Update on pediatric corneal diseases and keratoplasty

Managing pediatric corneal disorders is challenging as the prognosis of pediatric keratoplasty depends on several factors. Advancements in the genetic basis of congenital corneal diseases and investigations in congenital corneal conditions provide a better understanding of pediatric corneal conditions. Surgeons performing keratoplasty in children now have a choice of various techniques. Evolving surgical techniques of anterior lamellar and endothelial keratoplasties have expanded the management interventions in these pediatric corneal morbidity conditions; however, considerable concerns still exist in association with corneal transplantation in infants and children. Outcomes in pediatric keratoplasty depend upon the preoperative indications, the timing of surgical intervention, intraoperative and postoperative factors including the patient/care givers’ compliance. Factors such as low scleral rigidity, higher rate of graft failure, need for frequent examinations under anesthesia, and difficulty in optimal visual acuity assessment still remain a considerable challenge in pediatric scenarios. In children, deprivation amblyopia as a result of the corneal opacification can adversely affect visual development, causing dense amblyopia. Outcomes to surgical interventions for management of corneal opacification in children are further compromised by the preexisting amblyopia apart from the concerns of refractive outcome of the graft. Graft rejection, graft infection, amblyopia, and glaucoma continue to be serious concerns. In recent years both anterior and posterior lamellar keratoplasty techniques are being increasingly performed in pediatric eyes, which offer advantages in the form of lower risk of graft rejection. The timing of surgery, careful case selection, cautious intraoperative approach, and optimal postoperative management can improve the anatomical and functional outcome in difficult cases.     

Utility of Differential White Cell Count and Cell Population Data for Ruling Out COVID-19 Infection in Patients With Community-Acquired Pneumonia

IntroductionThe main aim of this study was to assess the utility of differential white cell count and cell population data (CPD) for the detection of COVID-19 in patients admitted for community-acquired pneumonia (CAP) of different etiologies.MethodsThis was a multicenter, observational, prospective study of adults aged ≥18 years admitted to three teaching hospitals in Spain from November 2019 to November 2021 with a diagnosis of CAP. At baseline, a Sysmex XN-20 analyzer was used to obtain detailed information related to the activation status and functional activity of white cells.ResultsThe sample was split into derivation and validation cohorts of 1065 and 717 patients, respectively. In the derivation cohort, COVID-19 was confirmed in 791 patients and ruled out in 274 patients, with mean ages of 62.13 (14.37) and 65.42 (16.62) years, respectively (p < 0.001). There were significant differences in all CPD parameters except MO-Y. The multivariate prediction model showed that lower NE-X, NE-WY, LY-Z, LY-WY, MO-WX, MO-WY, and MO-Z values and neutrophil-to-lymphocyte ratio were related to COVID-19 etiology with an AUC of 0.819 (0.790, 0.846). No significant differences were found comparing this model to another including biomarkers (p = 0.18).ConclusionsAbnormalities in white blood cell morphology based on a few cell population data values as well as NLR were able to accurately identify COVID-19 etiology. Moreover, systemic inflammation biomarkers currently used were unable to improve the predictive ability. We conclude that new peripheral blood biomarkers can help determine the etiology of CAP fast and inexpensively.     

异基因造血干细胞移植患者生活质量变化轨迹及影响因素研究

目的 探讨异基因造血干细胞移植患者3年内的生活质量变化轨迹及影响因素。方法 采用便利抽样法,选取2016年7月—2017年10月于苏州市某三级甲等医院行异基因造血干细胞移植的患者作为调查对象,在患者移植前（入仓前1~2 d）、移植后1个月、3个月、半年、1年、1年半及3年时采用一般资料调查表及癌症治疗功能评价-骨髓移植测评量表进行调查。 结果 异基因造血干细胞移植患者的癌症治疗功能评价-骨髓移植测评量表总分在移植后1个月时降至最低,此后呈逐渐上升趋势,半年时基本恢复至移植前水平,3年时总分显著高于移植前,差异具有统计学意义（P<0.05）。生理健康、功能健全及干细胞移植3个维度得分与癌症治疗功能评价-骨髓移植测评量表总分变化轨迹一致,呈波动上升型;情绪稳定维度在移植前得分最低,随移植时间延长逐渐上升;社交/家庭健全维度移植前得分与移植后各时间点得分比较,差异无统计学意义（P>0.05）。多元线性回归分析结果显示,影响患者移植后3年时生活质量的主要因素为居住地（t=3.175,P=0.002）、家庭人均月收入（t=3.320,P=0.001）、移植相关并发症（t=-6.955,P<0.001）及患者是否回归工作/学习（t=2.706,P=0.008）。 结论 异基因造血干细胞移植患者总体生活质量呈波动上升趋势,癌症治疗功能评价-骨髓移植测评量表5个维度间的恢复时间及变化轨迹存在差异。至移植后3年时,居住地为农村、家庭人均月收入较低、患有移植相关并发症及尚未回归工作/学习的患者生活质量更差。护理人员可根据患者生活质量变化轨迹动态调整护理措施,为其提供更为精准的延续性护理。     

Nidogen 1 regulates proliferation and migration/invasion in murine claudin-low mammary tumor cells

Nidogen 1 (NID1) is a glycoprotein found in basement membranes involved in cross-linking collagen IV and laminin. The role of NID in breast cancer has only been evaluated in a small number of studies and the findings of these studies have been inconsistent. Our previous work revealed that highly tumorigenic murine mammary tumor cells express high levels of Nid1 while weakly tumorigenic mammary tumor cells express low levels of Nid1. To investigate Nid1, two stable knockdown lines were created, and Nid1 knockdown was confirmed at both the mRNA and protein level. Nid1 knockdown significantly reduced cell proliferation and migration/invasion and these reductions in proliferation and migration/invasion could be rescued by conditioned media containing NID1 protein. The reduced migration/invasion observed in the Nid1 knockdown cells was not associated with significant alterations in the epithelial gene Cdh1 or the mesenchymal genes Snai1, Snai2, Twist1, Twist2, Zeb1 and Zeb2. Therefore, suppression of Nid1 expression reduces proliferation and migration/invasion in claudin-low murine mammary tumor cells.     

葫芦素抗肿瘤作用研究进展

癌症是全球主要的公共卫生问题，中国国家癌症中心的统计数据也表明癌症以逐年提升的发病率和死亡率成为威胁人类健康的重大疾病。癌症的发生发展机制复杂，涉及多阶段、多个基因及多条信号通路的异常，常规的放化疗及新兴的靶向治疗方法是肿瘤治疗的主要方式，但由于其毒副作用和耐药性的产生严重影响了患者的生活质量和持续有效的治疗效果，因此寻找安全有效的抗癌药物是全球关注的焦点。抗肿瘤植物药紫杉醇类、长春碱类、鬼臼素类、人参皂苷和多糖等的研发与应用给肿瘤治疗带来了新的希望。葫芦素是一种来源于中药葫芦科植物的高度氧化的四环三萜类化合物，药理作用广泛且机制复杂，在葫芦素家族中，目前针对葫芦素B，D，E，I抗肿瘤作用研究最多，已有大量研究证实，葫芦素B，D，E，I在治疗消化系统、呼吸系统、生殖系统、血液系统、泌尿系统等肿瘤疾病中发挥了重要作用，能够显著抑制肿瘤细胞增殖，而且在阻滞细胞周期、诱导细胞凋亡和自噬性死亡、抑制细胞迁移和侵袭、抑制肿瘤血管生成、调节活性氧的水平、调节免疫等方面作用显著，有望开发成为抗癌新药。基于当前国内外针对葫芦素抗多种肿瘤的作用及机制研究成果，笔者对葫芦素抗肿瘤作用研究进展进行了全方面综述，以期为葫芦素类抗肿瘤药物新药研发提供参考。     

Molecular mechanisms and cellular events involved in the neuroprotective actions of estradiol. Analysis of sex differences

Estradiol, either from peripheral or central origin, activates multiple molecular neuroprotective and neuroreparative responses that, being mediated by estrogen receptors or by estrogen receptor independent mechanisms, are initiated at the membrane, the cytoplasm or the cell nucleus of neural cells. Estrogen-dependent signaling regulates a variety of cellular events, such as intracellular Ca²⁺ levels, mitochondrial respiratory capacity, ATP production, mitochondrial membrane potential, autophagy and apoptosis. In turn, these molecular and cellular actions of estradiol are integrated by neurons and non-neuronal cells to generate different tissue protective responses, decreasing blood-brain barrier permeability, oxidative stress, neuroinflammation and excitotoxicity and promoting synaptic plasticity, axonal growth, neurogenesis, remyelination and neuroregeneration. Recent findings indicate that the neuroprotective and neuroreparative actions of estradiol are different in males and females and further research is necessary to fully elucidate the causes for this sex difference.     

基于Akt/ERK探讨丹蒌片对主动脉平滑肌细胞增殖、迁移和凋亡的影响＊

目的 观察丹蒌片对同型半胱氨酸（homocysteine，Hcy）诱导的兔主动脉血管平滑肌细胞（rabbit aortic smooth muscle cell，RASMC）增殖、迁移和凋亡的影响，并探讨其与Akt及ERK信号通路的关系。方法 培养RASMC，建立Hcy诱导RASMC增殖模型，实验分为空白组、Hcy组、丹蒌片组、瑞舒伐他汀组（即可定组）、丹蒌片联合瑞舒伐他汀组（即混合组），后三组分别给于丹蒌片、瑞舒伐他汀及丹蒌片联合瑞舒伐他汀含药血清。作用24 h后，Cell Titer-Glo法测细胞增殖，Transwell法测细胞迁移，流式细胞仪Annexin V/PI双染法测细胞凋亡率，蛋白印迹法（Western blot）测信号通路相关蛋白Akt和ERK总蛋白及磷酸化蛋白表达量。结果 ①与空白组相比，Hcy组细胞增殖和迁移显著增加（P < 0.01），凋亡率显著降低（P < 0.01）；与Hcy组相比，三种含药血清组细胞增殖和迁移明显减少（P < 0.01），凋亡率明显增加（P < 0.01）；其中混合组增殖和迁移较丹蒌片组和可定组明显降低（P < 0.01），凋亡率更高（P < 0.01）；与可定组比，丹蒌片组增殖略高，但差异均无统计学意义（P > 0.05），丹蒌片组细胞迁移数明显多于可定组（P < 0.05），凋亡率明显低于可定组（P < 0.05）。②各组Akt及ERK总蛋白表达水平无明显变化（P > 0.05）；与空白组比较，Hcy组p-Akt及p-ERK蛋白表达量均显著上调（P < 0.01）；与Hcy组相比，三种含药血清组p-Akt及p-ERK蛋白表达量明显下调（P < 0.01）；其中混合组的p-Akt及p-ERK蛋白表达较丹蒌片组和可定组进一步下降（P < 0.01）；与可定组比，丹蒌片组p-Akt及p-ERK蛋白表达量增加，但差异无统计学意义（P > 0.05）。结论 丹蒌片可拮抗Hcy诱导的兔动脉血管平滑肌细胞增殖和迁移，促进其凋亡，其作用可能与上调p-Akt及p-ERK信号通路蛋白有关。     

Cytotoxic and Antimetastatic Activity of Hesperetin and Doxorubicin Combination Toward Her2 Expressing Breast Cancer Cells

Objective: This study aimed to explore Hesperetin (Hst) potency as a co-chemotherapeutics agent combined with Doxorubicin (Dox), particularly cytotoxic and antimetastasis effects toward MCF-7/HER2 cells. Methods: The cytotoxic effects were measured under MTT assay. The flowcytometry analysis was used to examine the cell cycle modulation and apoptosis evidence, while the effect of migration was assayed by scratch wound healing assay. Western blotting and gelatin zymography were carried out to examine the expression level of proteins, HER2, and Rac1. Results: Under MTT assay, Hst and Dox exhibited to decrease cell viability in a dose-dependent manner with the IC50 value of 377 and 0,8 µM, respectively. The combination of Hst and Dox at the respective doses of 95 and 0,2 µM showed a synergistic effect with the combination index of 0,63. Flow cytometry analysis of Hst-Dox revealed that those compounds caused cell cycle arrest at the G2/M phase and induced apoptosis. Hst also decreased HER2 and Rac1 expression, as shown by western blot. Hst inhibited lamellipodia formation and cell migration, as indicated by microscopic observation and wound healing scratch assay. The antimetastatic activity of Hst was associated with the reduction of Rac1 and MMP9 expression as measured by gelatine zymography assay. Conclusion: These results indicated that the combination of Hst and Dox-induced cell cycle arrest, apoptosis, decreased HER2, Rac1, MMP9 expression, and cell migration. Thus, Hst may have the potential to be developed as a co-chemotherapeutic agent combined with doxorubicin toward HER2 overexpressing breast cancer cells.     

益气温阳活血利水方对AngⅡ诱导H9c2心肌细胞线粒体凋亡途径的影响

目的观察益气温阳活血利水方含药血清对AngII诱导H9c2心肌细胞线粒体凋亡途径相关Bax、Bcl-2、Caspase-9基因及蛋白的表达变化,探讨益气温阳活血利水方对AngII诱导H9c2心肌细胞凋亡的保护作用的机制。方法AngII诱导H9c2心肌细胞凋亡模型,采用血清药理学方法制备益气温阳活血利水方含药血清并进行干预,使用qRT-PCR、Western Blot、免疫组化方法检测Caspase-9 mRNA和蛋白及Bax、Bcl-2蛋白表达变化。结果益气温阳活血利水方含药血清可提高Bcl-2蛋白表达,降低Bax蛋白表达,下调Caspase-9基因及蛋白的表达水平,且含药血清高剂量组改善作用最好。结论益气温阳活血利水方含药血清对AngII诱导H9c2心肌细胞凋亡的保护作用是通过调节线粒体凋亡途径相关Bcl-2、Bax基因蛋白的表达,从而抑制Caspase-9的表达实现的。     

Cytotoxic Effects of Betel Quid and Areca Nut Aqueous Extracts on Mouse Fibroblast,Human Mouth-Ordinary-Epithelium 1 and Human Oral Squamous Cell Carcinoma Cell Lines

Background: Betel quid chewing is more common among the older generation in rural areas of Malaysia. Oral cancer in Asia has been associated with the habit of chewing betel quid and areca nut. Objective:  This study aims to investigate the cytotoxic effects of betel quid and areca nut extracts on the fibroblast (L929), mouth-ordinary-epithelium 1 (MOE1) and oral squamous cell carcinoma (HSC-2) cell lines. Methods: L929, MOE1 and HSC-2 cells were treated with 0.1, 0.2 and 0.4 g/ml of betel quid and areca nut extracts for 24, 48 and 72 h. MTT assay was performed to assess the cell viability. Results: Both extracts, regardless of concentration, significantly reduced the cell viability of L929 compared with the control (P<0.05). Cell viability of MOE1 was significantly enhanced by all betel quid concentrations compared with the control (P<0.05). By contrast, 0.4 g/ml of areca nut extract significantly reduced the cell viability of MOE1 at 48 and 72 h of incubation. Cell viability of HSC-2 was significantly lowered by all areca nut extracts, but 0.4 g/ml of betel quid significantly increased the cell viability of HSC-2 (P<0.05). Conclusion: Areca nut extract is cytotoxic to L929 and HSC-2, whereas the lower concentrations of areca nut extract significantly increased the cell viability of MOE1 compared to the higher concentration and control group. Although betel quid extract is cytotoxic to L929, the same effect is not observed in MOE1 and HSC-2 cell lines. Further investigations are needed to clarify the mechanism of action.