A pathogenic bacterial strain X1 was isolated from Siberian sturgeon (Acipenser baerii) suffering with bacterial septicemia. The 50% lethal concentration (LC50) of strain X1 was 5.62?105 CFU/mL, which showed that strain X1 was rather strong virulent to Acipenser baerii. Strain X1 was gram negative and 1.0 ?m～1.2 ?m ? 2.1 ?m～2.4 ?m in size with peritrichous flagella, and had ?-hemolytic activity on rabbit blood agar. By means of ATB expression identification and 16S rDNA sequence analysis, strain X1 was identified as Aeromonas hydrophila (locus number: EU669667), which was the closest relative to Aeromo-nas hydrophila strain ATCC35654 (locus number: X74676.1) with 99% homology. In addition, strain X1 was highly sensitive to cefoperazone and cravit, and intermediately sensitive to ten kinds of antibiotics in-cluding tobramycin, norfloxacin, sulperazone, kanamycin, gentamycin, fortum, vancomycin, neomycina, polymyxin B and lomefloxacin.

Objective Respiratory syncytial virus (RSV) is the most common cause of lower respiratory infection in infants.It is very important to quantitative assay of RSV titer in the study on RSV pathogenesis,candidate vaccine and antiviral treatment.Therefore,we develop Real-time Quantitative PCR (Q-PCR) assay and enzyme immunospots (EIS) for titrating RSV and compare them with traditional 50% tissue culture infectious doses (TCID_(50)).Methods Q-PCR,based upon the RSV-L genes,and EIS were utilized to titrate samples from RSV culture supernatants and RSV infected mouse lungs.Then,the results were compared with TCID_(50).Results For the samples from RSV culture supernatants,the ratio of Q-PCR and EIS (plaque forming unit,pfu) was 10:1 and the ratio of EIS and TCID_(50) was 10:1 when TCID_(50) was converted as pfu.For the samples from RSV infected mouse lungs,the ratio of Q-PCR and EIS was 1000:1 and the ratio of EIS and TCID_(50) was 5:1.Conclusion We have successfully established Q-PCR and EIS to titrate RSV and compared them with TCID_(50).We concluded EIS is a cost-effective method to titrate RSV.

Objective To explore the risk factors of hyperbilirubinemia in late preterm infants. Methods Clinical data of 211 cases of late preterm infants with hyperbilirubinemia and 246 cases of late preterm infants without hyperbilirubinemia were retro-spectively analyzed between 2011 and 2012. The risk factors of hyperbilirubinemia were filtered. Results Twenty-seven cases of late premature infants with hyperbilirubinemia were severe. Hospital stay less than 3 days, birth asphyxia history, small for gestatio-nal age, head hematoma, delivery injury, hypoalbuminemia, polycythemia, infection, hemolytic disease, feeding intolerance, and fe-tal excretion delay were associated with hyperbilirubinemia (P<0.05). Rural origin, pregnancy-induced hypertension syndrome and premature rupture of membrane were also associated with hyperbilirubinemia (P<0.05). Multivariate logistic regression analysis showed the history of birth asphyxia , fetal excretion delay, hypoalbuminemia, pregnancy-induced hypertension syndrome were risk factors of hyperbilirubinemia in late preterm infants (OR=2.35-4.05). Pregnancy-induced hypertension syndrome and hemolytic dis-ease were risk factors of severe hyperbilirubinemia in late preterm infants (OR=5.74, 73.64). Conclusions Neonatal asphyxia, fetal excretion delay, hypoalbuminemia and pregnancy-induced hypertension syndrome are risk factors of hyperbilirubinemia in late pre-term infants. Strengthening the management of pregnancy-induced hypertension syndrome and the treatment of newborn hemolytic disease can reduce the occurrence of severe hyperbilirubinemia in late preterm infants.

Objective Through clinical observation and statistics, to get the best curative effect of surgical operation in treating pigment nevus method and provide clinical guidance. Methods We reviewed of face and neck patients (1100 patients) with pigmented nevus in the department in the department of the dermatology,plastic surgery from January 2013 to October 2015, two different methods was designed on each parts and effect of the treatment, especially satisfaction degree was analyzed by statistical methods.Results In 100 cases of patients, only 20 patients had mild scar hyperplasia at the neck incision and the rest of the patients were satisfactory. For special parts such as mouth,nose and eye around,along thedirection of the muscle, arc and along the direction of dermatoglyph incision was designed respectively, patients obtained with higher postoperative satisfaction (P<0.05) . Conclusion In pigmented nevus of face and neck surgery treatment, surgical incision design requires dynamic and static combining method,incision design is important for the postoperative effect and patients' satisfaction.

Objective To detect the expression and content of decorin in fibroblasts of keloid to deeply reveal the mechenism and the role of decorin plays in scar formation.Methods Fibroblasts of keloid,normal scar and normal skin were cultured in vitro,and the morphology,activity,apoptosis of fibroblast were observed under light microscope and electron microscope; the mRNAs of decorin and TGF-β1 were detected and analyzed with real-time fluorescent quantitative-PCR (FQ-PCR).Results Fibroblasts of keloid showed irregular morphology,larger size and disorder arrangement.There were a large number of mitochondria,swelling rough endoplasmic reticulum,and euchromatin-rich in nucleus of fibroblasts,suggesting the protein synthesis of keloid fibroblast was very active.Compared with normal skin,the expression of decorin was significantly lower in keloid fibroblast; On the contrary,the expression of TGF-β1 was significantly higher in keloid fibroblast than in normal scar and normal skin.Conclusions Compared with normal skin,the expression of decorin in keloid fibroblast is significantly lower.Lower content of decorin in early stage of wound healing may induce weakly suppression of proliferation and synthesis of fibroblast,and up-regulate the activity of TGF-β1,which promotes the proliferation,migration and excessive collagen synthesis of the fibroblast of keloid.Thus,decorinis an suppressor factor of keloid formation.

BACKGROUND: The gut is capable of inducing multiple organ dysfunction syndrome (MODS). In the diagnosis and treatment of critical ill patients, doctors should pay particular attention to the protection or recovery of intestinal barrier function. However, no reliable diagnostic criteria are available clinically. This study aimed to assess the changes of intestinal mucosal barrier function in surgically critical ill patients as well as their significance. METHODS: Thirty-eight surgically critical ill patients were enrolled as a study group (APACHE II>8 scores), and 15 non-critical ill patients without intestinal dysfunction were selected as a control group (APACHE II<6). General information, symptoms, physical signs, and APACHE II scores of the patients were recorded. The patients in the study group were subdivided into an intestinal dysfunction group (n=26) and a non-intestinal dysfunction group (n=12). Three ml venous blood was collected from the control group on admission and the same volume of plasma was collected from the study group both on admission and in the period of recovery. The plasma concentrations of endotoxin, diamine oxidase (DAO), D-lactate, and intestinal fatty-acid binding protein (iFABP) were detected respectively. The data collected were analyzed by the SPSS 17.0 software for Windows. RESULTS: The levels of variables were significantly higher in the study group than in the control group (P<0.01). They were higher in the intestinal dysfunction group than in the non-intestinal dysfunction group (DAO P<0.05, endotoxin, D-lactate, iFABP P<0.01). In the non-intestinal dysfunction group compared with the control group, the level of endotoxin was not significant (P>0.05), but the levels of DAO, D-lactate and iFABP were statistically significant (P<0.05). The levels of variables in acute stage were higher than those in recovery stage (P<0.01).The death group showed higher levels of variables than the survival group (endotoxin and D-lactate P<0.01, DAO and iFABP P<0.05). CONCLUSION: The plasma concentrations of endotoxin, DAO, D-lactate, and intestinal fatty-acid binding protein (iFABP) could reflect a better function of the intestinal mucosa barrier in surgically critical ill patients.

Background and Objective: Monitoring the therapeutic effects of radiotherapy for nasopharyngeal carcinoma(NPC)is critical to providing individualized treatment.This in-vivo study was initially designed to evaluate the therapeutic effect of radiotherapy using 18F-fluorodeoxyglucose positron emission tomography with computed tomography(18F-FDG PET-CT)imaging.Methods: 18F-FDG PET-CT imaging was performed on all of the 10nude mice bearing NPC xenografts before radiotherapy,and early-phase and delayed-phase PET-CT images were performed on 7 of the 10 mice.All mice were randomly divided into either a control group or a radiotherapy group.The5 mice in the control group were immediately killed after the imaging and pathology were performed.After receiving radiotherapy of 12 Gy,5 animals in the radiotherapy group were given 18F-FDG PET-CT imaging on days 2,4,and6,and then were killed for pathologic evaluation.Regions of interest(ROI)technology was used to measure the tumor target/non-target(T/NT)ratio and the volume of the tumors.Results: The average T/NT ratios of early-and delayed-phase imaging were 1.806±0.532 and 1.777±0.597,respectively,with no significance(P>0.05).For the radiotherapy group,the average T/NT ratios for 18F-FDG PET-CT before radiotherapy,and on days 2,4,and 6after radiotherapy,were 1.735±0.466,1.818±0.396,1.096±0.101,and 0.604±0.108,respectively,and the tumor volumes were(1.48±0.27)cm3,(1.57±0.31)cm3,(1.59±0.31)cm3,and(1.60±0.29)cm3,respectively.The average T/NT ratios of day 6 after radiotherapy and the other time points were significantly different(P<0.05).The average death ratio of the tumor cells was(93.00±7.42)% after 6 days of post-radiotherapy.Conclusions: 18F-FDG PET-CT imaging can be used for the early assessment of radiotherapeutic effect of NPC in vivo.Day 6 after radiotherapy may be an appropriate time point for the imaging.However,the T/NT ratio measurement of delayed-phase imaging might make no sense for the diagnosis of NPC.

OBJECTIVETo construct a lentiviral expression vector for short hairpin RNA (shRNA) of human survivin gene, and assess its gene silencing effect in human ectopic endometrial cells.

METHODSHuman survivin gene shRNA sequence was designed using a software available on-line. The synthesized shRNA sequence was cloned into the pGCL-GFP vector to construct LV-survivin shRNA, which was confirmed by PCR and DNA sequence analysis. The packaging 293T cells were cotransfected with LV-survivin shRNA, pHelper 1.0 and pHelper 2.0, and the titer of the lentivirus was determined. The recombinant lentivirus was injected into human ectopic endometrial cells and the survivin mRNA expression was examined by semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR) in comparison with that in the non-transfected and blank vector-transfected human ectopic endometrial cells.

RESULTSPCR analysis and DNA sequencing confirmed correct insertion of the shRNA sequence into the lentiviral vector. The titer of virus after packaging was 8x10(8) U/ml. Survivin mRNA expression in human ectopic endometrial cells transfected by LV-survivin shRNA was significantly inhibited compared with those in the non-transfected and empty vector transfected human ectopic endometrial cells (P<0.01), and no significant difference was found between the latter two groups.

CONCLUSIONThe lentiviral shRNA vector of survivin gene constructed can effectively inhibit the expression of survivin gene in human ectopic endometrial cells in vitro. This vector provides a tool for investigating the role of survivin gene in the occurrence and progression of endometriosis and for searching new therapeutic targets.

Cells, Cultured ; Endometriosis ; genetics ; pathology ; Female ; Gene Targeting ; Genetic Vectors ; genetics ; Humans ; Inhibitor of Apoptosis Proteins ; biosynthesis ; genetics ; Lentivirus ; genetics ; metabolism ; RNA Interference ; RNA, Messenger ; biosynthesis ; genetics ; RNA, Small Interfering ; genetics ; Recombinant Proteins ; biosynthesis ; genetics ; Transfection

Animals ; Disease Models, Animal ; Liver ; parasitology ; Mice ; Mice, Inbred BALB C ; Schistosomiasis japonica ; immunology ; Thiourea ; analogs & derivatives ; pharmacology

OBJECTIVEThis study investigated the role and mechanism of calcineurin (CaN)-nuclear factor of activated T cells (NFAT) pathway in the myoblast apoptosis induced by cyclic tensile strain.

METHODSMyoblasts were cultured using an in vitro-mechanical stimulation model and imposed with tension for different hours with a multi-channel cell stress loading system. Cyclosporine (CsA) was used as CaN inhibitor to clarify the role of CaN in the apoptosis induced by cyclic stress. Hochest 33258 staining and flow cytometry detection were performed to detect the apoptotic cells. Real-time polymerase chain reaction was conducted to detect the mRNA expression of CaN and NFAT. Protein levels of NFAT3 were evaluated by Western blot.

RESULTSThe apoptosis rate increased with the extension of loading time. The mRNA expression of the CaN subunits, CnA and CnB, and the protein levels of NFAT3 also increased. When the myoblasts were incubated with CsA, the apoptosis rate decreased, the mRNA expression of CnA and NFAT3 significantly decreased, and the NFAT3 protein expression levels became significantly lower than those of the groups without CsA.

CONCLUSIONContinuous cyclic tensile stress can induce myoblast apoptosis. The CaN-NFAT signaling pathway may be involved in the cyclic stretch-induced apoptosis of myoblasts.

Apoptosis ; Calcineurin ; genetics ; Cyclosporine ; Flow Cytometry ; Myoblasts ; physiology ; NFATC Transcription Factors ; metabolism ; Real-Time Polymerase Chain Reaction ; Signal Transduction ; T-Lymphocytes