Objective:To explore the mechanism by which the extract of Hedyotis diffusa Willd regulates TGF-β/Smad signaling transduction pathway and affects proliferation,invasion of renal cancer cells and expressions of Th1/Th2 cytokines in mice with renal cancer.Methods:Using mouse renal cancer Renca cells as the research object,after intervention with extracts of Hedyotis diffusa Willd,impact on migration ability of mouse renal cancer cells was detected at the cellular level using scratch assay;observing effect of CCK-8 experiment on proliferation ability of mouse renal cancer cells.A mouse kidney cancer model was established by subcutaneous inoculation under the right armpit,and effect of extract of Hedyotis diffusa Willd on proliferation and invasion of renal carcinoma in mice was studied.HE staining was used to observe histomorphology of transplanted tumor mass in mice;Western blot was used to detected expressions of TGF-β/Smad pathway protein TGF-β,Smad2 and p-Smad2 in mouse transplanted tumor tissue;levels of IL-2 and IL-10 in mouse serum were detected by ELISA to investigate whether the extract of Hedyotis diffusa Willd has an inhibitory effect on Th1/Th2 renal carcinoma immune shift.Results:CCK-8 experiment showed that the extract of Hedyotis diffusa Willd inhibited pro-liferation ability of mouse renal cancer Renca cells(P<0.05);scratch test results showed that extract of Hedyotis diffusa Willd effec-tively inhibited the healing ability of mouse renal cancer Renca cells(P<0.05);by constructing a mouse kidney cancer model,it was found that extract of Hedyotis diffusa Willd could inhibit the growth of transplanted tumor volume in mice(P<0.05);HE staining revealed that tumor cells in model group were dense,with numerous mitotic figures and minimal necrosis;whereas in Hedyotis diffusa Willd group and pathway inhibitor group,there was increased necrosis of cancer cells,accompanied by karyopyknosis or lysis;Western blot showed that compared with model group,expression of TGF-β and p-Smad2 in extract of Hedyotis diffusa Willd group and pathway inhibitor group were decreased(P<0.001);ELISA results showed that compared with model group,serum IL-2 expression increased in both Hedyotis diffusa Willd group and pathway inhibitor group(P<0.000 1).IL-10 expression decreased in Hedyotis diffusa Willd group(P<0.05)and pathway inhibitor group(P>0.05).Conclusion:Extract of Hedyotis diffusa Willd may inhibit protein expressions in TGF-β/Smads signaling pathway,further regulate the secretion of Th1/Th2 cytokines,thereby inhibiting proliferation and migration of mouse renal cancer cells both in vitro and in vivo.

Objective:To explore the mechanism by which the extract of Hedyotis diffusa Willd regulates TGF-β/Smad signaling transduction pathway and affects proliferation,invasion of renal cancer cells and expressions of Th1/Th2 cytokines in mice with renal cancer.Methods:Using mouse renal cancer Renca cells as the research object,after intervention with extracts of Hedyotis diffusa Willd,impact on migration ability of mouse renal cancer cells was detected at the cellular level using scratch assay;observing effect of CCK-8 experiment on proliferation ability of mouse renal cancer cells.A mouse kidney cancer model was established by subcutaneous inoculation under the right armpit,and effect of extract of Hedyotis diffusa Willd on proliferation and invasion of renal carcinoma in mice was studied.HE staining was used to observe histomorphology of transplanted tumor mass in mice;Western blot was used to detected expressions of TGF-β/Smad pathway protein TGF-β,Smad2 and p-Smad2 in mouse transplanted tumor tissue;levels of IL-2 and IL-10 in mouse serum were detected by ELISA to investigate whether the extract of Hedyotis diffusa Willd has an inhibitory effect on Th1/Th2 renal carcinoma immune shift.Results:CCK-8 experiment showed that the extract of Hedyotis diffusa Willd inhibited pro-liferation ability of mouse renal cancer Renca cells(P<0.05);scratch test results showed that extract of Hedyotis diffusa Willd effec-tively inhibited the healing ability of mouse renal cancer Renca cells(P<0.05);by constructing a mouse kidney cancer model,it was found that extract of Hedyotis diffusa Willd could inhibit the growth of transplanted tumor volume in mice(P<0.05);HE staining revealed that tumor cells in model group were dense,with numerous mitotic figures and minimal necrosis;whereas in Hedyotis diffusa Willd group and pathway inhibitor group,there was increased necrosis of cancer cells,accompanied by karyopyknosis or lysis;Western blot showed that compared with model group,expression of TGF-β and p-Smad2 in extract of Hedyotis diffusa Willd group and pathway inhibitor group were decreased(P<0.001);ELISA results showed that compared with model group,serum IL-2 expression increased in both Hedyotis diffusa Willd group and pathway inhibitor group(P<0.000 1).IL-10 expression decreased in Hedyotis diffusa Willd group(P<0.05)and pathway inhibitor group(P>0.05).Conclusion:Extract of Hedyotis diffusa Willd may inhibit protein expressions in TGF-β/Smads signaling pathway,further regulate the secretion of Th1/Th2 cytokines,thereby inhibiting proliferation and migration of mouse renal cancer cells both in vitro and in vivo.

Objective To investigate the effect of Xuebijing injection on the inflammatory and coagulation response of complicated intra-abdominal infection by sepsis.Methods A retrospective study was conducted.A total of 274 patients with complicated intra-abdominal infection by sepsis admitted to Yidu Central Hospital of Weifang from June 2020 to July 2023 were enrolled.According to whether Xuebijing was used or not,the patients were divided into an observation group(141 cases)and a control group(133 cases).The patients in both groups were treated with conventional western medicine,and those in the observation group were additionally given Xuebijing(Xuebijing injection 100 mL in 0.9%NaCl solution 100 mL via intravenous infusion,twice per day for a continuous application of 5 days).The differences of clinical prognosis and the serum levels of inflammatory indicators[interleukin-6(IL-6),procalcitonin(PCT),C-reactive protein(CRP)and endotoxin]and coagulation indexes[D-dimer,fibrinogen(FIB),prothrombin time(PT)and activated partial thromboplastin time(APTT)before treatment and on the 2nd,3rd,5th day respectively after treatment were compared,and the clinical prognosis was observed.Results There were no significant differences in the indexes of inflammation and coagulation function between the two groups before treatment.With the extension of therapy duration,following treatment in two groups,the serum levels of inflammatory indicators and coagulation indexes(D-dimer,FIB)decreased,as did coagulation indexes(PT,APTT),reaching the lowest levels on the 5th day.The inflammatory markers and coagulation indexes of observation group were significantly lower than those of control group[IL-6(ng/L):18.89±8.22 vs.24.59±10.39,PCT(μg/L):0.30±0.16 vs.0.52±0.20,CRP(mg/L):13.13±6.54 vs.28.87±8.66,endotoxin(kEU/L):0.17±0.06 vs.0.22±0.08,D-dimer(mg/L):1.75±0.61 vs.1.96±0.77,FIB(g/L):2.50±0.78 vs.3.10±0.94,PT(s):14.16±1.90 vs.15.05±1.46,APTT(s):34.52±5.40 vs.36.21±4.72,both P<0.05].The intensive care unit(ICU)stay time and total hospitalization time were shortened in the observation group than those in the control group[ICU stay time(days):5.71±2.22 vs.6.86±2.99,hospitalization time(days):15.50±6.39 vs.17.02±5.70,both P<0.05];the mortality was significantly lower in the observation group than that in the control group[21.27%(30/141)vs.32.33%(43/133),P<0.05].Conclusion The treatment of complicated intra-abdominal infection by sepsis with Xuebijing injection while receiving conventional western medicine treatment can effectively decrease inflammatory and coagulation response,have high efficacy of inflammatory and coagulation co-treatment,and have some important clinical values.

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 ; metabolism ; Catalysis ; Molecular Weight ; Pichia ; genetics ; metabolism ; Polymerization ; Protein Engineering ; Substrate Specificity

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 .

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.

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.