Synthetic biology is a crucial tool for the development of the bio-industry and bio-economy, representing a significant aspect of new quality productive forces. As a core course for graduate students in bioengineering, Synthetic Biology plays a vital role in ensuring the supply of essential talents for the development of the bio-industry in the new era. To better serve regional economic development and provide high-level talents for China's progress in the bio-industry, we analyzed typical issues encountered in the past teaching activities, set up a multi-disciplinary teaching team, optimized the course contents, adjusted the teaching mode, and mobilized students' learning interest. With the application of scientific research project as the starting point, we guided students to think and discuss deeply through the simulation of application writing and project defense, which improved students' critical thinking and innovative thinking. With industrialization as a focus, we explored a new training model combining production, education, and research through the joint practice base of the university and enterprises introduced typical cases of biomanufacturing to encourage students to engage in scientific research. The teaching reform significantly enhances the comprehensive abilities and national sentiments of graduate students. This paper hopes to serve as a reference for colleagues engaged in teaching in this field.
Synthetic Biology/education* ; Teaching ; China ; Humans

Objective To investigate the correlation between the expression of lectin galactoside binding soluble-3 binding protein(LGALS3BP),GTP enzyme activating protein SH3 functional region binding protein 1(G3BP1)and Wnt/β-catenin pathway genes in endometrial cancer(EC)tissues and its clinical prognostic significance.Methods 138 patients with EC treated in Cangzhou Central Hospital from February 2016 to February 2019 were selected.qRT-PCR was used to detect the expression of LGALS3BP mRNA,G3BP1 mRNA and Wnt/β-catenin pathway genes Wnt5a mRNA,β-catenin mRNA and matrix metalloproteinase-9(MMP-9)mRNA in cancer and adjacent tissues.The expression of LGALS3BP protein and G3BP1 protein in cancer and adjacent tissues was detected by immunohistochemistry.Pearson correlation analysis was used to analyze the correlation between LGALS3BP mRNA,G3BP1 mRNA and Wnt/β-catenin pathway genes.Kaplan-Meier curve was used to analyze the effect of LGALS3BP mRNA and G3BP1 mRNA on the survival and prognosis of EC patients.COX regression model was used to analyze the factors affecting the prognosis of EC patients.Results The expression of LGALS3BP mRNA(3.01±0.34),G3BP1 mRNA(2.87±0.33),Wnt5a mRNA(2.29±0.26),β-catenin mRNA(3.25±0.41)and MMP-9 mRNA(2.68±0.36)in EC cancer tissues were higher than those in adjacent tissues(1.10±0.23,1.06±0.24,0.84±0.17,0.88±0.26,0.69±0.17),and the differences were statistically significant(t=52.109～58.719,all P<0.001).The expression of LGALS3BP mRNA and G3BP1 mRNA in EC was positively correlated with the expression of Wnt5a mRNA,β-catenin mRNA and MMP-9 mRNA(r=0.675～0.781,all P<0.001).The expression of LGALS3BP protein(3.54±0.47 vs 0.51±0.16)and G3BP1 protein(2.84±0.44 vs 0.42±0.13)in EC cancer tissues were higher than that in adjacent tissues,and the differences were statistically significant(t=71.692,61.962,all P<0.001).The expression of LGALS3BP mRNA and G3BP1 mRNA in cancer tissues of EC patients with FIGO stage Ⅲ and lymph node metastasis were higher than that of FIGO stage Ⅰ～Ⅱ and no lymph node metastasis,and the differences were statistically significant(t=40.279～557.671,all P<0.001).The 5-year survival rate of LGALS3BP mRNA high expression group was 58.82%(40/68),which was lower than that of low expression group 94.29%(66/70),and the difference was statistically significant(Log-rank χ2=24.970,P<0.001).The 5-year survival rate of G3BP1 mRNA high expression group was 62.12%(41/66),which was lower than that of low expression group 90.28%(65/72),and the difference was statistically significant(Log-rank χ2=15.960,P<0.001).FIGO stage Ⅲ,lymph node metastasis,high expression of LGALS3BP mRNA and high expression of G3BP1 mRNA were risk factors for poor prognosis of EC patients(Wald χ2=7.847～12.054,all P<0.001).Conclusion The expression of LGALS3BP and G3BP1 mRNA is elevated in EC,both of which are associated with Wnt/β-catenin pathway genes,promoting the malignant progression of EC tumors,and are new tumor markers for evaluating the prognosis of EC patients.

Objective:To analyze the pathogenicity and clinical phenotype associated with a newly identified heterozygous variant in the ZEB1 gene that causes posterior pleomorphic corneal dystrophy (PPCD). Methods:A pedigree study was conducted.Clinical data of four people in 2 generations from one family with PPCD who visited Henan Eye Hospital in October 2023 were collected, including 3 patients. Relevant ophthalmic examinations were performed.Best corrected visual acuity, slit lamp microscopy, intraocular pressure, Pentacam corneal topography, Corvis ST corneal biomechanics analyzer, corneal endothelial microscopy, swept-source anterior segment coherence optical tomography (CASIA), laser scanning confocal microscopy, and ultra-wide-field fundus photography were performed to examine clinical phenotypes.Peripheral venous blood samples were collected from family members to extract genomic DNA, and whole exome sequencing was performed.Sanger sequencing and pedigree co-segregation analysis were carried out.Conservation analysis was performed using GERP+ + and Clustal Omega software, and the pathogenicity of the variant was assessed according to American College of Medical Genetics and Genomics (ACMG) guidelines.This study protocol adhered to the Declaration of Helsinki and was approved by the Ethics Committee of Henan Eye Hospital (No.HNEECKY-2019[15]).All subjects or guardian signed informed consent.Results:This family conformed to autosomal dominant inheritance.Under a slit-lamp microscope, corneal endothelial vesicular lesions in both eyes could be seen in the proband, her father and her brother.Under a laser scanning confocal microscope, endothelial cells were missing at the lesions, and some were crater-like changes, and some lesions were circular or elliptical vesicular, and no other systemic abnormalities were observed.The ocular and physical examination of the proband's mother showed no abnormalities.Genetic testing results showed that the proband, her father and her brother all carried the ZEB1c.790G>A (p.Gly264Arg) heterozygous variant, but her mother did not carry the variantion.Sanger sequencing verified that this variantion was co-segregated within the family.The variantion is a newly discovered missense mutation that had not been reported in the Thousand Genomes Project, Genome Aggregation Database, and ExAC database.The prediction results of the variant by MutationTaster, SIFT, PROVEAN, VESST3, DANN, FATHMM-MKL, CADD, fitCons and other software were harmful, and GERP+ +, Weblogo, Clustal Omega analysis showed that the amino acids affected by the variant were highly conservative.According to the ACMG Guidelines, this variation was possible pathogenic.Conclusions:The identification of the missense mutation c. 790G>A (p.Gly264Arg) in the ZEB1 gene within this PPCD family provides new insights into the genetic basis of PPCD and the variant may be the pathogenic variant of in this family.

Objective·To investigate the causal relationship between plasma zinc-alpha-2-glycoprotein 1(AZGP1)and heart failure(HF)by using Mendelian randomization(MR)analysis and experimental validation.Methods·A two-sample MR analysis was performed to assess the causal relationship between AZGP1 and HF by integrating large-scale genome-wide association study(GWAS)data on plasma proteins and HF.The inverse-variance weighted(IVW)method was employed as the primary analytical approach,supplemented by MR-Egger regression,weighted median,and simple median methods.Horizontal pleiotropy was tested by using MR-PRESSO global test and MR-Egger intercept analysis.Colocalization analysis was conducted to validate genetic locus overlap.Additionally,a clinical cohort(84 HF patients and 68 healthy controls)was analyzed,with plasma AZGP1 levels quantified by enzyme-linked immunosorbent assay(ELISA).Results·MR analysis showed that elevated plasma AZGP1 levels were significantly associated with reduced HF risk(OR=0.82,95%CI 0.75?0.90,P=1.70×10-5).Colocalization analysis confirmed that AZGP1 expression and HF shared causal genetic variants(posterior probability for H4=0.69).Sensitivity and reverse MR analyses supported the robustness of the results.ELISA confirmed that plasma AZGP1 levels were significantly lower in HF patients compared to healthy controls,reinforcing its protective role in HF.Conclusion·This study demonstrates AZGP1 exerts a protective causal effect on HF and may serve as a potential biomarker for HF treatment.

Objective:To analyze the pathogenicity and clinical phenotype associated with a newly identified heterozygous variant in the ZEB1 gene that causes posterior pleomorphic corneal dystrophy (PPCD). Methods:A pedigree study was conducted.Clinical data of four people in 2 generations from one family with PPCD who visited Henan Eye Hospital in October 2023 were collected, including 3 patients. Relevant ophthalmic examinations were performed.Best corrected visual acuity, slit lamp microscopy, intraocular pressure, Pentacam corneal topography, Corvis ST corneal biomechanics analyzer, corneal endothelial microscopy, swept-source anterior segment coherence optical tomography (CASIA), laser scanning confocal microscopy, and ultra-wide-field fundus photography were performed to examine clinical phenotypes.Peripheral venous blood samples were collected from family members to extract genomic DNA, and whole exome sequencing was performed.Sanger sequencing and pedigree co-segregation analysis were carried out.Conservation analysis was performed using GERP+ + and Clustal Omega software, and the pathogenicity of the variant was assessed according to American College of Medical Genetics and Genomics (ACMG) guidelines.This study protocol adhered to the Declaration of Helsinki and was approved by the Ethics Committee of Henan Eye Hospital (No.HNEECKY-2019[15]).All subjects or guardian signed informed consent.Results:This family conformed to autosomal dominant inheritance.Under a slit-lamp microscope, corneal endothelial vesicular lesions in both eyes could be seen in the proband, her father and her brother.Under a laser scanning confocal microscope, endothelial cells were missing at the lesions, and some were crater-like changes, and some lesions were circular or elliptical vesicular, and no other systemic abnormalities were observed.The ocular and physical examination of the proband's mother showed no abnormalities.Genetic testing results showed that the proband, her father and her brother all carried the ZEB1c.790G>A (p.Gly264Arg) heterozygous variant, but her mother did not carry the variantion.Sanger sequencing verified that this variantion was co-segregated within the family.The variantion is a newly discovered missense mutation that had not been reported in the Thousand Genomes Project, Genome Aggregation Database, and ExAC database.The prediction results of the variant by MutationTaster, SIFT, PROVEAN, VESST3, DANN, FATHMM-MKL, CADD, fitCons and other software were harmful, and GERP+ +, Weblogo, Clustal Omega analysis showed that the amino acids affected by the variant were highly conservative.According to the ACMG Guidelines, this variation was possible pathogenic.Conclusions:The identification of the missense mutation c. 790G>A (p.Gly264Arg) in the ZEB1 gene within this PPCD family provides new insights into the genetic basis of PPCD and the variant may be the pathogenic variant of in this family.

Objective To investigate the correlation between the expression of lectin galactoside binding soluble-3 binding protein(LGALS3BP),GTP enzyme activating protein SH3 functional region binding protein 1(G3BP1)and Wnt/β-catenin pathway genes in endometrial cancer(EC)tissues and its clinical prognostic significance.Methods 138 patients with EC treated in Cangzhou Central Hospital from February 2016 to February 2019 were selected.qRT-PCR was used to detect the expression of LGALS3BP mRNA,G3BP1 mRNA and Wnt/β-catenin pathway genes Wnt5a mRNA,β-catenin mRNA and matrix metalloproteinase-9(MMP-9)mRNA in cancer and adjacent tissues.The expression of LGALS3BP protein and G3BP1 protein in cancer and adjacent tissues was detected by immunohistochemistry.Pearson correlation analysis was used to analyze the correlation between LGALS3BP mRNA,G3BP1 mRNA and Wnt/β-catenin pathway genes.Kaplan-Meier curve was used to analyze the effect of LGALS3BP mRNA and G3BP1 mRNA on the survival and prognosis of EC patients.COX regression model was used to analyze the factors affecting the prognosis of EC patients.Results The expression of LGALS3BP mRNA(3.01±0.34),G3BP1 mRNA(2.87±0.33),Wnt5a mRNA(2.29±0.26),β-catenin mRNA(3.25±0.41)and MMP-9 mRNA(2.68±0.36)in EC cancer tissues were higher than those in adjacent tissues(1.10±0.23,1.06±0.24,0.84±0.17,0.88±0.26,0.69±0.17),and the differences were statistically significant(t=52.109～58.719,all P<0.001).The expression of LGALS3BP mRNA and G3BP1 mRNA in EC was positively correlated with the expression of Wnt5a mRNA,β-catenin mRNA and MMP-9 mRNA(r=0.675～0.781,all P<0.001).The expression of LGALS3BP protein(3.54±0.47 vs 0.51±0.16)and G3BP1 protein(2.84±0.44 vs 0.42±0.13)in EC cancer tissues were higher than that in adjacent tissues,and the differences were statistically significant(t=71.692,61.962,all P<0.001).The expression of LGALS3BP mRNA and G3BP1 mRNA in cancer tissues of EC patients with FIGO stage Ⅲ and lymph node metastasis were higher than that of FIGO stage Ⅰ～Ⅱ and no lymph node metastasis,and the differences were statistically significant(t=40.279～557.671,all P<0.001).The 5-year survival rate of LGALS3BP mRNA high expression group was 58.82%(40/68),which was lower than that of low expression group 94.29%(66/70),and the difference was statistically significant(Log-rank χ2=24.970,P<0.001).The 5-year survival rate of G3BP1 mRNA high expression group was 62.12%(41/66),which was lower than that of low expression group 90.28%(65/72),and the difference was statistically significant(Log-rank χ2=15.960,P<0.001).FIGO stage Ⅲ,lymph node metastasis,high expression of LGALS3BP mRNA and high expression of G3BP1 mRNA were risk factors for poor prognosis of EC patients(Wald χ2=7.847～12.054,all P<0.001).Conclusion The expression of LGALS3BP and G3BP1 mRNA is elevated in EC,both of which are associated with Wnt/β-catenin pathway genes,promoting the malignant progression of EC tumors,and are new tumor markers for evaluating the prognosis of EC patients.

Objective·To investigate the causal relationship between plasma zinc-alpha-2-glycoprotein 1(AZGP1)and heart failure(HF)by using Mendelian randomization(MR)analysis and experimental validation.Methods·A two-sample MR analysis was performed to assess the causal relationship between AZGP1 and HF by integrating large-scale genome-wide association study(GWAS)data on plasma proteins and HF.The inverse-variance weighted(IVW)method was employed as the primary analytical approach,supplemented by MR-Egger regression,weighted median,and simple median methods.Horizontal pleiotropy was tested by using MR-PRESSO global test and MR-Egger intercept analysis.Colocalization analysis was conducted to validate genetic locus overlap.Additionally,a clinical cohort(84 HF patients and 68 healthy controls)was analyzed,with plasma AZGP1 levels quantified by enzyme-linked immunosorbent assay(ELISA).Results·MR analysis showed that elevated plasma AZGP1 levels were significantly associated with reduced HF risk(OR=0.82,95%CI 0.75?0.90,P=1.70×10-5).Colocalization analysis confirmed that AZGP1 expression and HF shared causal genetic variants(posterior probability for H4=0.69).Sensitivity and reverse MR analyses supported the robustness of the results.ELISA confirmed that plasma AZGP1 levels were significantly lower in HF patients compared to healthy controls,reinforcing its protective role in HF.Conclusion·This study demonstrates AZGP1 exerts a protective causal effect on HF and may serve as a potential biomarker for HF treatment.

Objective:To screen the differentially expressed immune-related genes(DEIRGs)in in-stent restenosis(ISR),and to analyze the immune cell infiltration in ISR,and to clarify the mechanism of occurrence and development of ISR.Methods:The mRNA gene expression data of GSE46560 dataset samples were downloaded from the Gene Expression Omnibus(GEO),and divided into ISR group and non-ISR group.The"Limma"package in R software was used to identify the differentially expressed genes(DEGs)which were then intersected with immune-related genes(IRGs)to identify the DEIRGs in ISR;R software was used for Gene Ontology(GO)functional enrichment andalysis and Kyoto Encyclopedia of Genes and Genomes(KEGG)signaling pathway enrichment analysis on DEIRGs;the STRING database was used to construct the protein-protein interaction(PPI)network,which was visualized and analyzed for Hub genes by Cytoscape software;the receiver operating characteristic(ROC)curve of the Hub genes were plotted,and the area under the curve(AUC)was calculated and the diagnostic value was evaluated;CIBERSORT software was used to analyze the immune cell infiltration in ISR;Pearson correlation analysis was used to analyze the relationships between the immune cells and the relationships between the immune cells and key genes.Results:A total of 331 DEGs were identified(P<0.05,|log2FC|>1),including 176 upregulated genes and 155 downregulated genes,and 38 DEIRGs were obstained.The GO functional enrichment analysis results showed that the DEIRGs were mainly enriched in biological processes(BP)such as defense response,immune response,and immune system;in cellular components(CC),the DEIRGs were located primarily in the extracellular region and cytoplasmic membrane;and in molecular functions(MF),the DEIRGs were mainly involved in regulating signaling receptor binding and cytokine receptor activity.The KEGG signaling pathway enrichment analysis results indicated that the DEIRGs in ISR were primarily enriched in the phosphatidylinositol 3-kinase/protein kinase B(PI3K-AKT)and transforming growth factor-β(TGF-β)signaling pathways.In the PPI network,CD19 had the highest node among the top 10 Hub genes.Compared with non-ISR group,the expression level of the CD19 gene in the samples in ISR group was increased(P<0.05).The AUC value in the ROC curve of CD19 gene expression was 0.92(P<0.05).The immune cell infiltration analysis results showed that compared with non-ISR group,the infiltration level of T lymphocyte follicular helper(Tfh)cells in the patients in ISR group were increased(P<0.05),the infiltration levels of immature B lymphocytes,CD8+T lymphocytes,naive CD4+T lymphocytes,and M0 macrophages were increased,but the differences were not statistically significant(P>0.05),while the infiltration levels of memory B lymphocytes,activated memory CD4+T lymphocytes,regulatory T cells,resting natural killer(NK)cells,activated NK cells,monocytes,resting mast cells,and neutrophils were decreased,but the differences were not statistically significant(P>0.05).There were positive correlations between Tfh cells and M0 macrophages and resting mast cells(r=0.88,P<0.05;r=0.68,P<0.05),and there were negative correlations between Tfh cells and monocytes and neutrophils(r=-0.49,P<0.05;r=-0.42,P<0.05).Conclusion:CD19 may influence the occurrence and development of ISR by regulating the activation of the PI3K-AKT signaling pathway to affect the Tfh and B lymphocytes.CD19 can serve as a biomarker for the diagnosis of ISR.

Objective:This study aims to explore the relationship between the abundance of Lachnoclostridium genus in the gut microbiome and inflammatory markers with cognitive impairment in patients with first-episode schizophrenia. Methods:A total of 87 medication-naive patients with first-episode schizophrenia (patient group) and 87 matched healthy controls (control group) who visited the Department of Psychiatry, the First Affiliated Hospital of Zhengzhou University between January 2020 and December 2022 were selected for this study. A 24-week follow-up was conducted for the patients, and all patients received treatment with risperidone. Venous blood and fecal samples were collected from the subjects at baseline and week 24 to measure the levels of superoxide dismutase-1, erythrocyte sedimentation rate, and the abundance of Lachnoclostridium. The severity of symptoms in patients with schizophrenia was assessed using the Positive and Negative Syndrome Scale, and the cognitive function of all subjects was evaluated using MATRICS Consensus Cognitive Battery tests. The differences in the abundance of Lachnoclostridium, inflammatory markers, and cognitive scores between groups were analyzed using t-tests, and the correlations between Lachnoclostridium abundance, inflammatory markers, and cognitive scores were analyzed using Pearson correlation analysis. Results:(1) At baseline, compared with the control group, patients with first-episode schizophrenia had lower levels of superoxide dismutase-1( t=6.83, P<0.001) and total cognitive function test scores( t=6.35, P<0.001), and higher abundance of Lachnoclostridium( Z=-4.64, P<0.001). (2) At baseline, the levels of superoxide dismutase-1 in patients with first-episode schizophrenia were positively correlated with social cognition( r=0.30, P=0.005), while erythrocyte sedimentation rate was negatively correlated with information processing speed and social cognition( r=-0.23, -0.31, both P<0.050). The abundance of Lachnoclostridium genus was negatively correlated with speed of processing( r=-0.28, P=0.009), working memory( r=-0.22, P=0.040), and visual memory( r=-0.32, P=0.003). (3) After 24 weeks of risperidone treatment, the levels of superoxide dismutase-1( t=-2.07, P=0.045) and total cognitive function test scores( t=-3.47, P=0.001) increased in patients, while erythrocyte sedimentation rate( t=2.21, P=0.033) decreased. The abundance of Lachnoclostridium genus showed a decreasing trend( Z=1.52, P=0.128) and did not differ significantly from the control group( Z=1.68, P=0.094). (4) Among the 39 patients who completed the 24-week follow-up, the baseline abundance of Lachnoclostridium genus was negatively correlated with attention and vigilance( r=-0.39, P=0.014) and total cognitive function test scores( r=-0.34, P=0.032) at week 24. The baseline erythrocyte sedimentation rate was positively correlated with the differences in speed of processing, working memory, social cognition, and total cognitive function test scores between baseline and week 24( r=0.42, 0.32, 0.41, 0.36, all P<0.050). (5) At baseline, the abundance of Lachnoclostridium genus had predictive value for erythrocyte sedimentation rate( r=0.45, P=0.004), attention and vigilance( r=-0.39, P=0.014), and total cognitive function test scores( r=-0.34, P=0.032) at week 24. Conclusion:In first-episode schizophrenia patients, there is a significant correlation between the abundance of gut Lachnoclostridium and inflammation and cognitive function.

Objective:To explore the role of interaction between stearidonic acid (SDA) and fatty acid desaturase 2 ( FADS2) rs174570 genotyping in the cognitive function of schizophrenia (SCH). Methods:This study is a case-control study, patients with first-episode, drug-na?ve schizophrenia were recruited from the First Affiliated Hospital of Zhengzhou University′s Department of Psychiatry from October 2017 to October 2019. Healthy controls were recruited through advertisements and medical examinations during the same period. Peripheral blood SDA levels of the SCH patient group and the control group were measured and compared using liquid chromatograph-mass spectrometer (LC-MS), and paired sample t-test was conducted to analyze the changes in the patient group before and after treatment with risperidone. Genome-wide association study (GWAS) was used for analyzing the key enzyme of SDA, and analysis of variance was performed to evaluate the relationship between FADS2 single nucleotide polymorphism (SNP) genotyping and the level of SDA. The Positive and Negative Syndrome Scale (PANSS) and the MATRICS Consensus Cognitive Battery (MCCB) were used to assess the severity of psychotic symptoms and cognitive function, the comparison between the two groups was conducted by independent sample t-test, and the changes before and after risperidone treatment were analyzed by paired sample t-test. Linear regression analysis was performed to investigate the relationship between the interaction of SDA and FADS2 rs174570 genotyping, and cognitive impairment in SCH. Results:SDA levels were significantly lower in the SCH group compared to the control group ( t=-10.67, P<0.001). Cognitive score in patients with SCH were lower than that of HCs ( t=-10.30—-3.30, P<0.05 for all). Low levels of SDA in patients with SCH were positively correlated with the score of speed of processing (SOP; r=0.406, P<0.001) at baseline. After six months of treatment with risperidone, serum levels of SDA increased from (3.6±1.9) μmol/L to (4.4±2.3) μmol/L, and paired t-tests showed significant difference ( t=-2.29, P=0.024). The change of SDA levels before and after risperidone treatment was positively correlated with the change of SOP scores ( r=0.327, P=0.002). FADS2 rs174570 genotyping were significantly associated with SDA levels ( F=3.74, P=0.027) and cognitive function scores of SOP ( F=4.28, P=0.017), and attention/vigilance (AV; F=6.74, P=0.002). Pairwise comparisons showed that CC carriers of rs174570 genotype had higher SDA levels than CT and TT carriers ( P=0.024, and 0.048, respectively), and higher total scores of SOP, AV and MCCB than CT carriers ( P=0.006, 0.001, and 0.002, respectively). The interaction of SDA and FADS2 rs174570 genotyping were associated with cognitive function SOP scores in patients with SCH (β=1.82, P=0.029). Conclusion:The interaction of SDA and FADS2 rs174570 genotyping is associated with the cognitive function in patients with SCH.