BACKGROUND:LncRNA-TNFRSF13C,an important factor in B cell development and function,is expressed in periodontal tissues of patients with periodontitis,but the specific mechanism is still unclear. OBJECTIVE:To investigate the mechanism of lncRNA-TNFRSF13C regulating miR-1246 on hypoxia-inducible factor 1α in periodontal cells. METHODS:Human periodontal ligament cells(hPDLCs)were treated with lipopolysaccharide and divided into group A(hPDLCs cell lines without transfection),group B(hPDLCs cell lines transfected with TNFRSF13C NC-siRNA),group C(hPDLCs cell lines transfected with TNFRSF13C-siRNA),group D(hPDLCs cell line transfected with miR-1246 mimics),group E(hPDLCs cell line transfected with miR-1246 siRNA),group F(hPDLCs cell line transfected with TNFRSF13C-siRNA+miR-1246 mimics),and group G(hPDLCs cell line transfected with TNFRSF13C-siRNA+miR-1246 siRNA).The relative expression of lncRNA-TNFRSF13C and miR-1246 in each group was detected by qRT-PCR.Cell counting kit-8 assay was used to detect cell viability.Apoptosis was detected by flow cytometry.Expression of hypoxia-inducible factor 1α and vascular endothelial growth factor proteins was detected by western blot.The correlation between lncRNA-TNFRSF13C and miR-1246 was analyzed by Pearson,and the targeting relationship was analyzed by dual-luciferase reporter assay. RESULTS AND CONCLUSION:There was no significant difference in human periodontal ligament cell activity,apoptosis rate and protein indexes between groups A and B(P>0.05).Compared with group B,hPDLCS cell activity in group C was increased,and apoptosis rate and the expression of hypoxia-inducible factor 1α and vascular endothelial growth factor proteins were decreased(P<0.05).Compared with group C,hPDLCS cell activity in group D was decreased,and apoptosis rate and the expression of hypoxia-inducible factor 1α and vascular endothelial growth factor proteins were increased(P<0.05).Compared with group D,the cell activity of group E was increased(P<0.05).The cell activity in group F was lower than that in group E,and the apoptosis rate was reduced in both groups E and F(P<0.05).Compared with group F,the cell activity of group G was increased,and the apoptosis rate and the expression of hypoxia-inducible factor 1α and vascular endothelial growth factor were decreased(P<0.05).LncRNA-TNFRSF13C was positively correlated with miR-1246(P<0.05).Compared with the TNFRSF13C-siRNA group,the fluorescence activity of miR-1246-wt in the TNFRSF13C-NC group was reduced(P>0.05);compared with the miR-1246-NC group,the fluorescence activities of hypoxia-inducible factor 1α-wt and vascular endothelial growth factor-wt in the miR-1246 mimics group were increased(P<0.05).To conclude,down-regulation of lncRNA-TNFRSF13C can promote the activity of periodontal cells treated with lipopolysaccharide,reduce apoptosis,and inhibit hypoxia-inducible factor 1α and vascular endothelial growth factor.The mechanism is related to the regulation of miR-1246 activity.

Objective: To analyze the demographic characteristics of patients with red blood cell transfusion reactions, the usage of red blood cell preparations, and the differences in the composition ratio of adverse reactions based on multi-center data from the Haemovigilance Network, in order to reveal the clinical characteristics of red blood cell transfusion and its underlying issues. Methods: Clinical data of patients who experienced transfusion reactions after red blood cell transfusion in the Haemovigilance Network from 2018 to 2023 were collected. The demographic characteristics of patients who experienced transfusion reactions with different types of red blood cell preparations, the utilization of these preparations, and the differences of the composition ratios of transfusion reactions were analyzed. Count data were expressed as numbers (n) or percentages (%), and comparisons between groups were performed using the Chi-square test. Results: Red blood cell transfusion reactions were more common in females (53.56%), with the majority of patients aged 50-69 years (35.54%). The Han polulation accounted for the vast majority of patients (92.77%), and patients in the hematology and obstetrics/gynecology departments had a relatively high proportion of transfusion reactions (13.26% and 14.26%, respectively). Leukocyte-reduced red blood cells and suspended red blood cells were the most common types of transfusion reactions reported among red blood cell preparations. Allergic reactions and non-hemolytic febrile reactions were the most common transfusion reactions, and there were significant differences in the composition ratios of allergic reactions (χ =869.89, P<0.05) and non-hemolytic febrile reactions (χ =812.75, P<0.05) across various types of red blood cell preparations. Conclusion: There are differences in the demographic characteristics and composition ratio of transfusion reactions among different red blood cell preparations. The management of red blood cell transfusion reactions should be tailored to patient characteristics and conditions, and the selection and use of blood products should be optimized to reduce or avoid the occurrence of transfusion reactions, such as considering the use of washed red blood cells for patients with a history of transfusion allergies or those prone to allergies.

Objective : To investigate the biomechanical effect of alveolar bone graft (ABG) resorption on the maxillary alveolar process under occlusal force in a patient with unilateral cleft lip and palate (UCLP) and provide evidence for the clinical application of ABG. Methods: A 3D finite element maxillary model of an 11-year-old female patient with UCLP was generated. The occlusal force was applied to six models with different ABG resorption, namely non-resorption, upper 1/3 resorption, upper 2/3 resorption, lower 1/3 resorption, lower 2/3 resorption, and upper&lower 1/3 resorption. The properties of structures in all models were set to be linear, elastic, and isotropic. The displacement and Von Mises stress of each reference node of the alveolar process were compared and analyzed. Results: Under occlusal force, the most significant displacement of the alveolar process was located in the anterior area, and it decreased gradually from anterior area to both sides in all groups. The displacement values of the alveolar process under cleft side lateral occlusion were as follows: non-resorption group < lower 2/3 resorption group < upper&lower 1/3 resorption group < lower 1/3 resorption group < upper 2/3 resorption group < upper 1/3 resorption group. The displacement values of the alveolar process under centric occlusion were as follows: non-resorption group < lower 1/3 resorption group < upper&lower 1/3 resorption group < upper 2/3 resorption group < lower 2/3 resorption group < upper 1/3 resorption group. The displacement values of the alveolar process under non-cleft side lateral occlusion were as follows: non-resorption group < lower 1/3 resorption group < upper 1/3 resorption group < lower 2/3 resorption group < upper&lower 1/3 resorption group < upper 2/3 resorption group. The stress was concentrated on the premolar area on the functional side of the alveolar process, followed by the canine and molar areas in all groups. The stress values of the alveolar process under cleft side lateral occlusion were as follows: non-resorption group < lower 2/3 resorption group < upper&lower 1/3 resorption group < upper 2/3 resorption group < lower 1/3 resorption group < upper 1/3 resorption group. The stress values of the alveolar process under centric occlusion were as follows: non-resorption group < upper 1/3 resorption group < lower 1/3 resorption group < lower 2/3 resorption group < upper&lower 1/3 resorption group < upper 2/3 resorption group. The stress values of the alveolar process under non-cleft side lateral occlusion were as follows: non-resorption group < lower 2/3 resorption group < upper&lower 1/3 resorption group < lower 1/3 resorption group < upper 2/3 resorption group < upper 1/3 resorption group. Under occlusal force, the displacement and stress of the alveolar process in the non-resorption model were significantly lower than those in other models. The displacement and stress of the alveolar process in the models with resorption in the lower area of the ABG were significantly lower than those in the models with resorption in the upper-middle areas of the ABG. Conclusion After unilateral complete cleft lip and palate bone grafting, the integrity and continuity of the middle and upper parts of the alveolar process bone grafting play a key role in the biomechanical status of the alveolar process. If bone resorption occurs in the above parts, bone grafting should be considered.

OBJECTIVE To explore the effect of iron sulfur cluster binding protein 1(ISCA1)mag-netic field on calcium inflow.METHODS ① A plasmid containing the ISCA1、Magneto 2.0 gene sequence,and an empty plasmid was prepared for lentivirus packaging,with all the three plasmids carrying the mCherry gene.The infection efficiency of lentiviruses was assessed using fluorescence microscopy.② The immunoprecipitation technique was employed to ascertain the interactions between ISCA1 and cryptochrome(CRY1)as well as CRY2 proteins.③ The technique of live cell calcium imaging was used to detect the intracellular calcium influx in cells exhibiting high levels of ISCA1 or Magneto 2.0 expression under the influence of a magnetic field(40 mT,0.1Hz,90%duty cycle).RESULTS ① Red fluorescence was observed by fluorescence microscopy,indicating successful transfection of lentivirus.② The exogenous ISCA1 proteins exhibited no affinity towards the endogenous CRY1 or CRY2 proteins.③ The green fluorescence intensity of the Magneto 2.0 group increased by(1.8±0.5)times compared to the pre-magnetic addition level,indicating a significant influx of calcium ions.However,there was no significant difference in the green fluorescence intensity between the ISCA1 group and control group before and after magnetic addition.CONCLUSION Cells with exogenously high expres-sion of ISCA1 do not elicit calcium influx and exhibit no discernible magnetic induction following stimula-tion by this magnetic field.

Objective To construct models of viral transfection and hypoxia/reoxygenation induced cellular injury in adult mouse cardiomyocytes(AMCMs)isolated using a non-Langendorff method.Methods AMCMs were isolated,extracted,sedimented,and plated using a non-Langendorff method.The morphology and survival rate of the isolated cells were evaluated 2,24,48 and 72 h after plating,and their integrity was observed by immunofluorescence staining for α-actinin.The isolated AMCMs were infected with adenoviruses carrying an RFP-expressing vector and fluorescence images were obtained at 36 and 48 h post-infection and used to calculate transfection efficiency.The cells were cultured under hypoxic conditions for 45 min,reoxygenated for 24 h,and then stained with propidium iodide(PI)to verify establishment of the hypoxia/reoxygenation injury model.Results The survival rates of AMCMs at 2,24 and 48 h after plating were comparable,but survival was significantly reduced at 72 h.The integrity of the AMCMs was good and＞80%of the cells were transfected with adenovirus at 48 h.After hypoxia/reoxygenation treatment,42%of cells were stained by PI,suggesting successful establishment of the AMCM injury model.Conclusions In this study,we developed a non-Langendorff method for the fast and easy isolation of AMCMs with high cell viability.The isolated cells can be efficiently infected with adenovirus and respond to hypoxia/reoxygenation injury.These findings provide a systematic method for isolating AMCMs and for applying gene modification and hypoxia/reoxygenation injury in these cells.

Objective:To investigate urinary pH distribution and its influencing factors in gout patients, to provide insights for individualized treatment.Methods:This is a retrospective study. The gout patients in the Gout Outpatient Department of the Affiliated Hospital of Qingdao University from September 2019 to August 2021 were collected. Clinical data were collected and relevant indicators were measured. The patients were divided into different groups according to urinary pH. Clinical characteristics and factors related to urinary pH were compared among the groups. SPSS 23.0 software was used.Results:A total of 2 553 patients were enrolled. There were significant statistical differences in age, body mass index, triglyceride, alanine aminotransferase(ALT), blood urea nitrogen, serum creatinine, estimated glomerular filtration rate(eGFR), blood uric acid, urinary uric acid/creatinine ratio, fraction excretion of uric acid(FEUA) among groups with different urinary pH( F were 5.114, 4.772, 7.170, 4.721, 13.603, 2.812, 3.422, 22.834, 18.230, 26.332, all P<0.05). Urinary uric acid and FEUA in acute group were higher than those in remission group( Z were -2.295, -3.528, both P<0.05). After adjusting for gender, age, eGFR, logistics regression analysis showed that body mass index, triglyceride, total cholesterol, ALT, blood uric acid, and blood urea nitrogen were still risk factors. Multivariate logistic regression analysis showed that triglyceride, blood uric acid, and blood urea nitrogen were independent risk factors associated with acid urine. Linear correlation analysis showed that urinary pH was negatively correlated with body mass index, triglyceride, total cholesterol, blood uric acid, fasting glucose, blood urea nitrogen, ALT( r were -0.079, -0.106, -0.051, -0.186, -0.040, -0.122, -0.051, all P<0.05), but positively correlated with eGFR( r=0.058, P=0.003). Conclusion:The overall urine pH levels in patients with primary gout are below normal reference. Several metabolic components are related to it. Triglyceride, blood uric acid, and blood urea nitrogen are independent risk factors of acidic urine. In clinical practice, attention should be paid to timely alkalization of urine to prevent complications.

Objective:To investigate the clinical phenotype and genetic characteristics of cerebral small vessel disease (CSVD) caused by heterozygous mutations in high-temperature requirement A serine peptidase 1 ( HTRA1) gene. Methods:Nine patients with HTRA1 gene related autosomal dominant CSVD diagnosed in the Departments of Geriatrics and Neurology,Xiangya Hospital of Central South University between January 2020 and December 2023 were selected. Their clinical data were collected, and the probands received genetic test using whole exome sequencing. The mutations were then verified in the family using Sanger sequencing, and their clinical and genetic characteristics were summarized. Results:Among the 9 patients with HTRA1 gene related autosomal dominant CSVD, the onset age was (51.1±9.5) years. Cognitive impairment, stroke onset, and gait disturbance were the most common clinical manifestations. Magnetic resonance imaging examination usually revealed diffuse white matter lesions, multiple lacunar cerebral infarction, multiple cerebral microbleeds, and brain atrophy, which were consistent with the radiological characteristics of CSVD. Most patients showed a decrease in Aβ42 levels and Aβ42/Aβ40 ratio in cerebrospinal fluid, and 2/4 of patients had an increase in phosphory protein tau levels. Seven heterozygous mutations in the HTRA1 gene were found in 9 patients, including two new heterozygous missense mutations, c.1160T>C(p.M387T) and c.569G>T(p. A190L). Conclusions:The clinical manifestations of HTRA1 gene-related autosomal dominant CSVD patients are mainly cognitive impairment, stroke and gait disturbance, and the imaging features are consistent with CVSD changes. HTRA1 gene c.1160T>C(p.M387T) and c.569G>T(p.A190L) heterozygous missense mutations are newly reported mutations, expanding the genetic mutation spectrum of this disease.

Objective:To construct a novel dual-specific antibody targeting human CD123 (CD123 DuAb) and study its effects in acute myeloid leukemia (AML) .Methods:Based on the variable region of the CD123 monoclonal antibody independently developed at our institution, the CD123 DuAb expression plasmid was constructed by molecular cloning and transfected into ExpiCHO-S cells to prepare the antibody protein. Through a series of in vitro experiments, its activation and proliferation effect on T cells, as well as the effect of promoting T-cell killing of AML cells, were verified.Results:① A novel CD123 DuAb plasmid targeting CD123 was successfully constructed and expressed in the Expi-CHO eukaryotic system. ②The CD123 DuAb could bind both CD3 on T cells and CD123 on CD123 + tumor cells. ③When T cells were co-cultured with MV4-11 cells with addition of the CD123 DuAb at a concentration of 1 nmol/L, the positive expression rates of CD69 and CD25 on T cells were 68.0% and 44.3%, respectively, which were significantly higher than those of the control group ( P<0.05). ④Co-culture with CD123 DuAb at 1 nmol/L promoted T-cell proliferation, and the absolute T-cell count increased from 5×10 5/ml to 3.2×10 6/ml on day 9, and CFSE fluorescence intensity decreased significantly. ⑤ With the increase in CD123 DuAb concentration in the culture system, T-cell exhaustion and apoptosis increased. When the CD123 DuAb was added at a concentration of 1 nmol/L to the culture system, the proportion of CD8 + PD-1 + LAG-3 + T cells was 10.90%, and the proportion of propidium iodide (PI) - Annexin Ⅴ + T cells and PI + Annexin Ⅴ + T cells was 18.27% and 11.43%, respectively, which were significantly higher than those in the control group ( P<0.05). ⑥ The CD123 DuAb significantly activated T cells, and the activation intensity was positively correlated with its concentration. The expression rate of CD107a on T cells reached 16.05% with 1 nmol/L CD123 DuAb, which was significantly higher than that of the control group ( P<0.05). ⑦The CD123 DuAb promoted cytokine secretion by T cells at a concentration of 1 nmol/L, and the concentration of IFN-γ and TNF-α in the supernatant of the co-culture system reached 193.8 pg/ml and 169.8 pg/ml, respectively, which was significantly higher than that of the control group ( P<0.05). ⑧When CD123 DuAb was added at a concentration of 1 nmol/L to the co-culture system of T cells and CD123 + tumor cells, the killing intensity of T cells significantly increased, and the residual rates of CD123 + MV4-11 cells, CD123 + Molm13 cells, and CD123 + THP-1 cells were 7.4%, 6.7%, and 14.6% on day 3, respectively, which were significantly lower than those in the control group ( P<0.05) . Conclusion:In this study, a novel CD123 DuAb was constructed and expressed. In vitro experiments verified that the DuAb binds to CD123 + tumor cells and T cells simultaneously, promotes T-cell activation and proliferation, and facilitates their anti-leukemia effect, which provides a basis for further clinical research.

Objective To explore the protective effect of wogonin on retinal ganglion cells(RGC)in mice with glau-coma and its related mechanism.Methods Ten(10 eyes)C57BL/6J mice were included in the control group,and 40(40 eyes)DBA/2J mice were selected to prepare glaucoma models,which were divided into the model group,experiment 1,2 and 3 groups,with 10 mice per group.Mice in the control and model groups were given normal saline intragastrically,and mice in the experiment 1,2 and 3 groups were intragastrically given 50,100 and 150 mg·kg-1 wogonin,respectively.The right eyes were selected as experimental eyes.The intraocular pressure(IOP)of mice was compared among the five groups before and after 2,4 and 8 weeks of intragastric administration.The RGC number and retinal thickness of mice in each group were compared after hematoxylin-eosin staining.The levels of superoxide dismutase(SOD),glutathione peroxidase(GSH-Px)and nitric oxide synthase(NOS)in the retinal tissues of mice were detected by colorimetry.The levels of inter-leukin(IL)-6,IL-1 β and tumor necrosis factor-α(TNF-α)in the retinal tissues of mice were detected by enzyme-linked im-munosorbent assay.The expression levels of nucleotide oligomerization domain-like receptor family 3(NLRP3),Caspase-1 and apoptosis-associated speck-like protein(ASC)in the retinal tissues of mice were detected by Western blot.Results Compared with the control group,IOP in the model group and the experiment 1,2 and 3 groups increased(all P＜0.05)before intragastric administration.After 2,4 and 8 weeks of intragastric administration,IOP in the model group and the ex-periment 1,2 and 3 groups increased compared with the control group(all P＜0.05);IOP in the experiment 1,2 and 3 groups decreased compared with the model group(all P＜0.05);IOP in the experiment 2 and 3 groups decreased compared with the experiment 1 group(both P＜0.05);IOP in the experiment 3 group decreased compared with the experiment 2 group(P＜0.05).Compared with the control group,the number of RGC,retinal thickness,SOD and GSH-Px levels in the model group and the experiment 1,2 and 3 groups decreased,while the expression levels of NOS,IL-6,IL-1 β,TNF-α,and NLRP3,Caspase-1 and ASC proteins increased,and the differences were statistically significant(all P＜0.05).Compared with the model group,the number of RGC,retinal thickness,SOD and GSH-Px levels in the experiment 1,2 and 3 groups increased,while the expression levels of NOS,IL-6,IL-1β,TNF-α,and NLRP3,Caspase-1 and ASC proteins decreased,and the differences were statistically significant(all P＜0.05).Compared with the experiment 1 group,the number of RGC,retinal thickness,SOD and GSH-Px levels in the experiment 2 and 3 groups increased,while the expression levels of NOS,IL-6,IL-1 β,TNF-α,and NLRP3,Caspase-1 and ASC proteins decreased(all P＜0.05);compared with the experi-ment 2 group,the number of RGC,retinal thickness,SOD and GSH-Px levels in the experiment 3 group increased,while the expression levels of NOS,IL-6,IL-1β,TNF-α,and NLRP3,Caspase-1 and ASC proteins decreased(all P＜0.05).Conclusion Wogonin can improve the retinal thickness of mice with glaucoma and alleviate retinal oxidative damage and inflammatory reaction,which may be related to inhibiting the activation of NLRP3 inflammasomes.

Patients with severe trauma require an extremely timely treatment and transfusion plays an irreplaceable role in the emergency treatment of such patients. An increasing number of evidence-based medicinal evidences and clinical practices suggest that patients with severe traumatic bleeding benefit from early transfusion of low-titer group O whole blood or hemostatic resuscitation with red blood cells, plasma and platelet of a balanced ratio. However, the current domestic mode of blood supply cannot fully meet the requirements of timely and effective blood transfusion for emergency treatment of patients with severe trauma in clinical practice. In order to solve the key problems in blood supply and blood transfusion strategies for emergency treatment of severe trauma, Branch of Clinical Transfusion Medicine of Chinese Medical Association, Group for Trauma Emergency Care and Multiple Injuries of Trauma Branch of Chinese Medical Association, Young Scholar Group of Disaster Medicine Branch of Chinese Medical Association organized domestic experts of blood transfusion medicine and trauma treatment to jointly formulate Chinese expert consensus on blood support mode and blood transfusion strategies for emergency treatment of severe trauma patients ( version 2024). Based on the evidence-based medical evidence and Delphi method of expert consultation and voting, 10 recommendations were put forward from two aspects of blood support mode and transfusion strategies, aiming to provide a reference for transfusion resuscitation in the emergency treatment of severe trauma and further improve the success rate of treatment of patients with severe trauma.