Aim To investigate the effects of osthol on bone marrow stromal stem cells in vitro under the con-ditions of the ability to differentiate into osteoblasts and the case of proliferation. Methods The rat bone marrow sample was obtained,and the all bone marrow cell culture methods were used to separate and collect the stratum of mononuclear cells. The cells were cultured in DMEM containing 10% fetal bovine serum. Three days later the culture medium was changed for the first time. Nine days later,serial subcultivation proceeded. The final concentration of osthol was 1 ? 10 -4,1 ? 10 -5,1 ? 10 -6,1 ? 10 -7 mol?L -1 respectively. MTT method was adopted in proliferation analysis. Under the induced condition,the Alkaline phosphatase activity, calcium salt sediment yield and osteocalcin were meas-ured on 4th,8th,12th,16th day. On the fifteenth day,his-tochemistry dyeing proceeded for calcified tubercle. Total RNA was isolated and the gene expression of bFGF, IGF-1,Osterix and Runx-2 was investigated by RT-Real Time PCR. Results The BMSCs proliferation was refrained by osthol dose-dependently. But it evidently led to osteogenesis. The ALP activity,calcium salt sediment yield and osteocalcin were raised,and calcified tubercle amount was increased. Besides,it also could enhance the mRNA level of bFGF,IGF-1,Osterix and Runx-2. Conclusion The osthol with final concentra-tion of 1 ? 10 -5 mol?L -1 can markedly promote BM-SCs differentiation to osteogenesis. which proves osthol is an active constituent of the traditional Chinese medi-cine Common Cnidium Fruit.

Objective To investigate the combination effect of curcumin and γ-ray irradiation on PANC-1 cells in vitro.Methods PANC-1 cells were exposed to γ-rays in the presence or absence of curcumin.MTT assay was performed to evaluate cell viability.The expression of P21 was evaluated with RT-PCR and Western blot.Cell cycle distribution and apoptosis were tested by flow cytometry.Results Compared with the γ-ray irradiation group,combination treatment of curcumin and irradiation decreased the cell viability (t =6.72,P ＜ 0.01) and increased the percentage of cells in S-phase (t =4.78,P ＜ 0.05),apoptosis rate (t =6.58,P ＜ 0.01),P21 protein and mRNA expression (t =5.72,5.63,P ＜ 0.01) in PANC-1 cells.Conclusions Curcumin increases the radiosensitivity of PANC-1 cells,which may have clinical implication on radiotherapy of pancreatic cancer.

Aim To investigate the effects of K + channel blockers on arsenic trioxide-induced HeLa cell death. Methods Viability of HeLa cells was assessed by mitochondrial dehydrogenase activity using colorimetric MTT assay and the voltage-dependent K+ currents were recorded by using patch-clamp rest living cells after As2 O3 24 h-incubation showed significant increase of K + currents densities. At + 80mV, the densities of K+ currents (61 ± 18) pA/10 pF (n = 8) in As2O3 24 h-incubation group were significantly more than that in the control group (38 ± 10) pA/10 pF (n = 8, P ＜ 0. 05 ). The HeLa cells were prevented partially from As2 O3-induced cell death by co-application for 24 h with typical voltageeffects on HeLa cells. Conclusion Chronic treatment with As2 O3 increased voltage-dependent K+currents in HeLa cells and the cell death induced by As2O3 was reduced partially by voltage-dependent K +channel blockers, 4-aminopyridine or tetraethylammonium.

Objective To determine the toxicity, maximal dose and clinical practicality of VM26 (teniposide) plus radiotherapy for postoperative brain neurospongioma. Methods Twenty patients were alloted in phase Ⅰ trial. The total dose was 60 Gy for the field radiotherapy (30 fractions of 2 Gy over six weeks). Teniposide at three dose levels (50 mg/m2, 75 mg/m2 and 100 mg/m2) was given intravenously once a week, totally five weeks. Dose escalation was based on each level, with a minimum of five patients in cohort if severe toxicity was not observed until the maximum tolerance dose(MTD). Results The predominant form of toxicity was hematologic toxicity. Four patients developed grade 3, 4 leucopenia when a second level of MTD (75 mg/m2) was given. Conclusion Combined radiotherapy and teniposide for postoperative brain neurospongioma is well tolerated. The dose of 50 mg/m2 for phase Ⅱ clinical trial was recommended.

AIM: To investigate the effects of Xiaochaihutang (XCH), a Chinese medicine, and danazol on angiogenesis factor and microvessel density (MVD) of endometriosis (EM). METHODS: EM rats were treated with XCH, Danazol (D), and XCH plus D (group S) for 4 weeks, respectively. The histomorphology and volumes of ectopic endometrium were observed, the amount of macrophages in peritoneal fluid of EM model rats was counted, the concentration of interleukin-8 (IL-8) and tumor necrosis factor-? (TNF-?) of EM model rats in serum, peritoneal fluid, and supernate of cultured macrophages were measured. In addition, vascular endothelial growth factor (VEGF) was detected in ectopic endometrium by immunohistochemical SABC technique, MVD was determined by immunostaining for CD34. Similar studies were performed in rats without treatment (U group) and another group with sham operation (C). RESULTS: Compared with U group, XCH group, D group, and S group displayed a significant atrophy of ectopic endometrium, reduced number of endometrial glands, decreased macrophage amounts and low concentrations of IL-8 and TNF-? in blood, peritoneal fluid, and supernate of cultured macrophages . The expression of VEGF and MVD of ectopic endometrium in U group were significantly higher than that of C group. After treatment, they all decreased significantly, especially in S group. CONCLUSIONS: Both XCH and danazol can affect angiogenesis of EM model rats, cause an obvious atrophy in ectopic endometrium. XCH in combination with danazol can enhance the inhibitory effect on angiogenesis of EM model rats.

This study is to compare the effects of kaempferide and anhydroicaritin on biomineralization of rat osteoblasts (ROB) in vitro. Calvarias were dissected aseptically from newborn SD rats, the osteoblasts were obtained by enzyme digestion and were cultured in MEM containing 10% FBS. The medium was changed every three days, and serial subculture was performed when cells covered with 90% of the dish. Kaempferide and anhydroicaritin were separately added with final concentrations of 1 x 10(-4), 1 x 10(-5), 1 x 10(-6) and 1 x 10(-7) mol x L(-1) under the conditions of osteogenic differentiation. The proliferation was measured by MTT, and the optimal concentration was detected by the ALP activity at the 9th day after osteogenic induction culture. The osteogenic indexes of kaempferide, anhydroicaritin and control group with the optimal concentration were compared. The result showed that the anhydroicaritin at concentration of 1 x 10(-5) mol x L(-1) had significantly promoted the activity of ALP, calcium content and osteocalcin content, increased the number of CFU-F(ALP) and mineralized nodules, enhanced the mRNA level of BMP-2, OSX and Runx-2, which are key genes of osteogenic differentiation, and raised the protein content of collagen-I. However, the kaempferide group had not significantly represented the ability that promoted osteogenic differentiation of ROB. The difference of osteogenic differentiation on ROB between kaempferide and anhydroicaritin was caused by the prenyl group on C-8 of icariin.

Objective To quantitatively assess severity of movement disability by analyzing physical activities recorded by an actigraph monitor in patients with neurology disorders.Methods Eighty-one patients with Parkinson' s disease(PD)and 61 patients with acute cerebral infarction(ACI)accompanying impaired upper limb motor function were included in the study.PD patients and ACI patients were treated using the international PD and ACI treatment guidelines,respectively.The patients were asked to wear an Actigraph monitor for 6 days before the treatment in both PD and ACI patient groups and at 24-38 days post-treatment in PD patients or at 28 days post-treatment in ACI patients.The recorded data was analyzed by power-law exponent(PLE)and detrended fluctuation analysis(DFA).Clinically,before and after the treatments,PD patients were evaluated using the conventional Unified Parkinson Disease Rating Scale (UPDRS),and ACI patients were evaluated by assessing upper limb motor function using Fugl-Meyer Assessment(FMA)and Functional Independence Measure(FIM).The correlation of the UPDRS scores with PLE was analyzed in PD patients,and the correlation of FMA or FIM with DFA in ACI patients.Results Both the UPDRS scores and the PLE values in PD patients were improved after the drug administration(UPDRS total:32.8 ± 16.2 and 28.8 ± 14.7,Z =2.080,P =0.038; UPDRS Ⅲ:18.6 ± 8.2 and 15.7±6.8,Z=2.155,P=0.031; PLE:0.98 ±0.25 and 0.82 ±0.21,Z=2.212,P=0.027,before and after the treatment,respectively).There were a linear correlation coefficient of 0.699 between the improvements of total UPDRS scores and the PLE values,and of 0.823 between the UPDRS Ⅲ and the PLE values.FMA,FIM scores and DFA were improved significantly than before treatment(FMA:12.39 ± 8.21 and 30.28 ±7.29,Z=3.016,P =0.004; FIM:8.98 ±7.29 and 13.21 ±7.6,Z =2.282,P=0.038; DFA:0.86 ±0.31 and 0.98 ±0.27,Z =2.360,P =0.036,before and after the treatment,respectively).It also showed linear correlations between the improvements of FMA scores and DFA(r =0.638),and between FIM scores and DFA(r =0.712,both P ＜0.05).There was no correlation between UPDRS scores and DFA values in PD patients,nor between FIM scores or FMA scores and PLE values in ACI patients.Conclusions Actigraph device can be used to monitor patients activity in movement disorders.Analysis of its PLE can provide a quantitative evaluation in PD while its DFA may provide useful specific assessment of impaired upper limb motor function in ACI patients.It can also be used in quantitatively assessing new drug efficacy.

This study is to investigate the effects on the expression of iNOS and production of NO in the osteogenic differentiation process of rat bone marrow stromal cells (rBMSCs) by icariside II. rBMSCs were cultured by adherence screening method. When the culture dishes were covered with 80% cells, the osteogenic induced cultures were adopted. Icariside II was supplemented into the culture at 1 x 10(-5) mol x L(-1). The activity of iNOS, content of NO and osteogenic differentiation markers including alkaline phosphatase (ALP) activity, CFU-FALP and mineralized bone nodules were compared among the icariside II-supplemented group, L-NMAE group, icariside II + L-NAME group and the control. Total RNA was isolated and the gene expression of iNOS, Osterix and Runx-2 was investigated by real-time PCR. Total protein was also isolated and the secretion of iNOS and collagen I was examined by Western blotting. Icariside II can significantly improved ALP activity, CFU-FALP amount and mineralized nodules. Besides, the mRNA level of factors related to the osteogenic differentiation includes Osterix and Runx-2 also enhanced. The secretion of collagen I also promoted significantly. But all of these effects can be inhibited by L-NAME which can specifically inhibit the activity of iNOS. Icariside II enhances the osteogenic differentiation of rBMSCs significantly, but if the activity of iNOS was blocked by L-NAME, the osteogenic differentiation markers decrease accompanied with iNOS and NO decrease, suggesting that icariside II stimulates the osteogenic differentiation via enhancing the activity of iNOS and promoting the generation of NO.

AIM　To study the activation of choline on M3-R in heart and observe the hemodynamic changes of rat and rabbit. METHODS　A cardiac catheter was inserted into the left ventricular cavity via the right carotid artery, then the HR, LVSP, LVEDP, and ±dp/dt were measured using a polygraph system. RESULTS　Choline was shown to decrease the hemodynamic assessments, such as HR, +dp/dt, LVSP and LVEDP. while the M3-R antagonist 4-DAMP (4-diphenylacetoxy-N-methylpiperidine-methiodide) showed little effect on these assessments. It was found to reverse the hemodynamic effects of choline. CONCLUSION M3 receptor agonist can produce negative inotropic and chronotropic effects on the heart of rat and rabbit.

Aim To compare the pharmacological ac-tivity of icariin( ICA) and genistien ( GEN) against os-teoporosis after oral administration with them to growing rats and ovariectomized rats. Methods 25 mg·kg-1 icariin and 10 mg · kg-1 genistein ( equal in molar concentration) were administered to one-month-old fe-male SD rats every day for three months. Treatments at the same dosage were administered to the 6-month-old ovariectomized SD rats every day for three months. Their effects were compared on bone mineral density and biomechanical properties of femurs and vertebrae, serum levels of osteocalcin and tartaric acid phospha-tase 5b ( TRACP 5b) and histomorphometry. Results The results showed that, in young rats, icariin treat-ment significantly increased bone mineral density, the maximum mechanical loads of femurs and vertebrae as well as the bone qualities ( serum markers and microar-chitecture ) , whereas genistein treatment had little effects compared with the non-treatment control. How-ever, genistein treatment was more efficacious than icariin in preventing bone loss and deterioration of bone microarchitecture in ovariectomized rats. Conclusion Our data suggest that, since icariin has a higher os-teogenic activity but lower estrogenic activity, it has been found to be more efficacious than genistein in peak bone mass accrual only in young rats. In the ovariectimized rats, however, as the main force to pre-vent bone loss is the estrogenic activity, genistein has been found to be more efficacious than icariin in reduc-ing bone loss.