Objective:To review and analyze the clinical diagnosis and treatment of renal Ewing's sar-coma with venous tumor embolus,to follow up the survival and prognosis of the patients,and to provide help for the diagnosis and treatment of the disease.Methods:Clinical data(including general data,sur-gical data and postoperative pathological data)of patients diagnosed with renal Ewing's sarcoma with ve-nous tumor embolus in Peking University Third Hospital from June 2016 to June 2022 were collected,and the prognosis of the patients was followed up to analyze the influence of diagnosis and treatment process on the prognosis of the disease.Results:There were 6 patients,including 1 male and 5 females.There were 4 cases of left renal tumor and 2 cases of right renal tumor.The median age at diagnosis was 28 years(16-52 years).The imaging findings were all exogenous tumors with internal necrotic tissue and hemorrhage.The mean maximum tumor diameter was 12.6 cm,and the mean tumor thrombus length was 7.8 cm.Four patients underwent open surgery and 2 patients underwent laparoscopic surgery.The post-operative pathological results were renal Ewing sarcoma.Immunohistochemical results showed 3 cases of CD99(+),2 cases of FLI-1(+),and 1 case of CD99,FLI-1(-).3 patients received chemothera-py(cyclophosphamide,doxorubicin,vincristine/ifosfamide,etoposide),1 case received chemotherapy combined with radiotherapy,and 2 cases received no adjuvant therapy.The mean overall survival(OS)of the 6 patients was 37 months,and the mean OS of the 4 patients(47 months)who received chemo-therapy was significantly higher than that of the 2 patients(16 months)who did not receive chemotherapy(P=0.031).Conclusion:Renal Ewing's sarcoma with venous tumor embolus is rare in clinic,and it is common in young female patients.The operation is difficult and the prognosis is poor.Surgical resection,adjuvant radiotherapy and chemotherapy can improve the overall survival rate of the patients.

Objective:Our study aims to predict acute kidney injury (AKI) in acute myocardial infarction (AMI) by establishing a random forest model.Methods:By using the clinical database from affiliated Dongyang Hospital of Wenzhou Medical University, a total of 1 363 AMI cases were included. Then, 75% of participants were analyzed as training subsets and the remaining 25% were testing subsets. The CARET package in R was used to filter variables and build random forest. The prediction ability of established model was evaluated by specificity, sensitivity, accuracy, relative operating characteristic curve (ROC curve) in testing subsets. In addition, the performance of random forest was compared with other 3 commonly used machine learning algorithms (Artificial Neural Network, Naive Bayes, and Support Vector Machine).Results:In this study, 30 variables including the demographic information, risk factors of cardiovascular disease, vital signs at admission, laboratory tests were identified and used to establish our random forest prediction model. The area under the curve of the testing subsets ROC was 0.893. The specificity and sensitivity of prediction model was 0.791 and 0.866, respectively. And the first creatinine, first blood urea nitrogen, and D-dimer after admission, age, mechanical ventilation were the top-five factors in this model. After comparing various machine learning algorithms, random forest model had a better performance.Conclusion:The random forest model would be used to predict the occurrence of AMI with AKI.

Objective To compare the safety and clinical efficacy of recombinant human tissue plasminogen activator (rt-PA) and urokinase(UK)in catheter-directed thrombolysis(CDT)for the treatment of subacute iliofemoral deep venous thrombosis(DVT). Methods From June 2013 to June 2017, a total of 116 subacute DVT patients underwent consistent CDT with either rt-PA or urokinase, or simple anticoagulation treatment in this study.The patients were divided into three groups for comparison:rt-PA-CDT group(n=43), UK-CDT group(n=39)and anticoagulation group(n=34). The baseline data, thrombolysis duration, rt-PA or UK dosages, thrombolytic rate and clinical efficacy rate were compared among the three groups. Independent t-test(accorded to normal distribution)was used to analyze the thrombolysis duration.The quantitative data were analyzed with analysis of varianc and the qualitative data were compared by the chi-square test. Results There was no significant difference in general clinical features among the three groups(P>0.05). The thrombolysis duration, total dosages and thrombolytic rate (≥50)were(5.8±1.3)d,(49.7±16.1)mg,86.0% for rt-PA-CDT group,and(6.3±1.5)d,(440±99)×104 U, 66.7% for UK-CDT group.The difference of thrombolysis duration was not statistically significant between the rt-PA-CDT group and UK-CDT group(t=-1.868, P>0.05). The thrombolysis rate of rt-PA-CDT group was significantly higher than that of UK-CDT group(χ2=4.315, P<0.05). The time of obtaining grade Ⅲthrombosis rate was shorter for rt-PA-CDT group(4.7±0.9)d compared with UK-CDT group(6.0±1.2 d) (t=-2.343,P<0.05).The clinical efficacy rates of the rt-PA-CDT group[88.4%(38/43)]and UK-CDT group [76.9%(30/39)]were significantly higher than that of anticoagulation group[26.5%(9/34)](P<0.05).There was no statistical difference between the rt-PA-CDT group and UK-CDT group(χ2=1.893, P>0.05). No severe complications were found in all groups. The incidence rates of mild complication of the rt-PA-CDT group, UK-CDT group and anticoagulation group were 16.3%(7/43), 17.9%(7/39)and 8.8%(3/34), respectively, and there were no significant differences among the three groups(χ2=1.396, P>0.05). Conclusion The clinical efficacy of CDT using rt-PA and UK for subacute DVT is better than simple anticoagulation treatment.Thrombolytic rate of rt-PA is superior to UK.

Objective To investigate the expression of CCL2 in colorectal cancer and its carcinogenesis mechanism associated with macrophages.Methods The expression level of CCL2 mRNA in 17 cases of colorectal cancer tissues and corresponding adjacent tissues were analyzed by PCR,and the phenotypes of macrophages in tumor infiltrating lymphocytes (TIL) were analyzed by flow cytometry.Human peripheral blood mononuclear cells were isolated and induced to differentiate into macrophages.After 12 h incubation with CCL2,the phenotypic changes of macrophages were analyzed by flow cytometry.The expression level of VEGF,COX-2 and IL-6 in the supernatant were measured by ELISA assay.Results The expression level of CCL2 in colorectal cancer tissues was significantly higher than that in corresponding adjacent mucosal tissues (t =4.017,P ＜ 0.05),and the macrophages in TIL had a high proportion of CCR2 phenotype.CCL2 was shown to induce increased CCR2 expression in macrophages (t =5.070,P ＜ 0.05),and promote the secretion of the tumor growth-associated factors such as VEGF,COX-2 and IL-6 (all P ＜ 0.05).Conclusion The levels of CCL2 in colorectal cancer were up-regulated suggesting that CCL2 may play a key role in tumor promotion by recruiting macrophages and influencing their function.

Due to the insufficient education of medical ethics and tending to medical moral educa-tion, the residents lack the ability to make ethical decisions. Therefore, we applied Four Topics method in the clinical ethics training of residents. The clinical ethics training was divided into three stages, and each stage had different training content and objectives. Dongyang People's Hospital adopted the form of lectures and panel discussions. During the group discussion, the group leader was in charge of case preparation, which came from clinical practice, and then all members applied the Four Topics method to analyze and discuss the ethical conflicts, and make the ethical decision. Through this process, residents' ability to solve ethical problems in clinical practice was enhanced.

Objective To discuss the clinical application of mechanical thrombectomy with AngioJet system for acute lower limb arterial ischemia (ALI).Methods A total of 12 ALI patients,who underwent percutaneous mechanical thrombectomy with AngioJet systemn during the period from January 2015 to November 2016,were enrolled in this study.The clinical data were retrospectively analyzed.The blood flow classification score after thrombolysis in myocardial infarction (TIMI) was used to evaluate the blood perfusion condition,and Cooley standard of efficacy score was used to assess the clinical curative effect.Results The technical success rate of mechanical thrombectomy with AngioJet system was 91.7％ (11/12).The average restored perfusion time was (1.5±0.6) hours.The clinical success rate and limb salvage rate were 83.3％ (10/ 12) and 91.7％ (11/12),respectively.The TIMI flow scores were improved from preoperative grade 0 (n=8) and Ⅰ (n=4) to postoperative grade 0 (n=1),Ⅰ (n=3) and Ⅱ (n=8).In 11 patients (91.7％) the symptoms of lower limb arterial ischemia were strikingly improved after mechanical thrombectomy.In one patient,the postoperative TIMI flow score remained 0 as preoperative state and the symptoms of lower limb arterial ischemia were not improved although catheter directed thrombolysis therapy was employed for 24 hours,and above-knee surgical amputation had to be carried out.Cooley efficacy score showed that complete cure was seen in 4 patients (33.3％),good response in 6 patients (50.0％),general improvement in one patient (8.3％) and pool response in one patient (8.3％).No severe bleeding complications occurred.Conclusion Percutaneous mechanical thrombectomy with AngioJet can rapidly recover the blood perfusion in patients with ALI,thus,further deterioration of the disease can be prevented and the limb salvage rate can be improved.Therefore,this technique has good clinical application value.

Objective Using chemically synthesized small interfering RNA (siRNA) transfected HepG2.2.15 cells to construct a cell model in interfering hepatitis B virus (HBV) X gene, studying the inhibi-tion of HBV replication and antigen expression in vitro. Methods After transfection of HepG2.2.15 cell for 24 h, 48 h, 72 h, detecting the cell supernatant of HBsAg and HBeAg by chemiluminescence immunoassay, the cell supernatant HBxAg protein by ELISA , the HBx mRNA relative expression of transfected cell was detected by fluorescence quantitative polymerase chain reaction (PCR), the ability of cell proliferation was detected by CCK-8 assay. Results After HBx-siRNA transfected HepG2.2.15 cells, cell proliferation ability was inhibited. The cell of HBx mRNA and the cell supernatant of HBxAg expression decreased (P < 0.05); at the same time it in-hibited the expression of HBsAg and HBeAg. The suppressed peak and the inhibition rate were 66% and 58%respectively at 72 h. The fluorescence quantitative PCR confirmed that expression of HBV DNA in the super-natant was decreased. Conclusion The HepG2.2.15 cell interference model of HBV X gene has been success-fully constructed, which has an effect of inhibiting proliferation of HepG2.2.15 cells and replication and expres-sion of HBV gene in vitro.

Objective An approach for analysis of hepatitis C virus (HCV) quasispecies using Hiseq high-throughput sequencing (hereinafter referred to as Hiseq sequencing) technique was developed and then applied to investigate a possible case of HCV needle sharing transmission. Methods One case of HCV antibody seroconversion (P1) was found in a methadone clinic on January 15, 2015. Four HCV antibody positive injecting drug users (IDUs), P2 to P5, suspected to be involved in needle sharing transmission with P1 during the period (after March 24, 2014) that P1 may be infected with HCV were investigated, and another 28 HCV antibody positive IDUs were selected as controls (C1 to C28). These controls came from the same methadone clinic or lived in the same town with P1. The RNAs were extracted from the plasma specimens and then reverse-transcribed into cDNA. After HCV subtyping, Hiseq sequencing was performed to detect and sequence the HCV quasispecies (263 bp) in the specimens with the same subtype as P1. The frequency of quasispecies was counted and ranked. Intrapersonal and interpersonal genetic distance and phylogenetic tree were calculated. Results The HCV subtype of specimen P1 was 3b. All the other specimens with the same subtype were P2, C7, C12, C14, C15, C16, C19, C20 and C28. Hiseq sequencing was successfully performed in 9 out of these 10 specimens, and 249 753 to 1 086 333 (average 869 608) cleaned sequences representing 3 to 172 (average 48) unique HCV quasispecies were obtained. The medians (P50) of intrapersonal genetic diversities from the 9 specimens were 0.4% to 12.3%. The P50 (P25, P75) of genetic diversities between P1 and the other 8 specimens were 19.0% (18.4%, 19.8%), 10.4%(2.8%, 18.3%), 19.6% (17.8%, 21.4%),24.9% (23.8%, 26.1%), 19.8% (18.7%, 20.7%), 20.1% (18.9%, 21.2%), 20.6% (20.0%, 21.1%), 23.6% (22.4%, 24.8%). There were no significant difference between the genetic diversities of P1 and P2 and those of P1 and other 7 specimens (H=9.40, P=0.100). The genetic diversities between few HCV quasispecies from P1 and few ones from C7 were 0. Phylogenetic tree analysis indicated that there was no HCV transmission relationship between P1 and P2, but there was HCV transmission relationship between P1 and C7. Conclusion With the feature of high-throughput, easier operation and lower cost, Hiseq sequencing technique has high practical value in tracing HCV transmission at the quasispecies level.

Objective An approach for analysis of hepatitis C virus (HCV) quasispecies using Hiseq high-throughput sequencing (hereinafter referred to as Hiseq sequencing) technique was developed and then applied to investigate a possible case of HCV needle sharing transmission. Methods One case of HCV antibody seroconversion (P1) was found in a methadone clinic on January 15, 2015. Four HCV antibody positive injecting drug users (IDUs), P2 to P5, suspected to be involved in needle sharing transmission with P1 during the period (after March 24, 2014) that P1 may be infected with HCV were investigated, and another 28 HCV antibody positive IDUs were selected as controls (C1 to C28). These controls came from the same methadone clinic or lived in the same town with P1. The RNAs were extracted from the plasma specimens and then reverse-transcribed into cDNA. After HCV subtyping, Hiseq sequencing was performed to detect and sequence the HCV quasispecies (263 bp) in the specimens with the same subtype as P1. The frequency of quasispecies was counted and ranked. Intrapersonal and interpersonal genetic distance and phylogenetic tree were calculated. Results The HCV subtype of specimen P1 was 3b. All the other specimens with the same subtype were P2, C7, C12, C14, C15, C16, C19, C20 and C28. Hiseq sequencing was successfully performed in 9 out of these 10 specimens, and 249 753 to 1 086 333 (average 869 608) cleaned sequences representing 3 to 172 (average 48) unique HCV quasispecies were obtained. The medians (P50) of intrapersonal genetic diversities from the 9 specimens were 0.4% to 12.3%. The P50 (P25, P75) of genetic diversities between P1 and the other 8 specimens were 19.0% (18.4%, 19.8%), 10.4%(2.8%, 18.3%), 19.6% (17.8%, 21.4%),24.9% (23.8%, 26.1%), 19.8% (18.7%, 20.7%), 20.1% (18.9%, 21.2%), 20.6% (20.0%, 21.1%), 23.6% (22.4%, 24.8%). There were no significant difference between the genetic diversities of P1 and P2 and those of P1 and other 7 specimens (H=9.40, P=0.100). The genetic diversities between few HCV quasispecies from P1 and few ones from C7 were 0. Phylogenetic tree analysis indicated that there was no HCV transmission relationship between P1 and P2, but there was HCV transmission relationship between P1 and C7. Conclusion With the feature of high-throughput, easier operation and lower cost, Hiseq sequencing technique has high practical value in tracing HCV transmission at the quasispecies level.

[ ABSTRACT] AIM:To investigate the effect of phycocyanin on the apoptosis of human laryngeal cancer HEP-2 cells and to explore the inhibitory mechanism of phycocyanin to tumor.METHODS:Highly purified phycocyanin was ex-tracted from spirulina.The effects of phycocyanin at different concentrations on the growth of human laryngeal cancer HEP-2 cells were detected by MTT assay.In addition, the cell structures were observed under electron microscope.The cell ap-optosis was analyzed by flow cytometry.The production of reactive oxygen species ( ROS) was measured by flow cytometry. Enzymatic activities of caspase-3,-8 and-9 were measured by chemical colorimatry.The expression of Bax, Bcl-2, Fas, P53, caspase-3 and caspase-9 at mRNA and protein levels was determined by RT-PCR and Western blot.RESULTS:MTT test confirmed that phycocyanin inhibited the cell activity of HEP-2 cells with time and dose dependent manners.The result of electron microscope observation and flow cytometry indicated that phycocyanin induced the apoptosis of HEP-2 cells.The intracellular content of ROS was increased.The activities of caspase-3, -8 and -9 were increased.RT-PCR showed that the mRNA expression of Bax, Fas, P53, caspase-3, caspase-9 was increased and Bcl-2 was decreased.The results of Western blot were consistent with the results of RT-PCR.CONCLUSION:Phycocyanin might induce apoptosis of HEP-2 cells by down-regulating Bcl-2, up-regulating Bax, Fas and P53, and the transduction of apoptotic signals in the human laryngeal cancer cells.