The aim of this study is to establish an gene editing method of Mesomycoplasma hyo-pneumoniae(Mhp)based on the homologous recombination principle.The restriction enzyme di-gestion and ligation method combined with gene synthesis were used to construct a shuttle plasmid to achieve replication in both Mhp and Escherichia coli(E.coli).The pGEM?-T vector was used as the skeleton.The oriC sequence of Mhp which can achieve the replication of the plasmid in Mhp was inserted into the vector.Sequences of the Spiroplasma promoter and puromycin resistance gene were then inserted into the above constructed plasmid to screen recombinant clones.The up-stream and downstream homologous arms of p97 were constructed to initiate homologous recombination.The recA gene of E.coli is inserted to improve the efficiency of homologous recom-bination.The obtained shuttle plasmid was then delivered into Mhp by electro-transformation or chemical transformation.A shuttle plasmid,pGEM?-Mhp-oriC-p 97,which can replicate in both Mhp and E.coli was constructed.With the transformation of this plasmid,the carried puromycin gene and recA gene can be expressed,the p97 gene can be edited.Finally,the genetically unstable p97 gene mutant was initially obtained.In this study,a tool for Mhp gene editing based on the principle of homologous recombination was established,which laid a foundation for the develop-ment of tools for studying the pathogenesis of Mhp.

OBJECTIVE: Therapeutic angiogenesis has become a promising approach for treating ischemic heart disease (IHD). The present study aims to investigate the effects of Qishen Granule (QSG) on angiogenesis in myocardial ischemia (MI) and the potential mechanism. METHODS: In vivo study was conducted on rat model of myocardial infarction. QSG was performed daily at a dose of 2.352 g/kg for four weeks. Cardiac function was assessed by echocardiogram and pro-angiogenic effects were evaluated by Laser Doppler and CD31 expression. Oxygen-glucose deprivation (OGD) was applied in cultured human umbilical vein endothelial cells (HUVECs). Cell viability, wound healing and tube formation assay were used to test functions of HUVECs. ELISA and Western blots were used to assess protein expressions of bone morphogenetic protein 2-delta-like 4-notch homolog 1 (BMP2-Dll4-Notch1) signaling pathway. RESULTS: The results showed that QSG improved heart function, cardiac blood flow and microvessel density in myocardial ischemic rats. In vitro, QSG protected HUVECs by promoting the cell viability and tube formation. QSG upregulated bone morphogenetic protein-2 (BMP2) and downregulated delta-like 4 (Dll4) and notch homolog 1 (Notch1) expressions both in rats and HUVECs. CONCLUSION QSG protected against MI by promoting angiogenesis through BMP2-Dll4-Notch1 pathway. BMP2 might be a promising therapeutic target for IHD.

Toxoplasmosis is a zoonotic disease caused by infection with Toxoplasma gondii. Congenital toxoplasmosis is a common form of intrauterine infection and is associated with severe neurological sequelae such as cerebral palsy and cognitive impairment. This report presented the magnetic resonance imaging (MRI) features of a fetal intracranial toxoplasmosis case, including bilateral ventriculomegaly, multiple intracranial cystic lesions, and parenchymal calcifications, without concurrent retinal abnormalities or hepatosplenomegaly. Post-termination analyses confirmed the presence of T.gondii DNA in amniotic fluid and umbilical venous blood, with histopathology revealing necrosis and eosinophilic infiltration. MRI demonstrates superior soft-tissue resolution in evaluating the extent of cerebral lesions and parenchymal damage, underscoring its diagnostic value in fetal toxoplasmic encephalopathy.

OBJECTIVES: To investigate the ethnic origin of Chuanqing people, one of the largest unidentified ethnic groups in Guizhou, China, and analyze its genetic relationships with surrounding populations. METHODS: Based on a self-developed microhaplotype system, we conducted genotyping and analyzed the genetic distribution of microhaplotype loci and their forensic applicability in Chuanqing population in Guizhou Province. Using the microhaplotype data from different intercontinental populations and previously reported data from Han population living in Guizhou Province, we systematically investigated the genetic background of Chuanqing people through population genetic approaches, including genetic distance estimation, principal component analysis, and phylogenetic tree construction. RESULTS: Among the studied population, the number of haplotype per microhaplotype ranged from 6 to 25. The average expected heterozygosity (He), observed heterozygosity (Ho), power of discrimination (PD), and probability of exclusion (PE) were 0.8291, 0.8301, 0.9387, and 0.6593, respectively. The cumulative power of discrimination (CPD) and cumulative probability of exclusion (CPE) for these 33 loci were 1-2.62×10^-41 and 1-7.64×10^-17, respectively. Population genetic analyses revealed that the Chuanqing population had close genetic relationships with the East Asian populations, especially the local Guizhou Han population, Beijing Han population and the Han populations living in southern China. CONCLUSIONS The 33 microhaplotypes exhibit high levels of genetic diversity in the Guizhou Chuanqing population, highlighting their potentials for both forensic identification and parentage testing. The Han populations might have contributed a significant amount of genetic material to the Chuanqing population during the formation and development of the latter.
Humans ; China/ethnology* ; Ethnicity/genetics* ; Forensic Genetics/methods* ; Genetics, Population ; Genotype ; Haplotypes ; Phylogeny ; East Asian People/genetics*

OBJECTIVES: To investigate the expression of the long non-coding RNA maternally expressed gene 3 (LncRNA Meg3) in patients with the Wilson disease (WD) and its correlation with the severity of liver fibrosis and autophagy-related markers. METHODS: A total of 100 WD patients and 50 healthy individuals were enrolled from the First Affiliated Hospital of Anhui University of Chinese Medicine. Serum biomarkers, including platelet count, hyaluronic acid (HA), laminin (LN), type III procollagen N-terminal peptide (PIIINP), type IV collagen (C‑IV), alanine aminotransferase (ALT), and aspartate aminotransferase (AST), were measured, and the non-invasive indices APRI and FIB-4 were calculated. Peripheral blood levels of LncRNA Meg3, Beclin-1 and LC3B were detected using RT-qPCR, and liver stiffness (LSM) and shear wave velocity (SWV) were evaluated using two-dimensional shear wave elastography (2D-SWE). The liver tissues from 10 WD patients and 10 patients with hepatic hemangioma were examined using histochemical staining, transmission electron microscopy, and RT-qPCR. RESULTS: The expression level of LncRNA Meg3 was significantly lower, while the levels of AST, ALT, HA, LN, PIIINP, C‑IV, APRI, FIB-4, LSM and SWV were significantly higher in WD patients than in the healthy individuals (all P<0.01). LncRNA Meg3 was negatively correlated with LSM, SWV, APRI, FIB-4, Beclin-1 and LC3B (P<0.05). ROC analysis demonstrated that LncRNA Meg3 effectively discriminated >F4 stage fibrosis (AUC=0.902) with a sensitivity of 92.9% and a specificity of 83.7% at the optimal cut-off value, outperforming APRI (AUC=0.746) and FIB-4 (AUC=0.661). The liver tissues from WD patients exhibited characteristic histopathological changes and lowered expression of LncRNA Meg3, which was negatively correlated with Beclin-1 and LC3B expressions (P<0.05). Liver fibrosis staging (7 S4 cases and 3 S3 cases) was significantly associated with LSM and SWV levels (P<0.05). CONCLUSIONS The expression level of LncRNA Meg3 is significantly decreased in WD patients, which is negatively correlated with the severity of liver fibrosis and closely related to the level of autophagy.
Humans ; RNA, Long Noncoding/metabolism* ; Liver Cirrhosis/metabolism* ; Adult ; Female ; Male ; Hepatolenticular Degeneration/metabolism* ; Case-Control Studies ; Young Adult ; Adolescent ; Middle Aged

The aim of this study is to establish an gene editing method of Mesomycoplasma hyo-pneumoniae(Mhp)based on the homologous recombination principle.The restriction enzyme di-gestion and ligation method combined with gene synthesis were used to construct a shuttle plasmid to achieve replication in both Mhp and Escherichia coli(E.coli).The pGEM?-T vector was used as the skeleton.The oriC sequence of Mhp which can achieve the replication of the plasmid in Mhp was inserted into the vector.Sequences of the Spiroplasma promoter and puromycin resistance gene were then inserted into the above constructed plasmid to screen recombinant clones.The up-stream and downstream homologous arms of p97 were constructed to initiate homologous recombination.The recA gene of E.coli is inserted to improve the efficiency of homologous recom-bination.The obtained shuttle plasmid was then delivered into Mhp by electro-transformation or chemical transformation.A shuttle plasmid,pGEM?-Mhp-oriC-p 97,which can replicate in both Mhp and E.coli was constructed.With the transformation of this plasmid,the carried puromycin gene and recA gene can be expressed,the p97 gene can be edited.Finally,the genetically unstable p97 gene mutant was initially obtained.In this study,a tool for Mhp gene editing based on the principle of homologous recombination was established,which laid a foundation for the develop-ment of tools for studying the pathogenesis of Mhp.

Toxoplasmosis is a zoonotic disease caused by infection with Toxoplasma gondii. Congenital toxoplasmosis is a common form of intrauterine infection and is associated with severe neurological sequelae such as cerebral palsy and cognitive impairment. This report presented the magnetic resonance imaging (MRI) features of a fetal intracranial toxoplasmosis case, including bilateral ventriculomegaly, multiple intracranial cystic lesions, and parenchymal calcifications, without concurrent retinal abnormalities or hepatosplenomegaly. Post-termination analyses confirmed the presence of T.gondii DNA in amniotic fluid and umbilical venous blood, with histopathology revealing necrosis and eosinophilic infiltration. MRI demonstrates superior soft-tissue resolution in evaluating the extent of cerebral lesions and parenchymal damage, underscoring its diagnostic value in fetal toxoplasmic encephalopathy.

Objective:To analyze the mediating role of maladaptive cognition between family relationships and internet gaming disorder (IGD) in adolescents.Methods:From September 2021 to April 2023, a total of 52 adolescents with IGD (IGD group) from the First Affiliated Hospital of Zhengzhou University and 51 normal adolescents (HC group) from community were selected as study subjects.The severity of adolescent internet addiction was assessed using the Young internet addiction scale (YIAS).Family relationships and maladaptive cognition were evaluated by the Chinese version of family environment scale (FES-CV) and the Chinese adolescents’ maladaptive cognitions scale (CAMCS).SPSS 22.0 software was used for descriptive analysis, correlation analysis and mediation analysis.Two-sample t-tests were used for quantitative data comparison, and chi-square tests were used for qualitative data comparison. Pearson correlation analysis was conducted to analyze the relationship of variables, and the intermediate variable test was performed using Bootstrap method. Results:(1) Compared with HC group, the score of YIAS in IGD group was higher, and the difference was statistically significant((63.69±10.36), (28.96±5.20), t=21.571, P<0.05). Compared with the HC group, the conflict score of FES-CV in the IGD group was higher((4.79±2.02), (3.33±1.42), t=4.228, P<0.05), and the cohesion score of FES-CV was lower ((4.46±1.64), (5.57±1.33), t=-3.768, P<0.05).Compared with the HC group, the CAMCS score in IGD group was higher, and the difference was statistically significant((28.88±10.11), (22.98±9.11), t=3.116, P<0.05). (2) There was a positive correlation between conflict of family relationships and IGD ( r=0.504, P<0.01), and a negative correlation between cohesion of family relationships and IGD( r=-0.474, P<0.01) in IGD group. (3) Path analysis and mediation effect test showed that conflict in family relationships positively predicted IGD ( β=0.331, 95% CI=0.100-0.563) and maladaptive cognition ( β=0.372, 95% CI=0.108-0.636). Maladaptive cognitive positively predicted IGD ( β=0.464, 95% CI=0.232-0.696). Maladaptive cognition played a mediating role between conflict of family relationships and IGD, and the mediating effect value was 0.173 (95% CI=0.020-0.366), accounting for 34.33%(0.173/0.504) of the total effect. Conclusion:Family relationships are related to IGD in adolescents. Maladaptive cognition may play a mediating role between family relationships and IGD in adolescents.

Objective:To analyze the mediating role of maladaptive cognition between family relationships and internet gaming disorder (IGD) in adolescents.Methods:From September 2021 to April 2023, a total of 52 adolescents with IGD (IGD group) from the First Affiliated Hospital of Zhengzhou University and 51 normal adolescents (HC group) from community were selected as study subjects.The severity of adolescent internet addiction was assessed using the Young internet addiction scale (YIAS).Family relationships and maladaptive cognition were evaluated by the Chinese version of family environment scale (FES-CV) and the Chinese adolescents’ maladaptive cognitions scale (CAMCS).SPSS 22.0 software was used for descriptive analysis, correlation analysis and mediation analysis.Two-sample t-tests were used for quantitative data comparison, and chi-square tests were used for qualitative data comparison. Pearson correlation analysis was conducted to analyze the relationship of variables, and the intermediate variable test was performed using Bootstrap method. Results:(1) Compared with HC group, the score of YIAS in IGD group was higher, and the difference was statistically significant((63.69±10.36), (28.96±5.20), t=21.571, P<0.05). Compared with the HC group, the conflict score of FES-CV in the IGD group was higher((4.79±2.02), (3.33±1.42), t=4.228, P<0.05), and the cohesion score of FES-CV was lower ((4.46±1.64), (5.57±1.33), t=-3.768, P<0.05).Compared with the HC group, the CAMCS score in IGD group was higher, and the difference was statistically significant((28.88±10.11), (22.98±9.11), t=3.116, P<0.05). (2) There was a positive correlation between conflict of family relationships and IGD ( r=0.504, P<0.01), and a negative correlation between cohesion of family relationships and IGD( r=-0.474, P<0.01) in IGD group. (3) Path analysis and mediation effect test showed that conflict in family relationships positively predicted IGD ( β=0.331, 95% CI=0.100-0.563) and maladaptive cognition ( β=0.372, 95% CI=0.108-0.636). Maladaptive cognitive positively predicted IGD ( β=0.464, 95% CI=0.232-0.696). Maladaptive cognition played a mediating role between conflict of family relationships and IGD, and the mediating effect value was 0.173 (95% CI=0.020-0.366), accounting for 34.33%(0.173/0.504) of the total effect. Conclusion:Family relationships are related to IGD in adolescents. Maladaptive cognition may play a mediating role between family relationships and IGD in adolescents.

Objective To analyze the risk factors of malnutrition in patients with gastric cancer undergoing chemotherapy and construct a nomogram prediction model. Methods A total of 152 patients who received complete chemotherapy were selected as study subjects. After complete chemotherapy, the patients were divided into well-nourished group (n=46) and undernourished group (n=106) according to the nutritional evaluation results. Logistic regression analysis was used to screen the influencing factors of malnutrition in patients with gastric cancer undergoing chemotherapy. A nomogram model was constructed to predict the risk of malnutrition in patients with gastric cancer undergoing chemotherapy. Receiver operating characteristic (ROC) curve and calibration curve were used to verify the differentiation and consistency of the nomogram model. Results Among 152 patients with gastric cancer treated with chemotherapy, 106 were malnourished, the incidence rate was 69.74%. Male (95%CI, 1.232 to 6.292, P=0.014), no parenteral nutrition support during chemotherapy (95%CI, 1.528 to 7.781, P=0.003), hypoproteinemia (95%CI, 1.193 to 76.786, P=0.033), early satiety (95%CI, 1.259 to 82.652, P=0.030) were independent risk factors for malnutrition in patients with gastric cancer undergoing chemotherapy. The area under ROC curve was 0.757 (95%CI, 0.680 to 0.833). The slope of calibration curve was close to 1, and the Hosmer-Lemeshow goodness of fit test was good (χ²=5.834, P=0.442). Conclusion Male, lack of parenteral nutrition support during chemotherapy, hypoproteinemia and early satiety are independent risk factors for malnutrition inpatients with gastric cancer undergoing chemotherapy. The prediction nomogram model built on this basis has good differentiation and consistency.