Abstract: The loop-mediated isothermal amplification (LAMP) technique is a technique for the specific and efficient amplification of target fragments at a constant temperature using two pairs of specially designed primers and a strand displacement activity DNA polymerase. LAMP technique is a simple, rapid, specific, sensitive and cost-effective nucleic acid amplification method, and therefore has a promising future in the field rapid detection of Mycobacterium tuberculosis and grassroots applications. In this review, the basic principles and characteristics of the LAMP technique, the main molecular markers for the diagnosis of tuberculosis, and the use of different molecular markers and various types of novel techniques in the diagnosis of pulmonary tuberculosis, extrapulmonary tuberculosis, and drug-resistant tuberculosis were described. The LAMP technique has been widely used in the diagnosis of tuberculosis with high sensitivity and specificity, but the technique still has some shortcomings. This paper reviews the progress of its application in tuberculosis in recent years and provides an outlook on its development, with a view to providing a rational research direction for rapid diagnosis of tuberculosis in a resource-limited environment.

OBJECTIVETo evaluate the replication and encapsidation of HBV mutants with the truncated C gene.

METHODSThe HBV mutants with the truncated C gene were constructed by molecular cloning and PCR-based deletion in vitro. The replication and encapsidation of HBV mutants were investigated by Southern blotting, PCR and real-time fluorescence PCR respectively after transfecting the HBV mutants plasmid into HepG2 cells by using liposome.

RESULTSThe C-truncated HBV mutant vectors were constructed successfully and confirmed exactly by clone sequencing and enzymes digestion. The C-truncated HBV mutants were replication defective, however, all types of HBV DNA could be detected positive in the cytoplasm and supernatant after co-transfecting the C-truncated HBV mutants plasmid and the helper constructs into HepG2 cells. The C-truncated HBV mutants were proved to produce 3-40 folds more progeny DNA than that of the wild-type HBV by DNA quantitative assay.

CONCLUSIONThe C-truncated HBV mutants are replication-deficient and could not replicate and encapsulate in the hepatocytes when transfected solely, however, the progeny HBV-variant viruses are encapsidated more effectively to secrete into supernatant when co-transfected with the helper construct which lacks part of 5 prime-proximal HBV RNA packaging signal Epsilon.

Cell Line, Tumor ; Hepatitis B Core Antigens ; genetics ; Hepatitis B virus ; genetics ; physiology ; Humans ; Mutation ; Plasmids ; genetics ; Transfection ; Virus Replication

OBJECTIVETo review the development of tissue culture on Rehmannia glutinosa.

METHODDocumentaries at home and abroad were consulted.

RESULT AND CONCLUSIONThe tissue culture conditions of R. glutinosa were summarized and the problems of tissue culture in R. glutinosa were also pointed out.

Culture Media ; Lighting ; Plant Diseases ; Plant Leaves ; growth & development ; Plant Roots ; growth & development ; Plant Stems ; growth & development ; Plants, Medicinal ; growth & development ; Pollen ; growth & development ; Rehmannia ; growth & development ; Tissue Culture Techniques ; Tissue Preservation

OBJECTIVETo study the effects of C. versicolor petroleum ether extracts (CVPE) on the adenohypophysis androgen receptor level in mature castrated male rats.

METHODAll the rats in experiment were anesthetized for bilateral testicular and epididymis removal under sterile condition. The rats were randomized into four groups on the 14 th day after operation. The first group was intragastric physiological saline for castratered control group. The second group was intragastric CVPE 2 g x kg(-1) for low-dose group. The third group was high-dose group by giving CVPE 4 g x kg(-1). The fourth group was injected hypodermic testosterone propionate for positive-effect drug treatment group. The drug was given orally to animals one time a day successively for 21 days. The androgen receptor (AR) in adenohypophysis of mature castrated male rats was determined by the immunohistochemistry method and the level of serum testosterone (T) were determined by the radio-immunoassay after ig CVPE for 21 days.

RESULTThe immunohistochemistry results showed that positive cell numbers of androgen receptor in positive control and each CVPE groups were more than those in the castrated control group. The serum T level was increased greatly in mature castrated male rats treated with CVPE compared with the control group (P < 0.05, P < 0.01).

CONCLUSIONThe results show that CVPE can increase the adenohypophysis androgen receptor and serum T level in mature castrated male rats. It is indicated that CVPE has the effects on the hypophysis function.

Animals ; Lizards ; Male ; Materia Medica ; isolation & purification ; pharmacology ; Orchiectomy ; Pituitary Gland, Anterior ; drug effects ; metabolism ; Random Allocation ; Rats ; Rats, Wistar ; Receptors, Androgen ; metabolism ; Testosterone ; blood

OBJECTIVETo investigate the clinical effects of close reduction and proximal femoral locking plate fixation for the treatment of femoral neck fractures in young adults.

METHODSFrom August 2010 to 2014 August, 54 patients with displaced femoral neck fracture were treated with closed reduction and proximal cannulated screw locking plate fixation. There were 34 males and 20 females, aged from 18 to 55 years with an average of 39.8 years. The informations of fracture healing and complications were recorded after operation. According to Harris criteria, the function of hip joint was evaluated.

RESULTSAll patients were followed up from 4 to 24 months with an average of 11.3 months. Three cases occurred fracture nonunion, fracture union rate was 94.4%(51/54), union time was from 3 to 6 months with an average of 4.1 months. Among the healed 51 cases, the median of femoral neck shorten was 0.8 mm with the mean of (0.48±0.46) mm. No complications such as infection, internal fixation displacement were found during follow up. According to Harris criteria, 40 cases obtained excellent results, 9 good, 2 fair, 3 poor.

CONCLUSIONSClose reduction and proximal femoral locking plate fixation is an effective method in treating femoral neck fracture in young adults, it has advantages of avoiding the femoral neck crispation, reliable fixation, high rate of fracture union and good functional recovery.

OBJECTIVETo evaluate the clinical value of radial endorectal ultrasound (ERUS) in the assessment of preoperative staging of rectal carcinoma.

METHODSOne hundred and ten patients with rectal cancer underwent preoperative endorectal ultrasound (ERUS) examination in our hospital from February 2010 to September 2011. ERUS was performed using a Hitachi 900, Hitachi HI Vision Preirus US scanner, with a 5 - 10 MHz rigid rotating radial transducer and a focal length of 2 - 5 cm. The size, shape, echo pattern, infiltration depth, degree of circumferential involvement, extra-rectal invasion of the lesions and lymph node involvement were observed. The results of ERUS staging were compared with histopathological findings of the surgical specimens.

RESULTSThe accuracy of ERUS for T staging was 91.4%. The accuracy of ERUS in diagnosing stage T1, T2, T3, T4 cancers was 92.7%, 88.2%, 88.2% and 96.4%, respectively. The sensitivity of ERUS in diagnosing stage T1, T2, T3, T4 cancers was 92.3%, 72.7%, 85.4% and 71.4%, respectively. The specificity of ERUS in diagnosing stage T1, T2, T3, T4 cancer was 92.9%, 92.0%, 90.3% and 100.0%, respectively. Comparing the consistency of preoperative T-staging and postoperative pathological results, the Kappa value was 0.75, with a considerable consistency. The sensitivity, specificity, and accuracy of ERUS in the assessment of lymph node metastasis were 74.2%, 89.9% and 85.5%, respectively. Comparing the consistency of preoperative N-staging and postoperative pathological results, the Kappa value was 0.64, with a considerable consistency.

CONCLUSIONSERUS is a practical and accurate tool in assessment of preoperative staging of rectal tumors in regard to tumor invasion depth (T) and regional lymph node status (N), with advantages of simple operation, less pain, and high accuracy.

Adult ; Aged ; Endosonography ; methods ; Female ; Humans ; Lymphatic Metastasis ; Male ; Middle Aged ; Neoplasm Staging ; Preoperative Period ; Rectal Neoplasms ; diagnostic imaging ; pathology ; surgery ; Rectum ; diagnostic imaging ; pathology ; surgery

OBJECTIVETo explore the effect and mechanism on HBV replication in C gene truncated mutant.

METHODSProtein expression of C gene truncated vector and wild C gene vector were assay by SDS-PAGE Western blot. Constructed C gene truncated expression vector was cotransfected with wild HBV genome; virus load was detected by PCR in the culture medium and the cell. The formation of core particle was assay by Native western blot.

RESULTSThe recombinant vectors can efficiently express. Virus load of the cotransfected group by pcDNA3-deltaC and adwR9 was lower than that of control group in the culture medium and the cell. Protein band of the co-expressed group by pcDNA3-deltaC and pcDNA3-C showed slightly weaker than that of the co-expressed group by pcDNA3 and pcDNA3-C.

CONCLUSIONC gene truncated mutant could interfere with the formation of core particle and reduce of HBV replication

Cell Line ; Genetic Therapy ; Hepatitis B ; therapy ; Humans ; Mutation ; Transfection ; Viral Core Proteins ; genetics ; Virus Replication

Objective:To construct HIV-1 pseudovirus containing enhanced green fluorescent protein ( EGFP ) gene. To understand the interaction between the virus and the cells. Methods: HIV-1 pseudovirus containing EGFP gene was constructed by lentiviral packaging systems, and its EGFP gene was amplified using RT-PCR. The level of genomic integration and transcription of HIV-1 pseudovirus containing EGFP gene were detected on iDCs infected with HIV-1 pseudovirus. At the same time, research on expression of the EGFP gene in iDCs infected with HIV-1 pseudovirus was performed. Results:The EGFP gene of HIV-1 pseudovirus was detected through RT-PCR. The EGFP gene was identified in iDCs infected with HIV-1 pseudovirus through PCR and RT-PCR. The EGFP was observed in iDCs infected with HIV-1 pseudovirus under fluorescence microscopy. Conclusion: HIV-1 pseudovirus containing EGFP gene has been successfully produced. The HIV-1 pseudovirus that we constructed can infect iDCs,then its RNA can integrate into the genome of iDCs in the way of reverse transcription,and the EGFP gene could express in the iDCs after infected with HIV-1 pseudovirus.

Animals ; COS Cells ; Female ; Genetic Therapy ; Interleukin-12 ; genetics ; Liver Neoplasms, Experimental ; immunology ; therapy ; Mice ; Mice, Inbred BALB C ; T-Lymphocytes, Cytotoxic ; immunology

OBJECTIVETo evaluate the possibility of hepatitis B virus (HBV) as a vector in liver-targeting gene therapy.

METHODSA fragment containing the small envelope gene of HBV was replaced with the reporter gene green fluorescent protein (GFP) to construct the recombinant HBV vector, which was transfected into HepG2 cells with liposome. The expression of GFP was observed with fluorescence microscope. The HBV cccDNA was testified using semi-nest PCR. The viral particles of the recombinant HBV in culture medium were detected by PCR as well as Southern blot.

RESULTSThe HBV vector carrying the interesting gene of GFP could express the functional protein in the transfected hepatocytes. However, the recombinant HBV vector was replication-deficient, which could not be packed and replicated in the hepatocytes to secrete mature recombinant HBV particles carrying the interesting gene of GFP when transfected solely but could when cotransfected with the recombinant and helper construct which lacked part of 5'-proximal HBV RNA packaging signal epsilon.

CONCLUSIONIt is possible that HBV is reconstructed as a liver-targeting vector for gene therapy.

Cell Transformation, Viral ; Cells, Cultured ; Gene Transfer Techniques ; Genes, Reporter ; Genetic Therapy ; methods ; Genetic Vectors ; genetics ; physiology ; Hepatitis B virus ; genetics ; physiology ; Hepatocytes ; cytology ; virology ; Humans ; Liver ; cytology ; virology ; Recombinant Proteins ; genetics ; Transfection ; Virus Replication