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.

Objective To explore the establishment of a risk prediction model for concurrent bronchiolitis obliterans(BO)in children with refractory Mycoplasma pneumoniae pneumonia(RMPP).Methods A retrospective study included 116 RMPP children treated in the Department of Pediatrics of Xiangya Changde Hospital from June 2021 to December 2023.Eighty-one cases were allocated to the training set and thirty-five cases to the validation set based on a 7:3 ratio.Among them,26 cases in the training set developed BO,while 55 did not.The multivariate logistic regression was used to select variable factors for constructing the BO risk prediction model.Nomograms were drawn,and the receiver operating characteristic(ROC)curve was used to assess the discriminative ability of the model,while calibration curves and Hosmer-Lemeshow tests evaluated the model's calibration.Results Multivariate logistic regression analysis indicated that several factors were significantly associated with concurrent BO in RMPP children,including length of hospital stay,duration of fever,atelectasis,neutrophil percentage(NEUT%),peak lactate dehydrogenase(LDH),ferritin,peak C reactive protein(CRP),oxygenation index(PaO2/FiO2),≥2/3 lung lobe consolidation,pleural effusion,bronchial mucous plugs,bronchial mucosal necrosis,and arterial oxygen partial pressure(PaO2)(P<0.05).ROC curve analysis for the training set indicated an area under the curve of 0.904 with 88%sensitivity and 83%specificity;the validation set showed an area under the curve of 0.823 with 76%sensitivity and 93%specificity.The Hosmer-Lemeshow test's Chi-square values for the training and validation sets were 2.17 and 1.92,respectively,with P values of 0.221 and 0.196,respectively.Conclusions The risk prediction model for BO in RMPP children based on logistic regression has good performance.Variables such as length of hospital stay,duration of fever,atelectasis,peak LDH,peak CRP,NEUT%,ferritin,≥2/3 lung lobe consolidation,pleural effusion,bronchial mucous plugs,bronchial mucosal necrosis,PaO2/FiO2,and PaO2 can be used as predictors.

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 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 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 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.

Objective To investigate the effects of N -acetylcystein ( NAC) on the cardiac hypertrophy in pressure -overload rats and the relationship with the expression of inflammatory cytokines.Methods The SD rats were randomly divided into four groups:sham group, model group , drug group in sham and drug group in model.Pressure-overload rat model was established by abdominal aortic coarctation.Rats in drug group were treated with NAC ( 0.2 g? kg-1? d -1 , ig ) for 8 weeks. Cardiac hypertrophy was evaluated by heart histological analysis and echocardiography.The collagen content was determined by chromatome-try.RT-PCR and ELISA were used to determine the expression of in-flammatory cytokines.The production of reactive oxygen species ( ROS) were measured by Fenton reaction.Results Compared with sham rats, blood pressure, left ventricular weight index( LVWI) , myocyte diameter, left ventricular wall thick(LVWT) as well as the collagen content( CC) in model rats significantly elevated ( P <0.05 ) .In model rats treated with NAC, LVWI, LVWT and CC were significantly decreased as well as the production of ROS ( P <0.05 ) . Furthermore, the expression of interleukin-6(IL-6)and tumor necrosis factor-α(TNF-α)but IL-10 was elevated in myocardium of pressure-o-verload rats, which was inhibited by NAC( P<0.05).NAC had no effects on the expression of IL-10.Conclusion NAC could improve the cardiac hypertrophy induced by pressure-overload.The effects of NAC probably relates to the decrease of pro-inflammatory cytokines expression in rat myocardium.

Non-neonatal tetanus is an acute, specific, toxic disease in patients over 28 days of age, characterized by continuous rigidity and paroxysmal spasms of the skeletal muscles throughout the body caused by the intrusion of Clostridium tetani through skin or mucosal membrane into the body and reproducing in anaerobic environments to produce exotoxins. The mortality rate of severe patients is close to 100% without medical intervention. Even with aggressive comprehensive treatment, the global mortality rate remains at 30%-50%, making it a potentially fatal disease. In order to standardize the diagnosis, treatment and prevention of non-neonatal tetanus, based on "Regulation for Diagnosis and Treatment of Non-neonatal Tetanus (2019 Edition)", experts have revised this regulation according to clinical practice and recent research progress in this field to guide medical institutions in the prevention and control of non-neonatal tetanus. This article interprets the key points and basis for updating the 2024 edition regulation to guide clinical implementation and application.

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