Objective:To explore the effects and potential mechanisms of chemokine-like factor-like MARVEL transmembrane domain-containing Protein 3 (CMTM3) on the proliferation and migration of glioblastoma (GBM) cells.Methods:Using CMTM3 expression data from The Cancer Genome Atlas (TCGA) and Genotype-Tissue Expression (GTEx) databases, we analyzed the differential expression of CMTM3 in GBM tissues and its impact on the prognosis of GBM patients. Immunohistochemical staining and protein content determination of CMTM3 was performed on GBM and adjacent non-cancerous tissue samples from 11 GBM patients who underwent surgical treatment at the Affiliated Hospital of Guizhou Medical University between November 3, 2022 and March 15, 2023. Additionally, the expression of CMTM3 was validated in GBM cell lines U87, U251, LN229, and the human astrocyte (NHA) cell line using real-time quantitative polymerase chain reaction (RT-qPCR) and Western blot analyses. Stable cell lines with silenced and overexpressed CMTM3 (sh-CMTM3 group and OE-CMTM3 group) were constructed using U251 cells. The effect of CMTM3 expression on cell proliferation was assessed using the Cell Counting Kit-8 (CCK-8) assay. Flow cytometry was employed to examine the impact of CMTM3 expression on the cell cycle. Transwell assays were conducted to evaluate the influence of CMTM3 expression on cell migration. Bioinformatics analysis, Western blotting, NF-κB activation-nuclear translocation assays, and the NF-κB pathway inhibitor pyrrolidine dithiocarbamate ammonium (PDTC) were used to validate the effect of CMTM3 on the NF-κB pathway. Finally, a subcutaneous tumorigenesis assay in nude mice was performed to observe the impact of CMTM3 expression on the in vivo growth of U251 cells. Results:Bioinformatics analysis revealed that CMTM3 is highly expressed in GBM tissues. Patients with a high CMTM3 expression had lower overall survival (OS) and disease-free survival (DFS) rates compared with those with a low CMTM3 expression (with P values of 0.010 and 0.032, respectively). Among the 11 GBM pathological specimens, 10 samples exhibited higher CMTM3 protein expression levels in the cancerous tissue compared with the adjacent non-cancerous tissue. The average CMTM3 protein expression in these samples was 0.44±0.09, significantly higher than that in the adjacent non-cancerous tissues (0.12±0.02, P＜0.001). In one sample, the difference in CMTM3 protein expression between the cancerous and adjacent non-cancerous tissues was not statistically significant ( P=0.750).The RT-qPCR results showed that the mRNA expression level of CMTM3 in NHA cells was 1.0±0.1, whereas in GBM cells U87, LN229, and U251, the levels were 2.1±0.3, 3.4±0.5, and 3.7±0.8, respectively, all significantly higher than that in NHA cells (all P＜0.01). Western blot results demonstrated that the protein expression levels of CMTM3 in GBM cells U87, LN229, and U251 were 1.5±0.2, 1.8±0.2, and 1.9±0.1, respectively, also higher than that in NHA cells (0.7±0.2, all P＜0.01), with the highest level observed in U251 cells. The CCK-8 assay, Flow cytometry, and Transwell migration experiments indicated that cell viability was inhibited in the sh-CMTM3 group, with an increase in the proportion of cells in the G 0/G 1 phase ( P＜0.01) and a decrease in the S phase ( P＜0.01), and the number of migrated cells was 233.6±35.5, lower than that in the sh-NC group ( P＜0.001). Conversely, the OE-CMTM3 group showed enhanced cell viability, a reduction in the proportion of cells in the G 0/G 1 phase ( P＜0.01), and an increase in the S phase ( P＜0.01), and the number of migrated cells was 1212.0±20.8, higher than that in the OE-NC group ( P＜0.001). However, in the OE-CMTM3+PDTC group, cell viability, cell cycle distribution (G 1, S, and G 2 phases), and cell migration numbers showed no significant changes (all P＞0.05). Western blot analysis and NF-κB activation-nuclear translocation assay results indicated that in the sh-CMTM3 group, the p-p65/p65 ratio was 0.51±0.04 and the p-IκBα/IκBα ratio was 0.39±0.03, both lower than those in the sh-NC group (both P＜0.01). The cytoplasmic staining rate was (49.29±1.98)%, higher than that in the sh-NC group ( P＜0.01). In the OE-CMTM3 group, the p-p65/p65 ratio was 2.27±0.10 and the p-IκBα/IκBα ratio was 2.14±0.15, both higher than those in the OE-NC group (both P＜0.01). The cytoplasmic staining rate was (18.96±1.44)%, lower than that in the OE-NC group ( P＜0.01). In the OE-CMTM3+PDTC group, there were no significant differences in the p-p65/p65 ratio, p-IκBα/IκBα ratio, and cytoplasmic staining rate compared with the OE-NC group (all P＞0.05). The subcutaneous tumorigenesis assay in nude mice showed that the tumor volume in the sh-CMTM3 group was (408.9±96.2) mm3, smaller than that in the sh-NC group ( P=0.003). The tumor volume in the OE-CMTM3 group was (1 514.5±251.5) mm3, larger than that in the OE-NC group ( P=0.005). Conclusions:In GBM, CMTM3 is highly expressed and negatively correlated with both OS and DFS of patients. CMTM3 regulates the proliferation and migration abilities of U251 cells through the NF-κB signaling pathway.

Objective:To investigate the use of apolipoprotein B (ApoB) diagnostic algorithm for hyperlipidemia typing combined with whole exome sequencing in the precise diagnosis of familial hyperlipidemia.Methods:A retrospective observational study was conducted by collecting clinical information on all patients who attended our hospital, and had their lipid levels tested from January 2023 to May 2024, including 440 patients with low-density lipoprotein cholesterol (LDL-C)>4.10 mmol/L. Family history, current lipid levels, medication use, and comorbidities were collected by telephone follow-ups. Among them, 10 patients had a family history of hyperlipidemia. Peripheral venous blood samples were collected from patients with a family history of hyperlipidemia, and whole exome sequencing was performed.Results:According to the Fredrickson typing of WHO, 10 patients (P1 to P10) could be categorized into two groups, of which only type Ⅱa could be excluded in 6 cases, and the typing could not be determined in 4 cases. The ApoB diagnostic algorithm of hyperlipidemia typing could classify patients P1 and P2 as type Ⅱa, patients P3 to P7, P9 and P10 as type Ⅱb, and patient P8 as type Ⅴ, respectively. Whole exome sequencing detected mutations in LDLR, PCSK9, C5AR2, KIF12, ALMS1, ABCG5, COL4A3, and MTTP genes.Conclusion:The ApoB diagnostic algorithm for hyperlipidemia can be used for accurate typing of hyperlipidemia, and ApoB could be recommended as a routine lipid testing parameter. The ApoB diagnostic algorithm for hyperlipidemia combined with whole exome sequencing could be used for the accurate typing of patients with familial hyperlipidemia and defining the underlying gene mutations.

Objective:To investigate the influence of different sensory learning style preferences of officers and soldiers on satisfaction with military continuing education online courses, understand their diversified and personalized learning needs, and provide a reference for online course construction.Methods:VARK (Visual, Aural, Read/Write, Kinesthetic) questionnaire and a self-designed satisfaction questionnaire were distributed to naval officers and soldiers who participated in online courses of military continuing education. Of the 10 000 questionnaires distributed, 8 124 (81.24%) valid responses were collected. SPSS 22.0 was used for statistical analysis. The satisfaction was expressed by ( x± s). The satisfaction of participants with different sensory learning style types, tendency multiplicity, intensity, and distribution was compared using analysis of variance. Pairwise comparisons for significant differences were conducted using the Student-Newman-Keuls test at a significance level of 0.05. Results:①Significant differences were observed in the satisfaction of officers and soldiers with different types of sensory learning styles on the 14 aspects of military continuing education online courses ( F=3.41, 3.33, 1.82, 2.22, 1.84, 2.67, 2.37, 2.27, 2.68, 2.20, 2.61, 3.37, 2.86, and 2.38; all P<0.05), and the satisfaction with each aspect was in the same column. ②Significant differences were observed in the satisfaction of officers and soldiers with different tendency multiplicity in the 14 aspects of military continuing education online courses ( F=12.21, 11.92, 6.15, 6.14, 5.85, 8.29, 7.47, 8.62, 9.10, 6.51, 8.81, 12.18, 9.27, and 9.00; all P<0.05), and the officers and soldiers with quadruple styles showed relatively high satisfaction. ③Significant differences were observed in the satisfaction of officers and soldiers with different intensities of sensory learning styles in the 12 aspects of military continuing education online courses ( F=4.18, 5.27, 3.06, 2.76, 3.54, 4.23, 4.68, 2.95, 4.85, 7.66, 5.75, and 2.69; all P<0.05), except for the 2 aspects of job promotion courses as well as the richness, scientificity, novelty, and refinement of course content ( F=1.68 and 1.79; P=0.169 and 0.147). ④Significant differences were observed in the satisfaction of officers and soldiers with different distribution of sensory learning styles in the 12 aspects of military continuing education online courses ( F=4.33, 5.16, 4.57, 2.84, 3.51, 3.70, 4.24, 2.71, 4.29, 6.07, 5.19, and 2.47; all P<0.05), except for the 2 aspects of job promotion courses as well as the richness, scientificity, novelty, and refinement of course content ( F=1.61 and 2.35, P=0.168 and 0.052). Conclusions:The satisfaction with military continuing education online courses was influenced by the sensory preferences of learners, and the satisfaction of officers and soldiers with multiple styles was relatively high. The overall satisfaction of officers and soldiers was generally high, but special attention should be paid to groups with low satisfaction. The advantage of personalized teaching in online education should be utilized and multiple measures should be taken to improve the quality of online education.

Objective:To investigate the use of apolipoprotein B (ApoB) diagnostic algorithm for hyperlipidemia typing combined with whole exome sequencing in the precise diagnosis of familial hyperlipidemia.Methods:A retrospective observational study was conducted by collecting clinical information on all patients who attended our hospital, and had their lipid levels tested from January 2023 to May 2024, including 440 patients with low-density lipoprotein cholesterol (LDL-C)>4.10 mmol/L. Family history, current lipid levels, medication use, and comorbidities were collected by telephone follow-ups. Among them, 10 patients had a family history of hyperlipidemia. Peripheral venous blood samples were collected from patients with a family history of hyperlipidemia, and whole exome sequencing was performed.Results:According to the Fredrickson typing of WHO, 10 patients (P1 to P10) could be categorized into two groups, of which only type Ⅱa could be excluded in 6 cases, and the typing could not be determined in 4 cases. The ApoB diagnostic algorithm of hyperlipidemia typing could classify patients P1 and P2 as type Ⅱa, patients P3 to P7, P9 and P10 as type Ⅱb, and patient P8 as type Ⅴ, respectively. Whole exome sequencing detected mutations in LDLR, PCSK9, C5AR2, KIF12, ALMS1, ABCG5, COL4A3, and MTTP genes.Conclusion:The ApoB diagnostic algorithm for hyperlipidemia can be used for accurate typing of hyperlipidemia, and ApoB could be recommended as a routine lipid testing parameter. The ApoB diagnostic algorithm for hyperlipidemia combined with whole exome sequencing could be used for the accurate typing of patients with familial hyperlipidemia and defining the underlying gene mutations.

Objective To identify the mutation in pathogenic genes by analyzing the clinical features and genotype of a family with Al-port syndrome,in order to provide a theoretical basis in genetic counseling for guidance on the future pregnancies in this couple.Meth-ods Based on closely combined thorough re-examination for the report of previous whole-exome sequencing of the proband and the mu-tation site information,Sanger sequencing verification was conducted in the proband's family members.Additionally,a comprehensive assessment of the proband's clinical manifestations and family history was performed,and the previous whole-exome sequencing report was reanalyzed accordingly.Results In terms of the heterozygous mutation(NM_0000924:c.3289+1G＞A)in intron 35 of COL4A4 gene carried by the proband,this mutation was identified in the proband's father(Ⅲ7),grandmother(Ⅱ6),aunt(Ⅲ6),and two great-uncle(Ⅱ2 and Ⅱ5).All of them exhibited clinical manifestations carried the COL4A4:c.3289+1G＞A heterozygous mutation.However,no mutation was detected in the proband's mother(Ⅲ8),great-grandmother(Ⅰ2),great-aunt(Ⅱ3),and great-uncle(Ⅱ4),grandfather(Ⅱ7)who were clinically unaffected.Additionally,a heterozygous mutation(COL4A3:NM_000914:c.1956A＞G)was identified in ex-on 27 of the COL4A3 gene in the proband.Her mother(Ⅲ 8),the other grandmother(Ⅱ9)and aunt(Ⅲ9)all carried the mutation of COL4A3 but had no clinical manifestation.However,her father(Ⅲ7)did not carry this mutation.Conclusion The splicing site muta-tion COL4A4:NM_0000924:c.3289+1G＞A should be confirmed as the pathogenic cause of Alport syndrome in this family.The combi-nation of whole-exome high-throughput sequencing and Sanger sequencing can effectively diagnose Alport syndrome and provide genetic counseling for the couple's next pregnancy.

Objective:To investigate the influence of different sensory learning style preferences of officers and soldiers on satisfaction with military continuing education online courses, understand their diversified and personalized learning needs, and provide a reference for online course construction.Methods:VARK (Visual, Aural, Read/Write, Kinesthetic) questionnaire and a self-designed satisfaction questionnaire were distributed to naval officers and soldiers who participated in online courses of military continuing education. Of the 10 000 questionnaires distributed, 8 124 (81.24%) valid responses were collected. SPSS 22.0 was used for statistical analysis. The satisfaction was expressed by ( x± s). The satisfaction of participants with different sensory learning style types, tendency multiplicity, intensity, and distribution was compared using analysis of variance. Pairwise comparisons for significant differences were conducted using the Student-Newman-Keuls test at a significance level of 0.05. Results:①Significant differences were observed in the satisfaction of officers and soldiers with different types of sensory learning styles on the 14 aspects of military continuing education online courses ( F=3.41, 3.33, 1.82, 2.22, 1.84, 2.67, 2.37, 2.27, 2.68, 2.20, 2.61, 3.37, 2.86, and 2.38; all P<0.05), and the satisfaction with each aspect was in the same column. ②Significant differences were observed in the satisfaction of officers and soldiers with different tendency multiplicity in the 14 aspects of military continuing education online courses ( F=12.21, 11.92, 6.15, 6.14, 5.85, 8.29, 7.47, 8.62, 9.10, 6.51, 8.81, 12.18, 9.27, and 9.00; all P<0.05), and the officers and soldiers with quadruple styles showed relatively high satisfaction. ③Significant differences were observed in the satisfaction of officers and soldiers with different intensities of sensory learning styles in the 12 aspects of military continuing education online courses ( F=4.18, 5.27, 3.06, 2.76, 3.54, 4.23, 4.68, 2.95, 4.85, 7.66, 5.75, and 2.69; all P<0.05), except for the 2 aspects of job promotion courses as well as the richness, scientificity, novelty, and refinement of course content ( F=1.68 and 1.79; P=0.169 and 0.147). ④Significant differences were observed in the satisfaction of officers and soldiers with different distribution of sensory learning styles in the 12 aspects of military continuing education online courses ( F=4.33, 5.16, 4.57, 2.84, 3.51, 3.70, 4.24, 2.71, 4.29, 6.07, 5.19, and 2.47; all P<0.05), except for the 2 aspects of job promotion courses as well as the richness, scientificity, novelty, and refinement of course content ( F=1.61 and 2.35, P=0.168 and 0.052). Conclusions:The satisfaction with military continuing education online courses was influenced by the sensory preferences of learners, and the satisfaction of officers and soldiers with multiple styles was relatively high. The overall satisfaction of officers and soldiers was generally high, but special attention should be paid to groups with low satisfaction. The advantage of personalized teaching in online education should be utilized and multiple measures should be taken to improve the quality of online education.

Objective To identify the mutation in pathogenic genes by analyzing the clinical features and genotype of a family with Al-port syndrome,in order to provide a theoretical basis in genetic counseling for guidance on the future pregnancies in this couple.Meth-ods Based on closely combined thorough re-examination for the report of previous whole-exome sequencing of the proband and the mu-tation site information,Sanger sequencing verification was conducted in the proband's family members.Additionally,a comprehensive assessment of the proband's clinical manifestations and family history was performed,and the previous whole-exome sequencing report was reanalyzed accordingly.Results In terms of the heterozygous mutation(NM_0000924:c.3289+1G＞A)in intron 35 of COL4A4 gene carried by the proband,this mutation was identified in the proband's father(Ⅲ7),grandmother(Ⅱ6),aunt(Ⅲ6),and two great-uncle(Ⅱ2 and Ⅱ5).All of them exhibited clinical manifestations carried the COL4A4:c.3289+1G＞A heterozygous mutation.However,no mutation was detected in the proband's mother(Ⅲ8),great-grandmother(Ⅰ2),great-aunt(Ⅱ3),and great-uncle(Ⅱ4),grandfather(Ⅱ7)who were clinically unaffected.Additionally,a heterozygous mutation(COL4A3:NM_000914:c.1956A＞G)was identified in ex-on 27 of the COL4A3 gene in the proband.Her mother(Ⅲ 8),the other grandmother(Ⅱ9)and aunt(Ⅲ9)all carried the mutation of COL4A3 but had no clinical manifestation.However,her father(Ⅲ7)did not carry this mutation.Conclusion The splicing site muta-tion COL4A4:NM_0000924:c.3289+1G＞A should be confirmed as the pathogenic cause of Alport syndrome in this family.The combi-nation of whole-exome high-throughput sequencing and Sanger sequencing can effectively diagnose Alport syndrome and provide genetic counseling for the couple's next pregnancy.

Objective:To explore the effects and potential mechanisms of chemokine-like factor-like MARVEL transmembrane domain-containing Protein 3 (CMTM3) on the proliferation and migration of glioblastoma (GBM) cells.Methods:Using CMTM3 expression data from The Cancer Genome Atlas (TCGA) and Genotype-Tissue Expression (GTEx) databases, we analyzed the differential expression of CMTM3 in GBM tissues and its impact on the prognosis of GBM patients. Immunohistochemical staining and protein content determination of CMTM3 was performed on GBM and adjacent non-cancerous tissue samples from 11 GBM patients who underwent surgical treatment at the Affiliated Hospital of Guizhou Medical University between November 3, 2022 and March 15, 2023. Additionally, the expression of CMTM3 was validated in GBM cell lines U87, U251, LN229, and the human astrocyte (NHA) cell line using real-time quantitative polymerase chain reaction (RT-qPCR) and Western blot analyses. Stable cell lines with silenced and overexpressed CMTM3 (sh-CMTM3 group and OE-CMTM3 group) were constructed using U251 cells. The effect of CMTM3 expression on cell proliferation was assessed using the Cell Counting Kit-8 (CCK-8) assay. Flow cytometry was employed to examine the impact of CMTM3 expression on the cell cycle. Transwell assays were conducted to evaluate the influence of CMTM3 expression on cell migration. Bioinformatics analysis, Western blotting, NF-κB activation-nuclear translocation assays, and the NF-κB pathway inhibitor pyrrolidine dithiocarbamate ammonium (PDTC) were used to validate the effect of CMTM3 on the NF-κB pathway. Finally, a subcutaneous tumorigenesis assay in nude mice was performed to observe the impact of CMTM3 expression on the in vivo growth of U251 cells. Results:Bioinformatics analysis revealed that CMTM3 is highly expressed in GBM tissues. Patients with a high CMTM3 expression had lower overall survival (OS) and disease-free survival (DFS) rates compared with those with a low CMTM3 expression (with P values of 0.010 and 0.032, respectively). Among the 11 GBM pathological specimens, 10 samples exhibited higher CMTM3 protein expression levels in the cancerous tissue compared with the adjacent non-cancerous tissue. The average CMTM3 protein expression in these samples was 0.44±0.09, significantly higher than that in the adjacent non-cancerous tissues (0.12±0.02, P＜0.001). In one sample, the difference in CMTM3 protein expression between the cancerous and adjacent non-cancerous tissues was not statistically significant ( P=0.750).The RT-qPCR results showed that the mRNA expression level of CMTM3 in NHA cells was 1.0±0.1, whereas in GBM cells U87, LN229, and U251, the levels were 2.1±0.3, 3.4±0.5, and 3.7±0.8, respectively, all significantly higher than that in NHA cells (all P＜0.01). Western blot results demonstrated that the protein expression levels of CMTM3 in GBM cells U87, LN229, and U251 were 1.5±0.2, 1.8±0.2, and 1.9±0.1, respectively, also higher than that in NHA cells (0.7±0.2, all P＜0.01), with the highest level observed in U251 cells. The CCK-8 assay, Flow cytometry, and Transwell migration experiments indicated that cell viability was inhibited in the sh-CMTM3 group, with an increase in the proportion of cells in the G 0/G 1 phase ( P＜0.01) and a decrease in the S phase ( P＜0.01), and the number of migrated cells was 233.6±35.5, lower than that in the sh-NC group ( P＜0.001). Conversely, the OE-CMTM3 group showed enhanced cell viability, a reduction in the proportion of cells in the G 0/G 1 phase ( P＜0.01), and an increase in the S phase ( P＜0.01), and the number of migrated cells was 1212.0±20.8, higher than that in the OE-NC group ( P＜0.001). However, in the OE-CMTM3+PDTC group, cell viability, cell cycle distribution (G 1, S, and G 2 phases), and cell migration numbers showed no significant changes (all P＞0.05). Western blot analysis and NF-κB activation-nuclear translocation assay results indicated that in the sh-CMTM3 group, the p-p65/p65 ratio was 0.51±0.04 and the p-IκBα/IκBα ratio was 0.39±0.03, both lower than those in the sh-NC group (both P＜0.01). The cytoplasmic staining rate was (49.29±1.98)%, higher than that in the sh-NC group ( P＜0.01). In the OE-CMTM3 group, the p-p65/p65 ratio was 2.27±0.10 and the p-IκBα/IκBα ratio was 2.14±0.15, both higher than those in the OE-NC group (both P＜0.01). The cytoplasmic staining rate was (18.96±1.44)%, lower than that in the OE-NC group ( P＜0.01). In the OE-CMTM3+PDTC group, there were no significant differences in the p-p65/p65 ratio, p-IκBα/IκBα ratio, and cytoplasmic staining rate compared with the OE-NC group (all P＞0.05). The subcutaneous tumorigenesis assay in nude mice showed that the tumor volume in the sh-CMTM3 group was (408.9±96.2) mm3, smaller than that in the sh-NC group ( P=0.003). The tumor volume in the OE-CMTM3 group was (1 514.5±251.5) mm3, larger than that in the OE-NC group ( P=0.005). Conclusions:In GBM, CMTM3 is highly expressed and negatively correlated with both OS and DFS of patients. CMTM3 regulates the proliferation and migration abilities of U251 cells through the NF-κB signaling pathway.

Objective:To investigate the expression of hippocampal synapse-related proteins including synaptophysin (SYN), postsynaptic density protein 95 (PSD95) and growth-associated protein 43 (GAP43) in rats with epilepsy accompanied by depression.Methods:The 3-month-old female clean grade SD rats were selected for the experiment.Lithium chloride pilocarpine was used to establish an epileptic rat model. Rats with successful epilepsy models were divided into epileptic depressive group (EWD group)and epileptic group with 10 in each group based on whether they were accompanied by depression. Furthermore, ten rats with matched body mass were taken as the depressive group and 10 were taken as control group. As for the depressive group rats, chronic unpredictable mild stress stimulation combined with orphanage was adopted to establish a model of depression.The depressive behaviors of rats were evaluated by body mass, sucrose preference test and open field test. Immunohistochemical staining and Western blot were used to detect the expression of SYN, PSD95 and GAP43 proteins in rat hippocampal tissue. SPSS 17.0 software was used for data statistical analysis, repeated measurement ANOVA was used for behavioral results, one-way ANOVA was used for inter group comparison of protein expression data, and LSD test was used for further pairwise comparison.Results:As for the body mass, there was significant interaction effect between the time and group among the 4 groups ( F=7.33, P<0.01). On the 8th day and the 29th day, the body weight of rats in the EWD group and the depressive group were lower than those in the epilepsy group (all P<0.05). The body weight of rats in the EWD group on the 29th day was lower than that on the first day ( P<0.05). As for the sucrose preference rates, there was significant interaction effect between the time and group among the 4 groups( F=2.67, P<0.05). The sucrose preference rate of EWD group on the15th and 29th day were lower than that on the first day (both P<0.05). The results of the open field test showed that the interaction effects of the number of vertical standing times( F=2.74) and the number of horizontal movement lattices ( F=1.76) both were not significant (both P>0.05), but both the time effect and group effect were significant (vertical standing times: Ftime=4.35, P<0.05, Fgroup=25.64, P<0.01; horizontal movement lattices: Ftime=12.75, P<0.01, Fgroup=21.37, P<0.01). The immunohistochemical results showed that there was a statistically significant difference in the number of positive cells expressing synaptic proteins SYN, PSD95 and GAP43 among the four groups of rats ( F=93.85, 58.66, 98.84, all P<0.05). The numbers of positive cells of SYN (11.73±4.30), PSD95 (24.47±7.58) and GAP43 (9.40±3.50) in the epilepsy group were lower than those in the control group ((51.00±15.39), (55.60±13.17) and (29.53±4.05)) (all P<0.05). The numbers of positive cells of SYN (5.80±3.53), PSD95 (12.87±4.03) and GAP43 (5.33±3.50) in the EWD group were lower than those in the depressive group ((11.33±3.22), (48.13±12.69) and (15.47±5.21) )(all P<0.05). Western blot results showed that there were statistically significant differences in the expression of synaptic proteins SYN, PSD95 and GAP43 among the four groups of rats ( F=13.19, 9.38, 16.80, all P<0.05). The expression levels of SYN, PSD95 and GAP43 in the epilepsy group were lower than those in the control group (all P<0.05). The expression levels of SYN, PSD95 and GAP43 in the EWD group were lower than those in the epilepsy group and the depressive group (all P<0.05). Conclusion:The low expression of SYN, PSD95 and GAP43 proteins in the hippocampus of rats with epilepsy accompanied by depression may be related to their pathogenesis.

Objective:To explore the genetic etiology of a child with primary hypertrophic osteoarthropathy(PHO).Methods:A child who was admitted to Zhongnan Hospital of Wuhan University on July 27, 2021 was selected as the study subject. Genomic DNA was extracted from peripheral blood samples of the child and his parents and subjected to whole exome sequencing. Suspected splicing variant was verified by Sanger sequencing of family members. In vitro function was validated through a minigene assay, whilst the suspected exonic deletion was validated by long-fragment PCR. This study was approved by the Children′s Hospital Affiliated to Zhengzhou University (Ethics No. 2023-K-011). Results:Whole exome sequencing revealed that the child has harbored compound heterozygous variants of HPGD gene, including a heterozygous deletion (Exon 3 del) derived from his father and a splicing variant (c.421+ 1G>T) derived from his mother. Long-fragment PCR verified that the child and his father had both harbored a 7 565 bp heterozygous deletion (c.218-1304_324+ 6156del), whilst the minigene assay proved that the splicing variant has resulted in skipping of exon 4. Conclusion:The heterozygous c. 218-1304_324+ 6156del deletion and the c. 421+ 1G>T splicing variant of the HPGD gene probably underlay the pathogenesis in this child. Above finding has enriched the mutational spectrum of the HPGD gene and provided a basis for genetic counseling and prenatal diagnosis for this family.