[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.

Deactivation of the mitochondrial pyruvate dehydrogenase complex (PDC) is important for the metabolic switching of cancer cell from oxidative phosphorylation to aerobic glycolysis. Studies examining PDC activity regulation have mainly focused on the phosphorylation of pyruvate dehydrogenase (E1), leaving other post-translational modifications largely unexplored. Here, we demonstrate that the acetylation of Lys 488 of pyruvate dehydrogenase complex component X (PDHX) commonly occurs in hepatocellular carcinoma, disrupting PDC assembly and contributing to lactate-driven epigenetic control of gene expression. PDHX, an E3-binding protein in the PDC, is acetylated by the p300 at Lys 488, impeding the interaction between PDHX and dihydrolipoyl transacetylase (E2), thereby disrupting PDC assembly to inhibit its activation. PDC disruption results in the conversion of most glucose to lactate, contributing to the aerobic glycolysis and H3K56 lactylation-mediated gene expression, facilitating tumor progression. These findings highlight a previously unrecognized role of PDHX acetylation in regulating PDC assembly and activity, linking PDHX Lys 488 acetylation and histone lactylation during hepatocellular carcinoma progression and providing a potential biomarker and therapeutic target for further development.
Humans ; Acetylation ; Carcinoma, Hepatocellular/genetics* ; Liver Neoplasms/genetics* ; Pyruvate Dehydrogenase Complex/genetics* ; Gene Expression Regulation, Neoplastic ; Animals ; Mice ; Cell Line, Tumor ; Protein Processing, Post-Translational ; Histones/metabolism* ; Disease Progression

Objective To statistically analyze the perioperative results of patients with ABO-incompatible kidney trans-plantation(ABOi-KT),in order to explore the changes in blood group antibody of type-A/B recipients.Methods A total of 33 cases of blood group A/B ABOi-KT recipients in our hospital from January 2021 to October 2023 were recruited and divided into two groups of group A(n=18)and group B(n=15)according to the different blood types of recipient.The effects of preoperative plasmapheresis on antibody titer,antibody rebound and renal function after operation(serum urea ni-trogen,creatinine and estimated glomerular filtration rate on the 1st,3rd,7th and 14th day)were analyzed between the two groups.According to the postoperative rebound of blood type antibodies,33 recipients were divided into antibody rebound group(n=7)and non rebound group(n=26),and the differences in initial blood type antibody titers between the two groups were analyzed.Results There was no significant difference in the clearance rate of IgM with preoperative plasma ex-change between the two groups(Z=-0.26,P＞0.05);Levels of serum urea nitrogen and creatinine on the 1st,3rd,7th and 14th day after operation between group A and group B were not statistically significant(P＞0.05),the same as eGFR.Group B was more prone to rebound antibody compared with group A(P＜0.05).There was a significant difference in the in-itial IgM antibody titer between the blood type antibody rebound group and the non rebound group(Z=-2.127,P＜0.05),but no statistically significant difference in the initial IgG antibody titer(Z=-1.835,P＞0.05)between the two groups was found.Conclusion The patients type B receiving type AB kidney donors are more prone to rebound antibody after ABOi-KT operation compared to the the patients type A receiving type AB.

Objective:To investigate the molecular biological mechanism of Bw07 allele and its transferase alteration carried by a proband of ABw07 subtype.Methods:A 2-year-old male child was selected as the research object. The peripheral blood of the proband and his parents was identified for ABO blood type by the test tube method, and the ABO subgroup PCR-SSP detection and ABO gene sequencing were performed on the three individuals to determine their blood type genotypes. Finally, the effect of the p.Arg352Gln mutation on Bw07 transferase was verified by virtual mutation, DUET structure prediction, molecular dynamics analysis, and in vitro cellular experiments.Results:The serological phenotypes of the proband and his mother were ABw and Bw, respectively, while his father was normal A. The ABO subgroup PCR-SSP assay identified the three genotypes as Bw07/A, Bw07/O, and A/A, respectively.Sanger sequencing further verified that the proband and his mother carried the Bw07 gene, and virtual mutation showed that the intermolecular forces were weakened by the R352Q mutation. DUET predicted that this p.Arg352Gln mutation could affect the thermodynamic stability of Bw07 transferase. Molecular dynamics analysis confirmed that the alteration of thermodynamic stability was mainly related to the appearance of large fluctuations in the amino acid backbone atoms in the 125-133, 193-198 and 336-354 regions, and in vitro cellular experiments further verified the weakened antigen synthesis of Bw07 transferase.Conclusion:The formation of the ABw07 phenotype is associated with the mutation of the highly conserved Arg352 to Gln in Bw07 transferase.

Objective:To study the molecular basis for a proband with A subtype B of the ABO blood group and explore the influence of amino acid variant on the activity of glycosyltransferase (GT).Methods:A proband who had presented at the First Affiliated Hospital of Zhengzhou University on July 2, 2020 was selected as the study subject. Serological identification of the ABO blood groups of the proband and her family members were performed by gel card and test tube methods. The ABO gene of the proband was identified by PCR-sequence specific primers (PCR-SSP) and DNA sequencing. A 3D molecular homologous model was constructed to predict the impact of the variant on the stability of α-(1→3)-D-N-acetylgalactosamine transferase (GTA). Results:The red blood cells of the proband, her mother and two younger brothers showed weak agglutination with anti-A and strong agglutination with anti-B. The sera showed 1~2+ agglutination with Ac and no agglutination with Bc. Based on the serological characteristics, the proband was identified as AwB subtype. Pedigree analysis suggested that the variant was inherited from her mother. The blood group of the proband was identified as A223B type by PCR-SSP. ABO gene sequencing analysis showed that the proband has harbored heterozygous variants of c. 297A>G, c. 467C>T, c. 526C>G, c. 657C>T, c. 703G>A, c. 796C>A, c. 803G>C, c. 930G>A and c. 1055insA. Based on the results of clone sequencing, it was speculated that the genotype was ABO* A223/ ABO* B.01. There were c. 467C>T and c. 1055insA variants compared with ABO* A1.01, and c. 1055insA variant compared with ABO* A1.02. Homologous modeling showed that the C-terminal of A223 GT was significantly prolonged, and the local amino acids and hydrogen bond network have changed. Conclusion:Above results revealed the molecular genetics mechanism of A223B subtype. The c. 1055insA variant carried by the proband may affect the enzymatic activity of GTA and ultimately lead to weakening of A antigen.

Objective:To explore the RH genotype for a female with RhD(-) blood type and its molecular basis. Methods:A 26-year-old female who had attended the outpatient clinic of the First Affiliated Hospital of Zhengzhou University in August 2019 was selected as the study subject. Peripheral blood samples were collected from the proband and her parents for Rh phenotyping with gel card method. PCR-sequence-based typing (PCR-SBT) and DNA sequencing were used to determine the RHD zygosity and RH genotype of the proband and her parents. Homology modeling of Rh proteins was performed with bioinformatic software, and protein structural alterations caused by the variant was simulated by molecular dynamics. This study was approved by the Medical Ethics Committee of the First Affiliated Hospital of Zhengzhou University (Ethics No. 2023-KY-0870-003). Results:Serological tests showed that the proband and her father both had weakened e antigen of the Rh phenotype. PCR-SBT and DNA sequencing showed that the genotypes of the proband and her parents were dce/ dCE, dce/ DcE and dCE/ DcE, respectively. And the genotypes of the RHD and RHCE of the proband were RHD*01N.01/ RHD*01N.16, RHCE*01.01/RHCE*04, respectively. Protein simulation and molecular dynamics analysis revealed that the ce_16C variant resulted from RHCE* ce (c.48G>C) may alter the structure of intracellular and extracellular loops, mainly affecting the mobility of extracellular loops 2, 6 and intracellular loops 3, 4. Conclusion:Variant of the RHCE* ce allele c. 48G>C probably underlay the weakened e antigen in this proband.

Objective This study aimed to establish a pre-metastatic niche mouse model utilizing luciferase-labeled Lewis (Luc-Lewis) lung cancer cells and to assess the efficacy of this model employing both qualitative and quantitative methods. Methods C57BL/6 mice were categorized into two groups: a normal control group and a model group, each containing 15 individual mice. The pre-metastatic niche model was established via tail vein injection of Luc-Lewis lung cancer cells. Body mass were measured daily for all groups. Tumor fluorescence signals within the mice were detected using a high-throughput enzyme marker instrument. Lung tissue specimens were harvested to evaluate metastatic progression. HE staining was used to assess histopathological changes. Real-time quantitative PCR and Western blot analysis were used to detect the mRNA and protein expression of lysyl oxidase (LOX), matrix metalloproteinase 9 (MMP9), versican (VCAN), and fibronectin (FN), which are the specific markers for the formation of the microenvironment of lung tissues before metastasis. Results Significant declines in body mass and observable lethargy were noted in the model group when compared to the control group. Distinct fluorescence signals were observed in the lung tissue of the model group, demonstrating a positive correlation with the duration of model establishment. By day 14, elevated mRNA and protein expression levels of LOX, MMP9, VCAN, and FN were significantly evident. In addition, histopathological evaluations revealed augmented interstitial thickness, alveolar atrophy and significant inflammatory cell infiltration within the lung tissues of the model group. By the 21st day, metastatic lesions manifested in the lung tissues of the model group, suggesting an approximate pre-metastatic niche maturation timeline of 14 days. Conclusion A pre-metastatic niche mouse model for Lewis lung cancer is successfully established.
Mice ; Animals ; Lung Neoplasms/pathology* ; Matrix Metalloproteinase 9 ; Mice, Inbred C57BL ; Carcinoma, Lewis Lung ; Disease Models, Animal ; RNA, Messenger ; Tumor Microenvironment

Objective:To evaluate the effectiveness of esketamine during perioperative anesthesia for acute and chronic pain after cesarean section.Methods:One hundred and fifty patients scheduled for elective cesarean section under spinal anesthesia were randomly assigned into 2 equal groups ( n=75) using a random number table: esketamine group (group E) and control group (group C). Subarachnoid block was administered with 9-11 mg of hyperbaric bupivacaine with 0.33% glucose concentration. After the delivery of the fetus, 0.15 mg/kg (1 mg/ml) esketamine was pumped intravenously for 30 min in the group E, while the same dosage of normal saline was administered in the group C. Furthermore, patients received an intravenous patient controlled intravenous analgesia (PCIA) pump after surgery (100 μg sufentanil + 1.25 mg/kg esketamine + 8 mg ondansetron for the group E, 100 μg sufentanil + 8 mg ondansetron for the group C). Heart rate (HR), systolic blood pressure (SBP), and diastolic blood pressure (DBP) were recorded in the initial time of esketamine administration, and 5, 15 min, and 30 min after administration. The pain Numerical Rating Scale (NRS) score at rest and during coughing were recorded at 2, 6, 12, 24 h and 48 h after surgery. The first analgesic time and cumulative sufentanil consumption were recorded at 0-12 h, 12-24 h, 24-48 h, 0-24 h and 0-48 h after surgery. Moreover, we recorded the incidence of chronic pain at 3 and 6 months after surgery. Results:There were no significant differences in HR, SBP and DBP between the two groups immediately after administration of esketamine and 5, 15 min and 30 min after administration (all P>0.05). At rest or during coughing, the pain NRS score were significantly lower at 2, 6 h, and 12 h postoperatively in group E compared to group C (all P<0.05). The time to first analgesia in group E was significantly longer than the group C [(176.8±18.3)min vs (148.5±16.9)min, P<0.05]. The cumulative sufentanil consumption was significantly lower in group E during 0-12 h, 12-24 h, 0-24 h and 0-48 h postoperatively than in group C (all P<0.05), but there was no statistical significance between the two groups at 24-48 h ( P>0.05). There were no significant difference between the two groups in the incidence of chronic pain at 3 months and 6 months after surgery ( all P>0.05). The incidence of chronic pain in group E was lower than that in group C at 3 months [13.4%(9/67) vs 18.8%(13/69), P=0.392] and 6 months [10.7%(6/56) vs 16.1%(10/62), P=0.391], but the difference was not statistically significant. Conclusions:Perioperative administration of esketamine provided superior short-term analgesia after cesarean section and did not increase the psychotomimetic adverse event rate. However, the development of chronic pain was not restrained.

Objective:To explore the risk factors of perioperative outcomes of lung transplantation and establish a predictive model for delayed extubation after lung transplantation.Methods:From January 1, 2020 to December 31, 2022, 104 lung transplantation recipients were retrospectively collected to identify the risk factors of early post-operative outcome.According to the timing of extubation post-lung transplantation, they were assigned into two groups of normal(77 cases)and delayed(27 cases). Baseline profiles, type of primary diagnosis, cold ischemic duration and lung transplantation approach were compared between two groups.The factors with significant difference were examined by univariate and multivariate Logistic regression.Furthermore, multivariate logistic model was visualized by a nomogram.Receiver operating characteristic(ROC)curve and decision curve analysis(DCA) were performed for evaluating the model's predictive performance and its value for clinical utilization.Results:The postoperative mortality rate was 9.6%.Delayed extubation was a strong predictor for postoperative mortality.Cold ischemic time outperformed others variates in terms of delayed extubation prediction.AUC of cold ischemic time and multivariate logistic model was 0.75(95% CI: 0.69-0.81)and 0.87(95% CI: 0.82-0.91). Conclusions:Delayed postoperative extubation is a key predictor of early post-lung transplantation mortality.The established predictive model may effectively identify high-risk patients for preventive intervention and survival improvement post-lung transplantation.

Objective:To evaluate the clinical efficacy of biplane osteotomy in the treatment of malunion of Stephens-Sanders type Ⅱ calcaneal fracture.Methods:A retrospective study was conducted to analyze the clinical data of 31 patients who had been treated by biplane osteotomy at Sports Medicine Center, The First Affiliated Hospital of Army Medical University for malunion of Stephens-Sanders type Ⅱ calcaneal fracture from January 2019 to January 2022. There were 21 males and 10 females, with an age of (41.4±13.9) years and a duration from injury to diagnosis of (12.8±8.9) months. Functional and image scores were compared before surgery, 6 months after surgery, and at the last follow-up. Functional scores included the visual analogue scale (VAS) score, the American Orthopedic Foot and Ankle Society (AOFAS) score, and the pain interference (PI) and physical function (PF) indices in the Patient-Reported Outcomes Measurement Information System (PROMIS). Image scores included the Gissane angle, B?hler's angle, calcaneal pitch angle, length of the calcaneus, absolute foot height, and axial calcaneal width as measured on X-rays.Results:The operation time was (106.6±29.9) minutes for this cohort. All the 31 patients were followed up for (18.4±5.8) months. At 6 months after surgery and the last follow-up, the VAS scores [3 (2, 3), 2 (1, 3)], AOFAS scores [83 (76, 87), 85 (83, 87)], PI scores [(57±9), (48±6)], PF scores [53 (39, 61), 56 (54, 66)], Gissane angles (109.6°±14.1°, 109.3°±14.9°), B?hler angles (26.5°±11.6°, 26.9°±11.8°), calcaneal pitch angles [19.1° (14.5°, 23.9°), 19.9° (14.5°, 23.9°)], absolute foot heights [(76.5±9.6) mm, (76.0±9.9) mm], and axial calcaneal widths [(38.5±4.1) mm, (38.3±4.1) mm] were all significantly improved compared to the preoperative values [5 (4, 6), 62 (56, 67), (62±6), 47 (38, 51), 126.8°±13.1°, 11.8°±10.9°, 13.8° (8.2°, 18.7°), (71.0±9.1) mm, (42.8±5.5) mm] (all P<0.05). However, there was no statistically significant difference in the length of the calcaneus among pre-surgery, 6 months after surgery, and the last follow-up ( P>0.05). Conclusion:Biplane osteotomy is a surgical technique that demonstrates good clinical efficacy in the treatment of malunion of Stephens-Sanders type Ⅱ calcaneal fracture so that it should be promoted in clinic.