[Objective] To use Platelet Additive Solution Ⅲ M to suspend concentrated platelets and evaluate their quality at different storage periods, in order to investigate the optimal ratio of Ⅲ M to plasma in the medium for storing concentrated platelets. [Methods] Disposable plastic blood bags with platelet storage bags were used to collect whole blood from healthy voluntary blood donors, and concentrated platelets were collected by plasma-rich method, with a volume of about 50 mL and ≥4.0×10¹⁰ platelets contained in each bag. According to the Platelet Addictive Solution ⅢM/plasma volume ratio in the medium of suspended platelets, the platelets were divided into 3 groups: control group (plasma only), experimental group 1(Platelet Addictive Solution ⅢM/plasma volume ratio of 6.5∶3.5) and experimental group 2 (low plasma group, Platelet Addictive Solution ⅢM/plasma volume ratio of 9∶1), each group of 50 samples. Three groups of platelets were stored at (22±2) ℃ at a flat-bed shaker, and 5 mL were sampled by sterile connection at day 1, 3, 5 and 7 respectively to detect platelet count, pH value, lactate dehydrogenase, CD62P positive rate and Annexin V positive rate. All the data were analyzed with SPSS24.0 software. One-way ANOVA was employed to compare the differences among three groups. In order to pairwise comparisons between means of multiple samples, Bonferroni method was applied. [Results] With the extension of storage time, the platelet count decreased and the Annexin V positive rate increased in the 3 groups, and the difference of the 3 groups was not statistically significant (P>0.05). The pH value decreased in the 3 groups, with values at day 1, 3, 5 and 7 of 7.44±0.13 vs 7.44±0.14 vs 7.41±0.11, 7.31±0.68 vs 7.43±0.23 vs 7.22±0.12, 7.30±0.15 vs 7.42±0.14 vs 7.17±0.12, 7.29±0.33 vs 7.26±0.18 vs 7.04 ± 0.12, respectively. The pH decline in the control group and experiment group 1 was minor, with no statistically significant difference, but experiment group 2 showed relatively larger decreases in day 5 and day 7, with statistically significant difference (P<0.05). LDH concentrate was elevated in 3 groups (mmol/L), with values at day 1,3,5 and 7 of 169.62±99.33 vs 105.80±150.71 vs 77.14±105.38, 225.10±112.86 vs 116.00±72.77 vs 94.42±88.74, 249.42±79.55 vs 119.00±53.51 vs 118.35±80.39, 253.34±86.95 vs 147.71±90.71 vs 124.68±128.68 respectively. Compared with the control group, the difference was statistically significant (P<0.05). Experimental group1 had the smallest increase; CD62P positive rate increased in 3 groups (%), with values at day 1, 3, 5 and 7 of 26.22±11.74 vs 23.48±12.48 vs 40.49±11.86, 41.29±8.36 vs 33.53±25.64 vs 50.42±22.36, 59.59±10.13 vs 36.39±23.10 vs 50.94±20.50, 72.92±15.44 vs 55.54±23.65 vs 61.89±18.82 respectively. Compared with the control group, the increase in experiment group1 was smaller, and the difference was statistically significant (P<0.05). [Conclusion] Platelet Addictive Solution ⅢM/plasma volume ratio of 6.5∶3.5 is superior to traditional plasma in maintaining platelet quality during the in vitro preservation period of platelets.

Tumor cell-intrinsic programmed death-ligand 1 (PD-L1) signals mediate tumor initiation, progression and metastasis, but their effects in ameloblastoma (AM) have not been reported. In this comprehensive study, we observed marked upregulation of PD-L1 in AM tissues and revealed the robust correlation between elevated PD-L1 expression and increased tumor growth and recurrence rates. Notably, we found that PD-L1 overexpression markedly increased self-renewal capacity and promoted tumorigenic processes and invasion in hTERT⁺-AM cells, whereas genetic ablation of PD-L1 exerted opposing inhibitory effects. By performing high-resolution single-cell profiling and thorough immunohistochemical analyses in AM patients, we delineated the intricate cellular landscape and elucidated the mechanisms underlying the aggressive phenotype and unfavorable prognosis of these tumors. Our findings revealed that hTERT⁺-AM cells with upregulated PD-L1 expression exhibit increased proliferative potential and stem-like attributes and undergo partial epithelial‒mesenchymal transition. This phenotypic shift is induced by the activation of the PI3K-AKT-mTOR signaling axis; thus, this study revealed a crucial regulatory mechanism that fuels tumor growth and recurrence. Importantly, targeted inhibition of the PD-L1-PI3K-AKT-mTOR signaling axis significantly suppressed the growth of AM patient-derived tumor organoids, highlighting the potential of PD-L1 blockade as a promising therapeutic approach for AM.
Ameloblastoma/metabolism* ; Humans ; B7-H1 Antigen/metabolism* ; Neoplasm Recurrence, Local/pathology* ; Signal Transduction ; Cell Proliferation ; Up-Regulation ; TOR Serine-Threonine Kinases/metabolism* ; Proto-Oncogene Proteins c-akt/metabolism* ; Telomerase/metabolism* ; Jaw Neoplasms/metabolism* ; Epithelial-Mesenchymal Transition ; Animals ; Cell Line, Tumor ; Female ; Male

Acceleration of tooth movement during orthodontic treatment is challenging,with osteoclast-mediated bone resorption on the compressive side being the rate-limiting step.Recent studies have demonstrated that mechanoreceptors on the surface of monocytes/macrophages,especially adhesion G protein-coupled receptors(aGPCRs),play important roles in force sensing.However,its role in the regulation of osteoclast differentiation remains unclear.Herein,through single-cell analysis,we revealed that CD97,a novel mechanosensitive aGPCR,was expressed in macrophages.Compression upregulated CD97 expression and inhibited osteoclast differentiation;while knockdown of CD97 partially rescued osteoclast differentiation.It suggests that CD97 may be an important mechanosensitive receptor during osteoclast differentiation.RNA sequencing analysis showed that the Rap1a/ERK signalling pathway mediates the effects of CD97 on osteoclast differentiation under compression.Consistently,we clarified that administration of the Rap1a inhibitor GGTI298 increased osteoclast activity,thereby accelerating tooth movement.In conclusion,our results indicate that CD97 suppresses osteoclast differentiation through the Rap1a/ERK signalling pathway under orthodontic compressive force.

ObjectiveTo present the exploration and application of a prospective follow-up research method for acute infectious disease surveillance based on natural community populations， using COVID-19 infection as an example， and to provide a reference for improving the infectious disease surveillance and early warning system. MethodsA multi-stage probability proportional sampling method was employed to sample residents from all communities of 16 administrative districts in Shanghai， with households as the units. A cohort for acute infectious diseases based on natural community populations was established. The baseline survey was conducted for all cohort subjects， and COVID-19 antigen test kits were distributed. From December 21， 2022 to September 30， 2023， prospective follow-up monitoring of COVID-19 antigen and nucleic acid was carried out on the study subjects on a weekly basis. The baseline characteristics and follow-up information of the cohort subjects were described. ResultsThe cohort for acute infectious diseases included a total of 12 881 subjects， comprising 6 098 males （47.3%） and 6 783 females （52.7%）. The baseline survey revealed that 35.2% （4 540/12 881） of the subjects had a history of COVID-19 infection. During the follow-up period from December 21， 2022 to September 30， 2023， the average incidence density in the cohort was 0.61/person-year， with a higher incidence density in females （0.63/person-year） compared to males （0.59/person-year）. Individuals aged 60 and above （0.64/person-year） and those with underlying health conditions （0.67/person-year） had a higher incidence density. Healthcare workers showed a notably higher incidence density （0.84/person-year） than that in other occupational groups. As of September 30， 2023， a total of 340 subjects in the cohort experienced secondary infections， with a median interval of 170 days between the first and second infections. ConclusionThis study applies cohort study method to acute infectious disease surveillance， providing crucial data support for estimating infection rates and forecasting alerts for acute infectious diseases in the community. This method can be promoted and applied as a new approach for acute infectious disease surveillance.

The gene encoding regulatory subunit 15 A of protein phosphatase 1 produces a protein that is a universally present protein phosphatase in eukaryotic cells.In this study,genomic DNAs were extracted from the blood of 89 Chinese Holstein cows and were used as templates for PCR amplification of the target fragment of the PPP1R15A gene.The product was tested and a polymor-phic site,E3-250T＞A was found.The polymorphism of this side and its correlation with milk pro-duction traits in Chinese Holstein cattle were statistically analyzed using SPSS 23.0 software.The findings revealed three genotypes at this site:AA,AT and TT.Cows possessing the AT and TT genotypes exhibited significant differences(P＜0.01)in milk fat and solid non-fat content com-pared to those with the AA genotype.While no significant differences were noted for other milk production traits,including milk yield,protein,lactose,somatic cell count,blood urea nitrogen,and corrected milk.The identification of functional SNPs in the PPP1R15A gene provides a theoretical basis for further research and identification of causal variations in the cow PPP1R15A gene.

【Objective】 To investigate whether the blood donors' coagulation status may lead to apheresis platelet aggregation in vitro. 【Methods】 Thirty blood donors with aggregation in apheresis platelets collected by AMICUS blood cell separator no less than 3 times previously and occurred when the last time of apheresis donation were observed in aggregated group (referred to as the experimental group); Thirty donors without aggregation in apheresis platelets collected by AMICUS blood cell separator no less than 3 times were observed in the control group simultaneously. The basic platelet parameters in the two groups, including Plt, MPV, PDW, Pet, P-LCR were detected by automatic blood cell analyzer (BC-3000Plus), and thromboelastogram indexes including reaction time(R), kinetics time(K), kinetics of clot development(α), maximum amplitude (MA) and coagulation index(CI) were tested by Thrombosis elastography (TEG) before collection. With SPSS24.0 software, t test was used to compare the differences between the two groups. 【Results】 The CI value in experimental group was significantly different from that of the control group (0.48± 1.00 vs -0.99 ±1.96, P< 0.05), and there was no significant difference in all above basic platelet parameters and other TEG parameters (P>0.05 ) . 【Conclusion】 The coagulation status of blood donors may be an independent risk factor for the in vitro aggregation of apheresis platelets.

【Objective】 To discuss the influence of apheresis platelets donation mode transformation, from walk-in to appointment, on apheresis platelets donation, donor retention and donation service quality. 【Methods】 The comparative research method is used to compare the number of apheresis platelets donors, blood donation units, rate of first-time blood donation, rate of repeated blood donation, conversion rate of fixed whole blood donors and satisfaction rate before and after the transformation of donation model. Questionnaires were randomly distributed to apheresis platelets blood donors before and after the transformation to study the evaluation of appointment mode. 【Results】 In comparison with walk-in mode, the number of blood donors after adopting the appointment mode was 30 193, with 41.93% (8 920/21 273) increase; number of blood donations was 119 143, with 93.66% (57 622/61 521) increase; platelet donation was 212 717 treatment units, with 103.12% (107 990/104 727) increase; rate of repeated blood donation was 53.56% (16 172/30 193), with 15.43% increase; the number of first-time donors was 15 949, with 57.93% (5 850/10 099) increase; the conversion rate of fixed whole-blood donors was 37.86% (6 039/15 949), with 8.84% increasement; the satisfaction of appointment mode reached 99.81%, with significantly improved satisfaction with blood donation environment and waiting time. 【Conclusion】 The appointment mode of apheresis platelet donation has a promoting role in the increase of apheresis platelets donation, the improvement of solid blood donors and the quality of apheresis platelets donation services.

Objective:To analyze the expression levels of miR-15 and miR-29a in serum of patients with diabetic retinopathy (DR) and their diagnostic values for DR.Methods:155 patients (155 eyes) with type 2 diabetes mellitus (DM) were treated in our hospital from June 2016 to August 2018, according to the occurrence of retinopathy and the degree of retinopathy, the patients were divided into five groups: 50 cases of non-retinopathy group (NDR group), 56 cases of simple retinopathy group (SDR group), 49 cases of proliferative retinopathy group (PDR group) and another 50 healthy persons in the same period were selected as the control group. Triglyceride (TG), total cholesterol (TC), and fasting blood glucose (FPG) were measured. Levels of serum miR-15 and miR-29a were detected by quantitative reverse transcription polymerase chain reaction (qRT-PCR). Pearson correlation method was used to analyze the relationships between levels of serum miR-15 and miR-29a with clinical indicators in DR patients. Receiver operating characteristic curve (ROC) was used to evaluate the diagnostic values of miR-15 and miR-29a in DR.Results:The relative expression of miR-15 in serum of NDR group, SDR group, and PDR group were lower than that of control group ( P<0.05), and the relative expression of miR-15 in NDR group, SDR group, and PDR group decreased gradually ( P<0.05); the relative expression of miR-29a in serum of NDR group, SDR group, and PDR group were higher than that of control group ( P<0.05), and the mRNA relative expression of miR-29a in NDR group, SDR group, and PDR group increased gradually ( P<0.05); the expression of serum miR-15 in DR patients was negatively correlated with urinary albumin/creatinine (UACR), glycosylated hemoglobin (HbA1c) and the course of DM ( r=-0.732, -0.492, -0.589, P<0.05); the expression level of miR-29a was positively correlated with UACR, HbA1c and the course of DM ( r=0.744, 0.508, 0.556, P<0.05); the areas under the ROC curve (AUC) of miR-15 and miR-29a for DR diagnosis were 0.796 (95% CI: 0.724-0.857) and 0.677 (95% CI: 0.597-0.749), with diagnostic thresholds 0.63 and 1.11, sensitivities 84.9% and 53.8%, specificities 65.3% and 79.6%, repectively; miR-29a was a risk factor for DR, while miR-15 was a protective factor for DR. Conclusions:The expression of mir-15 and miR-29a in the serum of DR patients is decreased, which is related to the degree of retinopathy and can be used as biomarkers for early diagnosis of DR.

Objective To observe the effects of interfering Twist expression on glycolysis in pancreatic cancer PANC1 cells and explore the potential mechanism.Methods Twist targeting siRNA (siTWIST) was designed and synthesized,and pancreatic cancer PANC1 cells were transfected with siTWIST using liposome,with nonspecific siRNA (siNC) transfected cells as negative control group and untransfected cells as blank control group.RT-PCR and Western blot were used to detect the expression levels of Twist mRNA and protein,and Akt and p-Akt protein expression in different groups;cell proliferation was detected by MTT;and glycolysis key enzyme (pyruvate kinase and hexokinase) activity and the content of lactic acid in the supernatant of culture fluid were detected.Results Twist mRNA level in control group,siRNA group and siTwist group expression was 1.00 ± 0.09,1.01 ± 0.08 and 0.36 ± 0.02;Twist protein level was 0.41 ± 0.05,0.42 ± 0.04 and 0.12 ± 0.06;the mRNA and protein expression in siTWIST group was obviously lower than those in control group and the difference was statistically significant (P ＜ 0.05).The cell survival rate was (100.02± 9.36)％,(100.01 ± 10.25)％ and (59.32± 7.26)％,which in siTWIST group was obviously lower than that in control group;pyruvate kinase activity was (0.73 ± 0.05)U/mg,(0.72 ± 0.08) U/mg and (0.43 ± 0.05) U/mg;the activities of hexokinase were (4.98 ± 0.48) U/mg,(4.96 ± 0.52) U/mg and (2.54 ± 0.21) U/mg;the content of lactic acid was (20.36 ± 2.61) mmol/L,(20.64 ± 3.05) mmol/L and (9.48 ± 1.09) mmol/L;the ratio of p-Akt/Akt was 0.65 ± 0.04,0.63 ± 0.07,and 0.25 ± 0.04,which in siTWIST group was obviously lower than that in control group.Conclusions Interference with Twist expression could inhibit the proliferation and glycolysis pathway in pancreatic cancer cells,and the mechanism may be related to the Akt signaling pathway.

Objective To study the homology characteristics of clinicaly isolated and colonized linezolid(LZD)-resistant Enterococcus faecalis (E.faecalis) strains from a patient.Methods Ten E.faecalis strains (2 were isolated from urine specimens and 8 were from stool specimens) isolated from a patient with pulmonary infection were performed antimicrobial susceptibility testing, homology of E.faecalis was determined by pulsed-field gel electrophoresis (PFGE).Results Before and after patients received LZD therapy, 2 E.faecalis strains isolated form urine specimens were both resistant to LZD (MICs: 8 mg/mL, 16 mg/mL, respectively), among 8 strains from stool specimens (6 were isolated before therapy, and 2 were isolated after therapy), LZD susceptible, intermediate, and resistant strains were 4, 2, and 2 respectively(MICs: 0.25-12 mg/mL).10 strains of E.faecalis were homologous by PFGE typing.Conclusion In this case, the detection of E.faecalis from urinary tract and intestinal tract is homologous, which suggested that LZD-resistant Enterococcus may be colonized in vivo for a long time, and may be shift to cause bacterial infection.