1.Expression and clinical significance of microRNA-204 in human renal clear cell carcinoma
Linchao ZHANG ; Junfeng ZHAO ; Jijian SUN
Chongqing Medicine 2015;(15):2034-2036
Objective To inveatigate the expression and clinical significance of microRNA‐204(miR‐204) in human renal clear cell carcinoma(RCCC) .Methods 65 cases of resected RCCC tissue and corresponding normal tumor‐adjacent tissue were col‐lected and detected the expression level of miR‐204 by using qRT‐PCR .The correlation between the miR‐204 expression level and clilicalpathological features was statistically analyzed .The artificial miR‐204 mimics were adopted to stranfect into Caki‐1 cells ,then the Western blot was used to detect the expression of Bcl‐2 a as the downstream potential targets of miR‐204 and SIRT1 mRNA and protein .Results The expression level of miR‐204 in the RCCC tissue was significantly lowere than that in the normal tumor‐adjacent tissues (P<0 .05);the low expression of miR‐204 was significantly associated with the tumor size (>3 cm ,P<0 .05) , lymph node metastasis(P<0 .05) and advanced TNM stage(Ⅲ + Ⅳ ,P<0 .05) .Both the mRNA and protin expression in RCCC Caki‐1 cells were down‐regulated after transfection .Conclusion The expression of miR‐204 is down-regulated in the RCCC tissue and is related to the malignant clinicopathological features ,and miR‐204 may suppress the RCCC genesis and development through down‐regulating the expression of Bcl‐2 and SIRT1 .
2.Effect of preoperative neutrophil/lymphocyte ratio on the prognosis of patients with prostate cancer
Jijian SUN ; Junfeng ZHAO ; Linchao ZHANG
Journal of Chinese Physician 2015;17(11):1640-1642,1646
Objective To explore effect of preoperative neutrophil/lymphocyte ratio (NLR) on the prognosis of patients with prostate cancer.Methods A total of 51 cases died included in our hospital as the died group, also included 124 patients survival patients served as survival group.Preoperative total prostate specific antigen (tPSA), free prostate specific antigen (fPSA), albumin, white blood cell (WBC), neutrophil count, lymphocyte count, Gleason score, and NLR were collected.Receiver operating characteristic (ROC) curve was drawn.The function of the index on prognosis was evaluated.Results The average age was 74.0 (9) years in the died group, and 73.6 (11.0) years in the survival group.Gleason score was 8 ~ 10.The patients were followed up for 10 ~ 36 months with a median follow-up time of 23 months.No significant differences in albumin and Gleason score were found between death and survival groups (P >0.05).However, the other major indexes had significant differences between two groups (P < 0.01).NLR area under curve (AUC) was 0.835.The NLR boundary value was 2.84, which was higher than the critical value of patients with poor prognosis.Conclusions NLR can predict the prognosis of patients with malignant prostate cancer as auxiliary index.Patients with NLR > 2.83 had poor prognosis.
3.Effect of CIK cells immunotherapy on immunological function of patients with metastatic renal cell carcinoma
Hongquan CUI ; Junfeng ZHAO ; Baodong LI ; Linchao ZHANG
China Modern Doctor 2014;(29):132-133,136
Objective To study the effect of cytokine-induced killer(CIK) cells immunotherapy on immunological func-tion of patients with metastatic renal cell carcinoma.Methods A total of 70 patients with metastatic renal cell carcino-ma were randomly divided into 2 groups, 35 cases of the treatment group were added with CIK cells, while the control group was given IL-2 and IFN-α, detected the immunological function of 70 cases including the levels of CD3+, CD4+, CD8+T cells, ratio of CD4+/CD8+, one week before therapy of CIK cells and one month week after treatment. Results The levels of CD3+、CD4+cells, ratio of CD4+/CD8+in peripheral blood of patients with renal cancer of two groups were much higher than those before therapy (P<0.05), the levels CD8+ was lower after treatment, there were significant changes (P<0.05). Conclusion CIK cells immunotherapy can enhance the immunological function of patients with meta-static renal cell carcinoma.
4.Oligomerization triggered by foldon to enhance the catalytic efficiency of feruloyl esterase.
Lei ZHANG ; Linchao LEI ; Guangya ZHANG ; Xialan LI
Chinese Journal of Biotechnology 2019;35(5):816-826
A new method to express oligomerized feruloyl esterase (FAE) in Pichia pastoris GS115 to improve the catalytic efficiency was developed. It was realized by fusing the foldon domain at the C-terminus of FAE, and the fusion protein was purified by histidine tag. Fusion of the feruloyl esterase with the foldon domain resulted spontaneously forming a trimer FAE to improve the catalytic performance. The oligomerized FAE and monomeric FAE were obtained by purification. The apparent molecular weight of the oligomerized FAE was about 110 kDa, while the monomeric FAE about 40 kDa, and the optimum temperature of the oligomerized FAE was 50 °C, which is the same as the monomeric one. The optimal pH of the oligomerized FAE is 5.0, while the optimal pH of the monomer FAE is 6.0. When compared with the monomeric ones, the catalytic efficiency (kcat/Km) of the oligomerized FAE increased 7.57-folds. The catalytic constant (kcat) of the oligomerized FAE increased 3.42-folds. The oligomerized FAE induced by foldon have advantages in the catalytic performances, which represents a simple and effective enzyme-engineering tool. The method proposed here for improving the catalytic efficiency of FAE would have great potentials for improving the catalytic efficiency of other enzymes.
Carboxylic Ester Hydrolases
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metabolism
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Catalysis
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Molecular Weight
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Pichia
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genetics
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metabolism
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Polymerization
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Protein Engineering
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Substrate Specificity