Abstract: Objective　To analyze the emergency response and long-term intervention effects after the detection of infectious snails epidemic by loop-mediated isothermal amplification (LAMP) assays in Hannan District, Wuhan City, and to explore the application of LAMP in early surveillance and early-warning of schistosomiasis transmission. Methods Snails picked up by the risk monitoring system in Hannan District were examined by anatomical microscopy and LAMP technology to identify the schistosomiasis infection. Emergency response and intensive intervention were initiated in the environment where positive snails appeared, and the long-term effects were evaluated. Results In May 2018, the infectious snails were detected by LAMP technology in Hannan District, and the positive snails were located in Zhujiacha, Dongzhuang Village, Obstacles and weeds were removed and buried by machine in Zhujiacha. 12 700 m2 of snails were killed by drugs, and the mortality rate of snails was more than 80%; no new seropositive persons were found in the emergency examination within 500 m of the positive snail sites. 506 people were examined in Dong Zhuang Village at the end of the year, and 30 positive IHA cases were detected with a blood positive rate of 5.93%, no positive fecal test was found, and all positive blood test patients took preventive medication. The monitoring results of sentinel rats and wild feces were all negative. Health education was carried out, 7 warning signs were deployed and refreshed, and 500 publicity brochures were distributed. After nearly three years of intensified intervention and monitoring in the villages where the positive environment is located, and the density of snails on the stubborn snail has dropped from 0.094/frame to 0.027/frame, and the positive rate of blood test in Dongzhuang Village has steadily dropped from 5.93% to 3.74%. Conclusions The infected snails missed by microscopy were detected by LAMP in Hannan District, which created conditions for the rapid emergency treatment of environment and elimination of positive snail and improved the sensitivity of the surveillance and early warning system in transmission-interrupted areas.

OBJECTIVETo investigate the mutation in mitochondrial DNA displacement-loop (mtDNA D-loop) region in oncocytoma and its relationship with tumorigenesis and tumor development.

METHODSThe mtDNA D-Loop region of 20 thyroid or renal oncocytomas and the adjacent normal tissues were amplified by PCR, and then sequenced. Five human fetal renal tissues were collected as matched controls.

RESULTSAmong the 20 oncocytomas, 21 mutations which focused on hypervariable region I (HVI) were found in 7 tumor tissues and 1 normal tissue with the mutation rates of 35% and 5%, respectively. At the same time, 191 polymorphisms were found in the 20 cases.

CONCLUSIONmtDNA D-loop region, especially HV I, is the mutational hotspot of oncocytomas, which may be closely related with mtDNA duplicating rate and the function of mitochondria.

Adenoma, Oxyphilic ; genetics ; Base Sequence ; DNA Mutational Analysis ; DNA, Mitochondrial ; genetics ; Female ; Humans ; Kidney Neoplasms ; genetics ; Male ; Middle Aged ; Mitochondria ; genetics ; Mutation ; Polymorphism, Genetic ; Thyroid Neoplasms ; genetics

OBJECTIVETo investigate whether the alterations of microtubule and microfilament expression are responsible for the neurotoxicity of carbon disulfide.

METHODSWistar rats were administered with carbon disulfide by gavage at a dosage of 300 or 500 mg/kg for continuous 12 weeks (five times per week). Spinal cords of carbon disulfide-intoxicated rats and their age-matched controls were Triton-extracted and ultracentrifuged to yield a pellet and a corresponding supernatant fraction. Then, the contents of alpha-tubulin, beta-tubulin, and beta-actin in both fractions were determined by immunoblotting. In the meantime, their mRNA levels in spinal cords were quantified using reverse transcriptase-polymerase chain reaction (RT-PCR).

RESULTSIn the supernatant fraction, the contents of beta-tubulin and beta-actin in both treated groups increased significantly (P < 0.01) the content of beta-tubulin increased by 141% and 158% respectively, and the content of beta-actin increased by 19% and 32% respectively. In the pellet fraction, the content of beta-tubulin in both groups increased by 107%(P < 0.01) and 118%(P < 0.01) respectively, and the others keep unaffected. In the meantime, the levels of of mRNA expression of beta-tubulin and beta-actin gene were elevated consistently in CS(2)-treated groups (P < 0.01) the levels of mRNA expression of beta-tubulin increased by 207% and 212% respectively, and the levels of mRNA expression of beta-actin increased by 94% and 91% respectively.

CONCLUSIONCarbon disulfide intoxication results in alternations of microtubule and microfilament expression, and the alternations might be related to its neurotoxicity.

Actins ; genetics ; metabolism ; Animals ; Carbon Disulfide ; poisoning ; Disease Models, Animal ; Male ; RNA, Messenger ; genetics ; Rats ; Rats, Wistar ; Spinal Cord ; drug effects ; metabolism ; Tubulin ; genetics ; metabolism

OBJECTIVESThe mutations of growth hormone receptor (GHR) gene results in growth hormone insensitivity (Laron syndrome) or partial growth hormone insensitivity. This study aimed to understand the relation between mutations of GHR gene and short stature with non-growth-hormone deficiency, and the clinical feature of the patients with the GHR gene mutations.

METHODS(1) Forty-seven patients with non-growth-hormone deficiency and short stature were enrolled in this study, 33 were male and 14 female. The age of the patients were at a range of 2 - 16 years. (2) The mutations of GHR gene were identified by PCR-SSCP and DNA sequencing. (3) The characteristics of the GHR mutation was assumed by screening for the same mutations in patients' family members and the control samples.

RESULTS(1) Four GHR mutations were identified in 5 patients with non-growth-hormone deficiency: H56R, G148E, IVS6-30, -31CA > TG and IVS8 + 10G > C. These mutations were located within the extracellular domain of GHR and not reported before. Five patients were the heterozygous of H56R, G148E, IVS6-30, -31CA > TG and IVS8 + 10G > C. The detection rate of mutant heterozygous individual accounted for 10.6% (5/47). The mutations were considered non-polymorphism by the GHR gene analysis in patients' family members and control samples. (2) Comparison of the amino acid sequence of different species and the position of the mutations H56R and G148E in the GHR protein structure suggested impact of the mutations on the protein function. (3) A polymorphism site was identified in exon 6 of GHR gene: G168G (GGA > GGG). The allelic frequency of G168G had no difference between the patients with non-growth-hormone deficiency and control samples but had significant difference between Chinese and Caucasian. It seems that the G168G was a polymorphism and has no relationship with the height stature. However, there was the allele diversity in different races.

CONCLUSIONThe mutations of GHR gene were detected in the patients with non-growth-hormone deficiency. Special attention should be paid clinically to its potential pathogenesis for short stature.

Adolescent ; Asian Continental Ancestry Group ; genetics ; Base Sequence ; Case-Control Studies ; Child ; Child, Preschool ; DNA Mutational Analysis ; European Continental Ancestry Group ; genetics ; Female ; Growth Disorders ; genetics ; Humans ; Male ; Molecular Sequence Data ; Mutation ; Polymorphism, Genetic ; Receptors, Somatotropin ; genetics

OBJECTIVETo investigate the effect of tanshinone IIA on HL-60 and K562 cells apoptosis, and to assay the inhibition of the telomerase activities in the leukemia cell apoptosis induced by Tanshinone.

METHODUsing the techniques of cell culture in vitro, flow cytometry and PCR-TRAP observed the telomerase activities and apoptosis of HL-60 and K562 cells which treated by Tan IIA.

RESULT0.5 microg x mL(-1) Tan IIA could obviously inhibit HL-60 and K562 cell lines growth (P < 0.05), down-regulate c-myc, bcl-2 gene and up-regulate c-fos and p53 gene expression as well as induce leukemia cell apoptosis, the apoptotic rates of HL-60 and K562 cells were 11.8% and 21.8% respectively. The telomerase activities significant decreased, the inhibiting rates in HL60 and K562 cells were 30.8% and 50.8% respectively.

CONCLUSIONTan IIA could significantly inhibit the proliferation and telomerase activities of HL-60 and K562 cells and induce the leukemia cell apoptosis.

Apoptosis ; drug effects ; Cell Proliferation ; drug effects ; Diterpenes, Abietane ; HL-60 Cells ; Humans ; K562 Cells ; Phenanthrenes ; isolation & purification ; pharmacology ; Plants, Medicinal ; chemistry ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Proto-Oncogene Proteins c-fos ; metabolism ; Proto-Oncogene Proteins c-myc ; metabolism ; Salvia miltiorrhiza ; chemistry ; Telomerase ; metabolism ; Tumor Suppressor Protein p53 ; metabolism

OBJECTIVETo observe and compare the protective effect of garlic oil against carbon tetrachloride (CCL)-induced acute liver injury.

METHODSThe experiments include 4 preventive groups and 2 therapeutic groups. In every preventive and therapeutic group, the mice were randomized into 6 groups with 15 each, including one negative control group, one solvent control group, one CCl4 model group and 3 garlic oil groups (25, 50, and 100 mg/kg body weight). Before given a single gavage of CCl4 (80 mg/kg), the mice were pretreated with garlic oil by gavage in preventive group 1 (30 days, once daily), preventive group 2 (5 days, once daily), preventive group 3 (ahead of 2 h, once), preventive group 4 (immediately, once) or the vehicle (corn oil, 10 ml/kg) in solvent control group. In therapeutic groups, the mice were gavaged garlic oil 2 h (once, in therapeutic 1) or for 5 days (once daily, in therapeutic 2) after administration CCl. After 24 h of the last administration, blood was collected and centrifuged at 2500 r/min at 4 degrees C for 10 min, and serum was removed to measure ALT and AST activities. The liver was dissected, weighed to calculate the liver coefficient (relative liver weight). At the same time, the liver samples were studied by histological examinations.

RESULTSCompared with negative group, the liver coefficient and the activities of ALT and AST in serum of model group were increased remarkably (P < 0.01). Compared with CCl model group, the liver coefficient and the activities of ALT and AST in serum were decreased significantly (P < 0.01) by garlic oil dose-dependently in each preventive group. Simultaneously, histological assessment showed that garlic oil effectively alleviated hepatocyte injuries induced by CCl4. Comparing the preventive effects of garlic oil in every group, it was better in preventive group 3 than others. However, all indexes and histological examinations in therapeutic group 1 did not show the difference with those of CCl4 model group. In therapeutic group 2, all indexes recovered after 5 d of CCl4 administration.

CONCLUSIONSGarlic oil can prevent acute liver injury induced by CCl4 and the effect is better in ahead of 2 h group than others.

Alanine Transaminase ; metabolism ; Animals ; Aspartate Aminotransferases ; metabolism ; Carbon Tetrachloride Poisoning ; drug therapy ; prevention & control ; Chemical and Drug Induced Liver Injury ; drug therapy ; prevention & control ; Garlic ; Liver ; metabolism ; Male ; Mice ; Mice, Inbred Strains ; Plant Oils ; administration & dosage ; therapeutic use

OBJECTIVE: To determine the sequence of gene for encoding beta-lactamase produced by Klebsiella pneumoniae E3 isolated from Jiaxing Area in Zhejiang Province. METHODS The Klebsiella pneumoniae strain E3 was identified as an ESBLs-producing bacterium by inhibitor-potentiated broth dilution test. The gene encoding gamma-lactamase of the strain was amplified by PCR. The purified PCR product was cloned and sequenced by Sanger's dideoxy chain termination composition method. RESULTS The Klebsiella pneumoniae strain E3 produced both TEM and SHV gamma lactamases. The SHV encoding gene had 812 nucleotide residues responsible for encoding SHV-11 gamma-lactamase and the TEM encoding gene had 973 nucleotide residues responsible for encoding TEM-1 gamma-lactamase. CONCLUSION The Klebsiella pneumoniae strain E3 isolated from a patient in Jiaxing Area in Zhejiang Province is able to produce both TEM-1 and SHV-11 gamma-lactamases.

Objective To investigate the status of Ehrlichia (E.)chaffeensis and A naplasma (A.) phagocytophilum infection among farming populations and domestic animals in the rural area of Beijing,China.Methods Blood samples from 562 farmers and 163 blood samples including 90 goats,71 ox and 2 dogs,were collected.Specificity of IgG antibodies against E.chaffeensis and A.phagocytophilum were tested by micro-indirect immunofluorescent assay (mIFA).16S rRNA genes of A.phagocytophilum were amplified from the domestic animal blood samples and products were sequenced and analyzed by nested PCR.Results The positive rates of E.chaffeensis and A.phagocytophilum antibody were 16.5％ and 14.0％ respectively for farmers.The total positive rates of A.phagocytophilum were 2.3％ and 0 for both goats and oxen respectively.No antibody was found for the 2 tested dogs.The PCR positive rates were 48.9％ and 23.9％ for goats and oxen respectively.Three dominant varieties of A.phagocytophilum were demonstrated in goats and oxen.Conclusion The prevalence rates of E.chaffeensis and A.phagocytophilum were identified in the rural areas of Beijing.

Objective To investigate the diagnostic value of Astograph methacholine provocation test in patients with chest tightness variant asthma ( CTVA)??Methods From January 2011 to February 2017,156 patients with CTVA in outpatient or inpatient department of respiratory medicine of Kailuan General Hospital affiliated to North China University of Science and Technology were selected as case group ( chest tightness variant asthma group )??The control group were 361 non?asthmatic patients including interstitial lung disease ( 23 cases), coronary disease ( 157 cases), hypertensive cardiopathy ( 22 cases), myocardiosis (16 cases),congenital heart disease ( 3 cases),rheumatic valvular heart disease (6 cases), central airway disease (3 cases),thyromegaly (10 cases),mediastinal tumor (5 cases),thoracic or spinal deformity (8 cases),phrenoparalysis (2 cases) and vegetative nerve functional disturbance (106 cases)??All participants received pulmonay ventilation test, average daily and nightly variation rate of PEF ( Peak expiratory flow) or PEF weekly variability, Astograph methacholine provocation test ( forced expirataory volume in one second≥70% expectation),and other relevant examinations??The diagnostic value of Astograph methacholine provocation test on CTVA was assessed by analyzing the sensitivity, specificity, positive predictive value,negative predictive value,and Yunden index of Astograph methacholine airway??Results Compared with the control group (( 1??18 ± 0??44)%), theforced expiratory flow from 75% of Forced vital capcacity ( FEF75 ) index of CTVA group (( 1??29 ± 0??50 )%) had significant difference (, t＝ 2??96, P＝0??006)??The sensitivity,specificity,positive predictive value,negative predictive value,Yunden index,and diagnostic accuracy of Astograph methacholine provocation test on CTVA were 0??814,0??695,0??536,0??305, 0??509 and 0??731, respectively??Conclusion The sensitivity, negative predictive value, Yunden index and diagnostic accuracy of Astograph methacholine provocation test on CTVA were higher,whereas the specificity and positive predictive value were relatively lower,suggesting that Astograph methacholine provocation test had a reliable diagnostic value on CTVA,with lower false negative and higher false positive??

OBJECTIVETo identify the activity of hammerhead ribozyme against transforming growth factor beta1 (TGFbeta1) in a cell-free system and in activated hepatic stellate cells (HSCs).

METHODSThe ribozyme against TGFb1 was designed with computer software. The transcripts of ribozyme, disabled ribozyme and target RNAs were prepared using the RiboMAX large scale RNA production system. The in vitro cleavage reactions were performed through incubation of 32P-labeled target RNAs with ribozyme or disabled ribozyme in different conditions. The eukaryotic expression vector encoding ribozyme and disabled ribozyme were constructed, and then transfected into HSC-T6 cells which exhibited characteristics of activated HSCs. The intracellular activity of the ribozyme was determined by detecting the ribozyme, disabled ribozyme and the TGFbeta1 expression.

RESULTSThe ribozyme cleaved target RNAs into anticipated products effectively. As expected, the disabled ribozyme possessed no cleavage activity in vitro. Further study demonstrated that the ribozyme expressed efficiently and inhibited TGFbeta1 expression in activated HSCs, while the disabled ribozyme displayed only a slight effect on TGFbeta1 expression.

CONCLUSIONThe ribozyme with perfect cleavage activity in the cell-free system used inhibited TGFbeta1 expression effectively in activated HSCs. This ribozyme can provide a potential therapeutic approach for liver fibrosis.

Animals ; Cell-Free System ; Cells, Cultured ; Genetic Vectors ; Hepatocytes ; cytology ; RNA ; genetics ; metabolism ; RNA, Catalytic ; RNA, Messenger ; genetics ; metabolism ; Rats ; Transcriptional Activation ; Transfection ; Transforming Growth Factor beta ; genetics ; metabolism