Pien Tze Huang (片仔癀)-induced apoptosis in human colon cancer HT-29 cells is associated with regulation of the Bcl-2 family and activation of caspase 3

Objective  

To investigate the cellular effects of Pien Tze Huang (片仔癀, PZH) in the HT-29 human colon carcinoma cell line.     

Xihuang Pill (西黄丸) Induces Mesenchymal-Epithelial Transition and Inhibits Loss of Apical-Basal Polarity in Colorectal Cancer Cell through Regulating ZEB1-SCRIB Loop

Objective：To investigate the antiproliferative and anti-metastasis effect of Xihuang Pill（西黄丸,XP） on human colorectal cancer cell and to explore the molecular mechanism by which it produces the effects.Methods：Highly metastatic human colorectal cancer cell line LoVo was treated with low-,medium-,and highdose XP-containing serum（XP-L,XP-M,XP-H） groups for 48 h,cells intervened with no drug rat serum and PD98059[extracellular signal-regulated kinase（ERK） inhibitor]as negative and positive controls（NC and PC）groups.Cell proliferation assay was made using cell counting kit-8（CCK8）.The 8 μm pore-size transwell chamber and 4＇,6-diamidino-2-phenylindole（DAPI） staining were applied to examine the ability of invasion and migration of the cells.The protein expression of ERK1/2,zinc finger E-box-binding homeobox 1（ZEB1）,Scrib and lethal giant larvae homolog 2（Lgl2） was detected by Western blotting while the relative mRNA quantity of E-cadherin,N-cadherin,Occludin and junctional adhesion molecule-1（JAM1） was measured by realtime fluorescent quantitative polymerase chain reaction（RT-qPCR）.Results：XP induced a dose-dependent suppression on the proliferation of LoVo cells（P〈0.05 or P〈0.01）,with the inhibition rates varied from 27.30%to31.08%.Transwell assay showed that when preprocessed with PD98059 and XP-containing serum,the number of cells that passed the filter decreased significantly compared with that of NC group（P〈0.05 or P〈0.01）.Moreover,XP inhibited the protein expression of ERK1/2 and ZEB1（P〈0.05）;and up-regulated the protein expression of Scrib and Lgl2（P〈0.05）.The mRNA levels of E-cadherin,Occludin and JAM1 of the XP intervened groups and PC group markedly ascended（P〈0.05） while that of N-cadherin showed a descending tendency（P〉0.05）.Conclusion：XP intervention suppressed the ability of proliferation,invasion and migration of the LoVo cells.Regulating ZEB1-SCRIB Loop so as to recover epithelial phenotype and apical juncti     

Qushi Huayu Decoction (祛湿化瘀方) Inhibits Protein and Gene Expression of Cathepsin B in HepG2 Cells Induced by Free Fatty Acids

Objective: To study the experimental efficacy of Qushi Huayu Decoction (祛湿化瘀方,QHD) on protein and gene expression of cathepsin B (ctsb) in HepG2 cells induced by free fatty acids (FFAs).Methods: The model of HepG2 steatosis and tumor necrosis factor-α (TNF-α) secretion was induced by long-chain FFAs.HepG2 cells were divided into 4 groups: control group (group C),model group (group M),low-dose QHD group (group L) and high-dose QHD group (group H ).Long-chain FFAs were added to groups M,L and H.The 10% blank-control serum was added to group C and M,while 5% and 10% QHD-containing sera were added to group L and H,respectively.The levels of serum TNF-α and cellular triglyceride (TG) were detected.Cellular p-IκB and ctsb expression were detected using Western blot and PCR.The expression and distribution of ctsb were observed by immunofluorescence.Results: After incubating with FFA for 24 h,TG deposition in HepG2,TNF-α content in cell supernatant,the protein expression of cellular ctsb and P-IκB,as well as mRNA expression of ctsb increased markedly in group M compared with group C (P0.05,P0.01).Compared with group M,TG deposition,the expression of cellular ctsb,P-IκB and ctsb mRNA in groups L and H,as well as TNF-α content in group H,decreased significantly (P0.05).Cell immunochemical fluorescence studies showed that ctsb was released from lysosomes and distributed in the cytoplasm extensively and diffusedly after being stimulated with FFA.In this study,these above-mentioned changes were inhibited markedly in groups L and H.Conclusion: QHD might have a direct inhibitory effect on the ctsb target in the FFA-ctsb-TNFα pathway of hepatic lipotoxicity.     

Effects of Zao Huang Mixture (藻黄合剂) on the Expressions of TGF-β1 and Col IV in Human Glomerular Mesangial Cells Cultured in High Glucose Environment

Objective: To investigate the effect of Zao Huang Mixture (藻黄合剂ZHM) on expressions of growth factor-β1 (TGF-β1) and collagen IV (Col IV) in human glomerular mesangial cells (GMC) cultured in high-glucose environment. Methods: After primary culture of GMC, in vitro culture was carried out in normal group, high glucose group and high glucose medium with ZHM of different concentrations, and the expressions of TGF-β1 and Col IV in the GMC group and in ZHM group were detected at 24 and 48 h respectively. Results...     

钙调素拮抗剂O-(4-乙氧基)-丁基-小檗胺对乳腺癌多药耐药细胞系MCF-7/ADR的耐药逆转作用

目的研究钙调素拮抗剂O-4-乙氧基-丁基-小檗胺(EBB)对乳腺癌多药耐药细胞系MCF-7/ADR的耐药逆转作用。方法采用四唑盐(MTT)比色法测定药物敏感性,分析联合应用低剂量EBB(≤IC20)时阿霉素(ADR)对MCF-7/ADR细胞的半数抑制浓度(IC50)值及其增敏倍数;应用流式细胞术测定EBB对细胞周期的影响,同时观察细胞内药物浓度的积累;采用逆转录—多聚酶链反应检测EBB对多药耐药基因-1(mdr1)及拓扑异构酶Ⅱb(topⅡb)mRNA表达水平的影响。结果EBB对MCF-7/ADR的多药耐药具有明显逆转作用,3、7·5μmol/L浓度的EBB可以明显提高MCF-7/ADR对ADR的敏感性,平均增敏倍数分别为50·40、89·80倍,逆转强度明显高于阳性逆转剂维拉帕米(VPL)的14·88倍(P=0·0097)。6μmol/L的EBB处理2h后,细胞内积累的P糖蛋白底物罗丹明123(Rh123)荧光强度峰值发生明显右移;同时EBB可明显增强ADR对MCF-7/ADR细胞在G2/M期的阻滞作用。6和12μmol/L的EBB处理MCF-7/ADR细胞48h后,mdr1的mRNA表达水平有一定的降低趋势,但无统计学差异;topⅡb基因表达差异亦无显著性。结论EBB对MCF-7/ADR细胞具有较强的逆转多药耐药作用,其可能的逆转机制在于增强ADR对耐药细胞在G2/M期的阻滞以及抑制P糖蛋白外排泵作用。     

Clinical Study on Zhenjian Granule (针箭颗粒) in Improving Essential Hypertension and Insulin Resistance

Clinical Study on Zhenjian Granule (针箭颗粒) in Improving Essential Hypertension and Insulin Resistance@唐蜀华$Cardiovascular Department, Affiliated Hospital of Nanjing University of TCM!Nanjing (210029) @蒋卫民$Cardiovascular Department, Affiliated Hospital of Nanjing University of TCM!Nanjing (210029) @陈晓虎$Cardiovascular Department, Affiliated Hospital of Nanjing University of TCM!Nanjing (210029)…     

Effects and potential mechanisms of Danzhi Xiaoyao Pill (丹栀逍遥丸) on proliferation of MCF-7 human breast cancer cells in vitro

Objective： To investigate the effects of 50% ethyl alcohol （EtOH） extracts from Danzhi Xiaoyao Pill （丹栀逍遥丸, DXP） on the proliferation of MCF-7 human breast cancer cells and potential mechanisms. Methods： ATP-Lite assay was performed to test the proliferation of the MCF-7 breast cancer cell line; and antioxidant activity was measured by the oxygen radical absorbance capacity （ORAC）. The effects of DXP on nitric oxide （NO） production were tested by lipopolysaccharide （LPS）- stimulated RAW 264.7 murine macrophages using the Griess reaction. Results： The 50% EtOH DXP extracts displayed a cytotoxic response on MCF-7 cells at 0.10, 0.25 and 0.50 mg/mL dosedependently with the proliferation inhibited by more than 85%. The ORAC value of the DXP was 820μ moL Trolox equivalent/g, about 40% of the vitamin C value. DXP extracts had significant inhibitory effect on NO production at the concentration from 0.0625 mg/mL to 0.5 mg/mL （P〈0.05, P〈0.01）. Conclusion： The extracts of DXP could significantly inhibit the proliferation of MCF-7 cells, with the effect possibly related to its antioxidant activity and the inhibition of NO production.     

Modified Xiaochaihu Decoction (加味小柴胡汤) Promotes Collagen Degradation and Inhibits Pancreatic Fibrosis in Chronic Pancreatitis Rats

Objective: To investigate the effect of Modified Xiaochaihu Decoction(MXD, 加味小柴胡汤) on collagen degradation in rats with chronic pancreatitis(CP). Methods: Rats were injected dibutyltin dichloride(DBTC, 7 mg/kg of body weight) into the right caudal vein to induce CP model. Thirty heallhy male Wistar rats were randomly divided into three groups by a random number table: the control, the model and the treatment groups. Rats of treatment group were administered MXD(10 g/kg of body weight) orally once daily starting from the day post-model establishment. Pancreatic tissues were harvested after 28-day feeding and fibrosis was evaluated by picro-sirius red staining. The contents of collagen type Ⅰ and Ⅲ were detected using enzymelinked immunosorbent assay(ELISA), the expression of matrix metalloproteinase 13(MMP13) and tissue inhibitor of metalloproteinase 1(TIMP1) was analyzed by Western blot and real-time polymerase chain reaction(PCR). Results: The fibrosis scoring of pancreatic tissues, the concentrations of collagen type Ⅰ and Ⅲ, the expression levels of MMP13 and TIMP1 proteins and mRNA in the model group were all increased compared with the control group(P0.05). After treatment with MXD, the fibrosis scoring of pancreatic tissues, the concentrations of collagen type Ⅰ and Ⅲ, the expression levels of MMP13 proteins and m RNA in the teatment group were all decreased compared with the model group(P0.05), but there were no significant differences in the expression levels of TIMP1 proteins and m RNA(P0.05). Conclusion: MXD could promote collagen degradation and reverse pancreatic fibrosis in CP rats via a mechanism involve up-regulation of MMP13 expression.     

A Comparative Study of ~(99m)Tc-YIGSR and 99mTc-MIBI Uptake in Tumor Cells

99mTc YIGSR,anewtumorimagingagentwhichcanspecificallybindtothereceptorsonthesurfaceoftumorcell,wasusedtostudythecelluptakedynamicsinEhrlichascitestumorcellsandcomparedwiththatof99mTc MIBI.1MATERIALSANDMETHODS1.1Materials1.1.1ReagentsLaminin YIGSR(SigmaCorp.,USA);99mTcO-4(ChinaScientificResearchInstitu teofAtomicEnergy);SephadexG10(PharmaciaCorp.,USA);S Acetly NH3MAG3(donatedbyProf.Hnatowich,USA);Sep pakC18reverse phasecolumn(WatersMilfortM.A.USA);Sncl2(AldrichCh…     

Effects of Compound Zhebei Granule (复方浙贝颗粒) Combined with Doxorubicin on Expression of Specific Surface Antigens in Mice with Transplanted KG-1a Cells

Objective:To investigate the effects of Compound Zhebei Granule(复方浙贝颗粒,CZG)combined with doxorubicin hydrochloride(adriamycin,ADM)on specific surface antigens in mice with KG-1a transplanted cells.Methods:A subcutaneous tumor xenograft model was established by injection of the acute myeloid leukemia cell line KG-1a into the axillary flanks of BALB/c-nude mice.Twenty-four tumor bearing mice were divided into 4groups according to a random number table,including normal saline control group,ADM group,high-dose CZG group,and mid-dose CZG group,with 6 mice in each group.Drug administration occurred on the 14th day after cell inoculation,and normal saline control group mice were gavaged with normal saline at 0.2 m L/10 g every other day.ADM group mice were intraperitoneally injected with ADM 1 mg/kg[conversion of adults,40 mg/(m~2·d)]every other day.High-and mid-dose CZG groups mice were gavaged with CZG at the dose of 8 and 4 g/kg respectively every other day and intraperitoneally injected with ADM(1 mg/kg)every other day.The administration period lasted for10 days.The tumor xenografts were made into mononuclear cell suspensions after dissection,and the expressions of specific surface antigens,including CD34~+CD38~-,CD34~+CD38~-CD123~+,CD34~+CD38~-CD96~+and CD34~+CD38~-CD33~+,in KG-1a cell line tumor xenografts were detected by flow cytometry.Results:Compared with the control and ADM groups,expression of CD34~+CD38~-in the two CZG groups was significantly lower(P0.05).Compared with the control group,expression of CD34~+CD38~-CD123~+in the two CZG groups decreased(P0.05).The high-dose CZG group showed more obvious outcomes compared with the ADM group(P0.05).Compared with the control and ADM groups,the expression of CD34~+CD38~-CD96~+and CD34~+CD38~-CD33~+in the two CZG groups decreased(P0.05).Conclusions:CZG combined with doxorubicin could reduce the expression of CD34~+CD38~-,CD34~+CD38~-CD123~+,CD34~+CD38~-CD96~+and CD34~+CD38~-CD33~+in KG-1a cell line tumor xenografts,which shows that CZG could target leukemia stem cells and play a role in chemosensitization.     

Effects of Danhong Injection(丹红注射液) and Its Main Components on Anticoagulation and Fibrinolysis in Cultured Vein Endothelial Cells

Objective: To observe the effects of Danhong Injection(丹红注射液) and its main components, including daiclzein and hydroxysafflor yellow A(HSYA), on the anticoagulation, fibrinolysis, anti-apoptosis in hypoxia model of vein endothelial cells(VECs). Methods: VECs were prepared and were put in a hypoxia environment, which consisted of mixed gas of 95% N_2 and 5% CO_2 mixed gas, when reached confluent culture. Five groups used different treatments, including normal control group, hypoxia group, daiclzein group, HSYA group and Danhong Injection group. The VECs were identified by fluorescence double labeling methods. The morphology was observed by a phase contrast microscopy. The effects of Danhong Injection, daiclzein and HSYA on 6 keto prostaglandin F1α(6-keto-PGF1α) level was measured by the method of radioimmunoassay(RIA). Superoxide dismutase(SOD) activity was tested by water soluble tetrazolium salt. The content of malondialdehyde(MDA) was measured by thiobarbituric acid. The activities of tissue-type plasminogen activator(t-PA) and plasminogen activator inhibitor(PAI) were measured by the method of chromogenic substrate. The contents of endothelin(ET) and nitric oxide(NO) were detected by non-equilibrium RIA and enzymelinked immunosorbent assay. Cells apoptosis rate was determined by flow cytometry. Results: Compared with the normal control group, the floating cells number, PAI activity, ET and MDA contents, and cells apoptosis rate in the culture solution of hypoxia group were all significantly increased, whereas the 6-keto-PGF1α and NO contents, and t-PA and SOD activities were decreased significantly(P0.01). Compared with the hypoxia group, Danhong Injection markedly increased the 6-keto-PGF1α content and SOD activity, regulated PAI and t-PA activities, ET and NO contents, and decreased MDA content and cells apoptosis rate(P0.05 or P0.01). Conclusions: Danhong Injection and its main components played an important role in protecting primary VECs from hypoxic damage by regulating the secretion and vasomotor function of VECs. The function of Danhong Injection was most remarkable.     

Guizhi Decoction (桂枝汤) Inhibits Cholinergic Transdifferentiation by Regulating Imbalance of NGF and LIF in Salt-Sensitive Hypertensive Heart Failure Rats

Objective: To observe the imbalance of anatomical and functional innervation factors of sympathetic nerves, nerve growth factor(NGF) and leukemia inhibitory factor(LIF), in salt-sensitive hypertensive heart failure rats and to explore the effects of treatment with Guizhi Decoction(桂枝汤) on sympathetic remodeling by inhibiting cholinergic transdifferentiation. Methods: SS-13 BN and Dahl salt-sensitive(DS) rats were divided into 3 groups: SS-13 BN group(control group, n=9), DS group(model group, n=9) and GS group(Guizhi Decoction, n=9). After 10 weeks of a high-salt diet, the GS group rats were given Guizhi Decoction and other two groups were given saline at an equal volume as a vehicle. After 4 weeks' intragastric administration, rats were executed to detect the relevant indicators. Echocardiography and plasma n-terminal pro-B type natriuretic peptide(NT-proBNP) levels were used to assess cardiac function. Noradrenaline(NA) levels in the plasma and myocardium were detected to evaluate the sympathetic function. NGF and LIF expression were detected in the myocardium by Western blot or quantitative real-time PCR. Double immunofluorescence or Western blot was used to detect tyrosine hydroxylase(TH), choline acetyltransferase(CHAT) and growth associated protein 43(GAP43) in order to reflect anatomical and functional changes of sympathetic nerves. Results: DS group had anatomical and functional deterioration of sympathetic nerves in the decompensation period of heart failure compared with SS-13 BN group. Compared with the DS group, Guizhi Decoction significantly decreased the expression of LIF mRNA/protein(P0.01), increased the expression of NGF(P0.05 or P0.01), enhanced the levels of TH~+/GAP43~+ and TH~+/CHAT~+ positive nerve fibers(P0.01), and improved the protein expression of TH and GAP43 in left ventricle, but had no effect on CHAT(P0.05). Guizhi Decoction inhibited inflammatory infiltration and collagen deposition of myocardial injury, increased the content of myocardial NA(P0.05), reduced the plasma NA level(P0.01), improved cardiac function(P0.01), and improved weight and blood pressure to some extent(P0.05), compared with DS group. Conclusions: Guizhi Decoction could inhibit cholinergic transdifferentiation of sympathetic nerves, improve the anatomical and functional denervation of sympathetic nerves, and delay the progression of decompensated heart failure. The mechanism may be associated with the correction of the imbalance of NGF and LIF.     

Morphological Changes in Network of Enteric Nerve-Interstitial Cells of Cajal-Smooth Muscle Cells in Rats with Multiple Organ Dysfunction Syndrome and Therapeutic Effects of Dachengqi Decoction (大承气汤)

Objective: To observe the effects of Dachengqi Decoction (大承气汤, DCQD) on morphological changes in the network of enteric nerve-interstitial cells of Cajal (ICCs)-smooth muscle cells (SMC) of enteric deep muscular plexuses (DMP) in the rats with multiple organ dysfunction syndrome (MODS). Methods: One hundred Wistar rats of both sexes weighing 200 to 250 g were randomly divided into the control group, MODS group, and DCQD group. The morphologic changes of enteric nerve-ICC-SMC network in the DMP of intestine was observed using c-Kit and vesicular acetylcholine transporter/neuronal nitric oxide synthase immunohistochemical double-staining with whole-mount preparation technique, confocal laser scanning microscopy, and electron microscopy. Results: Compared with the control group, the distribution and densities of cholinergic/nitrergic nerves and ICC in the DMP (ICC-DMP) of intestine in the MODS group were significantly decreased (P0.01), and the network of cholinergic nerve-ICC-SMC was disrupted; and the ultrastructural features of ICC-DMP, enteric nerve, and SMC were severely damaged. After treatment with DCQD, the damage in the network of enteric nerve-ICC-SMC was significantly recovered. Compared with the MODS group, the distribution and densities of cholinergic/nitrergic nerves and ICC-DMP in the DCQD group were significantly increased (P0.01); and the ultrastructural features of ICC-DMP, enteric nerve, smooth muscle cells were significantly recovered. Conclusions: DCQD can improve the gastrointestinal motility in MODS. The mechanism may be related to the effect of repairing the damages in the network of enteric nerve-ICC-SMC.     

Bioequiwalence of clavulanate Potassium and Amoxicillin(1:7) dispersible tablets in healthy volunteers

Clavulanate Potassium( CL S) and Amoxicillin( AMC) ( 1 :7) dispersible tablets,twice daily,isindicated for short term treatment of bacterial in-fections at several sites when amoxicillin resistantβ— lactamase producing strainsare suspected as thecause.Resistance to many antibiotics is caused bybacterial enzymes which destroy the antibiotic be-fore it can act on the pathogen.The ClavulanatePotassium in the compound can circumventthis de-fense mechanism by blocking theβ- lactamase en-zyme…     

The State of Insulin Resistance in Patients with Nonalcoholic Fatty Liver and the Intervention with Gankangyin(肝康饮)

Insulinsensitivityofthepatientswithnonalcoholicfattyliver(NAFL)arelow,andhyperinsulinemiaandinsulinresistance(IR)arepresentinthem(1,2).Hyperinsu linemiaandIRarethepathophysiologicba sisofmetabolicsyndromeandtheinitiatingfactorofNAFL(2).Inrecentyears,somescholarshavedonehelpfulexplorationcon cerningIRandtraditionalChinesemedicine(TCM).Theyfoundthatinpatientswithsimplehypertriglyceridemiaorhyperten sion,theconditionofhyperinsulinemiawasmoresevereinthosewithStheniasyndromethaninthosewithA…     

Phenotypic Modulation of Mesangial Cells in Diabetic Rats and Effect of Tujian Mixture (菟箭合剂) on It

Phenotypic Modulation of Mesangial Cells in Diabetic Rats and Effect of Tujian Mixture (菟箭合剂) on It@尹德海$Peking Union Medical College Hospital!Beijing (100730) @梁晓春$Peking Union Medical College Hospital!Beijing (100730) @郑法雷$Peking Union Medical College Hospital!Beijing (100730) @朴元林$Peking Union Medical College Hospital!Beijing (100730)…     

Guidelines for diagnosis and treatment of HIV/AIDS in China (2005)

Acquired immunodeficiency syndrome （AIDS）, ,caused by the human immunodeficiency virus （HIV）, has become a major public health issue in China. It not only posesses formidable challenges for the health of the Chinese people, but has also influenced China＇s economic development and social stability.     

Follicle-stimulating Hormone(FSH) Induced Internalization of Porcine FSH Receptor in Cultured Porcine Granulosa Cells and Chinese Hamster Ovary Cells Transfected with Recombinant Porcine FSH Receptor cDNA

It is now well known thatthe binding of severalprotein ligands to specific cell- surface receptors is fol-lowed by the internalization and( or) degradation ofthe bound protein.The process of internalizationcould be observed by using the methods of fluores-cent,ferritin- labeled,gold- labeled,and　 12 5I- labeledligands[1] .The results from such experiments havedelineated a pathway( called receptor- mediated endo-cytosis) thatinvolves the internalization of the boundligand via coated pits and v…