1.HOXD11 promotes the malignant biological behaviors of laryngeal squamous cell carcinoma through transcriptional regulation of Ki-67 activity
KONG Yannan ; LIU Liang ; LIU Yanli ; ZHAO Huiling ; NIU Yunfeng
Chinese Journal of Cancer Biotherapy 2026;33(4):379-388
[摘 要] 目的:探讨同源异型盒蛋白D11(HOXD11)对Ki-67的转录调控作用及其对喉鳞状细胞癌(LSCC)细胞恶性生物学行为的影响和机制。方法:结合高通量测序数据,通过GEO、UALCAN数据库分析HOXD11在LSCC中差异表达。收集2022年1月至2025年1月联勤保障部队第九八〇医院手术切除的60例LSCC患者的癌及癌旁组织标本,以及人LSCC细胞系AMC-HN-8、TU-177、TU-686和人正常喉上皮细胞(HNLEC),构建敲低或过表达HOXD11的细胞系,将细胞分为对照组、HOXD11敲低组及HOXD11过表达组。通过RT-qPCR法检测HOXD11和Ki-67基因在LSCC组织和细胞中mRNA表达水平,免疫组织化学(IHC)法分析两者在LSCC组织中的蛋白表达及分布,WB法进一步验证蛋白差异表达。采用MTS、克隆形成实验及Transwell实验分别检测敲低或过表达HOXD11对LSCC细胞增殖、迁移与侵袭的影响。通过双萤光素酶报告基因实验和染色质免疫沉淀(ChIP)实验验证HOXD11对Ki-67启动子活性的调控作用。结果:GEO和UALCAN数据库分析表明,HOXD11在LSCC中高表达(P < 0.01)。在LSCC组织中HOXD11和Ki-67 mRNA和蛋白表达均显著高于癌旁组织(均P < 0.01),同时,两者表达水平之间存在正相关(r = 0.26,P < 0.05);LSCC细胞系中HOXD11 mRNA表达显著高于HNLEC(均P < 0.01)。敲低HOXD11显著抑制LSCC细胞的增殖、迁移和侵袭能力(均P < 0.01),而过表达HOXD11则促进细胞的这些恶性生物学行为(均P < 0.01)。双萤光素酶报告基因实验及ChIP实验均证实,HOXD11可直接结合到Ki-67启动子区上,调控其表达(P < 0.01)。回复实验显示,过表达Ki-67可部分逆转敲低HOXD11对LSCC细胞增殖、迁移与侵袭的抑制作用(均P < 0.01)。结论:HOXD11在LSCC组织及细胞系中均呈高表达,其机制在于通过直接调控Ki-67的转录活性,从而增强LSCC细胞的增殖、迁移及侵袭能力。
2.Study on the construction of a red blood cell rare blood type database and physical repository in the Guangzhou Region
Zhijian LIAO ; Shuangshuang JIA ; Yuan SHAO ; Boquan HUANG ; Chunyan MO ; Jizhi WEN ; Runqing ZHANG ; Xia RONG ; Hong LUO ; Huaqin LIANG ; Yanli JI
Chinese Journal of Blood Transfusion 2026;39(5):619-628
Objective: To conduct screening for rare blood types within important blood group systems for the Chinese population, such as Rh, Duffy, Kidd, P1Pk, Diego, and MNS, in the Guangzhou region, and to establish a corresponding rare blood type database and physical repository. Methods: The saline medium microplate method was used to screen blood donors with the ccDEE phenotype combined with either Jk(a-) or Jk(b-). The polybrene microplate method was employed to screen for donors with Fy(a-), s(-), Lu(b-), Di(b-), k(-), and p phenotypes. The urea lysis microplate method was applied to screen for the Jk(a-b-) phenotype. A high-resolution melting (HRM) curve method was established for screening some donors with the Di(b-) phenotype. Subsequently, expanded phenotyping of antigens in the Rh, Kidd, MNS, Duffy, P1Pk, Lewis, Kell, and Lutheran blood group systems was performed on identified rare blood type donors using monoclonal antibodies. The test results are entered into the Rare Blood Type Bank Management System of the Guangzhou Blood Center, enabling functions such as confirmation reminders and cryopreservation storage when the donor donates again. Red blood cells of rare blood types are processed into frozen red blood cells for long-term storage. Results: Among voluntary blood donors, 16 cases of the ccDEE combined with Jk(a-) phenotype were identified (0.221 7%, 16/7 216); 10 cases of the ccDEE combined with Jk(b-) phenotype (0.138 6%, 10/7 216); 78 cases of the Fy(a-) phenotype (0.169 5%, 78/46 012); 39 cases of the Lu(b-) phenotype (0.138 2%, 39/28 214); 31 cases of the s(-) phenotype (0.081 8%, 31/37 913); 22 cases of the Di(b-) phenotype (0.029 9%, 22/73 691); 30 cases of the Jk(a-b-) phenotype (0.010 1%, 30/298 250); and 1 case of the k(-) phenotype (0.001 3%, 1/77 382), which was further identified as KELnull phenotype (K0). No p phenotype donors were identified (0/88 528). A total of 228 units of frozen red blood cells were prepared. The screening results were compared and analyzed with rare blood type data from other regions. Conclusion: This study, through a combination of different screening methods, significantly improved the efficiency of rare blood type screening while remaining cost-effective. By conducting large-scale screening and performing data informatization processing, a database and physical repository of rare blood types in the Guangzhou region were successfully established. This provides a strong guarantee for the timely supply of blood to patients with difficult-to-match and rare blood types in the region, effectively enhances the level of transfusion safety in the region, and offers a practical paradigm for constructing a comprehensive blood transfusion support system.
3.Mechanism of active ingredient compatibility of Dimocarpus longan Lour. leaves in improving glucose and lipid metabolism disorders in type 2 diabetic mellitus rats
Yanli LIANG ; Shijia AN ; Fengsheng LI ; Jiani MAI ; Anqi HUO ; Jiali WEI ; Zejuan ZHANG ; Shuyan QIN ; Wenqing HUANG ; Jie LIANG
China Pharmacy 2026;37(13):1697-1703
OBJECTIVE To explore the mechanism of the active ingredient compatibility(quercetin, quercitrin and kaempferol at a mass ratio of 2∶9∶3)of Dimocarpus longan Lour. leaves(abbreviated as CDL) on ameliorating glucose and lipid metabolism disorders in type 2 diabetes mellitus (T2DM) rats. METHODS SD rats were randomly divided into blank control group, model group, metformin hydrochloride group (100 mg/kg), and CDL high-, medium- and low-dose groups (280, 140, 75 mg/kg), with 10 rats in each group. Rats in the blank control group were fed with standard chow, while rats in the other groups were given high-sugar and high-fat diet combined with intraperitoneal injection of streptozotocin to establish the T2DM rat model. After successful modeling, rats in each administration group were given corresponding drug solution, and rats in the blank control group and model group were intragastrically administered with equal volume of pure water, once a day, for consecutive 4 weeks. Fasting blood glucose (FBG) was detected at fixed time every week. The curves of oral glucose tolerance test (OGTT) and intraperitoneal insulin tolerance test (IPITT) were plotted, and the area under curve (AUC) was calculated. The pancreatic islet function indexes [fasting insulin (FINS), homeostasis model assessment of insulin resistance (HOMA-IR), insulin sensitivity index (ISI)],blood lipid indexes [total cholesterol (TC), triglyceride (TG), low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol (HDL-C)] and hepatic glycogen content were determined. The pathological morphological changes of liver and pancreatic tissues were observed. The protein and mRNA expression levels of molecules related to phosphatidylinositol 3-kinase (PI3K)/protein kinase B (Akt) signaling pathway in liver tissues were detected. RESULTS Compared with the blank control group, the FBG, AUC of IPITT curve, AUC of OGTT curve, HOMA-IR, the levels of FINS, TC, TG and LDL-C, as well as the protein and mRNA expression of phosphatase and tensin homolog, forkhead box protein O1 and glycogen synthase kinase-3β in liver tissues were significantly increased in the model group ( P <0.05). ISI, the levels of HDL-C and hepatic glycogen content, along with the protein and mRNA expression of PI3K, insulin receptor substrate-1, Akt and protein expression of phosphorylated Akt in liver tissues were markedly decreased ( P <0.05). In model group rats, the arrangement of hepatocytes was irregular, the overall structure of pancreatic lobules was disordered, and a large number of inflammatory cell infiltration was observed. Compared with the model group, most of the above quantitative indexes were significantly reversed in the CDL high-dose group ( P <0.05), and the pathological lesions of liver and pancreas were obviously alleviated. CONCLUSIONS CDL can regulate glucose and lipid metabolism disorders, elevate insulin sensitivity and relieve insulin resistance in T2DM rats. Its mechanism may be related to the activation of the PI3K/Akt signaling pathway.
4.Mechanism of active ingredient compatibility of Dimocarpus longan Lour. leaves in improving glucose and lipid metabolism disorders in type 2 diabetic mellitus rats
Yanli LIANG ; Shijia AN ; Fengsheng LI ; Jiani MAI ; Anqi HUO ; Jiali WEI ; Zejuan ZHANG ; Shuyan QIN ; Wenqing HUANG ; Jie LIANG
China Pharmacy 2026;37(13):1697-1703
OBJECTIVE To explore the mechanism of the active ingredient compatibility(quercetin, quercitrin and kaempferol at a mass ratio of 2∶9∶3)of Dimocarpus longan Lour. leaves(abbreviated as CDL) on ameliorating glucose and lipid metabolism disorders in type 2 diabetes mellitus (T2DM) rats. METHODS SD rats were randomly divided into blank control group, model group, metformin hydrochloride group (100 mg/kg), and CDL high-, medium- and low-dose groups (280, 140, 75 mg/kg), with 10 rats in each group. Rats in the blank control group were fed with standard chow, while rats in the other groups were given high-sugar and high-fat diet combined with intraperitoneal injection of streptozotocin to establish the T2DM rat model. After successful modeling, rats in each administration group were given corresponding drug solution, and rats in the blank control group and model group were intragastrically administered with equal volume of pure water, once a day, for consecutive 4 weeks. Fasting blood glucose (FBG) was detected at fixed time every week. The curves of oral glucose tolerance test (OGTT) and intraperitoneal insulin tolerance test (IPITT) were plotted, and the area under curve (AUC) was calculated. The pancreatic islet function indexes [fasting insulin (FINS), homeostasis model assessment of insulin resistance (HOMA-IR), insulin sensitivity index (ISI)],blood lipid indexes [total cholesterol (TC), triglyceride (TG), low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol (HDL-C)] and hepatic glycogen content were determined. The pathological morphological changes of liver and pancreatic tissues were observed. The protein and mRNA expression levels of molecules related to phosphatidylinositol 3-kinase (PI3K)/protein kinase B (Akt) signaling pathway in liver tissues were detected. RESULTS Compared with the blank control group, the FBG, AUC of IPITT curve, AUC of OGTT curve, HOMA-IR, the levels of FINS, TC, TG and LDL-C, as well as the protein and mRNA expression of phosphatase and tensin homolog, forkhead box protein O1 and glycogen synthase kinase-3β in liver tissues were significantly increased in the model group ( P <0.05). ISI, the levels of HDL-C and hepatic glycogen content, along with the protein and mRNA expression of PI3K, insulin receptor substrate-1, Akt and protein expression of phosphorylated Akt in liver tissues were markedly decreased ( P <0.05). In model group rats, the arrangement of hepatocytes was irregular, the overall structure of pancreatic lobules was disordered, and a large number of inflammatory cell infiltration was observed. Compared with the model group, most of the above quantitative indexes were significantly reversed in the CDL high-dose group ( P <0.05), and the pathological lesions of liver and pancreas were obviously alleviated. CONCLUSIONS CDL can regulate glucose and lipid metabolism disorders, elevate insulin sensitivity and relieve insulin resistance in T2DM rats. Its mechanism may be related to the activation of the PI3K/Akt signaling pathway.
5.Analysis of clinical demand trends and supply status of RhD-negative red blood cell products
Weidong ZHANG ; Mingyue LIANG ; Yanli JI ; Ming WANG ; Yongju LIN ; Xia RONG
Chinese Journal of Blood Transfusion 2026;39(7):903-907
Objective: To retrospectively analyze the supply and demand of RhD-negative red blood cell (RBC) components, explore the dynamic changes in clinical demand and application trends of RhD-negative RBC products, and provide evidence for optimizing the preparation and supply management of various RhD-negative RBCs products. Methods: The total supply of RBCs and the demand and supply of RhD-negative RBC products from 2021 to 2025 were retrieved from the information management system. The trend chi-square test was performed using SPSS 22.0 statistical software to analyze the annual supply rate and repeat reservation rate of RhD-negative RBC products. Meanwhile, the clinical transfusion indications for RhD-negative RBC products in 2025 were further analyzed. Results: From 2021 to 2025, the supply rate of RhD-negative RBC products gradually increased from 0.42% (1 904.5 U/448 787 U) to 0.48% (1945U/402088.5U), showing an upward trend across years (P<0.05). The actual demand and supply of RhD-negative RBC products remained balanced, while the repeat reservation rate increased significantly from 29.73% (804 U/2 704 U) to 43.62% (1 505 U/3 450 U), showing an upward trend year by year (P<0.05). Fresh RhD-negative RBC products accounted for 87.09% (8 270 U/9 495.5 U) of the total supply, while RhD-negative frozen RBC products accounted for 12.91% (1 225.5 U/9 495.5 U). In 2025, therapeutic transfusion, elective surgery (including obstetric transfusion), and emergency transfusion accounted for 55.40% (1 077.5 U/1 945 U), 38.56% (750 U/1 945 U), and 6.04% (117.5 U/1 945 U) of the total supply of RhD-negative RBC products, respectively. RhD-negative frozen RBC products accounted for 67.23% (79 U/117.5 U) of emergency use and 29.47% (221 U/750 U) of elective surgery (including obstetrics) use; fresh RhD-negative RBC suspensions accounted for 93.92% (1 012 U/1 077.5 U) of therapeutic use and 70.56% (529 U/750 U) of elective surgery (including obstetrics) use. In recent years, RhD-negative washed RBC products and concentrated washed RBC products have been introduced as new products, and their clinical utilization has shown an increasing trend. Conclusion: Despite a decline in total blood collection volume, the security management system of RhD-negative RBC components in Guangzhou Blood Center has functioned largely meeting clinical demand. Advances in hospital technology had led to the routine application of RhD-negative concentrated washed RBCs. It is recommended that concentrated washed RBCs should be standardized with a formal definition and corresponding national quality standards should be established.
6.Identification of Jr(a-) rare blood type antibodies against anti-Jra: serological and molecular biology analysis and transfusion strategy
Yunxiang WU ; Hua WANG ; Ruiqing GUO ; Zhicheng LI ; Qing LI ; Dong XIANG ; Yanli JI ; Aijing LI ; Fengyong ZHAO ; Fei WANG ; Jiangtao ZUO ; Yi XU ; Yajun LIANG ; Demei ZHANG
Chinese Journal of Medical Genetics 2025;42(2):145-150
Objective:To report the blood group antigen and antibody specificity identification methods for a patient with high-frequency antibodies, and the process of finding and providing compatible blood for the patient.Methods:A patient sent from the Blood Transfusion Department of Shanxi Provincial People′s Hospital to Taiyuan Blood Center in November 2022 was selected for the study. Classical serological methods were used to determine the patient′s blood type, screen for unexpected antibodies, identify antibodies, and perform crossmatching. High-frequency antibody identification was carried out using red blood cells treated with various enzymes. Blood group genotyping was conducted using Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry (MALDI-TOF) and Sanger sequencing. Multiple strategies were employed to address the patient′s blood source problem. The study was approved by the Medical Ethics Committee of Taiyuan Blood Center [Ethics No. 2024 Ethics Review No.(2)].Results:①The patient′s blood type was B, RhD positive. Initial screening of the patient′s serum with multiple screening cells and antibody identification cells in saline medium was negative, but positive in antiglobulin medium. The patient′s serum showed varying reaction intensities with red blood cells treated with different enzymes. ②MALDI-TOF mass spectrometry and Sanger sequencing revealed a homozygous nonsense variant c. 376C>T (p.Gln126Ter) in the ABCG2 gene, resulting in the Jr(a-) phenotype. During family donor selection, the patient′s son was found to have a heterozygous variant c. 376C>T (p.Gln126Ter), and another heterozygous variant c. 421C>A (p.Gln141Lys), which predicted a Jr(a+ w) phenotype. ③Crossmatch tests confirmed the compatibility of blood from the patient′s son, which was used to address the urgent blood requirement. Later, rare blood from a Jr(a-) donor from the Guangzhou Blood Center was used for the patient′s ongoing treatment, saving the patient′s life. Conclusion:Combining classic serological testing with blood group gene typing techniques successfully identified the rare Jr(a-) blood type and high-frequency anti-Jra antibodies. Enzyme-treated red blood cell identification methods confirmed the presence of anti-Jra antibodies. By searching within the family and seeking help from other blood centers, compatible blood was found. This approach may provide insights for resolving similar complex blood matching problems in the future.
7.Identify the factors associated with treatment-free remission outcomes after imatinib discontinuation in children and adolescent patients with chronic myeloid leukemia
Huifang ZHAO ; Qian JIANG ; Weiming LI ; Yu ZHU ; Bingcheng LIU ; Qingshu ZENG ; Shuxia GUO ; Lixin LIANG ; Chunlei ZHANG ; Yingling ZU ; Yongping SONG ; Yanli ZHANG
Chinese Journal of Hematology 2025;46(9):800-805
Objective:To identify factors influencing treatment-free remission (TFR) outcomes in children and adolescent patients with chronic myeloid leukemia (CML) after imatinib (IM) discontinuation.Methods:This multicenter retrospective study analyzed 36 children and adolescent patients with CML from eight hematology centers in China (December 1, 2016, to September 27, 2024) who discontinued IM therapy with documented post-cessation outcomes. Clinical characteristics and molecular response dynamics were assessed. Univariate analysis and multivariate Cox proportional hazards regression models were employed to assess factors associated with TFR outcomes.Results:A total of 36 patients were documented, comprising 17 males and 19 females. The median ages at CML diagnosis and IM discontinuation were 11 years ( IQR: 5,16) and 20 years ( IQR: 14,25), respectively. The median time from IM initiation to first deep molecular response (DMR) was 21 months ( IQR: 13, 38). Pre-discontinuation, patients received IM for a median duration of 96 months ( IQR: 84, 121) and maintained DMR for 74 months ( IQR: 63, 89). With a median post-discontinuation follow-up of 38 months ( IQR: 15, 68), cumulative TFR rates at 6, 12, 24, and 36 months were 74.1%, 60.7%, 60.7%, and 56.0%, respectively, generating an overall TFR rate of 58.3%. Fifteen patients lost major molecular response at a median of 5 months post-discontinuation ( IQR: 3, 11). All 15 patients resumed tyrosine kinase inhibitor therapy, comprising 13 who restarted IM and 2 who switched to dasatinib. By the last follow-up, 13 (86.7% ) patients regained DMR after a median treatment duration of 5 months ( IQR: 3, 17), and no disease progression occurred in any patient. Withdrawal syndrome occurred in 2 (5.6% ) patients. Univariate analysis revealed significantly higher TFR rates in patients with pre-discontinuation IM duration of ≥100 months vs <100 months (82.4% vs 36.8%, P=0.017) and pre-discontinuation DMR duration of ≥72 months vs <72 months (84.2% vs 29.4%, P=0.003). Multivariate Cox analysis identified pre-discontinuation DMR duration as an independent protective factor for TFR ( HR=5.419, 95% CI: 1.524–19.272, P=0.009) . Conclusion:DMR duration was identified as an independent protective factor influencing TFR outcomes in children and adolescent patients with CML after IM discontinuation. Patients who maintained DMR for ≥72 months before IM discontinuation demonstrated a significantly higher TFR rate.
8.Correlation analysis of smoking behavior and cardiovascular risk factors among employees in a heavy industry enterprise
Qian ZHANG ; Xi WANG ; Jin WANG ; Yanli LIANG ; Yubo ZHENG ; Rui TANG ; Juanjuan JIN ; Yanping REN
Chinese Journal of Health Management 2025;19(10):781-786
Objective:To explore the correlation between smoking behavior of employees in heavy industry enterprises and risk factors for cardiovascular and cerebrovascular diseases.Methods:A cross-sectional study design was adopted. A questionnaire survey and physical examination data collection were conducted on employees of a heavy industry enterprise in Shaanxi Province using a cluster sampling method from March to April 2024. A total of 2 209 cases met the inclusion criteria. According to the smoking index (SI), participants were divided into a non-smoking group (SI=0, 1 316 cases), a low smoking group (SI400, 656 cases), and a high smoking group (SI≥400, 237 cases). According to the fagerstrom test for nicotine dependence (FTND) score, participants were divided into a non-dependence group (FTND=0, 1 316 cases), a mild dependence group (FTND=1-3, 623 cases), a moderate dependence group (FTND=4-6, 204 cases), and a severe dependence group (FTND≥7, 66 cases). Spearman rank correlation, univariate, and multivariate logistic regression analyses were used to compare differences in blood pressure, fasting blood glucose, total cholesterol, low-density lipoprotein cholesterol, high-density lipoprotein cholesterol, triglycerides, uric acid, homocysteine, and body mass index among the groups.Results:As the SI and FTND scores increased, blood pressure, low-density lipoprotein cholesterol, triglycerides, uric acid, and homocysteine all showed an increasing trend, while high-density lipoprotein cholesterol showed a decreasing trend (all P0.05). The systolic blood pressure, diastolic blood pressure, fasting blood glucose, low density lipoprotein cholesterol (LDL-C), triglycerides, serum uric acid, and homocysteine in the high smoking group were higher than those in the non-smoking group [(124.93±16.55) vs (122.32±16.62) mmHg (1 mmHg=0.133 kPa), (80.86±11.10) vs (76.18±11.63) mmHg, 5.58 vs 5.29 mmol/L, (3.13±0.65) vs (2.95±0.56) mmol/L, 1.70 vs 1.09 mmol/L, (336.80±69.94) vs (299.00±86.43) μmol/L, 14.20 vs 11.76 μmol/L, all P0.001], the high-smoking group had lower HDL cholesterol than the non-smoking group [(1.34±0.30) vs (1.39±0.27) mmol/L, P0.001], the systolic blood pressure, diastolic blood pressure, fasting blood glucose, total cholesterol, low-density lipoprotein cholesterol, triglycerides, serum uric acid, and homocysteine in the severely dependent group were higher than those in the non-dependent group [(132.09±16.69 )vs (122.32±16.62) mmHg, (81.32±12.97) vs (76.18±11.63) mmHg, 5.61 vs 5.29 mmol/L, (4.98±0.91 )vs (4.70±0.88) mmol/L, (3.19±0.62) vs (2.95±0.56) mmol/L, 1.87 vs 1.09 mmol/L, (328.95±75.21) vs (299.03±86.43) μmol/L, 14.38 vs 11.76 μmol/L, all P0.05], and the high density lipoprotein cholesterol (HDL-C) in the severely dependent group was lower than that in the non-dependent group [(1.32±0.30) vs (1.39±0.27) mmol/L, P0.001]. Multivariate analysis showed that SI and degree of tobacco dependence (DTD), were significantly correlated with fasting blood glucose, triglycerides, and LDL cholesterol, but the effect size of SI was too small ( OR=1.001, 95% CI: 1.000-1.001, P=0.002, OR=1.001, 95% CI: 1.001-1.002, P0.001, OR=1.001, 95% CI: 1.000-1.001, P=0.032). Conclusions:Smoking behavior among employees in heavy industry enterprises is significantly correlated with risk factors for cardiovascular and cerebrovascular diseases. The degree of tobacco dependence is closely related to the risk of abnormal fasting blood glucose, low-density lipoprotein cholesterol, and triglycerides.
9.COPB1 promotes the development and progression of esophageal squamous cell carcinoma by activating the PI3K/AKT pathway and regulating the tumor immune microenvironment
LIN Yan ; YU Shuangjian ; JIA Sifan ; LI Feiyu ; ZHAO Chenpu ; DONG Zhiming ; SHEN Supeng ; LIANG Jia ; GUO Yanli
Chinese Journal of Cancer Biotherapy 2025;32(12):1236-1246
[摘 要] 目的:探究包被蛋白复合体β1亚基(COPB1)在食管鳞状细胞癌(ESCC)中的表达,及其对ESCC细胞恶性生物学行为的影响、作用机制及临床意义。方法:采用2014~2018年间在河北医科大学第四医院生物样本库中82例ESCC组织及癌旁组织,常规培养正常食管鳞状上皮细胞HEEC和食管癌细胞KYSE-150、KYSE-170、Eca109、TE1、KYSE-30、KYSE-450,用转染试剂将pcDNA3.1-vector(空载体)、pcDNA3.1-COPB1载体,si-NC和si-COPB1转染至KYSE-150、TE1细胞中,记为NC、COPB1-OE、si-NC和si-COPB1组。用数据库数据分析COPB1 mRNA在泛癌组织中的表达及其表达与免疫细胞浸润的关系,qPCR法检测ESCC组织和细胞中COPB1、PIK3CB、CD68、CD163、CD206、ARG1、IL-10 mRNA水平表达情况,WB法检测ESCC组织和各组细胞中的COPB1、PI3K、CD68、CD163、CD206、p-AKT蛋白表达,克隆形成实验和MTS实验检测各组细胞的增殖能力,划痕愈合实验和Transwell实验检测各组细胞的迁移和侵袭能力,免疫组织化学染色(IHC)法检测ESCC组织中COPB1和CD206蛋白表达。以人单核细胞白血病细胞(THP-1)构建巨噬细胞模型,用佛波酯(PMA)和IL-3和IL-4和ESCC细胞上清液诱导巨噬细胞转型,用qPCR和WB法检测CD68和CD206m RNA和蛋白的表达。结果:COPB1在泛癌组织和ESCC组织中均呈高表达且与淋巴结转移和TNM分期有关联(均P < 0.01),COPB1高表达的ESCC患者总生存期短(P < 0.05),COPB1是潜在的ESCC的诊断标志物。COPB1在KYSE-150和TE1细胞中也呈高表达(均P < 0.05),过表达或敲减COPB1可明显抑制或促进KYSE-150和TE1细胞的增殖能力、迁移和侵袭能力(均P < 0.05)。COPB1表达变化诱导的差异表达基因主要富集于PI3K/AKT通路(均P < 0.001), COPB1可促进PI3K/AKT通路的活化(P < 0.05),COPB1高表达可导致M2型巨噬细胞浸润增加(P < 0.05),COPB1高表达促进TAM/M2极化(P < 0.05)。结论:COPB1在ESCC组织中呈高表达,其可激活PI3K/AKT通路及调控肿瘤免疫微环境促进 ESCC发生发展,COPB1有望成为ESCC诊断和预后的生物标志物及治疗靶点。
10.Identification of Jr(a-) rare blood type antibodies against anti-Jra: serological and molecular biology analysis and transfusion strategy.
Yunxiang WU ; Hua WANG ; Ruiqing GUO ; Zhicheng LI ; Qing LI ; Dong XIANG ; Yanli JI ; Aijing LI ; Fengyong ZHAO ; Fei WANG ; Jiangtao ZUO ; Yi XU ; Yajun LIANG ; Demei ZHANG
Chinese Journal of Medical Genetics 2025;42(2):145-150
OBJECTIVE:
To report the blood group antigen and antibody specificity identification methods for a patient with high-frequency antibodies, and the process of finding and providing compatible blood for the patient.
METHODS:
A patient sent from the Blood Transfusion Department of Shanxi Provincial People's Hospital to Blood Transfusion Technology Research Laboratory of Taiyuan Blood Center in November 2022 was selected for the study. Classical serological methods were used to determine the patient's blood type, screen for unexpected antibodies, identify antibodies, and perform crossmatching. High-frequency antibody identification was carried out using red blood cells treated with various enzymes. Blood group genotyping was conducted using Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry (MALDI-TOF) and Sanger sequencing. Multiple strategies were employed to address the patient's blood source problem. The study was approved by the Medical Ethics Committee of Taiyuan Blood Center [Ethics No. 2024 Ethics Review No.(2)].
RESULTS:
The patient's blood type was B, RhD positive. Initial screening of the patient's serum with multiple screening cells and antibody identification cells in saline medium was negative, but positive in antiglobulin medium. The patient's serum showed varying reaction intensities with red blood cells treated with different enzymes. MALDI-TOF mass spectrometry and Sanger sequencing revealed a homozygous nonsense variant c.376C>T (p.Gln126Ter) in the ABCG2 gene, resulting in the Jr(a-) phenotype. During family donor selection, the patient's son was found to have a heterozygous variant c.376C>T (p.Gln126Ter), and another heterozygous variant c.421C>A (p.Gln141Lys), which predicted a Jr(a+w) phenotype. Crossmatch tests confirmed the compatibility of blood from the patient's son, which was used to address the urgent blood requirement. Later, rare blood from a Jr(a-) donor from the Guangzhou Blood Center was used for the patient's ongoing treatment, saving the patient's life.
CONCLUSION
Combining classic serological testing with blood group gene typing techniques successfully identified the rare Jr(a-) blood type and high-frequency anti-Jra antibodies. Enzyme-treated red blood cell identification methods confirmed the presence of anti-Jra antibodies. By searching within the family and seeking help from other blood centers, compatible blood was found. This approach may provide insights for resolving similar complex blood matching problems in the future.
Humans
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Blood Grouping and Crossmatching/methods*
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Blood Group Antigens/immunology*
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Blood Transfusion
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Male
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Isoantibodies/blood*
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Female
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Genotype

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