Objective To investigate the effects of chronic fluorosis on the expressions of matrix metalloproteinase-9 (MMP-9) mRNA and protein and the differentiation and maturation process of bone cell in the osteoclast of bone tissue of rats.Methods According to body weight,thirty-six healthy SD rats(body mass 100-120 g) were divided into three groups by random number table,twelve in each group,half male and half female.The rats of control group were given tap water(NaF ＜ 1 mg/L),and rats of low-fluorine and high-fluorine groups were fed with tap water containing 5 and 50 mg/L NaF to establish chronic fluorosis model.Rats were sacrificed after eight months; the contents of urinary fluoride in 24 hours and bone fluoride were analyzed by fluoride selective electrode.Serum content of tartrate resistant acid phosphatase 5b(TRACP5b)was detected by enzyme-linked immunosorbent assay (ELISA).The paraffin section of bone tissue was stained by hematoxylin-eosin (HE) and pathological morphometry was observed under optical microscope.The protein and mRNA levels of MMP-9 in the osteoclast of bones were detected by immunohistochemistry (IHC) and in situ hybridization (ISH),respectively.Results The differences of fluoride contents of urine and bone in rats were statistically significant between groups(F =400.612,48.229,all P ＜ 0.05).Fluoride contents of urine and bone were increased in lowfluorine and high-fluorine groups[(6.09 + 0.56),(7.69 + 0.64)mg/L,(12.65 ± 3.07),(26.53 + 5.88)mg/kg] compared to the control groups[(1.36 ± 0.51)mg/L,(0.67 ± 0.16)mg/kg,all P ＜ 0.05],and the fluoride contents of urine and bone were gradually increased with increasing fluoride doses(all P ＜ 0.05).The difference of TRACP5b content in serum was statistically significant between groups (F =9.607,P ＜ 0.05),in low-fluorine and high-fluorine groups,the TRACP5b contents[(1.86 ± 0.13),(1.92 ± 0.22)U/L] were higher than that of control group [(1.57 + 0.20)U/L,all P ＜ 0.05].The pathological examination showed osteosclerosis in fluoride exposed groups.The differences of MMP-9 mRNA and protein expressions were statistically significant between groups (F =365.727,331.382,all P ＜ 0.05).Compared to the control groups(97.22 ± 2.24,78.51 ± 1.16),the expressions of MMP-9 protein(108.18 ± 1.97,119.28 ± 1.76) and mRNA(89.44 ± 2.86,102.14 ± 2.39) were increased(all P ＜ 0.05),and the expressions of MMP-9 mRNA and protein were gradually increased with increasing fluoride doses (all P ＜ 0.05).Conclusions Chronic fluorosis might influence osteoclast differentiation and maturation process through regulating the expression levels of MMP-9 protein and mRNA.

Objective To investigate the effects of Shuangligan and Glycine on Thl/Th2 balancing on severe acute pancreatitis ( SAP) in rats. Methods Thirty-two Wistar rats weighing (260 ± 20) g were randomly divided into sham operation (SO) group, SAP group, SAP + Slg (with the treatment by Shuangligan) group and SAP + Gly (with the treatment by Glycine ) group. Each group included 8 rats, which accepted different treatment according to the experimental design. Changes of plasma level of endotoxin ( ET) and serum amylase (AMY) and the effects of Shuangligan and Glycine on Thl/Th2 ratio at the 24th hour after operation were observed respectively. Results The plasma endotoxin (ET) level ( (0. 67 ±0. 11) EU/ml),proinflammatory cytokine (INF-γ:(8.43 ± 0.86) ng/L, IL-12: (8.26 ± 1.97) ng/L) and Thl/Th2 ratio (0.36 ± 0.07) in SAP group were significantly higher than those in SO group( ET: (0. 44 ±0.07) EU/ml, INF-γ: (3. 80 ±0. 55) ng/L, IL-12: (3. 34 ± 1. 34)ng/L,Thl/Th2 ratio (0. 24 ±0. 05) ) (P <0. 05). Compared with SAP group, SAP + Slg and SAP + Gly group had remarkably decreased plasma ET level ( (0. 57 ± 0. 08,0. 52 ± 0. 04) EU/ml) (P < 0. 05) and the Thl/Th2 ratio reached equilibrium ( SAP + Slg group; (0. 29 ± 0. 04 ), SAP + Gly group: (0. 25 ± 0. 06 )) . Conclusions In the earlier stage of SAP, the rising plasma ET level may cause the overreaction of the cell mediated immune response, which leads to the aggravated damages in tissue cells. Our data indicates that Shuangligan and Glycine can restrain the formation of intestinal endotoxemia and alleviate or prevent the tissue injuries.

Multi-component Chinese medicine has been considered an important developmental direction for traditional Chinese medicine (TCM) formula.Nowadays,it is unfit to target a molecule for preventing or curing a disease because it resulted from multiple of factors and resulted in many indications.Thus,the idea focusing on the multiple targets for therapy is increasingly accepted by physicians and scientists.Here,we conceived a new simplified method for screening the active multi-component Chinese medicine based on the complexity of Chinese medicine,the multi-target property of protein signal transduction and the principle of Chinese medicine prescription.Combined with the traditional knowledge on tumor and latest antitumor research results of Chinese medicine and its compounds,the method was concretely illustrated.It helps us to transform the herbal compounds to new complex drugs targeting multiple signaling.

Objective To investigate the effect of intensive insulin therapy on long-term remittance of the islet β-cell function in newly diagnosed type 2 diabetic patients. Methods 120 newly diagnosed type 2 diabetic patients were randomly divided into four groups, and intensive insulin therapy was given for 15 days, 30 days, 60 days and 90 days respectively. The islet β-cell function were measured before and 1 or 2 years after treatment, and the differences were compared among each group. Results The plasma glucose was controlled well and the islet β-cell function was significantly improved in each group after treatment. The ratio value of △I30/△G30 in groups of 30 days,60 days and 90 days were higher than group of 15 days[(1.48±0.43 )mmol/L vs (1.25±0.40) mmol/L, t＝2.40,P＜0.05, (1.83±0.37) mmol/L vs (1.25±0.40) mmol/L, t＝2.85,P＜0.01, (1.90±0.41) mmol/L vs (1.25±0.40) mmol/L, t＝2.97,P＜0.01]. The indexes of the islet β-cell secretion function all gradually declined in each group after treatment for 2 years, but still higher than before treatment, the ratio value of △I30/△G30 in groups of 60 days and 90 days were higher than group of 15 days and 30 days[(1.44±0.51)mmol/L vs (0.87±0.47) mmol/L,t＝2.92, P＜0.01, (1.44±0.51)mmol/L vs (1.09±0.55) mmol/L, t＝2.44,P＜0.05, (1.52±0.44) mmol/L vs (0.87±0.47) mmol/L, t＝2.86, P＜0.01, (1.52±0.44) mmol/L vs (1.09±0.55) mmol/L, t＝2.50, P＜0.05], there was no difference between group of 60 days and 90 days. The ratio of remittance in groups of 60 days and 90 days was very high. Conclusions Intensive insulin therapy can significantly improve the islet β-cell function of newly diagnosed type 2 diabetic patients,anddelay the natural process. An appropriate extension of treatment can further prevent the descending rate of islet β-cell function, and easily get the long-term remission.

AIM: To demonstrate the effects of telmisartan on the proliferation and apoptosis of U937 cells.METHODS: The proliferation ability of the U937 cells was assessed by CCK-8 assay and colony formation test with methylcellulose.The CD11b expression rate of the U937 cells was identified by flow cytometry.The apoptotic rate was analyzed by flow cytometry with Annexin V-PI double staining and Hoechst 33342 staining.The protein levels of cleaved PARP and cleaved caspase-3 were determined by Western blot.RESULTS: The results of CCK-8 assay confirmed that the viability of U937 cells was inhibited by telmisartan.The colony formation capacity of U937 cells was also significantly inhibited by telmisartan.The differentiation of U937 cells was induced by telmisartan with the expression of CD11b.The results of flow cytometry analysis with Annexin V-PI double staining and Hoechst 33342 staining identified that the apoptosis of U937 cells was induced by telmisartan in dose-dependent and time-dependent manners with the up-regulation of cleaved PARP and cleaved caspase-3 proteins.CONCLUSION: Telmisartan inhibits the proliferation and induces the differentiation of U937 cells.Telmisartan also induces the apoptosis of U937 cells through the caspase pathway.

MTH1 (MutT Homolog1 )as MutT homologous en-zyme,is a nucleotide pyrophosphatase,mainly involved in DNA damage repair process,especially plays an important role in the process of DNA replication in tumor cells.Recent studies have found that MTH1’s function is responsible for the development of a variety of tumors.Studies have shown that,MTH1 can remove tumor cells’oxidative DNA elements detrimental to the function-al structure,protecting tumor cell division and proliferation, maintaining tumor cell survival,however,normal cells do not need MTH1.Therefore,MTH1 may be only closely associated with abnormal cell growth.This makes MTH1 as therapeutic tar-gets that have been paid much attention.This article reviewed the relationship of MTH1 and tumor,discussed the mechanisms of MTH1 in tumor growth MTH1 and tumor treatment,so as to provide reference for clinical research and treatment of tumor.

Objective To investigate the relationship between C-reactive protein (CRP) and carotid intima-media thickness in pa-tients with impaired glucose tolerance (IGT). Methods Senan CRP was measured with immunoturhidimetry and the carotid intima-media thickness (CAIMT) was measured using color Doppler in 108 patients with impaired glucose tolerance (IGT) and 80 subjects with normal glucose tolerance(NGT).Then we observed all IGT patients for 3 years using prospective follow-up method, Oral Glucose tolerance test (OGGT) and every index were measured in follow-up 1.5 year and 3 year. Results 2 objects were lost to follow-up. IGT group showed a significant higher CAIMT and CRP compared with NGT group. After follow-up 1.5 year and 3 year, the patients with impaired glucose toler-ance (IGT) were divided into type 2 diabetes (T2DM) group and IGT group based on the level of blood glucose. Both T2DM group and IGT group showed a significant higher CAIMT and CRP, compared with NGT group. The level of serum CRP of T2DM group was higher than that of IGT group, and the level of serum CRP of IGT group was higher than that of NGT group. There were great differences between each group.Linear correlation showed that the level of blood glucose was positively correlated with CRP and CAIMT in T2DM group after follow-up 3 year. CAIMT was positively correlated with the level of blood glucose and CRP. Mulfivariant stepwise regression showed that CRP was signifi-canfly correlated with the level of blood glucose and CAIMT. Conclusion Inflammation played an important role in the development of dia-betes, and it had great vessels complication. The patients with impaired glucose tolerance, who have high level of CRP, were facilitated to be diabetes, and they were at risk of getting great vessels complication during the phase of impaired glucose tolerance. So it would be helpful to prevent IGT patients with high CRP or CAIMT with anti-inflammatory therapy.

Aim To discuss the impact of important ac-tive ingredient of garlic diallyl sulfide———DATS on platelet activating factor (PAF ) mediated melanoma metastasis and its mechanisms.Methods ①MTT was used to test the effect of different concentrations of DATS on B16F10 and A375 melanoma cell growth number;②Scratch test and transwell were employed to test the effect of different concentrations of DATS on B16F10 and A375 melanoma cell migration;③ West-ern blot was used to test the effect of DATS on expres-sion of MMP-2,ERK,p38 induced by PFA;④Intrave-nous injection of tumor metastasis model was used to check the inhibition of DATS in PAF-mediated melano-ma metastasis.Results B16F10 cells relative growth rate fell to 73.21% and 48.78%,respectively,when DATS concentration reached 50 and 100 μmol·L-1 . DATS inhibited the levels of PAF-induced migration of melanoma cells B16F10 and vertical migration signifi-cantly,and inhibited B16F10 cells migration induced by PAF through inhibiting the expression of MMP-2, paxillin protein,FAK and other proteins.Conclusion DATS can significantly inhibit PAF-induced tumor metastasis, which is related to the inactivation of MAPKs.

Tumor metastasis is one of the most important biologi-cal characteristics of malignant tumor, and it is also the main factor resulting in poor prognosis and leading to failure of treat-ment. In recent years, studies have shown that the extracellular matrix ( ECM) of tumor microenvironment plays a critical role in tumor metastasis. Tumor ECM fibrogenesis could form the cross-linked network structure, which not only provides nutrition and support to tumor, also it is necessary to tumor growth and inva-sion. These research results indicate that blocking ECM fibro-genesis may exert an inhibitory effect on tumor metastasis. Therefore, targeting ECM fibrogenesis has become a particularly attractive strategy as it can be used in the treatment of metasta-sis-related diseases. The ECM fibrogenesis in tumor is reviewed in this paper as well as the treatment strategies on tumor metas-tasis by targeting ECM fibrogenesis, which may provide refer-ences for follow-up research and clinical treatment.

Objective To investigate whether TG2 promotes drug resistance to epirubicin through AKT signal pathway in breast cancer. Methods MCF-7 cells with constant expression of TGM2 gene(TGM2-LV)were established via the lentiviral vector. The breast cancer cells were divided into five groups,including the NC group, TG2 group and MK2206 group. The MCF-7/adr cells were divided into ADR group and MKadr group. The expres-sion of TG2 ,AKT ,Bcl-2 and P53 was detected by Western blot assay. Cells were treated with epirubicin. MTT assay was performed to assess cell proliferation. The inhibition ratio of cancer cell proliferation was evaluated. TUNEL analysis was performed to identify the apoptosis of the breast cancer cells. Results Lvels of TG2,p-AKT and Bcl-2 in NC group were significantly lower than those in TG2 group,while the expression of P53 in NC group was much higher. In MK2206(or MK/adr )group,p-AKT and Bcl-2 were down-regulated,while P53 was markedly up-regulated compared with TG2(or ADR)group(P<0.05). The results of the MTT assay showed a strong inhibi-tion in cell proliferation rate in MK2206(or MKadr )group. Compared with the NC group,TG2 promoted prolifera-tion of MCF-7 cells in TG2 group. The cell apoptosis rate in MK2206(or MKadr )group was significantly higher than that in TG2(or ADR)group(P<0.05). TG2 significantly inhibit the apoptosis of breast cancer cells ,com-pared to the control group. Conclusion TG2 might promote drug resistance to epirubicin through AKT signal path-way in breast cancer