BACKGROUND Hepatocellular carcinoma(HCC)is difficult to diagnose with poor therapeutic effect,high recurrence rate and has a low survival rate.The survival of patients with HCC is closely related to the stage of diagn...BACKGROUND Hepatocellular carcinoma(HCC)is difficult to diagnose with poor therapeutic effect,high recurrence rate and has a low survival rate.The survival of patients with HCC is closely related to the stage of diagnosis.At present,no specific serolo-gical indicator or method to predict HCC,early diagnosis of HCC remains a challenge,especially in China,where the situation is more severe.AIM To identify risk factors associated with HCC and establish a risk prediction model based on clinical characteristics and liver-related indicators.METHODS The clinical data of patients in the Affiliated Hospital of North Sichuan Medical College from 2016 to 2020 were collected,using a retrospective study method.The results of needle biopsy or surgical pathology were used as the grouping criteria for the experimental group and the control group in this study.Based on the time of admission,the cases were divided into training cohort(n=1739)and validation cohort(n=467).Using HCC as a dependent variable,the research indicators were incorporated into logistic univariate and multivariate analysis.An HCC risk prediction model,which was called NSMC-HCC model,was then established in training cohort and verified in validation cohort.RESULTS Logistic univariate analysis showed that,gender,age,alpha-fetoprotein,and protein induced by vitamin K absence or antagonist-II,gamma-glutamyl transferase,aspartate aminotransferase and hepatitis B surface antigen were risk factors for HCC,alanine aminotransferase,total bilirubin and total bile acid were protective factors for HCC.When the cut-off value of the NSMC-HCC model joint prediction was 0.22,the area under receiver operating characteristic curve(AUC)of NSMC-HCC model in HCC diagnosis was 0.960,with sensitivity 94.40%and specificity 95.35%in training cohort,and AUC was 0.966,with sensitivity 90.00%and specificity 94.20%in validation cohort.In early-stage HCC diagnosis,the AUC of NSMC-HCC model was 0.946,with sensitivity 85.93%and specificity 93.62%in training cohort,and AUC was 0.947,with sensitivity 89.10%and specificity 98.49%in validation cohort.CONCLUSION The newly NSMC-HCC model was an effective risk prediction model in HCC and early-stage HCC diagnosis.展开更多
Objective: The aim of our study was to investigate the effects of down-regulation Bmi-1 by RNA interference (RNAi) in T Lymphocytic leukemia Jurkat cells. Methods: Two complementary oligonucleotide strands were sy...Objective: The aim of our study was to investigate the effects of down-regulation Bmi-1 by RNA interference (RNAi) in T Lymphocytic leukemia Jurkat cells. Methods: Two complementary oligonucleotide strands were synthesized based on the siRNA sequence targeting Bmi-1 gene. After annealing, siRNA strands were recombined into the pRNAT- U6.2 vector, and then DNA sequencing was carded out following transformation and amplification. The recombinant was transfected into Jurkat cells with liposomes. Positive colonies were obtained through G418 selection. The mRNA and protein expressions of Bmi-1 were detected by RT-PCR and Western-blot, respectively. Effects of Bmi-1 silence on cell proliferation, cell cycle and cell aging of Jurkat cells were detected by M'l-r assay, flow cytometry, colony formation assay and SA-β-Gal staining, respectively. Results: The siRNA recombinant targeting Bmi.1 gene was successfully constructed. All three siRNA recombinants could significantly inhibit the expression of Bmi-l. The siRNA targeting 825nt-843nt (GACCAGACCACTACT GAAT) has the strongest inhibitory effect on Bmi-1 expression, with almost complete inhibition on Bmi-1 mRNA and protein expressions. Compared with the non-transfection group and the empty vector group, growth velocity and colony formation ability were significantly decreased, while the proportion of calls in G1 phase and the percentage of senile cells were signifi- cantly increased in highly transfected group (P 〈 0.05). Conclusion: Down-regulation Bmi-1 by RNA interference (RNAi) could significantly inhibit the growth of Jurkat cells in vitro.展开更多
Background Glanzmann thrombasthenia (GT) is an autosomal recessive bleeding disorder characterized by the tendency to hemorrhage and the inability of platelets to aggregate in response to agonists. GT is caused by a...Background Glanzmann thrombasthenia (GT) is an autosomal recessive bleeding disorder characterized by the tendency to hemorrhage and the inability of platelets to aggregate in response to agonists. GT is caused by a defect of the platelet glycoprotein lib/Ilia complex. The objective of this study was to describe the clinical features and the genetic cause of GT in a 6-year-old girl from south China. Methods A three-generation family was studied. The proband patient aged 6 years and her parents undertook examinations of platelet counts, blood film, bleeding time, platelet aggregation, and flow cytometry. All coding exons of the ITGA2B and ITGB3 genes were amplified by polymerase chain reaction (PCR), and direct sequencing was performed for mutational screening on the patient and normal controls consisted of 52 healthy blood donors. Reverse transcription PCR was conducted to test for exon skipping. Results The proposita patient showed dispersing platelets, prolonged bleeding time, and severely reduced platelet aggregation in response to the physiological agonists adenosine diphosphate (ADP), epinephrine, collagen, and ristocetin. Flow cytometric measurements showed that the contents of allb and 133 were significantly decreased. Sequencing results demonstrated two different types of heterozygous mutations existed in the allb gene (c.2930delG and IVS15-1delG). The compound mutations were also confirmed in the patient's mother and father separately. Conclusions The allbβ3 deficiency of the proband was caused by two compound ITGA2B mutations, which were first reported in Chinese GT patients. The IVS15-1delG was first confirmed to cause an exon skipping.展开更多
文摘BACKGROUND Hepatocellular carcinoma(HCC)is difficult to diagnose with poor therapeutic effect,high recurrence rate and has a low survival rate.The survival of patients with HCC is closely related to the stage of diagnosis.At present,no specific serolo-gical indicator or method to predict HCC,early diagnosis of HCC remains a challenge,especially in China,where the situation is more severe.AIM To identify risk factors associated with HCC and establish a risk prediction model based on clinical characteristics and liver-related indicators.METHODS The clinical data of patients in the Affiliated Hospital of North Sichuan Medical College from 2016 to 2020 were collected,using a retrospective study method.The results of needle biopsy or surgical pathology were used as the grouping criteria for the experimental group and the control group in this study.Based on the time of admission,the cases were divided into training cohort(n=1739)and validation cohort(n=467).Using HCC as a dependent variable,the research indicators were incorporated into logistic univariate and multivariate analysis.An HCC risk prediction model,which was called NSMC-HCC model,was then established in training cohort and verified in validation cohort.RESULTS Logistic univariate analysis showed that,gender,age,alpha-fetoprotein,and protein induced by vitamin K absence or antagonist-II,gamma-glutamyl transferase,aspartate aminotransferase and hepatitis B surface antigen were risk factors for HCC,alanine aminotransferase,total bilirubin and total bile acid were protective factors for HCC.When the cut-off value of the NSMC-HCC model joint prediction was 0.22,the area under receiver operating characteristic curve(AUC)of NSMC-HCC model in HCC diagnosis was 0.960,with sensitivity 94.40%and specificity 95.35%in training cohort,and AUC was 0.966,with sensitivity 90.00%and specificity 94.20%in validation cohort.In early-stage HCC diagnosis,the AUC of NSMC-HCC model was 0.946,with sensitivity 85.93%and specificity 93.62%in training cohort,and AUC was 0.947,with sensitivity 89.10%and specificity 98.49%in validation cohort.CONCLUSION The newly NSMC-HCC model was an effective risk prediction model in HCC and early-stage HCC diagnosis.
文摘Objective: The aim of our study was to investigate the effects of down-regulation Bmi-1 by RNA interference (RNAi) in T Lymphocytic leukemia Jurkat cells. Methods: Two complementary oligonucleotide strands were synthesized based on the siRNA sequence targeting Bmi-1 gene. After annealing, siRNA strands were recombined into the pRNAT- U6.2 vector, and then DNA sequencing was carded out following transformation and amplification. The recombinant was transfected into Jurkat cells with liposomes. Positive colonies were obtained through G418 selection. The mRNA and protein expressions of Bmi-1 were detected by RT-PCR and Western-blot, respectively. Effects of Bmi-1 silence on cell proliferation, cell cycle and cell aging of Jurkat cells were detected by M'l-r assay, flow cytometry, colony formation assay and SA-β-Gal staining, respectively. Results: The siRNA recombinant targeting Bmi.1 gene was successfully constructed. All three siRNA recombinants could significantly inhibit the expression of Bmi-l. The siRNA targeting 825nt-843nt (GACCAGACCACTACT GAAT) has the strongest inhibitory effect on Bmi-1 expression, with almost complete inhibition on Bmi-1 mRNA and protein expressions. Compared with the non-transfection group and the empty vector group, growth velocity and colony formation ability were significantly decreased, while the proportion of calls in G1 phase and the percentage of senile cells were signifi- cantly increased in highly transfected group (P 〈 0.05). Conclusion: Down-regulation Bmi-1 by RNA interference (RNAi) could significantly inhibit the growth of Jurkat cells in vitro.
文摘Background Glanzmann thrombasthenia (GT) is an autosomal recessive bleeding disorder characterized by the tendency to hemorrhage and the inability of platelets to aggregate in response to agonists. GT is caused by a defect of the platelet glycoprotein lib/Ilia complex. The objective of this study was to describe the clinical features and the genetic cause of GT in a 6-year-old girl from south China. Methods A three-generation family was studied. The proband patient aged 6 years and her parents undertook examinations of platelet counts, blood film, bleeding time, platelet aggregation, and flow cytometry. All coding exons of the ITGA2B and ITGB3 genes were amplified by polymerase chain reaction (PCR), and direct sequencing was performed for mutational screening on the patient and normal controls consisted of 52 healthy blood donors. Reverse transcription PCR was conducted to test for exon skipping. Results The proposita patient showed dispersing platelets, prolonged bleeding time, and severely reduced platelet aggregation in response to the physiological agonists adenosine diphosphate (ADP), epinephrine, collagen, and ristocetin. Flow cytometric measurements showed that the contents of allb and 133 were significantly decreased. Sequencing results demonstrated two different types of heterozygous mutations existed in the allb gene (c.2930delG and IVS15-1delG). The compound mutations were also confirmed in the patient's mother and father separately. Conclusions The allbβ3 deficiency of the proband was caused by two compound ITGA2B mutations, which were first reported in Chinese GT patients. The IVS15-1delG was first confirmed to cause an exon skipping.