1.Effects of Weicanqing Formula (微残清方) on Malic Enzyme 2-Mediated Bone Marrow Immunemetabolic Homeostasis in Acute Myeloid Leukemia Model Mice
Chenyang FAN ; Lixiang YAN ; Xiaogang HAO ; Xinli ZHOU ; Reaila JIANATI ; Yifei GUO ; Gengda ZHU ; Zhexin SHI
Journal of Traditional Chinese Medicine 2026;67(12):1315-1322
ObjectiveTo
2.Genetic analysis of a Chinese pedigree with rare mosaic 11q partial duplication and a literature review.
Lili ZHOU ; Chenyang XU ; Hao WU ; Sheng HUANG ; Xueqin XU ; Xiaohua TANG
Chinese Journal of Medical Genetics 2025;42(1):94-101
OBJECTIVE:
To explore the genetic characteristics of a Chinese pedigree with rare mosaic 11q partial duplication and its pathogenetic mechanisms.
METHODS:
A pedigree which underwent prenatal diagnosis at Wenzhou Central Hospital between September 25, 2015 and November 30, 2023 was selected for the study. Clinical data were collected from the pedigree. Peripheral blood samples from the parents, amniotic fluid from the fetus, and peripheral blood sample from the neonate were obtained. Genetic testing was carried out by using G-banded chromosomal karyotyping and single nucleotide polymorphism array (SNP-array) technology. Relevant literature was searched in the CNKI, Wanfang Data Knowledge Service Platform, and PubMed databases to summarize the clinical phenotypes of patients with 11q partial duplication. This study was approved by the Medical Ethics Committee of Wenzhou Central Hospital (Ethics No. L2024-07-080).
RESULTS:
The pregnant woman (G3) had a history of adverse pregnancy outcomes. During her first pregnancy (G1), prenatal ultrasound indicated intrauterine growth restriction and a Dandy-Walker variant. Follow-up at 8 years of age showed developmental delays and mild intellectual disability. During her second pregnancy (G2), prenatal ultrasound revealed nasal bone hypoplasia, and the pregnancy was terminated at 23rd gestational week. During her third pregnancy (G3), all prenatal tests were normal, and the neonate showed normal growth and development at 4 months of age. The karyotype of amniotic fluid of her first pregnancy was 46,X?, and the SNP-array analysis of neonatal peripheral blood showed arr[GRCh37/hg19]11q13.4q25(70432450_134607121)×2~3, with a mosaicism rate being approximately 40%. The karyotype for her second pregnancy was 46,X?,rec(11)dup(11q)inv(11)(p15q13)dmat[6]/46,X?[27], and the SNP-array result was arr[GRCh38]11q13.4q25(71406636_135067522)×2~3, with a mosaicism rate being approximately 75%. The karyotype for her third pregnancy was 46,X?,inv(11)(p15q13)mat, and the SNP-array result was arr(XN)×1,(1~22)×2. The karyotype of the woman was 46,XX,inv(11)(p15q13), and that of her husband was 46,XY. A review of 12 similar cases (including G1) from the literature revealed that the common clinical phenotypes of 11q partial duplication included intellectual disability (12/12), developmental delay (12/12), ear abnormalities (12/12), microcephaly (10/12), seizures (8/12), hypotonia (8/12), and congenital heart malformations (7/12).
CONCLUSION
Mosaic partial duplication of 11q may underlie the genetic etiology of this pedigree. The pregnant woman is a carrier of an inversion on chromosome 11, which might have formed the mosaic 11q partial duplication through meiotic errors and mitotic trisomy rescue mechanisms during reproduction.
Adult
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Female
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Humans
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Male
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Pregnancy
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China
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Chromosome Duplication
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Chromosomes, Human, Pair 11/genetics*
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East Asian People/genetics*
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Karyotyping
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Mosaicism
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Pedigree
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Polymorphism, Single Nucleotide
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Prenatal Diagnosis
3.Genetic analysis of a Chinese pedigree with rare mosaic 11q partial duplication and a literature review
Lili ZHOU ; Chenyang XU ; Hao WU ; Sheng HUANG ; Xueqin XU ; Xiaohua TANG
Chinese Journal of Medical Genetics 2025;42(1):94-101
Objective:To explore the genetic characteristics of a Chinese pedigree with rare mosaic 11q partial duplication and its pathogenetic mechanisms.Methods:A pedigree which underwent prenatal diagnosis at Wenzhou Central Hospital between September 25, 2015 and November 30, 2023 was selected for the study. Clinical data were collected from the pedigree. Peripheral blood samples from the parents, amniotic fluid from the fetus, and peripheral blood sample from the neonate were obtained. Genetic testing was carried out by using G-banded chromosomal karyotyping and single nucleotide polymorphism array (SNP-array) technology. Relevant literature was searched in the CNKI, Wanfang Data Knowledge Service Platform, and PubMed databases to summarize the clinical phenotypes of patients with 11q partial duplication. This study was approved by the Medical Ethics Committee of Wenzhou Central Hospital (Ethics No. L2024-07-080).Results:The pregnant woman (G 3) had a history of adverse pregnancy outcomes. During her first pregnancy (G 1), prenatal ultrasound indicated intrauterine growth restriction and a Dandy-Walker variant. Follow-up at 8 years of age showed developmental delays and mild intellectual disability. During her second pregnancy (G 2), prenatal ultrasound revealed nasal bone hypoplasia, and the pregnancy was terminated at 23rd gestational week. During her third pregnancy (G 3), all prenatal tests were normal, and the neonate showed normal growth and development at 4 months of age. The karyotype of amniotic fluid of her first pregnancy was 46, X? , and the SNP-array analysis of neonatal peripheral blood showed arr[GRCh37/hg19]11q13.4q25(70432450_134607121)×2~3, with a mosaicism rate being approximately 40%. The karyotype for her second pregnancy was 46, X? , rec(11)dup(11q)inv(11)(p15q13)dmat[6]/46, X? [27], and the SNP-array result was arr[GRCh38]11q13.4q25(71406636_135067522)×2~3, with a mosaicism rate being approximately 75%. The karyotype for her third pregnancy was 46, X? , inv(11)(p15q13)mat, and the SNP-array result was arr(XN)×1, (1~22)×2. The karyotype of the woman was 46, XX, inv(11)(p15q13), and that of her husband was 46, XY. A review of 12 similar cases (including G 1) from the literature revealed that the common clinical phenotypes of 11q partial duplication included intellectual disability (12/12), developmental delay (12/12), ear abnormalities (12/12), microcephaly (10/12), seizures (8/12), hypotonia (8/12), and congenital heart malformations (7/12). Conclusion:Mosaic partial duplication of 11q may underlie the genetic etiology of this pedigree. The pregnant woman is a carrier of an inversion on chromosome 11, which might have formed the mosaic 11q partial duplication through meiotic errors and mitotic trisomy rescue mechanisms during reproduction.
4.Genetic analysis of a Chinese pedigree with rare mosaic 11q partial duplication and a literature review
Lili ZHOU ; Chenyang XU ; Hao WU ; Sheng HUANG ; Xueqin XU ; Xiaohua TANG
Chinese Journal of Medical Genetics 2025;42(1):94-101
Objective:To explore the genetic characteristics of a Chinese pedigree with rare mosaic 11q partial duplication and its pathogenetic mechanisms.Methods:A pedigree which underwent prenatal diagnosis at Wenzhou Central Hospital between September 25, 2015 and November 30, 2023 was selected for the study. Clinical data were collected from the pedigree. Peripheral blood samples from the parents, amniotic fluid from the fetus, and peripheral blood sample from the neonate were obtained. Genetic testing was carried out by using G-banded chromosomal karyotyping and single nucleotide polymorphism array (SNP-array) technology. Relevant literature was searched in the CNKI, Wanfang Data Knowledge Service Platform, and PubMed databases to summarize the clinical phenotypes of patients with 11q partial duplication. This study was approved by the Medical Ethics Committee of Wenzhou Central Hospital (Ethics No. L2024-07-080).Results:The pregnant woman (G 3) had a history of adverse pregnancy outcomes. During her first pregnancy (G 1), prenatal ultrasound indicated intrauterine growth restriction and a Dandy-Walker variant. Follow-up at 8 years of age showed developmental delays and mild intellectual disability. During her second pregnancy (G 2), prenatal ultrasound revealed nasal bone hypoplasia, and the pregnancy was terminated at 23rd gestational week. During her third pregnancy (G 3), all prenatal tests were normal, and the neonate showed normal growth and development at 4 months of age. The karyotype of amniotic fluid of her first pregnancy was 46, X? , and the SNP-array analysis of neonatal peripheral blood showed arr[GRCh37/hg19]11q13.4q25(70432450_134607121)×2~3, with a mosaicism rate being approximately 40%. The karyotype for her second pregnancy was 46, X? , rec(11)dup(11q)inv(11)(p15q13)dmat[6]/46, X? [27], and the SNP-array result was arr[GRCh38]11q13.4q25(71406636_135067522)×2~3, with a mosaicism rate being approximately 75%. The karyotype for her third pregnancy was 46, X? , inv(11)(p15q13)mat, and the SNP-array result was arr(XN)×1, (1~22)×2. The karyotype of the woman was 46, XX, inv(11)(p15q13), and that of her husband was 46, XY. A review of 12 similar cases (including G 1) from the literature revealed that the common clinical phenotypes of 11q partial duplication included intellectual disability (12/12), developmental delay (12/12), ear abnormalities (12/12), microcephaly (10/12), seizures (8/12), hypotonia (8/12), and congenital heart malformations (7/12). Conclusion:Mosaic partial duplication of 11q may underlie the genetic etiology of this pedigree. The pregnant woman is a carrier of an inversion on chromosome 11, which might have formed the mosaic 11q partial duplication through meiotic errors and mitotic trisomy rescue mechanisms during reproduction.
5.Identification and Analysis of NnOMT and NnNMT Gene Families in Nelumbo nucifera
Zhuyin CHEN ; Yuetong YU ; Chenyang HAO ; Mingru LIU ; Sha CHEN
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(3):152-160
ObjectiveThe biosynthetic pathways of benzylisoquinoline alkaloids(BIAs) in Nelumbo nucifera are of great theoretical and economic value. In this paper, N. nucifera O-methyltransferase(NnOMT) and N. nucifera N-methyltransferase(NnNMT) gene families were identified and analyzed by bioinformatics in order to facilitate the biosynthetic pathway of BIAs in N. nucifera. MethodBased on the whole genome of N. nucifera, UniPort and National Center for Biotechnology Information(NCBI) databases were used to identify the NnOMT and NnNMT gene families of N. nucifera, and analyze their physicochemical properties and subcellular localization, then TBtools, MEME, MEGA 11.0, FigTree 1.4.4 and other tools were used to analyze the phylogeny, sequence characteristics, gene structure, functional annotation and cis-acting elements of NnOMT and NnNMT genes identified in the previous stage. ResultA total of 61 NnOMT and NnNMT genes were identified in this paper, the number of amino acids encoded by these genes ranged from 168 aa to 580 aa, the isoelectric point ranged from 4.76 to 9.16, and the relative molecular weight ranged from 18 699.52 Da to 64 934.53 Da, most of which showed acidic and mostly hydrophilic proteins. There were 10 conserved motifs, Kyoto Encyclopedia of Genes and Genomes(KEGG) analysis enriched a total of 12 pathways, including metabolism, biosynthesis of phenylpropane and isoquinoline alkaloids, etc. And Visualization of Gene Ontology(GO) enrichment results showed that 61 NnOMT and NnNMT genes were annotated to 32 items, which included 16 molecular functions[such as reduced nicotinamide adenine dinucleotide(NADH) activity and exopeptidase activity] and 16 biological processes(such as metabolic process of carbon tetrachloride, anaerobic carbon tetrachloride metabolic process and responses to exogenous biological stimuli). There were a variety of cis-acting elements in the promoter regions of NnOMT and NnNMT genes, mainly promoter and enhancer regions element, light responsive element and methyl jasmonate responsive element. ConclusionIn this study, a comprehensive bioinformatics analysis of 61 NnOMT and NnNMT genes is carried out based on the genome data of N. nucifera, which lays a foundation for research on the gene structure and function of NnOMT and NnNMT gene families, and provides a reference for biosynthetic pathway elucidation of BIAs in N. nucifera.
6.Construction of double-layer bone-on-a-chip containing bone matrix
Hao ZHANG ; Chenyang ZHOU ; Yan HU ; Xiao CHEN ; Xiaolin WANG ; Jiacan SU
Chinese Journal of Trauma 2024;40(1):20-28
Objective:To construct a double-layer bone-on-a-chip containing bone matrix, with which the process of osteoblast and osteoclast differentiation in vitro is stimulated, aiming to provide a new platform for the development of osteoporosis medications. Methods:Software WorkSoild was used to design the double-layer and double-channel bone-on-a-chip and the template was fabricated by photolithography. With polydimethylsiloxane (PDMS) as the raw material, the main body of the chip was prepared by mold fabrication. The inlets and outlets of the four channels of the culture room were separated with bovine cortex bones and sealed with liquid storage columns. In the chip verification experiment, chips were divided into osteogenic and osteoclastic induction groups and osteogenic and osteoclastic control groups. In the osteogenic and osteoclastic induction groups, precursor cells of mouse embryonic osteoblast, MC3T3-E1 and mouse macrophage RAW264.7 were inoculated on the chip separately. Osteogenic induction lasted 14 days and osteoclastic induction 7 days. MC3T3-E1 cells and RAW264.7 cells were not induced in the osteogenic and osteoclastic control groups. The following indicators were observed: (1) Appearance and sealing performance of the chip: After the chip was prepared, photos were taken to observe its appearance and sealing tests were conducted to observe its sealing performance. (2) Biocompatibility: At 3 days after MC3T3-E1 cells were inoculated onto the chip and cultured and at 1, 3 and 5 days after RAW264.7 cells were inoculated onto the chip and cultured, the cell survival was observed with calcein acetoxymethyl ester/propidium iodide (AM/PI) staining and Cell Counting Kit 8 (CCK-8). (3) Osteogenic differentiation: Alkaline phosphatase (ALP) staining and alizarin red staining were performed on the cells in the osteogenic induction group to observe the osteogenic induction. RNA was collected from the osteogenic induction group and the osteogenic control group, the expression of osteoblast marker Runt-related transcription factor 2 (RUNX2), osteocalcin (OCN) and type I collagen (COL1A1) was detected by real-time florescent quantitative PCR (qPCR), and the differentiation degree and osteogenic ability of osteoblasts were observed. (4) Osteoclast differentiation: tartrate-resistant acid phosphatase (TRAP) staining was performed on cells in the osteoclastic induction group to observe osteoclast differentiation. RNA was extracted from the osteoclastic induction group and the osteoclastic control group for qPCR of osteoclast differentiation-related genes, and the expression levels of the osteoclast marker gene TRAP, cathepsin K (CTSK) and dendritic cell specific transmembrane protein (DC-STAMP) were detected.Results:The double-layer bone-on-a-chip containing bone matrix was 3 cm×3 cm in size and transparent as a whole. The structure of the system on the chip system was compact and had no seepage. It was shown by calcein AM/PI staining that at 3 days after MC3T3-E1 cells and RAW264.7 cells were cultured, very few red fluorescent dead cells were found. CCK-8 test showed that within 5 days after being cultured, the cell viability was all above 90%, indicating that the biocompatibility of the chip was good and the cells could survive and proliferate normally. The results of ALP and alizarin red staining showed that MC3T3-E1 cells successfully differentiated into osteoblasts and produced calcified nodules in the osteogenic induction group at 14 days after the induction. The qPCR results showed that the relative expression level of RUNX2 in MC3T3-E1 cells in the osteogenic induction group was 4.98±0.74, which was significantly higher than that of the control group (0.99±0.03) ( P<0.01). The relative expression level of OCN in MC3T3-E1 cells was 7.98±0.76, which was significantly higher than that of the control group (1.00±0.06) ( P<0.01). The relative expression level of COL1A1 in MC3T3-E1 cells was 7.07±0.56, which was significantly higher than that of the control group (0.97±0.03) ( P<0.01). The TRAP staining results showed that the RAW264.7 cells in the osteoclastic induction group differentiated to giant multinucleated osteoclasts, and TRAP protein was expressed in large quantity in the osteoclasts. The results of qPCR showed that the relative expression level of TRAP in RAW264.7 cells in the osteoclastic induction group was 3.35±0.37, which was significantly higher than that of the control group (1.01±0.06) ( P<0.01). The relative expression level of CTSK in RAW264.7 cells was 3.46±0.79, which was significantly higher than that of the control group (1.01±0.05) ( P<0.01). The relative expression level of DC-STAMP in RAW264.7 cells was 1.92±0.12, which was significantly higher than that of the control group (0.98±0.08) ( P<0.01). Conclusions:The double-layer bone-on-a-chip containing bone matrix is compact in structure, can be cultured in vitro for a long time, has good biocompatibility and can be used for inducing osteogenic and osteoclast differentiation. Therefore, it is expected to provide a new research platform for exploring the mechanism of osteoporosis and medication screening.
7.Characteristics of renal oxidative stress injuries in rats with high-voltage electric burns and the intervention effects of breviscapine
Congying LI ; Xuegang ZHAO ; Jiawen HAO ; Chenyang GE ; Mengyuan LU ; Jing ZHANG ; Qingfu ZHANG ; Jianke FENG ; Lihong TU
Chinese Journal of Burns 2024;40(8):746-755
Objective:To explore the characteristics of renal oxidative stress injuries in rats with high-voltage electric burns and the intervention effects of breviscapine.Methods:This study was an experimental study. One hundred and sixty 8-10-week-old male Sprague Dawley rats were divided into sham injury group, electric burn group, saline group, low breviscapine group, middle breviscapine group, and high breviscapine group, with 60 rats in each of the sham injury group and electric burn group, 10 rats in each of the other 4 groups, respectively. The rats in sham injury group and electric burn group were divided into 10 rats at each time point, including post injury hour (PIH) 0 (immediately), 8, 24, 48, and 72, and post injury week (PIW) 1. The rats in sham injury group were not conducted with electrical current to cause sham injury. The rats in the other 5 groups were caused high-voltage electric burns. The rats in sham injury group and electric burn group were not treated after injury. The rats in saline group, low breviscapine group, middle breviscapine group, and high breviscapine group were intraperitoneally injected with 5 mL/kg normal saline or 0.4, 1.6, and 4.0 g/L breviscapine, repeated every 24 h until PIH 72. After the model was successfully made, 14 rats died, including 1, 2, 2, and 1 rat (s) at PIH 24, 48, and 72 and PIW 1 in electric burn group, 4, 1, 2, and 1 rat (s) at PIH 72 in saline group, low breviscapine group, middle breviscapine group, and high breviscapine group, respectively. The kidney tissue collected from rats in the 6 groups was weighed and the kidney/body weight ratio was calculated. The left upper pole tissue of kidney was collected from each 4 rats in sham injury group, and in electric burn group at PIH 8, 24, 48, and 72 and PIW 1, and in saline group, low breviscapine group, middle breviscapine group, and high breviscapine group at PIH 72. The renal tubular and renal interstitial injury was evaluated by a semi-quantitative histological scoring system after hematoxylin-eosin staining. The inferior vena cava blood samples were collected from rats in the 6 groups to measure the serum creatinine levels via sarcosine oxidase method, and serum urea nitrogen levels via urease method. The right renal cortices were collected from rats in the 6 groups to measure the catalase (CAT) activity in the supernatant of renal tissue via molybdic acid method, and the levels of advanced oxidation protein product (AOPP) and Klotho protein in the supernatant of renal tissue via enzyme-linked immunosorbent assay.Results:At PIH 8, 48, and 72 and PIW 1, the kidney/body weight ratios of rats in electric burn group were significantly higher than those in sham injury group (with t values of -0.52, -3.75, -4.05, and -2.25, respectively, P<0.05). At PIH 72, compared with those in electric burn group, saline group, low breviscapine group, and middle breviscapine group, the kidney/body weight ratio of rats in high breviscapine group was significantly decreased (with P values all <0.05). Compared with those in sham injury group, the renal tubular and renal interstitial injury scores of rats in electric burn group at PIH 48 and 72 and PIW 1 were significantly increased ( P<0.05). Compared with those in electric burn group at PIH 8 and 24, the renal tubular and renal interstitial injury score of rats in electric burn group at PIW 1 was significantly increased (with P values all <0.05). At PIH 72, the renal tubular and renal interstitial injury scores of rats in the 5 groups of rats with electric burns were similar ( P>0.05). At PIH 8, 24, 48, and 72 and PIW 1, the levels of serum creatinine and serum urea nitrogen of rats in electric burn group were significantly higher than those in sham injury group (with Z values of -2.00, -2.37, -2.62, -2.67, -3.67, -2.34, -3.11, -3.43, -3.11, and -3.51, respectively, P<0.05). Compared with that in electric burn group at PIH 0, the levels of serum creatinine of rats in electric burn group at PIH 72 and PIW 1 were significantly increased ( P<0.05). Compared with that in electric burn group at PIH 8, the levels of serum creatinine of rats in electric burn group at PIH 72 and PIW 1 were significantly increased ( P<0.05). Compared with that in electric burn group at PIH 24, the level of serum creatinine of rats in electric burn group at PIW 1 was significantly increased ( P<0.05). At PIH 72, the levels of serum creatinine of rats in the 5 groups of rats with electric burns were similar ( P>0.05). Compared with that in electric burn group, the levels of serum urea nitrogen of rats in low breviscapine group, middle breviscapine group, and high breviscapine group were significantly decreased ( P<0.05). Compared with that in saline group, the levels of serum urea nitrogen in middle breviscapine group and high breviscapine group were significantly decreased ( P<0.05). At PIH 48 and 72 and PIW 1, the CAT activities in the supernatant of renal tissue of rats in electric burn group were significantly lower than those in sham injury group (with Z values of -2.22, -2.13, and -3.51, respectively, P<0.05). At PIH 8, 24, 48, and 72 and PIW 1, the levels of AOPP in the supernatant of renal tissue of rats in electric burn group were significantly higher than those in sham injury group (with Z values of -2.00, -3.15, -2.71, -2.04, and -2.33, respectively, P<0.05). At PIH 0-PIW 1, the levels of Klotho protein in the supernatant of renal tissue of rats in sham injury group and electric burn group were all similar ( P>0.05). Compared with that in electric burn group at PIH 0, the CAT activities in the supernatant of renal tissue of rats in electric burn group at PIH 72 and PIW 1 and the levels of Klotho protein in the supernatant of renal tissue of rats in electric burn group at PIH 48 and 72 and PIW 1 were significantly decreased ( P<0.05). Compared with that in electric burn group at PIH 8, the CAT activities in the supernatant of renal tissue of rats in electric burn group at PIH 72 and PIW 1 and the levels of Klotho protein in the supernatant of renal tissue of rats in electric burn group at PIH 48 and 72 and PIW 1 were significantly decreased ( P<0.05). Compared with that in electric burn group at PIH 24, the CAT activities in the supernatant of renal tissue of rats in electric burn group at PIH 72 and PIW 1 were significantly decreased ( P<0.05). Compared with that in electric burn group at PIH 48, the CAT activity in the supernatant of renal tissue of rats in electric burn group at PIW 1 was significantly decreased ( P<0.05). At PIH 72, the levels of Klotho protein in the supernatant of renal tissue of rats in the 5 groups of rats with electric burns were similar ( P<0.05). Compared with 14.6 (12.6, 23.6) U/mgprot in electric burn group, the CAT activities in the supernatant of renal tissue of rats in low breviscapine group (20.5 (18.0, 39.8) U/mgprot), middle breviscapine group (24.9 (14.7, 28.9) U/mgprot), and high breviscapine group (28.0 (21.9, 39.1) U/mgprot) were significantly increased ( P<0.05). Compared with 15.7 (13.7, 25.6) U/mgprot in saline group, the CAT activities in the supernatant of renal tissue of rats in middle breviscapine group and high breviscapine group were significantly increased ( P<0.05). Compared with that in low breviscapine group, the CAT activity in the supernatant of renal tissue of rats in high breviscapine group was significantly increased ( P<0.05). Compared with those in electric burn group and saline group, the levels of AOPP in the supernatant of renal tissue of rats in middle breviscapine group and high breviscapine group were significantly decreased ( P<0.05). Conclusions:After high-voltage electric burns, oxidative stress injury occur in the kidneys of rats, which is aggravated with time extension. Breviscapine can alleviate oxidative stress injuries in the kidneys of rats with high-voltage electric burns.
8.Vector construction and protein preparation of long QT syndrome-related C-terminal lobe of calmodulin mutant E141G
Dongxue SHAO ; Chenyang ZHANG ; Miaomiao YE ; Fan CHEN ; Liying HAO
Journal of China Medical University 2024;53(11):967-971
Objective To construct a prokaryotic expression vector of of the long QT syndrome(LQTS)associated C-terminal lobe of calmodulin(CaM)mutant E141G(C-lobeE141G)and to identify the expression,purification,and activity of C-lobeE141G.Methods A cDNA fragment was inserted into a PGEX-6p-3 plasmid vector and transferred into Escherichia coli BL21 receptor cells,and glutathione-S-trans-ferase(GST)fusion protein was induced by isopropyl thio-β-D galactoside(IPTG).Glutathione-Sepharose 4B beads were used to separate and purify GST-C-lobeE141G.After removing the GST label with protease,the purity and concentration of purified C-lobeE141G were detected using SDS-PAGE and BCA,respectively.The activity of purified C-lobeE141G was detected using the GST pull-down method and patch clamp technique.Results GST-C-lobeE141G fusion protein was highly expressed,and C-lobeE141G with high purity and concentration was obtained.The purified C-lobeE141G protein not only bound to CaV1.2 calcium channels,but also rescued the channel activity from run-down in the ventricular myocytes of rat hearts.Conclusion This study successfully constructed a prokaryotic expression vector of C-lobeE141G,which provides a material basis for the study of the mechanism of LQTS mediated by C-lobe mutations in CaM.
9.The effect of triglyceride-glucose index on the severity of depression in patients with depression
Man JIN ; Hao LIANG ; Xueru ZHANG ; Shipan ZHANG ; Chenyang GAO ; Xiaona CUI ; Litao LI ; Peiyuan LYU
Chinese Journal of Behavioral Medicine and Brain Science 2024;33(12):1106-1110
Objective:To investigate the effect of triglyceride-glucose index (TyG) on the severity of depression in patients with depression.Methods:The clinical data of 653 patients with depression admitted to Hebei General Hospital from January to December 2021 were retrospectively analyzed. The patients were categorized into mild-moderate depression group ( n=324) and severe depression group ( n=329) according to the 17-item Hamilton depression scale (HAMD-17) score. Patients' demographics and data on fasting plasma glucose (FPG), triglyceride (TG), total cholesterol, high-density lipoprotein, and low-density lipoprotein were extracted from the electronic medical records, and TyG was calculated. Statistical analyses were performed by SPSS 25.0 software.The Mann-Whitney U test was used to compare the differences between the two groups. Spearman correlation analysis and binary Logistic regression analysis were used to investigate the relationship between TyG level and depression severity. Results:(1) In the severe depression group, FPG( 956.9 (118.9) mg/L vs 865.0 (1 44.2) mg/L), TG(1 275.8 (1 027.8) mg/L vs 983.5 (766.4) mg/L), total cholesterol (2 007.0 (477.6) mg/L vs 1 730.5 (547.2) mg/L), high-density lipoprotein (514.3 (168.2) mg/L vs 464.0 (138.2) mg/L), low-density lipoprotein (1 264.5 (326.8) mg/L vs 1 088.6 (412.8) mg/L), and TyG (8.72 (0.77) vs 8.37 (0.78)) were higher than those in the mild-moderate depression group, and the differences were statistically significant (all P<0.05).(2) Both TyG level and HAMA score were positively correlated with HAMD-17 score ( r=0.231, 0.586, both P<0.05); (3) The results of binary logistic regression analysis showed that TyG ( B=1.116, OR=3.054, 95% CI=2.058-4.531, P<0.05) was a risk factor for the severity of depression in patients with depression. Conclusion:The higher the TyG, the more severe the severity of depression in patients. Insulin resistance may affect the severity of depression in patients with depression.
10.The effect of triglyceride-glucose index on the severity of depression in patients with depression
Man JIN ; Hao LIANG ; Xueru ZHANG ; Shipan ZHANG ; Chenyang GAO ; Xiaona CUI ; Litao LI ; Peiyuan LYU
Chinese Journal of Behavioral Medicine and Brain Science 2024;33(12):1106-1110
Objective:To investigate the effect of triglyceride-glucose index (TyG) on the severity of depression in patients with depression.Methods:The clinical data of 653 patients with depression admitted to Hebei General Hospital from January to December 2021 were retrospectively analyzed. The patients were categorized into mild-moderate depression group ( n=324) and severe depression group ( n=329) according to the 17-item Hamilton depression scale (HAMD-17) score. Patients' demographics and data on fasting plasma glucose (FPG), triglyceride (TG), total cholesterol, high-density lipoprotein, and low-density lipoprotein were extracted from the electronic medical records, and TyG was calculated. Statistical analyses were performed by SPSS 25.0 software.The Mann-Whitney U test was used to compare the differences between the two groups. Spearman correlation analysis and binary Logistic regression analysis were used to investigate the relationship between TyG level and depression severity. Results:(1) In the severe depression group, FPG( 956.9 (118.9) mg/L vs 865.0 (1 44.2) mg/L), TG(1 275.8 (1 027.8) mg/L vs 983.5 (766.4) mg/L), total cholesterol (2 007.0 (477.6) mg/L vs 1 730.5 (547.2) mg/L), high-density lipoprotein (514.3 (168.2) mg/L vs 464.0 (138.2) mg/L), low-density lipoprotein (1 264.5 (326.8) mg/L vs 1 088.6 (412.8) mg/L), and TyG (8.72 (0.77) vs 8.37 (0.78)) were higher than those in the mild-moderate depression group, and the differences were statistically significant (all P<0.05).(2) Both TyG level and HAMA score were positively correlated with HAMD-17 score ( r=0.231, 0.586, both P<0.05); (3) The results of binary logistic regression analysis showed that TyG ( B=1.116, OR=3.054, 95% CI=2.058-4.531, P<0.05) was a risk factor for the severity of depression in patients with depression. Conclusion:The higher the TyG, the more severe the severity of depression in patients. Insulin resistance may affect the severity of depression in patients with depression.

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