Lobaplatin induces apoptosis in T24 and 5637 bladder cancer cells by regulating Bcl-2 and Bax expression and inhibiting the PI3K/Akt signaling pathway.

Background: Lobaplatin (LBP) is a third-generation platinum-based drug that has been approved only in China for the treatment of several cancer types. Nonetheless, its efficacy in treating bladder cancer (BC) is unclear thus far. Through in vitro and in vivo experiments, this study aimed to explore whether LBP has an antitumor effect on T24 and 5637 BC cells and whether the effect is related to B-cell lymphoma-2 (Bcl-2), Bcl-2-associated X protein (Bax) and regulation of the phosphoinositide 3-kinase (PI3K)/protein kinase B (Akt) pathway. Methods: For in vitro experiments, the cell counting kit-8 (CCK-8) method was used to determine how different concentrations of LBP affect the viability of two types of BC cells. A wound healing assay was used to test the inhibitory effect of LBP on the migration of the two cell lines. Annexin V-fluorescein isothiocyanate isomer I (V-FITC)/propidium iodide (PI) staining was used to detect changes in cell apoptosis before and after LBP treatment, and Western blotting was used to detect the expression of apoptosis-related proteins and PI3K/Akt pathway proteins. For in vivo experiments, a cell-derived xenograft (CDX) model was employed, and the weight of nude mice and the tumor size were measured. Immunohistochemistry was used to detect the effect of LBP on the expression of apoptosis-related proteins in tumor xenografts. Results: In vitro, LBP reduced proliferation (P<0.05), inhibited migration (P<0.05), and induced apoptosis in T24 (31.25%±1.20%, P<0.01) and 5637 (14.3%±2.24%, P<0.05) BC cells, in a dose-dependent manner (P<0.05); increased the expression of proapoptotic proteins, including Bax, caspase-3 and cleaved caspase-3 (P<0.05); and suppressed the expression of antiapoptotic proteins, including Bcl-2, PI3K, Akt and phosphorylated Akt (p-Akt). The in vivo experiment confirmed that LBP can reduce the size of subcutaneous tumors in nude mice (P<0.05), increase the expression levels of Bax and cleaved caspase-3 and lower the expression of Bcl-2 (P<0.05) in bladder tumor tissue. Conclusions: The results obtained from both experiments suggest that LBP can inhibit the proliferation of T24 and 5637 BC cells, which might be credited to its effects in regulating Bcl-2 and Bax expression and inhibiting the PI3K/Akt pathway.

[The effects of anthocyanin on chronic inflammatory pain induced by complete Freund's adjuvant and its mechanism].

OBJECTIVE: To investigate the effects and mechanisms of anthocyanin from Ligustrum vicaryi on chronic inflammatory pain induced by complete Freund's adjuvant. METHODS: Thirty male SD rats were randomly divided into three groups (n=10):normal saline control group (NS), chronic inflammatory pain model group(Mod, injected with complete Freund's adjuvant(CFA) 100 µl to the left hind leg), anthocyanin treatment group(Ant, dosed with anthocyanins (90 m), mechanical pain threshold (MPT), and left toe volume in each group were measured before modeling and 1,3,5,7,9,11,13 days after operation. Antioxidant indexes in serum were mensurated by spectrophotometer, and the total capsaicin receptor (TRPV1) and phosphorylated capsaicin receptor (p-TRPV1) in hippocampus were detected by Western blot. RESULTS: In comparison with controls, HPT and MPT were improved (P<0.05),toe swelling was reduced(P<0.05), the serum level of SOD was increased (P<0.01), while the levels of MDA and NO were decreased (P<0.05), the ratio of P-TRPV1/TRPV1 protein was depressed in Mod rat hippocampal region treated with anthocyanin. CONCLUSIONS: The results show that anthocyanins has an analgesic effect on chronic inflammatory pain induced by CFA, and its mechanism may be related to the improvement of antioxidant capacity and the reduction of TRPV1 phosphorylation.

[Effect of bamboo leaf biochar addition on soil CO2 efflux and labile organic carbon pool in a Chinese chestnut plantation].

Effect of biochar addition on soil CO2 efflux in a typical Chinese chestnut (Castanea mollissima) plantation in Lin'an, Zhejiang Province, China was investigated from July 2012 to July 2013 by the static closed chamber-GC technique. Soil temperature, soil moisture, WSOC and MBC concentrations were determined as well. Results showed that soil CO2 efflux exhibited a strong sea- sonal pattern. Compared with the control (without biochar application), the biochar treatment increased the soil CO2 efflux only in the first month since application, and then the effect diminished thereafter. There were no significant differences in the annual cumulative value of soil CO2 efflux between the biochar and control treatments. The annual mean value in soil MBC concentration (362 mg · kg(-1)) in the biochar treatment was higher than that (322 mg · kg(-1)) in the control. However, no significant difference in the soil WSOC concentration was found between the biochar and control treatments. Strong exponential relationships between soil temperature and soil CO2 efflux were observed regardless of the treatment and soil layer. The apparent temperature sensitivity (Q10) of soil CO2 efflux in the biochar treatment was higher than that in the control. Soil CO2 efflux was related to soil WSOC concentration but not with soil MBC or moisture content. To conclude, the application of bamboo leaf biochar did not affect the annual cumulative CO2 emission in the Chinese chestnut plantation but increased the Q10, and the CO2 efflux was predominantly controlled by the soil temperature and soil WSOC level.

[Effects of fertilization on soil CO2 flux in Castanea mollissima stand].

In June 2011-June 2012, a fertilization experiment was conducted in a typical Castanea mollissima stand in Lin' an of Zhejiang Province, East China to study the effects of inorganic and organic fertilization on the soil CO2 flux and the relationships between the soil CO2 flux and environmental factors. Four treatments were installed, i. e., no fertilization (CK), inorganic fertilization (IF), organic fertilization (OF), half organic plus half inorganic fertilization (OIF). The soil CO2 emission rate was determined by the method of static closed chamber/GC technique, and the soil temperature, soil moisture content, and soil water-soluble organic carbon (WSOC) concentration were determined by routine methods. The soil CO2 emission exhibited a strong seasonal pattern, with the highest rate in July or August and the lowest rate in February. The annual accumulative soil CO2 emission in CK was 27.7 t CO2 x hm(-2) x a(-1), and that in treatments IF, OF, and OIF was 29.5%, 47.0%, and 50.7% higher than the CK, respectively. The soil WSOC concentration in treatment IF (105.1 mg kg(-1)) was significantly higher than that in CK (76.6 mg x kg(-1)), but was obviously lower than that in treatments OF (133.0 mg x kg(-1)) and OIF (121.2 mg x kg(-1)). The temperature sensitivity of respiration (Q10) in treatments CK, IF, OF, and OIF was 1.47, 1.75, 1.49, and 1.57, respectively. The soil CO2 emission rate had significant positive correlations with the soil temperature at the depth of 5 cm and the soil WSOC concentration, but no significant correlation with soil moisture content. The increase of the soil WSOC concentration caused by fertilization was probably one of the reasons for the increase of soil CO2 emission from the C. mollissima stand.