Loop-mediated isothermal amplification (LAMP) is a novel in vitro nucleic acid amplification method conducted under isothermal conditions with the advantages of high specificity, sensitivity, rapidity and easy detection. Since it was established in 2000, it has been widely applied in various fields of analytical science including the diagnosis of a variety of pathogens, identification of embryo sex, detection of genetically modified organisms and cancer gene identification. Additionally, significant progress has been made in the optimization of the LAMP method, such as accelerated reactions, simplified sample processing, the realization of multiplex amplification, and the enhanced specificity of reaction and detection methods. LAMP technology also shows much potential to be adopted as part of point-of-care testing platforms by the micromation, automation and integration with other technologies such as Lab-on-a-Chip and digital nucleic acid amplification. This review summarizes the latest advances in the LAMP technique and its applications in developing point-of-care testing platforms.
Diagnostic Techniques and Procedures ; trends ; Humans ; Nucleic Acid Amplification Techniques ; methods ; Point-of-Care Systems ; trends ; Virus Diseases ; diagnosis ; virology ; Viruses ; genetics ; isolation & purification

Objective To study the mechanism of miR?200c in regulating RMP7?induced increases of blood?tumor barrier(BTB)permeability by targeting Ras homolog gene family member A(RhoA). Methods Endogenous expression of miR?200c was detected by real?time PCR in hu?man cerebral microvascular endothelial cell line hCMEC/D3(ECs)after RMP7 treatment. miR?200c mimic and miR?200c inhibitor were transfect?ed into GECs(ECs with U87 glioma cells co?culturing),respectively. Transfection efficiency of miR?200c mimic and miR?200c inhibitor were de?termined by real?time PCR. HRP flux and TEER assays revealed BTB permeability. The protein expression level of RhoA was assessed by West?ern blotting. The distribution of RhoA was assessed by immunofluorescence microscopy. RhoA luciferase assays were performed using the Dual?Lucif?erase reporter assay system. Results RMP7 significantly induced a decrease in miR?200c expression in GECs of BTB. miR?200c mimic and miR?200c inhibitor were successfully transfected into GECs. Overexpression of miR?200c inhibited endothelial leakage and restored normal transendo?thelial electric resistance values. Simultaneously ,overexpression of miR?200c significantly reduced the protein expression level of RhoA. In addi?tion,immunofluorescence analysis revealed that the distribution of RhoA in the cytoplasm and nuclei of GECs were decreased in miR?200c mimic group. RhoA was one of the direct targets of miR?200c with the specific binding site being located at the seed sequence. The results of miR?200c si?lencing were opposite to that of the miR?200c overexpression group. Conclusion miRNA?200c regulated RMP7?induced increases in BTB perme?ability by targeting RhoA.

Arteriovenous Fistula ; therapy ; Child ; Embolization, Therapeutic ; Humans ; Male ; Pulmonary Artery ; abnormalities ; Pulmonary Veins ; abnormalities

Objective To explore the curative effect of endourological treatment of pediatric urethral hemangioma with holmium laser.Methods Two children with urethral hemangioma were enrolled in this study. One urethral hemangioma in the bulbous urethra,another in the posterior urethra. Modalities of diagnosis before operation included B ultrasound,intravenous urogram(IVU) and endoscopy. Two children with urethral hemangioma underwent holmium laser therapy.Results After endourological trearment with holmium laser, two children had been cured.Six months to 4 years follow-up did not find recurrent urethral hemangioma.Conclusions The diagnosis of pediatric urethral hemangioma mainly rely on endoscopy. Endourological treatment with holmium laser is a minimally invasive,safe and effective methods for pediatric urethral hemangiomas.

Objective To investigate the expression of miR-29s in the glioma stem cells,and explore how the members of miR-29s affect the bio-logical behaviors of glioma stem cells. Methods Eight patient specimens were used to culture glioma stem cells. Real-time PCR was adopted to test the expression of miR-29s. CCK-8 analysis was performed to test the proliferation ,Transwell was used to test cell migration and invasion ,and flow-cytometry analysis was carried out to test apoptosis. Results miR-29a,miR-29b and miR-29c were decreased in glioma stem cells. Over-ex-pression of miR-29s could inhibit the proliferation,cell migration and invasion,but promote apoptosis of glioma stem cells. Conclusion miR-29s acts as a cancer suppressor gene in the glioma stem cells ,and miR-29a plays the dominant functional role in the family.

Objective To construct the prokaryotic vector and to express a novel candidate gene IDD01 associated with hepatocellular carcinoma (HCC) for its functional study. Methods The open reading frame (ORF) of IDD01, amplified from HCC tissue by RT PCR, was inserted into the expression vector pMAL c2X. The recombinant vector was transformed into E. coli TB1, and the expression products were analyzed by SDS PAGE. Results The length of PCR product was about 770 bp. The restriction enzyme digestion and sequencing conformed that the gene segment was correctly inserted into the vector. SDS PAGE and density scanning indicated that the protein was expressed as a fusion protein with 28 KD of molecular weight and the fusion protein possessed 30.4% of the total bacterial protein. Conclusion The recombinant vector is constructed successfully and IDD01 can be expressed at a high level, which lays a foundation for the further research on the functions of IDD01 gene.

Abstract： Objective To evaluate the filtration effects of various nanofiltration systems on intravenous human immunog⁃ lobulin（IVIG）in order to screen the optimal nanofiltration system. Methods Various nanofilters were used for IVIG filtration to determine the best one and then various prefilters were selected to combine with the optimal nanofilter for IVIG filtration to determine the optimal nanofiltration system. Results The tangential flow（cross flow）nanofilter showed better filtering effect than dead end（direct current）nanofilter，and nanofilter C was the best one. The effect of deep filtration prefilter was better than that of absolute filtration prefilter，and prefilter Y1 in series with nanofilter C was the optimal nanofiltration system. Conclusion The optimal nanofiltration system was determined through the effect evaluation of various nanofiltration systems filtering for IVIG.

OBJECTIVETo evaluate the efficacy and safety of testosterone undecanoate (TU) in the treatment of late-onset hypogonadism (LOH) by meta-analysis.

METHODSWe searched Pubmed (until April 1, 2014), Embase (until March 28, 2014), Cochrane Library (until April 17, 2014), CBM (from January 1, 2001 to February 2, 2014), CNKI (from January 1, 2001 to February 2, 2014), Wanfang Database (from January 1, 2000 to February 2, 2014), and VIP Database (from January 1, 2000 to Febru ary 2, 2014) for randomized controlled trials of TU for the treatment of LOH. We evaluated the quality of the identified literature and performed meta-analysis on the included studies using the Rveman5. 2 software.

RESULTSTotally, 14 studies were included after screening, which involved 1 686 cases. Compared with the placebo and blank control groups, TU treatment significantly increased the levels of serum total testosterone (SMD = 6.22, 95% CI 3.99 to 8.45, P < 0.05) and serum free testosterone (SMD = 4.35, 95% CI 1.86 to 6. 85, P < 0.05) but decreased the contents of luteinizing hormone (WMD = -2.23, 95% CI -4.03 to -0.42, P < 0.05), sex hormone binding globulin (WMD = 2.00, 95% CI 1.38 to 2.63, P < 0.05). TU also remarkably reduced the scores of Partial Androgen Deficiency of the Aging Males (WMD = -9.49, 95% CI -12.96 to -6.03, P < 0.05) and Aging Males Symptoms rating scale (WMD = -2.76, 95% CI -4.85 to -0.66, P <0.05) but increased the hemoglobin level (SMD = 2.35, 95% CI 0.29 to 4.41, P < 0.05) and packed-cell volume (SMD = 4.35, 95% CI 1.36 to 7.33, P < 0.05). However, no significant changes were shown in aspertate aminotransferase, alanine transaminase, prostate-specific antigen, or prostate volume after TU treatment (P > 0.05).

CONCLUSIONTU could significantly increase the serum testosterone level and improve the clinical symptoms of LOH patients without inducing serious adverse reactions. However, due to the limited number and relatively low quality of the included studies, the above conclusion could be cautiously applied to clinical practice.

Androgens ; therapeutic use ; Hemoglobin A ; metabolism ; Humans ; Hypogonadism ; blood ; drug therapy ; Luteinizing Hormone ; blood ; Male ; Prostate-Specific Antigen ; Randomized Controlled Trials as Topic ; Sex Hormone-Binding Globulin ; metabolism ; Testosterone ; adverse effects ; analogs & derivatives ; blood ; pharmacology

OBJECTIVETo investigate the effect of serum contained Chinese drugs for nourishing Shen and activating blood (S-NSAB) on activity of purified retinal ganglion cells (RGCs) cultured in high glucose medium.

METHODSPurified RGCs of SD rats were cultured in stimulative stable high glucose (50 mmol/L) condition (SHG) and fluctuated glucose condition (FGC) separately, they were intervened with S-NSAB, and the lactate dehydrogenase (LDH) leakage was detected by spectrophotometer for estimating the activity of RGCs.

RESULTSLDH leakage (U/L) in SHG culture was 1 349.17 +/- 215.50 at 24 h, 1220.24 +/- 124.53 at 48 h and 1982.14 +/- 219.03 at 72 h, all significantly lower than that in normal control at the corresponding time points (1628.10 +/-122.10, 1484.13 +/- 127.55 and 2155.75 +/- 140.44, respectively, P<0.05), whereas it was obviously higher in FGC culture at 72 h (2299.60 +/- 88.35), showing that LDH leakage in FGC was significantly higher than that in SHG at the corresponding time points (P<0.05). The LDH leakage was obviously decreased by Chinese medicine intervention with S-NSAB both in SHG at 72 h (1797.62 +/- 146.40) and in FGC at 48 h (1259.92 +/- 87.74) and 72 h (1940.40 +/- 155.47), the difference between pre- and post-intervention was significant (P<0.05).

CONCLUSIONFluctuated glucose conditions of culture medium could obviously damage the membranous stability of RGCs to enhance their permeability and lower the activity of cells; S-NSAB could improve these abnormalities in either SHG or FGC condition, which may be one of the important mechanisms of Chinese formula for nourishing Shen and activating blood in preventing and treating diabetic retinopathy.

Animals ; Animals, Newborn ; Cell Survival ; drug effects ; Cells, Cultured ; Culture Media ; pharmacology ; Drugs, Chinese Herbal ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Retinal Ganglion Cells ; cytology ; drug effects ; Serum

Objective To establish the orthotopic human pancreatic cancer model in mice and study the method to detect the tumor growth. Methods Human pancreatic cancer cell lines SW1990 in logarithmic phase was made into cell suspension and in situ injected into the envelope of pancreatic tail of BALB/c nude mice. High-frequency endoscopy ultrasound was used to observe the growth of tumor mass and its imaging characteristics were studied. Results 20 nude mice were successfully implanted, and 1 died 25 h after implantation. 14 days after implantation, the tumor of pancreatic cancer on EUS was (8.09 ± 2.61) mm3, the tumor appeared as homogeneous hypoechoic mass with clear boundary, and envelope as well as sound halo was present, and the shape was regular; there was low speed circular vasculature signal in around 30% of tumor.The tumor size increased to (12.40 ± 3.51)mm3, and the shape of 70% tumor was irregular, and some appeared as lobulated, and the tumor appeared as heterogeneous hypoechoic mass, no necrosis or liquefaction was found 28 days after implantation. There was low speed circular vasculature signal in around 70% tumors.Conclusions The orthotopic pancreatic cancer model in nude mouse can be established by in situ injection and this method is relatively ideal because it is simple and effective. The high frequency probe of endoscopic ultrasonograph is a reliable method for monitoring implanted pancreatic cancer.