Objective To identify-and study a monoclonal antibody (McAb) against pancreatic cancer stem cell in vitro,as well as to provide candidate antibody-drug for cancer stem cell-targeted therapy of pancreatic cancer.Methods Cell culture in serum-free medium and PKH26 staining were used to determine the existence of cancer stem cell in PANC-1 cell line.Flow cytometry was used to detect the expression of CD24 and CD44 in PANC-1 cells and sphere cells,Immunofluorescence was used to detect the expression of CD24 and antigen recognized by 15D2.The effects of 15D2 on self-renewal,proliferation and chemosensitivity to gemcitabine of PANC-1 parent or sphere cells were identified by serum-free suspension culture and CCK-8 assay,Immunohistochemistry was applied to detect the level of antigen recognized by 15D2 in cancer and adjacent tissues.Results PANC-1 cells could survive,proliferate and form sphere cells in serum-free medium.The sphere-forming rate was (2.5±0.5) ％.The percentage of CD44+ CD24+ cells population in sphere cells increased by 11.4 folds compared to PANC-1 cells,in which single nearly 97 ％ CD24+ cells was CD44+ CD24+ cells.Therefore,CD24+ was selected for cancer stem cell marker in PANC-1 in this study.The two-color immunofluorescence assay showed that 15D2 could recognize cells which was also stained by CD24.In vitro functional experiments demonstrated that 15D2 significantly suppressed the sphere formation of PANC-1 cells,with the inhibitory rate being 30.4 ％.Meanwhile,the combination of 15D2 and gemcitabine can significantly attenuate the growth of PANC-1 sphere cells.The IC50 was 0.10 μmol/L in 15D2+gemeitabine group,and 0.39 μmol/L in mlgM+gemcitabine group,Immunohistochemical results showed that the antigen recognized by 15D2 was greatly expressed in about 76.9 ％ (11/13) human pancreatic cancer tissues and hardly detected in adjacent normal tissues (10.0 ％,1/10).Conclusion McAb 15D2 can inhibit self-renewal and drug-resistance of pancreatic cancer stem cell in PANC-1 cell line,and it might become a candidate drug for target pancreatic cancer stem cell treatment.

Objective　To evaluate the expression of MAGE-1 gene in esophageal carcinoma and determine whether esophageal carcinoma patients should be eligible for MAGE-1 antigen-based vaccine therapies． Methods　 MAGE-1 mRNA expression in esophageal carcinoma was assessed by reverse transcription and polymerase chain reaction amplification． The PCR products were then digested by restriction endonucleases and inserted into the pET-30a(+) vector． The sequence of the inserted gene fragment was confirmed by DNA sequence analysis． Results　Out of the 30 esophageal carcinomas studied, 43% of the esophageal carcinomas tissues expressed MAGE-1 gene and no expression was found in matched adjacent normal esophageal mucosae． The entire cDNA of the gene was cloned． Conclusion　Owing to the high frequency of MAGE-1 gene expression in esophageal carcinoma and MAGE-1 antigen can be recognized by cytolytic T lymphocytes when presented by class-I HLA molecular, a large proportion of these patients might be suitable candidates for immune therapy involving tumor specific antigens encoded by MAGE-1 gene．

OBJECTIVE:To systematically review the efficacy and safety of cinacalcet in the treatment of hemodialysis pa-tients with secondary hyperparathyroidism,and provide evidence-based reference for the clinical treatment. METHODS:Retrieved from Medline,Cochrane Library,EMBase and CBM,randomized controlled trials(RCT)about cinacalcet in the treatment of he-modialysis patients with secondary hyperparathyroidism (SHPT) were collected. Meta-analysis was performed by using Rev Man 5.3.5 software after data extract and quality evaluation by Cochrane systematic Rev Man 5.3.5. RESULTS:Totally 7 RCTs were en-rolled,involving 1 987 patients. Results of Meta-analysis showed cinacalcet can significantly reduce the rate of surgical parathyroid-ectomy[RR=0.23,95%CI(0.06,0.89),P=0.03],incidence of fracture[RR=0.26,95%CI(0.12,0.60),P=0.002] and increase the incidences of hypocalcemia[RR=9.81,95%CI(3.92,4.59),P<0.001],nausea[RR=1.97,95%CI(1.58,2.46),P<0.001] and vomit-ing[RR=1.91,95%CI(1.50,2.42),P<0.001],while it showed no significant effect on the the incidence of all-cause mortality and cardiovascular death. CONCLUSIONS:The clinical efficacy of cinacalcet in the treatment of hemodialysis patients with secondary hyperparathyroidism is good,but there are common adverse reactions such as nausea and vomiting,hypocalcemia.

OBJECTIVETo investigate the the expression and hypoxic regulation of vascular endothelial growth factor(VEGF) and matrix metalloproteinase-9.

METHODSVEGF mRNA and MMP-9 mRNA were examined by reverse transcription-polymerase chain reaction (RT-PCR) in 43 esophageal carcinoma specimens including 18 para-tumorous esophageal tissues. The expression of VEGF protein and mean microvessel density (MVD) in 56 specimens were examined by immunohistochemical stain. The effect of hypoxia on VEGF and MMP-9 expression in esophageal cancer cell lines was quantitatively determined by enzyme linked immunosorbent assay (ELISA).

RESULTSThe VEGF expression in the tumorous tissue, being significantly correlated with MVD in the tumor, was remarkably higher than that in the para-tumorous tissue. VEGF and MVD expression in the tumor was significantly associated with stage and metastasis of esophageal carcinoma. The MMP-9 expression in the tumorous tissue, being uncorrelated with vessel count and clinicopathologic features in esophageal carcinoma, was significantly higher than that in the para-tumorous tissue. Hypoxia significantly increased the VEGF expression in esophageal cancer cell lines but did not affect the MMP-9 expression.

CONCLUSIONSThe expression of VEGF plays an important role in the angiogenesis and metastasis of esophageal cancer, which is regulated by hypoxia. VEGF may serve as a predictor of progression in esophageal carcinoma and a potential target for antiangiogenic therapy of esophageal carcinoma.

Endothelial Growth Factors ; biosynthesis ; genetics ; Esophageal Neoplasms ; metabolism ; Female ; Gene Expression Regulation ; Humans ; Hypoxia ; Lymphokines ; biosynthesis ; genetics ; Male ; Matrix Metalloproteinase 9 ; biosynthesis ; genetics ; Middle Aged ; Oxygen ; metabolism ; RNA, Messenger ; biosynthesis ; Reverse Transcriptase Polymerase Chain Reaction ; Vascular Endothelial Growth Factor A ; Vascular Endothelial Growth Factors

Objective: To analyze the pulmonary valve function in patients with tetralogy of Fallot after radical surgery. Methods: Clinical data of 263 patients (119 male, mean age (33.2±11.5) years old) with tetralogy of Fallot underwent radical surgery in our hospital from January 2010 to June 2016 were retrospectively analyzed. According to age, patients were divided into 14-17 years old group (14 cases), 18-29 years old group (100 cases), 30-39 years old group (61 cases) and above 40 years old group (87 cases). The patients were divided into pulmonary regurgitation group (87 cases) and control group (176 cases) according to weather they have moderate or severe pulmonary regurgitation. Echocardiographic data were compared among groups. Results: A total of 83 patients received re-operation. The median age of the primary radical operation was 9 (5, 13) years, and the median time from the primary radical operation to echocardiographic follow-up was 5 (1, 13) years. Among the 263 enrolled patients, prevalence of pulmonary regurgitation was 36.1% (95/263), and pulmonary stenosis was evidenced in 28 patients (10.6%). The ratio of moderate to severe tricuspid regurgitation was 14.3% (2/14), 27.0% (27/100), 32.8% (20/61) and 37.9% (33/87) in 14-17 years old group, 18-29 years old group, 30-39 years old group and above 40 years old group, respectively (P=0.029), while prevalence of moderate and severe pulmonary regurgitation, moderate and severe pulmonary valve stenosis, pulmonary valve transvalvular pressure >40 mmHg (1 mmHg=0.133 kPa), right atrial and right ventricular enlargement ratio were similar among groups (all P>0.05). The ratio of moderate and severe tricuspid regurgitation and right ventricular enlargement in the pulmonary regurgitation group was significantly higher than in the control group (40.2% (35/87) vs. 27.3% (48/176) and 96.6% (84/87) vs. 87.5% (154/176), all P<0.05), while left ventricular ejection fraction, right atrial enlargement, and right ventricular wall thickness were similar between the two groups (all P>0.05). Conclusion Pulmonary regurgitation is a common clinical feature among survivors of tetralogy of Fallot patients after radical surgery, and moderate to severe pulmonary regurgitation increases the risk of tricuspid regurgitation and enlargement of the right ventricle.

Objective To explore the risk factors and nursing intervention for pneumothorax in performing CT-guided radioactive 125I seed implantation in patients with lung cancer.Methods 40 patients with lung cancer who performed CT-guided radioactive 125I seedimplantation from January 2014 to November 2015 were included in the study. The retrospective analysis was used and the participants were hospitalized either in Peking university third hospital or in Hebei Cangzhou hospital of integrated traditional Chinese and western medicine. Results After the implantation, seven patients had implantation including five mild cases and two moderate cases. Six patients got better after the implantation (i.e. five of them had no obvious clinical symptoms and one of them had chest distress and cough). One case had hemothorax combined with chest distress and cough, but got better after air extraction and closed thoracic drainage.Conclusions The risk factors for pneumothorax in performing CT-guided radioactive 125I seed implantation may include the type and location of the lesion and the number of puncture times. The important factors for preventing the incidence of pneumothorax may include preoperative analysis of the type and location of lesions, reducing thepuncture times, less harm for pleural layers,and reasonable needlepoint. The major nursing strategies to prevent pneumothorax include preoperative evaluation and health education, cooperate with the doctors during the operation,and close observation of patients'postoperative symptoms.

Objective To investigate the biological characteristics of monoclonal antibodies against human liver cancer stem cells and its therapeutic effect in combination with cisplatin in the treatment of hepatocellular carcinoma. Methods Cell culture in serum?free medium and PKH26 staining were used to determine the existence of cancer stem cells in human liver Bel7402?V3 cell line. The co?expression of antigen recognized by monoclonal antibody ( McAb ) 15D2 and epithelial specific antigen ( ESA ) and PKH26?positive cells in the Bel7402?V3 cells were detected by immunofluorescence assay. Serum?free suspension culture was used to detect the self?renewal ability of 15D2?positive Bel7402?V3 cells sorted by flow cytometry and the effect of 15D2 on the self?renewal ability of Bel7402?V3 cells. The effect of 15D2 on cisplatin resistance in the cells was examined by CCK8 method. The inhibitory effect of 15D2 combined with cisplatin on the transplanted tumor growth in mice was also observed. Results Single PKH26?positive cells were observed in the Bel7402?V3 cell spheroids cultured for 11 days. Immunofluorescence assay showed that the 15D2?recognized antigen could be conjugated with PKH26 and ESA and co?localized on Bel7402?V3 cells. The spheroid formation rate of 15D2?positive cells in serum?free medium was significantly higher than that of 15D2?negative cells [(30.4±3.4)% vs. (8.8±1.8)%,P<0.01]. The cisplatin resistance of 15D2?positive cells was obviously higher than that of 15D2?negative cells (IC50:1.014μmol/L vs. 0.365μmol/L). McAb 15D2 significantly suppressed the spheroid formation of Bel7402?V3 cells, with an inhibition rate of 37.5%. McAb 15D2 also notably inhibited the cisplatin resistance of Bel7302?V3 cells. The IC50 was 0.211μg/ml in the 15D2 group and 0. 325 μg/ml in the control group. The mouse experiment showed that the tumor growth rates of 50 mg/kg, 25 mg/kg and 12.5 mg/kg 15D2?treatment groups were 82.6%, 71.4% and 60.0%, respectively; that of the 50 mg/kg 15D2 + cisplatin group was 91. 0%, and that of the cisplatin monotherapy was 56. 7%. Conclusion McAb 15D2 is a functional monoclonal antibody targeting liver cancer stem cells, which could be a potential monoclonal antibody drug for the stem cell?targeted therapy of liver cancer.

Objective To investigate the biological characteristics of monoclonal antibodies against human liver cancer stem cells and its therapeutic effect in combination with cisplatin in the treatment of hepatocellular carcinoma. Methods Cell culture in serum?free medium and PKH26 staining were used to determine the existence of cancer stem cells in human liver Bel7402?V3 cell line. The co?expression of antigen recognized by monoclonal antibody ( McAb ) 15D2 and epithelial specific antigen ( ESA ) and PKH26?positive cells in the Bel7402?V3 cells were detected by immunofluorescence assay. Serum?free suspension culture was used to detect the self?renewal ability of 15D2?positive Bel7402?V3 cells sorted by flow cytometry and the effect of 15D2 on the self?renewal ability of Bel7402?V3 cells. The effect of 15D2 on cisplatin resistance in the cells was examined by CCK8 method. The inhibitory effect of 15D2 combined with cisplatin on the transplanted tumor growth in mice was also observed. Results Single PKH26?positive cells were observed in the Bel7402?V3 cell spheroids cultured for 11 days. Immunofluorescence assay showed that the 15D2?recognized antigen could be conjugated with PKH26 and ESA and co?localized on Bel7402?V3 cells. The spheroid formation rate of 15D2?positive cells in serum?free medium was significantly higher than that of 15D2?negative cells [(30.4±3.4)% vs. (8.8±1.8)%,P<0.01]. The cisplatin resistance of 15D2?positive cells was obviously higher than that of 15D2?negative cells (IC50:1.014μmol/L vs. 0.365μmol/L). McAb 15D2 significantly suppressed the spheroid formation of Bel7402?V3 cells, with an inhibition rate of 37.5%. McAb 15D2 also notably inhibited the cisplatin resistance of Bel7302?V3 cells. The IC50 was 0.211μg/ml in the 15D2 group and 0. 325 μg/ml in the control group. The mouse experiment showed that the tumor growth rates of 50 mg/kg, 25 mg/kg and 12.5 mg/kg 15D2?treatment groups were 82.6%, 71.4% and 60.0%, respectively; that of the 50 mg/kg 15D2 + cisplatin group was 91. 0%, and that of the cisplatin monotherapy was 56. 7%. Conclusion McAb 15D2 is a functional monoclonal antibody targeting liver cancer stem cells, which could be a potential monoclonal antibody drug for the stem cell?targeted therapy of liver cancer.

In 2020, due to the impact of the novel coronavirus epidemic, the development of transcatheter heart valve therapy has been shown to slow down, but there are still many aspects worth noting. The indication of monoclonal antibody after transcatheter aortic valve replacement (TAVR) should be further clarified. Low surgical risk patients were included in TAVR relative indications. Mitraclip G4 was approved by CE. The indication of atrial septal occlusion after mitraclip should be further clarified. The technique of coaptation augmentation is expected to become a new method of mitral valve interventional repair. Tendyne transcatheter mitral valve was approved by European Union. Transcatheter tricuspid valve treatment equipments, TriClip and PASCAL obtained CE mark. TAVR technology is being popularized rapidly in China, and what’s more, balloon dilated valve Sapien 3 and new recyclable repositioning valve system-Venus plus have entered the domestic market. A number of mitral valve therapeutic instruments have appeared one after another, and China's first tricuspid valve lux has completed its FIM research. Finally, with the improvement of devices and technology in the future, interventional therapy of heart valve is expected to benefit more patients.

Objectives:To observe the mitochondrial morphology of normal and triple-negative breast cancer cells, extract mitochondria from normal cells, and investigate the effects of mitochondrial transplantation on proliferation, apoptosis, and stemness of triple-negative breast cancer cells.Methods:The morphology of mitochondria was observed by transmission electron microscope. Mitochondria were extracted by mitochondrial extraction kit, mitochondrial protein was identified by western blot, and mitochondrial activity was detected by mitochondrial membrane potential detection kit. MitoTracker Green or MitoTracker Deep Red fluorescent probes were used to label the mitochondria of living cells, and the degree of mitochondria entering LTT cells was observed by confocal laser microscopy at 12, 24, and 96 hours. The effects of mitochondrial transplantation on proliferation, apoptosis, and stemness of breast cancer cells were examined by CCK8, colony formation assay, flow cytometry, and sphere formation assay after 24 hours of mitochondrial transplantation.Results:The mitochondria of normal cells were rod-shaped or elongated, while the mitochondria of triple-negative breast cancer cells were swollen and vacuolated. Western blot results showed that cytochrome c oxidase subunit I (MT-CO1) protein encoded by mitochondria was present in the isolated mitochondria. The content of heat shock protein 60 (HSP60) was higher in mitochondria than that in cytoplasm. The result of the multi-mode microplate reader showed that the content of mitochondrial J-aggregates/monomer was 1.67±0.06, which was significantly higher than 0.35±0.04 of the control group ( P＜0.001). Exogenous mitochondria were observed in LTT cells at 12, 24, and 96 hours after mitochondrial transplantation. The results of the CCK8 experiment showed that OD450 of LTT cells was 0.27±0.13 after 48 hours transplantation, which was lower than 0.62±0.36 of the control group ( P=0.023). The OD450 of MDA-MB-468 cells was 0.30±0.03, which was lower than 0.65±0.10 of the control group ( P=0.004). After 120 hours of mitochondrial transplantation, OD450 in both groups was still significantly lower than that in the control group (P＜0.01). The number of clones formed by mitochondrial transplantation of LTT cells was 21.33±7.31, which was lower than 35.22±13.59 of the control group ( P=0.016). Flow cytometry showed that the early apoptosis rate of LTT cells was (30.07±2.15)% after 24 hours of mitochondrial transplantation, which was higher than 2.07±1.58 of the control group ( P＜0.001). The proportion of early apoptosis in MDA-MB-468 cells was 24.47%±5.22%, which was higher than (7.83±2.06)% in the control group ( P=0.007). In addition, the number of mitochondria transplanted LTT cells into the cell sphere was 46.25±5.40, which was significantly lower than 62.58±6.43 of the control group ( P＜0.001). Conclusion:Normal mitochondria can enter triple-negative breast cancer cells by co-culture, inhibit the proliferation and stemness of triple-negative breast cancer cells, and promote the apoptosis of triple-negative breast cancer cells.