1.Genotyping of human platelet antigen system 5 by PCR restriction fragment length polymorphism
Zhenyu LU ; Dazhuang LIU ; Ziyan ZHU
Chinese Journal of Laboratory Medicine 2001;0(04):-
Objective To develop a polymerase chain reaction restriction fragment length polymorphism (PCR RFLP) method using designed primers for determining the genotype of humen platelet antigens (HPA)5 system. Methods HPA 5 system of 25 healthy blood donors were genotyped using PCR RFLP method. The results obtained by PCR RFLP were compared with those determined by allele specific oligonucleotid hybridization (PCR ASO). Results The results of HPA 5 system obtained by PCR RFLP in 25 health donors were as follows: 24 of aa, 1 of ab and 0 of bb. All were in good agreement with those determined by PCR ASO. Conclusions Because PCR RFLP method is plain, fast and reliable for HPA 5 system genotyping, it is suitable for the diagnosis and therapy of neonatal alloimmune thrombocytopenia, posttransfusion purpura, platelet transfusion refractoriness and so on..
2.Effects of metabolic abnormality on the incidence of colorectal cancer
Dazhuang MIAO ; Zheng LIU ; Xishan WANG
Chinese Journal of Digestive Surgery 2014;13(12):995-998
Metabolic abnormality is correlated with the incidence of colorectal cancer.Metabolic syndrome is consisting of abdominal obesity,dyslipidemia,high blood pressure,sugar metabolic abnormality,and all of these factors are associated with colorectal cancer.Abdominal obesity and sugar metabolic abnormality may be the primary factors causing rectal cancer.Visceral fat is physiologically more active than subcutaneous fat.Visceral fat can produce and secret hormones and cytokines,which are involved in inflammation and metabolism,therefore the amount of visceral fat may directly or indirectly related to the occurrence of colorectal cancer.Obesity acts as a risk factor for colorectal cancer by several mechanisms,including high blood insulin,insulin-like growth factor,and the change of the adipose cytokines concentrations.Metabolic biomarkers may not only provide clues of colorectal carcinogenesis from the point of view of etiology,but also can help to explore new obesity phenotypes that is relevant to incidence risk of colorectal cancer.
3.Study on B(A) phenotypes and identification of novel B(A)641 allele in chinese population
Zhonghui GUO ; Dong XIANG ; Ziyan ZHU ; Xi LIU ; Heping CHEN ; Dazhuang LIU
Chinese Journal of Blood Transfusion 1988;0(01):-
C mutation. All 8 samples displayed the B(A) phenotype. Their real genotypes were B(A)/O. Conclusion Three B(A) alleles in the Chinese Han population were detected. Two alleles,B(A)700,B(A)640 were reported previously. One novel allele B(A)641, was first identified in this study.
5.Influence of Paraquat Poisoning on the Expression of Toll-like Receptor 4 in Myocardial Damage Following in Mice and Its Significance
Xuesong DONG ; Rui WANG ; Xiaoyang XU ; Wei LIU ; Dazhuang SUN ; Zhi LIU
Journal of China Medical University 2015;(10):891-896,900
Objective To clarify the role and significance of Toll?like receptor 4(TLR4)in myocardial damage following paraquat(PQ)poisoning in mice. Methods Male wild type C57BL/6J mice(WT)and male TLR4 deficient mice(TLR4?ko)were divided into four groups in the study:(1)control group(WT mice,n=6);(2)TLR4?ko group(TLR4?ko mice,n=6);(3)WT+PQ group(WT mice,n=30);(4)TLR4?ko+PQ group (TLR4?ko mice,n=30). The mice in group 1 and group 2 were injected intraperitoneally with saline;mice in group 3 and group 4 were injected in?traperitoneally with 75 mg/kg of PQ. At 2 h,4 h,8 h,16 h and 24 h after PQ administration,6 mice of WT+PQ group and TLR4?ko+PQ group were euthanized and the heart tissue specimens were harvested. All the specimens were analysed by histology,while the expression of TLR4 mRNA was only detected in samples of WT+PQ mice. The specimens at 2 h,8 h and 24 h after PQ administration were used for cytokines detection for WT+PQ group and TLR4?ko+PQ group;in addition,Western blot analysis was performed for WT+PQ group. At 8 h after treatment,control mice and TLR4?ko mice were euthanized by the same method. Mice were anesthetized for cardiac geometry and functional assessment using a 2?D guide M?mode echocardiography at 8 h following injection of either PQ or saline. Results During myocardial damage due to PQ exposure in WT+PQ mice,obvious histopathological changes were observed,as well as a noticeable decrease of heart function and increased expressions of TNF?αand IL?1β. Compared with the WT mice,TNF?αand IL?1βprotein levels,changes in heart function and histopathological changes were significantly attenuated following PQ exposure in myocardial damage in TLR4?ko mice. Conclusion The TLR4gene is involved with in heart functional injury and histopathological changes in myocardial damage following PQ poisoning in mice,which may through the regulation of TNF?αand IL?1βexpression. Our findings indi?cate that TLR4 plays an important role in mediating myocardial injury due to PQ.
6.Establishment of platelet antigen panel and its application in the identification of platelet specific antibodies
Mingliang FENG ; Wei SHEN ; Zhonghui GUO ; Tong SHEN ; Biao YIN ; Jianlian WANG ; Sha JIN ; Dazhuang LIU
Chinese Journal of Laboratory Medicine 2009;32(2):162-164
Objective To establish the platelet antigen panel for identifying the specificity of platelet antibodies which cause platelet transfusion refractoriness and neonatal alloimmune thrombocytopenia and provide evidence for clinical therapy and platelet genotyping research.Methods Based on the frequency distribution of human platelet alloantigen (HPA)-1 to HPA-16 gene in China, the frequencies of HPA-1 to HPA-6,HPA-15 alleles in blood group O donors were genotyped by the polymerase chain reaction with sequence-specific primers (PCR-SSP) method, and suitable donors were chosen to establish platelet-specific antigen panel.Using the established platelet-specific antigen panel, the specificity of platelet antibodies caused by alloimmune reaction was identified by using simplified sensitized erythrocyte platelet serology assay (SEPSA).Results Eleven ptatelet donors with blood group O were chosen to establish platelet-specific antigen panel which can identify specificity of HPA-1 to HPA-6, HPA-15 antibodies.One case of HPA-4b (Penb) and two cases of HPA-15a (Govb) platelet specific antibodies were detected in 1 120 samples.Conclusion Identifying the specific platelet antibodies using platelet specific antigen panel has profound significance on increasing the safety and effectiveness of clinical platelet transfusion and prevention of neonatal alloimmune thrombocytopenia.
7.Expression profiles of CaMKIIδat different stages of osteoclast differenti-ation
Dazhuang LU ; Juanjuan LIU ; Mengchun QI ; Liming WEN ; Ren LI ; Hong SUN
Chinese Journal of Pathophysiology 2016;32(10):1870-1874
[ ABSTRACT] AIM:To study the expression profiles and the role of Ca 2+/calmodulin-dependent kinase II delta ( CaMKIIδ) during osteoclast differentiation .METHODS:Mouse RAW264.7 cells were induced by receptor activator of nuclear factor κB ligand ( RANKL) at 50μg/L for osteoclastogenesis .Tartrate-resistant acid phosphatase ( TRAP) staining and bone resorption lacunae examination were performed to verify osteoclast formation .The expression of CaMKIIδat mR-NA and protein levels was also determined by immunofluorescent cytochemistry , RT-qPCR and Western blot at days 0, 1, 3 and 5.RESULTS:TRAP positive multinuclear cells with bone resorption function were formed after 5 d of induction. The mRNA levels of CaMKIIδdetected by RT-qPCR were 1.028 ±0.041, 2.478 ±0.087, 10.524 ±1.284 and 42.914 ± 2.667 at days 0, 1, 3 and 5, respectively, while the protein levels of CaMKIIδ detected by Western blot were 0.762, 0.963, 1.802 and 3.136, respectively.The changes of protein level were also verified by immunofluorescence cytochemis -try, in which the fluorescence intensity increased in a time-dependent manner (P<0.05).CONCLUSION:The expres-sion of CaMKIIδincreases with the differentiation of osteoclasts .CaMKIIδmay play a key role in the osteoclastogenesis .
8.Effect of transanal excision in the treatment of rectal cancer.
Xishan WANG ; Zheng LIU ; Dazhuang MIAO
Chinese Journal of Gastrointestinal Surgery 2014;17(6):544-546
The surgical approach for rectal cancer includes trans-abdominal and transanal excision. Total mesorectal excision(TME) is the golden standard for surgical treatment. In the functional surgery era, more and more evidence shows that under strict indications, traditional abdominal radical surgery and transanal excision can achieve similar survival in patients with early stage cancer. However, the local recurrence rate of local resection was significantly higher compared to TME, suggesting strict patients selection for transanal excision. Preoperative accurate evaluation is critical in clinical practice.
Anal Canal
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surgery
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Digestive System Surgical Procedures
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methods
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Humans
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Rectal Neoplasms
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surgery
9.Preoperative prediction of lymphovascular and visceral pleural invasion of lung adenocarcinoma based on 18F-FDG PET radiomics
Xiaohui SUN ; Zhipeng LIU ; Dazhuang YANG ; Jun ZHANG
Chinese Journal of Nuclear Medicine and Molecular Imaging 2024;44(2):74-79
Objective:To evaluate the predictive value of 18F-FDG PET-based radiomics models for lymphovascular invasion (LVI) and visceral pleural invasion (VPI) in lung adenocarcinoma (LAC) prior to surgery. Methods:Eighty-seven patients with LAC (42 males, 45 females, age: (64.6±9.0) years; 90 lesions) pathologically confirmed in the Affiliated Taizhou People′s Hospital of Nanjing Medical University between August 2018 and August 2022 were retrospectively included. Based on the radiomics features extracted from PET images, the machine learning models were constructed by using the support vector machine (SVM), logical regression (LR), decision tree (DT), and K-nearest neighbor (KNN) algorithm. Stratified sampling (Python′s StratifiedkFold function) was employed to divide the data into training set and test set at a ratio of 8∶2. The model stability was assessed using the 50% discount cross-validation. The ROC curve was drawn, and the AUC was calculated to evaluate the value of radiomics models in predicting LVI and VPI in LAC. Delong test was used to compare AUCs of different models.Results:The radiomics models (SVM, LR, DT, KNN) based on PET images showed good predictive value for LVI and VPI in patients with LAC. For LVI, the AUCs were 0.91, 0.90, 0.91, 0.91 in the training set, and were 0.85, 0.87, 0.77, 0.78 in the test set; for VPI, the AUCs were 0.86, 0.86, 0.84, 0.81 in the training set, and were 0.82, 0.80, 0.69, 0.78 in the test set. The F1 scores of the SVM model were the best (0.59 and 0.66 for predicting LVI and VPI respectively). The Delong test showed that there were no significant differences in AUCs among the four models ( z values: from -1.46 to 1.71, all P>0.05). Conclusions:The machine learning models based on 18F-FDG PET radiomics features are effective in predicting LVI and VPI in patients with LAC prior to surgery. These models can assist clinicians in stratifying the risk of LAC and making informed clinical decisions. The SVM model has the best performance in predicting LVI and VPI.
10.Effect of transanal excision in the treatment of rectal cancer
Xishan WANG ; Zheng LIU ; Dazhuang MIAO
Chinese Journal of Gastrointestinal Surgery 2014;(6):544-546
The surgical approach for rectal cancer includes trans-abdominal and transanal excision. Total mesorectal excision (TME) is the golden standard for surgical treatment. In the functional surgery era, more and more evidence shows that under strict indications , traditional abdominal radical surgery and transanal excision can achieve similar survival in patients with early stage cancer. However , the local recurrence rate of local resection was significantly higher compared to TME, suggesting strict patients selection for transanal excision. Preoperative accurate evaluation is critical in clinical practice.