Heat shock protein 90 protects rat mesenchymal stem cells against hypoxia and serum deprivation-induced apoptosis via the PI3K/Akt and ERK1/2 pathways

Mesenchymal stem cell(MSC)transplantation has shown a therapeutic potential to repair the ischemic and infracted myocardium,but the effects are limited by the apoptosis and loss of donor cells in host cardiac microenvironment.The aim of this study is to explore the cytoprotection of heat shock protein 90(Hsp90)against hypoxia and serum deprivation-induced apoptosis and the possible mechanisms in rat MSCs.Cell viability was determined by3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide(MTT)assay.Apoptosis was assessed by Hoechst 33258nuclear staining and flow cytometric analysis with annexin V/PI staining.The gene expression of Toll-like receptor-4(TLR-4)and V-erb-b2 erythroblastic leukemia viral oncogene homolog 2(ErbB2)was detected by real-time polymerase chain reaction(PCR).The protein levels of cleaved caspase-3,Bcl-2,Bcl-xL,Bax,total-ERK,phospho-ERK,totaI-Akt,phospho-Akt,and Hsp90 were detected by Western blot.The production of nitric oxide was measured by spectrophotometric assay.Hsp90 improves MSC viability and protects MSCs against apoptosis induced by serum deprivation and hypoxia.The protective role of Hsp90 not only elevates Bcl-2/Bax and Bcl-xL/Bax expression and attenuates cleaved caspase-3 expression via down-regulating membrane TLR-4 and ErbB2 receptors and then activating their downstream PI3K/Akt and ERK1/2 pathways,but also enhances the paracrine effect of MSCs.These findings demonstrated a novel and effective treatment strategy against MSC apoptosis in cell transplantation.     

Protective effect of hydrogen-rich saline on ischemia/reperfusion injury in rat skin flap

Objective

Skin damage induced by ischemia/reperfusion (I/R) is a multifactorial process that often occurs in plastic surgery. The mechanisms of I/R injury include hypoxia, inflammation, and oxidative damage. Hydrogen gas has been reported to alleviate cerebral I/R injury by acting as a free radical scavenger. Here, we assessed the protective effect of hydrogen-rich saline (HRS) on skin flap I/R injury.

Methods

Abdominal skin flaps of rats were elevated and ischemia was induced for 3 h; subsequently, HRS or physiological saline was administered intraperitoneally 10 min before reperfusion. On postoperative Day 5, flap survival, blood perfusion, the accumulation of reactive oxygen species (ROS), and levels of cytokines were evaluated. Histological examinations were performed to assess inflammatory cell infiltration.

Results

Skin flap survival and blood flow perfusion were improved by HRS relative to the controls. The production of malondialdehyde (MDA), an indicator of lipid peroxidation, was markedly reduced. A multiplex cytokine assay revealed that HRS reduced the elevation in the levels of inflammatory cytokines, chemokines and growth factors, with the exception of RANTES (regulated on activation, normal T-cell expressed and secreted) growth factor. HRS treatment also reduced inflammatory cell infiltration induced by I/R injury.

Conclusions

Our findings suggest that HRS mitigates I/R injury by decreasing inflammation and, therefore, has the potential for application as a therapy for improving skin flap survival.     

舒尼替尼对结肠成纤维细胞体内外抑制机制研究(英文)

研究目的:探索小分子酪氨酸激酶抑制剂舒尼替尼(sunitinib)对结肠肿瘤微环境中的肿瘤相关成纤维细胞的作用及其机制。创新要点:舒尼替尼通过抑制肿瘤间质成纤维细胞的生长,间接发挥抗肿瘤效应,为结肠癌综合治疗的提供新途径。研究方法:通过细胞周期分析和细胞增殖测定进行舒尼替尼体外抑制肿瘤细胞的研究。采用Western-blot检测磷酸化血小板衍生生长因子β受体(PDGFR-β)、蛋白激酶B(Akt)及细胞外信号调节蛋白激酶(ERK)的蛋白水平。通过注射肠腺癌细胞株SW620和结肠成纤维细胞构建的裸鼠移植瘤模型来研究舒尼替尼的体内抑瘤效果。重要结论:舒尼替尼可有效抑制结肠癌来源的原代结肠成纤维细胞生长,该抑制作用主要通过抑制血小板衍生生长因子(PDGF)信号通路得以实现。     

Methylenetetrahydrofolate reductase C677T polymorphism predicts response and time to progression to gemcitabine-based chemotherapy for advanced non-small cell lung cancer in a Chinese Han population

Objective:The aim of this study was to evaluate the association between the methylenetetrahydrofolate reductase(MTHFR) C677T excision repair cross-complementation group 1(ERCC1) genetic polymorphisms and the clinical efficacy of gemcitabine-based chemotherapy in advanced non-small cell lung cancer(NSCLC).Methods:A total of 135 chemonaive patients with unresectable advanced NSCLC were treated with gemcitabine/platinum regimens.The polymorphisms of MTHFR C677T,ERCC1 C8092A,and ERCC1 C118T were genotyped using the TaqMan methods.Results:The overall response rate was 28.9%.Patients with MTHFR CC genotype had a higher rate of objective response than patients with variant genotype(TT or CT)(41.2% versus 19.1%,P=0.01).Median time to progression(TTP) of patients with MTHFR CC genotype was longer than that of patients with variant genotype(7.6 months versus 5.0 months,P=0.003).No significant associations were obtained between ERCC1 C118T and C8092A polymorphisms and both response and survival.Conclusions:Our data suggest the value of MTHFR C677T polymorphism as a possible predictive marker of response and TTP in advanced NSCLC patients treated with gemcitabine/platinum.     

Effects of orthodontic load on the periodontium of autogenously transplanted teeth in beagle dogs

Objective:To observe the periodontal healing of autogenously transplanted teeth loaded orthodontically after autotransplantation in Beagle dogs.Methods:Forty-eight teeth were autogenously transplanted,24 of which were loaded postoperatively with orthodontic force at different time points and for different durations.Periodontal healing was evaluated by probing pocket depth(PPD),the expression of relevant proteins,and histomorphometric analyses.Results:The dental pockets of loaded and non-loaded teeth were both much deeper after the first postoperative week than before transplantation(P<0.05).Later,the PPD,which was measured after postoperative weeks 1,3,5,9 and 13,gradually became shallow.The expressions of alkaline phosphatase(ALP)and basic fibroblast growth factor(bFGF)were higher in loaded teeth than in non-loaded teeth(P<0.05),and in groups subjected to two weeks duration of loading than in other groups at the same load time point(P<0.05).For the same load duration,the expressions of ALP and bFGF in teeth loaded after postoperative week 4 were higher than those of other treatments(P<0.05).According to histomorphometric analyses,an orthodontic force on transplanted teeth applied after postoperative weeks 4 or 8 for two weeks duration should be favorable for periodontal healing.Conclusions:It is advisable to apply an appropriate magnitude of force on autotransplanted teeth,such as orthodontic force,at appropriate time points and for a suitable duration,to achieve the optimal clinical prognosis following autogenous tooth transplantation.These results may serve as a basis for subsequent studies in humans so as to make clinical improvements.     

HER-2 overexpression and survival in colorectal cancer: a meta-analysis

Objective

Numerous studies examining the relationship between human epidermal growth factor receptor 2 (HER-2) overexpression and survival in patients with colorectal cancer (CRC) have yielded controversial results. We therefore performed a meta-analysis more precisely to estimate its prognostic value.

Methods

Published studies investigating the effect of HER-2 overexpression on CRC survival were identified; the hazard ratios (HRs) and their corresponding 95% confidence intervals (95% CIs) were pooled in terms of disease-specific or overall survival.

Results

Eleven studies were included in the meta-analysis. The pooled data showed that HER-2 overexpression was negatively related to CRC survival (HR=1.10, 95% CI: 0.77–1.44). Subgroup analyses regarding test method and study quality also demonstrated little association between HER-2 overexpression and CRC survival (HR=0.89, 95% CI: 0.50–1.29; HR=0.90, 95% CI: 0.43–1.37, respectively).

Conclusions

Regardless of several limitations, our study suggested that HER-2 overexpression probably had little impact on CRC survival.     

Delivering MC3T3-E1 cells into injectable calcium phosphate cement through alginate-chitosan microcapsules for bone tissue engineering

研究目的：本研究应用海藻酸钠-壳聚糖微囊保护成骨细胞,接种到β-磷酸三钙/磷酸钙骨水泥（β-TCP/CPC）浆料中,使β-TCP/CPC骨修复材料具有一定的细胞活性,同时提高固化后材料的孔隙率和孔径,以最终实现提高β-TCP/CPC骨水泥的降解速度,加快成骨和骨修复。创新要点：本研究首次应用海藻酸钠-壳聚糖微胶囊包封成骨细胞与CPC浆料复合,复合后实现自动细胞释放,释放出的细胞具有良好的生物学活性。研究方法：（1）高压静电成囊法制备载小鼠成骨前体细胞（MC3T3-E1）的海藻酸钙和海藻酸钠-壳聚糖微胶囊;（2）微囊化MC3T3-E1细胞,进行体外培养,使用细胞计数试剂盒（CCK-8）检测细胞活性,并用钙黄绿素-AM（Calcein-AM）和碘化丙啶（PI）进行活死细胞双重染色;（3）微囊化MC3T3-E1细胞与β-TCP/CPC浆料复合培养后,激光共聚焦扫描显微镜和环境扫描电子显微镜观测细胞在材料上的释放、粘附,CCK-8法检测材料上细胞的活力,碱性磷酸酶（ALP）检测观察细胞的分化状况,茜素红染色观察释放细胞的矿化能力。重要结论：海藻酸钠-壳聚糖微胶囊可作为可注射磷酸钙骨水泥内部接种成骨细胞并实现细胞释放的良好载体,释放出的成骨细胞具有良好的生物学活性。     

Down-regulation of eIF5A-2 prevents epithelial-mesenchymal transition in non-small-cell lung cancer cells

Background

Epithelial-mesenchymal transition (EMT) is believed to be the critical process in malignant tumor invasion and metastases, and has a great influence on improving the survival rate in non-small-cell lung cancer (NSCLC) patients. Recent studies suggested that eukaryotic initiation factor 5A-2 (eIF5A-2) might serve as an adverse prognostic marker of survival. We detected eIF5A-2 in NSCLC A549 cells, and found that the invasive capability correlates with the eIF5A-2 expression.

Methods

Transforming growth factor (TGF)-β1 was used to induce EMT in A549 cells. Western blotting, immunofluorescence, wound healing assay, and transwell-matrigel invasion chambers were used to identify phenotype changes. Western blotting was also used to observe changes of the expression of eIF5A-2. We down-regulated the eIF5A-2 expression using an eIF5A-2 siRNA and identified the phenotype changes by western blotting and immunofluorescence. We tested the change of migration and invasion capabilities of A549 cells by the wound healing assay and transwell-matrigel invasion chambers.

Results

After stimulating with TGF-β1, almost all A549 cells changed to the mesenchymal phenotype and acquired more migration and invasion capabilities. These cells also had higher eIF5A-2 protein expression. Down-regulation of eIF5A-2 expression with eIF5A-2 siRNA transfection could change the cells from mesenchymal to epithelial phenotype and decrease tumor cell migration and invasive capabilities significantly.

Conclusions

The expression of eIF5A-2 was up-regulated following EMT phenotype changes in A549 cells, which correlated with enhanced tumor invasion and metastatic capabilities. Furthermore, in the A549 cell line, the process of EMT phenotype change could be reversed by eIF5A-2 siRNA, with a consequent weakening of both invasive and metastatic capabilities.     

Novel mutation c.980_983delATTA compound with c.986C>A mutation of the FRMD7 gene in a Chinese family with X-linked idiopathic congenital nystagmus

Objective

To screen mutations in FERM domain-containing protein 7 (FRMD7) gene in two Chinese families with X-linked idiopathic congenital nystagmus (XLICN).

Methods

Common ophthalmic data and peripheral blood of two Chinese XLICN families (families A and B) were collected after informed consent. Genomic DNA was prepared from the peripheral blood of members of the two families and from 100 normal controls. Mutations in the FRMD7 gene were determined by directly sequencing polymerase chain reaction (PCR) products.

Results

We identified a novel mutation c.980_983delATTA compound with c.986C>A mutation in the 11th exon of FRMD7 in family B, and a previously reported splicing mutation c.782G>C (p.R261G) in family A. The mutations were detected in patients and female carriers, while they were absent in other relatives or in the 100 normal controls.

Conclusions

Our results expand the spectrum of FRMD7 mutations in association with XLICN, and further confirm that the mutations of FRMD7 are the underlying molecular mechanism for XLICN.     

Periodontal status and associated risk factors among childbearing age women in Cixi City of China

Objective:To investigate the periodontal status and associated risk factors among women of childbearing age to increase the awareness of oral health.Methods:The study was conducted on childbearing age women in Cixi,a city in Zhejiang Province in the southeast of China.A total of 754 women participated in periodontal examination while receiving prenatal care.Data of the women were collected from the Cixi Family Planning Commission and during an interview.Clinical periodontal indices,such as bleeding on probing(BOP),probing depth(PD),and clinical attachment level(CAL) were measured during the examination.Statistical analysis on subject-based data was performed.Results:The prevalence of periodontal disease among childbearing age women in Cixi was high(84.7%).A significant association was found between the disease and educational level,pregnancy,taking oral contraceptives,stress,alcohol consumption,overweight,dental visit,and teeth brushing(P<0.05).Women who suffered periodontal disease showed deep PD,obvious BOP,and clinical attachment loss.Among this population,pregnancy was closely associated with higher BOP percentage;teeth brushing no more than once per day or brushing for less than 1 min(P<0.001) after adjusting for age and stress.Conclusions:The periodontal status of childbearing age women in Cixi needs to be improved urgently.Attention towards the periodontal health should be warranted,especially for those in special statuses and with poor awareness.     

Long-term comparison of full-bed deep anterior lamellar keratoplasty and penetrating keratoplasty in treating keratoconus

Objective

To compare postoperative outcomes of full-bed deep anterior lamellar keratoplasty (DALK) with penetrating keratoplasty (PK) in treating keratoconus.

Methods

Seventy-five eyes of 64 patients who received full-bed DALK and 52 eyes of 51 patients who received PK between June 2000 and August 2010 were included in this retrospective study. Full-bed DALK was performed using Yao’s hooking-detaching technique. PK was performed using a standard technique. Intraoperative and postoperative complications, visual acuity, rejection, graft survival, endothelial cell density, corneal sensation recovery, and re-innervation were compared between the two groups.

Results

A best correct visual acuity of 0.5 or better was achieved in 90.7% of eyes after full-bed DALK and in 92.3% of eyes after PK (P=0.75). By the fifth postoperative year, graft endothelial cell loss reached 34.6% in the PK group vs. 13.9% in the full-bed DALK group (P<0.001). There were no statistical differences in corneal sensitivity recovery or corneal re-innervation between the groups (P>0.05). Intraoperative microperforation occurred in seven out of 75 (9.3%) eyes with a temporally postoperative double anterior chamber in two eyes in the full-bed DALK group. Postoperative complications in the PK vs. the full-bed DALK groups respectively were: rejection (7.7% vs. 0%, P=0.015), high intraocular pressure (IOP) (46.2% vs. 1.3%, P<0.001), secondary glaucoma (9.6% vs. 0%, P=0.006), complicated cataract (19.2% vs. 0%, P<0.001), and wound dehiscence (9.6% vs. 0%, P=0.006).

Conclusions

Both full-bed DALK and PK can offer long-term satisfactory visual outcomes for keratoconus. Graft rejection, secondary glaucoma, complicated cataracts, and constant endothelial cell loss were observed in eyes only after PK.     

Levosimendan versus dobutamine in critically ill patients: a meta-analysis of randomized controlled trials

Objective

To evaluate the clinical efficacy of levosimendan versus dobutamine in critically ill patients requiring inotropic support.

Methods

Clinical trials were searched in PubMed, EMBASE, and the Cochrane Central Registry of Clinical Trials, as well as Web of Science. Studies were included if they compared levosimendan with dobutamine in critically ill patients requiring inotropic support, and provided at least one outcome of interest. Outcomes of interest included mortality, incidence of hypotension, supraventricular arrhythmias, and ventricular arrhythmias.

Results

Data from a total of 3 052 patients from 22 randomized controlled trials (RCTs) were included in the analysis. Overall analysis showed that the use of levosimendan was associated with a significant reduction in mortality (269 of 1 373 [19.6%] in the levosimendan group, versus 328 of 1 278 [25.7%] in the dobutamine group, risk ratio (RR)=0.81, 95% confidence interval (CI) 0.70-0.92, P for effect=0.002). Subgroup analysis indicated that the benefit from levosimendan could be found in the subpopulations of cardiac surgery, ischemic heart failure, and concomitant β-blocker therapy in comparison with dobutamine. There was no significant difference in the incidence of hypotension, supraventricular arrhythmias, or ventricular arrhythmias between the two drugs.

Conclusions

In contrast with dobutamine, levosimendan is associated with a significant improvement in mortality in critically ill patients requiring inotropic support. Patients having cardiac surgery, with ischemic heart failure, and receiving concomitant β-blocker therapy may benefit from levosimendan. More RCTs are required to address the questions about no positive outcomes in the subpopulation in a cardiology setting, and to confirm the advantages in long-term prognosis.     

萎叶提取物增强5-氟尿嘧啶对结肠癌细胞HT29和HCT116的生长抑制作用

研究目的：探讨萎叶（PB）提取物对5-氟尿嘧啶（5-FU）抑制结肠癌细胞HT29和HCT116生长的影响。研究方法：HT29和HCT116细胞分别给予PB、5-FU以及两种药物联合治疗24小时,应用等效线图法分析PB和5-FU的药效学相互作用,AnnexinV／PI染色法检测HT29和HCT116细胞的凋丁L=情况,高效液相色谱法排除PB和5-FU间任何可能的相互化学作用。重要结论：联合PB,低剂量5-FU可以在短时间内起到细胞毒作用,而单独应用PB或5-FU治疗较联合治疗可以诱导更多细胞发生凋亡。进一步采用等效线图法分析显示PB和5-Fu的联合作用在抑制结肠癌细胞HT29和HCT116的生长中分别体现出协同和拮抗作用。因此可以认为在HT29细胞中,PB使得较低剂量5-FU发挥最大抑制结肠癌细胞生长效果,然而在HCT116细胞中,PB没有显著降低5-FU的药物浓度,说明PB和5-FU的相互作用不仅仅体现在诱导细胞凋亡方面。     

Effects of zinc-substituted nano-hydroxyapatite coatings on bone integration with implant surfaces

Objective:The purpose of this study was to investigate the effects of a zinc-substituted nano-hydroxyapatite(Zn-HA) coating,applied by an electrochemical process,on implant osseointegraton in a rabbit model.Methods:A Zn-HA coating or an HA coating was deposited using an electrochemical process.Surface morphology was examined using field-emission scanning electron microscopy.The crystal structure and chemical composition of the coatings were examined using an X-ray diffractometer(XRD) and Fourier transform infrared spectroscopy(FTIR).A total of 78 implants were inserted into femurs and tibias of rabbits.After two,four,and eight weeks,femurs and tibias were retrieved and prepared for histomorphometric evaluation and removal torque(RTQ) tests.Results:Rod-like HA crystals appeared on both implant surfaces.The dimensions of the Zn-HA crystals seemed to be smaller than those of HA.XRD patterns showed that the peaks of both coatings matched well with standard HA patterns.FTIR spectra showed that both coatings consisted of HA crystals.The Zn-HA coating significantly improved the bone area within all threads after four and eight weeks(P<0.05),the bone to implant contact(BIC) at four weeks(P<0.05),and RTQ values after four and eight weeks(P<0.05).Conclusions:The study showed that an electrochemically deposited Zn-HA coating has potential for improving bone integration with an implant surface.     

A new approach to the assessment of children's mathematical competence

The report is a translation of part of a Swedish paper entitled “On Students' Achievement in Mathematics after Finishing Comprehensive School”. The intention of the investigation was to

- diagnose the retention of some basic skills in some topics in algebra and geometry,

- reveal difficult steps in the learning processes in these topics.

Starting with a complicated question, e.g. the equation \(\frac{{3x - 2}}{2} = \frac{x}{3}\) a sequence consisting of 5–15 problems were constructed. Each new problem followed by the preceding one by taking away one or two details. \(\begin{gathered} 3(3x - 2) = 2x \\ {\text{ 9}}x - 6 = 2x \\ {\text{ 7}}x - 6 = 0 \\ {\text{ 7}}x = 6 \\ \end{gathered} \) is an example of a sequence belonging to the equation above. From about 10 complicated problems (“top-items”) and their sequences, in all 130 items, 10 sub-tests were put together in such a way that the pupils who took the test were not aware of the sequences but found no connection between the problems. Many surprising results were found, e.g., that the students scored higher on 14/(x+2)=2 than on 4/x=3, that the difficulty in finding the area of a triangle depended on the position of the triangle and that the problems “Simplify a/a ², a²/a, a/a” were of quite unequal difficulty. A discussion about the students' thinking in patterns and mechanically learning ends the report.     

肝功能对意识消失时咪达唑仑血浆浓度和脑电双频谱指数的影响(英文)

研究目的:探索不同肝功能指数对镇静药咪达唑仑的敏感性以及脑电双频谱指数(BIS)的预测意识消失的影响。创新要点:明确不同肝功能状态的患者对中枢抑制药的敏感性以及BIS预测意识消失概率发生的敏感性。指导临床医生根据患者的肝功能状态合理使用中枢抑制药以及正确利用BIS预测镇定深度。研究方法:选择意识消失作为研究终点,以50%患者意识消失时的咪唑安定的浓度以及BIS值作为观察指标。重要结论:终末期肝病患者对中枢镇静药咪唑安定更敏感,该类患者在BIS预测意识消失概率发生的敏感性存在差异。     

Optimal time for mesenchymal stem cell transplantation in rats with myocardial infarction

Background:Bone marrow mesenehymal stem cell(MSC)transplantation is a promising strategy in the treatment of myocardial infarction(MI).However,the time for transplanting cells remains controversial.The aim of this study was to find an optimal time point for cell transplantation.Methods:MSCs were isolated and cultured from Sprague-Dawley(SD) rats.MI model was set up in SD rats by permanent ligation of left anterior descending coronary artery.MSCs were directly injected into the infarct berder zone at 1 h,1 week and 2 weeks after MI,respectively.Sham-operated and MI centrel groups received equal volume of phosphate buffered saline(PBS).At 4 weeks after MI,cardiac function Was assessed by echocardiography;vessel density Was analyzed on hematoxylin-eosin stained slides by light microscopy;the apoptosis of cardiomyocytes Was evaluated by terminal deoxynucleotidy1 transferase-mediated dUTP nick end-labeling(TUNEL) assay;the expressions of proteins were analyzed by Western blot.Results:MSC transplantation improved cardiac function.reduced the apoptosis of cardiomyocytes and increased vessel density.These benefits were more obvious in l-week group than in 1-h and 2-week groups.There are more obvious increases in the ratio of bc1-2/bax and the expression of vascular endothelial growth factor(VEGF)and more obvious decreases in the expression of cleaved-caspase-3 in 1-week group than those in other two groups.Conclusion:MSC transplantation was beneficial for the recovery of cardiac function.MSC transplantation at l week post-MI exerted the best effects on increases of cardiac function,anti-apoptosis and angiogenesis.     

早期经尿道前列腺隧道式电切治疗前列腺癌近距离照射治疗术后尿潴留(英文)

研究目的:探讨早期经尿道前列腺隧道式电切治疗前列腺癌近距离照射治疗术后尿潴留的适应症及其对近距离照射治疗疗效的影响。创新要点:前列腺癌近距离照射治疗已经成为75岁以上患者的首选,前列腺癌近距离照射治疗术后尿潴留发生率为1.5%~22.0%。经尿道前列腺电切常用于药物治疗无效的反复尿潴留患者,且手术时机多在近距离照射治疗术后6个月以后。经尿道前列腺隧道式电切治疗前列腺癌近距离照射治疗术后尿潴留可在6个月内进行,患者尿道症状从IV(出现尿潴留,需要导尿)下降为0级(没有症状)或I级(轻度,中度尿频,2~3次/晚),较术前改善明显,且对近距离照射治疗疗效无明显影响。研究方法:浙江大学医学院附属邵逸夫医院2009年2月至2013年7月间所有接受前列腺癌近距离照射治疗的病例共190例,其中9例(4.7%)患者术后出现反复尿潴留,且早期行经尿道前列腺隧道式电切术治疗,回顾分析其临床资料。重要结论:前列腺癌近距离照射治疗术后反复尿潴留,在严格防护下早期行经尿道前列腺隧道式电切是安全有效的,且电切后对近距离照射治疗疗效无明显影响。