OBJECTIVETo investigate the effect of valproate acid sodium (VPA) on gap junction intercellular communication in breast cancer Hs578T cells and explore the mechanism.

METHODSMTT assay was used to detect the cytotoxicity of VPA on Hs578T cells, and parachute assay was used to detect the effect of VPA on dye spread of the cells. Western blotting was employed to detect the expression changes of Cx43 total protein in VPA-treated Hs578T cells. The effect of VPA on the expression of Cx43 on the surface of Hs578T cells was examined with immunofluorescence assay.

RESULTSMTT assay showed no obvious cytotoxicity of VPA on Hs578T cells at the concentrations below 10 mmol/L. VPA below 5 mmol/L obviously increased the gap junction function in Hs578T cells (P<0.01), and significantly enhanced the expression of Cx43 total protein (P<0.01) and Cx43 expression on the surface of Hs578T cells (P<0.01).

CONCLUSIONVPA can obviously increase the gap junction function in Hs578T cells possibly by enhancing Cx43 total protein expression and Cx43 protein expression on the surface of Hs578T cells.

Breast Neoplasms ; metabolism ; Cell Communication ; drug effects ; Cell Line, Tumor ; Connexin 43 ; metabolism ; Female ; Gap Junctions ; drug effects ; Humans ; Valproic Acid ; pharmacology

Objective To observe the analgesia efficacy of continuous fascia iliaca compartment block (FICB) and adductor canal block (ACB) on elderly patients undergoing total knee arthroplasty (TKA) with multimodal analgesia and their early rehabilitation. Methods Sixty TKA patients (26 female and 34 male, 60-75 years old) were randomly divided into two groups and received ultrasound-guided either continuous FICB (n=30) or continuous ACB (n=30) using 0.25% ropivacaine for controlled analgesia. All the patients had no previous experiences of knee surgery. Quality of analgesia was assessed by dynamic and static visual analogue scale (VAS) based on pain intensity. In addition, rehabilitation training compliance, range of motion (ROM) and knee function were assessed at different times after rehabilitation training. Results No significant differences were found in VAS scores during resting and passive functional exercise at 6, 12, 24, 48 h after TKA surgery (P>0.05); ACB group had significant differences in rehabilitation training compliance, knee ROM and HSS scores in comparison with FICB group (P<0.05). Conclusions Ultrasound-guided continuous ACB with multimodal analgesia could promote early rehabilitation after TKA．

OBJECTIVETo investigate the effect of sodium valproate, a histone deacetylase inhibitor, on the cytotoxicity of doxorubicin in breast cancer cells.

METHODSWestern blotting was used to assess Cx43 protein expression in breast cancer Hs578T cells exposed to doxorubicin and sodium valproate. MTT assay was used to determine the cytotoxicity of doxorubicin; annexin V/PI double staining and Hochest 33258 fluorescence staining were employed to detect doxorubicin-induced early and late apoptosis, respectively.

RESULTSWestern blotting showed that sodium valproate significantly increased Cx43 protein expression in Hs578T cells (P/0.01). The cells exposed to both sodium valproate and doxorubicin showed significantly lowered cell viability compared with the cells exposed to doxorubicin alone (P/0.01). Exposure to both sodium valproate and doxorubicin resulted in significantly increased early and late cell apoptosis rate compared with doxorubicin treatment alone (P/0.01).

CONCLUSIONsodium valproate can significantly enhance the cytotoxicity of doxorubicin and increase doxorubicin-induced apoptosis in breast cancer cells in vitro possibly by enhancing the gap junction function.

Apoptosis ; drug effects ; Breast Neoplasms ; pathology ; Cell Line, Tumor ; drug effects ; Cell Survival ; drug effects ; Connexin 43 ; metabolism ; Doxorubicin ; pharmacology ; Drug Synergism ; Gap Junctions ; Histone Deacetylase Inhibitors ; pharmacology ; Humans ; Valproic Acid ; pharmacology

OBJECTIVETo investigate the effect of Src kinase inhibitor PP2 on intercellular communication of gap junction in breast cancer cells.

METHODSCultured breast cancer Hs578T cells were treated with various concentrations of pp2 (0,1,2,4,8,16,32 μmol/L) for 24h. Cell growth was determined by MTT assay; dye spread in Hs578T cells was measured by Parachute assay; and the expression of Src kinase in Hs578T cells was detected by Western blot.

RESULTSMTT assay showed that the survive rate of Hs578T cells treated with PP2 (1 ≊ 8 μmol/L) was 98% ± 3% ≊ 94 % ± 4%. Parachute assay showed that compared to control group the standard normalized dye spread rates of Hs578T cells treated with 1,2,4 and 8 μmol/L PP2 were 1.60 ± 0.08,2.00 ± 0.05,2.20 ± 0.05 and 2.70 ± 0.09,respectively (all P<0.01). Moreover,compared to control group at the same time points,the standard normalized dye spread of Hs578T cells treated with 8 μmol/L PP2 for 6,12 and 24 h were 1.4 ± 0.05,1.7 ± 0.06,and 2.2 ± 0.07,respectively (all P<0.01). Western blot showed that the expression ratios of Src kinase/β-actin of Hs578T cells treated with 1,2,4 and 8 μmol/L PP2 for 24 h were 0.93 ± 0.02,0.70 ± 0.09,0.66 ± 0.09 and 0.36 ± 0.10,which were significantly inhibited compared with control group (P<0.05 or 0.01). And the expression ratio of Src kinase/β-actin of Hs578T cells treated with 8 μmol/L PP2 for 6,12 and 24h was 0.82 ± 0.03,0.66 ± 0.08 and 0.59 ±0.09, which were all inhibited significantly compared to control group (P<0.01).

CONCLUSIONPP2 enhances the gap junction function in breast cancer Hs578T cells, which is probably related to the inhibition of Src kinase.

Breast Neoplasms ; pathology ; Cell Line, Tumor ; Female ; Gap Junctions ; drug effects ; Humans ; Pyrimidines ; administration & dosage ; pharmacology ; src-Family Kinases ; metabolism

BACKGROUNDNicotine, a major component of tobacco, is the main cause of smoking addiction. It was found that asthmatic patients who smoke were insensitive to glucocorticoid treatment. In this paper, we investigated whether nicotine could inhibit histone deacetylase 6 activity (HDAC6) and chaperone-dependent activation of the glucocorticoid receptor (GR) in A549 cells. Furthermore, the expression level of heat shock protein 90 (HSP90) was determined.

METHODSQuantitative real-time polymerase chain reaction was used to detect the levels of RNA transcription, and Western blotting was applied to analyze the levels of protein expression of HDAC6, GR, and HSP90 in A549 cells. Moreover, the effects of dexamethasone and trichostatin A were observed in A549 cells.

RESULTSA549 cell proliferation was inhibited in the presence of nicotine, and the level of RNA and protein expression of HDAC6 and GR were down-regulated.

CONCLUSIONSNicotine could inhibit HDAC6 activity and chaperone-dependent activation of GR. This might be the main reason why asthmatic patients who smoke show insensitivity to the glucocorticoid treatment.

Cell Line, Tumor ; Cell Proliferation ; drug effects ; Enzyme Activation ; drug effects ; Histone Deacetylase 6 ; Histone Deacetylases ; genetics ; metabolism ; Humans ; Nicotine ; pharmacology ; Receptors, Glucocorticoid ; genetics ; metabolism

Based on ninety three acetylcholinesterase inhibitors (AChEIs) which have the same mechanism of action but are different in structural characteristics, the pharmacophore model for acetylcholinesterase inhibitor was constructed by the CATALYST system. The optimal pharmacophore model with three hydrophobic units, a ring aromatic unit and a hydrogen-bond acceptor unit were confirmed (Weight = 3.29, RMS = 0.53, total cost-null cost = 62.75, Correl = 0.93, Config = 19.05). This pharmacophore model will act on the double active site of acetylcholinesterase and is able to predict the activity of known acetylcholinesterase inhibitors that are used for clinical treatment of Alzheimer's disease (AD), and can be further used to identify structurally diverse compounds that have higher activity treating with Alzheimer's disease (AD) by virtual screening.
Acetylcholinesterase ; chemistry ; metabolism ; Alzheimer Disease ; enzymology ; prevention & control ; Cholinesterase Inhibitors ; chemistry ; classification ; therapeutic use ; Drug Design ; Humans ; Models, Chemical ; Models, Molecular ; Molecular Structure ; Quantitative Structure-Activity Relationship ; Structure-Activity Relationship

Ras/Raf/MEK/ERK singal transduction plays an important role in cell proliferation, differentiation, apoptosis, metastasis and metabolism. This investigation focused on this signal pathway and chose farnesyl transferase (FTase) as the main target and Raf-1 kinase as the second target. A lot of compounds were selected to construct the pharmacophore models of farnesyl transferase inhibitors (FTIs) and Raf-1 kinase inhibitors by using computer-aided drug design (CADD). The pharmacophore of FTIs is constituted by a hydrogen bonding acceptor, an aromatic ring, a positive ionizable and two hydrophobic regions; the pharmacophore of Raf-1 kinase is constituted by a hydrogen donor, a hydrogen acceptor, a hydrophobic regions and an aromatic ring. There are some similarities between the two pharmacophores. After analysis of the constructions of these two pharmacophores, some new aminomethylbenzoic acid derivatives with good forecasting activity against both of FTase and Raf-1 kinase were designed with these new pharmacophore models.
Antineoplastic Agents ; chemical synthesis ; chemistry ; pharmacology ; Computer-Aided Design ; Drug Delivery Systems ; Drug Design ; Enzyme Inhibitors ; chemical synthesis ; chemistry ; Farnesyltranstransferase ; antagonists & inhibitors ; Molecular Structure ; Proto-Oncogene Proteins c-raf ; antagonists & inhibitors ; Signal Transduction

Objective To explore safety evaluation of the approaches of the percutaneous eilational tracheostomy(PDT)ane traeitional tracheotomy in the treatment of neurological patients. Methods The stuey eesign was a multicenter,prospective,raneomizee clinical trial. One huneree ane seventy-six cases with acute nerve trachea incision from Feb. 2010 to Feb. 2013 of 3 hospitals were selectee as our subject. They were raneomly eivieee into the traeitional group ane PDT group. The information inclueing operation time,the incieence of pneumothorax,subcutaneous emphysema,tracheal fistula,esophageal,trachea ane lung injury from complications such as infection were recoreee. Results The complication rate in traeitional group was 19. 51%(16 / 82),higher than that of PDT group(8. 51%(8 / 94),P = 0. 021). The surgery perioe in PDT group was(7. 5 ± 2. 3)min,shortee than that in traeitional group((41. 6 ± 5. 8)min,P = 0. 000). Conclusion The approach of percutaneous tracheostomy can quickly buile airway of neurological patients with character of simple, safe,ane it also can reeuce the incieence of respiratory complications.

Objective To explore standardized continuing vascular ultrasound training in China.Methods Data of current ultrasound practitioner education levels in the secondary and tertiary hospitals were collected.Under the direction of the Chinese Ultrasound Doctor Association (CUDA),Vascular Ultrasound Guidelines were introduced.The CUDA Steering Committee for Standardized Vascular Ultrasound Training and Vascular Ultrasound Training Centers were established.Results The questionnaires were received from 182 ultrasound departments of 57 secondary hospitals and 125 tertiary hospitals.The proportion of ultrasound practitioners who obtained a university degree was significantly higher in the ultrasound departments of tertiary hospitals than that in the secondary hospitals (75.18% vs 52.63%).The proportion of ultrasound practitioners who only received short-term ultrasound training and did not have tertiary education was significantly higher in the ultrasound departments of secondary hospitals than that in the tertiary hospitals (8.31% vs 1.03%).The directors of ultrasound departments in both secondary and tertiary hospitals believed that ultrasound practitioners should at least have a university diploma and preferably have a university degree.The CUDA and the American Registry for Diagnostic Medical Sonography (ARDMS) co-hosted two standardized vascular ultrasound training seminars and the first Registered Physician in Vascular Interpretation (RPVI) credential examination in Beijing.A well-known vascular ultrasound reference book,Introduction to Vascular Ultrasound was translated from English into Chinese and published in September 2008.The CUDA Vascular Ultrasound Guidelines were drafted by 12 vascular ultrasound experts from China,U.S.and Australia,which were widely discussed and passed by the CUDA Standing Committee.The guidelines have been published in the Chinese Journal of Ultrasonography and on the website of CUDA (www.cuda.org.cn).The CUDA has signed a publishing contract with a medical press to publish the guideline booklet.The CUDA Steering Committee for the Standardized Vascular Ultrasound Training was established which includes 24 domestic and foreign experts.The CUDA Standardized Vascular Ultrasound Training Centers were awarded to 12 hospitals,which were selected,trained and assessed by the CUDA from 35 candidates.Each center is expected to undertake one to two standardized vascular ultrasound training courses each year.Conclusion Continuing education of vascular ultrasound introduced by CUDA has been accepted positively by a portion of Chinese doctors with ultrasound major.

Objective To investigate the relationship between the expression of CENPF in NSCLC adenocarcinoma (LUAD) and the clinical prognosis of patients and its effect on the metastasis of lung adenocarcinoma cells. Methods The expression of CENPF in LUAD and its relationship with patient prognosis were analyzed by online bioinformatics. The expression of CENPF was verified by LUAD tissue microarray immunohistochemical staining. Kaplan-Meier analysis was performed to analyze the relationship between the expression of CENPF and the prognosis of patients with lung adenocarcinoma. Cox survival hazard ratio was used to analyze the factors affecting the survival of patients. Chi-square analysis was adopted to examine the relationship between CENPF expression and clinicopathological stage and grade of patients. The expression of CENPF in NCI-H2126 cells were knocked out by lentivirus, and then the proliferation, invasion, and migration abilities of the cells were detected. Changes in mRNA expression profiles after CENPF knockout were detected by RNA-seq. Bioinformatics analysis of downstream signaling pathways and the target genes of CENPF was also performed. Western blot was used to verify the target gene. Results CENPF was significantly upregulated in LUAD tumor tissue (P < 0.05) and significantly correlated with pathological stage (P=0.013). The higher expression of CENPF, the worse the prognosis of patients (P=0.01, P=0.027). After the expression was CENPF of knocked out, the cell proliferation, migration, and invasion abilities significantly reduced (P < 0.01). The expression of chemokine pathway genes in cells was enriched significantly (P < 0.001). ACKR3/CXCR7 and CDH2/N-cadherin were significantly downregulated, whereas CDH1/E-cadherin was significantly upregulated. After CENPF was knocked out, ACKR3/CXCR7 and N-cadherin were significantly downregulated, whereas E-cadherin significantly increased. Conclusion The expression of CENPF is negatively correlated with the clinical prognosis of patients with LUAD, and it promotes the occurrence of EMT by regulating the expression levels of N-cadherin and E-cadherin related to EMT through ACKR3/CXCR7.