0bjective To study the expression of Fas in hippocampus of cerebral ischemia-reperfusion rats after treatment with monosialoganglioside (GM1). Methods Seventy-two SD rats were randomly divided into sham group (n=8), GM1 treatment group (n=32) and ischemia-reperfusion (I/R) group (n=32). According to the different time points (6 h, 12 h, 24 h and 3 d) of I/R, there were four subgroups in GM1 and I/R groups respectively. The expression of Fas in hippocampus was examined by immunochemistry and Western blot methods in all groups. Results Results of immunochemistry method showed that there was a little expression of Fas (the mean optical density was 0.17±0.02) in hippocampus of sham group, and the positive expression of Fas at different time points were increased significantly after reperfusion. The mean values of opti-cal density at different time points were 0.42±0.11, 0.51±0.13, 0.55±0.13 and 0.62±0.15 in I/R group, which reached to the peak at 3 d . The positive expression of Fas at different time points were significantly decreased in GM1 group (0.29 ± 0.09, 0.34±0.11, 0.37±0.12 and 0.43±0.12) than that of I/R group (P＜0.05). Conclusion Monosialoganglioside participated in the pathogenesis of cerebral ischemia-reperfusion injury by down-regulating the expression of Fas.

OBJECTIVE: To research the prevalences of four kinds of bacteria including Alloiococcus otitidis, Streptococcus pneumonia, Haemophilus influenzae, and Moraxella catarrhalis in children with chronic otitis media with effusion (SOM) of the middle ear effusion, and the reproduction of the nasopharynx, so as to explore their meaning for the children with SOM. METHOD: Alloiococcus otitidis, Streptococcus pneumonia, Haemophilus influenza, and Moraxella catarrhal were investigated in the samples obtained from middle ear effusion and nasopharyn- geal swabs, using PCR and conventional bacterial culture methods. RESULT: By bacterial culture, the pathogen detection rate from middle ear effusion was 3.6%,while the nasopharynx was 54.0%, the detection rate of Streptococcus pneumonia, Haemophilus influenza, Moraxella catarrhalis was 10.8%, 27.0%, 4.5%, respectively, the drug susceptibility results for 51 samples of bacterial culture positive showed that 39 cases was sensitivite to the β-lactam antibiotic; By PCR, the number of detecting various kinds of bacteria simultaneously in middle ear effusion or in the nasopharynx were 6 and 34. The bacteria prevalences of S. pneumoniae, H. influenzae, M. catarrhalis, and A. otitidis are 5.4%, 5.4%, 3.6%, and 42.3% in the middle ear effusion, are 25.2%, 27.0%,13.5% and 34.2% in nasopharyngeal, respectively. CONCLUSION (1) PCR method is more sensitively detecting the bacteria than conventional bacterial culture methods. (2) The chronic SOM of children may be a combination of mixed bacterial infection, A. otitidis may be the most common pathogen of children SOM. (3) For children of SOM, if antibiotics are chosen to be used early in the disease, we suggest using the β-lactam antibiotics.
Bacteria ; Bacterial Infections ; complications ; Child ; Haemophilus influenzae ; isolation & purification ; Humans ; Moraxella (Branhamella) catarrhalis ; isolation & purification ; Nasopharynx ; Otitis Media with Effusion ; complications ; microbiology ; Polymerase Chain Reaction ; Prevalence ; Streptococcus pneumoniae ; isolation & purification

Objective To explore the feasibility of labeling in vitro rabbit osteoblasts with Qtracker and the features of Qtracker-labeled rabbit osteoblasts. Methods A healthy male rabbit, 3 months old, weighing 2 kg, was used in this study. After bone marrow was aspirated, bone marrow stromal cells (BMSCs) were isolated and cultured using the adherence method in vitro. The third passage of BMSCs was induced into osteablasts before incubation with Qtracker at concentrations of 1, 2, 4, 8, 16, 32 nmol/10~6 cells (Groups A, B, C, D, E, F respectively). Cells not labeled by Qtracker served as negative control (Group G). The following parameters were measured: induction, differentiation and determination of rabbit osteoblasts; the optimal mass concentration of Qtracker labeling by fluorescence microscopy and flow cytometry; the cell sur-vival rates at various concentrations of Qtraeker labeling by trypan-blue exclusion; Qtracker-labeled cell pro-liferation by MTr. Results The primary and the passage rabbit BMSCs were chiefly of fusiform shape. Rabbit BMSCs differentiated into osteoblasts following induction. The osteoblasts cytoplasm showed green fluorescence under fluorescence microscopy after being labeled by Qtracker. The mean labeling rate increased with the increased concentration of Qtracker, reaching up to (93.58±2.08) % after incubation at 8 nmol/ 10~6 cells by fluorescence microscopy, and (95.24±1.31) % by flow cytometry. There were no significant differences between Groups D, E, F(P>0.05), but significant differences were found between Groups A, B, C and Groups D, E, F (P<0.05). The labeling rate for Group G was 0. The cell survival rates were all above 96% (P>0.05) . No significant differences were found in the cell proliferation among various con-centrations (P>0.05). Conclusions Qtraeker can be used as a labeling marker for rabbit osteoblasts. When the concentration is at 8 nmol/10~6 cells, optimal labeling effect can be achieved. Rabbit osteoblasts labeled with Qtracker are of high efficiency and safety.

Objective To explore the proliferation and differentiation of osteoblasts from 2 sources co-cultured with SD rat Schwann cells(SCs) . Methods Bone marrow stromal cells (BMSCs) were obtained by washing the femoral and tibial bone marrow cavities in SD rats. Osteoblast differentiation of the third passage of BMSCs was induced by incubation in osteogenic medium. Primary rat calvarial osteoblasts were obtained by digestion of the calvarial bone in one day old SD rats. The cells were cultured in DMEM supplemented with 10% fetal bovine serum(FBS) . SCs of passage 2 were obtained by digestion of sciatic nerve. The SCs were identified by S-100. The proliferation of 2 kinds of osteoblasts co-cultured with SCs was tested using 96 co-culture plate by methyl thiazdyl tetrazolium(MTF). Real-time PCR was used to test the osteoblast differentiation through co-culturing with SCs in 3 d and 7 d. The osteoblasts were implanted in the subtus chamber. The SCs were implanted in the superior chamber. Results SCs enhanced significantly the proliferation of calvarial osteoblasts at 7 time points. The expression levels of OPN mRNA, OCN mRNA, ALP mRNA, and BMP-2 mRNA of the osteoblasts were significantly lower in the experiment group than in the control group in 3 d and 7 d. SCs also enhanced significantly the proliferation of the induced osteoblasts in 5 d, 7 d and 9 d. The expression levels of OPN mRNA, OCN mRNA, ALP mRNA, and BMP-2 mRNA of the induced osteoblasts were significantly higher in the experiment group than in the control group in 3 d and 7 d, except the level of ALP mRNA in 7 d.Conclusions The BMSCs-induced osteoblasts cocultured with SCs may be used as seed cells to construct neurotized tissue engineered bone.

Objective:To systematically review parents ′ experience of caring for children with chronic kidney disease (CKD) to fully understand care needs and improve the psychological state and caregiving quality of parents. Methods:The qualitative studies on parents ′ experience of caring for children with CKD were retrieved from following databases, including PubMed, Web of Science, CNKI, VIP, CBM, and WanFang Data from inception to March 2020. The quality of included researches was evaluated according to the JBI Critical Appraisal Tool for qualitative studies in Australia. The results were integrated by pooled integration methods. Results:A total of 14 studies were included. 69 results were summarized and integrated to form 10 categories. These categories extracted 4 integrated results: parents' physical and mental condition are affected, relationship between parents and their support system has changed, reconstruction of parents ′ life, unmet care needs and problems. Conclusions:Parents are crucial to the disease management of children with CKD, so clinical medical staff should not only provide medical services for children, but also pay more attention to the psychological status and needs of parents, so as to provide guidance and support to promote parents to better implement care and disease management for children.

Purpose Recently the incidence of bone and joint degeneration is increased,with the increasing of cardiovascular disease complication.This study aims to utilize realtime three-dimensional echocardiography (RT-3DE) in evaluating left ventricular remodeling index (LVRI),to guide treatment and improve the survival rate.Materials and Methods Fifty-seven cases of healthy volunteers (normal control group) and sixty-seven patients with bone and joint degeneration (research group) were respectively divided into two subgroups according to the age.Both groups underwent conventional echocardiography and RT-3DE.The LVRI,left ventricular end diastolic volume index (LVEDVI),left ventricular mass index (LVMI),left ventricular end systolic volume index (LVESVI) and left ventricular ejection fraction (LVEF) measured by different methods were analyzed,and the differences and correlations between groups and within each group were compared.Results The age of normal control group and research group was not significantly different (P＞0.05).The LVMI and LVRI in research group were significantly higher than those in the normal control group with same age (P＜0.05).The LVEDVI,LVMI and LVRI measured by two-dimensional echocardiography and M-mode echocardiography were higher than those measured by RT-3DE,and the difference was statistically significant (P＜0.05).Bland-Altman agreement analysis showed good agreement of measured LVRI using 2D echocardiography and RT-3DE.Conclusion Patients with bone and joint degeneration have left ventricular remodeling.RT-3DE can accurately assess LVRI with the advantages of convenience and lack of radiation,which can be widely used in clinic.

Objective To investigate the effect of high mobility group box‐1(HMGB1) on the migration of vascular smooth cells (VSMCs) and the role of TLR4‐dependent PI3K/Akt pathway in the process .Methods Primary VSMCs were isolated from the thoracic aorta of male SD rats and cultured in vitro .Control group ,TLR4 siRNA transfected group ,control siRNA transfected group and PI3k inhibitor (LY294002) intervention group were stimulated by HMGB1 (0 .1-1 000 .0 ng/mL) .Expression of TLR4 mRNA was detected by RT‐PCR ,protein expression of TLR4 ,Akt ,pAkt ,PI3K were detected by Western blot .Activity of the im‐munoprecipitated PI3K enzyme was assessed in a competitive ELISA .The migration and cell viability of every groups were ob‐served .Results HMGB1 (0 .1 -1 000 .0 ng/mL) stimulated VSMCs migration in a dose‐dependent manner and incubation of VSMCs with 100 ng/mL caused a rapid migration (P< 0 .05) .At the concentrations used ,HMGB1 did not cause any cytotoxic effects (P<0 .05) .Migration of VSMCs toward HMGB1 was significantly inhibited by silencing of TLR4 (P<0 .05) .Pretreated cells with TLR4 siRNA or the PI3K inhibitor LY294002 could markedly block PI3K/Akt pathway activation and VSMCs migration mediated by HMGB1 (both P<0 .05) .Conclusion HMGB1 stimulated VSMCs migration in a dose‐dependent manner and TLR4‐dependent PI3K/Akt signaling pathway played an important role in the migration of VSMCs mediated by HMGB1 .This research indicates that TLR4‐dependent TLR4/PI3K/Akt signaling pathway could be the target in the treatment of obstructive cardiovascu‐lar disease .

Objective To elucidate the role of renal progenitor-like tubular cells in the repair process after acute tubular necrosis(ATN)induced by ischemia. Methods Rat ATN was developed by clamping left kidney artery for 60 minutes,and bromodeoxyuridine(BrdU),a cell division and proliferation marker,was administrated one hour before rats were sacrificed.Kidneys were isolated at 1,3,5,7,14,21,28 days after injury.The proliferative and apoptotic cells were determined by immunostaining using anti-BrdU,Pax2(an embryonic renal marker),vimentin(an immature mesenchymal cell marker),and activated caspase-3(a cell apoptosis marker). Results Cell death was found in tubules at day 1 after ischemia and reperfusion injury.BrdU-positive cells were dramatically increased and reached peak at day 3 after injury.In addition,the number of BrdU positive cells in the contralateral kidney was significantly increased compared to sham operated group.Double immunostaining showed that BrdU-positive cells co-expressed Pax2 or vimentin,but not activated caspase-3. Conclusions Renal progenitor-like tubular cells may play a predominant role in repair process following ATN in rats.They may dedifferentiate,proliferate,and then redifferentiate into mature tubular cells.Growth factors may regulate the repair process.

Objective Based on active monitoring MRSA carriage for hospitalized patients, the relationship between colonization pressure and MRSA cross transmission in wards without rigorous contactisolation measures was analyzed, and the role of colonization pressure in predicting MRSA cross transmission was further evaluated. Methods From March to December 2009, active MRSA colonization screening was performed for 240 hospitalized patients in emergency ward and 94 cases in RICU in our hospital. rep-PCR method was employed to do homology analysis on MRSA strains obtained in this study. MRSA weekly colonization pressure, threshold colonization pressure ,cross transmission rate were calculated respectively. RR of MRSA cross transmission under higher level of colonization pressure and lower level of colonization pressure was analyzed. Results MRSA carriage rates on admission for patients in emergency wards and RICU were 6. 25% (15/2A0) and 13. 83 % (13/94) ,and MRSA cross transmission occurred in 13 weeks and 14 weeks in above two units, respectively. Threshold colonization pressures for above two units were 6. 49%and 17. 66%, respectively. For emergency ward, the MRSA cross transmission rate under higher level of colonization pressure was significantly higher than that under lower level of colonization pressure (x2 = 7. 10,P＜0. 01), the RR of MRSA transmission was 9. 61 (95% CI:1. 25-74.00). For RICU, the MRSA cross transmission rate under higher level of colonization pressure was significantly higher than that under lower level of colonization pressure(x2 = 12. 60, P＜0. 01 ), the RR of MRSA transmission was 15.87 (95% CI:2. 06-122. 10). Conclusions Higher level of colonization pressure is an important risk factor for MRSA transmission, and average colonization pressure can be used as a prediction index for MRSA transmission and strengthening prevention and control measures.

Objective To investigate the association of hepatitis B virus(HBV) S gene quasispecies with the outcome of HBV infection.Methods Serum samples were collected from three chronic HBV carriers, three chronic hepatitis B and three chronic severe hepatitis B patients.All subjects were male and with HBV genotype C.HBV S gene was amplified, and 20 clones of HBV fragment were randomly selected and sequenced from each sample.SPSS 15.0 software was adopted for analysis.Results Quasispecies complexity of HBV S gene in chronic HBV carriers and chronic hepatitis B tended lower than that of the severe chronic hepatitis B, but the difference was not of statistical significance (P＞0.05).In T cell epitope 45, 47, 85 amino acid sites of the HBV S gene, the constitution of quasispecies in the chronic hepatitis B was more complex than that of the HBV carriers (P=0.01), but compared with the severe chronic hepatitis, the difference was not significant (P=0.06).The computer model showed that both the dominant clones and the non dominant clones could effectively bind to the receptors of cytotoxic T lymphocytes.Conclusion Quasispecies in some T cell epitopes of HBV S gene may be related with the clinical outcome of hepatitis B.