Objective: To develop a RHCE genotyping assay based on kompetitive allele-specific PCR (KASP) and assess its clinical accuracy for RhCE blood group determination. Methods: KASP primers were designed to interrogate three RHCE loci: the 109 bp insertion/deletion in intron 2, c. 307T>C, and c. 676C>G. A total of 1 194 RhD-positive inpatients from Chengdu were typed by both KASP genotyping and manual tube serology. Discordant samples (n=10) were retested by both methods and further resolved by Sanger sequencing. An additional 377 cases were tested for the c. 48C>G locus to evaluate the predictive accuracy of individual loci and combined locus testing for RhC antigen. Results: Genotyping concordance with serology was 100.0% for both the c. 676C>G locus (RhE/Rhe) and the c. 307T>C locus (Rhc). For RhC prediction using the 109 bp insertion, overall accuracy was 99.7% (1 191/1 194); the 3 discordant cases were confirmed by Sanger sequencing to be false negatives attributable to 109 bp deletion in intron 2. Testing the c. 48C>G allele for RhC prediction yielded 7 false positives, with an accuracy of 98.1% (370/377). RhC antigen status was determined by combining the 109 bp insertion and the c. 48C allele. After excluding 10 samples with inconsistent results between the two loci, the accuracy reached 100% in the remaining 367 samples. When both loci were applied in combination, accuracy reached 100% in the 367 cases with concordant results. Among the 1 194 patients, CCee (45.8%) and CcEe (31.7%) were the most common RhCE phenotypes. The e antigen had the highest positivity rate (92.2%), and the Ce haplotype was the most frequent (66.9%). Conclusion: The KASP-based RHCE genotyping method achieves high accuracy for clinical RhCE typing. Combining the 109 bp insertion/deletion with the c. 48C allele significantly improves RhC antigen prediction compared with either locus alone. This method was applied to RhCE genotyping of 1 194 RhD-positive inpatients in Chengdu, providing local RhCE phenotype and haplotype distribution data to support RhCE-matched transfusion practice.

Immune thrombocytopenia (ITP) is a hemorrhagic autoimmune disease characterized by antibody-mediated platelet injury. ITP has complicated immunopathological mechanisms that need further elucidation. It is well known that the costimulatory molecules OX40 ligand (OX40L) and OX40 play essential roles in the immunological mechanisms of autoimmune diseases. Previously, we discovered that the expression of OX40L and OX40 is significantly increased in the peripheral blood mononuclear cells (PBMCs) of ITP patients. In our present study, OX40L-induced follicular helper T (Tfh) cells exhibited an activated phenotype with elevated expression of inducible T-cell costimulator (ICOS), programmed cell death protein-1 (PD-1), and cluster of differentiation 40 ligand (CD40L) in vitro. Moreover, aberrant OX40L‒OX40 expression might promote the Tfh1-to-Tfh2 shift in vivo, inducing the generation of autoantibodies by enhancing the helper function of Tfh cells for B lymphocytes in a mouse model, which might accelerate the progression of ITP. Additionally, signal transduction through the OX40L‒OX40 axis might be related to the activation of tumor necrosis factor receptor-associated factor (TRAF)‒nuclear factor-κB (NF-κB) and Janus kinase (JAK)‒signal transducer and activator of transcription (STAT) signaling pathways. Overall, OX40L‒OX40 signaling is proposed as a potential novel therapeutic target for ITP.
Animals ; OX40 Ligand/physiology* ; Purpura, Thrombocytopenic, Idiopathic/immunology* ; Cell Differentiation ; Mice ; T-Lymphocytes, Helper-Inducer/cytology* ; T Follicular Helper Cells/cytology* ; Signal Transduction ; Receptors, OX40 ; Mice, Inbred C57BL ; Humans ; Female

Objective:To study the revised content of the 2024 version of the Helsinki Declaration on vulnerability and the responsible inclusion of vulnerable participants, and to provide suggestions for improving regulations and ethics review in China. Methods:We analyzed the requirements of the Declaration of Helsinki and the preconditions for the inclusion of vulnerable groups, sorted out the international ethical principle and the relevant vulnerability provisions of China's laws and regulations, and proposed feasible recommendations for the inclusion and protection of vulnerable participants in China.Results:The Helsinki Declaration had been revised many times to involve the inclusion and protection of vulnerable participants, especially the Helsinki Declaration in 2024 for the first time proposed that vulnerability may originate from fixed or situational and dynamic factors, and vulnerability was no longer applicable to the traditional entire vulnerable group, and stipulated the prerequisites for the inclusion of vulnerable participants, because paying attention to " vulnerable groups" would affect the judgment of individual vulnerability, and should focus on the views and priorities of vulnerable individuals. Investigators and ethics committees should strengthen the identification and protection of vulnerable participants. The international ethical guidelines and China′s regulations had content on vulnerability, but there was a lack of consensus on the inclusion and protection of vulnerable participants. Conclusions:Feasible recommendations for the inclusion and protection of vulnerable participants in China include improving the relevant regulations on the inclusion of vulnerable participants in medical research, setting ethics review standards for vulnerable participants at the national level, strengthening the construction of ethics committees and training on the topic as well as clarifying key points of ethics review to maintain impartiality and avoid the misuse of vulnerable participants.

Objective:To examine the interaction between interpersonal issues and eating behavior problems among college freshmen, and to identify core psychological factors and potential pathways that drive eating behavior.Methods:In October 2019, a total of 5 073 college freshmen from a university in Shanghai were recruited as participants. Their general demographic data were collected, and they were evaluated using the eating disorder examination questionnaire 6.0 (EDE-Q 6.0) and inventory of interpersonal problems-32(IIP-32).Descriptive analyses were performed using SPSS 24.0 software.A regularized partial correlation network was constructed using the graphical least absolute shrinkage and selection operator (GLASSO) in R(v4.2.3) software. A Bayesian network analysis (BNA) was conducted to build a directed network, aiming to identify core driving factors and key psychological mechanisms.Results:Regularized partial correlation network identified shape concern as the most central node, with the highest strength centrality(1.32) and expected influence(1.20). It showed the highest bridge strength with dominance/control (0.22, 0.21), linking the interpersonal and eating behavior modules. Directed network analysis indicated that low self-confidence was the upstream node influencing social inhibition and cold/ distant relationships, indirectly affecting eating disorder.Within the eating module, shape concern and weight concern predicted eating preoccupation and dietary restraint, forming a pathway structure from eating cognition to behavior.The network demonstrated good stability (CS-coefficient=0.75).Conclusion:Shape concern is the core mechanism underlying eating problems in college freshmen. Low self-confidence contributes indirectly via interpersonal dysfunction. Dominance/control and shape concern bridge interpersonal and eating domains, providing key targets for early prevention and intervention.

Objective:Investigate the impact of calcification on the long-term outcomes of patients with coronary chronic total occlusion (CTO) after percutaneous coronary intervention (PCI).Methods:A retrospective cohort study was conducted. Patients who underwent PCI and had at least one CTO lesion at Fuwai Hospital between January 2010 and December 2013 were consecutively enrolled. Calcification was evaluated by coronary angiography, and patients were divided into two groups: moderate/severe calcification group and non/mild calcification group. Clinical follow-up was completed up to 5 years. Incidence of PCI-related complications and immediate procedural outcomes were compared between two groups, and the primary endpoint was the target lesion failure (TLF) at 5 years after PCI. Clinical follow-up endpoint events were analyzed using Kaplan-Meier survival analysis with log-rank test, and Cox multivariate regression model was used to evaluate the relationship between calcification and TLF.Results:The study included 2 659 CTO patients with an age of (57.2±10.5) years, of whom 442 (16.6%) were female, and among whom 13.5% (360/2 659) had moderate/severe calcification. Compared with the non/mild calcification group, the moderate/severe calcification group had a higher incidence of PCI-related complications (43.2% (156/361) vs. 32.5% (772/2 374), P<0.001) and procedural failure (34.3% (124/361) vs. 24.3% (577/2 374), P<0.001). Additionally, the moderate/severe calcification group showed a higher risk of the primary endpoint event (TLF) during the 5-year follow-up (19.8% vs. 15.3%, log-rank P=0.028). Higher incidence of cardiac death was observed in moderate/severe calcification group (5.7% vs. 2.7%, log-rank P=0.003). Cox multivariate regression analysis revealed that moderate/severe calcified plaques remained an independent risk factor for 5-year TLF after CTO-PCI ( HR=1.34, 95% CI: 1.01-1.79, P=0.043). Conclusion:Compared with CTO patients with non/mild calcification, those with moderate/severe calcification have higher procedural failure and complication rates, as well as poorer long-term prognosis, mainly due to an increase in cardiac death.

Temporal interference (TI) is a form of stimulation that epitomizes an innovative and non-invasive approach for profound neuromodulation of the brain, a technique that has been validated in mice. Yet, the thin cranial bone structure of mice has a marginal influence on the effect of the TI technique and may not effectively showcase its effectiveness in larger animals. Based on this, we carried out TI stimulation experiments on rats. Following the TI intervention, analysis of electrophysiological data and immunofluorescence staining indicated the generation of a stimulation focus within the nucleus accumbens (depth, 8.5 mm) in rats. Our findings affirm the viability of the TI methodology in the presence of thick cranial bones, furnishing efficacious parameters for profound stimulation with TI administered under such conditions. This experiment not only sheds light on the intervention effects of TI deep in the brain but also furnishes robust evidence in support of its prospective clinical utility.
Animals ; Deep Brain Stimulation/methods* ; Nucleus Accumbens/physiology* ; Male ; Rats ; Rats, Sprague-Dawley ; Time Factors

OBJECTIVES: This study aims to investigate factors influencing dental arch development in patients aged 0-6 years with cleft palate after Sommerlad-Furlow (SF) palatoplasty. METHODS: A total of 183 patients who underwent primary SF repair for cleft lip and palate before 18 months of age were included. Follow-ups were conducted at different ages, and digital dental casts of the maxillary dental arch were obtained using 3-matic Research 12.0 software. The length and width of the dental arch and palate were measured to explore developmental changes in the maxillary dental arch of the patients after the procedure. The study also investigated the influence of gender, age, cleft palate type, and relaxation incision on maxillary dental arch development. RESULTS: After SF, maxillary dental arch measurements showed statistically significant differences between children aged 0-2 years and those aged 3-6 years (P<0.05). However, no statistically significant differences were observed among different age groups within the 3-6 years range. Statistically significant differences were detected between males and females, with males having greater width of the posterior dental arch and palate (P=0.001) and shorter length of the anterior dental arch and entire dental arch (P<0.05). The unilateral cleft lip and palate group had shorter dental arch length (P<0.01) and wider posterior palate (P<0.01) than the cleft palate only group. Maxillary dental arch measurements had no statistically significant differences between groups with or without a relaxing incision. CONCLUSIONS Gender and age influence the width of the maxillary dental arch in children aged 0-6 years after SF, while age and cleft palate type affect dental arch length.
Humans ; Child, Preschool ; Male ; Cleft Palate/surgery* ; Female ; Child ; Infant ; Dental Arch/growth & development* ; Maxilla/growth & development* ; Cleft Lip/surgery* ; Age Factors ; Sex Factors ; Palate/surgery* ; Infant, Newborn

Based on the viewpoints of turbidity and toxin and years of clinical experience,professor Wei Aisheng proposed that spleen qi deficiency leading to internal retention of turbidity and toxin contributes to the key pathogenesis of patients with type 2 diabetes mellitus(T2DM)characterized by obesity or metabolic disorders.For T2DM patients with obesity or metabolic disorders in the middle and end stages,long-term retention of turbidity and toxin leads to unsmooth qi movement of triple energizer and damage of viscera.Therefore,Professor Wei put forward the viewpoint of cutting off the toxin in the early stage,and established the therapy of replenishing qi,respectively expelling pathogens and eliminating turbidity from triple energizer.Professor Wei formulated Sanhuang Yinchen Wuling San as the primary prescription for T2DM.The prescription is derived from the classical formula Yinchen Wuling San with the herbs of Astragali Radix,Coptidis Rhizoma and Ephedrae Herba added,having the actions of replenishing qi and supporting healthy qi,respectively expelling pathogens from triple-energizer to resolve phlegm-damp and stagnated heat.The application of Sanhuang Yinchen Wuling San for T2DM can invigorate spleen qi,smooth qi movement of triple energizer,and eliminate turbidity and toxin,and enhance the efficacy for T2DM.

Objective To investigate the efficacy of Lishui Xiaozhong Granules combined with Conbercept for the treatment of diabetic macular edema(DME).Methods From December 2022 to June 2023,46 patients(46 eyes)with DME of spleen and kidney yang deficiency complicated with blood-stasis obstructing the eye collaterals syndrome admitted to Foshan Hospital of Traditional Chinese Medicine were selected as the study objects.According to the principle of single-blind randomized controlled trial,the patients were randomly divided into treatment group and control group by random number table method,with 23 cases(23 eyes)in each group.The control group was treated with intravitreal injection of Conbercept,and the treatment group was treated with oral use of Lishui Xiaozhong Granules on the basis of treatment for the control group.The course of treatment lasted 3 months and then a follow-up for 6 months was conducted.Before and after treatment,the two groups were observed int the changes of traditional Chinese medicine(TCM)syndrome score,best corrected visual acuity(BCVA,LogMAR),central macular thickness(CMT),mean vessel density(VD)of superficial capillary plexus(SCP)and deep capillary plexus(DCP)in macular area,and the amplitude density and the latency of P1 and N1 wave in the R1 ring of multifocal electroretinogram(mf-ERG).Results(1)After 1,3 and 6 month(s)of treatment,BVCA in the two groups was improved when compared with that at the previous observation time point(P＜0.05),and the improvement in the treatment group was superior to that in the control group after 1,3 and 6 month(s)of treatment(P＜0.01).(2)After 1,3 and 6 month(s)of treatment,the CMT of the two groups was lowered as compared with that at the previous observation time point(P＜0.05),and the decrease in the treatment group was significantly superior to that in the control group(P＜0.01).(3)After 6 months of treatment,SCP-VD and DCP-VD in the two groups showed no obvious changes when compared with those before treatment(P＞0.05),the differences between the two groups after treatment were no statistically significant,either(P＞0.05).(4)After 1,3 and 6 month(s)of treatment,the amplitude densities of P1 wave and N1 wave in the R1 ring of mf-ERG in the two groups were increased when compared with those at the previous observation time point(P＜0.05),and the latency of P1 wave and N1 wave was shortened when compared with that at the previous observation time point(P＜0.05),and the treatment group had stronger effect on increasing the amplitude densities and on shortening the latency than the control group(P＜0.01).(5)After 6 months of treatment,the scores of TCM syndromes in the treatment group were lowered(P＜0.05)while the scores in the control group showed no obvious changes as compared with those before treatment.The intergroup comparison showed that the decrease of TCM syndrome score in the treatment group was significantly superior to that in the control group(P＜0.01).Conclusion In the treatment of DME with spleen and kidney yang deficiency complicated with blood-stasis obstructing the eye collaterals syndrome,Lishui Xiaozhong Granules combined with Conbercept have no influence on improving the SCP-VD and DCP-VD,but the combined therapy exerts satisfactory efficacy on increasing BCVA and on improving macular oedema,retinal function in the macular area and systemic symptoms.

Objective:To observe the clinical efficacy of gentle moxibustion at different temperatures in treating people with diarrhea-predominant bowel syndrome(IBS-D)due to spleen deficiency.Methods:A total of 108 IBS-D patients were divided into two groups using the random number table method,with 54 participants in each group.Moxibustion group 1 received gentle moxibustion at(43±1)℃at bilateral Tianshu(ST25)and Zusanli(ST36),lasting 30 min each session;moxibustion group 2 received gentle moxibustion at(37±1)℃at the same points.Both groups received the intervention 3 times weekly for a total of 18 sessions.Abdominal pain intensity,stool form,pattern-based efficacy,quality of life,and mental health assessments were performed at weeks 0,3,6,and 8.Results:The total effective rate for abdominal pain intensity was 87.8%in moxibustion group 1 versus 51.1%in moxibustion group 2,and the difference was statistically significant(P<0.05).When the treatment finished,abdominal pain intensity,the Bristol score,IBS-symptom severity scale(IBS-SSS)score,self-rating anxiety scale(SAS)score,and self-rating depression scale(SDS)score dropped significantly in both groups(P<0.05),and the IBS-quality of life(IBS-QOL)score increased markedly(P<0.05).Between-group comparisons demonstrated that abdominal pain intensity,the Bristol general score,IBS-SSS score,traditional Chinese medicine(TCM)pattern score,and SDS score were significantly lower in moxibustion group 1 than in moxibustion group 2 at treatment week 6(P<0.05),and the IBS-QOL score was notably higher in moxibustion group 1(P<0.05).Conclusion:Whether at 43℃or 37℃,gentle moxibustion at Tianshu(ST25)and Zusanli(ST36)can improve abdominal pain,stool form,and quality of life,reduce disease severity,and mitigate TCM pattern in IBS-D patients;43℃gentle moxibustion performs better than 37℃gentle moxibustion in improving abdominal pain,stool form,disease severity,TCM pattern,quality of life,anxiety,and depression in IBS-D.