目的 探讨制痂酊对大鼠深Ⅱ度烧伤创面修复及可能作用机制.方法 SD 100只大鼠,制备深Ⅱ度烧伤大鼠模型,随机分为5组:正常组、模型组、阳性对照组(湿润烧伤膏)、制痂酊高剂量组、制痂酊低剂量组.给药组每日换药,连续给药14 d.烧伤后3、7、14 d检测创面愈合率和创面含水量;生化分析法检测创面组织超氧化物歧化酶(SOD)、过氧化氢酶(CAT)活性及丙二醛(MDA)含量;测定大鼠血清肿瘤坏死因子-α(TNF-α)、白介素-1β(IL-1 B)、白介素-6(IL-6)的水平.结果 制痂酊可不同程度地提高大鼠烧伤创面愈合率,降低烫伤创面组织含水量(P<0.05);明显升高烧伤大鼠SOD的活性,下调MDA的含量(P<0.05),降低烧伤大鼠血清TNF-α、IL-1β、IL-6含量(P<0.05,P<0.01).结论 制痂酊能够提高创面愈合率和SOD活性,减少创面含水量及MDA含量,降低血清炎症介质TNF-α、IL-1β、IL-6的水平,对深II度烧伤大鼠的创伤有一定的修复作用.

<span id="ChDivSummary" name="ChDivSummary" class="abstract-text">目的探讨凋亡诱导因子p53抑制HT22细胞（小鼠海马神经元细胞）谷氨酸毒性的相关机制。方法 CCK-8法和PI/Hoechst荧光双染法检测HT22细胞存活率;Western blot检测p53以及胱氨酸/谷氨酸反向转运系统xCT（cystine/glutamate antiporter或system Xc^-, xCT或SLC7A11）蛋白表达;DHE荧光探针检测HT22细胞内活性氧簇（reactive oxygen species, ROS）;BODIPY 581/591 C11脂质氧化探针和4-HNE免疫荧光染色,检测细胞内脂质氧化;FeRhoNox^TM-1荧光探针检测细胞内铁离子。结果 1μmol·L^-1 Tenovin-1作用6 h后,p53蛋白的表达水平相较于对照组明显上升。在p53高表达后,经谷氨酸盐（glutamate, Glu）和铁死亡诱导剂Erastin处理8 h后,相较于仅Glu和Erastin处理组细胞死亡率明显下降,xCT蛋白表达水平明显上升。p53高表达后的Glu处理组细胞内ROS、脂质氧化以及Fe²⁺的水平明显低于单纯Glu处理组。结论 p53可能通过调节xCT表达来抑制铁死亡,进而保护神经不受谷氨酸的损伤。</span>

[目的]调查古井园国家级自然保护区药用维管植物的资源种类与分布特点.[方法]通过实地调查、标本采集与鉴定,整理与分析调查结果.[结果]古井园国家级自然保护区分布的药用维管植物278种.[结论]古井园国家级自然保护区药用维管植物资源丰富,应可持续利用野生中药资源和合理发展中药种植,调整农业产业结构,促进中药产业的发展.

目的:探讨RAB32对慢性粒细胞白血病(CML)K562细胞增殖和迁移作用的影响.方法:使用Western blot和qRT-PCR分别检测RAB32在CML患者、健康对照以及CML K562细胞株中的表达情况.进一步转染RAB32-shRNA到K562细胞株中敲除RAB32基因后,流式细胞仪分析K562细胞周期变化,Western blot和qRT-PCR分别检测K562细胞中细胞增殖相关蛋白C-myc、Cyclin D1和细胞迁移相关蛋白MMP-3、MMP-9的蛋白和mRNA表达水平变化.结果:Western blot和qRT-PCR检测结果显示,RAB32在CML患者和K562细胞株中高表达(P<0.01).在K562细胞株中转染RAB32-shRNA抑制RAB32表达后,细胞周期分析结果显示,较NC-shRNA组相比,RAB32-shRNA组K562细胞的S期和G2/M期细胞比例均有不同程度降低,且S期细胞比例降低更为显著(P<0.01).Western blot检测结果显示,与NC-shRNA组相比,RAB32-shRNA组K562细胞的细胞增殖相关蛋白C-myc、Cyclin D1和细胞迁移相关蛋白MMP-3、MMP-9蛋白表达水平明显降低(P<0.01),qRT-PCR检测结果显示,这四种因子的mRNA水平也相应降低(P<0.01).结论:抑制RAB32的表达能够抑制CML K562细胞的增殖和迁移能力.

目的:分析貌似前庭周围性病变的小脑梗死的临床和影像学特点及临床定位错误的原因.方法:连续3年观察发病1周内收治的小脑梗死患者107例,所有患者均由发病1周内的MR1检查确诊并按TOAST分型.所有患者均行MRA或CTA评估颅内血管情况,行颈部血管彩色超声评估颈部血管情况.其中有11例临床表现貌似前庭周围性病变.比较貌似前庭周围疾病的小脑梗死患者(观察组)和一般小脑梗死患者(对照组1)及同期收治的27例前庭周围性眩晕患者(对照组2)的一般资料和危险因素.结果:11例观察组的病灶均位于小脑半球,在小脑后下动脉及小脑前下动脉的供血分布区,病灶直径均＜2 cm,病灶成点状、条状或片状.观察组与对照组1比较,除脑梗死病史外,其他的一般资料和危险因素差异无统计学意义(P＞0.05);观察组与对照组2比较,二者之间除房颤外,性别、年龄、高血压史、糖尿病史、高脂血症病史、高同型半胱氨酸血症病史、冠心病痛史、吸烟史及既往卒中史的差异均有统计学意义(P＜0.05).行logistic回归分析显示:年龄、高血压、糖尿病、卒中史为貌似前庭周围疾病的小脑梗死的独立危险因素.结论:早期行头MR检查有助于明确诊断貌似前庭周围疾病的小脑梗死,对于高龄、既往有脑梗死、糖尿病、高血压的患者出现发作性头晕、耳鸣等临床表现,其病灶可能在小脑.

Objective To investigate the influence of bcl-xL overexpression in donor endothelia on recipient immunoreaction after rat aorta transplantation.Methods Enhanced green fluorescent protein (EGFP)-marked lentivirus express vector with Tie2 promoter of anti-apoptotic gene bcl-xL was con structed and transfected into donor SD rat by tail vein injection.The isogeneic and allogeneic aorta transplantation was performed between SD and SD rats as well as SD and Wistar rats.Mter one week of trans plantation,the overexpression of bcl-xL gene in transplant aorta was detected by fluorescence microscope and real-time quantitative polymerase chain reaction (Real-time PCR).The serum from recipients was collected and transplant aortic endothelial cells,spleen cells as well as cytotoxic T lymphocytes (CTL) from donor and recipient were separated and cultured.The mixed lymphocyte reaction,CTL activity and complement-dependent antibody-mediated cytotoxicity were then performed to assess the influence of bcl-xL overexpression in donor endothelia on recipient immunoreaction after rat aorta transplantation.Results Bcl-xL gene was significantly overexpressed (8.5 ± 1.1 vs.1.0 ± 0.2,t =17.050,P =0.002) in transplant aorta after transfection of lentivirus express vector with Tie2 promoter.The prolifera tion rate of recipient spleen cells stimulated by allogeneic donor spleen cells was significantly increased in contrast to stimulated by isogeneic donor spleen cells (14.2 ± 1.8 vs.1.0 ±0.1,t =23.599,P =0.001;14.7 ±2.2 vs.0.8 ±0.1,t =25.037,P =0.001).The survival rate of transplant aortic endothelial cells incubated by allogeneic recipient CTL and serum was significantly reduced in contrast to incubated by isogeneic recipient CTL and serum (14.5％ vs.92.7％,/x2 =244.558,P =0.000;8.9％ vs.95.8％,x2 =290.494,P=0.000;15.1％ vs.96.7％,x2=269.251,P=0.000;10.1％ vs.94.7％,x2=286.190,P =0.000).However,there were no significant differences of the proliferation rate (1.0 ±0.1 vs.0.8±0.1,t=2.108,P=0.068;14.2±1.8vs.14.7±2.2,t=-1.012,P=0.341) and survival rate (92.7％ vs.96.7％,x2 =3.078,P=0.079;95.8％ vs.94.7％,x2 =0.497,P=0.481;14.5％vs.8.9％,x2=2.917,P=0.088;15.1％ vs.10.1％,x2=2.286,P=0.131;and 96.6％ vs.97.3％,x2 =0.344,P=0.558;91.5％ vs.92.1％,x2 =0.033,P=0.856;19.8％ vs.15.4％,x2 =0.387,P =0.239;10.2％ vs.13.8％,x2 =1.515,P =0.218) between transfected groups and non transfected groups in both isogeneic and allogeneic transplantation.Conclusion No significant effect of bel-xL overexpression in donor endothelia on recipient immunoreaction after rat aorta transplantation.

Objective To study the anti-hepatofibrotic effects of Zhizi Bopi Decoction and its compatibility on liver fibrosis. Methods The model of mouse liver fibrosis which was induced by carbon tetraehlofide ( CCl4 ) was used to investigate the effects of Zhizi Bopi Decoction and its compatibility by gavage. Indicators of hepatic fibrosis and inflammation were detected with automated biochemistry analyzer and liver fibrosis of the mice was assessed by Mas-son staining and HE staining. Meanwhile, the protein levels of collagen typeⅠ and α-smooth muscle actin (α-SMA) were evaluated by Western blot analysis. Results Zhizi Bopi Decoction and each different treatment group could evidently inhibit the increased serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), hydroxyproline ( Hyp) and hyaluronic acid ( HA) activities, and improve the pathological changes of mouse livers. Furthermore, the therapeutic effects on liver fibrosis induced by CCl4 in mice were primarily ascribed to the syner-gic effect of Gardenia jasminoides Ellis, Cortex phellodendri and Glycyrrhizae Radix et Rhizoma ( P<0. 01 ) . And they could obviously reduce the protein expression levels of collagen typeⅠandα-SMA in the liver ( P<0. 01 ) . In addition, Zhizi Bopi Decoction could also obviously reduce the contents of transforming growth factor-beta ( TGF-β) . Conclusion Zhizi Bopi Decoction and its compatibility all perform significant anti-fibrotic effects on the liver in different degree, but the compatibility of Gardenia jasminoides Ellis, Cortex phellodendri and Glycyrrhizae Radix et Rhizoma exert the most distinctly anti-fibrotic effects among them, possibly by reducing the contents of TGF-β.

目的 研究环耙明在巨噬细胞极化过程中的作用.方法 用100 ng/ml脂多糖(LPS)和20 ng/ml干扰素γ(IFN-γ)处理RAW264.7 24 h刺激成M1型巨噬细胞,用20ng/ml白介素-4(IL-4)处理RAW264.7 24 h刺激成M2型巨噬细胞,用荧光定量PCR(QPCR)法检测各分型中一氧化氮合成酶(iNOS)、CD86、精氨酸酶-1(Arg-1)、CD206、GLi1、ptch1 mRNA水平的表达;用QPCR法、Western blot法、免疫荧光法检测加入环耙明后对M1型巨噬细胞分泌iNOS的影响.结果 M1型巨噬细胞高分泌iNOS、CD86、M2型巨噬细胞高分泌Arg-1、CD206(P＜0.01);40 nmol/L环耙明刺激后,M1型巨噬细胞中ptch1 mRNA水平的表达明显增强且在100 nmol/L时达到最大值(P＜0.01);经环耙明1 000nmol/L刺激后有效降低了iNOS的mRNA和蛋白水平,可能提示环耙明会促进巨噬细胞向M2型巨噬细胞分化.结论 环耙明能够明显地降低M1型巨噬细胞中iNOS的分泌.

Objective To explore effect and molecular mechanism of improved insulin resistance (IR) in HepG2 cells by tanshinone Ⅰ.Methods IR-HepG2 was induced by high concentrations of insulin .The appropriate con-centrations of tanshinone Ⅰ were determined by MTT assay.The glucose consumption was detected by glucose as -say kit.The expressions of protein tyrosine phosphatase 1B(PTP1B), P-AKT, AKT were determined by Western blot.Results The IR-HepG2 cells were established successfully by incubating with 10-7 mol /L insulin for 24 hours.The ultimate concentrations of tanshinone Ⅰ were determined as 78, 156, 312 ng /ml and tanshinone Ⅰconcentration-dependently increased glucose consumption of IR -HepG2 cells.Western blot showed increased ex-pression of PTP1B and decreased expression of P -AKT in model group compared with the normal group .Moreover, tanshinone Ⅰ concentration-dependently decreased expression of PTP1B and increased expression of P-AKT com-pared with the model group.Conclusion Tanshinone Ⅰ improved IR in HepG2 cells.The putative mechanism is that tanshinone Ⅰ decrease expression of PTP1B and promotes activation of P -AKT/AKT signaling pathway.

Objective To investigate the expression profile of voltage-gated potassium channel Kv1. 3 in lipopo-lysaccharide(LPS)-induced acute liver injury. Methods C57BL/6 mice were injected with LPS intraperitoneally for 24 h,to establish the model of acute liver injury. The morphological changes of liver tissues were observed by HE staining. Alanine aminotransferase ( ALT) and aspartate aminotransferase ( AST) levels were measured using test kits. The Kv1. 3 expression was measured by immunohistochemistry, Florescent real-time quantitative RT-PCR ( qRT-PCR) and Western blot. Kupffer cells ( KCs) were isolated by in situ perfusion, and then applied to detect Kv1. 3, tumor necrosis factor alpha(TNF-α) expression by qRT-PCR. The expression of Kv1. 3 was also detected in LPS-stimulated RAW264. 7 cells. Margatoxin( MgTx) was pretreated to block Kv1. 3 in LPS-stimulated RAW264. 7 cells, then detected the expression of cytokines by qRT-PCR and Western blot. Results HE staining showed that liver tissueswere damaged by LPS,ALT and AST levels increased significantly after LPS treatment(P<0. 01). The Kv1. 3 expression decreased significantly in model group compared to normal group(P<0. 01). Kv1. 3 was low ex-pressed in KCs in LPS-inducedacute liver injury model group ( P <0. 01 ) . LPS-stimulated RAW264. 7 cells ex-pressedlow level of Kv1. 3(P<0. 05), and MgTx, the Kv1. 3 specific blocker, decreased LPS-induced cytokynesis secretion. Conclusion The results indicate that Kv1. 3 is low expressed in LPS-induced acute liver injury and bloc-king of the Kv1. 3 channel decreases LPS-induced cytokynesis secretion in macrophage. Kv1. 3 may play an impor-tant role in the treatment of liver diseases.

Objective: To study changes in expression of claudin-11 and proteins related to mitogen-activated protein kinase (MAPK) signaling pathways, as well as the ultrastructure of the blood testis barrier (BTB), in male ICR mice exposed to decabromodiphenyl ether (BDE-209). Methods: Fifty-two mice, 4 weeks of age, weighing 15-21 g, were provided with adaptive feeding for 1 week. Mice were randomly divided into 4 groups, named control, low-dose, medium-dose and high-dose groups. The treated groups received BDE-209, by intragastric gavage, at doses, respectively, of 100, 300 and 500 mg/kg. Mice were sacrificed after 6 weeks and organs harvested on ice, weighed and stored at -80 °C. The ultrastructure of testicular tissues was examined by electron microscopy. Western blotting was used to detect proteins related to the MAPK pathway, including p38 mitogen activated protein kinase (p38), phosphorylated p38 (p-p38), extracellular regulated protein kinase 1/2 (ERK1/2) , phosphorylated ERK1/2 (p-ERK1/2) , c-jun N-terminal kinase (JNK), phosphorylated JNK (p-JNK) and the BTB tight junction protein claudin-11. Analyze the difference between each groups. Results: At sacrifice, the body weights in each treated group were compared with those in the control group weighing (41.14 ± 0.60) g. Compared with controls, body weights were significantly different (P<0.05) in the middle dose, at (39.97 ± 0.66) g and high dose, at (39.98± 0.55) g in control group. The coefficients of the testis were significantly lower (P<0.05) in each treated group than in controls, with values of (0.37±0.0)%, (0.31±0.05)% and (0.31±0.04)% for low-dose, medium-dose and high-dose groups, respectively. The epidymus coefficient values were also significantly lower than controls (P<0.05), with values of (0.16±0.06)%, (0.11±0.05)% and (0.07±0.03)%, respectively in the same three dose groups. Electron microscopy ultrastructure showed that, compared with the control group, the testes in the middle and high dose groups had closely connected fractures, cell edema and more vacuoles. Compared with in the control group, levels of p-p38 and p-JNK in testicular tissue were significantly increased (P<0.05). In the control group and in low-, medium- and high-dose groups, the p-p38/p38 ratios were 1.35±0.13, 3.46±0.10, 5.71±0.26 and 4.79±0.21, respectively. The corresponding p-JNK/JNK ratios were 2.07±0.0, 4.77±0.18, 3.63±0.06 and 4.85±0.15. Claudin-11 levels were significantly lower (P<0.05) than control values in each dosed group. The corresponding values in control, low-dose, medium-dose and high-dose groups were 8.33±0.36, 2.06±0.27, 3.37±0.27 and 1.55±0.19, respectively. Conclusion: BDE-209 increased expression of proteins in the MAPK pathway and decreased expression of the BTB tight junction protein claudin-11 in testicular tissue. It also caused ultrastructural damage to the Sertoli cell BTB tight junctions. This suggested that BDE-209 might damage Sertoli cells BTB through effects on the MAPK pathway.

Objective To screen out the derivative with better anti-inflammatory activity of Hesperidin. And its an-ti-inflammatory activity was also evaluated then. Methods We stimulated RAW264. 7 cell strains with lipopolysac-charide ( LPS) and then inflammation cytokines tumor necrosis factor alpha ( TNF-α) and interleukin 6 ( IL-6 ) were measured by ELISA. And the anti-inflammatory activity was evaluated on three inflammation models( mice ear swelling induced by xylol, mice foot swelling induced by carrageen glue and mice adjuvant-induced arthritis. Re-sults Derivatives of hesperidin 12 ( HY-12 ) had better anti-inflammatory activity; HY-12 ( 100 , 200 , 400 mg/kg) on acute inflammation induced by xylene in mice auricle had different degree of inhibition, HY-12 (200, 400 mg/kg) dose group could significantly inhibit the ear swelling in mice;HY-12 (100 mg/kg) in 3 h, 5 h had obvi-ously inhibitory effect on mice foot swelling, HY-12(200,400 mg/kg) in inflammatory started producing curative effect after 1 h, at all time points it was significantly inhibited. HY-12 also had certain inhibition to inflammation of rheumatoid arthritis mouse model. HY-12 400 mg/kg could significantly inhibit the secondary side paw swelling, serum inflammatory factors, local inflammatory infiltration and the expression of TNF-αand IL-6 in synovial tissues of adjuvant-induced arthritis mice. And nuclear factor kappaB p65 played gets a major role in regulation of inflam-mation response. Conclusion The above results show that HY-12 has strong anti-inflammatory activity through regulating the activation of NF-κB.

Objective To investigate the expression changes of histone deacetylases ( HDACs) levels in alcohol-in-duced liver cell injury in mice. Methods Mice liver cell line (AML12) was used as the research object in vitro, to determine alcohol inhibitory effect on AML12 cells at different concentrations and time points by using MTT method;flow cytometry anlaysis was used to determine alcohol induced apoptosis effect on AML12 cells. Real-time quantitative PCR analysis was used to detect mRNA expression levels of HDACs in alcohol-induced liver cell injury model. Results Upon the stimulation of alcohol concentration at 100 mmol/L for 24 h, the survival rate of AML12 cell was 84%,indicating its significant role in inhibiting the proliferation of liver cell line(AML12) (P<0. 05);furthermore, the apoptosis rate of AML12 cells was 16%,showing it could increase the AML12 cell apoptosis( P<0. 05 ) . Binge alcohol exposure induced liver cell injury and eventually resulted in deregulation of hepatic HDACs mRNA expressions. It could be observed visually that HDAC1,2,3,4,5,6,7,8 mRNA expressions were signifi-cantly down-regulated and HDAC9 mRNA expression was up-regulated(P<0. 05). However, there was no signifi-cant difference in HDAC10 mRNA expression than control. Conclusion In alcohol-induced liver cell injury model, alcohol consumption affects HDACs mRNA levels and therefore, we speculate that histone deacetylases-mediated epigenetic modifications may play an important role in the pathogenesis of alcohol-induced hepatic injury.

Objective To study the protective effects of the inhibiting of HY-12 to CYP2E1 on CCl4 induced acute liver injury in mice. Methods All 70 kunming mice were randomly divided into control group, model group, HY-12 high, medium and low dose group and positive controldrug( glycyrrhizic acid group and hesperidin group) . Mice were given 0. 5% sodium carboxymethycellulose and the corresponding drug for 7 days. At the end of the seventh&nbsp;day, the mice were injected with 0. 2% CCl4 (0. 1 ml/10 g) to induce mouse model of acute liver injury except the-normal group. To observe the level of liver homogenate about serum alanine aminotransferase (ALT), aspartate aminotransferase ( AST) , and the influence of the malondialdehyde ( MDA) , superoxide dismutase ( SOD) , gluta-thione peroxidase (GSH-px) level;liver tissue has been stained by HE. The expression of CYP2E1 in liver tissue was measured by immunohistochemistry and Western blot. The expression level of CYP2E1 mRNA in the mice liver was detected by qRT-PCR. Results HY-12 could decrease ALT,AST activity in mice serum of the acute liver in-jury,reducethe level of MDA in liver homogenate, increase SOD and GSH-px levels, inhibit the CYP2E1 level in liver cells, and significantly reduce the degree of liver tissue lesions in mice. Conclusion HY-12 has certain pro-tective effect on acute liver injury in mice and its mechanism may be related to the body lipid peroxidated and in-hibited CYP2E1 expression.

Objective To investigate the effect of geniposide on regulating the proliferation and activation of PDGF-induced HSC-T6 cells. Methods HSC-T6 cells were cultivated by geniposide with different concentrations (0, 20, 50, 100, 200, 400 μg/ml), and or after the cells were stimulated with PDGF, respectively. Cell activity, mRNA and total protein expressions were assayed by MTT, qRT-PCR and Western blot. Results The geniposide pretreatment effectively inhibited PDGF-mediated proliferation and the expressions of the activation markers (α-SMA) in HSC-T6, and also inhibited the phosphorylation levels of Akt,mTOR and p70S6K. But the geniposide couldn′t affect the activation levels of ERK and p38. Furthermore,the protein of the MAPK pathway ( P-ERK and P-p38) and Akt/mTOR/p70S6K signaling pathway were detected by Western blot. Conclusion The proliferation and activation of HSC-T6 induced by PDGF are inhibited by geniposide treatment, and then it maybe provide new i-deas and targets for the prevention of liver fibrosis, which is probably through modulating the Akt/mTOR/p70S6K signaling pathway.

Aim To investigate the effects of cell pro-liferation and activation in HSC-T6 cells by inhibiting the expression of EZH2 , and its partial relevant mech-anism. Methods By introducing the inhibitor DZNep in activated HSC-T6 cells stimulated by TGF-β1 , the protein expression levels of EZH2, p-ERK, p-AKT andα-SMA were detected by Western blot. The siRNA targeting EZH2 was designed and synthesized according to its nucleotide sequence, and their corresponding ex-pression vectors were constructed and transfected into HSC-T6 cells with LipofectamineTM 2000. The prolifer-ation of HSC-T6 cells was determined by MTT. And the protein expression levels of EZH2, p-ERK, p-AKT and α-SMA were measured by Western blot. Results&nbsp;By introducing the inhibitor DZNep in activated HSC-T6 cells stimulated by TGF-β1 , it effectively de-creased the protein levels of EZH2 and also the protein levels of p-ERK, p-AKT and α-SMA. By introducing EZH2-siRNA in activated HSC-T6 cells, it effectively inhibited the cell proliferation, and also the protein levels of EZH2, p-ERK, p-AKT andα-SMA. Conclu-sion Silencing EZH2 expression inhibits HSC-T6 cell proliferation and activation, and EZH2 may be a poten-tial therapeutic target gene for hepatic fibrosis.

The fibrosis can occur in many kinds of organs,and its sustained progress may lead to organ structural damage and functional decline,and even the organ failure,which threatens the human health and the life seriously.Adenosine is an endogenous purine nucleoside that can be generated in various tissues of the body and regulate a multitude of body functions via the combina-tion with four different kinds of G protein-coupled receptors.Re-cent studies have found that adenosine receptors play an impor-tant role in regeneration tissue and fibrosis process.To under-stand the processes may be helpful to the treatment of fibrosis diseases.This review makes a summary on latest research pro-gress of adenosine receptors in fibrosis diseases.

Aim To observe the effect of melittin on human hepatocelluar carcinoma HepG2 cell prolifera-tion in vitro and its further mechanisms.Methods The capacity of cellular proliferation and apoptosis was measured with the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay,Hoechst 33258 assay and Annexin V-FITC /PI assay.The mR-NA expression of Shh, PTCH1, SMO, GLi1 and HDAC2 was performed by qRT-PCR.And the protein expression of Shh,PTCH1,SMO,GLi1 and HDAC2 was assessed by western blotting.Results Our study found that melittin effectively inhibited cell prolifera-tion and promoted cell apoptosis in vitro using MTT method and Flow cytometry.The mRNA and protein expression of Shh,PTCH1,SMO,GLi1 and HDAC2 were obviously decreased after treated with various con-centrations of melittin for 48h in HepG2 cells.Conclu-sions Taken together,our data suggest that melittin could inhibit cell proliferation and promote cell apopto-sis,reduce the level of HDAC2 and down-regulate the Hedgehog signaling pathway in this process simultane-ously.

Objective To investigate the inhibitory effect of novel compound containing dihydropyrazole moiety on the proliferation of SMMC-7721 by targeting hTERT melittin. Methods The inhibition rate of proliferation of cells treated with novel compounds was measured by MTT assay. And the novel compounds’ effect on cell cycle was test-ed by flow cytometry cell cycle experiment. Telomerase activity was determined through modified Telomeric Repeat Amplification Protocol (TRAP) assays. The protein expression level of hTERT was observed by Western blot. Re-sults Compared with the heterocyclic compounds, novel compound containing dihydropyrazole moiety had obvious inhibitory effect on the proliferation of cell lines, especially on the human hepatoma cell line SMMC-7721. The re-sult of flow cytometric cell cycle analysis showed that the number of cells in S phase was markedly increased. After the treatment with novel compound containing dihydropyrazole moiety, the results of modified TRAP assays predic-ted that the telomerase activity was inhibited. And the results of western blot showed that the protein expression level of hTERT decreased obviously. Conclusion Novel compound containing dihydropyrazole moiety inhibits the prolif-eration of SMMC-7721 and makes them arrested at S phase. This initial discovery is mainly achieved through the inhibition of telomerase activity by targeting hTERT.