Crispr-associated (Cas) effectors delivery via microfluidic cell-deformation chip

Identifying new and even more precise technologies for modifying and manipulating selectively specific genes has provided a powerful tool for characterizing gene functions in basic research and potential therapeutics for genome regulation. The rapid development of nuclease-based techniques such as CRISPR/Cas systems has revolutionized new genome engineering and medicine possibilities. Additionally, the appropriate delivery procedures regarding CRISPR/Cas systems are critical, and a large number of previous reviews have focused on the CRISPR/Cas9�12 and 13 delivery methods. Still, despite all efforts, the in vivo delivery of the CAS gene systems remains challenging. The transfection of CRISPR components can often be inefficient when applying conventional delivery tools including viral elements and chemical vectors because of the restricted packaging size and incompetency of some cell types. Therefore, physical methods such as microfluidic systems are more applicable for in vitro delivery. This review focuses on the recent advancements of microfluidic systems to deliver CRISPR/Cas systems in clinical and therapy investigations. © 2021 by the authors. Licensee MDPI, Basel, Switzerland

Hesperidin improves the follicular development in 3D culture of isolated preantral ovarian follicles of mice

In vitro follicular culture systems provide optimal culture models for research about the physiology of the ovary and support the clinical practices to achieve competent mature oocytes for in vitro fertilization. In vitro maturation of preantral follicles makes it possible to study the effects of therapeutic agents on various conditions or disorders of the ovary. Nowadays, preventive bioflavonoids against cancer, hypercholesterolemia, fatty liver, or a variety of toxic agents are in focus. The aim of this study was to design and investigate the impacts of different concentrations of hesperidin, a glycoside flavonoid, on the in vitro preantral follicle growth and maturation in the three-dimensional (3D) culture system which was made with sodium alginate. Preantral follicles (n = 1363) were mechanically isolated from immature mice ovaries, then, after capsulating, they were randomly divided into four groups: the control group received no concentration of hesperidin, and three experimental groups were supplemented with 10, 22.5, and 50 µmol/L of hesperidin. All groups were cultured for 12 days. At the end of the culture period, the percentage of survival rate, antrum formation, obtained metaphase II oocytes, and the secretion of 17β-estradiol and progesterone were significantly higher in the group Hesp 50 (50 µmol/L hesperidin). Moreover, the mean average of follicular diameter cultured in the group Hesp 50 was also increased and the mRNA expression levels of PCNA, FSH-R, and Bcl-2 genes were higher, while Bax mRNA expression was significantly reduced compared with the other groups. Follicles cultured in the presence of 50 µmol/L of hesperidin had a higher fertilization rate and embryo development. Adding hesperidin at the concentration of 50 µmol/L to the culture medium resulted in higher follicular growth and maturation and increased the rate of in vitro fertilization and embryo development. Impact statement: It has been stated that hesperidin has many pharmacological effects, such as anti-inflammatory and antioxidant effects, antimicrobial activity, and anti-carcinogenic activity; but hesperidin and its derivatives have been under investigation as anti-fertility factors for a very long time. However, our results show that hesperidin can improve mice follicular growth and maturation during in vitro 3D culture. Hesperidin as an antioxidant factor could enhance the mRNA expression levels of two important genes involved in folliculogenesis, PCNA, and FSH-R. Our results prove for the first time that hesperidin not only has deleterious effects on follicular development but can also increase rates of in vitro fertilization and embryo development. © 2019 by the Society for Experimental Biology and Medicine

Reactive oxygenated species (ROS) in male fertility; source, interaction mechanism and antioxidant therapy

Currently ROS is known as the cause of many diseases and damage to cells and tissues. Many studies have shown that ROS is related to diseases such as cancer, cardiovascular, diabetic nephropathy and even processes such as aging and infertility. The studies of the past two decades reveal that low and controlled ROS values in the physiological process of the cell are secondary; while their physiological values is essential for the normal activity of the cell. Studies show that in most cells, the physiological amounts of ROS are produced by NADPH oxidase family enzymes. These enzymes are present in the plasma membrane of the cells, where they increase the production of ROS by connecting them with calcium. ROS in sperm also plays different physiological roles from the time of its production to the time of its emergence with an oocyte, but the pathological effects of its excessive production are also evident. This includes increasing the damage to DNA and increasing sperm apoptosis, which is responsible for including infertility in an important part of interfile men. In the infertility treatment clinics, that their number of clients is increasing every day, sperms find fertilization opportunity with the oocyte in the culture medium; but prior to that, sperm should passes different washing steps. These washing processes increase ROS production. The production of ROS is increased by the method of separating the sperm by centrifuging and then allowing sperm moves to float. This is one of the common methods in infertility treatment centers for sperm washing, while the sperm in washing stages has been deprived of its antioxidant source. Therefore, the use of vitamins either orally or in a sperm medium can increase the chances of fertilization and fertility of infertile individuals by preventing the motility of sperm and preventing their increased mortality and reducing DNA damage. © RJPT All right reserved

Reactive oxygenated species (ROS) in male fertility; source, interaction mechanism and antioxidant therapy

Currently ROS is known as the cause of many diseases and damage to cells and tissues. Many studies have shown that ROS is related to diseases such as cancer, cardiovascular, diabetic nephropathy and even processes such as aging and infertility. The studies of the past two decades reveal that low and controlled ROS values in the physiological process of the cell are secondary; while their physiological values is essential for the normal activity of the cell. Studies show that in most cells, the physiological amounts of ROS are produced by NADPH oxidase family enzymes. These enzymes are present in the plasma membrane of the cells, where they increase the production of ROS by connecting them with calcium. ROS in sperm also plays different physiological roles from the time of its production to the time of its emergence with an oocyte, but the pathological effects of its excessive production are also evident. This includes increasing the damage to DNA and increasing sperm apoptosis, which is responsible for including infertility in an important part of interfile men. In the infertility treatment clinics, that their number of clients is increasing every day, sperms find fertilization opportunity with the oocyte in the culture medium; but prior to that, sperm should passes different washing steps. These washing processes increase ROS production. The production of ROS is increased by the method of separating the sperm by centrifuging and then allowing sperm moves to float. This is one of the common methods in infertility treatment centers for sperm washing, while the sperm in washing stages has been deprived of its antioxidant source. Therefore, the use of vitamins either orally or in a sperm medium can increase the chances of fertilization and fertility of infertile individuals by preventing the motility of sperm and preventing their increased mortality and reducing DNA damage. © RJPT All right reserved

The global, regional, and national burden of stomach cancer in 195 countries, 1990�2017: a systematic analysis for the Global Burden of Disease study 2017

Background Stomach cancer is a major health problem in many countries. Understanding the current burden of stomach cancer and the differential trends across various locations is essential for formulating effective preventive strategies. We report on the incidence, mortality, and disability-adjusted life-years (DALYs) due to stomach cancer in 195 countries and territories from 21 regions between 1990 and 2017. Methods Estimates from GBD 2017 were used to analyse the incidence, mortality, and DALYs due to stomach cancer at the global, regional, and national levels. The rates were standardised to the GBD world population and reported per 100 000 population as age-standardised incidence rates, age-standardised death rates, and age-standardised DALY rates. All estimates were generated with 95% uncertainty intervals (UIs). Findings In 2017, more than 1.22 million (95% UI 1.19-1.25) incident cases of stomach cancer occurred worldwide, and nearly 865 000 people (848 000-885 000) died of stomach cancer, contributing to 19.1 million (18.7-19.6) DALYs. The highest age-standardised incidence rates in 2017 were seen in the high-income Asia Pacific (29.5, 28.2-31.0 per 100 000 population) and east Asia (28.6, 27.3-30.0 per 100 000 population) regions, with nearly half of the global incident cases occurring in China. Compared with 1990, in 2017 more than 356 000 more incident cases of stomach cancer were estimated, leading to nearly 96 000 more deaths. Despite the increase in absolute numbers, the worldwide age-standardised rates of stomach cancer (incidence, deaths, and DALYs) have declined since 1990. The drop in the disease burden was associated with improved Socio-demographic Index. Globally, 38.2% (21.1-57.8) of the age-standardised DALYs were attributable to high-sodium diet in both sexes combined, and 24.5% (20.0-28.9) of the age-standardised DALYs were attributable to smoking in males. Interpretation Our findings provide insight into the changing burden of stomach cancer, which is useful in planning local strategies and monitoring their progress. To this end, specific local strategies should be tailored to each country's risk factor profile. Beyond the current decline in age-standardised incidence and death rates, a decrease in the absolute number of cases and deaths will be possible if the burden in east Asia, where currently almost half of the incident cases and deaths occur, is further reduced

The global, regional, and national burden of stomach cancer in 195 countries, 1990�2017: a systematic analysis for the Global Burden of Disease study 2017

Background: Stomach cancer is a major health problem in many countries. Understanding the current burden of stomach cancer and the differential trends across various locations is essential for formulating effective preventive strategies. We report on the incidence, mortality, and disability-adjusted life-years (DALYs) due to stomach cancer in 195 countries and territories from 21 regions between 1990 and 2017. Methods: Estimates from GBD 2017 were used to analyse the incidence, mortality, and DALYs due to stomach cancer at the global, regional, and national levels. The rates were standardised to the GBD world population and reported per 100 000 population as age-standardised incidence rates, age-standardised death rates, and age-standardised DALY rates. All estimates were generated with 95 uncertainty intervals (UIs). Findings: In 2017, more than 1·22 million (95 UI 1·19�1·25) incident cases of stomach cancer occurred worldwide, and nearly 865 000 people (848 000�885 000) died of stomach cancer, contributing to 19·1 million (18·7�19·6) DALYs. The highest age-standardised incidence rates in 2017 were seen in the high-income Asia Pacific (29·5, 28·2�31·0 per 100 000 population) and east Asia (28·6, 27·3�30·0 per 100 000 population) regions, with nearly half of the global incident cases occurring in China. Compared with 1990, in 2017 more than 356 000 more incident cases of stomach cancer were estimated, leading to nearly 96 000 more deaths. Despite the increase in absolute numbers, the worldwide age-standardised rates of stomach cancer (incidence, deaths, and DALYs) have declined since 1990. The drop in the disease burden was associated with improved Socio-demographic Index. Globally, 38·2 (21·1�57·8) of the age-standardised DALYs were attributable to high-sodium diet in both sexes combined, and 24·5 (20·0�28·9) of the age-standardised DALYs were attributable to smoking in males. Interpretation: Our findings provide insight into the changing burden of stomach cancer, which is useful in planning local strategies and monitoring their progress. To this end, specific local strategies should be tailored to each country's risk factor profile. Beyond the current decline in age-standardised incidence and death rates, a decrease in the absolute number of cases and deaths will be possible if the burden in east Asia, where currently almost half of the incident cases and deaths occur, is further reduced. Funding: Bill & Melinda Gates Foundation. © 2020 The Author(s). Published by Elsevier Ltd. This is an Open Access article under the CC BY 4.0 licens

The global, regional, and national burden of oesophageal cancer and its attributable risk factors in 195 countries and territories, 1990–2017: a systematic analysis for the Global Burden of Disease Study 2017

Background Oesophageal cancer is a common and often fatal cancer that has two main histological subtypes: oesophageal squamous cell carcinoma and oesophageal adenocarcinoma. Updated statistics on the incidence and mortality of oesophageal cancer, and on the disability-adjusted life-years (DALYs) caused by the disease, can assist policy makers in allocating resources for prevention, treatment, and care of oesophageal cancer. We report the latest estimates of these statistics for 195 countries and territories between 1990 and 2017, by age, sex, and Socio-demographic Index (SDI), using data from the Global Burden of Diseases, Injuries, and Risk Factors Study 2017 (GBD). Methods We used data from vital registration systems, vital registration-samples, verbal autopsy records, and cancer registries, combined with relevant modelling, to estimate the mortality, incidence, and burden of oesophageal cancer from 1990 to 2017. Mortality-to-incidence ratios (MIRs) were estimated and fed into a Cause of Death Ensemble model (CODEm) including risk factors. MIRs were used for mortality and non-fatal modelling. Estimates of DALYs attributable to the main risk factors of oesophageal cancer available in GBD were also calculated. The proportion of oesophageal squamous cell carcinoma to all oesophageal cancers was extracted by use of publicly available data, and its variation was examined against SDI, the Healthcare Access and Quality (HAQ) Index, and available risk factors in GBD that are specific for oesophageal squamous cell carcinoma (eg, unimproved water source and indoor air pollution) and for oesophageal adenocarcinoma (gastro-oesophageal reflux disease). Findings There were 473 000 (95% uncertainty interval [95% UI] 459 000–485 000) new cases of oesophageal cancer and 436 000 (425 000–448 000) deaths due to oesophageal cancer in 2017. Age-standardised incidence was 5·9 (5·7–6·1) per 100 000 population and age-standardised mortality was 5·5 (5·3–5·6) per 100 000. Oesophageal cancer caused 9·78 million (9·53–10·03) DALYs, with an age-standardised rate of 120 (117–123) per 100 000 population. Between 1990 and 2017, age-standardised incidence decreased by 22·0% (18·6–25·2), mortality decreased by 29·0% (25·8–32·0), and DALYs decreased by 33·4% (30·4–36·1) globally. However, as a result of population growth and ageing, the total number of new cases increased by 52·3% (45·9–58·9), from 310 000 (300 000–322 000) to 473 000 (459 000–485 000); the number of deaths increased by 40·0% (34·1–46·3), from 311 000 (301 000–323 000) to 436 000 (425 000–448 000); and total DALYs increased by 27·4% (22·1–33·1), from 7·68 million (7·42–7·97) to 9·78 million (9·53–10·03). At the national level, China had the highest number of incident cases (235 000 [223 000–246 000]), deaths (213 000 [203 000–223 000]), and DALYs (4·46 million [4·25–4·69]) in 2017. The highest national-level age-standardised incidence rates in 2017 were observed in Malawi (23·0 [19·4–26·5] per 100 000 population) and Mongolia (18·5 [16·4–20·8] per 100 000). In 2017, age-standardised incidence was 2·7 times higher, mortality 2·9 times higher, and DALYs 3·0 times higher in males than in females. In 2017, a substantial proportion of oesophageal cancer DALYs were attributable to known risk factors: tobacco smoking (39·0% [35·5–42·2]), alcohol consumption (33·8% [27·3–39·9]), high BMI (19·5% [6·3–36·0]), a diet low in fruits (19·1% [4·2–34·6]), and use of chewing tobacco (7·5% [5·2–9·6]). Countries with a low SDI and HAQ Index and high levels of indoor air pollution had a higher proportion of oesophageal squamous cell carcinoma to all oesophageal cancer cases than did countries with a high SDI and HAQ Index and with low levels of indoor air pollution. Interpretation Despite reductions in age-standardised incidence and mortality rates, oesophageal cancer remains a major cause of cancer mortality and burden across the world. Oesophageal cancer is a highly fatal disease, requiring increased primary prevention efforts and, possibly, screening in some high-risk areas. Substantial variation exists in age-standardised incidence rates across regions and countries, for reasons that are unclear. Funding Bill & Melinda Gates Foundation