Epilepsy is a chronic neurological disorder affecting ~65 million individuals worldwide. Abnormal synaptic plasticity is one of the most important pathological features of this condition. We investigated how ubiquitin-specific peptidase 47 (USP47) influences synaptic plasticity and its link to epilepsy. We found that USP47 enhanced excitatory postsynaptic transmission and increased the density of total dendritic spines and the proportion of mature dendritic spines. Furthermore, USP47 inhibited the degradation of the ubiquitinated α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptor (AMPAR) subunit glutamate receptor 1 (GluR1), which is associated with synaptic plasticity. In addition, elevated levels of USP47 were found in epileptic mice, and USP47 knockdown reduced the frequency and duration of seizure-like events and alleviated epileptic seizures. To summarize, we present a new mechanism whereby USP47 regulates excitatory postsynaptic plasticity through the inhibition of ubiquitinated GluR1 degradation. Modulating USP47 may offer a potential approach for controlling seizures and modifying disease progression in future therapeutic strategies.
Animals ; Receptors, AMPA/metabolism* ; Neuronal Plasticity/physiology* ; Seizures/physiopathology* ; Disease Models, Animal ; Mice, Inbred C57BL ; Mice ; Ubiquitin Thiolesterase/genetics* ; Male ; Excitatory Postsynaptic Potentials/physiology* ; Ubiquitination ; Dendritic Spines/metabolism* ; Hippocampus/metabolism*

Objective:To explore the molecular mechanism of a case with RhD-phenotype.Methods:A proband with RhD-phenotype who attended the clinic of the First Affiliated Hospital of Zhengzhou University on January 29, 2024 was selected as the study subject. Peripheral blood samples were collected from the proband (8 mL) and her close relatives (father, mother and brother; 3 mL each) for Rh phenotyping and irregular antibodies testing with gel card and test tube methods. Direct agglutination reaction and absorption-elution test were used to detect the c antigen on the red blood cells of the proband. PCR-sequence specific primers (PCR-SSP) typing and gene sequencing were used to determine the RHCE gene of the proband and her relatives. The origin of the proband′s variant was traced by pedigree analysis. Three-dimensional structural models of the wild-type RhCE*cE protein and the RhD-phenotype protein were constructed to predict the alterations of the RhD-phenotype protein caused by the variant. The procedures of this study were approved by the Medical Ethics Committee of the First Affiliated Hospital of Zhengzhou University (Ethics No.: 2023-KY-0870-003). Results:The red blood cells of the proband did not agglutinate with anti-C, anti-c, anti-E, and anti-e. The result of the serum irregular antibody test was negative. The results of direct agglutination reaction and absorption-elution test of the proband were both negative. Her Rh blood group was identified as RhD-. The results of the Rh blood grouping of her close relatives were normal. PCR-SSP detection showed that the RHCE genotypes of the proband and her close relatives were cE/cE and Ce/cE, respectively. Gene sequencing analysis showed that the RHCE genotypes of the proband and her close relatives were RHCE* cE (c.365C>A)/ RHCE* cE (c.365C>A) and RHCE* Ce/ RHCE* cE (c.365C>A), respectively. Pedigree analysis revealed that the variants in the proband were inherited from her father and mother, respectively. Homology modeling of RhCE*cE protein showed that the RhD-type peptide chain with a significantly shortened C-terminal was encoded by only 121 amino acid resides, which was 296 amino acid resides shorter compared to the wild-type RhCE*cE peptide chain encoded by 417 amino acid residues. Conclusion:Above results revealed the molecular biological mechanism of a RhD-phenotype. The c. 365C>A variant in the RHCE gene has rendered the RHCE* cE alleles invalid, which ultimately led to the RhD-phenotype.

Objective:To investigate the impact of plasma exchange (PE) on T lymphocytes and natural killer (NK) cells in patients after kidney transplantation.Methods:The study retrospectively collected clinical data from patients who underwent their first ABO-compatible kidney transplantation at the Kidney Transplantation Center of the First Affiliated Hospital of Zhengzhou University between January 1, 2021, and November 30, 2023. Based on whether the patients received PE after transplantation, they were divided into a PE group and a non-PE group. A total of 38 patients were included in the retrospective cohort study, 18 in the PE group (8 males and 10 females) and 20 in the non-PE group (11 males and 9 females), with median ages of 37.5 (19.0, 56.0) years and 40.7 (22.0, 60.0) years, respectively. The immune cell subpopulations of the two groups were analyzed and compared at three time points: before PE, after PE, and 7 to10 days post-PE (for the non-PE group, data were collected at equivalent time points). The primary focus was on the proportions and absolute counts of T cells, NK cells, and activated T cells (CD4CD25+T cells, CD8CD25+T cells, and CD8HLA-DR+T cells). A generalized estimating equation (GEE) was used for longitudinal analysis of these repeated measurements to evaluate the effects of PE on changes in these cell populations.Results:From 7 to 10 days after PE, the absolute count of CD4 T cells in the non-PE group increased from 42 (24, 152) cells/μl to 461 (309, 608) cells/μl, while in the PE group, it increased from 57 (11, 262) cells/μl to 212 (141, 576) cells/μl. According to the generalized estimating model, the difference in changes between the two groups was statistically significant ( Z=-2.9, P=0.004). For NK cell absolute counts, the non-PE group increased from 20 (16, 36) cells/μl to 43 (26, 81) cells/μl, while the PE group increased from 22 (12, 63) cells/μl to 90 (28, 142) cells/μl. The difference in changes between the two groups was also statistically significant ( Z=1.87, P=0.049). Regarding T cell activation, the proportion of CD8HLA-DR+T cells in the non-PE group decreased from 25.8% (18.4%, 45.0%) to 22.7% (15.2%, 31.6%), while in the PE group, it increased from 19.2% (8.1%, 33.2%) to 22.7% (15.2%, 31.6%). The difference in changes between the two groups was statistically significant ( Z=2.88, P=0.005). From 7 to 10 days after PE, there were significant differences in the changes of CD4CD25+T cells ( Z=2.70, P=0.009), CD8CD25+T cells ( Z=2.75, P=0.007), and CD8HLA-DR+T cells ( Z=4.04, P=0.001) between the PE group and the non-PE group. Conclusion:Plasma exchange after kidney transplantation can suppress the proliferation of CD4 T lymphocytes, increase the number of NK cells relatively, and maintain a high proportion of activated T lymphocytes.

Objective:To investigate the effect of plasma exchange (PE) in improving renal function among patients with delayed graft function (DGF)following kidney transplantation.Methods:This was a retrospective cohort study. Data were collected from 76 patients who underwent their first-time ABO-compatible kidney transplantation at the Kidney Transplantation Center of the First Affiliated Hospital of Zhengzhou University between January 1, 2022, and November 30, 2023, and subsequently developed DGF and received PE treatment. The cohort included 39 males and 37 females, with a median age of 37.5 years (30.8-46.0 years). Patients were categorized into three groups based on pre-PEhuman leukocyte antigen (HLA) antibody status: HLA antibody-negative group, HLA antibody-unknown group, and HLA antibody-positive group. Additionally, immunological status prior to PE categorized patients into four groups: (1) stable lymphocyte subsetand antibody profiles (21 cases); (2) elevated B cell proportions or antibody production, or increased antibody titers (26 cases); (3) increased proportions of T/NK/macrophages (13 cases); (4) increased proportions of T/NK/macrophages combined with antibody production or increased antibody titers (16 cases). The Wilcoxon rank-sum test was used to analyze changes in serum creatinine (Scr) levels and 24-hour urine output before and after PE in the different patient groups.Results:In the HLA antibody-negative group, Scr decreased significantly from 467 (260, 571) μmol/L before PE to 176 (123, 307) μmol/L after PE ( Z=-2.22, P<0.01), and 24-hour urine output increased significantly from 1 295 (480, 2 020) ml to 1 960 (1 632, 2 870) ml ( Z=1.76, P<0.01). In the HLA antibody-positive group, Scr decreased significantly from 420 (254, 660) μmol/L before PE to 177 (151, 287) μmol/L after PE ( Z=-3.26, P<0.01), and 24-hour urine output increased significantly from 1 355 (928, 1 925) ml to 2 440 (1 760, 2 797) ml ( Z=2.47, P<0.01). For patients grouped by immunological status, Scr levels in Group 2 decreased significantly from 407 (242, 699) μmol/L to 201 (157, 274) μmol/L ( Z=-2.92, P<0.001), while in Group 3, Scr decreased significantly from 330 (258, 594) μmol/L to 164 (152, 280) μmol/L ( Z=-1.97, P=0.017). Regarding 24-hour urine output, Group 2 showed a significant increase from 1 353 (850, 1 770) ml to 1 995 (1 740, 2 630) ml ( Z=3.43, P=0.003), while in Group 3, urine output increased from 1 850 (1 350, 2 480) ml to 2 200 (1 900, 2 850) ml, but the difference was not statistically significant ( Z=1.10, P>0.05). Conclusion:PE effectively reduces Scr levels and increase 24-hour urine output in patients with DGF after kidney transplantation, regardless of pre-treatment HLA antibody status. Additionally, for patients with post-transplant changes primarily in T/NK/macrophages without significant antibody changes, PE can also effectively reduce Scr levels.

Objective: To reveal the molecular basis of the cisAB (p. Met266Leu) glycosyltransferase by studying a proband with cisAB subtype and his family. Methods: A male newborn was selected as the research subject. Tube methods were used to identify ABO blood types of the proband and his family members. PCR-SSP detection, ABO gene sequencing, and cloning analysis were performed on the proband and some family members. The inheritance pattern of the subtype gene in the family was determined through pedigree analysis. Homology modeling was used to analyze the impact of amino acid variations on the structure of the transferase, and molecular docking was used to demonstrate the bifunctional activity of the transferase and the donor-receptor binding conformation. Results: Serological tests showed that the proband and his father had enhanced anti-H agglutination, and the grandmother had a forward and reverse discrepancy. Sequencing of the proband revealed heterozygous variations of c. 297A>G, c. 526C>G, c. 657C>T, c. 703G>A, c. 803G>C, and c. 930G>A compared with A1. 01 (compared with B. 01, lacking the c. 796C>A variation, namely harboring the c. 796A>C variation) and c. 261delG. Combined with cloning analysis, the proband's genotype was determined to be ABO cisAB (c. 796A>C)/ABO O. 01. 01, the father's genotype was ABO cisAB (c. 796A>C)/ABO O. 01. 02, and the grandmother's genotype was ABO cisAB (c. 796A>C)/ABO B102. Pedigree analysis indicated that the cisAB allele in this newborn was inherited from his father and grandmother rather than a natural mutation. Homology modeling showed that the side chain orientation and intermolecular forces of Leu266 in the cisAB (p. Met266Leu) transferase changed, and molecular docking demonstrated that the "binding pocket" of the active center of the variant enzyme could accommodate both UDP-GalNAc and UDP-Gal, indicating that the cisAB enzyme structure has bifunctional activity. Conclusion: The bifunctional activity of this cisAB (p. Met266Leu) enzyme is related to the nucleotide variation of c. 796A>C, and molecular docking indicates that the enzyme has dual affinity for A/B sugar donors.

Abstract Breastfeeding has an irreplaceable effect on the growth and development of premature infants and the reduction of complications during hospitalization. In recent years, with the opening of China′s birth policy, the incidence of premature babies has increased, and the corresponding complications have also gradually increased. Breast milk not only carries a rich supply of nutrients, including rich proteins, lipids, sugars to support the normal growth and development of infants, but also contains a large number of immunoglobulins, hormones and other bioactive substances to promote the improvement of body immune function and the development of nervous system. In recent years, many studies have been carried out on the current situation of breast milk operation in breast milk bank, the collection and influencing factors of breast milk, the strategies of breast milk enrichment and its effects on the growth and development of premature infants and complications. Therefore, breast-feeding and intensive breast-feeding can effectively promote the growth and development of premature infants, and reduce the occurrence of complications.

OBJECTIVE: To explore the molecular mechanism of a case with RhD-- phenotype. METHODS: A proband with RhD-- phenotype who attended the clinic of the First Affiliated Hospital of Zhengzhou University on January 29, 2024 was selected as the study subject. Peripheral blood samples were collected from the proband (8 mL) and her close relatives (father, mother and brother; 3 mL each) for Rh phenotyping and irregular antibodies testing with gel card and test tube methods. Direct agglutination reaction and absorption-elution test were used to detect the c antigen on the red blood cells of the proband. PCR-sequence specific primers (PCR-SSP) typing and gene sequencing were used to determine the RHCE gene of the proband and her relatives. The origin of the proband's variant was traced by pedigree analysis. Three-dimensional structural models of the wild-type RhCE*cE protein and the RhD-- phenotype protein were constructed to predict the alterations of the RhD-- phenotype protein caused by the variant. The procedures of this study were approved by the Medical Ethics Committee of the First Affiliated Hospital of Zhengzhou University (Ethics No.: 2023-KY-0870-003). RESULTS: The red blood cells of the proband did not agglutinate with anti-C, anti-c, anti-E, and anti-e. The result of the serum irregular antibody test was negative. The results of direct agglutination reaction and absorption-elution test of the proband were both negative. Her Rh blood group was identified as RhD--. The results of the Rh blood grouping of her close relatives were normal. PCR-SSP detection showed that the RHCE genotypes of the proband and her close relatives were cE/cE and Ce/cE, respectively. Gene sequencing analysis showed that the RHCE genotypes of the proband and her close relatives were RHCE*cE (c.365C>A)/RHCE*cE (c.365C>A) and RHCE*Ce/RHCE*cE (c.365C>A), respectively. Pedigree analysis revealed that the variants in the proband were inherited from her father and mother, respectively. Homology modeling of RhCE*cE protein showed that the RhD-- type peptide chain with a significantly shortened C-terminal was encoded by only 121 amino acid resides, which was 296 amino acid resides shorter compared to the wild-type RhCE*cE peptide chain encoded by 417 amino acid residues. CONCLUSION Above results revealed the molecular biological mechanism of a RhD-- phenotype. The c.365C>A variant in the RHCE gene has rendered the RHCE*cE alleles invalid, which ultimately led to the RhD-- phenotype.
Humans ; Rh-Hr Blood-Group System/chemistry* ; Female ; Phenotype ; Male ; Alleles ; Pedigree ; Base Sequence ; Molecular Sequence Data ; Adult

[Objective] To study the molecular biological mechanism for a case of ABO blood group B subtype, and perform three-dimensional modeling of the mutant enzyme. [Methods] The ABO phenotype was identified by the tube method and microcolumn gel method; the ABO gene of the proband was detected by sequence-specific primer polymerase chain reaction (PCR-SSP), and the exon 6 and 7 of the ABO gene were sequenced and analyzed. Homologous modeling of Bw.03 glycosyltransferase (GT) was carried out by Modeller and analyzed by PyMOL2.5.0 software. [Results] The weakening B antigen was detected in the proband sample by forward typing, and anti-B antibody was detected by reverse typing. PCR-SSP detection showed B, O gene, and the sequencing results showed c.721 C>T mutation in exon 7 of the B gene, resulting in p. Arg 241 Trp. Compared with the wild type, the structure of Bw.03GT was partially changed, and the intermolecular force analysis showed that the original three hydrogen bonds at 241 position disappeared. [Conclusion] Blood group molecular biology examination is helpful for the accurate identification of ambiguous blood group. Homologous modeling more intuitively shows the key site for the weakening of Bw.03 GT activity. The intermolecular force analysis can explain the root cause of enzyme activity weakening.

Objective:To explore the trajectory and predictors of depressive symptoms in disabled middle-aged and elderly adults in China.Methods:Survey data from the 2013-2020 China Health and Retirement Longitudinal Study (CHARLS) data of disabled middle-aged and elderly adults were selected to analyze the trajectory categories of depressive symptom in disabled middle-aged and elderly adults and to explore their predictors.Results:A total of 1 844 disabled middle-aged and elderly adults were analyzed for survey data. The trajectory of depressive symptoms in disabled middle-aged and elderly adults was categorized into four groups, including a low-level depression group, a depression remission group, a high-level depression group, and a depression worsening group. Logistic regression analyses revealed that gender, age, place of residence, chronic disease, self-rated health, social participation, cognition, and hearing ability were predictors of trajectories of depressive symptoms in disabled middle-aged and elderly adults.Conclusions:There is group heterogeneity in the trajectory of depressive symptoms among disabled middle-aged and elderly adults. Healthcare professionals can carry out targeted health management according to the characteristics of different categories of people.

Objective:To explore the trajectory and predictors of depressive symptoms in disabled middle-aged and elderly adults in China.Methods:Survey data from the 2013-2020 China Health and Retirement Longitudinal Study (CHARLS) data of disabled middle-aged and elderly adults were selected to analyze the trajectory categories of depressive symptom in disabled middle-aged and elderly adults and to explore their predictors.Results:A total of 1 844 disabled middle-aged and elderly adults were analyzed for survey data. The trajectory of depressive symptoms in disabled middle-aged and elderly adults was categorized into four groups, including a low-level depression group, a depression remission group, a high-level depression group, and a depression worsening group. Logistic regression analyses revealed that gender, age, place of residence, chronic disease, self-rated health, social participation, cognition, and hearing ability were predictors of trajectories of depressive symptoms in disabled middle-aged and elderly adults.Conclusions:There is group heterogeneity in the trajectory of depressive symptoms among disabled middle-aged and elderly adults. Healthcare professionals can carry out targeted health management according to the characteristics of different categories of people.