Salmonella enterica serovar Cerro 87, which was isolated from a commercial egg-producing farm, has a phosphorothioated DNA backbone resulting DNA degradation(Dnd) during the pulsed field gel electrophoresis (PFGE) process. In this research, a gene deletion mutant XTG103 was engineered with the entire dnd gene cluster knocked out by double crossover using vector pKOV-kan, and lost Dnd phenotype corre- spondingly. We regulated the DNA phosphorothioation by heterogenous expression of dnd gene cluster with an isopropyl ?-D-1-thiogalactopyranoside (IPTG) inducible promoter PlacZ.

Objective To evaluate the effectiveness of intratracheal injection of pulmonary surfactant (LISA) combined with continuous positive airway pressure (CPAP) on the treatment of neonatal respiratory distress syndrome (RDS). Methods The databases of PubMed, EMBASE, Cochrane, China National Knowledge Infrastructure (CNKI), Wanfang Database, Weipu Database and China Biology Medicine disc (CBM) were searched, and all of the randomized controlled trial of LISA combined with CPAP for treatment of RDS were included. RevMan 5.2 was used to perform meta-analysis. The differences of mortality, bronchopulmonary dysplasia (BPD), retinopathy of prematurity (ROP), intraventricular hemorrhage (IVH), periventricular leukomalacia (PVL), and other complications between LISA group and control group were compared in <34 weeks gestational age preterm infants with RDS. Results Five randomized controlled trials were included with 759 infants in total. Compared with control group, the rate of mechanical ventilation (RR: 0.32,95% CI: 0.13-0.82) and the incidence of BPD (RR: 0.61,95%CI: 0.42-0.88) were significantly decreased in LISA group. There were no differences in mortality and the incidences of ROP, IVH, and PVL between two groups (P>0.05). Conclusion The mechanical ventilation and the incidence of BPD is significantly reduced in the treatment of neonatal respiratory distress syndrome by LISA, but it has no effect on the incidences of other complications and mortality.

AIM:To investigate the effects of glucagon-like peptide-1 (GLP-1) receptor agonist exendin-4 on white adipose tissue (WAT) and the underlying mechanisms.METHODS:Male C57BL/6J mice (8 weeks) were chal-lenged by high-fat diet for 12 weeks, and were randomly divided into saline group and exendin-4 group.The mRNA expres-sion of sirtuin 1 (SIRT1), adipose triglyceride lipase (ATGL), TNF-αand adiponectin of WAT was detected by real-time PCR.3T3-L1 adipocytes or mouse embryonic fibroblasts cells were treated with exendin-4 for 24 h.The protein levels of SIRT1, ATGL and hormone-sensitive lipase (HSL) were determined by Western blot.RESULTS:Exendin-4 significantly decreased epididymal fat weight, fasting blood glucose and serum triglyceride levels ( P<0.05) , and reduced body weight and serum TNF-αlevel.The mRNA expression of SIRT1, ATGL and adiponectin in WAT was all significantly up-regulated by exendin-4, which were contrary to the down-regulation of TNF-αmRNA expression (P<0.05).Exendin-4 promoted the protein expression of SIRT1, ATGL, and HSL in 3T3-L1 adipocytes in a dose-dependent manner.Less lipid droplets with up-regulation of lipolytic protein expression were observed when combined with SIRT1 agonist treatment, which were suppressed by SIRT1 inhibitor.Deletion of SIRT1 led to larger adipocytes with more lipid droplets, and the effect of ex-endin-4 on the lipolysis disappeared when SIRT1 was deficient.CONCLUSION:Exendin-4 promotes lipolysis in WAT of obese mice via activation of SIRT1.

Objective TostudytheinfluenceofmeteorologicalconditionschangingontheactivityintensityofinfluenzaA (H1 N1 )and to establish the prediction model of H1 N1 positive rate associated with meteorological factors.Methods TheinfluenzaA(H1N1)datafromAprilof2009toJanuaryof2011insentinelhospitalanddailymeteorologicaldatain the same period were collected,which were analyzed by Spearman correlation analysis.The prediction model was establishedusingChi-squaredautomaticinteractiondetector(CHAID).Results Weeklyaverageatmospheric pressure(r =0.50),highest atmospheric pressure (r =0.51 ),lowest atmospheric pressure (r =0.50),average temperature (r=-0.40),highest atmospheric temperature(r=-0.41),lowest atmospheric temperature(r=-0.39), precipitation(r=-0.23 )and average wind speed (r=-0.22 )were positively correlated with the activity intensity of H1N1(all P<0.05).Factors that affected H1N1 positive rate were lowest atmospheric pressure,average wind speed and precipitation(P<0.05 ).The prediction model of H1 N1 positive rate showed that the correct rate of prediction was 66.67%.Conclusion Lowestatmosphericpressure,averagewindspeedandprecipitationarecloselyassociatedwith the activity intensity of influenza A (H1 N1 ).CHAID method can be used to predict the H1 N1 epidemics.

Objective To analyze the clinical speciality of invasive fungal infection(IFI)and provide doctors with clinical evidence for early anti-fungal therapy.Method One hundred and thirty-seven patients with 91 male and 46 female,who suffered from invasive fungal infection in ICU from January.1,2000 to June 30, 2006,were enrolled in this study.The age ranged from 17 to 82 years old.Out of 137 patients with IFI,the percentage of albicans candida,glabirate candida,tropicalis candida and parapsilosis candida were 47.4%, 26.3%,20.4% and 3.6%,reseparately.The sputum,urine,blood and other drainages were collected to perform the fungal examination after three days of admission every three days.Results Of 137 patients,42 of them were complicated with hemorrhage,53 patients with IFI developed candida anthema in the chest,abdomen and extremity.,49 patients suffering from IFI had organ dysfunction.The chest image revealed that infiltration caused by IFI especially occurred in apex of lung in some patients.The pathogen analysis displayed that albicans candidiasis easily developed candida anthema,glabirate candidiasis frequently resulted in organ dysfunction,and tropicalis candida led to hemorrhage in some organs.Conclusions The clinical specialty,of IFI caused by candida included hemorrhage,candida anthema,organ dysfunction,and infiltration in apex of lung.

OBJECTIVETo recognize the importance of analyzing the result of immunohistochemical staining correctly.

METHODReview of the three misdiagnosed cases lymphoma and exploring the causes of misdiagnosis through reviewing their clinics, histopathology and immunohistochemistry.

RESULTSCase 1 of lymphocyte rich classical Hodgkin's lymphoma (LRCHL) was misdiagnosed as follicular lymphoma (FL) initially, the RS cells were overlooked morphologically and wrongly determined BCL-2 and CD20-positive cells as tumor cells immunohistochemically; also once misdiagnosed as nodular lymphocyte predominant Hodgkin's lymphoma (NLPHL) because the CD20-negative RS misjudged cells as the positives. Case 2 of AML tumor cells expressed TdT, CD7 and CD43 unspecifically, which misdiagnosed as T-cell lymphoblastic lymphoma (T-LBL). Case 3 of type B1 thymoma was misdiagnosed as T-LBL, because CK wasn't expressed satisfactorily resulting in neglecting neoplastic epithelial cells, and lymphocytes in the background were TdT and CD99-positive.

CONCLUSIONThe diagnosis of lymphoma should be based on morphology, immunohistochemistry, clinics, and genetics. Moreover, the correct judgment of immunohistochemical staining is essential to make right diagnosis.

Adult ; Diagnostic Errors ; Female ; Humans ; Immunohistochemistry ; Lymphoma ; diagnosis ; Male ; Middle Aged

This study was purposed to investigate the regulatory effects of differentiating mesenchymal stem cells (MSC) on osteoclast formation. The MSC from mouse compact bones were cultured and induced into osteoblasts and adipocytes for one week. To test their regulatory effect on osteoclastogenesis, osteogenically differentiated and adipogenically differentiated MSC were co-cultured with CD11b(+) monocytes and osteoclasts were identified with in situ tartrate-resistant acid phosphatase (TRAP) staining. The results showed that differentiated MSC supported osteoclastogenesis but the osteoclast supporting capacity of osteogenically differentiated MSC decreased as compared with undifferentiated MSC. More interestingly, the adipogenically differentiated MSC significantly promoted osteoclasts formation when co-cultured with monocytes. It is concluded that the regulatory effect of MSC on osteoclast formation has changed while they have differentiated into different types of cells. The findings indicate that MSC may exert alternative effect on osteoclastogenesis by differentiation to descendant cells.
Adipogenesis ; Animals ; Bone Marrow Cells ; cytology ; Cell Differentiation ; Cells, Cultured ; Coculture Techniques ; Mesenchymal Stromal Cells ; cytology ; Mice ; Mice, Inbred BALB C ; Mice, Inbred C57BL ; Monocytes ; cytology ; Osteoblasts ; cytology ; Osteoclasts ; cytology

OBJECTIVETo explore the effects of ICAM-1 gene transfection on the differentiation of MSCs to adipocytes.

METHODSThe recombinant retroviral expression plasmid MIGR1-ICAM-1 containing full length of mouse ICAM-1 gene was constructed. The constructed plasmid MIGR1-ICAM-1, empty plasmid MIGR1 and packaging plasmid ECOS were transfected into T293 cell lines and then the supernatant generated from T293 cells were used to infect mouse MSCs cell line C3H10T 1/2. The transfective efficiency was determined by inverted fluorescence microscope, real-time PCR and flow cytometry. Furthermore, ICAM-1 overexpressing MSCs (C3H10T 1/2-ICAM-1) and empty vector transfection MSCs (C3H10T 1/2-MIGR1) were cultured in medium with or without induction reagents, Oil-red-O staining was used to detect the lipid accumulation, and the expression of transcriptional factors C/EBPα and PPARγ, which were key factors in the differentiation of MSCs to adipocytes, were tested by real-time-PCR.

RESULTSThe recombinant retrovirus vector containing mouse ICAM-1 gene was successful constructed. After transfection into MSCs cell line C3H10T 1/2, the overexpression ICAM-1 MSCs cell line (C3H10T 1/2-ICAM-1) and control cell line (C3H10T 1/2-MIGR1) were obtained. Furthermore, these two cell lines were treated without or with adipocytic induction reagents, C3H10T 1/2-ICAM-1 showed significantly lower mRNA expression level for C/EBPα [(1.2 ± 0.7), (2.9 ± 0.9)] and PPARγ [(1557.6 ± 70.2), (7547.0 ± 442.2)] when compared with C3H10T 1/2-MIGR1 [(5.8 ± 0.5), (23.0 ± 2.3) and (2453.0 ± 215.6), (9856.3 ± 542.2)](P < 0.05). Moreover, little lipid droplet and decreased quantity of adipocytes were detected in C3H10T 1/2-ICAM-1 [(3.2 ± 0.5)/well, (12.2 ± 3.8)/well] than that in C3H10T 1/2-MIGR1 [(11.2 ± 0.4)/well, (51.3 ± 2.8)/well] (P < 0.05).

CONCLUSIONOverexpression of ICAM-1 in MSCs can inhibit its adipocytic differentiation.

Adipocytes ; cytology ; Animals ; Cell Differentiation ; Cell Line ; Intercellular Adhesion Molecule-1 ; genetics ; Mesenchymal Stromal Cells ; cytology ; Mice ; Transfection

OBJECTIVETo investigate HBV mutations in reverse transcriptase (RT) gene and precore/basal core promoter (PC/BCP) regions in a chronic hepatitis B patient and to analyze the link between the mutations and drug resistance or HBeAg sero-conversion.

METHODSEighteen serum samples were collected from a chronic hepatitis B patient during his 14 hospitalizations from June 2002 to September 2007. HBV DNA was extracted and nested PCR was employed for amplification of target gene fragments. Direct sequencing of PCR products was performed followed by analysis with NTI software. The significance of the results was analyzed in combination with the clinical data of the patient.

RESULTSSeveral mutations were identified in succession, including LAM-resistant mutations M204I/V and L180M+M204V, ETV-resistant mutation S202G, and HBeAg nonsense mutation G1896A. The results were in accordance with the disease progression of the patient.

CONCLUSIONSequencing of HBV RT and PC/BCP regions is valuable for comprehensively checking the viral mutations and thus it is helpful in the surveillance of patients in clinics as a way for adopting reasonable antiviral therapy.

Adult ; Antiviral Agents ; pharmacology ; DNA, Viral ; genetics ; Drug Resistance, Multiple, Viral ; genetics ; Genotype ; Hepatitis B virus ; genetics ; Hepatitis B, Chronic ; virology ; Humans ; Male ; Mutation

Adolescent ; Adult ; Agranulocytosis ; microbiology ; Anti-Bacterial Agents ; adverse effects ; therapeutic use ; Daptomycin ; adverse effects ; therapeutic use ; Female ; Gram-Positive Bacterial Infections ; drug therapy ; Humans ; Male ; Middle Aged ; Young Adult