Effect of Annexin A1 on the Cardiac Function and Inflammatory Response of Diabetic Rats / 中国医科大学学报

Objective To study the effect of annexin A1 on cardiac function,tumor necrosis factorα(TNF?α),and interleukin 1β(IL?1β)in diabetic rats. Methods Twenty?four SD rats were randomly divided into normal control and diabetic groups. The type 2 diabetes model was in?duced with a high?glucose and high?fat diet and administration of low?dose streptozotocin.Left ventricular end?diastolic volume(LVEDV),left ven?tricular end?systolic volume(LVESV),peak velocity of early diastolic mitral?to?late diastolic peak velocity(e/a)ratio,left ventricular ejection frac?tion(LVEF),and stroke volume(SV)were measured by using color Doppler ultrasonography at the end of week 8. The expression levels of TNF?αand IL?1βin blood were measured by using enzyme?linked immunosorbent assay,and the expression level of annexin A1 in blood was measured at weeks 0,4,and 8 by using real?time polymerase chain reaction. Results Compared with the normal control group,the diabetic group had de?creased LVEDV,e/a,and SV(P<0.05).The annexin A1 expression level in the diabetic group decreased significantly after 8 weeks(P<0.01). The TNF?αand IL?1βlevels in the diabetic group were significantly higher than those in the normal control group(P<0.05)and increased signifi?cantly after 8 weeks(P<0.01). Annexin A1 level correlated with the TNF?αand IL?1βlevels in the diabetic group(P<0.01). Conclusion Annexin A1 expression shows an anti?inflammatory effect that improved the cardiac function of diabetic rats.

Depletion of GP73 inhibits invasion and metastasis of hepatocellular carcinoma cells / 中华肿瘤杂志

Objective@#To explore the in vitro and in vivo effect of GP73 on the proliferation, invasion and metastasis in hepatocellular carcinoma.@*Methods@#GP73 gene was knocked out using CRISPR/Cas9 gene editing system in H22 and HepG2 cells, and stable knock out strains were constructed. The knockout efficiency was measured by western blot. Colony formation assay was used to detect the effect of GP73 on long-term survival ability. Cells were then highly synchronized in G1 phase upon treatment with cell synchronization reagents (mimosine), and the percentage of cells in G2/M phase at different time points was detected by flow cytometry. The invasive and metastasis abilities of hepatocellular carcinoma cells were detected by Transwell™ assay. Furthermore, the tumor formation abilities in vivo were examined using subcutaneous xenograft models.@*Results@#The stable knock out strains of GP73 in H22 and HepG2 cells were successfully established via puromycin selection. The number of colonies of GP73 knock out groups in HepG2 and H22 cells 10 days after transfection were 400±70 and 248±60, respectively. They were significantly lower than those in the control groups (980±40 and 1 100±50, respectively; P<0.01). In addition, GP73 knockout slowed down the cell cycle progression. Moreover, the cell numbers that had migrated to the underside of the filters were 312±50 and 305±49 in the GP73 knockout groups of HepG2 and H22 cells, respectively, significantly lower than 1 540±87 and 1 270±86 in the controls (P<0.01). For transwell invasion assay, the cell number that had invaded into the underside of the filters were 230±47 and 238±54 in the GP73 knockout groups of HepG2 and H22 cells, respectively, significantly lower than 648±74 and 596±63 in the controls(P<0.01). Furthermore, the tumor volume of GP73 knockout group was (70±170) mm3, significantly smaller than (1 200±110)mm3 of the control guoup (P<0.01).@*Conclusions@#GP73 knockout decreases the proliferation, invasive and migratory abilities of HepG2 and H22 cells in vitro and in vivo. GP73 may contribute to tumorigenesis of hepatocellular carcinoma.

Mechanism of RING finger protein 34 involved in innate immunity:a preliminary study / 军事医学

Objective To study the mechanism of RING finger protein 34 ( RNF34 ) involved in innate immunity . Methods Recombinant PCR was used and transient expression of the plasmid was achieved in HEK 293T cells.The cells were stimulated with Sendai virus ( SeV) or N-RIG-Ⅰfor the indicated time while luciferase activity was observed using the dual-luciferase reporter assay kit .Results We constructed the plasmid pcDNA 3-Flag-RNF34 and its three mutations .The study found that when stimulated by SeV , RNF34 could inhibit the activity of NF-κB and IFN-βmore significantly than RNF34-ΔFYVE, RNF34-ΔCID and RNF34-ΔRING.We also found that RNF 34 and its three mutants had similar inhibitory effect when the activation of NF-κB and IFN-βwas stimulated by the N-RIG-Ⅰ.Conclusion RNF34 negatively regulates innate immunity by acting on the RIG-Ⅰ-MAVS signaling pathway .

Construction of H22 GP73 knockout gene stable strain using CRISPR/Cas9 gene editing system and identification of functions / 军事医学

Objective To knock out the GP73 gene in H22 cells originating in mice using CRISPR/Cas9 gene editing system and construct H22 GP73 gene knockout stable strain for identification of its functions .Methods Two pairs of sgRNAs that could specifically identify the upstream and downstream of GP 73 gene first promoter were designed before a recombinant eukaryotic expressional plasmid was constructed using pX 459 .After enzyme digestion and sequencing , two pairs of recombinant plasmids were co-transfected into H22 cells before puromycin was used to screen positive cells and monoclonal cells which stably knocked out GP 73 gene were developed .The knockout effect was measured by Western blotting.Cell Titer 96? AQueous One Solution Assay was used to detect the effect on cell reproductive capacity when the GP73 was knocked out .The transferability was detected through wound healing test .Results The result of Western blotting suggested that GP73 protein was undetected in the construction of H22 GP73 knockout gene stable strain after transfection.The transfer and reproduction slowed down .Conclusion H22 GP73 gene knockout stable strain can be successfully built using CRISPR/Cas9 gene editing system ,thus facilitating studies on the function of GP 73 in hepatocarcinogenesis .

Correlational Analysis of Diastolic Heart Failure and Bone Mineral Density in the Elderly / 中国医科大学学报

Objective To study the relationship between the severity of diastolic heart failure(DHF)and bone mineral density in the elderly. Methods Totally 80 elderly patients aged over 80 years who were tested as normal for cardiac diastolic function by Doppler tissue imaging(DTI) were selected and divided into four groups by the e/a ratio,i.e.,the normal group(n=18),the DHF 1 group(0.8≤e/a<1,n=25),the DHF 2 group (0.6≤e/a<0.8,n=22),and the DHF 3 group(e/a<0.6,n=15). And the other 20 healthy people by physical examination were set as the normal control group.All subjects underwent bone mineral density(BMD)measurement(including femoral neck,total femoral hip and lumbar vertebra 1?4) by dual energy X?ray absorptiometry. Results Bone mineral density(BMD)was significantly decreased(P<0.05)in DHF groups(DHF 1,DHF 2,and DHF 3). Bone mineral density significantly decreased along with the severity of DHF. Bone mineral density was positively correlated with the e/a ratio in the elderly with DHF(r=0.75,P<0.01). Conclusion The severity of diastolic heart failure is closely related to bone mineral density in the elderly. The severity of diastolic heart failure could predict osteoporosis.

Drug Resistance and Distribution of Common Nosocomial Pathogens / 中华医院感染学杂志

OBJECTIVE To investigate the distribution of commonly encountered pathogenic microbes during the last five years.METHODS A total of 9318 strains of NI pathogens during from Jan 2004 to Dec 2008 were surveyed and analyzed.RESULTS From them the percentage of Gram-positive cocci was 13.6%,the main pathogen was coagulase negative Staphylococcus,Gram-negative bacilli(28.8%).The predominant pathogens were Escherichia coli,Klebsiella pneumoniae and Pseudomonas aeruginosa.The drug resistance of NI pathogens was markedly increased.Especially,the rate of drug resistance of P.aeruginosa to imipenem was from 0 to 31.8% in 2005,and that of Enterococcus to vancomycin was 4.0% in 2007.The percentage of fungi was 26.4% and increased sharply year by year.CONCLUSIONS The Gram-negative bacteria play a dominant role in clinics and drug resistance of isolated pathogenic bacteria is a serious problem.Monitoring the trends of pathogenic bacteria′s distribution and drug resistance is very important in guiding the clinical administration of drugs and we should pay attention to fungal infection.

The expressions of decoy receptors of TRAIL in carcinoma of endometrium / 医学研究生学报

Objectives:To determine the expressions of decoy receptors (DcR1 and DcR2) of TRAIL in carcinoma of endometrium. Methods:The expressions of DcR1 and DcR2 in endometrium tissues from 13 carcinoma of endometrium and 7 normal endometrium were detected by immunohistochemical staining.Results: The expressions of DcR1 and DcR2 in carcinoma of endometrium were much lower than in normal endometrium. Conclusions:The decreasing of DcR1 and DcR2 in carcinoma of endometrium may be concerned with its pathogenesis, which may be related to the prevention of endometrium from carcinomatous change.