Objective: To characterize the serological profile, RHD genetic spectrum, and their frequencies among pregnant women preliminary screened as RhD-negative and weak positive in Qingdao and surrounding areas, and correlate these findings with unexpected antibody detection results, thereby providing testing recommendations and suggestions for such individuals. Methods: Blood samples of pregnant women who were initially identified as RhD negative and weak positive in hospitals in Qingdao and surrounding areas over the past five years were collected. Different cloned IgG anti-D antibodies were used for RhD negative confirmation experiments. RHD genotyping was performed by combining PCR-SSP and Sanger sequencing. Unexpected antibody screening and identification were carried out using test tube method and microcolumn gel card. The immunologic status of newborns delivered by anti-D pregnant women was also tracked. Results: A total of 602 blood samples were collected from pregnant women initially identified as RhD-negative and weak positive. Among them, 569 (94.5%) were confirmed as RhD-negative in the RhD confirmation test, and 33 (5.5%) were D variant phenotype. Except for 4 cases where no definite mutations were found, gene analysis revealed 474 (78.7%) D-negative cases with 5 genotypes (RHD 01N.01, RHD 01N.03, RHD 01N.16, RHD 01N.05, and 1 new allele), 90 (15.0%) Del cases with 2 genotypes (RHD 01EL.01, and RHD 01EL.18), 23 (3.8%) weak D cases with 2 genotypes (RHD 15 and 1 new allele), and 11 (1.8%) partial D cases with 2 genotypes (RHD 06.03.01 and RHD 05.04). Anti-D and complex antibodies containing anti-D were detected in 96 RhD-negative and partial D pregnant women (15.9%). After injection of anti-D immunoglobulin, One O-type RhD-negative pregnant woman delivered a newborn with hyperbilirubinemia. The newborn was typed to be B Del, and anti-D was detected in both serum and eluate. Conclusion: The serological profiles, RHD gene types and frequencies among RhD negative pregnant women in Qingdao and surrounding areas are basically consistent with domestic published data. Pregnancy can stimulate anti-D production in D-negative and partial D individuals. However, anti-D antibody has not been detected in Del type pregnant women. Since anti-D immunoglobulin can binds to Del type red blood cells, its administration is not recommended for Del type pregnant women.

[Objective] To conduct serological identification and molecular mechanism study on a ambiguous ABO blood group. [Methods] Standard serological techniques were used for the forward and reverse typing of ABO blood type. ABO gene coding and regulatory regions were analyzed by PCR after DNA extraction. Monoclonal sequencing was used to detect the haplotypes of the DNA sequence, and bioinformatics analysis was applied to predict the possible translation outcomes of the mutated DNA sequence. [Results] The sample’s red blood cells showed mixed field agglutination with anti-A, and the serum agglutinated with B cells, exhibiting serological characteristics of subtype A. Direct sequencing and monoclonal sequencing analysis of the ABO gene confirmed one allele as O02, the other had a c.27delC mutation compared with A102, which could cause the translation sequence to terminate prematurely at the 19th amino acids. Analysis and prediction suggested that the mutation might affect the function of the transferase through mechanisms such as shifting the initiation codon, altering the reading frame and affecting the splice sites. [Conclusion] This case is a rare A subtype caused by the c.27delC variation, and the impact on the glycosyltransferase may involve multiple mechanisms, which require further research and exploration.

Objective: To establish a "regular blood donor red blood cell gene database"(hereafter referred to as the "database") by applying molecular biology techniques for red blood cell antigens genotyping and utilizing information technology software, and to determine the significance and application value of this "database" in precise red blood cell transfusion. Methods: Fifteen antigens [C, c, E, e, M, N, S, s, Fy (a), Fy (b), Jk (a), Jk (b), Le (a), Le (b), P1] across six blood group systems (RHCE, MNS, FY, JK, Lewis and P1PK) were detected among 9 426 regular blood donors using the TaqMan-MGB method combined with an improved U-shaped microplate approach. With the assistance of information technology software, the "database" was integrated into the overall inventory management system of the blood supply chain. This enabled comprehensive management of regular blood donor and patient information, test results, specific antigen screening for regular blood donors, graded antigen matching between donors and patients, and rare blood type donor records. Results: The TaqMan-MGB method successfully detected paired antigens (C/c, E/e, M/N, S/s, Fy /Fy , Jk /Jk ) within a single reaction well using a standardized PCR amplification protocol. This method provided a reliable testing solution for clinical institutions and empowered blood collection and supply organizations with high-throughput screening capabilities. In the blood supply chain, genotyped red blood cells accounted for 13.2% (721/5 462 U) of the total inventory, with 95.34% (348/365) originating from donors who donated two units of blood. Moreover, the “database” fulfilled 94.06% (443/471 U) of compatible transfusion requirements from medical institutions and effectively managed rare blood type donors. Conclusion: The establishment of the "database" facilitated the transition of blood compatibility testing from traditional serological methods to molecular biology-based gold standard techniques, significantly advancing the implementation of precise transfusion strategies based on multi-antigen matching between donors and patients.

Objective: To perform serological identification and molecular analysis of four samples with antibodies against high-frequency antigens, and to track the condition of newborns after delivery. Methods: Blood group serological tests were conducted using tube method, and unexpected antibody screening and identification were performed using polybrene and human globulin card. Gene haplotype analysis of blood group system was performed using PacBio third-generation sequencing. The DNA mutations were confirmed through Sanger sequencing. Results: Four samples showed normal blood types in common blood type systems. However, they were positive in all unexpected antibody screening and identification, with negative direct antiglobulin tests results. Third-generation sequencing revealed 3 cases of c.376C>T homozygous mutation and 1 case of c.421C>A homozygous mutation in the ABCG2 gene. Three pregnant women gave birth to four children, all of whom developed hyperbilirubinemia, accompanied by decreased red blood cell count and normal or low hemoglobin concentration. Conclusion: Four samples were obtained from individuals with the rare Jr(a-) blood group. Immunization during pregnancy led to the production of anti-Jr antibody, which may contribute to hyperbilirubinemia in newborns.

Objective To analyze the deployment of policy instruments and the distribution of stakeholder engagement in provincial policies on nursing assistant training and management.Methods The relevant policy texts on nursing assistant training and management were systematically searched and collected from the official websites of provincial governments and their direct departments,CNKI,and the PKU Law Database.A two-dimensional framework of policy instruments-stakeholder was constructed,and the content analysis was used to classify,encode and quantify policy clauses.Results The study encompassed 20 provincial-level nursing assistant training and management policies,yielding a total of 359 codes.Within the policy instruments dimension,environmental,supply,and demand instruments constituted 66.30％,28.97％,and 4.74％,respectively.In terms of stakeholders,the management-side accounted for 56.55％,providers and trainers for 18.11％each,partners for 5.01％,and demand-side for 2.23％.Both management-side and trainers engaged with 3 policy instruments,providers with 2,and partners and demand-side with one each.Conclusion In the provincial nursing assistant training and management policies,there are differences in the deployment of policy instruments,and the distribution of stakeholders is uneven.Managers should pay attention to publicity guidance and platform construction,improve incentive mechanisms and training programs,and innovate cooperation models with stakeholders,as well as strengthen communication and exchange.

Objective:To retrospectively evaluate the clinical efficacy and safety of total 3D laparo-scopic ileal ureters replacement for bilateral ureters combined with bladder augmentation in patients with post-radiotherapy long-segment bilateral ureteral strictures and contracted bladder.Methods:Clinical data of two patients(aged 72 and 54 years)with radiation-induced long-segment bilateral ureteral stric-tures and reduced bladder capacity,treated at the Sixth Affiliated Hospital of Jinan University from Octo-ber 2023 to June 2024,were analyzed.Both presented with bilateral flank pain,recurrent chills/fever,urinary frequency,and urgency.Preoperative ureteral stricture lengths were measured as follows:left 10.4 cm and right 8.7 cm in the first case;left 10.6 cm and right 11.7 cm in the second case.Bladder capacity assessed by nephrostomy-assisted antegrade urography was 90 mL and 130 mL respectively.Both underwent single-position,one-stage totally 3D laparoscopic bilateral ileal ureteral replacement and blad-der augmentation based on membrane anatomy principles,with regular postoperative follow-up.Results:Procedures were completed by the same experienced urologist.Operative times were 420 min and 355 min,with intraoperative blood loss of 50 mL(no transfusion required).Postoperative bowel function re-sumed at the end of 4.5 and 3 days.No major perioperative complications occurred.Ureteral stents were removed at 2 months postoperatively,with imaging showing improved hydronephrosis,unobstructed ure-teral drainage,symmetrical bladder morphology,and smooth walls.Postoperative bladder capacities were 230 mL and 250 mL.Follow-up durations were 10 and 8 months.Both patients experienced significant relief of flank pain and lower urinary tract symptoms.No complications(enteric fistula,urinary fistula,or metabolic acidosis)were observed.At the final follow-up,one patient had mildly elevated serum cre-atinine,while the other showed reduced levels compared with preoperative values;both remained stable.Conclusion:Membrane anatomy-based dissection facilitates safe mobilization of fibrotic ureters with mini-mal bleeding and collateral damage.Total intracorporeal 3D laparoscopic ileal ureters replacement for bi-lateral ureters combined with bladder augmentation effectively addresses long-segment ureteral obstruction and improves bladder capacity.This approach is technically safe and feasible,though further validation with larger clinical cohorts is warranted.

Objective:To observe and analyze the correlation between the changes of macular microvascular structure and the level of intracocular fluid cytokines in patients with diabetic macular edema (DME).Methods:A prospective clinical study. From December 2022 to June 2024, 20 patients with 25 eyes of DME diagnosed by Department of Ophthalmology of Linyi People's Hospital were included in the study. Among them, 14 males had 17 eyes and 6 females had 8 eyes. Age was (55.08±10.34) years. Optical coherence tomography (OCT) and OCT angiography (OCTA) were used to scan the macular region at a range of 6 mm×6 mm. Central retinal thickness (CRT), blood flow density of superficial retinal capillary plexus (SCP) and area of fovea avascular zone (FAZ) were measured. The anterior aqueous humor was extracted before the first intravitreal injection of anti-vascular endothelial growth factor (VEGF), the concentrations of interleukin (IL-6), IL-8, VEGF, vascular cell adhesion molecule (VCAM), placental growth factor (PLGF) and monocyte chemotactic protein-1 (MCP-1) were detected. The correlation between macular microvascular structure and aqueous humor cytokines was analyzed by Spearman correlation analysis.Results:The CRT of the affected eyes was (617.40±167.64) μm, the SCP flow density was (39.56±1.55)%, and the FAZ area was (0.46±0.13) mm 2. The concentrations of IL-6, IL-8, VEGF, VCAM, PLGF and MCP-1 in aqueous humor were (301.36±690.52), (29.15±20.56), (71.37±29.32) and (5 621.22±7 241.06), (72.40±13.43), (464.07±163.26) pg/ml, respectively. Correlation analysis showed that there was a significant positive correlation between CRT and the concentrations of aqueous cytokines VEGF and PLGF ( r=0.460, 0.462, P<0.05). FAZ area was positively correlated with VEGF and MCP-1 concentrations ( r=0.414, 0.465; P<0.05). There was a significant negative correlation between SCP blood flow density and IL-6 ( r=0.401, P<0.05). Conclusion:There was a significant correlation between the morphological structure of macular area and the damage degree of microvessels around macular area in DME patients and the concentration of aqueous cytokines.

Objective:To investigate the prevalence of cardiometabolic multimorbidity (CMM) in Chinese middle-aged and elderly population, and the associated factors of cognitive function in CMM patients.Methods:It was a cross-sectional study. Data of demographic characteristics, lifestyle, chronic disease, and cognitive function in middle-aged and elderly subjects were obtained from the Harmonized CHARLS D version 2015 database, subjects with emotional and psychiatric disorders and memory-related disorders, and those with missing chronic disease data were excluded. The factors associated with cognitive function in CMM patients were analyzed with generalized linear regression model.Results:A total of 15 007 respondents aged (61.07±9.70) years were included in the analysis, 7 338 (48.9%) of whom were males. There were 3 303 cases of CMM with a prevalence rate of 22.01%, and the proportion of females was 54.9% (1 814/3 033), and the proportion of males was 45.1% (1 489/3 033). Generalized linear regression analysis showed that age, educational level, place of residence, depressive symptoms were correlated with cognitive function scores of CMM patients (all P<0.05). Conclusion:The prevalence rate of CMM in middle-aged and elderly people in China is higher, and age, education level, place of residence, depressive symptoms are associated with cognitive function in CMM patients.

Objective:To investigate the clinical efficacy of posterior midline incision combined with anteromedial approach in the treatment of complex olecranon fracture-dislocation.Methods:A retrospective analysis was performed on 26 patients (15 males and 11 females) with olecranon fracture-dislocation who were admitted from January 2020 to January 2024, including 5 cases of anterior transolecranon fracture-dislocation (2 cases of upper ulnar-radial joint dislocation), 21 cases of posterior transolecranon fracture-dislocation (5 cases of them were accompanied by upper ulnar-radial joint dislocation). Among them, there were 13 cases of traffic accidents, 7 cases of falling from heights, and 6 cases of walking falls. The average age is 45.1±15.3 years old (21-84 years old).Results:The operation time was 151.2±41.9 minutes, average tourniquet time was 93.7±22.6 minutes, and the intraoperative blood loss was 76.2±20.2 ml. The average follow-up was 16(12, 23) months, and the VAS score decreased significantly and the MEPS score increased significantly over time. At the last follow-up, the VAS score was 2(1, 2), and the MEPS score was 86.5±10.3, with 16 cases excellent, 7 cases good, and 3 cases medium, with an excellent rate of 89%. The range of motion of flexion-extension and pronation-supination were 119.3°±13.5°and 138.6°±15.2° respectively. Complications included 16 cases of ectopic ossification, of which 4 patients with significant effects on elbow function underwent surgical release 3-6 months after surgery. 1 case of ulnar nerve injury symptoms improved after emergency ulnar nerve release, and 1 case of elbow subluxation due to inaccurate coronoid process reduction and fixation. There were no serious complications such as vascular injury, internal fixation failure, fracture nonunion, and incision infection.Conclusion:The posterior midline incision combined with anteromedial approach can effectively treat complex olecranon fracture-dislocation and meet the requirements of early postoperative elbow rehabilitation.

Objective:To retrospectively evaluate the clinical efficacy and safety of total 3D laparo-scopic ileal ureters replacement for bilateral ureters combined with bladder augmentation in patients with post-radiotherapy long-segment bilateral ureteral strictures and contracted bladder.Methods:Clinical data of two patients(aged 72 and 54 years)with radiation-induced long-segment bilateral ureteral stric-tures and reduced bladder capacity,treated at the Sixth Affiliated Hospital of Jinan University from Octo-ber 2023 to June 2024,were analyzed.Both presented with bilateral flank pain,recurrent chills/fever,urinary frequency,and urgency.Preoperative ureteral stricture lengths were measured as follows:left 10.4 cm and right 8.7 cm in the first case;left 10.6 cm and right 11.7 cm in the second case.Bladder capacity assessed by nephrostomy-assisted antegrade urography was 90 mL and 130 mL respectively.Both underwent single-position,one-stage totally 3D laparoscopic bilateral ileal ureteral replacement and blad-der augmentation based on membrane anatomy principles,with regular postoperative follow-up.Results:Procedures were completed by the same experienced urologist.Operative times were 420 min and 355 min,with intraoperative blood loss of 50 mL(no transfusion required).Postoperative bowel function re-sumed at the end of 4.5 and 3 days.No major perioperative complications occurred.Ureteral stents were removed at 2 months postoperatively,with imaging showing improved hydronephrosis,unobstructed ure-teral drainage,symmetrical bladder morphology,and smooth walls.Postoperative bladder capacities were 230 mL and 250 mL.Follow-up durations were 10 and 8 months.Both patients experienced significant relief of flank pain and lower urinary tract symptoms.No complications(enteric fistula,urinary fistula,or metabolic acidosis)were observed.At the final follow-up,one patient had mildly elevated serum cre-atinine,while the other showed reduced levels compared with preoperative values;both remained stable.Conclusion:Membrane anatomy-based dissection facilitates safe mobilization of fibrotic ureters with mini-mal bleeding and collateral damage.Total intracorporeal 3D laparoscopic ileal ureters replacement for bi-lateral ureters combined with bladder augmentation effectively addresses long-segment ureteral obstruction and improves bladder capacity.This approach is technically safe and feasible,though further validation with larger clinical cohorts is warranted.