1.Therapeutic effect of PD-1 monoclonal antibody combined with cisplatin or gemcitabine chemotherapy in a mouse xenograft model with KRAS mutant non-small cell lung cancer A549 cells
LI Xiongbing ; ZHOU Ruifen ; LI Jiali ; WANG Hanjiao ; WANG Chao ; LI Jing ; CAO Zhe ; SHU Chengrong
Chinese Journal of Cancer Biotherapy 2024;31(4):371-376
[摘 要] 目的:探讨程序性死亡受体-1(PD-1)单抗联合顺铂或吉西他滨在KRAS基因突变非小细胞肺癌(NSCLC)A549细胞移植瘤小鼠模型治疗中的作用。方法:构建免疫系统-肿瘤双人源化A549细胞小鼠移植瘤模型,将60只小鼠按随机数字表法分成6组(10只/组),分别为对照组(200 μL/kg PBS)、PD-1单抗组(20 mg/kg PD-1单抗)、顺铂组(3 mg/kg顺铂)、PD-1单抗+顺铂组(20 mg/kg PD-1单抗+3 mg/kg顺铂)、吉西他滨组(30 mg/kg吉西他滨)和PD-1单抗+吉西他滨组(20 mg/kg PD-1单抗+30 mg/kg吉西他滨)。TUNEL和DAPI双染色法检测移植瘤组织中细胞凋亡水平,测量移植瘤体积和质量并计算肿瘤生长抑制率,免疫组化法检测移植瘤微血管密度(MVD)。结果:成功构建免疫系统-肿瘤双人源化NSCLC A549细胞小鼠移植瘤模型,PD-1单抗+顺铂组移植瘤的细胞凋亡率、肿瘤生长抑制率均最高,移植瘤体积、质量和MVD均最小,与其他5组小鼠比较差异均有统计学意义(均P<0.05)。结论:顺铂与PD-1单抗具有协同活性,而吉西他滨拮抗PD-1单抗的治疗作用。提示PD-1单抗联合顺铂对KRAS突变NSCLC A549细胞移植瘤小鼠的疗效更好。
2.Erratum to "Tanshinone IIA Protects Endothelial Cells from H2O2 -Induced Injuries via PXR Activation" Biomol Ther 25(6), 599-608 (2017)
Haiyan ZHU ; Zhiwu CHEN ; Zengchun MA ; Hongling TAN ; Chengrong XIAO ; Xianglin TANG ; Boli ZHANG ; Yuguang WANG ; Yue GAO
Biomolecules & Therapeutics 2024;32(2):261-261
3.Construction of a nomogram prediction model for survival prognosis of patients with advanced non-small cell lung cancer after PD-1 inhibitor treatment
Xiongbing LI ; Ruifen ZHOU ; Jiali LI ; Hanjiao WANG ; Chao WANG ; Jing LI ; Zhe CAO ; Chengrong SHU
International Journal of Laboratory Medicine 2023;44(24):2975-2979
Objective To explore the effect of neutrophil to lymphocyte ratio(NLR)and other related in-dicators on the prognosis of advanced non-small cell lung cancer patients treated with programmed death 1(PD-1)inhibitor and construct a nomogram prediction model.Methods A total of 198 patients with advanced non-small cell lung cancer who received PD-1 inhibitor treatment in the hospital from February 2020 to April 2022 were selected and followed up to August 2022.According to the clinical outcome,they were divided into the death group(46 cases)and the survival group(152 cases).The clinical data of the death group and the survival group were recorded,and the prognostic factors of advanced non-small cell lung cancer patients trea-ted with PD-1 inhibitor were analyzed.Receiver operating characteristic(ROC)curve was used to analyze the predictive value of NLR,platelet to lymphocyte ratio(PLR)and lymphocyte to monocyte ratio(LMR)for the prognosis of patients.Multivariate Logistic regression model was used to analyze the independent risk factors affecting the prognosis of patients.A prediction nomogram model for the prognosis of patients was construc-ted using R software 4.0"rms"package,and the calibration curve was used to internally validate the nomo-gram prediction model.Results Compared with the survival group,the proportion of smoking,TNM stageⅣ,ECOG score 2,and NLR,PLR,LMR were higher(P<0.05).The area under the curve of NLR,PLR and LMR were 0.707,0.793 and 0.819,respectively,and the optimal cut-off value were 4.72%,179.21%and 3.44%,respectively.Smoking,TNM stage,ECOG score,NLR,PLR,and LMR were independent risk factors for the prognosis of advanced non-small cell lung cancer patients treated with PD-1 inhibitor(P<0.05).The internal validation results show that the nomogram inhibitor treatment of PD-1 model prediction the prognosis of patients with advanced non-small cell lung cancer C-index was 0.847(95%CI 0.769-0.902),the calibra-tion curve tends to be the ideal curve.The threshold value of the nomogram model for predicting the prognosis of patients with advanced non-small cell lung cancer treated with PD-1 inhibitor was>0.22.The nomogram prediction model provided a net clinical benefit,and the net clinical benefit was higher than that of smoking,TNM stage,ECOG score,NLR,PLR and LMR.Conclusion Based on smoking,TNM stage,ECOG score,NLR,PLR,and LMR,a nomogram prediction model for the prognosis of advanced non-small cell lung cancer patients treated with PD-1 inhibitor is constructed,which has important clinical application value.
4.Changes and significance of granulocyte-like myeloid-derived suppressor cells during acute phase of Kawasaki disease
Pengqiang WEN ; Guobing WANG ; Jiehua MEI ; Zhongxiang QI ; Li YANG ; Mingguo XU ; Cong LIU ; Chengrong LI
Chinese Journal of Microbiology and Immunology 2022;42(7):540-548
Objective:To investigate the changes and significance of granulocyte-like myeloid-derived suppressor cells (G-MDSC) in the acute phage of Kawasaki disease (KD).Methods:Forty-two children with acute KD were enrolled in the present study and 32 age-matched healthy children were selected as control group. The proportion of HLA-DR -CD11b + CD33 + CD14 -CD15 + G-MDSC, the concentration of reactive oxygen species (ROS) and the expression of arginase-1 (Arg-1), programmed death-ligand 1 (PD-L1), cytotoxic T lymphocyte associated protein 4 (CTLA4), glycoprotein 130 (gp130) and phosphorylated signal transducer and activator of transcription 3 (pSTAT3) at protein level were detected by flow cytometry. Quantitative real-time PCR was used to measure the expression of inducible nitric oxide synthase (iNOS), interferon regulatory factor 8 (IRF-8), IL-6 receptor α subunit (IL-6Rα), granulocyte colony-stimulating factor receptor (G-CSFR), CCAAT/enhancer binding protein β (C/EBPβ), suppressor of cytokine signaling 1 (SOCS1) and SOCS3 at mRNA level in G-MDSC. Chromatin immunoprecipitation was performed to detect the acetylation of histone H3 at the promoters of SOCS1 and SOCS3 genes. Plasma concentrations of IL-6 and granulocyte colony-stimulating factor (G-CSF) and protein levels of IL-10, transforming growth factor-β (TGF-β) and nitric oxide (NO) in the culture supernatant of G-MDSC stimulated with LPS were measured by ELISA. Results:(1) Compared with the control group, the proportion of HLA-DR -CD11b + CD33 + CD14 -CD15 + G-MDSC as well as the concentration of ROS and the expression of inhibitory molecules (Arg-1, PD-L1 and CTLA4) in G-MDSC increased significantly in patients with acute KD ( P<0.05). Moreover, the concentrations of IL-10 and TGF-β in culture supernatant of G-MDSC were also higher than those of the control group after stimulation with lipopolysaccharide for 48 h ( P<0.05). All of the seven afore-mentioned indexes in KD patients with coronary artery lesion (CAL group ) were lower than those in patients without coronary artery lesion (NCAL group) ( P<0.05), and restored to some extent after IVIG therapy ( P<0.05). There were no statistical differences in iNOS expression or NO concentration in culture supernatant of G-MDSC among different groups ( P<0.05). (2) Plasma concentrations of IL-6 and G-CSF, and the expression of IL-6Rα, gp130, G-CSFR, pSTAT3 and C/EBPβ increased remarkably during acute phase of KD ( P<0.05). The expression of IRF-8 at transcription level in patients with acute KD was found to be lower than that of healthy controls ( P<0.05), and restored significantly after IVIG therapy ( P<0.05). Moreover, the plasma concentrations of IL-6 and G-CSF and the expression of IL-6Rα, gp130, G-CSFR and IRF-8 in the CAL group were higher than those in the NCAL group ( P<0.05), while the expression of pSTAT3 and C/EBPβ was lower in the CAL group ( P<0.05), which were restored by IVIG therapy ( P<0.05). (3) In patients with acute KD, the expression of SOCS1 and SOCS3 at mRNA level and histone acetylation at the promoters of SOCS1 and SOCS3 genes were reduced significantly in comparison with those in healthy controls ( P<0.05) , but were increased remarkably after IVIG treatment( P<0.05). The four indexes were higher in the CAL group than in the NCAL group ( P<0.05). Pearson correlation analysis showed the expression of SOCS1 and SOCS3 was negatively correlated with the protein level of pSTAT3 in G-MDSC of patients with acute KD ( r=-0.46 and -0.32, P<0.05). Conclusions:Changes in the number and function of G-MDSC caused by aberrant histone acetylation at SOCS1 and SOCS3 genes might contribute to the immune dysfunction and vascular damage in patients with KD.
5.Changes and significance of CD8 + CD28 - regulatory T cells in acute phase of Kawasaki disease
Chunxiu LYU ; Yuxin GUO ; Pengqiang WEN ; Mingguo XU ; Guobing WANG ; Zhe SU ; Cong LIU ; Chengrong LI
Chinese Journal of Microbiology and Immunology 2022;42(10):791-797
Objective:To investigate the changes of CD8 + CD28 - regulatory T cells (Treg) and its role in the pathogenesis of Kawasaki disease (KD). Methods:A total of 48 children with KD were enrolled in the present study from June 2019 to December 2021. Blood samples were collected from them during acute phage of KD and after intravenous immunoglobulin (IVIG) treatment. Another 32 age-matched healthy children were recruited as control group. The proportions of CD8 + CD28 -Treg cells and the expression of programmed cell death protein 1 (PD-1), factor associated suicide ligand (FasL), inducible T-cell co-stimulator ligand (ICOSL), CD80 and CD86 protein were evaluated by flow cytometry. The expression of Helios, perforin, granzyme B, immunoglobulin-like transcript 3 (ILT3) and ILT4 at the transcription level was measured by real-time PCR. Concentrations of IL-10 and TGF-β in the culture supernatants of CD8 + CD28 -Treg cells stimulated with activated CD4 + T cells were measured by ELISA. Results:⑴ The proportions of CD8 + CD28 -Treg cells and the expression of Helios in patients with acute KD were higher than those in the control group ( P<0.05), and reduced remarkably after IVIG therapy ( P<0.05). The two afore-mentioned indexes were lower in patients combined with coronary artery lesion (CAL) than in those without coronary artery lesion (NCAL) ( P<0.05). ⑵ Compared with the control group, the patients with acute KD showed increased expression of FasL, PD-1, ICOSL and perforin in CD8 + CD28 -Treg cells ( P<0.05). The concentrations of IL-10 and TGF-β1 in the culture supernatants of CD8 + CD28 -Treg cells from patients with acute KD were lower than those in the control group after stimulation with activated CD4 + T cells ( P<0.05), which restored to some extent after IVIG treatment ( P<0.05). All of the six above-mentioned indexes in the CAL group were found to be lower than those in the NCAL group ( P<0.05). There were slight differences in granzyme B expression between different groups ( P>0.05). (3) In comparison with the healthy controls, the patients with acute KD showed overexpressed co-stimulatory molecules such as CD80 and CD86 on CD14 + cells ( P<0.05) and up-regulated expression of inhibitory molecules ILT3 and ILT4 ( P<0.05), which were restored remarkably after IVIG treatment ( P<0.05). Furthermore, the expression of CD80 and CD86 at protein level increased in the CAL group than in the NCAL group ( P<0.05), while the expression of ILT3 and ILT4 at transcriptional level decreased in the CAL group ( P<0.05). Conclusions:Relative insufficiency and impaired function of CD8 + CD28 -Treg cells might be one of the important factors resulting in immune dysfunction and vascular damage in KD patients.
6.Activating Effect of Effective Components in Medicinal and Edible Substances on Human Pregnane X Receptor and Cytotoxicity Screening
Zuqi ZHANG ; Guangchen ZHANG ; Panpan RUAN ; Yi LIN ; Hongling TAN ; Chengrong XIAO ; Zengchun MA ; Yuguang WANG ; Yue GAO
Chinese Journal of Experimental Traditional Medical Formulae 2022;28(13):42-51
ObjectiveTo explore the activating effects of ten important effective components in seven medicinal and edible substances on human pregnane X receptor (PXR), including Glycyrrhizae Radix et Rhizoma (liquiritin and glycyrrhizic acid), Houttuyniae Herba (quercetin and houttuyfonate), Prunellae Spica (rosmarinic acid), Cassiae Semen (aurantio-obtusin), Poria (pachymic acid), Lilii Bulbus (Lilium brownii saponin and colchicine), and Lycii Fructus (Lycium barbarum polysaccharide) and screen potentially toxic components. MethodCell counting kit-8 (CCK-8) assay was used to investigate the cytotoxic effect of liquiritin, glycyrrhizic acid, quercetin, houttuyfonate, rosmarinic acid, pachymic acid, aurantio-obtusin, and colchicine (10, 20, and 50 μmol·L-1), and L. brownii saponin and L. barbarum polysaccharide (10, 20, and 50 mg·L-1) on normal human hepatocyte cell line (L02). The release of lactate dehydrogenase (LDH) in L02 cells after drug treatments was detected by the biochemical analyzer. The apoptosis induced by ten effective components was explored by Hoechst 33342 staining. The secreted luciferase reporter system was used to co-transfect the PXR expression vector and reporter gene vector containing cytochrome P450 3A4 (CYP3A4) transcriptional regulatory region into L02 cells, with 10 μmol·L-1 rifampicin (RIF) as a positive control. After treated with liquiritin, glycyrrhizic acid, quercetin, houttuyfonate, rosmarinic acid, aurantio-obtusin, pachymic acid, and colchicine (5, 10, and 20 μmol·L-1) and L. brownii saponin and L. barbarum polysaccharide (5, 10, and 20 mg·L-1) for 24 h, the cells were tested for secreted luciferase activity. ResultCompared with the control group, colchicine, L. brownii saponin, and quercetin decreased the cell viability (P<0.05, P<0.01). Compared with the control group, quercetin, rosmarinic acid, glycyrrhizic acid, colchicine, aurantio-obtusin, and pachymic acid increased the release rate of LDH in L02 cells (P<0.05, P<0.01). The proportion of hyperchromatic nuclei increased gradually after rosmarinic acid, liquiritin, and L. barbarum polysaccharide treatments as compared with the control group (P<0.05, P<0.01). In terms of co-transfection of pcDNA3.1-PXR and pGLuc-CYP3A4 into L02 cells, compared with the control group, aurantio-obtusin and pachymic acid showed activating effects on PXR (P<0.05), whereas liquiritin and glycyrrhizic acid showed inhibitory effects (P<0.05). ConclusionThe findings suggest that when medicinal and edible substances are taken for a long time, attention should be paid to their influence on drug-metabolizing enzymes and possible interactions, so as to improve their safety.
7.Effects of Notch1 signaling on regulatory T cells and coronary artery lesions in childhood Kawasaki disease
Yuxin GUO ; Li YANG ; Guobing WANG ; Pengqiang WEN ; Zhongxiang QI ; Mingguo XU ; Cong LIU ; Chengrong LI
Chinese Journal of Rheumatology 2022;26(10):649-656,C10-1
Objective:To explore the effect of Notch1 signaling on regulatory T cells and its roles in vascular damage in patients with Kawasaki disease (KD).Methods:A total of 42 children with KD were enrolled in the present study from March 2019 to June 2020, as 32 age-matched healthy children were recruited as control. The proportions of CD4 +CD25 hiFoxp 3+ regulatory T cells (Treg) and expressions of transcription factor forkhead box protein 3 (Foxp3), cytotoxic T lymphocyte associated antigen-4 (CTLA4), glucocorticoid-induced tumor necrosis factor receptor (GITR), and Notch1 protein were evaluated by flow cytometry. Chromatin immunoprecipitation was conducted to detect acetylation level of histone H4 (H4Ac) associated with the promoter of Foxp3 gene and its binding abilities of Notch1 intracellular domain 1 (NICD1), recombination signal binding protein for immunoglobulin kappa J region (RBP-J) and p300 in CD4 + T cells. Transcription levels of Foxp3, presenilin 1 (PSEN1), mastermind like transcriptional coactivator 1 (MAML1), and RBP-J in CD4 + T cells were determined by real-time polymerase chain reaction (PCR). Concentrations of interleukin (IL)-10 and transforming growth factor-β (TGF-β) in plasma and culture supernatant stimulated with Jagged1 were measured by enzyme linked immunosorbent assay. Independent-sample t-test, Pearson correlation analysis was used as the statistical method in this study. Results:① The frequencies of Treg in acute KD patients decreased significantly [(4.3±1.5)% vs (7.9±2.9)%; t=6.41, P<0.001], as protein levels of Foxp3, CTLA4 and GITR and concentrations of IL-10 and TGF-β in plasma reduced remarkably in acute KD patients ( t=6.87, P<0.001; t=4.26, P<0.001; t=7.88, P<0.001; t=8.42, P<0.001; t=13.01, P<0.001). All parameters afore-mentioned in patients combined with coronary artery lesions (CAL) were lower than those of patients without coronary artery lesions (NCAL) ( t=5.83, P<0.001; t=3.83, P<0.001; t=3.28, P=0.002; t=5.05, P<0.001; t=5.96, P<0.001; t=5.17, P<0.001), and increased after therapy ( t=7.13, P<0.001; t=6.10, P<0.001; t=4.31, P<0.001; t=6.55, P<0.001; t=7.40, P<0.001; t=7.84, P<0.001). ② H4Ac associated with promoter of Foxp3 gene and the binding abilities of NICD1 and p300 in acute KD patients were lower than those of the controls ( t=10.25, P<0.001; t=6.93, P<0.001; t=6.75, P<0.001), and increased remarkably after therapy ( t=7.72, P<0.001; t=4.16, P<0.001; t=5.76, P<0.001). Meanwhile, the three items in CAL group were found to be less than those of NCAL group ( t=6.08, P<0.001; t=2.66, P=0.011; t=6.02, P<0.001). Pearson correlation analysis showed a positive correlation between H4Ac associated with Foxp3 promoter and its mRNA level in acute KD patients ( r=0.47, P<0.001). No statistical significant difference about the binding ability of RBP-J with Foxp3 promoter were found among the groups ( t=0.57, P>0.05; t=0.61, P>0.05; t=1.20, P>0.05). ③ Protein level of Notch1 and the expressions of PSEN1, MAML1 and RBP-J mRNA in CD4 + T cells from acute KD patients were down-regulated remarkably ( t=5.28, P<0.001; t=6.31, P<0.001; t=11.78, P<0.001; t=8.06, P<0.001), and restored after therapy ( t=4.77, P<0.001; t=6.43, P<0.001; t=11.95, P<0.001; t=7.79, P<0.001). In parallel, the four indexes aforementioned of CAL group were lower than those of NCAL group ( t=3.16, P=0.003; t=4.13, P<0.001; t=5.42, P<0.001; t=4.05, P<0.001). Upon rhJagged1 stimulation for 48 hours, H4Ac level of Foxp3 promoter and its binding abilities with NICD1 and p300 in CD4 + T cells in KD patients and control group was significantly higher than those of untreated group [(KD: t=15.36, P<0.001; t=7.25, P<0.001; t=14.29, P<0.001), (Ctrl: t=7.87, P<0.001; t=5.71, P<0.001; t=8.74, P<0.001)], as the binding ability of RBP-J with Foxp3 promoter increased slightly without statistically significant difference (KD: t=1.11, P>0.05; Ctrl: t=1.37, P>0.05). Simultaneously, H4Ac level of Foxp3 promoter and its binding abilities with NICD1 and p300 in KD group were still lower than those of the control group after stimulation ( t=3.86, P<0.001; t=3.42, P=0.001; t=2.85, P=0.006). ④ After incubation of PBMC from heathy children with KD serum, the proportion of Treg cells, protein level of Foxp3 and expressions of Notch1 and RBP-J in CD4 + T cells in the group treated with IVIG increased significantly compared with the untreated group ( t=7.10, P<0.001; t=10.16, P<0.001; t=8.06, P<0.001; t=9.77, P<0.001), as well as H4Ac level of Foxp3 promoter and its binding abilities with NICD1 in the group treat with IVIG were also higher than the latter ( t=7.24, P<0.001; t=8.24, P<0.001). Conclusion:Insufficiency and impaired function of Treg caused by aberrant Notch1 signaling may be the important factor contributing to immune dysfunction and vascular damage in KD.
8.Changes and significance of monocytic myeloid-derived suppressor cells during acute phase of Kawasaki disease
Lingying YU ; Guobing WANG ; Pengqiang WEN ; Jiehua MEI ; Zhongxiang QI ; Mingguo XU ; Cong LIU ; Chengrong LI
Chinese Journal of Microbiology and Immunology 2021;41(10):764-770
Objective:To investigate the changes of monocytic myeloid-derived suppressor cells (M-MDSC) in children with acute Kawasaki disease (KD) and its roles in the immunological pathogenesis of KD.Methods:A total of 38 children with acute KD were enrolled in the present study and 32 age-matched healthy children were selected as control group. The proportions of HLA-DR -CD11b + CD33 + CD15 -CD14 + M-MDSC and CD4 + CD25 + CD127 - regulatory T cells (Treg) in peripheral blood, concentrations of reactive oxygen species (ROS) and expression of arginase-1 (Arg-1), CD39, CD73, CD40, CD40L and CCR5 at protein levels were detected by flow cytometry. Quantitative real-time PCR was used to evaluate the transcription levels of inducible nitric oxide synthase (iNOS) in M-MDSC and the transcription levels of cytotoxic T-lymphocyte associated antigen 4 (CTLA4) and lymphocyte-activation gene 3 (LAG3) in Treg. Concentrations of NO, CCL3, CCL4, CCL5, IL-10 and TGF-β in the supernatants of cell culture were measured by ELISA. Results:(1) The proportion of HLA-DR -CD11b + CD33 + CD15 -CD14 + M-MDSC, the concentration of intracellular ROS and the expression of iNOS, CD39 and CD73 in M-MDSC decreased significantly in patients with acute KD as compared with those in the control group ( P<0.05), and the concentrations of NO, IL-10 and TGF-β in culture supernatant of M-MDSC were lower than those in the control group upon lipopolysaccharide (LPS) stimulation for 48 h ( P<0.05). All of the aforementioned indexes restored to some extent after intravenous immunoglobulin (IVIG) therapy ( P<0.05). No statistical differences were found in Arg-1 expression between healthy controls and patients with KD before or after IVIG therapy ( P<0.05). (2) CD40 expression on M-MDSC was significantly lower in the acute KD group than in the control group ( P<0.05). The concentrations of CCL3, CCL4 and CCL5 in the culture supernatants of M-MDSC were lower in the acute KD group than in the control group after LPS stimulation ( P<0.05). With IVIG treatment, all of the indexes were up-regulated significantly ( P<0.05), although CD40 expression was still lower in the acute KD group than in the control group ( P<0.05). (3) The proportion of CD4 + CD25 + CD127 -Treg and the expression of CTLA4, LAG3, CD40L and CCR5 reduced significantly in patients with acute KD as compared those in healthy controls ( P<0.05), and all increased remarkably after IVIG therapy ( P<0.05). Pearson correlation analysis showed a positive correlation between the proportions of M-MDSC and Treg in patients with acute KD ( r=0.58, P<0.05). Conclusions:Insufficiency and impaired function of M-MDSC might be a major cause of immune dysfunction in patients with acute KD.
9.Newborn screening and genetic analysis of methylmalonic acidemia in Shaanxi province
Ruixue ZHANG ; Chengrong SONG ; Xiaoping MA ; Yan ZHANG ; Rui WANG ; Wenwen YU ; Lihui YANG ; Xuanxing SHI ; Na CAI ; Rong QIANG
Chinese Journal of Perinatal Medicine 2021;24(2):89-96
Objective:To analyze the incidence, biochemical and molecular characteristics, and gene mutation spectrum of neonatal methylmalonic acidemia (MMA) in Shaanxi province.Methods:This study involved 146 152 newborns undergoing neonatal screening for methylmalonic acidemia by tandem mass spectrometry in Northwest Women's and Children's Hospital from January 2014 and December 2019. Clinical manifestations and follow-up data of newborns diagnosed with MMA and their acylcarnitine profiles and gene mutations were analyzed. According to whether they had elevated homocysteine or not, these patients were divided into two groups, the complicated group and the isolated MMA group. The control neonates were those excluded from having methylmalonic acid by re-examination. Kruskal-Wallis and Mann-Whitney U test was conducted for statistical analysis. Results:(1) Twenty-one cases of MMA were confirmed with an incidence of 1/6 960, including 11 cases (52.4%) of isolated MMA (isolated MMA group) and 10 (47.6%) complicated by elevated homocysteine (complicated group). Eight patients in the isolated group had symptoms within one month after birth, mainly feeding difficulties, vomiting, drowsiness, poor response and infection, and five died. Patients in the complicated group were all diagnosed before developing typical clinical symptoms, and no developmental abnormalities were reported during follow-up. (2) Blood propionyl carnitine and its ratios to acetylcarnitine and free carnitine in the isolated MMA and complicated groups were higher than those in the control group [ M (min-max), 9.26 (3.70-37.78) μmol/L and 7.27 μmol/L (3.58-13.62 μmol/L) vs 4.51 μmol/L (1.48-8.69 μmol/L), H=23.239; 1.12 (0.32-2.43) and 0.74 (0.36-1.90) vs 0.25 (0.09-0.45), H=47.061; 0.94 (0.12-1.92) and 0.56 (0.18-1.03) vs 0.17 (0.06-0.38), H=36.868; all P<0.001]. The blood methionine level in the complicated group was significantly lower than that in the isolated MMA group [7.64 μmol/L (3.40-19.25 μmol/L) vs 24.22 μmol/L (10.73-56.55 μmol/L), U=3.000, P<0.001]. (3) All 21 patients carried complex heterozygous mutations or homozygous mutations in pathogenic genes, including 15 distinct MMUT mutations and 13 distinct MMACHC mutations. In the isolated MMA group, the most common mutation was c.323G>A (p.Arg108His) in the MMUT gene with a positive rate of 13.6%, and an unreported mutation, c.1676+11A>G, with unidentified clinical significance, was also found. The most common mutations in the complicated group were c.609G>A (p.Trp203Ter) and c.567dupT (p.Ile190fs) in the MMACHC gene, and the positive rates were both 20.0%. Moreover, two unreported variants, c.430-2A>C and c.648_650delAGA (p.216_217delSEinsS), were detected and suspected to be pathogenic. Conclusions:MMA is not uncommon in Shaanxi province. Children with isolated MMA tend to be more severe clinically. The identification of hotspot mutations, including c.609G>A (p.Trp203Ter) and c.567dupT (p.Ile190fs) in MMACHC gene and c.323G>A (p.Arg108His) in MMUT gene, provides a foundation for further genetic screening, counseling, and prenatal diagnosis, and is conducive to reduce the mortality and disability rate of neonatal MMA.
10.Investigation of preoperative anemia and perioperative blood transfusion in patients with duodenal papillarycarcinoma in a tertiary hospital
Yingwei SONG ; Lijuan SONG ; Junting LIU ; Chengrong BIAN ; Hongfei ZHANG ; Ting ZHANG ; Deqing WANG
Chinese Journal of Blood Transfusion 2021;34(9):1006-1009
【Objective】 To investigate the preoperative anemia and perioperative blood transfusion in patients with duodenal papillary carcinoma who underwent Whipple surgery. 【Methods】 The clinical data of 1 959 cases with duodenal papillary carcinoma, subjected to Whipple surgery, were retrospectively analyzed. 【Results】 The rate of anemia in preoperative patients with duodenal papillary carcinoma was 54.87%(1 075/1 959). The incidence rate of anemia in the three age groups from low to high was 44.92% (≤50 years old, 190/423), 52.82% (51~64 years old, 506/958), and 65.57% (≥65 years old, 379/578) (P<0.05), and the highest rate of anemia occurred in patients aged above 65. There was a significant statistical difference among patients with different body mass index (BMI)(P<0.05). Patients with moderate or severe anemia received more red blood cells than patients with mild anemia during the perioperative period (P<0.05). The average hospitalization time of the blood transfusion patients was 27.25 days, and that of non-transfusion patients was 22.22 days (P<0.05). The amount of blood loss and hospitalization time of patients underwent laparoscopic and robotic surgery were significantly lower than those underwent open surgery patients (P <0.05). There were only 24.09%(186/772) treated with drugs for anemia intervention and the majority of patients (75.91%, 586/772) were treated with blood transfusions to interfere with anemia during hospitalization. 【Conclusion】 There are significant differences in the incidence rate of preoperative anemia among patients with duodenal papillary carcinoma who undergone Whipple surgery. Low BMI, abnormal WBC, and perioperative blood transfusion are high-risk factors for prolonged hospital stay, whereas anemia is not associated with prolonged hospital stay.

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