BACKGROUND: In recent years, ultrasound microbubble gene transfer system has been applied for gene transfection in many parts of the body, but it has been seldom reported to be used for gene transfection in bone parts. OBJECTIVE: To investigate the efficiency and feasibility of ultrasound-targeted microbubble destruction applied for transfection of enhanced green fluorescent protein plasmid into the femoral head of rabbits.METHODS: Japanese big-ear rabbits were randomly divided into five groups: bare transfection, pre-irradiation + bare transfection, ultrasound transfection, pre-irradiation+ultrasound transfection, and repeatable transfection. In the first two groups, ultrasound-targeted gene transfection and irradiation was not used, but in the latter three groups, ultrasound-targeted microbubble destruction was used to transfect enhanced green fluorescent protein (EGFP) plasmid into the femoral head of rabbits. At 1 week after transfection, EGFP expression in femoral head was observed under the fluorescence microscope. RESULTS AND CONCLUSION: EGFP expression appeared in the ultrasound transfection, pre-irradiation + ultrasound transfection and repeatable transfection. The transfection efficiency of EGFP plasmid was significantly higher in the repeatable transfection group than in the other groups (P < 0.01). Obvious injury loci were not observed in the soft tissue and bone tissue slices of ultrasonic irradiation parts in the ultrasound transfection, pre-irradiation + ultrasound transfection and repeatable transfection groups. These results confirm that ultrasound-targeted microbubble destruction is a safe and effective method to transfect EGFP plasmid into the femoral head of rabbits.

A hospital big data-based innovative diagnosis and treatment decision-making support model ( Info Button) was proposed for grass-root medical institutions in Sichuan Province in view of uneven distribution of its medical resources and of beingdifficult and expensive to see a doctoraccording to an analysis of the major health information projects and health information management.How to construct the model was elaborated with its problems pointed out.

Objective To investigate the protective effect of melatonin and its possible mechanism for repairing in the immature white matter damage due to brain hypoxia-ischemia (HI).Methods Forty-eight three-day SD rats after birth were randomly divided into 3 groups:sham-operated(SHAM) group,HI group and melatonin treatment(MT) group.Periventricular white matter damage (PWMD) to animal models were estabished according to Rice modeling.MT group was treated with melatonin pre-operatively,immediately postoperation,1 hour postoperation and 24 hours postoperation via intraperitoneal injection,and the other groups were injected with the same volume of dissolvent.The rats were executed by decollation after 2 days and 14 days.The histological changes in periventricular white matter were observed by HE staining and immunohistochemistry.Results For the 3 groups,the structure in ope-ration side of the white matter in the peripheral ventricles of the brain 2 days postoperation were significant different (P ＜0.05).The O4 positive cells decreased one by one/greatest in the SHAM group[(75.548 ± 7.333)/hpf] followed by MT group [(59.971 ± 3.635)/hpf],and HI group [(40.511 ± 2.848)/hpf] (P ＜ 0.05).The expression of Casepase-3 increased in the SHAM group (107.724 ± 10.266),MT group (132.289 ± 8.537),and HI group (202.168 ± 14.367),and the difference was statically significant (P ＜ 0.05).Ventricular index was greater in operation side of the white matter in the peripheral ventricles of the 14-day-brain in the SHAM group(0.928 ±0.063),MT group (1.813 ± 0.110),HI group (2.752 ± 0.201),increasingly,while absorbance value of myelin basic protein decreased one by one in sequence(39.504 ± 1.673,21.729 ± 1.614,11.344 ± 1.118).Conclusions MT plays a role in protecting the periventricular white matter via inhibiting the apoptosis of oligodendrocyte progenitor cell,and thus benefits the PWMD.

Objective This study was to understand the structure characteristics of prophages in the genome of Enterococcus hirae R17,and also to analyze their interaction relationships with the host bacterium.Methods The gene distribution and gene encoding characteristics of prophages in the genome of Enterococcus hirae R17 were identified using the PHAST software.The virulence gene,antimicrobial resistance genes,and environmental resistance genes in the prophages were also analyzed.Results Three prophages were found on the chromosome of Enterococcus hirae,including two incomplete prophage elements (Prophage-1 and Prophage-2) and one complete prophage (Prophage-3).Some function genes of bacteria were found in the sequence of three prophages,including nucleotide transportation and metabolism related genes.One incomplete prophage carrying erythromycin-and bacitracin-resistance genes was identified in the plasmid,which suggested that prophage induced gene horizontal transfer caused erythromycin-and bacitracin-resistance of Enterococcus hirae R17.Conclusion This study laid a solid foundation for the diversity analysis of prophages of Enterococcus hirae.Prophages played an important role in promotion of antimicrobial resistance of enterococci.Scientists should pay more attention to the spread of antimicrobial resistance and pathogenicity induced by prophages.

AIM: To investigate the changes of expression of SR-BI in phorbol 12-myristate 13-acetate(PMA) differentiated U937 cells.METHODS: U937 cells were cultured with 100 nmol/L PMA in order to differentiate the cells to macrophages.Immunocytochemical method,Western blotting analysis and reverse transcription polymerase chain reaction(RT-PCR) were used to detect SR-BI protein and mRNA during differentiation.RESULTS: Immunocytochemistry showed that after exposure of U937 cells to PMA for 24,48,72 hours,the values of SR-BI protein expression in U937 cells were 15.94?3.56,27.86?4.39 and 9.08?2.37,with the first two higher than that in undifferentiated cells(7.76?1.74,P0.05) increment in the expression of SR-BI protein compared with U937 monocytes.RT-PCR showed that relative SR-BI mRNA expression in different group was 0.112?0.006,0.235?0.014,0.344?0.140 and 0.138?0.010,respectively.CONCLUSION: SR-BI protein and mRNA were increased after differentiation,reached a peak at 48 hours,and decreased at 72 hours.High expression levels of SR-BI in U937 macrophages following PMA differentiation may be correlated with foam cell formation.

Objective To explore the difference of gene expression profiling between normal basilar arteries and basilar arteries of cerebral vasospasm (CVS) after subarachnoid hemorrhage (SAH) in rabbits. Methods cDNA chip of normal basilar arteries and basilar arteries of CVS after SAH in rabbits were downloaded from GEO database. The chip was analyzed and screened by Bioconductor software, and function enrichment and pathway analysis of the differentially expressed genes were analyzed by Cytoscape software. Then 6 adult male Japanese rabbits were used, and randomly divided into normal control group (n=3) and SAH model group (n=3). Rabbit SAH models were established by cisterna secondary-blood-injection method. RNA data of normal basilar artery specimens on the 0 day and basilar artery specimens after SAH on the 5-day were used to validate the parts of differentially expressed genes by qRT-PCR. Results A total of 4356 differentially expressed genes were found in normal basilar arteries and basilar arteries of CVS after SAH in rabbits. Among them, 920 genes were considered to be significant with P-value<0.05, such as GRIK1, MYH13, ZNF45, SAA3, RLN1, MSR1 and others. Function enrichment analysis indicated that the differentially expressed genes were involved in regulation of Ca2+transmembrane transporter activity, negative regulation of ion transmembrane transport, regulation of potassium ion transport, positive regulation of JAK-STAT signaling cascades and other biological processes. Pathway analysis showed that calcium signaling pathway, cGMP-PKG signaling pathway, HIF-1 signaling pathway, PI3K-Akt signaling pathway and other signaling pathways maybe related with the differentially expressed genes. qRT-PCR verification showed that the expression of MSR1 in SAH model group was consistent with that of the chip result. Conclusion The gene expressions of basilar arteries of CVS after SAH in rabbits are significantly different, and MSR1 gene can be used as a potential target for studying the pathological mechanism of CVS.

Objective To observed the influence of preoperative enteral nutrition(EN) on postoperative nutritional status,immune function and complications in elderly patients with colorectal cancer complicating nutritional risk.Methods The NRS2002 nutritional risk screening criteria was used to select 70 elderly patients with colorectal cancer complicating nutritional risk,including 36 cases in the EN group and 34 cases in the control group.The EN support was given in the ENN group on preoperative 3 d.The levels of plasma total protein,prealbumin,albumin,transferrin,total lymphocyte count,plasma D-lactate(D-LAC) and plasma diamine oxidase (DAO) were detected on postoperative 1,3,5 7 d.The intraoperative intestinal cleanliness and postoperative complications were observed.Results The levels of plasma total protein,prealbumin,albumin,transferrin and total lymphocyte count in the EN group were significantly higher than those in the control group and the levels of D-LAC and DAO,and the incidence rates of abdominal infection and wound infection were significantly lower than those in the control group,the differences were statistically significant(P＜0.05).There was no statistically significant differences in the incidence rates of intestinal cleanliness and anastomotic leakage between the two groups (P＞0.05).Conclusion Preoperative EN support therapy in the patients with colorectal cancer complicating nutritional risk can significantly improve clinical prognosis.

Objective:To investigate the predictive value of cerebroplacental ratio (CPR) for adverse perinatal outcomes of induction of labor in prolonged pregnancy.Methods:This retrospective study recruited 315 singleton pregnant women who had induced labor due to prolonged pregnancy (≥41 gestational weeks) in the First Affiliated Hospital of Chongqing Medical University from January 1, 2019 to April 30, 2020. Based on the occurrence of adverse perinatal outcomes (emergency delivery due to persistent abnormal fetal heart rate monitoring, umbilical artery blood pH at birth <7.2, 5 min Apgar scores<7, transferring to neonatal intensive care unit after birth, chorioamnionitis and vaginal delivery converted to cesarean section), they were divided into two groups: case group ( n=76) and normal group ( n=239). Clinical features and umbilical artery blood flow, middle cerebral artery (MCA) flow and CPR measured in the last ultrasound scan before induction were compared between the two groups using student's t-test, Mann-Whitney U test and Chi-square test. Receiver operating characteristic (ROC) curve was used to analyze the predictive values of umbilical artery blood flow, MCA flow and CPR for the adverse perinatal outcomes. Multivariate logistic regression analysis was used to screen the meaningful predictors. Results:Compared with the normal group, the umbilical artery pulsatility index (PI) (0.9±0.1 vs 0.8±0.1, t=-5.458, P<0.001) and the percentage of abnormal CPR (<1.0) increased significantly [21.1%(16/76) vs 6.3%(15/239), χ2=14.190, P<0.001] in the case group, while the MCA-PI and CPR decreased significantly (1.1±0.2 vs 1.3±0.3, t=5.658, P<0.001; 1.2±0.3 vs 1.6±0.5, t=8.940, P<0.001). The areas under the ROC curves of umbilical artery PI, MCA-PI and CPR for predicting adverse perinatal outcomes were 0.71, 0.71 and 0.77, respectively. CPR had the highest sensitivity (0.74) compared with umbilical artery PI (0.68) and MCA-PI (0.71), but the specificity of them were similar (0.67, 0.66 and 0.66). Multivariate logistic regression analysis showed that only CPR was the independent risk factor for adverse perinatal outcomes ( OR=0.028, 95% CI: 0.010-0.080, P<0.001). Conclusions:As an indicator for early prediction of adverse perinatal outcomes of induction of labor in prolonged pregnancy, CPR was more sensitive but less specific.

Background:The pathogenesis of multiple sclerosis (MS) is mediated primarily by T cells,but most studies of MS and its animal model,experimental autoimmune encephalomyelitis (EAE),have focused on CD4+ T cells.The aims of the current study were to determine the pathological interrelationship between CD4 and CD8 autoreactive T cells in MS/EAE.Methods:Female C57BL/6 mice (n=20) were induced by myelin oligodendrocyte glycoprotein (MOG)35-55 peptide.At 14 days after immunization,T cells were isolated from the spleen and purified as CD4+ and CD8+ T cells by using CD4 and CD8 isolation kits,and then the purity was determined by flow cytometric analysis.These cells were stimulated by MOG35-55 peptide and applied to proliferation assays.The interferon-gamma (IFN-γ) and interleukin (IL)-4 secretion of supernatant of cultured CD4+ and CD8+ T cells were measured by enzyme-linked immunosorbent assays (ELISA).For adoptive transfer,recipient mice were injected with MOG35-55-specific CD8+ or CD4+ T cells.EAE clinical course was measured by EAE score at 0-5 scale and spinal cord was examined by staining with hematoxylin and eosin and Luxol fast blue staining.Results:CD8+CD3+ and CD4+CD3+ cells were 86％ and 94％ pure of total CD3+ cells after CD8/CD4 bead enrichment,respectively.These cells were stimulated by MOG35-55 peptide and applied to proliferation assays.Although the CD8+ T cells had a generally lower response to MOG35-55 than CD4+ T cells,the response of CD8+ T cells was not always dependent on CD4.CD8+ T cell secreted less IFN-γ and IL-4 compared with CD4+ T cells.EAE was induced in wildtype B6 na(i)ve mice by adoptive transfer of MOG35-55-specific T cells from B6 active-induced EAE (aEAE) mice.A similar EAE score and slight inflammation and demyelination were found in naive B6 mice after transferring of CD8+ T cells from immunized B6 mice compared with transfer of CD4+ T cells.Conclusion:Our data suggest that CD8+ autoreactive T cells in EAE have a lower encephalitogenic function but are unique and independent on pathogenic of EAE rather than their CD4+ counterparts.

OBJECTIVETo compare the detection sensitivity of epidermal growth factor receptor (EGFR) mutations between allele specific oligonucleotide PCR (ASO-PCR) and bi-loop probe and specific primer quantitative PCR (BPSP-qPCR).

METHODSA total of 96 non-small cell lung cancer specimens were selected from West China Hospital from September 2009 to December 2010. ASO-PCR was developed to detect the presence of classical EGFR mutations. A total 39 available specimens were also tested by BPSP-qPCR.

RESULTSEGFR mutation detection rate was 30.2% (26/96) by ASO-PCR. The mutation rate was higher in female than in male patients [45.5% (20/44) vs. 17.3% (9/52), P = 0.003], non-smokers than smokers [44.1% (26/59) vs. 8.1% (3/37), P < 0.001] and adenocarcinomas than other subtypes of lung cancer [37.0% (27/73) vs. 8.7% (2/23), P = 0.01]. Among mutation negative cases by ASO-PCR, BPSP-qPCR increased the rate of detection of 19-del and L858R mutation by 10.3% (4/39) in adenocarcinomas and non-smoking subset. Overall, the mutation detection rate of BPSP-qPCR was higher than that of ASO-PCR [66.7% (26/39) vs. 41.0% (16/39), P = 0.02].

CONCLUSIONBPSP-qPCR has a better detection sensitivity than that of ASO-PCR.

Adenocarcinoma ; genetics ; Carcinoma, Non-Small-Cell Lung ; genetics ; DNA Mutational Analysis ; Female ; Genes, erbB-1 ; Humans ; Lung Neoplasms ; genetics ; Male ; Middle Aged ; Mutation ; Polymerase Chain Reaction ; methods ; Receptor, Epidermal Growth Factor ; genetics ; Sensitivity and Specificity ; Sex Factors ; Smoking