Objective To evaluate the clinical effect of levocarnitine in the treatment of continuous blood purification and its effect on matrix metalloproteinase-2(MMP-2),interleukin-18(IL-18)and heart function.Methods 118 patients with continuous blood purification were randomly divided into the control group(n=59)and treatment group group(n=59).Two groups were treated with hemodialysis treatment.Control group was given folic acid,vitamin B and alpha keto acid,treatment group intravenous given levocarnitine injection 20mg/kg+0.9％NaCl 20mL on the basis of control group.A cycle of treatment was 28 days,and treated for 3 cycles.Serum levels of MMP-2 and IL-18,heart function,clinical effect and adverse drug reaction were compared between the two groups.Results After treatment,the total effective rate in control group was 84.74％,lower than 94.92％in treatment group(P<0.05).After treatment,serum MMP-2,IL-18,left atrial diameter(LAD),left ventricular end diastolic diameter(LVDd),left ventricular posterior wall thickness(LVPWT),ventricular septal thickness(IVST),left ventricular volume index(LVMI)in two groups were lower than pre-treatment(P<0.05),and the indexes in treatment group were more lower than control group(P<0.05).The E/A and left ventricular ejection fraction(LVEF)in two groups were all increased after treatment,and those in treatment group were higher than control group,the differences were significant(P<0.05).There was no significant in the incidence rate of adverse drug reactions between two groups.Conclusion Levocarnitine in the treatment of continuous blood purification was effective,and significantly reduce the serum MMP-2,IL-18 level and improve the heart function.

Objective To study the immuno-protective effect against Schistosoma japonicum challenge of the positive monoclonal phages which were screened from the 12 mers-phage random peptide library by the new model rabbit serum. Methods The new model was established by injecting the Schistosoma japonicum infection rabbits with inhibitor of phenol oxidose. Positive clones immunoscreened with the new model rabbit sera were absorbed by SEA immune rabbit sera, and 14 clones selected randomly from them were compared and their antigenic ability was identified by ELISA. The two best positive monoclonal phages (No.8, No.13) recognized by the new model rabbit sera were selected to immunize Kunming mice by subcutaneously injecting 1?10~15 pfu positive phages at 0, 2nd, 4th week respectively. After 4 weeks of the last immunity,each mouse was challenged with 40?1 S.japonicum cercariae. All mice were sacrificed after 42 days and the reduction rates of adult worms and the liver eggs were investigated. Results The positive phage clones after immune absorption were weakly recognized by the SEA immune rabbit sera. The 14 monoclonal phages were recognized by the rabbit sera of the new model and the normal model. Especially No.8, No.13 were strongly recognized by the rabbit sera of the new model,while weakly recognized by the SEA immune rabbit sera. The reduction rates of adult worms and liver eggs induced by the monoclonal phage No.13, the monoclonal phage No.8 and the original peptide library were 35.81% and 63.32%, 32.09% and 52.02%, 14.90% and 30.64%, respectively. Conclusion Most clones of simulated epitopes of SEA can be removed by absorbing positive clones with SEA immune rabbit serum .The 14 monoclonal phages from the new model contain the simulated S.japonicum epitope. The two monoclonal phages have higher reduction rates of adult worms and eggs than original 12 mers-phage random peptide library and the positive polyclonal phages.[

Objective To screen the 12 mers-phage random peptide library using the serum from the new model rabbit and to identify the immuno-protection of the positive phages. The new model infected with Schistosoma japonicum was proved that has a high protection against the challenge infection. Methods After being absorbed by E.coli antibody, the serum of the new model rabbit was used to screen the peptide library. Through three rounds of biopaning and enriching, lots of positive phages were obtained and their antigenic ability was tested. Every mouse was immunized by subcutaneously injecting 1?10 14 pfu positive phages from the new model rabbit serum respectively at 0-2-4 th week. After 4 weeks of the last immunization, the challenge infection was performed. At the same time, several control groups including the group immunized with the phages from the rabbit serum of the normal model infected with Schistosoma japonicum, the group immunized with the original 12 mers-phage random peptide library and the control group of challenge infection were arranged. Results ①The positive clones of phage(1?10 14) from the new model rabbit serum were strongly recognized by the rabbit serum of the new model, weakly recognized by the rabbit serum of the normal model infected with Schistosoma japonicum,but not recognized by the serum of healthy rabbit. ②The reduction rate of adult worms and liver eggs induced by phages screened with the rabbit serum of the new model group and the nomal model group and that induced by the original peptide library were respectively 27 2% and 38 8 %, 17 8% and 35 0%, 4 5% and 6 0% Conclusion The new model group obtained a higher reduction rate of adult worms than the nomal model group (P

Objective To observe the influencing of Yiqi Huayu Jiedu Prescription on the growth of HepG2 nude mice transplantation tumor and the expression of related factors of vascular mimicry. Methods Models of transplanted tumors, which were made by HepG2 cells in nude mice, were randomly divided into 7 groups, Yiqi Huayu Jiedu Prescription group, Astragali Radix group, Curcumae Rhizoma group, Paridis Rhizoma group, Gecko group, cis-platinum group, and model group. Except for the model group, the rest groups were given relevant medicine for intervention. 21 days later. HIF-1α, MMP-2, MMP-9, and E-cad were detected by immunohistochemistry, and Twist1 and Bcl-2 were detected by fluorescence quantitative PCR. Results Compared with the model group, tumor volume in the rest groups decreased (P<0.05), and the effect in the Yiqi Huayu Jiedu Prescription group was more obvious than the Astragali Radix group, Paridis Rhizoma group and Gecko group (P<0.05);The expression of vasculogenic mimicry structure was rare in each group, and the model group and cis-platinum group were the most obvious;Except for the Astragali Radix group, the expressions of HIF-1α, MMP-2, and MMP-9 showed statistical significance compared with model group (P<0.05);The expression of E-cad in the Yiqi Huayu Jiedu Prescription group and Astragali Radix group showed statistical significance (P<0.05);The expression of Bcl-2 in the Yiqi Huayu Jiedu prescription group, Paridis Rhizoma group, and Gecko group decreased significantly compared with the model group (P<0.05);The expression of Bcl-2 in the Yiqi Huayu Jiedu prescription group was much better than the other groups (P<0.05);The expression of Twist1 showed statistical significance in the Yiqi Huayu Jiedu Prescription group, Curcumae Rhizoma group, Paridis Rhizoma group, and cis-platinum group (P<0.05). Conclusion Yiqi Huayu Jiedu Prescription can reduce expression of HIF-1α, Twist1, Bcl-2, MMP-2, and MMP-9, and increase expression of E-cad, thereby inhibiting the formation of vascular mimicry.

Objective:Meta analysis was used to evaluate the diagnostic value of miRNA-25 in different malignant tumors in Chinese population.Methods:The research on the diagnostic value of miRNA-25 in China national knowledge internet (CKNI), VIP, Wanfang, PubMed and EMBase was searched. The publication time was from the establishment of database to October 2019. After the data was extracted, the Meta.DiSc1.4 software was used for preliminary statistical analysis. The Review Manager 5.3's QUADAS-2 tool was used for quality control of the incoming literature. The STATA 15.0 software was used to analyze whether there was publication bias.Results:A total of 10 articles were included in the study, including 792 cases in the case group and 715 cases in the control group. The combined sensitivity, combined specificity, combined specificity, combined positive likehood ratio(PLR), negative likehood ratio (NLR) were 0.71(95% CI: 0.69-0.74), 0.84(95% CI: 0.81-0.86), 4.29(95% CI: 3.39-5.42), 0.35(95% CI: 0.31-0.40), respectively. Diagnostic odds rate (DOR) was 13.09(95% CI: 9.64-17.78), and the area under curve (AUC) was 0.80(95% CI: 0.77-0.84). Subgroup analysis showed that the diagnostic value of miRNA-25 for different malignant tumors suggested that the lung cancer group had the highest diagnostic value and specificity (DOR=16.418, SPE=0.863). Funnel plot indicated that no significant publication bias was found ( P=0.21). Conclusions:miRNA-25 is highly sensitive and specific to different malignancies, especially lung cancer, and can be used as a potential screening biomarker to assist diagnosis.

Objective:To study the biological behavior of nasopharyngeal carcinoma stem cells and to explore the activation of Ras signaling pathway regulated by CD44.Methods:CNE2-SC and 5-8F-SC were nasopharyngeal carcinoma stem cells and obtained by serum-free suspension culture. Cell counting kit-8 (CCK-8) assay, colony formation assay, Transwell migration assay, cell adhesion array were used to investigate the growth, proliferation, migration and adhesion of nasopharyngeal carcinoma stem cells. Western blot test was used to detect the expressions of Ras signaling pathway related proteins and siRNA-mediated interference was used to determine the activation of Ras signaling pathway regulated by CD44.Results:The growth rates of CNE2-SC and 5-8F-SC cells were significantly lower than those of nasopharyngeal carcinoma cells at 24, 48 and 72 hours after inoculation ( P<0.05). After 14 days of implantation, the colony formation rates of CNE2-SC (44.5±1.9)% and 5-8F-SC (47.4±1.8)% were higher than those of CNE2 (34.9±1.5)% and 5-8F (37.2±1.7)%, respectively( P<0.01). The migration cell number of CNE2-SC was (87.6±7.8), 3.97 times higher than that of CNE2 ( P<0.01). The migration cell number of 5-8F-SC was (67.2±5.7), 3.07 times higher than 5-8F ( P<0.01). The adhesion rates of CNE2-SC and CNE2 cells were (42.1±7.6)% and (8.9±2.0)%, respectively at 3 hours after inoculation and were (82.4±5.0)% and (12.1±2.2)% at 6 hours after inoculation, respectively. The adhesion rate of CNE2-SC cells was higher than that of CNE2 cells (all P<0.01). The adhesion rates of 5-8F-SC and 5-8F cells were (53.6±6.1)% and (7.3±1.5)% at 3 hours after inoculation, and (90.7±3.6)% and (11.0±1.2)% at 6 hours after inoculation, respectively. The adhesion rate of 5-8F-SC cells was higher than that of 5-8F cells ( P<0.01). The expression levels of CD44, Ras and N-cadherin were significantly higher, while phosphatase and tensin homolog deleted on chromosome 10 (PTEN), E-cadherin in nasopharyngeal carcinoma stem cells were lower than those of the nasopharyngeal carcinoma cells. Furthermore, the levels of phosphorylated mitogen extracellular kinase1/2 (p-MEK1/2) and phosphorylated extracellular signal-regulated protein kinase1/2 (p-ERK1/2)were significantly increased in nasopharyngeal carcinoma stem cells ( P<0.01). Correlation analysis showed that the protein expression levels of CD44 was highly positively correlated with RAS in nasopharyngeal carcinoma stem cells( r=0.985, P=0.002; r=0.962, P=0.038). Deletion of CD44 in CNE2-SC decreased the expression levels of HER-2, Ras and p-ERK1/2, p-Akt and phosphorylated protein kinase C-δ(p-PKCδ) ( P<0.01). Conclusions:Despite compare to the nasopharyngeal carcinoma cell, nasopharyngeal carcinoma stem cells grows at a relatively slow rate, the capacities of clone formation, migration, adhesion are promoted. This may be related to the CD44-regulated abnormal activation of Ras signaling pathway.

Objective:To study the biological behavior of nasopharyngeal carcinoma stem cells and to explore the activation of Ras signaling pathway regulated by CD44.Methods:CNE2-SC and 5-8F-SC were nasopharyngeal carcinoma stem cells and obtained by serum-free suspension culture. Cell counting kit-8 (CCK-8) assay, colony formation assay, Transwell migration assay, cell adhesion array were used to investigate the growth, proliferation, migration and adhesion of nasopharyngeal carcinoma stem cells. Western blot test was used to detect the expressions of Ras signaling pathway related proteins and siRNA-mediated interference was used to determine the activation of Ras signaling pathway regulated by CD44.Results:The growth rates of CNE2-SC and 5-8F-SC cells were significantly lower than those of nasopharyngeal carcinoma cells at 24, 48 and 72 hours after inoculation ( P<0.05). After 14 days of implantation, the colony formation rates of CNE2-SC (44.5±1.9)% and 5-8F-SC (47.4±1.8)% were higher than those of CNE2 (34.9±1.5)% and 5-8F (37.2±1.7)%, respectively( P<0.01). The migration cell number of CNE2-SC was (87.6±7.8), 3.97 times higher than that of CNE2 ( P<0.01). The migration cell number of 5-8F-SC was (67.2±5.7), 3.07 times higher than 5-8F ( P<0.01). The adhesion rates of CNE2-SC and CNE2 cells were (42.1±7.6)% and (8.9±2.0)%, respectively at 3 hours after inoculation and were (82.4±5.0)% and (12.1±2.2)% at 6 hours after inoculation, respectively. The adhesion rate of CNE2-SC cells was higher than that of CNE2 cells (all P<0.01). The adhesion rates of 5-8F-SC and 5-8F cells were (53.6±6.1)% and (7.3±1.5)% at 3 hours after inoculation, and (90.7±3.6)% and (11.0±1.2)% at 6 hours after inoculation, respectively. The adhesion rate of 5-8F-SC cells was higher than that of 5-8F cells ( P<0.01). The expression levels of CD44, Ras and N-cadherin were significantly higher, while phosphatase and tensin homolog deleted on chromosome 10 (PTEN), E-cadherin in nasopharyngeal carcinoma stem cells were lower than those of the nasopharyngeal carcinoma cells. Furthermore, the levels of phosphorylated mitogen extracellular kinase1/2 (p-MEK1/2) and phosphorylated extracellular signal-regulated protein kinase1/2 (p-ERK1/2)were significantly increased in nasopharyngeal carcinoma stem cells ( P<0.01). Correlation analysis showed that the protein expression levels of CD44 was highly positively correlated with RAS in nasopharyngeal carcinoma stem cells( r=0.985, P=0.002; r=0.962, P=0.038). Deletion of CD44 in CNE2-SC decreased the expression levels of HER-2, Ras and p-ERK1/2, p-Akt and phosphorylated protein kinase C-δ(p-PKCδ) ( P<0.01). Conclusions:Despite compare to the nasopharyngeal carcinoma cell, nasopharyngeal carcinoma stem cells grows at a relatively slow rate, the capacities of clone formation, migration, adhesion are promoted. This may be related to the CD44-regulated abnormal activation of Ras signaling pathway.

Objective To study the effect of medicated serum of Jianpi Yiqi Huayu Jiedu Formula (JY-HJF),a TCMcompound formulated according to TCMprinciple of tonifying spleen,replenishing qi,re-moving blood stasis and detoxification,on proliferation,cycles,apoptosis of HCT116 colorectal cancer cells and on the protein expression of Caspase-3,Bcl-xL and Bax of HCT116 cells.Methods After drug-contained serum (10%,15%,20%)treated HCT116 cells for 12,24,48 and 72 h,the prolifera-tion was detected by MTT method;after treated for 24 or 48 h,the cell cycle and apoptosis rate were de-tected by propidium iodide (PI)staining.The protein expression level of Caspase 3,Bcl-xL and Bax in HCT116 cells were detected by Western blot.The control group was treated by blank serum and 10% fe-tal bovine serum (FBS).Results Compared with the same concentration of blank serum group,the cell proliferation was inhibited significantly in 15% and 20% medicated serum group after 12 h (P <0.05), and the inhibitory effect showed time-and concentration-dependence with the highest at 72 h (P <0. 05).In 10% and 15% medicated serum group,cells in G2 phase decreased,but the G1-phase cells and S-phase cells increased;cells in G1 phase and S phase increased and those in G2 phase decreased in 10% medicated serum group with the remarkable results at 48 h.It was suggested that the medicated ser-um could blocked cell cycles,with the positive correlation with time.Compared with the blank serum groups at the same concentration,after treated 24 and 48 h,apoptosis rate of HCT116 cells increased, and had the positive correlation with time and concentration.Compared with the blank serum group,the expression of Bax protein was upregulated and that of Caspase-3 and Bcl-xL were downregulated in 15%medicated serum group (P <0.05).Conclusion The effect of Jianpi Yiqi Huayu Jiedu Formula on in-hibiting proliferation and inducing apoptosis may be via the way of blocking HCT116 cells cycles and of upregulating Bax protein expression with downregulating Caspase-3 protein and Bcl-xL protein expression.

Objective To establish a simple,feasible and reliable orthotopic implantation and metastasis model of human colon cancer in nude mice cecum mucosa.Methods Tumor cell line HCT116,derived from human colon cancer,was injected into colon cecum mucosa of nude mice to develop colorectal cancer model.HE staining and human carcinoma antigen (CEA) immunohistochemistry were performed to detect tumor in situ and metastasis condition in abdominal lymph nodes,the liver and the lungs.Results On the 2nd week after inoculation,90％ of the nude mice developed cecum tumor of 2-5 mm in diameter in situ.Abdominal lymph node metastasis was observed on the 4th week and present in all mice at week 12.Liver and lung metastasis started to appear at a percentage of 60％ on the 10th week and 39％ on the 12th week.Conclusion Human colon cancer in situ could be successfully established by inoculating HCT116 into cecum mucosa of nude mice to mimic the clinical development and growth of colon cancer.This method proves to be simple with a high tumor-bearing rate.It could be used as a reliable and ootimal animal model for in-depth basic research about the mechanism of colon cancer metastasis.

Cancer is a prevalent and challenging disease that is difficult to treat.Western medicine recognizes the disruption of the immune and inflammatory microenvironment as a crucial factor in the development and progression of cancer.According to traditional Chinese medicine,cancer falls into the categories of"accumulation"and"abnormal masses".Based on the attributes of cold and heat in the overall and local conditions as well as the characteristic of the struggle between the vital qi and pathogenic factors at different stages of this disease,this article proposes the concept of"body coldness and tumor heat"to describe the pathogenesis of cancer formation and progression.This pathogenesis aligns with the disruption of the immune and inflammatory microenvironment in modern medical oncology.The article suggests a treatment approach that focuses on balancing cold and heat,nourishing and protecting yang qi,warming the kidneys,and strengthening the spleen to address the"coldness of the entire body".This approach also involves promoting qi circulation,eliminating dampness,phlegm,and stasis,and detoxifying and dispersing nodules to address the"heat in the tumor locally".By addressing deficiencies,eliminating pathogenic factors,and promoting circulation to alleviate stagnation,the aim is to restore the balance of yin and yang and improve the complex state of"coldness of the entire body"and"heat in the tumor locally".These interventions can ameliorate the disorder in the microenvironment and enhance clinical efficacy.