Objective To establish a rapid and high sensitive assay of real-time fluorescence loop-mediated isothermal amplification assay for clinical detection of HIV-1 DNA.Methods Four loop primers were constructed for loop-mediated isothermal amplification ( LAMP) assay based on six conserved regions selected from HIV gene sequence .SYBR Green Ⅰdye was added into the established LAMP assay and its specificity and sensitivity were evaluated .Results The real-time fluorescence LAMP assay for the detection of HIV-1 DNA was successfully established .It was used for the detection of HIV-1 DNA among 200 patients with HIV infection, of which 195 cases were positive.Moreover, 50 healthy subjects were found HIV-1 DNA negative tested by the real-time fluorescence LAMP assay .Quantitative testing for HIV-1 DNA showed that the lowest and the highest detectable amount were 51 copies/ml and 8.21×106 copies/ml respectively, and the average amount was 5.78×105 copies/ml.HIV viral loads ranging from 1×105 to 10×105 were detected in 162 of 200 patients (83.08%).The ten times dilution method showed that the lowest detection limit of the assay was 50 copies/ml.The crossover experiment indicated that the specificity of the assay was 100%as none of HBV-DNA, HSV-DNA and HCV-RNA was determined by the assay .Conclusion The present study shows that 97.5%of the patients with HIV infection are confirmed HIV-1 DNA positive by the real-time fluorescence LAMP assay , suggesting that the real-time fluorescence LAMP assay is a rapid and sensi-tive assay with high specificity and could be applied for clinical detection of HIV-1 DNA.

Objective To evaluation the efficacy of dexmedetomidine mixed with dezocine and levobupivacaine for patient-controlled epidural analgesia (PCEA) after cesarean section.Methods A total of 300 patients, aged 23-35 yr, with body mass index of 24-28 kg/m2 , of American Society of Anesthesiologists physical status Ⅰ or Ⅱ , undergoing elective cesarean section under epidural anesthesia, were randomly divided into 3 groups (n =100 each) according to the random number table: morphine and levobupivacaine group (ML group), dezocine and levobupivacaine group (DL group), and dexmedetomidine, dezocine and levobupivacaine group (DDL group).In group ML, the loading dose included morphine 2 mg + levobupivacaine 10 mg + 5 ml normal saline, and PCEA solution contained morphine 5 mg + levobupivacaine 150 mg + normal saline 100 ml.In group DL, the loading dose included dezocine 3 mg+ levobupivacaine 10 mg+ normal saline 5 ml, and PCEA solution contained dezocine 15 mg + levobupivacaine 150 mg + normal saline 100 ml.In group DDL, the loading dose included dezocine 2 mg+ levobupivacaine 10 mg + dexmedetomidine 0.5 μg/kg + normal saline 5 ml, and PCEA solution contained dezocine 7.5 mg+ levobupivacaine 150 mg + dexmedetomidine 1.5 μ g/kg + normal saline 100 ml.At 10 min before the end of operation, the loading dose was given via the epidural catheter, and the PCEA pump was connected and set up to deliver a 2 ml bolus dose with a 15-min lockout interval and background infusion at 2 ml/h.Analgesia was maintained until 42 h after operation.Visual analog scale score was main-tained ≤ 3, and (or) visual analog scale for fatigue score ≤ 2.When PCEA failed, morphine 2 mg was injected epidurally as rescue analgesic.At 4, 8, 24 and 42 h after operation, the modified Bromage score and Ramsay sedation score were recorded, and patients' satisfaction with analgesia was evaluated.The occurrence of epidural analgesia-related adverse reactions was recorded.Results The consumption of PCEA solution and requirement for rescue analgesics were significantly lower in group ML than in group DL.Compared with group ML, Ramsay sedation score was significantly decreased at 4 and 8 h after operation, the degree of patients' satisfaction with analgesia was increased, and the incidence of nausea, vomiting, dizziness and pruritus was decreased in DL and DDL groups, and the incidence of urinary retention was decreased in group DDL.The degree of patients' satisfaction with analgesia was significantly higher, and the incidence of dizziness and urinary retention was lower in DDL group than in group DL.No patients developed epidural analgesia-relatcd hypotension, bradycardia, or respiratory depression in the three groups.Conclusion The efficacy of dexmedetomidine mixed with dezocine and levobupivacaine is good when used for PCEA after cesarean section, and the adverse reactions are fewer.

OBJECTIVETo detect miR-143 expression in human nasopharyngeal carcinoma (NPC) cell lines and explore the role of miR-143 in regulating the adhesion ability of NPC cells.

METHODSFluorescence quantitative RT-PCR was used to detect miR-143 expression levels in 5 NPC cell lines (CEN1, CNE2, HONE1, 5-8F, and 6-10B) and an immortalized human nasopharyngeal epithelial cell line (NP69). The retrovirus plasmid pMSCV-puro-miR-143 was constructed and the packaged retroviruses pMSCV-puro and pMSCV-puro-miR-143 were infected in 5-8F cells to establish a cell line with stable miR-143 overexpression, whose adhesion ability was tested with adhesion assay.

RESULTSThe expression of miR-143 was down- regulated in the NPC cell lines, and the highly metastatic NPC cell line 5-8F had a expression of only 3.0% of the control level, as compared with the level of 63.59% in the tumorigenic but nonmetastatic NPC cell line 6-10B. The transfected 5-8F cells showed a 1080-fold increase of miR-143 expression (P<0.05) with a significantly lowered adhesion ability.

CONCLUSIONmiR-143 plays a role in regulating the invasiveness and metastasis of NPC, and overexpression of miR-143 causes a significant reduction of the adhesion ability of the highly metastatic NPC cell line 5-8F.

Carcinoma ; Cell Adhesion ; Cell Line, Tumor ; Cell Movement ; Humans ; MicroRNAs ; metabolism ; Nasopharyngeal Neoplasms ; metabolism ; pathology

Objective:To preliminarily observe the clinical efficacy of microwave hyperthermia combined with intensity-modulated radiotherapy (IMRT) and chemotherapy for patients with locally advanced gastric cancer.Methods:Forty patients who could not been operated or refused operation were enrolled in this clinical trial, who were confirmed as locally advanced proximal or distal gastric cancer by gastroscopy pathology and imaging. Radiotherapy was delivered by IMRT technology for 5 times per week with a total dose of 46 to 56 Gy (median dose of 50 Gy) in 25 to 28 fractions. Synchronous hyperthermia was given at 42 to 44℃ twice a week, 45 min/time. S-1 or capecitabine-based synchronous chemotherapy was performed, d1-14/3 weeks. The symptom remission rate, adverse reactions, objective remission rate (complete and partial remission) and survival were observed.Results:A total of 40 patients, aged between 56 and 83 years (median age of 71 years), were enrolled in this study. The male-to-female ratio was 7: 1. Among them, 38 cases (95%) showed symptom remission. The most common adverse reactions were grade 1-2 gastrointestinal reactions and leukopenia. The objective remission rate was 87.5%, the 2-year progression-free survival and overall survival rates were 68.6% and 70.5%, respectively.Conclusion:Preliminary findings demonstrate that microwave hyperthermia combined with chemoradiotherapy achieve satisfactory outcomes and yield tolerable toxicity in patients with locally advanced gastric cancer.