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 discuss the application principle in tuina manipulation for lumbar intervertebral disc herniation (LIDH) in Chinese literatures published in recent 30 years. Methods: The three major Chinese databases, Wanfang Academic Journal Full-text Database (Wanfang), Chongqing VIP Database (CQVIP) and China National Knowledge Infrastructure (CNKI), were searched to collect the studies of tuina manipulations in treatment of LIDH published in recent 30 years. Clustering analysis was applied to analyze the top 20 tuina manipulations for LIDH. Results: The top 20 most frequently used manipulations for LIDH were Gun-rolling, Rou-kneading, Dian-digital pressing, oblique Ban-pulling, An-pressing, Tanbo-plucking, Bashen-pulling and extending, horizontal Tui-pushing, Na-grasping, Anrou-pressing and kneading, Dou-shaking, Yao-rocking, Ca-scrubbing, Pai-patting, post-extension Ban-pulling, Mo-rubbing, Zhen-vibrating, Nie-pinching, fist-back Ji-tapping, and dorsal Shen-extending methods. The involved manipulations can be divided into two categories by the treated body areas. One category is applied to the soft tissues, including Gun-rolling, Rou-kneading, Dian-digital pressing, An-pressing, Tanbo-plucking, horizontal Tui-pushing, Na-grasping, Anrou-pressing and kneading, Ca-scrubbing, Pai-patting, Mo-rubbing, Zhen-vibrating, Nie-pinching, and fist-back Ji-tapping methods. The other category is applied to bones and joints, including oblique Ban-pulling, Bashen-pulling and extending, Dou-shaking, Yao-rocking, post-extension Ban-pulling, and dorsal Shen-extending methods. Conclusion: Based on the treated body area, the tuina manipulations applied to treat LIDH are predominated by the ones performed on soft tissues, assisted by those on bones and joints. From the way of force exertion, the involved manipulations are majorly the swinging methods, followed by squeezing and pressing ones. The manipulations applied to bones and joints are predominated by the Ban-pulling ones, followed by the Bashen-pulling and extending ones.

Objective To determine the distribution of HCV genotypes in patients with chronic hepatitis C,study the distribution of genotype in different gender and the relationship between genotypes and serum HCV-RNA levels.Methods Two hundred and six cases of HCV RNA positive patients(all with relevant clinical data) receiving pegylated interferon therapy were collected from May to December 2010.HCV RNA was detected in 206 hepatitis C patients from 40 hospitals in China by Roche Cobas AmpliPrep/Cobas TaqMan HBV test,and genotype was determined by Abbott RealTime HCV G enotype Ⅱ .The distribution of genotypes in the gender was analyzed by chi-square test analysis.The relationship between genotypes and serum HCV RNA levels was detected by single factor analysis and two independent sample t test analysis.Results There were seven different subtypes of HCV in 206 samples,including genotype 1,7 cases(3.4% ,7/206); genotype 1a,2 cases(1.0%,2/206); genotype 1b,123 cases (59.7 %,123/206); genotype 2,32 cases(15.5 %,32/206); genotype 3,27 cases(13.1%,27/206); genotype 6,6 cases(2.9% ,6/206) ;genotype 1/6,5 cases(2.4% ,5/206) ;genotype 2/4,1 cases(0.5%,1/206).There was no significant difference between HCV genotype and gender in 132 cases with genotype 1 and 65 cases with non-genotype 1(genotype 2,3,6) (x2 = 0.000,P ＞ 0.05).There was significant association between quantity of HCV RNA and genotype in 188 patients with HCV(F = 3.371,P＜ 0.01).The 197 patients with HCV single genotype were divided into five groups in terms of region(East,South,West,North and Center).There was no significant difference between HCV genotype 1 and non-genotype 1 in the five groups(x2 = 5.840,P ＞ 0.05).Conclusions It is suggested that HCV 1 b is the most prevalent type in China,followed by HCV 2.There is no significant difference between HCV genotype and gender.The levels of HCV RNA with genotype 1b are significantly higher than those with genotype 3.The levels of HCV RNA with genotype 2 are significantly higher than those with genotype 3.The levels of HCV RNA with genotype 6 are significantly higher than those with genotype 3.

Aged ; Balloon Valvuloplasty ; methods ; Coronary Artery Bypass, Off-Pump ; methods ; Coronary Artery Disease ; surgery ; Humans ; Male ; Middle Aged ; Pulmonary Valve Stenosis ; surgery

In this study, to investigate the effect on expression of IL-2 in lymphocytes from bone marrow and peripheral blood of normal donors after they were mobilized by G-CSF in allo-BMT, 7 normal donors bone marrow and peripheral blood were harvested before and after G-CSF administration. The separated lymphocytes were measured by FCM after they were stained intracellularly by anti-IL-2, and their expressions of IL-2 were compared. The degree of aGVHD in patients after bone marrow transplantation was evaluated clinically, and it was compared with the status of aGVHD of 15 patients whose donors didn't receive G-CSF administration in our department, and 2 groups of patients are comparable in age, types of diseases and status of donors. The results showed that the expression of IL-2 in lymphocytes in 7 G-CSF mobilized donors decreased significantly after G-CSF administration and more severe aGVHD than grade II didn't develop in these recipient patients, and comparing with 15 patients received the bone marrow from donors who didn't receive G-CSF, the incidence of aGVHD decreased. It is suggested that the expression of IL-2 in lymphocytes was influenced by donors' G-CSF administration, and it is likely that thereby reduces the incidence of aGVHD in patients after BMT.
Adult ; Blood Donors ; Bone Marrow Cells ; drug effects ; metabolism ; Bone Marrow Transplantation ; adverse effects ; Female ; Flow Cytometry ; Graft vs Host Disease ; etiology ; Granulocyte Colony-Stimulating Factor ; pharmacology ; Hematopoietic Stem Cell Mobilization ; Humans ; Interleukin-2 ; biosynthesis ; Lymphocytes ; drug effects ; metabolism ; Male

OBJECTIVETo better understand the clinical feature of viral gastroenteritis attributed to noroviruses and to summarize the experience on an outbreak of acute gastroenteritis through rapidly colleting and confirmation of related information regarding to noroviruses in hospitals.

METHODSInformation on an outbreaks involving 18 patients with acute gastroenteritis in one hospital regarding its epidemiological and clinical features and to perform bacteria culture for stool specimens on every patient. On 7 patients, rotavirus antigen were RNA tested together with norovirus nucleic acid were examined by ELISA and PAGE and RT-PCR.

RESULTS(1) Most of the patients were elderly with several chronic diseases. (2) Watery diarrhea (12/18, 66.67%) and few with mucous (3/18, 16.67%) were seen. Most stool examination was normal (10/18, 55.56%) but few stool specimen could be found with some leucocytes (3/18, 16.67%) and little occult blood (4/18, 22.22%). (3) All bacteria culture in stools showed negative. There was no rotavirus RNA identified but 3 specimen showed norovirus nucleic acid positive as 42.86% (3/7).

CONCLUSIONNorovirus was one of the important pathogens causing acute gastroenteritis outbreaks in hospitals attacking elderly with several chronic diseases in particular. Surveillance program targeting elderly inpatient with diarrhea should be enhanced, especially in autumn and winter.

Acute Disease ; Aged ; Caliciviridae Infections ; epidemiology ; China ; epidemiology ; Disease Outbreaks ; Enzyme-Linked Immunosorbent Assay ; Feces ; virology ; Gastroenteritis ; epidemiology ; virology ; Hospitals ; statistics & numerical data ; Humans ; Norovirus ; isolation & purification ; Reverse Transcriptase Polymerase Chain Reaction

OBJECTIVE(1) To investigate the expression and gene diversity of the 7 major cancer/testis (CT) antigens, MAGE-1, MAGE-3, MAGE-4, MAGE-10, NY-ESO-1, SSX-2 and SCP-1, in hepatocellular carcinoma (HCC). (2) To analyze the correlations between the clinical characters and CT antigens' expression.

METHODSThe cancer and para-cancer tissues were collected from 30 HCC patients. The mRNAs of seven CT antigens were detected by reverse transcription-polymerase chain reaction (RT-PCR) with the specific primers. The PCR products were sequenced to analyze the CT genes.

RESULTSThe MAGE-1, MAGE-3, MAGE-4, MAGE-10, NY-ESO-1, SSX-2 and SCP-1 were expressed in 66.7%, 70.0%, 20.0%, 36.7%, 40.0%, 33.3% and 33.3% of the tumor tissues from HCC patients respectively, however, they were not expressed in the para-cancer tissues. Among the 30 patients investigated, 90.0% expressed one CT gene at least, 70.0% expressed two CT genes, and 53.3% expressed three CT genes of the seven CT genes. The coding genes of these CT antigens were highly conserved between in Chinese patients and patients abroad. There were discernible correlations between alpha-fetoprotein level and MAGE-10 or SCP-1 expression level, as well as between average age and MAGE-3 or SSX-2 expression levels (P<0.05).

CONCLUSIONSWith a highly conserved coding gene, seven CT antigens were expressed in 20.0% - 70.0% of Chinese HCC patients. CT antigens' expression had correlations with some clinical characters.

Antigens, Neoplasm ; biosynthesis ; genetics ; Carcinoma, Hepatocellular ; genetics ; immunology ; Female ; Gene Expression Regulation, Neoplastic ; genetics ; Humans ; Liver Neoplasms ; genetics ; immunology ; Male ; Melanoma-Specific Antigens ; Neoplasm Proteins ; biosynthesis ; genetics ; RNA, Messenger ; biosynthesis ; genetics

OBJECTIVES(1) To evaluate the prevalence, phenotypes and suppressive function of CD4+CD25+ regulatory T cells (Tregs) among the in peripheral blood mononuclear cells (PBMCs) and tumor-infiltration lymphocytes (TILs) from hepatocellular carcinoma (HCC) patients and patients with chronic hepatitis B. (2) To investigate the correlation between the frequency of CD4+CD25+ Tregs and clinical characteristics of HCC patients.

METHODSPBMCs and TILs in 18 HCC patients, 10 chronic hepatitis B (CHB) patients and 15 healthy donors were evaluated for the phenotypes of CD4+CD25+ Tregs and the proportion of CD4+CD25+ Tregs as a percentage of the total CD4+ cells, by flow cytometric analysis with three or four color staining. The relationship between the frequency of CD4+CD25+ Tregs and tumor TNM stages was analyzed. The CD4+CD25+ Tregs and CD4+CD25- T cells were isolated from PBMC of HCC patients and donors. The suppressive function of CD4+CD25+ Tregs was analyzed.

RESULTSThe percentages of CD4+CD25+ Tregs of the HCC patients (6.38% +/- 6.30%) and CHB patients (4.29% +/- 1.82%) were significantly higher than those of the healthy donors (1.58% +/- 0.55%, P less than 0.01). Among the TILs, the percentage of CD4+CD25+ Tregs was higher (t = 4.39, P < 0.01). There were significant differences in the prevalence of CD4+CD25+ Tregs in early and advanced stage HCCs (stage II vs. III, P less than 0.05; stage II vs. IV P < 0.01). The proliferative capacity of CD4+CD25- T cells was inhibited by the presence of CD4+CD25+ T cells in a dose-dependent manner where the level of suppression was correlated to the ratio of the two-cell populations.

CONCLUSIONThese results suggest that the increase in frequency of CD4+CD25+ Tregs might play a role in the suppression of the immune response against HCC, which may contribute to the HCC cells that escaped from immunological surveillance.

Adult ; Aged ; Carcinoma, Hepatocellular ; metabolism ; Female ; Humans ; Interleukin-2 Receptor alpha Subunit ; Liver Neoplasms ; metabolism ; Male ; Middle Aged ; T-Lymphocytes, Regulatory ; immunology ; Young Adult

OBJECTIVETo explore the mechanism of tanshinol on alleviate the inflammatory injury of lung tissue in rat hepatopulmonary syndrome (HPS).

METHODSSD rats were randomly divided into normal control group (n = 8), hepatopulmonary syndrome (HPS) group (n = 11) and tanshinol intervention group (n = 9). HE staining was used to observe the histopathology changes of pulmonary and hepatic tissues, and to count the number of macrophages in lung tissues. The activity of alanine transferase (ALT) and concentrations of endotoxin, tumor necrosis factor-a (TNF-alpha) and homocystein (Hcy) in plasma were detected. The concentrations of TNF-alpha, nitric oxide (NO) and malondialdehyde (MDA) and the activity of inducible nitric oxide synthase (iNOS) in the lung tissues were measured, respectively.

RESULTSThickened alveolar septum and increased macrophages were observed in lungs in HPS rat. After administered with tanshinol, the pulmonary pathological changes were alleviated and the number of macrophages in lung tissue was decreased compared with HPS group. The activity of ALT and the concentrations of endotoxin, TNF-alpha and Hcy in plasma ,and TNF-alpha, iNOS, NO and MDA in lung tissue in HPS group were higher than those of normal control group; meanwhile, those tanshinol group were less those that of HPS group.

CONCLUSIONTanshinol may play an important role in delaying the development of HPS through protecting liver or directly antagonizing the effect of intestinal endotoxemia so as to alleviate the inflammatory reaction in lung tissue.

Alanine Transaminase ; metabolism ; Animals ; Caffeic Acids ; pharmacology ; Disease Models, Animal ; Endotoxins ; blood ; Hepatopulmonary Syndrome ; drug therapy ; pathology ; Homocysteine ; blood ; Liver ; drug effects ; pathology ; Lung ; drug effects ; pathology ; Macrophages ; drug effects ; pathology ; Male ; Malondialdehyde ; metabolism ; Nitric Oxide ; metabolism ; Nitric Oxide Synthase Type II ; metabolism ; Rats ; Rats, Sprague-Dawley ; Tumor Necrosis Factor-alpha ; blood

OBJECTIVETo identify the types of voltage-dependent Ca2+ channels (VDCCs) in human ejaculated sperm using molecular technology.

METHODSBased on the WHO criteria, the donated semen was screened by computer-aided sperm analysis (CASA), the sperm optimized by swim-up method, and the VDCCs' alpha1 subunits examined by reverse transcription polymerase chain reaction (RT-PCR).

RESULTSSuch mRNA messages as alpha1H, alpha1G, alpha1E, alpha1B and alpha1C were detected, but alpha1A and alpha1D were not.

CONCLUSIONHuman motile sperms express multiple VDCC RNAs, among which T-type and non-L-type channel messages are likely to be predominantly expressed. Not only T-type but also non-L-type calcium channels may be major gates for the external calcium influx, which helps regulate sperm motility and acrosome reaction.

Calcium ; metabolism ; Calcium Channels ; biosynthesis ; classification ; genetics ; Humans ; Image Processing, Computer-Assisted ; Male ; RNA, Messenger ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Spermatozoa ; metabolism