1.Bioinformatic analysis of TCGA database based on INPP4B gene expression in hepatocellular carcinoma and its experimental validation
Limei WEN ; Yali GUO ; Wenmei MA ; Taotao XUE ; Ruoyu GENG ; Chong MA ; Xinhong ZHANG ; Jianhua YANG
Journal of Jilin University(Medicine Edition) 2025;51(6):1618-1629
Objective:To discuss the expression and clinical significance of inositol polyphosphate-4-phosphatase type Ⅱ(INPP4B)gene in hepatocellular carcinoma(HCC)based on The Cancer Genome Atlas(TCGA)database and experimental verification with clinical samples.Methods:Based on data from 424 clinical samples in the TCGA database(including 374 HCC tissues and 50 paracarcinoma tissues),Kaplan-Meier method and Cox regression analysis were used to evaluate the relationship between INPP4B gene and the clinical characteristics and survival prognosis of the HCC patients.The correlations between INPP4B gene and the number of 24 types of immune cells,matrix,immune cell infiltration and tumor purity in tumor tissue,and the expression level of the high-frequency mutant gene tumor protein 53(TP53)in HCC were analyzed.The clinicopathological data and paraffin-embedded tissue sections of 60 HCC patients treated with surgical resection from December 2022 to December 2023 were collected.According to clinical diagnosis,they were divided into poorly differentiated group(HCC-L group),moderately differentiated group(HCC-M group)and well-differentiated group(HCC-H group),with 20 cases in each group;20 patients during the same period who underwent biopsy and were pathologically diagnosed as non-tumor were selected as normal group,and their clinicopathologic data and liver tissue paraffin sections were collected.HE staining was used to observe the pathomorphology of HCC tissue and normal liver tissue of the subjects in various groups;immunohistochemistry method was used to detect the expressions of Ki-67 and INPP4B proteins in the HCC tissue and normal liver tissue of the subjects in various groups.Results:The TCGA database analysis results showed that compared with normal tissue,the expression level of INPP4B mRNA in HCC tissue was significantly increased(P<0.01).Compared with INPP4B low expression group,the overall survival(OS)of the patients in INPP4B high expression group was significantly prolonged(P<0.05).The univariate Cox regression analysis results showed that tumor stage,pathological stage,tumor status and residual tumor had impacts on OS of the HCC patients(P<0.05).The univariate regression analysis results showed that the INPP4B prognostic risk model score ratio was HR=0.781,95%confidence interval(CI):0.552-1.105,P=0.168.The AUC value for the impact of INPP4B on OS of the HCC patients was 0.558,indicating that the INPP4B gene prognostic risk model had certain predictive value in survival prognosis.The INPP4B mRNA expression level was not correlated with TNM stage,stage,patient gender,age,race or body mass index(BMI)(P>0.05).In tumor tissue with high and low INPP4B expression,22 types of immune cells showed statistically significant differences(P<0.05);the INPP4B mRNA expression level was positively correlated with the number of 23 types of immune cells except T helper(Th)17 cells(r>0),among which all Th cells except natural killer(NK)CD56+cells were statistically significant(P<0.01);INPP4B was significantly correlated with matrix(r=0.475),immune cell infiltration(r=0.641)and tumor purity(r=0.599)in tumor tissue(P<0.01).INPP4B was correlated with TP53(r=0.287,P<0.01).The HE staining results showed that clear and complete lobular structure,neatly arranged cells and slight inflammatory cell infiltration were observed in liver tissue of the subjects in normal group;completely destroyed lobular structure,significant hepatocellular steatosis,massive inflammatory cell infiltration,and lesions such as ballooning degeneration and small cell hyperplasia in some cells were observed in HCC tissue of the patients in HCC-L,HCC-M and HCC-H groups,and the lower the HCC differentiation degree,the more severe the tissue destruction;The immunohistochemistry results showed that compared with normal group,the expression levels of Ki-67 protein in HCC tissue of the patients in HCC-L,HCC-M and HCC-H groups were significantly increased(P<0.01),and the lower the differentiation degree of the HCC patients,the higher the Ki-67 positive rate.Brownish-yellow granules evenly distributed in the cells and INPP4B protein was highly expressed in liver tissue of the subjects in normal group;compared with normal group,the expression levels of INPP4B protein in HCC tissue of the patients in HCC-L,HCC-M and HCC-H groups were significantly decreased(P<0.01),and the lower the differentiation degree of the HCC tissue,the lower the INPP4B positive rate.Conclusion:INPP4B is a protective factor for the prognosis of HCC patients;as a new tumor suppressor gene,INPP4B may become a potential target for new drug screening in HCC treatment.
2.Polymerized gold nanorods applied to the diagnosis of Angiostrongylus cantonensis infection
Wenmei ZHANG ; Yufang KONG ; Shungao MA ; Jingwei SHEN ; Xiuhua ZUO ; Yimei YANG
International Journal of Laboratory Medicine 2025;46(15):1807-1813
Objective To construct an assay for early infection diagnosis of Angiostrongylus cantonensis based on gold nanorod polymerization.Methods Stable gold nanorods were synthesized by the gold seed growth method,and labeled with different concentrations of sulfhydrylated crude and purified antigens of lar-vae and adults of Angiostrongylus cantonensis,and their excretory and secretory antigens,and then scanned the longitudinal surface plasmon resonance(LSPR)of the stable gold nanorods by ultraviolet-visible(UV-Vis)spectrophotometry,and screened for the optimal labeled antigens for the detection of different infection time after infection of rats with Angiostrongylus cantonensis.The displacement changes were screened to se-lect the best labeled antigens for the detection of serum antibodies and positive sera of series of dilution gradi-ents at different infection times(5,7,14,21 d)after infection with Angiostrongylus cantonensis in rats,and at the same time,the enzyme-linked immunosorbent assay(ELISA)was set up for the same test.Kappa test was used to compare the consistency of the two assays.Results Gold nanorods with stable aspect ratio were suc-cessfully prepared.The gold nanorods labeled with 10 μg/mL of adult purified antigen had a maximum LSPR shift of 40 nm,and were able to detect serum antibodies in rats 5 d after mild,moderate and severe infection with Angiostrongylus cantonensis,as well as positive sera at a maximum dilution of 1∶600.The ELISA was able to detect serum antibodies in rats after 14 d of mild infection,and 7 d of moderate and severe infection,as well as positive sera at a maximum dilution of 1∶200.The ELISA detected positive serum antibodies in rats after 14 d of mild infection and 7 d of moderate and severe infection,as well as in rats at a maximum dilution of 1∶200.The Kappa value of the two methods was 0.750(P<0.01),and the results of the two methods had strong consistency.Conclusion A polymerized gold nanorod assay for early and rapid diagnosis of An-giostrongylus cantonensis infection is successfully constructed.
3.Effect of bone marrow mesenchymal stem cell on the expression of IL-10 and TNF-α in mice with ischemia-reperfusion acute kidney injury
Chenyu LIN ; Wen CHEN ; Xihui MA ; Xiangrui KONG ; Wenmei FAN ; Yong HAN ; Li XIAO ; Bingyi SHI
Organ Transplantation 2021;12(5):563-
Objective To evaluate the effect of bone marrow mesenchymal stem cell (BMSC) on the expression of interleukin (IL)-10 and tumor necrosis factor (TNF)-α in mice with ischemia-reperfusion acute kidney injury (IR-AKI). Methods All mice were randomly divided into the sham operation group (control group), ischemia-reperfusion injury group (IRI group) and BMSC treatment group (BMSC group), with 6 mice in each group, respectively. The renal function and pathological changes of mice were detected. The cell apoptosis of renal tissues of mice was determined. The expression levels of serum IL-10 and TNF-α of mice were quantitatively measured. The mouse BMSC was randomly divided into the control and hypoxia-reoxygenation groups (IRI group), and the expression levels of IL-10 and TNF-α in cell supernatant were determined. Results The renal structure of mice was normal in the control group, severe damage was observed in the IRI group, and mild damage occurred in the BMSC group. Compared with the control group, the renal tissue injury scores were significantly higher in the IRI and BMSC groups (both
4.Correlation of childhood trauma and cognitive function in patients with schizophrenia
Wenmei FANG ; Daomin ZHU ; Zicong ZHAO ; Yuting MA ; Jing ZHAO ; Anzhen WANG ; Xialong CHENG ; Boyu ZHANG ; Kai WANG ; Yi DONG
Chinese Journal of Behavioral Medicine and Brain Science 2020;29(11):995-1000
Objective:To explore the relationship between the childhood trauma and neruocognition in patients with schizophrenia.Methods:Sixty-two patients with schizophrenic were selected from Anhui mental health center, and sixty-three community health controls were selected. All subjects were assessed with the childhood trauma questionnaire (CTQ), Wisconsin card sorting Test (WCST), attention network test (ANT), verbal fluency test (VFT) and digit span test (DST). SPSS 17.0 was used for statistical analysis. t-test was used to compare the measurement data of normal distribution and Mann-Whitney U test was used to compare the measurement data of non-normal distribution. Spearman correlation analysis was used to analyze the relationship between CTQ score and cognitive function score. Results:Compared with health controls(34.00(30.00, 37.00), 6.00 (5.00, 7.00), 5.00(5.00, 5.00), 5.00(5.00, 5.00), 9.00(6.00, 11.00), 7.00(6.00, 10.00)), the total score of CTQ, subscores of emotional abuse, physical abuse, sexual abuse, emotional neglect and physical neglect in patients with schizophrenia were significantly increased (48.50(37.75, 57.00), 9.00(6.00, 12.25), 7.00(5.00, 9.25), 5.50(5.00, 7.25), 13.00 (9.00, 16.25), 11.00(8.00, 13.00)) ( Z=-4.781--6.724, all P<0.01). Compared with the control group, the number of WCST classification completed in the patient group was lower, while the number of wrong answers, continuous answers and persistent errors increased ( Z=-5.655--6.060, all P< 0.01). The correct rate of ant decreased, but the reaction time increased ( Z=-5.796, -6.094, all P< 0.01). VFT and DST scores were decreased ( Z=-3.492--8.499, both P< 0.01). In patients with schizophrenia, CTQ sexual abuse subscore were negatively correlated with completed categories scores ( r=-0.384) and positively correlated with total errors ( r=0.360), perseverative responses( r=0.394) and perseverative errors ( r=0.381) on WCST(all P<0.01). CTQ physical neglect scores were negatively correlated with the ANT correct ratio( r=-0.400) and conflict resolution( r=-0.417) (all P<0.01). CTQ emotional neglect scores were negatively correlated with VFT scores( r=-0.345) ( P<0.01). The significant associations remained after controlling for age, education and PANSS scores. Conclusion:Patients with schizophrenia experience more traumatic events in their early years and have extensive cognitive defects. The childhood trauma has negative effects on cognitive flexibility, attention, memory and speech function in patients with schizophrenia.However, the positive correlation between childhood trauma and executive conflict of attention network needs to be further verified and explored.
5.Activation of CD8+T cells regulated by γ-aminobutyric acid and its receptors
Wenmei FAN ; Yu GAO ; Yujie SUN ; Xihui MA ; Xiuyun HE ; Li XIAO ; Bingyi SHI
Organ Transplantation 2017;8(2):144-148
Objective To evaluate the effect of γ-aminobutyric acid (GABA) and its receptors upon the proliferation of CD8+T cells.Methods The splenic CD8+T cells of Balb/c mice were obtained by CD8+f cell magnetic bead sorting kit.Under the effect of CD3/CD28-activated magnetic bead,CD8+T cells were stimulated by different concentrations of GABA.5-bromo-2-deoxyuridine (BrdU) labeling and flow cytometry were performed to detect the proliferation of CD8+T cells.The expression levels of GABA-A and GABA-B receptor before and after CD8+T cell activation were compared by fluorescent quantitative real-time polymerase chain reaction (PCR).Result Flow cytometry result revealed that GABA could inhibit the proliferation of activated CD8+T cells,manifested as significant decrease in the quantity of CD152+CD8+T cells.Fluorescent quantitative real-time PCR demonstrated that GABA-A receptor subtypes α2,α6 and GABA-B receptor subtype 1a were expressed only when the CD8+T cells were activated.After CD8+T cell activation,the quantity of GABA-A receptor subtypes α3,αs,β2,β3,γ1,γ2 and θ were significantly increased,whereas the quantity of GABA-B2R and GABA-B1b did not significantly differ before and after CD8+T cell activation.Conclusions GABA can suppress the proliferation of activated CD8+T cells.The activation of CD8+T cells is regulated by GABA receptors.However,the underlying mechanism remains to be elucidated.
6.Changes of erythrocytes surface molecule CD35,CD58 and CD59 expression in recipients infected with cytomegalovirus after renal transplantation
Xiangrui KONG ; Li XIAO ; Wen CHEN ; Wenmei FAN ; Jian BAI ; Yu GAO ; Xihui MA ; Lili BI ; Bingyi SHI
Organ Transplantation 2016;7(6):473-489
Objective To investigate the change rules and its significance of erythrocytes surface molecule CD35 , CD58 and CD59 expression in recipients infected with cytomegalovirus (CMV)after renal transplantation. Methods Eighty-two recipients undergoing allogeneic renal transplantation were selected and divided into the negative (n=21 )and positive CMV groups (n=61 )based on the qualitative detection of CMV-pp65 antigen in peripheral blood. According to the results of CMV-pp65 (+)leucocyte count,all 61 patients in positive CMV group were further divided into low (n=55)and high active infection subgroups (n =6 ). Healthy adults were recruited into the normal control group (n =30 ). The expression levels of CMV-pp65 antigen,erythrocytes surface molecule CD35,CD58 and CD59 were measured by flow cytometry. Results Compared with normal control group,the expression levels of erythrocytes surface molecule CD35 , CD58 and CD59 in the positive CMV group were significantly down-regulated,and the CD35 and CD59 expression in the negative CMV group were considerably down-regulated (all P<0. 05 ). Compared with negative CMV group,the expression levels of CD58 and CD59 in the positive CMV group were significantly down-regulated (both P<0. 05 ). The expression levels of CD35 and CD59 in the high active infection subgroup were significantly lower than those in the low active infection subgroup (both P<0. 05 ). Conclusions The more severe active CMV infection after renal transplantation,the lower expression of erythrocytes surface molecule CD35,CD58 and CD59,hinting that red cell immune dysfunction is probably involved with active CMV infection.
7.Analysis of health inspection and related professional settings in colleges of China
Wenmei CAO ; Fan ZHANG ; Beilei ZHANG ; Gang CHEN ; Sha MA ; Wei LIU
Chinese Journal of Medical Education Research 2014;13(11):1081-1086
Objective To study the cultivation status of health inspection and the setting of the related major of this subject in colleges in China.Methods The setting situation of health inspection in undergraduate and junior colleges were searched from the professional information database of higher vocational schools,information query system of Chinese university and official websites of 7 junior colleges and 167 undergraduate colleges.Excel 2007 was used for inputting and sorting out data,and the data were combed and analyzed.Results The setting of college level of health inspection was approved in 2004,and there were 6 record colleges in 2014.The undergraduate level of related major was set in 2012 and only 1 college recruited students.Among 167 undergraduate universities,only 2 universities carried out professional education of health inspection,74 universities carried out professional education of health law and 65 universities carried out professional education of health management.Conclusion The subject of health inspection has been set in undergraduate and junior colleges for a short time and there are a few admissions; Professional education of preventive medicine,health law,health management and health inspection are relatively extensive.
8.γ-aminobutyric acid B receptor (GABABR)ameliorated liver fibrosis by inhibiting hepatic cell migration
Wenmei FAN ; Bingyi SHI ; Kai FENG ; Xihui MA ; Hongshan WEI ; Haiyan HUANG ; Xiuyun HE
Chinese Journal of Hepatobiliary Surgery 2012;18(8):627-630
Objective To investigate the role of r-aminobutyric acid B receptor in the development of liver fibrosis.Methods Thirty-two Sprague-Dawley (SD) rats were divided into four groups including a control group,a model group,a baclofen group,and a CGP35348 group.Liver fibrosis was then induced by carbon tetrachloride (CCl4).Baclofen and CGP35348 treatment were carried out after the formation of liver fibrosis,followed by complete extraction of the eyeball to obtain blood sample to test liver function.Liver tissue specimens were cut and stored for histological staining,histochemistry,real-time polymerase chain-reaction (RT-PCR),and western blot analysis.Results Histological staining indicated that the degree of liver fibrosis was more severe in the CGP35348 group than in the baclofen group (P<0.001).The levels of alanine transaminase (ALT),aspartate aminotransferase (AST),gamma-glutamyl transferase (GGT),total bilirubin (TBil),and direct bilirubin (DBil) were significantly lower in the baclofen group than in the CGP35348 group (P<0.01).The levels of ALT,AST,GGT,TBil,and DBil were significantly higher in the CGP35348 group than in the model group (P<0.05).Immunofluorescence results show that the hepatic cell migration was inhibited in the baclofen group.Western blot results showed that the expression levels of α-SMA protein were significantly lowered in the baclofen group when compared to that of the CGP35348 group and model group (P<0.01).Conclusion GABAB receptor might play a role in the liver protection by inhibition of migration of hepatic cells in liver fibrosis.Further studies into the mechanism behind this function are further needed and may be a potential source of future anti-fibrotic treatment.

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