced nephrotic syndrome.Methods mRNA expression of podocin in renal cortex were investigated by in situ hybridization staining and semi-quantitative RT-PCR in ADR-induced nephrotic rats,the level of malondialdehyde(MDA), superoxide dismutase(SOD)and total antioxidative capacity(T-AOC) in renal cortex were measured, then the relationship between them was evaluated.Results In ADR rats, MDA increased at d7 and reached a significant higher level at d14 and d21;SOD decreased at d14 and persisted to d21; T-AOC decreased at d21.The podocin mRNA mostly expressed in cytoplasm of glomerular cells. Both the number and the intensity of positive cells increased notably as time progressed in ADR rats.The level of podocin mRNA expression showed no obvious changes at d7, while prominently increased at d14 and pesisted to d21 in ADR rats.There was significant correlativity between the mRNA expression of podocin and the level of MDA,SOD,T-AOC.Conclusion The mRNA expression of podocin is correlated with oxidative stress reaction in rats with ADR-induced nephrotic syndrome.

Application of Polymerase Chain Reaction in Checking Human Cytomegalovirus o f Cerebral Palsy Children / 实用儿科临床杂志

Objective To explore the clinical application and significance of polymerase chain reactio n(PCR) in checking human cytomegalovirus (HCMV)in cerebral palsy children. Methods Collecting urine and serum of 56 cerebral palsy (CP) children,using PCR to detec t CMV DNA from urine,isolate CMV from urine,and indirect enzyme-linked immuno so rbent assay(ELISA) detecting CMV IgM、IgG of serum.Results In 56 cases,53.6%cases were CMV DNA positive,there were 9 cases CMV isolation,o bserving CMV characteristic cytopathic effect (CPE) and the positive value of se rum CMV IgM、IgG was 12.5%,37.5% respectively.The positive value in isolation o f the virus and CMV IgM was 100%,10% corresponding with that of CMV DNA.Comp ared the 2 former with the latter,it was significant(P0.05).Conclusions Using PCR can detect CMV DNA from CP children with CMV infection quickly.It can apply in detecting CMV in CP and provide credible evidence for intervention as f ar as early in children with CP. J Appl Clin Pediatr,2005,20(2):157-159