Ligustrazine suppresses platelet aggregation through inhibiting the activities of calcium sensors

Abstract Ligustrazine is widely used for the treatment of cardiovascular diseases in traditional Chinese medication. It has been reported that Ligustrazine decreases the concentration of intracellular calcium ions (Ca2+); however, the underlying mechanism remains unknown. In the present study, the effect of Ligustrazine on adenosine diphosphate (ADP)-induced platelet aggregation was evaluated using a turbidimetric approach. The changes in concentration of intracellular Ca2+ stimulated by ADP was measured using fluo-4, a fluorescent Ca2+ indicator dye. The mRNA expression of stromal interaction molecule l (STIM1) and Orai1, calcium sensor, was determined using real-time PCR. In addition, the protein expression of STIM1, Orai1, and serum/glucocorticoid-regulated protein kinase 1 (SGK1) was determined using Western blot analysis. The data demonstrated that Ligustrazine significantly suppressed platelet aggregation in a dose-dependent manner and reduced the concentration of intracellular Ca2+ triggered by ADP. Our data showed that Ligustrazine treatment inhibited the expression of STIM1 and Orai1 induced by ADP at both mRNA and protein levels, and suppressed the protein expression of SGK1. Taken together, our data indicated that Ligustrazine suppressed platelet aggregation by partly inhibiting the activities of calcium sensors, thereby suggesting that Ligustrazine may be a promising candidate for the treatment of platelet aggregation.

Effects of salinomycin on proliferation and apoptosis of oral squamous cell carcinoma / 北京大学学报(医学版)

OBJECTIVE@#To investigate the effects of salinomycin on the proliferation and apoptosis of oral squamous carcinoma cells and to further understand the mechanisms of these effects.@*METHODS@#The human oral squamous carcinoma cell line CAL-27 was cultured in different concentrations of salinomycin and cisplatin. After co-culture with 0, 1, 2, 4, 8, 16 and 32 μmol/L salinomycin or 0, 1.25, 2.5, 5, 10, 20, 40 and 80 μmol/L cisplatin for 24 hours and 48 hours, the proliferation of oral squamous carcinoma cells were detected by cell counting kit-8(CCK-8) assay. After being exposed to 0, 2, 4, 8 μmol/L salinomycin and 0, 5, 10, 20 μmol/L cisplatin for 48 hours, the cell cycle of oral squamous carcinoma cells was detected by flow cytometry assay, and Western blot analysis was performed to analyze the expressions of cysteine-containing aspartate-specific proteases-3(Caspase-3), cysteine-containing aspartate-specific proteases-9(Caspase-9), poly ADP-ribose polymerase (PARP), protein kinase B (Akt) and phosphorylated protein kinase B (p-Akt) protein in oral squamous carcinoma cells.@*RESULTS@#Both salinomycin and cisplatin significantly inhibited the proliferation of oral squamous cell carcinoma CAL-27 cells in a time- and dose-dependent manner. However, compared with the first-line chemotherapeutic drug cisplatin, salinomycin showed stronger anti-proliferation activity in oral squamous carcinoma cells than cisp-latin (P < 0.001). After being exposed to 8 μmol/L salinomycin, CAL-27 cells exhibited markedly higher proportion in quiescent/ first gap phases (40.40%±1.99% vs. 64.46%±0.90%, P < 0.05), and had a significantly lower proportion in synthesis phases and second gap / mitosis phases (24.32%±2.30% vs. 18.73%±0.61%, P < 0.05; 35.01%±1.24% vs. 16.54%±1.31%, P < 0.05) compared with the dimethyl sulfoxide control group; moreover cisplatin didn't show cell-cycle specific effect on CAL-27. Western blot proved that salinomycin could up-regulate the expressions of Caspase-3 and Caspase-9 protein in oral squamous cell carcinoma CAL-27 cells (P < 0.05). At the same time, the levels of PARP, Akt and p-Akt protein were down-regulated (P < 0.05).@*CONCLUSION@#Compared with cisplatin, salinomycin has a better inhibitory effect on the proliferation of oral squamous carcinoma cells and blocks the cell cycle process at the quiescent / first gap phase. At the same time, salinomycin could trigger apoptosis of oral squamous carcinoma cells and the mechanism is associated with the Akt/p-Akt signaling pathway.

Acute- and Long-term Outcomes of Rotational Atherectomy followed by Cutting Balloon versus Plain Balloon before Drug-Eluting Stent Implantation for Calcified Coronary Lesions / 中华医学杂志(英文版)

Background@#Coronary calcification is a major determinant of stent underexpansion and subsequent adverse events. This study aimed to evaluate the acute- and long-term outcomes of rotational atherectomy (RA) followed by cutting balloon (CB) versus plain balloon before drug-eluting stent implantation for calcified coronary lesions.@*Methods@#From June April 2013 to March 2016, a total of 127 patients with moderately or severely calcified coronary lesions were treated with RA. Patients were divided into two groups according to the balloon type after RA: RA+CB group (n = 75) and RA+plain balloon group (n = 52). Minimal lumen diameter and acute lumen gain were analyzed by quantitative coronary angiography. In-hospital and long-term (>1 year) outcomes were recorded. Multivariate Cox regression analysis was performed to determine the independent predictors of in-stent restenosis.@*Results@#The mean age of the patients was 65.5 years, and 76.4% were men. Total lesion length and minimal lumen diameter at baseline were similar in the two groups. After RA and balloon dilation, the lumen diameter was significantly larger in the RA+CB group than in the RA+plain balloon group (1.57 ± 0.46 mm vs. 1.10 ± 0.40 mm, t = 4.123, P 1 year) in-stent restenosis (hazard ratio: 0.136, 95% confidence interval: 0.020-0.936, P = 0.043).@*Conclusions@#In patients with moderately or severely calcified lesions, a strategy of RA followed by CB before stent implantation can increase lumen diameter and acute lumen gain. This strategy is safe with lower risk of long-term in-stent restenosis.

In vivo study of 3D printed porous tantalum implant on osseointegration / 华西口腔医学杂志

<p><b>OBJECTIVE</b>This work aims to investigate the effect of porous tantalum and porous titanium on osseointegration.</p><p><b>METHODS</b>Two kinds of porous materials with same microporous parameters, namely, porous tantalum and porous titanium, were fabricated by computer-aided design (CAD) modeling and 3D printing technology. A defect model was established in 24 New Zealand white rabbits in the bilateral femoral lateral malleolus at the left and right side of each animal. Then, animals were randomly divided into two groups, and bone defects were repaired by porous tantalum and porous titanium (experimental and control groups, respectively). Animals were sacrificed at two, four, and eight weeks after implantation. Gross observation and methylene blue-acid fuchsin staining were used to observe osseointegration of the implant and bone interface, and the osseointegration strength of implant bone interface was tested by push-out test.</p><p><b>RESULTS</b>At two, four, and eight weeks after operation, the new bone tissue in the two groups increased gradually, and new bone trabecula appeared and grew into the pores of the materials. No significant difference (P>0.05) in osteogenesis and the strength of implant bone tissue interface between the two groups was observed.</p><p><b>CONCLUSIONS</b>3D
printed porous tantalum implants, which exhibit comparable osseointegration capabilities to porous titanium implants, can form an early biological combination with bone tissue.</p>

SHMT1 C1420T polymorphism contributes to the risk of non-Hodgkin lymphoma: evidence from 7309 patients / 癌症

<p><b>BACKGROUND</b>Serine hydroxymethyltransferase 1 (SHMT1) is a key enzyme in the folate metabolic pathway that plays an important role in biosynthesis by providing one carbon unit. SHMT1 C1420T may lead to the abnormal biosynthesis involved in DNA synthesis and methylation, and it may eventually increase cancer susceptibility. Many epidemiologic studies have explored the association between C1420T polymorphism and the risk of non-Hodgkin lymphoma (NHL), but the results have been contradictory. Therefore, we performed this meta-analysis to evaluate the relationship.</p><p><b>METHODS</b>The meta-analyses were conducted to evaluate the effect of SHMT1 C1420T polymorphism on NHL risk. Odds ratios (ORs) and 95% confidence intervals (CIs) were calculated to measure the strength of the association.</p><p><b>RESULTS</b>Eight studies encompassing 3232 cases and 4077 controls were included. A statistically significant association was found between SHMT1 C1420T polymorphism and NHL risk under the allelic comparison (T vs. C: OR = 1.09, 95% CI 1.01-1.17); a borderline association was found between SHMT1 C1420T polymorphism and NHL risk under the homozygote model (TT vs. CC: OR = 1.18, 95% CI 1.00-1.39) and the dominant model (CT+TT vs. CC: OR = 1.10, 95% CI 1.00-1.21).</p><p><b>CONCLUSION</b>SHMT1 C1420T polymorphism may be associated with NHL risk, which needs to be validated in large, prospective studies.</p>

Effects of basic fibroblast growth factor via coronary venous retroperfusion on bone marrow mesenchymal stem cell differentiation in vivo / 中国组织工程研究

BACKGROUND:In vitro studies have demonstrated that basic fibroblast growth factor (bFGF) promote the differentiation of bone marrow mesenchymal stem cells (BMSCs) into cardiomyocyte-like cells. However, it is unclear whether coronary venous retroperfusion of bFGF stimulates BMSCs differentiation in vivo. OBJECTIVE:To evaluate the effects of coronary venous retroperfusion of bFGF on BMSCs differentiation in vivo. METHODS:BMSCs from 12 dogs were isolated by density gradient centrifugation and expanded in vitro. These cells were transfected by enhanced green fluorescence protein (EGFP) lentiviral vector and the transfection efficiency was analyzed. Acute myocardial infarction was induced by ligation of left anterior descending coronary artery. After 1 week, 10 survival animals were randomized to BMSCs group (n=5) and bFGF+BMSCs group (n=5). bFGF-and EGFP-positive BMSCs were reversely infused via coronary vein using over-the-wire bal oon catheter. One week after infusion, the number of EGFP-positive cells co-staining factor VIII and troponin I was compared between the two groups by immunofluorescence method. RESULTS AND CONCLUSION:BMSCs were successful y transfected by EGFP and the transfection efficiency was 85%. Immunofluorescence showed that EGFP-positive BMSCs were observed in 23.5%of slides. There were more EGFP-positive cells co-staining VIII and troponin I in the bFGF+BMSCs group than in the BMSCs group (P<0.05). Thus, the coronary venous retroperfusion of bFGF enhances the differentiation of BMSCs into vascular endothelial cells and cardiomyocytes. Combined delivery of bFGF and BMSCs can exert a synergistic effect to promote cardiac repair.

Cytocompatibility of two porous bioactive glass-ceramic in vitro / 华西口腔医学杂志

<p><b>OBJECTIVE</b>To compare the cytocompatibility of two kinds porous bioactive glass-ceramic made by same raw materials.</p><p><b>METHODS</b>Apatite/wollastonite bioactive glass-ceramic (4006) were prepared by sol-gel method, and bioactive glass (45S5) were prepared by melting method. Bone marrow stromal cells (BMSCs) were cultivated, differentiated and proliferated into osteoblasts, from a rabbit's marrow in the differentiatiofn culture medium with active function. The viability of BMSCs cultivated with extraction of these two kinds of biomaterial, which could represent the cytotoxicity effect of 4006 and 45S5 against BMSCs, was evaluated by the MTp assay. BMSCs were seeded and cocultivated with two kinds of biomaterial scaffolds respectively in vitro. The proliferation and biological properties of cells adhered to scaffolds were observed by inverted phase contrast microscope, scanning electron microscope (SEM), and environmental scanning electron microscope (ESEM), and a suitable cell amount for seeding on the scaffold was searched.</p><p><b>RESULTS</b>There was no difference on the viability of BMSCs only cultured for one day by complete extract of 4006 and culture medium (P>0.05), but there was significant difference between them when the cells had been cultured for 3 days(P<0.01). The extract of 45S5 had significantly higher cytotoxicity than extract of culture medium (P<0.01). The BMSCs adhered, spread, and proliferated throughout the pores of the scaffold 4006, and the amount of cells adhered to 4006 was more than to 45S5. The adhered cells to 4006 increased with the rising amount of cells seeded. And 2 x 10(7) cells.mL-1 suspension resulted inthe highest cell adherence during the comparative cells adherence test.</p><p><b>CONCLUSION</b>Apatite/woolastonite bioac tive glass-ceramic has good bioactivity and cytocompatibility. Therefore, it may have the potential to be a new cell vehicle for bone tissue engineering. And the suitable seeding cell amount of apatite/wollastonite bioactive glass-ceramic should be 2x10(7) cells.mL-1 or even more than that.</p>

Trastuzumab in combination with chemotherapy versus chemotherapy alone for first-line treatment of HER2-positive advanced gastric or gastroesophageal junction cancer: a Phase III, multi-center, randomized controlled trial, Chinese subreport / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To evaluate the efficacy and safety of trastuzumab in combination with chemotherapy versus chemotherapy alone in the first-line treatment of HER-2-positive advanced gastric or gastro-oesophageal junction cancer.</p><p><b>METHODS</b>Fifteen Chinese research centers are involved in the BO18255 (ToGA) study. Patients with gastric or gastro-oesophageal junction cancer were eligible for inclusion if their tumor showed overexpression of HER-2 protein by immunohistochemistry +++ or FISH-positive. Patients were randomly assigned in a 1:1 ratio to receive a chemotherapy regimen consisting of capecitabine or 5-FU plus cisplatin or chemotherapy in combination with intravenous trastuzumab. The primary endpoint was overall survival.</p><p><b>RESULTS</b>Eighty-five Chinese patients were enrolled in this study, of whom 84 were included in the primary analysis: trastuzumab plus chemotherapy (FP/H) (n = 36) and chemotherapy alone (FP)(n = 48). The median follow-up was 15.2 months in the FP/H group and 14.2 months in the FP group. The median survival time was 12.6 months in the FP/H group compared with 9.7 months in the FP group [hazard ratio 0.72, 95%CI (0.40; 1.29)]. Grade 3/4 adverse events were higher in the FP/H(63.9%)than FP (47.9%) groups, including neutropenia, vomiting and nausea. Two mild cardiac adverse events occurred in the FP/H group. Severe adverse events occurred in 3 cases of both two groups, respectively.</p><p><b>CONCLUSIONS</b>Addition of trastuzumab to chemotherapy is well tolerated and shows improved survival in Chinese patients with advanced gastric or gastro-oesophageal junction cancer. These results are consistent with the results of ToGA whole population trial. Trastuzumab in combination with chemotherapy can be considered as a new option for patients with HER-2-positive advanced gastric or gastro-oesophageal junction cancer.</p>

In vivo concentration gradient of basic fibroblast growth factor after coronary venous retrograde perfusion / 中国组织工程研究

10.3969/j.issn.2095-4344.2013.24.015

Relationship between polymorphisms of related genes in the folic acid metabolic pathway and the survival of gastric cancer patients treated with capecitabine combined with paclitaxel / 肿瘤

Objective: To investigate the association of polymorphisms of related genes in folic acid metabolic pathway and the tumor candidate genes with the survival of gastric cancer patients treated with capecitabine combined with paclitaxel. Methods: Ninety-three patients with pathological diagnosis of gastric cancer were treated with capecitabine plus paclitaxel-based chemotherapy. The gene polymorphisms of tumor necrosis factor ( TNF) A308G (rs1800629), methylenetetrahydrofolate reductase ( MTHFR) C677T (rs1801131) and A1298C (rs1801133), methionine synthase ( MTR) A2756G (rs1805087) and methionine synthase reductase ( MTRR) A66G (rs1801394) were detected by TaqMan-MGB probe typing method. The median survival time (MST) of patients with different genotypes was compared, and the effects of various factors on the prognosis were evaluated. Results: The median follow-up period was 29.6 months. The MST of 93 patients was 34.93 months. The MST of patients with MTHFR 1298CA and 1298CC were 47.50 and 22.91 months, respectively. The log-rank test revealed that the polymorphism of MTHFR 1298C/A had marginally significant correlation with the survival of patients (χ2=3.447, P=0.062), and other gene polymorphisms were not associated with the survivals. The gender, drinking and operation were the major prognostic factors for gastric cancer patients receiving chemotherapy. Conclusion: Detection of MTHFR 1298CA polymorphism may predict the survival of gastric cancer patients receiving chemotherapy of capecitabine combined with paclitaxel.

Differentiation of human periodontal ligament stem cells into neuron-like cells in vitro / 华西口腔医学杂志

<p><b>OBJECTIVE</b>To isolate and purify the human periodontal ligament stem cells (PDLSC) and investigate the differentiation potentials of PDLSC into neuron-like cells in vitro.</p><p><b>METHODS</b>PDLSC were isolated and cultivated. PDLSC of passage 2 was plated at a density of 5 x 10(3) per mL. At 80% confluence, the PDLSC were preinduced for 24 hours, and were subsequently replaced with an inducing medium containing certain concentration of 13-mercaptoethanal (beta-ME). After 6 hours of induction, the results were evaluated by morphological observation, immunocytochemical staining for neuron specific enolase (NSE), neurofilament (NF) and glial fibrillary acid protein (GFAP) expression and RT-PCR for NSE, NF, GFAP mRNA. Meanwhile, the uninduced PDLSC were used as a negative control.</p><p><b>RESULTS</b>PDLSC could be differentiate into cells with typical neuronal morphology. Immunohisto-chemistry and RT-PCR confirmed that the induced cells expressed NSE and NF, two marked enzymes of neuron cell.</p><p><b>CONCLUSION</b>PDLSC can be induced into neuron-like cells in vitro. PDLSC have the capability of multilineage differentiations.</p>

Relationship between serum VEGF level and clinical feature in children and adolescent with lymphoid malignancy / 白血病·淋巴瘤

Objective To investigate the relationship between serum-VEGF(sVEGF)and clinical features in children and adolescent patients with non-Hodgkin lymphoma(NHL) and acute lymphoblagtic leukemia(ALL).Methods The sVEGF in 101 of pretreated NHL and ALL patients were detected by enzymelinked inununosorbent assay(ELISA).The sVEGF prior and post-treatment were compared in 61 patients who achieved complete remission(CR).Results The median sVEGF was 567.70 ng/L in 81 prior-treated NHL patients.It was significantly higher than that in normal controls(P＜0.001).The median sVEGF wag 253.90 ng/L in 49 patients with CR,which was significantly different compared to pretherapeutic level(P＜0.001),whereag no statistical difference was observed compared to the normal controls. No relationships were found between sVEGF and clinical indexes such as clinical stage,Bsymptoms,gender,performance status(PS)score,bulk and serum lactate dehydrogenage (LDH)et al in untreated NHL patients.The median sVEGF was 198.60 ng/L in 20 untreated ALL patients.which wag no statistically different in comparison with that of normal controls.And the median sVEGF wag 181.73 ng/L in 12 of the CR ALL patients.which wag not statistically different in comparison with that in prior-treatment group or normal controls.Conclusion This study showed that the sVEGF in untreated children and adoleseent patients with NHL were higher than that of normal controls.The high sVEGF dmpped after achieving CR.There was no relationship between the level of sVEGF and clinical characteristics in the NHL patients.The sVEGF level in untreated ALL patients wag not difierent compared to that of the normal controls.and there was no change for sVEGF after chemotherapy in ALL patients.

Oxaliplatin Induced Neurotoxicity: Characteristic and Therapy / 上海第二医科大学学报

Oxaliplatin, the third generation of the platinum based chemotherapy agent, is effective in the treatment of multiple solid tumors and is also quite safe. However, there is a high incidence of peripheral neurotoxiciy, which is dose-limiting. Oxaliplatin induces two distinct forms of neurotoxicity: an acute syndrome that is triggered or aggravated by exposure to cold, and chronic cumulative sensory neurotoxicity which in nature resembles characteristics of cisplatin associated neurotoxicity. In this article, the clinical manifestations, electrophysiologic abnormalities, mechanism of neurotoxicity and therapy are reviewed.

Association between functional genetic polymorphisms of IL-1B and IL-1RN and susceptibility to gastric cancers / 上海第二医科大学学报

Objective To study the association between functional genetic polymorphisms of IL-1B(T-31C,C-511T),IL-1RN and the susceptibility to gastric cancers. Methods A case-control study was conducted in 180 gastric cancer cases and 308 age-and sex-matched cancer-free controls.Genotypes were detected by PCR-restriction fragment length polymorphism(PCR-RFLP) assays,and association between genotypes,environmental factors and risk of gastric cancers were determined. Results IL-1B T-31C was in strong linkage disequilibrium with IL-1B C-511T(D'=0.862,R2= 0.721,P=0.000).Multivariate logistic regression analysis revealed that the variant genotypes of IL-1B T-31C and C-511T were not significantly associated with risks for gastric cancers(adjusted OR,0.95 and 95% CI,0.62-1.47 for IL-1B T-31C;and adjusted OR,0.85 and 95% CI,0.55-1.31 for IL-1B C-511T).The variant genotypes(1/2,2/2) in IL-1RN were associated with a non-significantly increased risks for gastric cancers(adjusted OR,1.32 and 95% CI,0.71-2.36) in all subjects and with a significantly increased risks for gastric cancers in subjects with H.pylori infection(adjusted OR,2.03 and 95%CI,1.02-4.80).Conclusion The functional genetic polymorphisms of IL-1RN may contribute to the risks of gastric cancers in high-risk population,particularly in those with H.pylori infection.