【Objective】 To explore the relationship between the storage time of leukodepleted red blood cells and transfusion adverse reactions by analyzing the occurrence of transfusion adverse reactions of patients after leukodepleted red blood cells transfusion from four hospitals. 【Methods】 By using the electronic medical record management system, the collection and transfusion dates of leukodepleted red blood cells from four hospitals in Enshi Prefecture from 2018 to 2022, as well as the information on transfusion adverse reactions, were retrieved. 【Results】 From 2018 to 2022, a total of 697 61 bags of leukodepleted red blood cells were transfused in four hospitals, resulting in 166 cases of transfusion adverse reactions, among which 93 were allergic reactions, 63 were non hemolytic febrile reactions, and 10 were others, with a total incidence rate of transfusion adverse reactions at 0.24%. The average storage time of leukodepleted red blood cells with and without transfusion adverse reactions was (20.25±6.31) and (19.88±5.50) days, respectively. With a storage time of 7 days as the threshold, the incidence of transfusion adverse reactions was the lowest for a storage time of 15~21 days. The incidence of transfusion adverse reactions of leukodepleted red blood cells in two groups (with storage days ≤21 days and >21 days) was not statistically significant(P>0.05). 【Conclusion】 Allergic reactions were the main type of transfusion adverse reaction caused by leukodepleted red blood cells, and the incidence of transfusion adverse reactions decreased and then increased with the prolongation of the storage time of leukodepleted red blood cells. There was no significant difference in the incidence of transfusion adverse reactions with leukodepleted red blood cells stored for ≤ 21 days and >21 days.

Objective To survey the current status of professional value among health workers in grass-roots medical institutions of Wenzhou city and to analyze the influencing factors.Methods A survey on professional value was conducted from October to December 2017 among health workers from 14 primary medical institutions in 4 urban districts of Wenzhou City selected by stratified random sampling method.The self-designed questionnaires were used for the survey.A total of 350 questionnaires were distributed and 331 were returned,of which 302 were valid.The status and influencing factors of professional value were analyzed.Results The overall score of professional value was (96.66 ± 14.94).The univariate analysis showed that the professional value scores were associated with educational level (F=3.851),income(F=2.660),professional title(F=8.030),position(F=11.658) and specialty (F=4.667) of the health workers(all P＜0.05).The multivariate analysis showed that the gender(male:OR=0.53,95％ CI:-1.077--0.192,P=0.005),educational level (high school and below:OR=3.06,95％ CI:0.007-2.233,P=0.049),income (2 000-4 000 Yuan:OR=0.07,95％ CI:-4.783-0.627,P=0.011),professional title (no title:OR=0.27,95％CI:-2.397--0.232,P=0.017;Junior title:OR=0.31,95％CI:-2.197--0.153),P--0.024;intermediate title:OR=0.23,95％ CI:-2.533--0.366,P--0.009),position (doctor:OR=0.13,95％ CI:-3.235--0.815,P=0.001;nurse:OR=0.20,95％ CI:-2.883--0.368,P=0.011;technician:OR=0.15,95％ CI:-3.076--0.723,P=0.002)were influencing factors of professional value(all P＜0.05).Conclusion The professional values among health workers in Wenzhou grass-roots medical institutions are relatively low.The male gender,low educational level,low income,low professional title or no title and positions are influencing factors of professional value.

Early diagnosis and intervention is an important way to delay the progress of Alzheimer′s disease (AD).Compared with cerebrospinal fluid, blood sampling is not invasive and easy to be obtained in clinic practice.In this study, the serum samples of 9 controls, 10 AD and 12 mild cognitive dysfunction (MCI) patients were analyzed and compared through one by one analysis to screen potential markers for AD diagnosis.The experimental results showed that VGFYESDVMGR of α-2-macroglobulin peptide was closely related to the late stage of AD disease, and the large amount degradation of apolipoprotein C-Ⅲ, histone H1.2 and histone H1.4 was significantly related to early stages of AD progression.The characteristics of serum peptidome were different for the early and late AD, and these four proteins may be used as potential biomarkers of AD disease.In addition, the obvious ladder sequence characteristic was observed for apolipoprotein C-Ⅲ and histone H1, which could partly explain why the peptides distribution in different samples was somewhat contingent.On the contrary, the distribution at protein level was more stable.Finally, it was confirmed that the peptides of proteins such as fibrinogen α-chain, thymosin β-4 and patchy proteins were the dominant peptides in all serum samples.Overall, this study showed that the method of using serum peptidomics to diagnose AD was possible.The results may provide evidence and references for the large-scale clinical validation of AD.

Aim To investigate the protective effects of sphingosine 1-phosphate ( S1 P ) postconditioning on hypoxia/reoxygenation( H/R) injury in human umbili-cal vein endothelial cells ( HUVEC ) and its mecha-nisms. Methods HUVECs cells were divided into five groups: normal ( control) group, S1P low concentra-tion group ( L ) , S1 P medium concentration group (M), S1P high concentration group ( H) and H/R group. MTT method was used to measure cell survival. Using flow cytometric analysis, the rate of cell apopto-sis was determined. The activities of total superoxide dismutase ( T-SOD) , copper/zinc superoxide dismuta-se ( CuZn-SOD ) , manganese superoxide dismutase ( Mn-SOD) activity, nitric oxide ( NO) and malondial-dehyde ( MDA ) content in cell culture medium were measured with colorimetry. Mitochondrial membrane potential in cells was observed with fluorescence micro-scope. Bax/Bcl-2, eNOS protein expression levels in HUVECs cells were observed with Western blot. Re-sults Compared with H/R group, S1P low, medium and high concentrations in the intervention group could significantly increase the cell survival rate after H/R injury, and increase activity of T-SOD, CuZn-SOD, Mn-SOD and decrease content of MDA. Moreover, S1 P could significantly increase NO content and in-crease eNOS protein expression, decrease apoptosis rate and inhibit the reduction of mitochondrial mem-brane potential. Conclusions S1P can decrease cell apoptosis rate of HUVECs after H/R injury with a cer-tain concentration dependence. The protection of S1P for cell apoptosis of HUVECs after H/R injury may be related to decreasing the intracellular MDA content and improving intracellular SOD activity, increasing mito-chondrial membrane potential and enhancing expres-sion of Bcl-2, anti-apoptotic protein.

Aim To study the protective effects of sphingosine 1-phosphate (S1P) postconditioning on rat myocardial cells injured by hypoxia/reoxygenation in reperfusion injury salvage kinase ( RISK ) signal path-way. Methods The cultured rat H9c2 cells were ran-domly divided into seven groups: ( 1 ) control group;(2) hypoxia/reoxygenation (H/R) group; (3) S1P group;(4) S1P+LY294002 group(S1P+LY); (5) LY group; ( 6 ) S1 P +PD98059 group ( S1 P +PD );(7) PD group. The viability of H9c2 cells was detec-ted using MTT method. The content of MDA in the cultured medium and the activity of T-SOD and Mn-SOD were measured with colorimetry. The concentra-tion of intracellular free calcium ion was detected by confocal microscopy. The rate of cell apoptosis was de-termined by flow cytometric analysis. Western blot was used to assess phosphorylation of Akt and ERK1/2 in H9c2 cells. Results Compared with the H/R group, S1P significantly increased vaibility of cells, lowered the rate of apoptosis, decreased the content of MDA in the culture medium, increased the activity of T-SOD and Mn-SOD, reduced concentration of intracellular calcium and increased the phosphorylation of Akt and ERK1/2 . When added LY294002 or PD98059 , the effects of S1P above were inhibited. Conclusion S1P protects H9 c2 cells against hypoxia/reoxygenation inju-ry. The protection of S1P was inhibited by LY294002, the inhibitor of PI3 K/Akt and PD98059 , the inhibitor of ERK1/2 . S1 P protects H9 c2 cells against hypoxia/reoxygenation injury via RISK pathway.

Objective To knock out Asxl2 gene in murine embryonic fibroblast cell line NIH3T3 using CRISPR/Cas9n system. Methods A pair of sgRNAs which targeted exon 5 of Asxl2 gene were designed and subcloned into the pX462 vec?tor. The recombined plasmids were verified by sequencing and transfected into NIH3T3 cell line. Single cells were isolated through serial dilutions, followed by an expansion period to obtain new monoclonal cell lines. The genomic DNA of the new monoclonal cell lines was extracted and a DNA fragment flanked the target site was amplified by genotyping PCR then se?quenced. Lastly, western blotting were applied to confirm whether Asxl2 was successfully knocked out. Results The CRIS?PR/Cas9n plasmids that targeted Asxl2 were successfully constructed. NIH3T3 cells were co-transfected with the two recom?binant constructs. After puromycin selection, subclonal cell lines were obtained and one of them was validated by genotyping PCR-sequencing. Western blotting also confirmed that Asxl2 was completely depleted in the NIH3T3 cell line. Conclu?sion CRISPR/Cas9n plasmids that targeted Asxl2 were successfully constructed therefore a Asxl2 knockout NIH3T3 stable cell line was established via this system.

OBJECTIVE:To study the inhibitory effect of dihydrotanshinone(DTS)on human lung cancer GLC-82 cell and its mechanism. METHODS:After treated with 0(blank control),5,10,20,40,80 and 100 μg/ml DTS for 24 and 48 h,MTT as-say was used to measure the inhibition rates and IC50 of cells;cell apoptosis was detected by flow cytometry after treated with 17.85 μg/ml DTS for 12,24 and 48 h to calculate apoptotic rate;Western blot was used to detect the protein expressions of Bcl-2, Bax and Caspase-3. RESULTS:Compared with blank control group,different concentrations of DTS inhibited the proliferation of cells;24 and 48 h maximal inhibition rate were 54.48% and 64.95%,respectively;IC50 were 62.36 and 33.94 μg/ml. DTS could induce cell apoptosis in positive time dependent manner,and the range of inhibition rate was 5.6%-29.6%;Western blot showed DTS could down-regulate the expression of Bcl-2 protein and up-regulate the expression of Caspase-3 protein (P<0.01 or P<0.05). CONCLUSIONS:DTS have significant inhibitory effect on GLC-82 cells and also induce cell apoptosis,by a possible mech-anism of down-regulating the expression of Bcl-2 protein and up-regulating the expression of Caspase-3 protein.

Objective To investigate the role of stromal cell-derived factor-1α(SDF-1α)/CXCR4 signal pathway in the therapeutic effects of hypoxic preconditioning endothelial progenitor cell (HEPC) transplantation on acute myocardial in-farction Methods Bone marrow endothelial progenitor cells (EPCs) were isolated from syngeneic adult male Wistar rats. EPCs were cultured under normoxic condition for 4 days and 1%O2+5%CO2+94%N2 condition for 3 days. The effect of HEP-Cs on the migration ability of 100μg/L SDF-1αwas observed. Western blot assay was used to detect the expression of CX-CR4, the solo receptor of SDF-1α on cells surface. Then, 26 syngeneic adult male Wistar rats were randomized into 3 groups:control group (n=8),EPCs group (n=9) and HEPCs group (n=9). The acute myocardium infarction animal model was established. At infarction, the rats received 5-points peri-infarct intramyocardial injections of PBS 200μL, 2×106 EPCs and 2 × 106 HEPCs. After 4 weeks, the haemodynamics parameters of cardiac function were analyzed by echocardiography. Results Compare with EPCs, the migration ability of HEPCs towards SDF-1α was increased significantly. The result of Western blot analysis showed an increased CXCR4 expression on the cell surface. After 4 weeks of transplantation, the left ventricular end systolic diameter and ejection fraction (EF%) were much improved in HEPCs group than those of EPCs group and control group (P＜0.05). Compare with control group, the left ventricular end-diastolic diameter was significantly im-proved in EPCs and HEPCs groups (P＜0.05). There was no significant difference in the improvement of the left ventricular end-diastolic diameter between HEPCs and EPCs groups (P>0.05). Conclusion SDF-1α/CXCR4 pathway was up-regu-lated by HEPCs, which showed the therapeutic effects via EPCs. The adjustment of SDF-1α/CXCR4 signaling pathway is an effective method for the treatment of ischemic heart diseases.

Objective To study the clinical efficacy of Off-Pump Coronary Artery Bypass Grafting (OPCABG) in the patients who have left main coronary artery stenosis and three-vessel lesion with low ejection fraction.MethodsFrom January 2010 to December 2010,28 cases with low ejection fraction,left main coronary artery stenosis and three-vessel lesion and received OPCABG were analyzed retrospectively.All cases underwent OPCABG under general anesthesia.The Hb,PLT,ALT,AST,BUN and Cr count before and 3 days after operation were recorded.LVEF andLVEDbeforeand 3monthsafter operationwererecorded.Thedata were compared.Results one case died of respiratory failure caused by pulmonary infection.The mortality rate was 3.75 ％.The value of six parameters are: Hb ( [ 137.94 ± 19.40 ],[ 111.06 ± 12.71 ] g/L),PLT ( [ 210.43 ±48.26 ] × 109/L,[ 148.4 ± 52.20 ] × 109/L),ALT ( [ 27.66 ± 11.51 ] U/L,[ 29.02 ± 16.40 ] U/L),AST ([26.55 ± 10.12] U/L,[27.75 ±8.14] U/L),BUN([6.51 ± 1.00] mmol/L,[6.88 ± 2.53] mmol/L),Cr ([96.35 ± 15.25]μmol/L,[95.11 ± 24.38 ] μmol/L).Comparison of the values showed that Hgb and PLT were different significantly before and 3 days after operations.There was no significant difference in the levels of ALT,AST,BUN and Cr before and after the treatment.Three months later,LVEF and LVED were significantly different from that before treatment ( P ＜ 0.05 ),with LVEF of( [ 36.23 ± 3.41 ] ％,[ 47.23 ± 5.13 ] ％ ) and LVED of( [62.23 ±5.93]mm,[58.08 ±5.02] mm).ConclusionOPCABG produces a satisfactory outcome in the patients who have left main coronary artery stenosis and three-vessel lesion combined with low ejection fraction.There is limited impact of OPCABG on the other organs.The long-term effect and prognosis are still need to be followed-up and analyzed.

ObjectiveTo access the predictive value in Postoperative mortality in patients undergoing coronary artery bypass grafting(CABG) by seven different Risk scoring system.MethodsTo collect the clinical information of patients undergoing CABG in our department.SinoSCORE,Additive EuroSCORE,Logistic EuroSCORE,OPR,Cleveland model,Parsonnet score,QMMI was used to predict postoperative mortality for all patients,and calculate the mean predictive postoperative mortality.To devided the patients to six group:group Ⅰ (0 - 1.99％ ),group Ⅱ (2.00％ - 3.99％ ),group Ⅲ (4.00％ - 5.99％ ),group Ⅳ ( 6.00％ - 7.99％ ),group Ⅴ (8.00％ - 9.99％ ),group Ⅵ ( ＞ 10％ ) by predictive postoperative mortality.Access the performance of risk scoring system predict the mortality through compare the predictive mortality and the observed mortality in different Risk stratification.To use Hosmer-Lemeshow goodness-of-fit test access the calibration.Discrimination was tested by determining the area under the receiver operating characteristic(ROC) curve(AUG).ResultsThe overall mean baseline age was ( 62.8 ± 8.8 ) years.The observed mortality in all our study patients was 1.99 ％ ( 22/1103 cases).The predictive mortality calculated by SinoSCORE,Additive EuroSCORE,Logistic EuroSCORE,OPR,Cleveland model,Parsonnet score and QMMI were 3.01％,4.38％,3.83％,1.69％,4.42％,6.71％ and3.71％.And the most closest is OPR,SinoSCORE,QMMI.Group tests confirmed Logistic EuroSCORE Overestimated the mortality in all the group.Predicted mortality calculated by Additive EuroSCORE was lower than the actual mortality in group Ⅵ and higher than the observed mortality in other group.SinoSCORE、Cleveland model、Parsonnet score、QMMI Overestimated the mortality in all the group expect group Ⅰ.OPR forecast a lower mortality Compared with observed mortality in group Ⅰ and group Ⅳ and a Slightly higher mortality in group Ⅱ,group Ⅲ.To use Hosmer - Lemeshow goodness-of-fit test access the calibration.The text proved all the risk scoring system had a good calibration for postoperative mortality (P ＞ 0.05 ).Discrimination was tested by ROC,only SinoSCORE ( AUC =0.751 ＞ 0.70) showed high discriminatory ability in predicting mortality.ConclusionSinoSCOBE have a good forecast performance for the postoperative mortality in the patients undergoing CABG in our department in seven different Risk scoring system.And SinoSCORE could be used in preoperative risk assessment.