AIM:To further study the anti-tumor effect of angiostatin, an anti-human angiostatin monoclonal antibody was prepared and identified.METHODS:The hybrodoma techniques were used. The BALB/C mice were immunized with angiostatin. The supernatant of cell culture were collected and screened by ELISA and double immunodiffusion.RESULTS: There cell lines which steadily secreted the anti-angiostatin monoclonal antibody were identified by ELISA and double immunodiffusion. The antibody was IgG1 and specifically recognized angiostatin without crossing reactions to rhIL-2, rhTNF-?, rhIFN-? and serum proteins.CONCLUSION: The antibodies secreted by three hybridoma cell lines identified by several methods were specific antibodies of angiostatin.

Objective In this study,we aim to investigate and evaluate the application of peer education on the teaching of medical graduate students and to evaluate the teaching effect,in order to provide the basis for subsequent practice reform.Methods 49 graduate students majoring Internal Medicine-Pulmonology were randomly divided into traditional teaching(24) and peer education groups (25).We chose the primary culture technology of rat distal pulmonary arterial smooth muscle cells to be the teaching contents.For the traditional teaching group,we used the mode of class lecture giving and experimental skills learning under the assistance of the teachers; while in the peer education group,students benefited from the combination of class lecture given by the teacher and the seniors fixed teaching in which seniors help younger students.We selected the experimental operating time,cell purity and the practicing time to reach a standard culture as the evaluation indexes by filling a follow up questionnaire.The SPSS 13.0 was applied to the related data forx2 or t test.Results In the traditional teaching group,the average time to reach three times of standard culture was(3.2 ± 0.5) hour,which was(2.3 ± 0.4) hour in the peer education group.The cell purity was 80.1 ± 3.6％ in the traditional teaching group,while(85.4 ± 5.9)％ in the peer education group.The average practicing time was(6.3 ± 1.0) in the traditional teaching group,while(4.9 ± 0.6) in the peer education group.The peer education group master the teaching content better than the traditional teaching group (P=0.00).95.8％(23/24)of the students in the peer-education group considered the teaching contents simple,which was statistically higher(P=0.00) than traditional group (62.5％,15/24).Meanwhile,95.8％ (23/24)of the students in the peer-education group considered the teaching methods easy to accept,which was also statistically higher(P=0.02) than traditional group(70.8％,17/24).The difference was statistically significant (P=0.02).Conclusion The application effect of peer education is good and there is high degree of acceptance among the students.Besides,peer education accords with the medical postgraduate experiment teaching rules,and can cultivate medical graduate students' spirit of cooperation and communication ability in the process of implementation.

An aromatase encoded by the CYP19 gene catalyzes the final step in the biosynthesis of estrogens, which is related to endometriosis development. To assess the association of CYP19 gene polymorphisms with the risks of endometriosis, chocolate cysts and endometriosis-related infertility, a case-control study was conducted in Chinese Han women by recruiting 225 healthy control females, 146 patients with endometriosis, 94 endometriosis women with chocolate cyst and 65 women with infertility resulting from endometriosis, as diagnosed by both pathological and laparoscopic findings. Individual genotypes at rs2236722:T>C, rs700518:A>G, rs10046:T>C and [TTTA]n polymorphisms were identified. Allelic and genotypic frequencies were compared between the control group and case groups by chi-square analysis. Odds ratios (ORs) and 95% confidence intervals (95% CIs) were determined by logistic regression analysis to predict the association of CYP19 gene polymorphisms with the risk of endometriosis, the related chocolate cysts and infertility. The genotype distributions of the tested CYP19 gene polymorphisms were not significantly different between the healthy control group and the endometriosis/endometriosis with the chocolate cyst group. However, the CYP19 rs700518AA genotype was significantly associated with an increased risk of endometriosis-related infertility (55.4% in the infertility group vs 25.3% in the control group, P<0.001; OR (95% CI): 3.66 (2.06-6.50)) under the recessive form of the A allele. Therefore, we concluded that in Chinese Han females CYP19 gene polymorphisms are not associated with susceptibility to endometriosis or chocolate cysts, whereas CYP19 rs700518AA genotype confers genetic susceptibility to endometriosis-related infertility.
Adult ; Aromatase/*genetics ; Case-Control Studies ; China ; Endometriosis/complications/*genetics ; Female ; Genetic Predisposition to Disease ; Humans ; Infertility, Female/complications/*genetics ; Middle Aged ; *Polymorphism, Single Nucleotide

BACKGROUND AND OBJECTIVETransient receptor potential canonical (TRPC) proteins, a group of Ca2' permeable nonselective cation channels, are thought to constitute store-operated calcium channels (SOCC) and mediate store-operated calcium entry (SOCE) in various cell types. Members of TRPC have been found to be involved in abnormal proliferation, differentiation, and growth of cancer cells. The aim of this study is to detect the mRNA and protein expression of TRPC in non-small cell lung cancer (NSCLC).

METHODSReal-time quantitative PCRwas performed to screen the expression of TRPC mRNA in NSCLC tissue. Protein expression of TRPC was detected by Western blot.

RESULTSAmong the seven family members of TRPC so far identified (TRPC1-7), we detected the expression ofTRPC1, TRPC3, TRPC4, TRPC6 mRNA in 24 cases of NSCLC tissue; TRPC2, TRPC5 and TRPC7 mRNA were not detectable. The relative abundance of the expressed TRPC was TRPC1 approximately equal TRPC6 > TRPC3 > TRPC4. Western blot confirmed the protein expression of TRPC1, TRPC3, TRPC4 and TRPC6 in NSCLC tissue.

CONCLUSIONOut of the seven members of TRPC, we found TRPC1, TRPC3, TRPC4, TRPC6 mRNA and protein were selectively expressed in human NSCLC tissue. This study could provide a basis for future exploration of the individual role of these TRPC proteins in mediating SOCE and in the progression of lung cancer.

Adult ; Aged ; Blotting, Western ; Calcium ; metabolism ; Carcinoma, Non-Small-Cell Lung ; metabolism ; Female ; Humans ; Lung Neoplasms ; metabolism ; Male ; Middle Aged ; RNA, Messenger ; analysis ; TRPC Cation Channels ; genetics ; physiology

BACKGROUND: Lung cancer has a high incidence and mortality rate, but the treatment of lung cancer still lacks low toxicity and efficient anti-tumor drugs. Polysaccharide from radix tetrastigme has development value in anti-tumor treatment methods. This study was to observe the effect of polysaccharide from radix tetrastigme on immune response of Lewis lung cancer mice and explore its molecular mechanism. METHODS: Lewis lung cancer mouse models were established and randomly grouped. The spleen polypeptide group was intragastric with 50 mg/kg spleen polypeptide, and the radix tetrastigme polysaccharide low, medium and high dose groups were intragastric with 62.5, 125 and 250 mg/kg radix tetrastigme polysaccharide, respectively, and the model group and the control group were intragastric with equivolume normal saline. Tumor formation and metastasis were compared. Haematoxylin-eosin (HE) staining was used to observe the pathological changes of tumor cells. Macrophage phagocytosis, apoptosis, M1/M2 polarization, T cell subsets and cytokine levels in peripheral blood were detected by flow cytometry. The proliferation activity of macrophages was detected by methyl thiazolyldiphenyl tetrazolium (MTT) assay. Dendritic cell (DC) antigen presenting function was detected by chlorophenol red-β-D-galactopyranoside (CPRG) method. Tumor tissue differentiation antigen cluster 47 (CD47) mRNA and protein expression and macrophage signal regulatory protein α (SIRRP α) expression were detected by real time quantitative polymerase chain reaction (RT-qPCR) and Western blot (WB). RESULTS: The tumor inhibition rates and anti-metastasis rates in the 3-dose radix tetrastigme polysaccharide group and the spleen polypeptide group were higher than those in the model group, and the pathological injury of tumor tissue were severer, and the positive rate of phagocytosis of ink by macrophages and the efficiency of phagocytosis of tumor cells were increased; the apoptosis rate of macrophages was decreased; the proliferation activity of macrophages, polarization ratio of macrophages to M1 type, DC antigen presenting ability, CD4+, CD4+/CD8+ levels were increased; the level of serum tumor necrosis factor α (TNF-α), interleukin-1β (IL-1β), and the expression of tumor tissue CD47, macrophage SH2-containing protein tyrosine phosphatase 1 (SHP-1), SH2-containing protein tyrosine phosphatase 2 (SHP-2), and phosphorylation signal regulatory protein α (p-SIRPα) were decreased, and the differences were statistically significant (P<0.05). There were no significant differences in the above indexes between low-dose radix tetrastigme polysaccharide group and spleen polypeptide group (P>0.05), and the effects of radix tetrastigme polysaccharide were dose-dependent. CONCLUSIONS Radix tetrastigme polysaccharide can inhibit tumor growth, metastasis and immune response in Lewis lung cancer mice, and its mechanism may be related to inhibiting SIRP/CD47 signaling pathway.
Mice ; Animals ; CD47 Antigen/genetics* ; Lung Neoplasms/drug therapy* ; Cytokines/genetics* ; Polysaccharides/pharmacology* ; Immunity ; Protein Tyrosine Phosphatases

Objective: To establish an animal model of hypoxia-induced bronchopulmonary dysplasia asso-ciated with pulmonary hypertension (BPD-PH). Methods: C57BL/6 male and female specific pathogen free mice mated and female mice with their offspring mice were randomly divided into normoxic group and hypoxia group by way of numerical method.Normoxic group was placed in the indoor environment directly.Hypoxia group was placed in 120 mL/L oxygen concentration environment within 12 hours after birth.Body weight gain and mortality of the neonatal mice were recorded.The mice lungs and hearts were harvested on day 14 for immunofluorescence staining and HE staining, and Western blot was used to observe the morphological changes and vascular endothelial growth factor (VEGF) protein level. Results: The mortality rates of normoxic group and hypoxic group were 11.8% and 47.3%, respectively.Compared with the normoxic group, body weight of hypoxia group increased slowly, as the final body weight of 2 groups were (12.40±2.33) g and (5.50±0.32) g, respectively, and the difference was significant (χ²=13.38, t=20.50, all P<0.01). Hypoxia group showed higher right ventricular hypertrophy index [(96.00±0.15)% vs.(40.40±4.00)%, t=41.67, P<0.01], fusion of alveolar, diminished microvasculature, thickening alveolar septal, decreased alveolar radiation coefficient (RAC)(19.73±2.33 vs.10.90±1.85) and increased mean linear intercept(MLI) (33.2±4.33 vs.58.70±7.27) with statistical significance, respectively (t=16.27, 9.53, all P<0.01). In the lung HE staining, pulmonary arterial thickening, pulmonary smooth muscle cell proliferation and intimal roughness in immunofluorescence, significantly decreased expression of VEGF protein level by Western blot were observed in hypoxic group (0.41±0.04 vs.1.19±0.08, t=15.10, P<0.01). Conclusions Hypoxia-induced mouse BPD-PH model was consistent with the pathophysiological process of pulmonary vascular remodeling of pulmonary hypertension, which increase pulmonary vascular resistance eventually leading to right ventricular hypertrophy.