Objective To investigate the adhesion mechanism of Lactobacillus acidophilus FN001外to Peyer's patches. Methods Adhesion of L. acidophilus FN001 to mice Peyer's patches was studied in vitro using a fluorescent quantization method. The nature of adhesion mediator was studied by the effects of physical, chemical and enzymatic pre-treatments of the bacteria on their adhesion and effect of sugars on in- hibition of adhesion. The presence of lectin-like proteins in the cell surface was determined by hemagglutina- tion. Effect of L. acidophilus FN001 on inhibition of adhesion of pathogens to Peyer's patches was also stud- ied. Results The adhesion of L. acidophilus FN001 was strongly inhibited in the presence of D-mannose and methyl-ct-D-mannoside. Pretreatment of L. acidophilus FN001 with pepsin and trypsin decreased the ad- hesive capacity indicating that cell surface proteins are involved in adhesion to Peyer's patches. L. acidophi- lus FN001 could agglutinate rabbit red cell in mannose specific manner and protease pretreatment could de-crease hemagglutinin, suggesting that L. acidophilus FN001 has mannose specific lectin (s). In adherence inhibition assay, L. acidophilus NF001 could significantly inhibit adhesion of E. coli ATCC25922 to Peyer's patches when L. acidophilus NF001 were applied to Peyer's patches first or at the same time with pathogen. Conclusion It was concluded that a mannose-specific protein mediated adhesion of L. acidophilus FN001 to the Peyer's patches, and L. acidophilus FN001 could inhibit adhesion of pathogen with similar lectins speci- ficity to Peyer's patches.

Objective To explore extracellular signal-regulated kinase ( ERK1/2 ) expression in the role of curcumin inhibited staurosporine (STS)-mediated neurons toxic injury through added PD098059, and to clarify ERK1/2 mediated inhibitory role of curcumin on STS-induced neurons toxic injury.Methods The neurons toxic injury model of primary cultured hippocampal neurons was established by STS.The experiment was divided into six groups:normal control group, STS model group, PD098059 +STS model group, curcumin +STS pretreatment group, curcumin+PD098059+STS treatment group and curcumin treatment group.The cell viability were determined by MTT method, lactate dehydrogenase (LDH) release rate, cell toxicity were detected, nuclear shape were observed by DAPI staining, and ERK1/2 expression were detected by Western blot method.Results The cell viability of curcumin +STS pretreatment group was significantly higher than STS model group ( P <0.001 ); the cell viability had no significant difference between PD098059 +STS model group and curcumin +PD098059 +STS treatment group;compared with curcumin +STS model group , the cell viability of curcumin +PD098059 +STS treatment group was significantly decreased ( P<0.001 ).LDH results showed that the nerve cell toxicity of curcumin +STS pretreatment group was obviously less than STS model group (P<0.001).The cell nuclear shape showed typical apoptosis morphological characteristics in STS model group, and curcumin inhibited the effect of STS mediated-neuronal apoptosis.ERK1/2 protein expression in curcumin +STS pretreatment group significantly increased compared with STS model group ( P <0.001 ) .Conclusion Curcumin inhibited STS-mediated neurons toxicity injury by up-regulating ERK1/2 expression.After PD098059 blocking the nerve cells ERK1/2 synthesis, the inhibitory action of curcumin failed to implemented, which illustrated that ERK1/2 mediated curcumin to inhibit STS-induced neuronal toxic injury.

Objective: To investigate the level and timeliness of CD4+T lymphocyte cell (CD4 cell) counts at first measurement among newly reported HIV/AIDS cases in Taizhou City, Zhejiang Province from 2012 to 2021, so as to provide insights into improving the management of HIV/AIDS cases. Methods: Demographic data and first measurement of CD4 cell counts of newly reported HIV/AIDS cases in Taizhou City from 2012 to 2021 were collected from the HIV/AIDS Comprehensive Control System of Chinese Disease Prevention and Control Information System. The level and time of CD4 cell counts at first measurement were descriptively analyzed, and factors affecting CD4 cell counts at first measurement were identified using a multivariable logistic regression model. Results: A total of 4 834 newly reported HIV/AIDS cases were recorded in Taizhou City from 2012 to 2021, including 3 889 men (80.45%), 2 005 cases at ages of 20 to 39 years (41.48%), and 2 130 farmers (44.06%). There were 1 664 cases (34.42%) with CD4 cell counts of <200/mm3 at first detection, 2 576 (53.29%) with CD4 cell counts of 200/mm3 to 499/mm3, and 594 (12.29%) with CD4 cell counts of ≥500/mm3. The proportion of CD4 cell counts of <200/mm3 showed a tendency towards a rise from 2012 to 2021 (χ2trend =4.250, P<0.001). There were 3 465 cases (71.68%) that had an interval of ≤14 days between the first detection of CD4 cell counts and confirmatory HIV test, 740 (15.31%) that had an interval of >14 to 30 days, 315 (6.52%) that had an interval of >30 to 90 days, 135 (2.79%) that had an interval of >90 to 180 days, and 179 (3.70%) that had an interval of >180 days. The proportion of an interval of ≤14 days appeared a tendency towards a rise from 2012 to 2021 (χ2trend=6.874, P<0.001). Multivariable logistic regression analysis identified women (OR=0.630, 95%CI: 0.529-0.751), age of ≥20 years (OR: 1.958 to 3.218, 95%CI: 1.216-5.412), other or unknown routes of HIV infection (OR=1.785, 95%CI: 1.100-2.896), and identification by medical institutions (OR=1.779, 95%CI: 1.497-2.114) as factors affecting CD4 cell counts of <200/mm3 at first measurement. Conclusions The timely detection of CD4 cell counts at first measurement gradually increased with year among newly reported HIV/AIDS cases in Taizhou City from 2012 to 2021; however, there were still 34.42% of cases with CD4 cell counts of <200/mm3. Gender, age, route of HIV infection, and sample source were factors affecting CD4 cell counts of <200/mm3 at first measurement.

This article described the background,concept,characteristic and objective of the research-based nursing,systematically introducing the main measures including management mechanism, nursing service,nursing staff training,and nursing scientific development.Other areas covered include innovation management mechanism,updating service philosophy,improving nursing staff training,and constructing scientific research platform.

Objective To investigate the feasibility of macrophages as cell carriers to deliver floate modified oxygen loaded ultrasound contrast agent . Methods The phagocytic activity of macrophages was analyzed by ink phagocytose test , and the expression of folate recepters ( FRs ) on macrophages cell membrane surface was tested by immunofluofluorescence assay . Oxygen/paclitaxel loaded lipid microbubbles( OPLMB) and folate‐targeted OPLMB ( TOPLMB) were synthesized by mechanical shock method and incubated with macrophages in vitro . According to different treatment conditions ,the cells were divided into three groups:group A ( OPLMB) ,group B ( free folic acid + TOPLMB) and group C ( TOPLMB) , the fluorescence intensity of the cells were observed under fluorescence microscope ,and the phagocytic percentage and the phagocytic index of macrophages uptake OPLMB and TOPLMB were observed by bright field microscope . Results The phagocytic percentage of macrophages phagocytose ink was (99 .3 ± 1 .0)% ,FRs was highly expressed on macrophages cultured in vitro . After incubation for 30 minutes ,the fluorescence intensity of group C was significantly higher than those of A and B ,the phagocytic percentage in three groups were (19 .5 ± 0 .2)% ,(21 .0 ± 0 .2)% and (81 .2 ± 10 .0)% respetively . The phagocytic percentage of group C were significantly higher than those in group A and group B ( P <0 .05) . Conclusions Macrophages cultured in vitro possess highly phagocytic activity and these cells highly express FRs ,and can be used as cell carriers to deliver floate modified oxygen loaded multimodality ultrasound contrast agent .

Objective To explore the survival/proliferation,apoptotic and death effects of keratinocyte growth factor (KGF) in alveolar epithelial type Ⅱ cells (AT Ⅱ Cs) exposed to hyperoxia.Methods Primary culture of AT Ⅱ Cs from the Sprague-Dawley rat fetuses was studied under room air condition (210 mL/L O2) and hyperoxic condition (950 mL/L O2) for 0.5-12.0 h.Various concentrations of KGF (15 μg/L,25 μg/L,50 μg/L,75 μg/L,100 μg/L)were added into the cell cultures.Cells were randomly divided into room-air group,room-air-KGF group,hyperoxic-exposure group and hyperoxic-exposure-KGF group.The levels of intracellular reactive oxygen species (ROS),cleaved cysteinyl aspartate specific proteinase-3 (Caspase-3),cell death and proliferation of AT Ⅱ Cs were measured by flow cytometer,Western Blot,release of lactate dehydrogenase assays (LDH assays) and 3-(4,5-Dimethyhhiazol-2-yl)-2,5-diphenyhetrazolium bromide assays (MTT assays),respectively.Results Under room air condition,KGF could significantly increase AT Ⅱ Cs proliferation with 15-100 μg/L in a dose-dependent manner and significantly decrease LDH production at concentrations of 25-100 μg/L.Exposure to hyperoxia resulted in a significant increase in intracellular ROS production in AT Ⅱ Cs in a time-dependent manner compared with that of the room air group.Cell viability decreased and LDH release increased significantly in a time-dependent manner when AT Ⅱ Cs were exposed to 950 mL/L O2 for more than 4 h.After exposure to hyperoxia for 0.5 h and 1 h,KGF could significantly increase AT Ⅱ Cs proliferation in 15-75 μ g/L and significantly decrease LDH production at concentrations of 25-75 μg/L.After exposure to hyperoxia up to 4 h,higher viability was observed in 15 μg/L and 25 μg/L KGF group,and lower death rate presented in 25-100 μg/L KGF group.Further,prolnged hyperoxic exposure for 8 h,high viabilitv was shown only in 50 μg/L KGF group,and less death rate was observed only in 75 μg/L KGF group.In addition,no significant difference in viability and mortality was found between hyperoxic group and hyperoxic-KGF group after hyperoxic exposure for 12 h.Expression of cleaved Caspase-3 was significant higher after 4 h and 8 h hyperoxic exposure than that in room-air group ;at the same time,by adding 25 μg/L and 75.μg/L KGF led to decreased expression of Caspase-3 was detected,compared to hyperoxic group.Conclusions KGF may promote survival/proliferation,inhibited apoptosis and death of rat fetal AT Ⅱ Cs in room air condition or under temporary exposure to hyperoxia in vitro.However,prolonged exposure to hyperoxia may decrease the sensitivity of AEC Ⅱ Cs to KGF and limit its protective effects on lung injury.

Objective To detect the levels of 25-hydroxyvitamin D3 and analyse the related clinical factors in patients with chronic kidney disease (CKD).Methods Patients with CKD in our department from March 1st to July 1st were enrolled continuously.The level of 25-hydroxyvitamin D3 and intact parathyroid hormone(iPTH) were detected by electrochemiluminescence and immunochemiluminescent respectively.Serum calcium,phosphorus and albumin were measured by automatic biochemical instrument.Results 127 patients were selected and the average age was (60.9?15.3).The mean level of 25-hydroxyvitamin D3 was (12.06?6.41)?g/L.82.6% patients had vitamin D deficiency and 96.9% patients had vitamin D insufficiency.The level of 25-hydroxyvitamin had no statistics difference D3 between stage 1,2 and 3 CKD patients but was much hihger than that of stage 4 and non-haemodialysis stage 5 patients.The level of 25-hydroxyvitamin D3 was not related to serum calcium,phosphorus and iPTH,while positively related to albumin and eGFR and negatively related to serum creatinine and total cholesterol.Conclusion Vitamin D deficiency and insufficiency are frequent in CKD patients and deteriorate with the progress of kidney function impairment.The level of 25-hydroxyvitamin D3 is not related to the traditional CKD-MBD index such as serum calcium,phosphorus and iPTH.

In the previous study, a high-throughput screening method was established to find the antagonists of CD36. In the present study, a new compound named IMB-1680 was found using this method. The anti-atherosclerotic activities of IMB-1680 were then evaluated. Dose-dependent activities of IMB-1680 were detected by using Sf9 [hCD36] and CHO [hCD36] models. Fluorescence microscopic photography and flow cytometry were used to analyze uptake of mLDL. Foam cell test with RAW264.7 macrophages was used to examine lipid accumulation. The results showed that IMB-1680 inhibited CD36 activity with IC50 of 2.80 and 8.79 micromol x L(-1) in Sf9[hCD36] and CHO [hCD36] cells, respectively. Fluorescence microscopic photography and flow cytometry revealed that IMB-1680 could significantly reduce DiI-AcLDL uptake. Meanwhile, IMB-1680 also could reduce lipids accumulation in RAW264.7 macrophages. In all, the data indicated that IMB-1680 might be a potent effective anti-atherosclerotic leading compound.

Objective To explore the change of CD4+CD25+Foxp3+ regulatory T (Treg) cells during aging and the relation with lung tumor. Methods The Lewis lung cancer model was set up in C57BL/6 female mice, and the 36 mice were divided into young health group, middle-aged health group, elderly health group, young tumor group, middle-aged tumor group and elderly tumor group. The percentages of CD4+CD25+Foxp3+ Treg in CD4+ T cells in mice spleen cells were measured by flow cytometry, for reflecting the quantity of CD4+CD25+Foxp3+ Treg cells. And the level of Foxp3 mRNA in splenocyte was tested by real-time PCR method. Results The level of CD4+CD25+Foxp3+/CD4+ T cells and the quantity of Foxp3 mRNA were higher in tumor groups than in healthy groups(both P＜0.05 ). Besides, in the healthy groups, there were statistical differences in the level of CD4+CD25+Foxp3+/CD4+ T cells (F=47.70, P=0.000) and the quantity of Foxp3 mRNA among the different months groups. Accumulation of the CD4+CD25+Foxp3+ Treg cells was accompanied with aging, the elderly mice contained a significantly larger population of CD4+CD25+Foxp3+ Treg cells in their spleen when compared with the younger counterparts, and the highest was the elderly tumor group. So it was with the functional gene Foxp3 mRNA (F=6.56, P=0.009). Conclusions The results suggest a close relationship of the change of CD4+CD25+Foxp3+Treg cells with aging and the genesis and development of lung tumor.

Objective To explore whether the change of S phase kinase-associated protein 2 (Skp2) expression could regulate mesangial cell proliferation. Methods Skp2 siRNA and control siRNA, pIRES-GFP-Skp2 plasmid and pIRES-GFP plasmid were designed and synthesized. Cell transfection was performed by Lipofectamine 2000. Skp2 mRNA and protein levels were detected by semiquantitative PCR and Western blotting respectively. Primary culture rat mesangial cells were divided into 6 groups: 0%FCS, 20%FCS, 10%FCS+pIRES-GFP plasmid, 10%FCS+pIRES-GFP-Skp2 plasmid, 20%FCS+control siRNA, 20%FCS+Skp2 siRNA. Cell number was detected by MTT. S phase entry was measured by BrdU labeling. Cell cycle profile was determined by flow cytometric analysis. Results Skp2 mRNA level was significantly down-regulated by Skp2 siRNA compared to control siRNA. Skp2 protein level increased after pIRES-GFP-Skp2plasmid transfection compared to pIRES-GFP plasmid. MTT, BrdU labeling and cell cycle profile demonstrated that cell number (A: 0.419±0.088 vs 0.305±0.036, P＜0.01) and S-phase cells (BrdU labeling positive cell: 0.21±0.04 vs 0.15±0.03, P＜0.01;S-phase cell number:20.18±0.64vs 14.33±0.37, P＜0.01) obviously increased after Skp2 plasmid transfection, while decreased after Skp2 siRNA transfection compared to control groups (A: 0.328±0.069 vs 0.482±0.133, P＜0.01;BrdU labeling positive cell: 0.17±0.01 vs 0.24±0.00, P＜0.01; S-phase cell number: 16.52±0.75vs 23.81 ±1.25, P＜0.01). Conclusion Over-expression of Skp2 stimulates mesangial cell proliferation while down-regulated expression of Skp2 inhibits mesangial cell proliferation.