Objectives To investigate the effects of matrine on the proliferation and apoptosis of SK-NEP-1 cells in vitro, and its possible mechanism. Methods Trials were divided into following groups:control group, 0.5, 1.0 and 1.5mg/ml of ma-trine intervention groups. The inhibition rate of SK-NEP-1 cells treated with different concentration of Matrine was detected by MTT colorimetric assay. Apoptosis rate was detected by flow cytometry (FCM). RT-PCR analysis was employed to measure the PDCD4 mRNA expression. Results Matrine (final concentrations=0.5, 1.0 and 1.5mg/ml) could induce apoptosis and inhibit the growth of SK-NEP-1 cells. Compared with the controls without matrine treatment (8.81±3.71)%, the inhibition rates of SK-NEP-1 cells were (20.79 ± 6.20)%, (31.25 ± 5.07)%, and (51.15 ± 12.70)%, respectively;the apoptotic rates of SK-NEP-1 cells treated with different concentration of matrine were (13.67±0.78)%,(17.43±1.65)%and (20.80±1.54)%, respectively. Significant difference in the inhibition and apoptotic rates of SK-NEP-1 cells between each drug group and control group was observed(P<0.05), and the inhibition and apoptotic rates of SK-NEP-1 cells increased gradually with increased matrine concentration, thus exhibiting a dose-dependent effect(P<0.05). To the expression of CXCR4 mRNA,the grey levels of SK-NEP-1 cells treated with matrine intervention group (final concentrations=0.5, 1.0 and 1.5 mg/ml) were (0.720 ± 0.058), (0.540 ± 0.095) and (0.307 ± 0.050), respectively. The mRNA expression of CXCR4 was seen in SK-NEP-1 cells. Compared with control group, the expres-sion of CXCR4 mRNA was decreased significantly in matrine intervention group (P<0.01).There were significant difference in CXCR4 mRNA level among the SK-NEP-1 cells treated with 0.5,1.0,1.5mg/mL of matrine (P<0.01). Conclusions Matrine could induce apoptosis and inhibit the growth of SK-NEP-1 cells in a dose-dependent way which may be associated with the down-regulated CXCR4 expression in SK-NEP-1 cells.

Objective To investigate the effects of matrine on proliferation and apoptosis of human rhabdomyosarcoma(RMS) RD cells line in vitro,to study the regulatory mechanism of Wnt/β-catenin pathway-influenced apoptosis of RD cells line by detecting the expressions of β-catenin protein,Bcl-2 protein and caspase-3 protein,and to explore Wnt/β-catenin mechanism during the process of RMS.Methods The human RMS RD cells line was treated with matrine of different concentrations (0.5,1.0,1.5,2.0 g/L)for 48 hours respectively,and the proliferation inhibition rates of different concentrations of matrine on RD cells were detected by methyl thiazolyl tetrazolium assay,while the apoptosis rates by flow cytometry (FCM) and the expressions of β-catenin,Bcl-2 and caspase-3 by Western blot.Results The proliferation inhibition rates between control group and different concentrations of matrine groups were(13.70 ±0.25)％,(33.16 ±0.11)％,(42.96 ±0.90)％,(56.26 ±0.79)％ and (67.89 ±0.63)％,respectively.The apoptosis rates were (5.49 ± 0.96) ％,(17.23 ± 5.03) ％,(25.84 ± 4.17) ％,(36.08 ± 3.68) ％and (47.79 ± 4.82) ％,respectively.The highest expression of β-catenin and Bcl-2 proteins and the minimum amount of caspase-3 protein were found in the control group.After intervention of matrine,the expressions of β-catenin and Bcl-2 reduced while the amount of caspase-3 rose significantly,which was concentration-dependent obviously.Differences were found between every concentration of matrine group with control group according to statistics (all P ＜ 0.05).Conclusions Matrine can inhibit proliferation and induce apoptosis of RD cells.Matrine can down-regulate the expression of β-catenin and Bcl-2 proteins in RD cells,while the amount of caspase-3 protein rises.Wnt/β-catenin signal pathway plays an important role in the apoptosis of RD cell induced by matrine,and its downstream proteins Bcl-2 and caspase-3 are also involved in the regulation of this process.

Objective To investigate the effect of Matrine on the expression of X-linked inhibitor of apoptosis protein (XIAP) in human rhabdomyosarcoma RD cells in vitro.Methods Cultured human rbabdomyosarcoma RD cells were divided into Matrine intervention groups (0.5 g/L,1.0 g/L and 1.5 g/L) and a control group.The proliferation-inhibition rates in RD cells treated with different concentrations of matrine were detected by methylthiazolyl blue colorimetric assay.Flow cytometry analysis was performed for the apoptosis rates of RD cells.Reverse transcription-polymerase chain reaction analysis was used to measure the XIAP mRNA expression.Results There was a significant difference in the proliferation-inhibition rates [0.5 g/L Matrine group:(15.84 ± 2.58)％,1.0 g/L Matrine group:(23.13 ±4.19)％,1.5 g/L Matrine group:(30.32 ±3.02)％,and the control group:(8.92 ± 1.23)％];apoptotic rates [0.5 g/L Matrine group:(12.33 ± 1.15)％,1.0 g/L Matrine group:(16.67 ± 0.99)％,1.5 g/L Matrine group:(25.33 ± 1.91)％,and the control group:(9.47 ± 0.96)％];XIAP mRNA expression(0.5 g/L Matrine group:0.633 ± 0.046,1.0 g/L Matrine group:0.441 ± 0.055,1.5 g/L Matrine group:0.326 ± 0.065,control group:0.794 ±0.029)in RD cells among 0.5 g/L,1.0 g/L,1.5 g/L Matrine groups and the control group (F =14.15,83.37,50.57,all P ＜ 0.05).The proliferation-inhibition and apoptotic rates in RD cells were gradually increased with the increasing Matrine concentration.The expression of XIAP mRNA was significantly decreased in different Matrine groups compared with the control group,exhibiting a dose-dependent manner.Conclusions Matrine can inhibit the proliferation of RD cells and induce the apoptosis in a dose-dependent manner,which may be related to the down-regulated XIAP mRNA.

Objective The purpose of this paper is to study the relationship between blood concentration and brain tissue drug concentration by different dose of TMX chemotherapy acute lymphoblastic leukemia in mice. Methods 4 weeks, health Kun Ming mice 80: establishment acute lymphoblastic leukemia mice model,20 mice were randomly selected to take the femur bone marrow biopsy bone marrow OK for model verification; the remaining 60 acute lymphoblastic leukemia mice were allocated randomly 6 groups of 10 mice in each group, respectively A, B, C, D, E, F groups. And collected blood 0.5 ml and brain tissue 0.4 g individually at 0.5 hour in every group. We used supernatant of centrifugation blood and brain homogenate to detected drug concentration by fluorescence polarization immunoassay. Results The mean blood concentration of MTX of six groups A, B, C, D, E, F are (39.08±5.18) μmol/L, (15.86±1.02)μmol/L, (8.67± 5.43)μmol/L, (68.29±5.19)μmol/L, (29.55±6.22)μmol/L, (13.98±1.12)μmol/L, respectively. Compared the mean blood concentration of MTX of each group there are statistical significance (P<0.05). The mean concentration of MTX of six groups in brain tissue are followed by A group (1.05±0.26)μmol/L, B group (0.61±0.25)μmol/L, C group (0.48±0.25)μmol/L, D group (2.07±0.35)μmol/L, E group (1.27±0.21)μmol/L, F group (0.59±0.69)μmol/L. Compared the mean concentration of MTX of each group in brain tissue there are statistical significance (P<0.05). MTX concentration in blood and in brain tissue of correlation coefficient followed by 0.82, 0.75, 0.19, 0.81, 0.55, 0.43. Conclusion The chemotherapy acute lymphoblastic leukemia mice of HDMTX scheme, the peak of blood concentration and brain tissue drug concentration is come after injected MTX 0.5 hour, MTX 5 g/m~2 is better permeation blood-brain barrier and more easy make brain tissue drug concentration to reach effectively therapeutic concentration than MTX 3 g/m~2.

BACKGROUNDOncogenesis, development, invasion and metastasis of lung cancer are modulated by relative genes of lung cancer, and the expression, deletion or mutation of these genes are regulated by cell membrane signal transduction system and cell membrane ionic channels. The aim of this study is to explore the electrophysiological properties of human lung adenocarcinoma cell line A549 and cell proliferation affected by tetraethylammonium chloride (TEA), one potassium channel blocker, so as to know whether the voltage-gated potassium channels are required for the proliferation of A549 cell line.

METHODSIonic currents were recorded by whole-cell patch-clamp recording technique. The proliferation activity of A549 cells was measured by MTT assay.

RESULTSMembrane current was observed when cells were held at -70mV and test potentials ranged from -30 to +110mV. The current exhibited properties of voltage-dependent, outward rectification and no or little inactivation over the 500ms voltage pulse. Exposure of tumor cells to 10mmol/L TEA reduced the peak outward potassium current (evoked by depolarization to +110mV) from (1057.52±59.17)pA to (212.26±11.96)pA, the ratio of suppression was 79.92% (P < 0.01). Obvious inhibitive effect of TEA with different concentrations ranged from 20 to 60mmol/L on proliferation activity of the cells was found.

CONCLUSIONSThe voltage-gated potassium channels exist in human lung adenocarcinoma cell line A549 and play a great role in proliferation of A549 cells. TEA can inhibit proliferation of A549 cell through blocking the potassium channels and the inhibition is dose-dependent.

Objective:To discuss the synergistic inhibitory effect of apatinib mesylate(apatinib)combined with radiotherapy(RT)on the hepatocellular carcinoma(HepG2)cells in vitro,and to clarify its related antitumor mechanism.Methods:The HepG2 cells were cultured in vitro and treated with different concentrations of apatinib and/or varying doses of X-rays.MTT method was used to detect the survival rates of the cells in various groups;the inhibitory rates of cell proliferation and the 20％inhibitory concentration(IC20)of apatinib were calculated;the X-ray irradiation dose for subsequent experiments was detected.The HepG2 cells were divided into apatinib group,RT group,and apatinib+RT group(combined group).Flow cytometry was used to detect the apoptotic rates of the cells in various groups;wound healing assay was used to detect the migration rates of the cells in various groups;ELISA method was used to detect the levels of vascular endothelial growth factor(VEGF)in the cell culture supernatant in various groups.Results:The MTT results showed that the IC20 of apatinib was 1.32 μmol·L-1,and this concentration was used for subsequent experiments,and the X-ray irradiation dose for the follow-up experiments was 2 Gy.Compared with control group,the apoptotic rates of the cells in apatinib group and RT group had no significant differences(P＞0.05),while the apoptotic rate of the cells in combined group was increased(P＜0.05).Compared with control group,the migration rates of the cells in apatinib group,RT group,and combined group were decreased(P＜0.05);compared with apatinib group and RT group,the migration rate of the cells in combined group was decreased(P＜0.05).Compared with control group,the levels of VEGF in the cell culture supernatant in apatinib group and combined group were decreased(P＜0.05);compared with apatinib and RT group,the level of VEGF in the cell culture supernatant in combined group was decreased(P＜0.05).Conclusion:Apatinib combined with radiotherapy significantly inhibits the proliferation and migration of the HepG2 cells in vitro and induces the apoptosis;its effect may be related to the inhibition of VEGF secretion by cells.

Objective: To investigate the therapeutic effects and pharmacological mechanisms of Yishen Xingyang capsule (YXC) in oligoasthenospermia (OA) rats.Methods: Forty-eight male Sprague-Dawley rats were randomly divided into eight groups of six rats each:normal control (NC);model control (MC);three different positive drug (PD);and low-, medium-, and high-dose YXC groups. A rat model of OA was established by administering glucosides of Triptery-gium wilfordii Hook. F (GTW). After YXC administration, penile erectile function was observed. The epididymis, blood, and testes of the rats were harvested for analysis of sperm quality, sex hormone levels, mitochondrial membrane potential, and the transforming growth factor (TGF)-β1/Smad signaling pathway. Results: Compared with that in the MC group, penile erectile function in the YXC groups and three PD groups increased (all P<.01). Moreover, sperm quality in the YXC groups and three PD groups improved (all P < .001). The levels of testosterone, follicle stimulating hormone, and luteinizing hormone in the three PD and YXC groups increased (all P<.05). The mitochondrial membrane potential in the three PD and YXC groups significantly improved (all P<.001). Furthermore, the YXC and three PD groups showed decreased TGF-β1 expression (all P< .05) compared with the MC group. The high-dose YXC group and three PD groups improved Smad2 and Smad4 expression (all P<.05). Conclusion: YXC improved penile erectile function and sperm quality in OA rats, and the underlying mechanism included increase in sex hormones, inhibition of sperm apoptosis, and regulation of the TGF-β1/Smad signaling pathway. Meanwhile, this study provides a new effective drug option for the treat-ment of OA, which is beneficial to male reproductive health and social harmony.

Objective To investigate the effect of atractylone on the viability and apoptosis of hepatoma HepG2 cells and its mechanism of action. Methods Hepatoma HepG2 cells were selected and divided into low-, middle-, and high-dose atractylone groups (5, 10, and 20 μmol/L), and the cells in the control group were added with an equal volume of DMSO. MTT colorimetry was used to measure the viability of HepG2 cells after treatment with different concentrations of atractylone; flow cytometry was used to measure the apoptosis rate and mitochondrial membrane potential of HepG2 cells; the DCFH-DA fluorescent probe labeling method was used to measure the level of reactive oxygen species (ROS) in HepG2 cells; Transwell assay was used to evaluate the effect of atractylone on the migration ability of HepG2 cells; Western blot was used to measure the protein expression levels of Bcl-2, Bax, and cleaved caspase-3. A one-way analysis of variance was used for comparison of continuous data between multiple groups, and the least significant difference t -test was used for comparison between two groups. Results After 24 and 48 hours of treatment with atractylone, compared with the control group, the low-, middle-, and high-dose atractylone groups had a tendency of reduction in cell viability (all P < 0.05), with a half inhibitory concentration of 26.19 μmol/L in atractylone treatment of HepG2 cells for 72 hours. The low-, middle-, and high-dose atractylone groups had a significantly higher apoptosis rate than the control group (14.34%/29.32%/50.12% vs 0.32%, all P < 0.05). Compared with the control group, the low-, middle-, and high-dose atractylone groups had a significant increase in the fluorescence intensity of ROS in HepG2 cells (all P < 0.05). After 48 hours of treatment with atractylone, compared with the control group, the low-, middle-, and high-dose atractylone groups had a significant reduction in the number of migrated cells (132.67±18.36/57.00±9.26/31.00±2.45 vs 258.11±38.54, P < 0.05). Compared with the control group, the low-, middle-, and high-dose atractylone groups had a significant reduction in the expression of the anti-apoptotic factor Bcl-2 and significant increases in the expression of the apoptotic factors Bax and cleaved caspase-3 (all P < 0.05). Conclusion Atractylone can induce the apoptosis and inhibit the migration of HepG2 cells, which provides an experimental basis for further development and utilization of atractylone.

【Objective】 To evaluate the dietary quality with the dietary balance index (DBI_16) and the association between dietary quality and bone mass among middle-aged and elderly people in Gansu Province so as to provide evidence for improving dietary quality and bone health status of Gansu population. 【Methods】 Based on the information of the type and quantity of food intake and the bone mass of middle-aged and elderly people aged 35 years and above collected by the Gansu Project in the Regional Ethnic Cohort Study in Northwest China, DBI_16 was used to evaluate the intake level of cereals, vegetables, fruits, milk, beans, fish and shrimp, eggs and other foods, and the degree of inadequate, excessive and unbalanced dietary intake of the participants. Multiple linear regression was used to evaluate the associations of three component indexes of DBI_16, high bound score (DBI_HBS), low bound score (DBI_LBS), diet quality distance (DBI_DQD), and seven single indexes of DBI_16 with bone mass. 【Results】 Analyses of the dietary and bone mass data of 11,840 participants showed that 44.8% of participants consumed excessive amounts of cereals compared to the dietary recommendation. 96.3%, 90.6%, 90.1%, 71.9%, 95.1% and 60.3% of participants’ intake of vegetables, fruits, milk, soybeans, fish and shrimp, and eggs, respectively, were inadequate. 47.7% participants consumed less than 10 types of food. 2.3% participants’ DBI_LBS levels were appropriate. 54.7% participants’ DBI_HBS levels were appropriate. Only 1.2% participants’ DBI_DQD reached a balanced level. The bone mass level in the study population was (2.5±0.6) kg [(2.8±0.5) kg for men and (2.3±0.5) kg for women]. After adjusting for sociodemographic characteristics, lifestyle, total dietary energy intake and body mass index, DBI_LBS and DBI_DQD were negatively associated with bone mass [β and 95% CI was -0.002 01 (-0.003 62--0.000 40) and -0.001 76 (-0.003 09--0.000 43), respectively]. 【Conclusion】 Dietary intake imbalance is common among middle-aged and elderly people in Gansu Province, and the more severe the dietary intake imbalance, the lower the bone mass level.