Effects of CD73 on proliferation and function of rat cardiomyocytes in vitro / 解剖学报

Objective To investigate the effect of CD73 on the proliferation and function of rat cardiac myocytes (CMs) in vitro. Methods The primary rat CMs and H9C2 cells were cultured in vitro, and lentivirus CD73 overexpression vector and empty vector group were constructed and transfected into CMs and H9C2 respectively. Experimental groups: CD73 overexpression group (OE group) including CMs-OE group and CMs-NC group, and negative control group (NC group) including H9C2-OE grop and H9C2-NC group, 5 in each group. After transfection, the expression of CD73 gene was detected by Real-time PCR method, and the proliferation ability was detected by MTT method, and the proliferation curve, doubling time and pulsatility of CMs were detected by non-destructive cardiomyocyte function analyzer. Results After 72 hours of lentivirus transfection, CD73 mRNA level in the over expression group was significantly higher than that in the control group (P<0. 05); the proliferation ability of CMs and H9C2 at 3-4 days after transfection in OE group was larger than that in NC group (P<0. 05); the proliferation curve of CMs and H9C2 cells in OE group was higher than that in NC group, and the doubling time of CMs and H9C2 in OE group was lower than that in NC group 72 hours after CMs transfection, the beating rate of OE group was higher than that of NC group, the expression of T-box 5(TBX5) and the secretion of vasuular endothelial growth factor VEGF) in OE group were higher than those in NC group, while the secretion of hypoxia iducible factor-1α(HIF-1α) and tumor necrosis factor-α(TNF-α) was lower than that of NC group. Conclusion CD73 can promote the proliferation of CMs and H9C2, promote CMs pulsation, TBX5 expression and VEGF secretion, and inhibit the secretion of HIF-α and TNF-α.

Enhanced photodynamic therapy of TiO2/N-succinyl-chitosan composite for killing cancer cells

Abstract The aim of this study was to investigate the effect of TiO2/N-succinyl-chitosan composite (TiO2/ NSCS) photodynamic therapy (PDT), while considering the effects of various light sources on the activation of photosensitizer. The methyl thiazolyl tetrazolium assay was used to examine the cell survival rate of the cells. The results showed that glioma cell strain (U251) was the most sensitive cancer cell strain to TiO2/NSCS. When the concentration of TiO2/NSCS was between 0 and 800 μg·mL-1, there was no obvious cytotoxicity to normal liver cells (HL-7702) and U251 cells. During the PDT process, the photokilling effect of TiO2/NSCS on U251 cells under ultraviolet-A (UVA) light irradiation was stronger than that of pure TiO2, and its killing effects were positively correlated with concentration and irradiation time. In addition, both UVA and visible light could excite TiO2/ NSCS, which had significant killing effect on U251 cells. The results of acridine orange/ethidium bromide fluorescent double staining and Annexin V/propidium iodide double staining indicated that TiO2/NSCS under UVA and visible light irradiation could kill U251 cells by inducing apoptosis, and the apoptosis rate of TiO2/NSCS treatment groups was higher than that of TiO2 treatment groups. Therefore, TiO2/NSCS might be used as a potential photosensitizer in PDT.

Insights into forsythia honeysuckle (Lianhuaqingwen) capsules: A Chinese herbal medicine repurposed for COVID-19 pandemic.

Background: In December 2019, a novel coronavirus, SARS-CoV-2 caused a series of acute atypical respiratory diseases worldwide. However, there is still a lack of drugs with clear curative effects, and the clinical trial research of vaccines has not been completely finished. Purpose: LH capsules are approved TCM patent medicine that are widely used for the treatment of respiratory tract infectious diseases caused by colds and flu. On April 12, 2020, LH capsules and granules were officially repurposed by the China Food and Drug Administration (CFDA) for patients with mild COVID-19 based on their safety and efficacy demonstrated through multicentre, randomized, controlled clinical trials. We hope to conduct a comprehensive review of it through modern pharmacy methods, and try to explain its possible mechanism. Methods: Using the full names of LH capsules Lianhuaqingwen, Lianhua Qingwen andSARS-COV-2, COVID-19 as the keywords of the search terms, systemically search for existing related papers in various databases such as Web of Science and PubMed. And completed the collection of clinical data in ClinicalTrials.gov and Chinese Clinical Trial Registry. Last but not least, we have sorted out the anti-inflammatory and antiviral mechanisms of LH capsules through literature and Selleck. Results: This review systematically sorted out the active ingredients in LH capsules. Furthermore, the related pharmacological and clinical trials of LH capsule on SARS-CoV-2, IAV and IBV were discussed in detail. Moreover, the present review provides the first summary of the potential molecular mechanism of specific substances in LH capsules involved in resistance to SARS-COV-2 infection and the inhibition of cytokine storm syndrome (CSS) caused by IL-6. Conclusion: This review summarizes the available reports and evidence that support the use of LH capsules as potential drug candidates for the prevention and treatment of COVID-19. However, TCM exerts its effects through multiple targets and multiple pathways, and LH capsules are not an exception. Therefore, the relevant mechanisms need to be further improved and experimentally verified.

Preparation of Chitosan/Alginate-ellagic Acid Sustained-release Microspheres and their Inhibition of Preadipocyte Adipogenic Differentiation.

OBJECTIVE: In this study, chitosan/alginate-ellagic acid sustained-release microspheres were prepared, and the effect of sustained-release microspheres on preadipocyte adipogenic differentiation was analyzed. METHODS: Chitosan/alginate-ellagic acid microspheres were prepared and identified by scanning electron microscopy (SEM) and infrared spectroscopy (IR). The drug release rates were measured at pH 6.8, 7.0, 7.2, 7.4 to determine sustained release of ellagic acid from microspheres. The effects of 0.1, 1, 10 mg/L chitosan/alginate-ellagic acid microsphere on 3T3-F442A preadipocyte proliferation were determined by Methyl thiazolyl tetrazolium assay (MTT), and cell morphology was checked by hematoxylin/ eosin staining (HE staining). The effect of chitosan/alginate-ellagic acid microspheres on preadipocyte adipogenic differentiation was also determined by Oil red O staining, and lipogenesis was measured by isopropanol extraction. The molecular mechanism was investigated by detecting the mRNA expression of CCAAT/enhancer binding protein alpha (C/EBPα) and peroxisome proliferatorsactivated receptor gamma (PPARγ). RESULTS: Chitosan/alginate-ellagic acid sustained-release microspheres were successfully prepared, and the inhibition of proliferation and adipogenic differentiation of preadipocytes was found to be dosedependent. The mechanism of differentiation inhibition was found to be closely related to the expression of transcription factor C/EBPα and PPARγ. CONCLUSION: Chitosan/alginate can be used as a good material to prepare ellagic acid sustained-release microspheres, and these microspheres can be used for treating the obesity.

Comparative evaluation of radiopacity and cytotoxicity of platelet-rich fibrin, platelet-rich fibrin + 50wt% nano-hydroxyapatite, platelet-rich fibrin + 50wt% dentin chips: An in vitro study.

AIMS: The aim of this study is to impart radiopacity to platelet-rich fibrin (PRF) using two different bioactive agents (nano-hydroxyapatite [nHA] and dentin chips [DC]) and to assess the cell proliferation rate induced by the modified PRF. SUBJECTS AND MATERIALS: PRF was modified with 50wt% of nHA (G bone-SHAG31, Surgiwear Company) and 50wt% of DC. The five samples of each group (Group 1 - dentin disc, Group 2 - PRF, Group 3 - PRF + 50wt% nHA, and Group 4-PRF + 50wt% DC) were digitally radiographed together with 8-mm aluminum stepwedge using DIGORA software (for Windows 2.9.113.490). The aluminum equivalent of radiopacity of the samples was compared with the dentin disc (control). Further cytotoxicity (on L929 mouse fibroblast cell lines) among the groups was assessed using methyl thiazolyl tetrazolium assay. STATISTICAL ANALYSIS USED: One-way ANOVA with posthoc Tukey-honestly significant difference tests were applied to assess the significance among the various groups. RESULTS: The mean aluminum equivalent radiopacity among each group showed statistically significant results with P < 0.05. Group 3 (PRF + 50wt% nHA) can achieve an aluminum equivalent radiopacity (1.51 ± 0.089) better than Group 4 (0.97 ± 0.22 mmAl). The cell viability was >73% for all groups. CONCLUSIONS: This study found that the addition of bioactive radiopacifiers into PRF was able to impart radiopacity and these biomaterials were proved to be noncytotoxic.

Role of JAK-STAT pathway in reducing cardiomyocytes hypoxia/reoxygenation injury induced by S1P postconditioning.

This experiment was designed to explore the protection of sphingosine1-phosphate (S1P) postconditioning on rat myocardial cells injured by hypoxia/reoxygenation acting via the Janus kinase-signal transducer and activator of transcription (JAK-STAT) signal pathway. The data showed that S1P could significantly increase cell viability, lower the rate of apoptosis, decrease the content of lactate dehydrogenase (LDH) and caspase3 activity in the culture medium, increase the activity of total superoxide dismutase (T-SOD) and manganese superoxide dismutase (Mn-SOD), reduce the loss of mitochondrial membrane potential and the fluorescence intensity of intracellular calcium, as well as increase the phosphorylation of JAK2 and STAT3 in comparison with the H/R group. When the JAK inhibitor AG490 or the STAT inhibitor stattic were added, the effects of S1P were inhibited. Our date shows that S1P protects H9c2 cells from hypoxia/reoxygenation injury and that the protection by S1P was inhibited by AG490 and stattic. Therefore S1P protects H9c2 cells against hypoxia/reoxygenation injury via the JAK-STAT pathway.

Positive skeletal effect of two ingredients of Psoralea corylifolia L. on estrogen deficiency-induced osteoporosis and the possible mechanisms of action.

Estrogen replacement therapy (ERT) is utilized as a major regime for treatment of postmenopausal osteoporosis at present. However, long-term supplement of estrogen may cause uterine hyperplasia and hypertension leading to a high risk of endometrial cancer and breast cancer. Psoralea corylifolia L. has long been used as tonic and food additives in many countries. Previous studies had found two ingredients in P. corylifolia L.: bavachin and bakuchiol exhibited osteoblastic activity. The present study was designed to investigate the protective effect of bakuchiol and bavachin on ovariectomy-induced bone loss and explore the possible mechanism. In vivo, bakuchiol and bavachin could prevented estrogen deficiency-induced bone loss in ovariectomized rats without uterotrophic activity. In vitro studies suggested that bakuchiol and bavachin induced primary human osteoblast differentiation by up-regulating the Wnt signalling pathway. This study suggests that such a bone-protective role makes them a promising and safe estrogen supplement for the ERT.

Preparation of intravenous injection nanoformulation of VESylated gemcitabine by co-assembly with TPGS and its anti-tumor activity in pancreatic tumor-bearing mice.

Our recent publication showed that VES-dFdC nanocapsules in pure water could be obtained via the self-assembling of VES-dFdC prodrug synthesized by coupling gemcitabine (dFdC) with vitamin E succinate (VES). To prepare the intravenous injection nanoformulation, we present here a novel strategy to improve the stability and drug concentration of VES-dFdC nanoformulation in PBS or isotonic solution. Particularly, D-α-tocopheryl polyethylene glycol succinate (TPGS), usually used as drug solubilizer and coincidentally contains the same VES moiety as VES-dFdC prodrug and PEG chain, is selected to co-assemble with VES-dFdC prodrug. The zeta potentials of all the TPGS/VES-dFdC co-assemblies were close to 0 mV, and their particle size measured by dynamic light scattering (DLS) decreased from 113 to 36 nm with increasing TPGS/VES-dFdC molar ratios from 0.15 to 1.5. Stable colloidal suspensions were obtained without aggregates in PBS at 4 °C in one month or isotonic solution at 37 °C in one week, and the weight concentration of VES-dFdC prodrug increased from 7 to 17 mg/mL when the molar ratios of TPGS/VES-dFdC ranged from 0.5/1 to 1.5/1. The concentration of VES-dFdC prodrug was high enough to be used as intravenous injection nanoformulation in nude mice. Interestingly, along with the increase of TPGS/VES-dFdC molar ratios from 0.3/1 to 1.5/1, the morphology of TPGS/VES-dFdC co-assemblies changed from loose nanocapsule to compact micelle revealed by transmission electron microscope (TEM). Finally, the co-assembly of TPGS/VES-dFdC (TPGS/VES-dFdC: 1/1) was selected as intravenous injection nanoformulation to evaluate the antitumor activity. Compared with native dFdC, TPGS/VES-dFdC nanoformulation with 0.2mmol/kg of dosage showed similar low toxicity in vivo, but 4.7 times high of tumor inhibition rate in nude mice with pre-established BxPC-3 tumors.

Study on the anaphylactoid of three phenolic acids in Honeysuckle.

ETHNOPHARMACOLOGICAL RELEVANCE: Chlorogenic acid (CA) is one of the major ingredients in Honeysuckle which exhibits anticancer, antibacterial, antiviral, hypoglycemic and anti-HIV activities. However, with the frequent emergence of anaphylactoid reactions of traditional Chinese medicine (TCM) injections which contains Honeysuckle in recent years, many researchers found that CA exhibited allergenicity. AIM OF THE STUDY: To explore the changes of content of CA, neochlorogenic acid (NCA) and cryptochlorogenic acid (CCA) during the preparation of Honeysuckle and evaluate the anaphylactoid of these three ingredients. MATERIALS AND METHODS: Two extracts of Honeysuckle were prepared by different methods and the content of CA, NCA and CCA were determined by high performance liquid chromatography (HPLC) analysis. Guinea pig and mast cells RBL-2H3 were utilized as the animal and cell model to investigate the anaphylactoid of these three ingredients. RESULTS: The content of CA decreased while CCA and NCA increased during the preparation of Honeysuckle. In vitro and in vivo studies showed that CA and CCA could significantly increase the plasma serotonin and ß-hexosaminidase levels in guinea pigs and induce the degranulation of RBL-2H3 cell, while NCA did not show such properties. In addition, CAA had a stronger effect than CA. CONCLUSION: We believed that both CA and CCA could cause anaphylactoid reaction while NCA could not. Moreover, the anaphylactoid of CCA is higher than CA. Our result demonstrated that CA is not the only cause of anaphylactoid reactions of TCM injections which contains Honeysuckle.

Musashi2 modulates K562 leukemic cell proliferation and apoptosis involving the MAPK pathway.

The RNA-binding protein Musashi2 (Msi2) has been identified as a master regulator within a variety of stem cell populations via the regulation of translational gene expression. A recent study has suggested that Msi2 is strongly expressed in leukemic cells of acute myeloid leukemia patients, and elevated Msi2 is associated with poor prognosis. However, the potential role of Msi2 in leukemogenesis is still not well understood. Here, we investigated the effect of Msi2 knockdown on the biological properties of leukemic cells. High expression of Msi2 was found in K562 and KG-1a leukemic cell lines, and low expression was observed in the U937 cell line. We transduced K562 cells with two independent adenoviral shRNA vectors targeting Msi2 and confirmed knockdown of Msi2 at the mRNA and protein levels. Msi2 silencing inhibited cell growth and caused cell cycle arrest by increasing the expression of p21 and decreasing the expression of cyclin D1 and cdk2. In addition, knockdown of Msi2 promoted cellular apoptosis via the upregulation of Bax and downregulation of Bcl-2 expression. Furthermore, Msi2 knockdown resulted in the inactivation of the ERK/MAPK and p38/MAPK pathways, but no remarkable change in p-AKT was observed. These data provide evidence that Msi2 plays an important role in leukemogenesis involving the MAPK signaling pathway, which indicates that Msi2 may be a novel target for leukemia treatment.

Cardioprotection of 17ß-estradiol against hypoxia/reoxygenation in cardiomyocytes is partly through up-regulation of CRH receptor type 2.

Estrogens have been suggested to exert cardioprotection through maintaining endogenous cardioprotective mechanisms. In the present study, we investigated whether estrogens protect cardiomyocytes against hypoxia/reoxygenation (H/R) via modulating urocortins (UCNs) and their receptor corticotrophin-releasing hormone receptor type 2 (CRHR2). We found that 17ß-estradiol (E2) enhanced UCN cardioprotection against H/R and increased CRHR2 expression in neonatal rat cardiomyocytes. E2 protected cardiomyocytes against H/R, which was impaired by CRHR2 antagonist or knockdown of CRHR2. Estrogen receptor α (ERα) antagonist treatment or ERα knockdown could abolish E2-induced CRHR2 up-regulation. Moreover, knockdown of Sp1 also attenuated E2-induced CRHR2 up-regulation. Ovariectomy resulted in down-regulation of CRHR2 and Sp-1 in myocardium of mice, which was restored by E2 or ERα agonist treatment. These results suggest that estrogens act on ERα to up-regulate CRHR2 expression in cardiomyocytes, thereby enhancing cardioprotection of UCNs against H/R.

Enhancement of expression of survivin promoter-driven CD/TK double suicide genes by the nuclear matrix attachment region in transgenic gastric cancer cells.

This work aimed to study a novel transgenic expression system of the CD/TK double suicide genes enhanced by the nuclear matrix attachment region (MAR) for gene therapy. The recombinant vector pMS-CD/TK containing the MAR-survivin promoter-CD/TK cassette was developed and transfected into human gastric cancer SGC-7901 cells. Expression of the CD/TK genes was detected by quantitative real-time PCR (qPCR) and Western blot. Cell viability and apoptosis were measured using the methyl thiazolyl tetrazolium (MTT) assay and flow cytometry. When the MAR fragment was inserted into the upstream of the survivin promoter, the qPCR result showed that the expression of the CD/TK genes significantly increased 7.7-fold in the transgenic SGC-7901 cells with plasmid pMS-CD/TK compared with that without MAR. MTT and flow cytometry analyses indicated that treatment with the prodrugs (5-FC+GCV) significantly decreased the cellular survival rate and enhanced the cellular apoptosis in the SGC-7901 cells. The expression of the CD/TK double suicide genes driven by the survivin promoter can be enhanced by the MAR fragment in human gastric cancer cells.

Brucine, an alkaloid from seeds of Strychnos nux-vomica Linn., represses hepatocellular carcinoma cell migration and metastasis: the role of hypoxia inducible factor 1 pathway.

Brucine is an alkaloid derived from the seeds of Strychnos nux-vomica Linn. which have long been used as a traditional medicine for the treatment of hepatocellular carcinoma (HCC) in China. HCC prognosis can be greatly influenced by metastasis. There has thus far been little research into brucine as a source of anti-metastasis activity against HCC. In this study, we revealed that brucine dramatically repressed HepG2 and SMMC-7721 HCC cell migration with few cytotoxic effects. Hypoxia inducible factor 1 (HIF-1) is a key transcription factor mediating cell migration and invasion. Brucine suppressed HIF-1-dependent luciferase activity in HepG2 cells. The transcriptions of four known HIF-1 target genes involved in HCC metastasis, i.e., fibronectin, matrix metallopeptidase 2, lysyl oxidase, and cathepsin D, were also attenuated after brucine treatment. Experiments in vivo showed that an intraperitoneal injection of 5 and 15 mg/kg of brucine resulted in dose-dependent decreases in the lung metastasis of H22 ascitic hepatoma cells. Moreover, a dosage of brucine at 15 mg/kg exhibited very low toxic effects to tumor-bearing mice. Consistently, brucine downregulated expression levels of HIF-1 responsive genes in vivo. Our current study demonstrated the capacity of brucine in suppressing HCC cell migration in vitro and lung metastasis in vivo. The inhibition of the HIF-1 pathway is implicated in the anti-metastasis activity of brucine.

Design, synthesis and biological evaluation of novel mansonone E derivatives prepared via CuAAC click chemistry as topoisomerase II inhibitors.

Two series of novel C-9 chloro- and bromo-substituted mansonone E derivatives with triazole moieties at the C-3 position were prepared by using copper-catalysed azide-alkyne cycloaddition click chemistry. These compounds were found as potent inhibitors of topoisomerase II (Topo II) and topoisomerase I (Topo I). The Topo II-mediated pBR322 DNA relaxation and cleavage assay showed that the derivatives might act as catalytic inhibitors. Their cytotoxic activities against A549, HL-60, K562 and HeLa cells were evaluated, indicating that these compounds were potent antitumour agents. Their structure activity relationships and molecular docking study revealed that the substituents of the triazole were particularly important for cytotoxicity.

Role of glycosylation in the anticancer activity of antibacterial peptides against breast cancer cells.

Antibacterial peptides (ABPs) with cancer-selective toxicity have received much more attention as alternative chemotherapeutic agents in recent years. However, the basis of their anticancer activity remains unclear. The modification of cell surface glycosylation is a characteristic of cancer cells. The present study investigated the effect of glycosylation, in particular sialic acid, on the anticancer activity of ABPs. We showed that aurein 1.2, buforin IIb and BMAP-28m exhibited selective cytotoxicity toward MX-1 and MCF-7 breast cancer cells. The binding activity, cytotoxicity and apoptotic activity of ABPs were enhanced by the presence of O-, N-glycoproteins, gangliosides and sialic acid on the surface of breast cancer cells. Among N-, O-glycoproteins and ganglioside, O-glycoproteins almost had the strongest effect on the binding and cytotoxicity of the three peptides. Further, up-regulation of hST6Gal1 in CHO-K1 cells enhanced the susceptibility of cells to these peptides. Finally, the growth of MX-1 xenograft tumors in mice was significantly suppressed by buforin IIb treatment, which was associated with induction of apoptosis and inhibition of vascularization. These data demonstrate that the three peptides bind to breast cancer cells via an interaction with surface O-, N-glycoproteins and gangliosides. Sialic acids act as key glycan binding sites for cationic ABP binding to glycoproteins and gangliosides. Therefore, glycosylation in breast cancer cells plays an important role in the anticancer activity of ABPs, which may partly explain their cancer-selective toxicity. Anticancer ABPs with cancer-selective cytotoxicity will be promising candidates for anticancer therapy in the future.

Syntheses and evaluation of novel isoliquiritigenin derivatives as potential dual inhibitors for amyloid-beta aggregation and 5-lipoxygenase.

A series of new isoliquiritigenin (ISL) derivatives were synthesized and evaluated as dual inhibitors for amyloid-beta (Aß) aggregation and 5-lipoxygenase (5-LO). It was found that all these synthetic compounds inhibited Aß (1-42) aggregation effectively with their IC50 values ranged from 2.2 ± 1.5 µM to 23.8 ± 2.0 µM. These derivatives also showed inhibitory activity to 5-LO with their IC50 values ranged from 6.1 ± 0.1 µM to 35.9 ± 0.3 µM. Their structure-activity relationships (SAR) and mechanisms of inhibitions were studied. This study provided potentially important information for further development of ISL derivatives as multifunctional agents for Alzheimer's disease (AD) treatment.

Effect of dietary nucleotides on immune function in Balb/C mice.

This study investigated the effect of dietary nucleotides (NTs) on immune function in female Balb/C mice, which randomly distributed into six groups: one control group, one NF-free (NF) control group and four NT groups. NTs ranged from 0.0025% to 0.64%. Compared with the control group, the NF could significantly weaken the activity of T lymphocytes and macrophages, as well as decreased the activity of B lymphocytes and NK cell. NF significantly decreased the ratio of CD4(+)/CD8(+), whereas, it increased Tr percentage. In comparison with the NF group, the concentration of serum IL-2 and IL-4 showed an increase trend. Meanwhile, the granular cell macrophages colony stimulating factor (GM-CSF) increased significantly in the 0.04% NT group. The ratio of Th1/Th2 also showed an increasing trend after the supplements of NTs. There were no significant differences between the control and 0.04% NT group. Nevertheless, no significant differences in weight gain and lymphoid organ indices were observed in our study. These results indicate that NT supplements can prevent hypoimmunity which result from NF diet. 0.04% NTs is the healthy optimal supply proportion in mice diet.

Comparison of adherence ability of bacteria on the surface of rigid gas-permeable contact lens made with different materials / 中华实验眼科杂志

Background Wearing contaclenincreasethe risk of infection of the cornea.Some studieshowed the gas-permeability of materialused foconstructing corneal contaclenione of the contributing factorrelated to corneal health.Objective Thistudy wato observe the in vitro adherence ability of differenbacterito rigid gas-permeable contaclense(RGP-CL) made with varioumaterials.MethodContaclensemade with hexafocon,enflufocon opolymethyl methacrylate (PMMA) were placed into Staphylococcuaureus,Staphylococcuepidermidis,oPseudomonaaeruginosbacterial suspension(0.5 MCF) fo24 hours.The strength of bacterial adherence watested and studied by the methyl thiazolyl tetrazolium (MTT) colorimetrimethod based on absorbance (value),and the vortex method waused to calculate the colony forming units.The bactericlump formation waexamined with scanning electron microscope (SEM).ResultMTcolorimetrimethod showed thathe adherence ability of Staphylococcuaureuto hexafocon (value) wasignificantly lowethan thato enflufocon and PMMA,respectively (q=7.379,8.207,P＜0.01),buno significandifference wafound in the adherence ability of Staphylococcuaureubetween enflufocon and PMM(q =0.828,P＞0.05).The adherence ability of Staphylococcuepidermidito XO and enflufocon walowethan thato PMM(q =14.000,12.800,P＜0.01),buno significandifference wafound between the adherence of Staphylococcuepidermidito hexafocon and enflufocon material (q =1.200,P＞0.05).There wano significandifference in the adherence ability of Pseudomonaaeruginosto all three material(F=2.155,P=0.138).The vortex method presented the colony forming unitof Staphylococcuaureuto hexafocon,enflufocon and PMMwith (37.9± 1.5)×106,(49.9±2.2)×106 and (67.4± 1.6)×106,respectively,with significandifference among them (F =206.240,P＜0.01),showing the lowesvalue in hexafocon,the highesvalue in PMMand middle value in enflufocon (q=11.650,28.640,16.990,P＜0.01),Moreover,colony forming uniof Staphylococcuepidermidito hexafocon,enflufocon and PMMwa(7.9 ± 1.3) × 106,(10.5 ± 1.5) × 106,(11.2 ±1.2) × 106,respectively.And thaof hexafocon walowethan one of the PMMmaterial (q =5.060,P＜0.05).No significandifference wafound between hexafocon and enflufocon nobetween hexafocon and PMM(q =3.290,1.770,P＞0.05).In addition,the resultthacorresponded to the vortex method were seen in the MTcolorimetriassay (F =0.232,P =0.799).SEM examination showed dispersed population of Staphylococcuaureuand Staphylococcuepidermidion the surfaceof hexafocon and enflufocon;while much more Staphylococcuaureuand Staphylococcuepidermidiadhered on the surface of PMMA,forming net-like appearance.Conversely,high numbeof Pseudomonaaeruginoswaseen on the surface of all three materials,withounoticeable differencein the bacterial shape and quantity on each of the material.ConclusionThe adherence ability of bacterito PMMistrongethan thaof hexafocon and enflufocon,and gas-permeable material of RGP-CL doenoimpacthe adherence ability of bacteria.

Effects of Aurora A silence by RNA interference on the apoptosis and proliferation of glioma cells / 解剖学报

Objective To investigate the inhibitory effect of RNA interference on the expression of Aurora A in U251 cells, and the influence on proliferation and apoptosis of U251 cells. Methods The siRNA specific for Aurora A was synthesized and transfected into U251 cells in vitro. Aurora A mRNA expression and protein content were detected by RT-PCR and Western blotting respectively. The cell proliferation and apoptosis were observed by methyl thiazolyl tetrazolium(MTT) and flow cytometry(FCM). Transmission electron microscope was used to observe the ultrastructural changes of U251 cells. Results After transfection, the expression level of Aurora A mRNA was significantly decreased(P<0.01), and the protein content of Aurora A was also obviously reduced. The inhibitory rate of cell proliferation reached up to 67.57% 72 hours after transfection, which was significantly higer than that of normal control group(P<0.01). The apoptosis rate of U251 cells was significantly increased from (3.69±0.87)% to (15.34±2.16)% (P<0.01). Under the transmission electron microscope, it was observed that the U251 cells showed typical morphologic changes of apoptosis after transfection, such as karyopyknosis, chromatin condensation and margination, intracytoplasmic vacuoles formed, and apoptotic bodies formed. Conclusion The expression of Aurora A gene can be inhibited by siRNA successfully, and it results in the suppression of cell growth and induce apoptosis of human glioma cells in vitro. Aurora A may become a new target for the gene therapy of gliomas.

HSP40 and HSP70 inhibit the accumulation and toxicity of mutant huntingtin in N2a cells / 解剖学报

Objective To study the effect of heat shock protein 40(HSP40) and heat shock protein 70(HSP70) on the aggregate formation of mutant huntingtin (htt) and its toxicity in N2a cells. Methods The effect of the over-expression of HSP40 and HSP70 alone or co-over-expression of them on aggregate formation of transfected mutant htt (containing 150 glutamine repeats, 150Q) in N2a cells was detected by fluorescence microcopy and Western blotting. Cell viabilities were detected by MTT assay. Reactive oxygen species (ROS) production was detected by colorimetric method. Results The over-expression of HSP40 and HSP70 alone, especially the co-over-expression of them, dramatically decreases the formation of 150Q htt aggregates in N2a cells. The numbers of aggregates in each group are as follows ( n = 1 000): about 50% in the group of 150Q htt-expressing alone, about 12% in the group of over-expression of HSP40, about 15% in the group of over-expression of HSP70, about 5% in the group of co-over-expression of HSP40 and HSP70. The result of MTT assay shows that the over-expression of HSP40 and HSP70 alone, especially the co-over-expression of them, dramatically increases cell viabilities ( P<0.01, n =3) and reduces the production of ROS ( P<0.01, n =3). Conclusion HSP40 and HSP70 can increase cell viabilities of 150Q N2a cells via inhibiting the aggregates formation of mutant huntingtin and reducing oxidative stress.