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

Aged ; Angioplasty, Balloon, Coronary ; adverse effects ; methods ; Female ; Hematoma ; etiology ; Humans ; Myocardial Infarction ; therapy ; Peritoneal Diseases ; etiology ; Radial Artery

Objective To investigate the clinical efficacy,operative essentials and indications of hybrid spinal fusion surgery for cervical spondylotic myelopathy.Methods From August 2008 to December 2011,thirty-eight patients with cervical spondylotic myelopathy underwent hybrid spinal fusion surgery in our hospital.There were 27 males and 11 females,aged from 33 to 70 years (average,51 years).A total of 86 segments were treated (fusion 48 vs.non-fusion 38).Twenty-eight patients underwent a two-level surgery,and ten patients received a three-level surgery.The Japanese Orthopaedic Association (JOA) score and Visual analogue scale (VAS) were used to evaluate pre-and post-operative neurological function and pain,respectively.The pre-and post-operative range of motion of the cervical spine was measured according to Xrays.Moreover,the surgical complications were recorded and analyzed.Results Thirty-seven patients were followed up for 15 to 55 months (average,29.1 months).The improvement of neurological function was obtained in 36 patients.The JOA score was improved from preoperative 10.5±1.57 to 14.3±1.97 at final follow-up,with an improvement rate of 58.46％,and the results were excellent in 16 cases,fair in 20 cases and poor in 1 case.The VAS was improved from preoperative 7.3±1.04 to 3.2±1.41 at final follow-up.The Cobb angle changed from preoperative 25°±3.21°to 20°±2.56°at final follow-up.After operation,the neurological function was not restored in 1 case; hoarseness and bucking occurred in 2 cases; sore throat occurred in 22 cases; anterior displacement of prosthesis (PCM) occurred in 3 cases.Other patients had no complications,such as displacement,loosening and heterotopic ossification.Conclusion In hybrid spinal fusion surgery,the lesions segments are decompressed fully,the severely degenerative segments are fused,and the motion of the non-fusion segments is reserved.As a result,not only the stability of the cervical spine is achieved,but also an obvious improvement of symptoms and a satisfactory short-term efficacy can be obtained.Therefore,this method is an alternative procedure for cervical spondylosis myelopathy.

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.

Objective To explore the influnce on the endothelin-1 (ET-1) and clinical application value of continuous blood purification (CBP) on the treatment of severe craniocerebral injury with the neurogenic pulmonary edema (NPE).Methods All data about sixty patients with NPE were prospectively studied.These 60 patients were randomly (random number) divided into control group (n =30) and treatment group (n =30).In control group,patients were rapidly given with lowering intracranial pressure,mechanical ventilation,calming,antibiotic therapy and so on.In the treatment group,patients received CBP integrated with routine treatment.On admission and 72 h posttreatment,ET-1,static lung compliance and oxygenation index were observed.Time of mechanical ventilation support,incidence rates of multipal organ dysfunction syndrome (MODS) were compared between two groups.The paired t-test was used for the amount data within the group.Chi-square was used for the constitute ratio and incidence ratio of the each relevant information.P ＜ 0.05 was considered statistically significant.Results Compared to the control group,the level of ET-1 was decreased significantly in the treatment group [(48 ± 10) ng/L vs.(85 ± 14) ng/L] after 72 h post-treatment,while static lung compliance [(60.9 ± 2.3) mL/cmH2O vs.(31.4 ±4.8) mL/cmH2O] and oxygenation index [(317 ± 11) mmHg vs.(192 ± 14) mmHg] increased significantly (P ＜ 0.05).In treatment group and control group,the time of respirator intervention were [(6.0 ± 2.1) d vs.(11 ± 3.2) d],and the statistical significance was shown (P ＜ 0.05).Compared to the control group [56.7％ (17/30)],incidence rate of MODS [20.0％ (6/30)] was lower in treatment group (P ＜ 0.05).Conclusions CBP combined with routine treatment,which can remove ET-1 effectively,improve oxygenation,reduce the time of mechanical ventilation support and incidence rate of MODS.

[Objective] To investigate combined application of multiplex reverse transcription-polymerase chain reaction(mRT-PCR)and karyotype analysis detect of clonal chromosomal aberrations in acute lymphoblasfic leukemia (ALL),and explore the expression of common fusion genes.Methods 189 ALL patients were examined by multiplex RT-PCR and R or G banding techniques.[Results]10 fusion genes were detected in 69 out of 189 ALL patients(36.5％),including E2A/PBX1,TEL/AML1,BCR/ABL,MLL/AF4,MLL/AF6、MLL/AF9,MLL/AF10,MLL/ELL,SIL/TAL1,TLS/ERG.R or G banding techniques could find chromosome structural and numeracy abnormalities in 86 out of 152 patients (56.6％) available for analysis.Combination of mRT-PCR and R or G banding could raise the rate of detecting clonal chromosomal abnormalities to 69.3％.Fusion genes were detected in 33 out of 90 (36.7％) patients with adult ALL and 36out of 99 (36.4 ％) patients with children ALL,there were 22 patients with positive BCR/ABL but no TEL/AML1 in adult ALL group,while there were 24 patients with positive TEL/AML.1 and 2 with positive BCR/ABL in children ALL group.There was significant statistical difference for the expression of RCR/ABL and TEL/AML1 between adult ALL and children ALL (P＜0.01),but no difference for MLL related fusion gene,E2A/PBX1,SIL/TAL1 and TLS/ERG(P＞0.05).BCR/ABL and TEL/AML1 fusion gene could be detected in 66 ALL patients with normal karyotype (36.3％).[Conclusion]There were different biological characteristics between adults and children with ALL.mRT-PCR technique can quickly screen chromosome structural aberrations in patients with newly diagnosed leukemia.It is useful in detection of fusion genes in ALL with normal karyotypes and it would refine the karyotype analysis and provide imrootramt prognosis-relevant information.

Objective To explore the value of fluorescence in situ hybridization (FISH) and multiplex RT-PCR in the detection of mixed lineage leukemia (MLL) gene rearrangement in acute leukemia (AL) patients. Methods Dual-color MLL probe, multiplex RT-PCR and R or G banding techniques were used to detect the MLL gene rearrangement in 189 cases of AL.Results MLL gene rearrangements were detected in 9 cases (5.03 ％) by FISH,and 16 cases (8.47 ％) by multiplex RT-PCR,including MLL/AF9,MLL/AF10,MLL/AF6, MLL/AF7, MLL/ELL, MLL/PTD. R or G banding techniques could find 11q23 in 5 out of 189 patients (2.65 ％). There was no statistic difference in the incidence of 6 common MLL gene rearrangements between ALL (73 cases) and AML patients (116 cases) (P ＞ 0.05).Conclusion Multiplex RT-PCR is a powerful technique in the detection of MLL gene rearrangement for tentatively diagnosed AL.It could not only confirm translocation detected by conventional cytogenetic method, but also detect MLL partial tandem duplication which could not been detected by cytogenetic examination or FISH. It plays an important role in guiding therapy and predicting prognosis for AL.

OBJECTIVETo establish sleeve gastrectomy(SG) rats model of obese type 2 diabetes mellitus(T2DM) for the research of hypoglycemic mechanism.

METHODSNine male Sprague-Dawley (6-week-old) rats were fed with high-sucrose and high-fat diet for 4 weeks, developing diet-induced obesity (DIO) rats model. The rats were then randomly divided into two groups. Six rats of them underwent sleeve gastrectomy(SG) as the sleeve gastrectomy group[SGG, body weight (471.8±17.9) g] and the other three rats underwent a laparotomy and stomach manipulation as the sham operative group[SOG, body weight (467.0±8.4) g]. The body weight, caloric intake and peripheral blood concentration of total ghrelin of rats were recorded after operation.

RESULTSThe weight of all the rats declined progressively after operation. The weight of the rats in SOG began to rise on the 5th postoperative day(POD) and regain their preoperative levels on the average 22nd POD. However, the weight of the rats in SGG began to rise slowly from the 9th POD, but was still lower than that of SOG[(487.4±10.1) g] and preoperative levels[(471.8±17.9) g] on the 28th POD[(420.1±18.6) g](P=0.001). Average caloric intake of rats in SGG was significantly lower than that of SOG after operation, but there was no significant difference between the two groups(P=0.121). The ghrelin level of SGG showed a continuous decreasing trend after intervention, decreased by 17.4% compared with the preoperative level (1595.1±14.4 ng/L) on the 28th POD[(1316.8±14.8) ng/L]. The ghrelin level of SOG did not change obviously before and after operation and both groups differ statistically(P=0.000).

CONCLUSIONSA SG rat model is successfully established. This model can be used for the further study of mechanism analysis of T2DM resolution after surgery.

Animals ; Diabetes Mellitus, Type 2 ; pathology ; Disease Models, Animal ; Gastrectomy ; methods ; Ghrelin ; blood ; Male ; Obesity ; blood ; surgery ; Rats ; Rats, Sprague-Dawley

Objective To elucidate the CT characteristics and pathology of acute miliary pulmonary tuberculosis (AMPT). Methods The CT features of AMPT in 25 cases were analyzed retrospectively, and the CT features in HIV-seronegative and HIV-seropositive patients were compared by 2-sided exact propability Chi-square test. Two lung specimens were inflated and fixed by Heitzman's method. HRCT scans, gross specimen section (80-150 μm) and histologic section (5 μm) were performed on dry lung specimens and CT-pathologic correlation was conducted. The distribution of micronodules in the secondary lobule on HRCT and pathology in one specimen was evaluated by Chi-square test. Results Twenty five patients with AMPT were included in this study, including 11 HIV-seropositive patients and 14 HIV-seronegative patients. HRCT showed diffuse micronodules randomly distributed throughout both lungs in 25 patients, and ground-glass opacity (17 patients)was the predominant complicated finding. Coalescence of nodules and consolidation in HIV-seropositive patients (5 and 6 patients) were markedly higher than that in HIV-seronegative patients (none). In lung specimens, most nodules located in the lung parenchyma between the central bronchovascular bundle and the perilobular structures (792 and 560 nodules), which located in the interlobular septum pathologically. The distribution of micronodules in the secondary lobule showed on HRCT (1060 nodules)and pathology(864 nodules) was not significantly difference(x2=2.814,P＞0.05). HRCT showed ground-glass opacities when ARDS occured, which were pulmonary edema,inflammation and hyaline membrane on alveolar wall pathologically. Conclusions The HRCT characteristic of nodule distribution in AMPT is random. ARDS should be suspected when diffuse ground-glass opacities appear on HRCT.

Objective To investigate the feature,pathological diagnosis and differential diagnosis of blastic plasmacytoid dendritic cell neoplasm (BPDCN).Methods The clinical and histological findings,immunophenotype,treatment and prognosis of one case with BPDCN were evaluated with review of the relevant literature.Results The female patient with age of 32 years presented as cutaneous lesions.Pathologic findings showed skin and bone marrow were involved.Microscopically,the tumors were composed of diffuse blasts with small to medium size,scant cytoplasm,highly irregular nuclear contours,dispersed nuclear chromatin and faint indistinct nucleolus.Immunohistochemical characteristics of the tumors were the positive expression of CD4,CD56 and CD123,and negative expression of CD3,CD20,CD34,TdT,MPO.Conclusion BPDCN is a rare and highly aggressive hematopoietic neoplasm.Except for the skin,bone marrow and lymph node can also be involved.The differential diagnosis is necessary from other haematopoietic and lymphoid tumors.