Objective To develop a new method for rapid detection of E.Coli.with a new substrate Na p nitrophenyl ? D glucuronate (PNP Glucu Na). Method The test paper was immersed with of Na p nitrophenyl ? D glucuronate which was synthesized by us and another paper with culture medium and phorphate buffer.Colour change was observed to detect the E.coli. Results An E.coli concerntration of 4?10 7 took 0.5 hour, while a concentration of 4?10 0 took 12 hours. 138 strains of E.coli and 190 strains of other bacteria were examined . The sensitivity was 94.2% and the specificity was 92.6%. Conclusion This method is rapid, accurate, simple, and economical for detecting E.coli.

Objective To investigate the method of establishing lymph node metastasis model for gastric cancer in nude mice.Methods Human poorly differentiated gastric cancer SGC-7901 ceils were cul- tured,inherited,amplified and collected in vitro,and were inoculated in nude mice subcu.taneously.After pas- saging six generations,the subcutaneous tumor was orthotopically implanted in nude mice.The mice were then sacrificed on the 9th week after implantation.Serum CEA density was detected using radio-immunity method. Orthotopically implanted tumors,remote organ metastases as well as lymph node metastasis were observed macrescopically and histologically.Results The successful rate of orthotopically transplantion of gastric can- cer was 100 %.The rate of perigastric lymph node metastases was 93.3 %.Local infiltration and remote metastasis were observed.The level of serum CEA in nude mice bearing tumor was significantly higher than that of normal nude mice(P

Three myelin proteins, Nogo-A, MAG and OMgp, transduce their neurite-outgrowth inhibitory signal through a common receptor complex: NgR/ p75NTR (or TROY). Recently, LINGO-1 is identified as another essential component and regulator for the Nogo-66 receptor/p75 signaling complex. LINGO-1 is restricted to express in CNS, neuronal LINGO-1 is shown to be involved in the signal transduction from three myelin proteins, and Lingo-1 in oligodendrocyte negatively regulates the differentiation and myelination of oligodendrocyte. To investigate the potential activity of LINGO-1 in neuronal apoptosis, LINGO-1-Fc fusion protein including the extracellular LRR and IgC2 domain, was used as functional antagonist to study its protective effect on low-potassium induced apoptosis of cerebellar granule neurons (CGNs). In judgement of the apoptotic nuclei stained by Hoechst, LINGO-1-Fc pretreatment for 2 h significantly prevents apoptosis of CGNs. Although GST-LINGO-1 protein, including the LRR domain, binds to the CGN cultures in the same way with LINGO-1-Fc, it doesn't prevent the apoptosis of CGNs. These results indicate that LINGO-1-Fc fusion protein prevents low-potassium induced apoptosis of cerebellar granule neurons in certain conditions and this activity is probably IgC2 domain dependent.

Objective To investigate the effect of alpha 1-antitrypsin (A1AT) concerning in reducing the injury of transplanted islets by pancreas exocrine cells and promoting proliferation of the pancreas B cells.Method The pancreases of mice were digested with collagenase,islets were isolated artificially,and pancreatic exocrine cells were collected.In purified islet group (n =6),100 islets were seeded into a 6 well culture plate.In experimental group(n =6),100 islets were co-cultured with equal volume of pancreas exocrine cells,and 0.5 mg/mL A1AT was added into a 6-well culture plate.In control group(n =6),100 islets were co-cultured with equal volume of pancreas exocrine cells.After 48 h,insulin content of islets in each well and trypsin concentration in the supernatant of each well were measured.The islets were cultured in low sugar and high sugar 1640 medium,then glucose stimulated insulin secretion (GSIS) test was carried out.In vivo,8-9-week old male BALB/C mice were induced with STZ (190 mg/kg body weight,i.p) to establish the diabetic model and randomly divided into two groups.In experimental(n =10) and control(n =10) groups,250 islets and the equal volume of pancreatic exocrine cells were transplanted into different regions of left kidney subcapsule,resepctively.The experimental group was injected with A1AT (83 mg/kg,qd,i.p) for 28 days after operation,and the control group was injected with the same amount of normal saline (qd,i.p) for 28 days.Both two groups were given EDU (5 μg/g,qd,i.p) for 28 days.The blood glucose level was monitored continually.Nephrectomies were performed after 28 days.The expression of anti-amylase antibodies in the renal subcapsule was detected by immunohistochemical staining,and the proliferation of islet beta cells was examined using immunofluorescence staining.Result Insulin levels and insulin stimulation index in the control group were decreased as compared with those in the purified islet group; those in the experimental group were higher than in the control group,but lower than in the purified islet group.Trypsin concentration in the control group was increased as compared with the purified islet group,that in the experimental group was lower than the control group,but higher than in the purified islet group (all P＜0.01).After islets transplantation,the blood glucose levels in control and experimental groups were normal,but those in the control group recovered later than in the experimental group (P＜0.01).At 3rd day after nephrectomy,the blood glucose levels were ＞21 mmol/L in both two groups.A large number of anti-amylase antibody-positive cells were found in the renal subcapsule in the control group while little seen in the experimental group after 28 days.The immunofluorescence showed that the insulin +/EDU + B cells in the experimental group were more than those in the control group.Conclusion Conclusion Co-culture of islets and pancreatic exocrine cells with A1AT can prevent islet cells from damage caused by trypsin.A1AT could inhibit the secretion of pancreatic amylase from pancreatic acinar cells and promote proliferation of islet beta cells.

Objective To observe the molecular weight distribution of soybean peptides prepared by complex enzyme hydrolysation and tumor inhibitory effect in vivo or in vitro. Method The soybean protein was enzymolyzed to soybean peptides by Flavourzyme, Alealase and neutral protease, at(50-55)℃, followed by separation, absorption,affination, ultra centrifuge and spray drying. The soybean peptides were passed Sephadex G25 column at 0.5ml/min with 0.1mol/L Tris-HCl buffer. The fragment of fluid collected was determined by UV at 280nm and HPLC. Then the molecular weight distribution was calculated. The inhibitory effect on ansplant glioma (G422) at dose 1.25, 2.5 and 5g/kg bw (ig daily for 11 d) was tested in mice and in Hep-2 in vitro. Results The peptides contents in three batches were 54.6%, 51.9%, 51.8% respeclively. The molecular weight

Exosomes are 40–100 nm nano-sized vesicles that are released from many cell types into the extracellular space. Such vesicles are widely distributed in various body fluids. Recently, mRNAs and microRNAs (miRNAs) have been identified in exosomes, which can be taken up by neighboring or distant cells and subsequently modulate recipient cells. This suggests an active sort-ing mechanism of exosomal miRNAs, since the miRNA profiles of exosomes may differ from those of the parent cells. Exosomal miRNAs play an important role in disease progression, and can stimu-late angiogenesis and facilitate metastasis in cancers. In this review, we will introduce the origin and the trafficking of exosomes between cells, display current research on the sorting mechanism of exo-somal miRNAs, and briefly describe how exosomes and their miRNAs function in recipient cells. Finally, we will discuss the potential applications of these miRNA-containing vesicles in clinical settings.

To confirm the hypothesis that the high frequency sequences of high throughput sequencing are the terminal sequences of the bacteriophage genome. An adaptor of specific sequence was linked to the end of the bacteriophage T3 genomic DNA, which was then subject to high throughput sequencing; as a control, the same T3 genomic DNA without adaptor was also analyzed by high throughput sequencing. The sequencing results were examined with bioinformatics software. Similar high throughput sequencing technique was applied to analyze the genomic sequence of N4-like bacteriophage IME11. Bioinformatics study showed that the sequences tagged with adaptors were consistent with the high frequency sequences without adaptor labeling. Our analysis also indicated that the end of the T4-like phage genome had specific sequences instead of random sequences, disagreeing with the previous assertion. Evidences were provided that N4-like bacteriophage had a particular terminal sequence: the left end of the genome was unique while the right end was permuted. The high throughput sequencing technique was convenient and practical to be used to simultaneously detect the terminal sequence and the complete sequence of bacteriophage genome.
Caudovirales ; genetics ; Computational Biology ; Genome, Viral ; High-Throughput Nucleotide Sequencing ; methods

OBJECTIVETo investigate the association between long non-coding RNAs (lncRNAs) and brain injury in inflammation-induced preterm mice, and to provide a reference for the prevention and treatment of brain injury.

METHODSAn intraperitoneal injection of lipopolysaccharide in pregnant mice was performed to establish a model of inflammation-induced preterm mice with brain injury (preterm group). The full-term mice delivered by normal pregnant mice were used as controls (full-term group). The lncRNA chip assay was used to screen out the lncRNAs associated with brain injury in preterm mice. Quantitative real-time PCR was used to validate the lncRNAs identified by the above method.

RESULTSThe preterm and full-term groups showed significant differences in the expression of 1 978 lncRNAs (P<0.05), consisting of 786 up-regulated lncRNAs and 1 192 down-regulated lncRNAs, and 29 lncRNAs were 1.5 or more times differentially expressed between the two groups. A further analysis was performed for the 10 most differentially expressed lncRNAs, and the results showed that these lncRNAs were involved in the biological processes including transcription, signal transduction, apoptosis, cell cycle, and inflammatory response, as well as G protein-coupled receptor signaling pathway and neuropeptide signaling pathway. Real-time PCR was performed to validate the expression of two lncRNAs in brain tissue in the preterm and full-term groups, and the results were consistent with those of the chip assay.

CONCLUSIONSThe expression profiles of lncRNAs in brain tissue change significantly in inflammation-induced preterm mice, and the G protein-coupled receptor signaling pathway may be involved in the pathogenesis of preterm brain injury.

Animals ; Brain ; metabolism ; Female ; Inflammation ; complications ; metabolism ; Mice ; Mice, Inbred BALB C ; RNA, Long Noncoding ; analysis ; Receptors, G-Protein-Coupled ; physiology ; Signal Transduction ; physiology

Many studies indicated that meteorological factors are associated with stroke risk,especially cold temperature.This article reviews the effect of cold temperature on the occurrence of stroke.

Objective To identify the epidemic characteristics and risk factors of an emerging infectious disease-severe fever with thrombocytopenia syndrome (SFTS) in Hubei province.Methods Active surveillance program on SFTS was set up in monitoring sites-hospitals,at the township level or above,in Suizhou,Huanggang and Wuhan from January to December,2010.Specific surveillance program on SFTS was launched across the province in hospitals above the county level.Cases that matched the definition of surveillance case were identified and reported to Centers for Disease Control and Prevention (CDCs).Cases were interviewed and their blood samples collected and detected using PCR and virus isolation.We also conducted serum antibody surveys among healthy population and livestock and surveillance on vector ticks in those high-epidemic areas.Results 188 cases that matched the definition of surveillance case and 21 deaths were reported in 11 cities,32 countries and 100 towns in 2010,with an incidence rate of 0.33/106.The fatality rate was 11.2％.Data showed that the patients were from hilly areas at the altitude elevated between 28-940 meters.The epidemic period was between April and December with the peak from May to September.The youngest case was an 11-year old,while the eldest was 81 with median age as 56-year old.95.3 ％ of the patients were farmers.All Patients did not have the history of traveling,two weeks before the onset of SFTS.93.6％ of the patients engaged in different kind of work which was associated with agriculture.52.8％ of the patients had been exposed to ticks.22.0％ of the patients had been bitten by ticks.Skin injury was found in 64.2％ of the patients.Samples from 129 cases (68.6％) were collected and detected,with 67.4％ of them (87 cases) showed positive by Real time-PCR for SFTS virus.An elevation in antibody titer by a factor of four or evidence of sero-conversion was observed in 11 patients; SFTS virus was isolated from 2 patients.The total antibody positive rates were 3.8％,55.0％ (6/11 ),36.7％ (2/3) and 80.0％ (4/5) respectively in healthy population,dogs,sheep and cows.Ticks from grass,cattle and sheep were detected positive by Real time-PCR.Conclusion Most cases of SFTS in Hubei were infected by SFTS virus,and cases of livestock were infected by SFTS virus.Ticks might serve as an important vector.Skin injury,exposure to tick bites seemed to be the risk factors.