Detecting Somatic Mutations in Normal Cells

Somatic mutations have been studied extensively in the context of cancer. Recent studies have demonstrated that high-throughput sequencing data can be used to detect somatic mutations in non-tumor cells. Analysis of such mutations allows us to better understand the mutational processes in normal cells, explore cell lineages in development, and examine potential associations with age-related disease. We describe here approaches for characterizing somatic mutations in normal and non-tumor disease tissues. We discuss several experimental designs and common pitfalls in somatic mutation detection, as well as more recent developments such as phasing and linked-read technology. With the dramatically increasing numbers of samples undergoing genome sequencing, bioinformatic analysis will enable the characterization of somatic mutations and their impact on non-cancer tissues.

Pulmonary Capillary Hemorrhage Induced by Different Imaging Modes of Diagnostic Ultrasound

The induction of pulmonary capillary hemorrhage (PCH) is a well-established non-thermal biological effect of pulsed ultrasound in animal models. Typically, research has been done using laboratory pulsed ultrasound systems with a fixed beam and, recently, by B-mode diagnostic ultrasound. In this study, a GE Vivid 7 Dimension ultrasound machine with 10 L linear array probe was used at 6.6 MHz to explore the relative PCH efficacy of B-mode imaging, M-mode (fixed beam), color angio mode Doppler imaging and pulsed Doppler mode (fixed beam). Anesthetized rats were scanned in a warmed water bath, and thresholds were determined by scanning at different power steps, 2 dB apart, in different groups of six rats. Exposures were performed for 5 min, except for a 15-s M-mode group. Peak rarefactional pressure amplitude thresholds were 1.5 MPa for B-mode and 1.1 MPa for angio Doppler mode. For the non-scanned modes, thresholds were 1.1 MPa for M-mode and 0.6 MPa for pulsed Doppler mode with its relatively high duty cycle (7.7 × 10 -3 vs. 0.27 × 10 -3 for M-mode). Reducing the duration of M-mode to 15 s (from 300 s) did not significantly reduce PCH (area, volume or depth) for some power settings, but the threshold was increased to 1.4 MPa. Pulmonary sonographers should be aware of this unique adverse bio-effect of diagnostic ultrasound and should consider reduced on-screen mechanical index settings for potentially vulnerable patients.

Ultrasonic Cavitation-Enabled Treatment for Therapy of Hypertrophic Cardiomyopathy: Proof of Principle

Ultrasound myocardial cavitation-enabled treatment was applied to the SS-16 BN rat model of hypertrophic cardiomyopathy for proof of the principle underlying myocardial reduction therapy. A focused ultrasound transducer was targeted using 10-MHz imaging (10 S, GE Vivid 7) to the left ventricular wall of anesthetized rats in a warmed water bath. Pulse bursts of 4-MPa peak rarefactional pressure amplitude were intermittently triggered 1:8 heartbeats during a 10-min infusion of a microbubble suspension. Methylprednisolone was given to reduce initial inflammation, and Losartan was given to reduce fibrosis in the healing tissue. At 28 d post therapy, myocardial cavitation-enabled treatment significantly reduced the targeted wall thickness by 16.2% (p <0.01) relative to shams, with myocardial strain rate and endocardial displacement reduced by 34% and 29%, respectively, which are sufficient for therapeutic treatment. Premature electrocardiogram complexes and plasma troponin measurements were found to identify optimal and suboptimal treatment cohorts and would aid in achieving the desired impact. With clinical translation, myocardial cavitation-enabled treatment should fill the need for a new non-invasive hypertrophic cardiomyopathy therapy option.

Pulmonary Capillary Hemorrhage Induced by Diagnostic Ultrasound in Ventilated Rats

Pulmonary capillary hemorrhage (PCH) can be induced by diagnostic ultrasound-a potential safety issue. Anesthetized rats were intubated for intermittent positive-pressure ventilation (IPPV) with 0 end-expiratory pressure, +4 cm H 2 O end-expiratory pressure (PEEP) and -4 cm H 2 O end-expiratory pressure (NEEP). Rats were imaged at 7.6 MHz with a Philips HDI 5000 ultrasound machine. The output was low (mechanical index [MI] = 0.22) for aiming and then was raised for 5 min in 20 different exposure groups with n = 8. Peak rarefactional pressure amplitudes were measured in water and de-rated for chest attenuation. The PCH areas were measured on the lung surface. At 2.2 MPa, PCH was 9.3 ± 6.6 mm 2 for IPPV, 1.6 ± 3.2 mm 2 for PEEP (p <0.001) and 26.8 ± 6.4 mm 2 for NEEP (p <0.001). Thresholds were 1.3 MPa for IPPV, 2.1 MPa for PEEP and 1.0 MPa for NEEP. The small ventilator pressures subtracted or added to trans-capillary stress generated by diagnostic ultrasound pulses, virtually eliminating PCH for PEEP but enhancing PCH for NEEP.

Influence of Microbubble Size and Pulse Amplitude on Hepatocyte Injury Induced by Contrast-Enhanced Diagnostic Ultrasound

Recent research has found that contrast-enhanced diagnostic ultrasound (CEDUS) has the potential to induce localized injury in the liver, with clearly observable effects for contrast agent doses higher than the recommended dose and maximal mechanical index values. This study was undertaken to assess effects with intermittent exposure at lower contrast doses of infusion and at reduced output to determine thresholds. In addition, microbubble (MB) suspensions with enhanced content of larger MBs were tested. Exposure from a phased array probe (GE Vivid 7 Dimension, GE Vingmed Ultrasound, Horten, Norway) was applied at 1.6 MHz and 1-s intermittent frame trigger for 10 min with infusion of MB suspension with normal (1.8 µm), medium (3.1 µm) and large (5.3 µm) mean MB diameters. The bio-effect endpoint was the count of hepatocytes stained with Evans blue dye in frozen sections. For the normal MBs, the count increased for clinically relevant infusion dosages, but leveled off above 20 µL/kg/min. The evidence of injury declined with time from 30 min to 4 h and was lacking at 24 h. The exposure thresholds in terms of peak rarefactional pressure amplitude, divided by the square root of frequency (in situ mechanical index) were 1.7, 1.3 and 1.2 for the normal-, medium- and large-sized MB suspensions. The enhanced efficacy for larger MBs lends support to the two-criterion model for cavitational microvascular injury during CEDUS. Overall, CEDUS in liver appears to have markedly less potential for induction of tissue injury than has been reported in other tissues, which indicates a satisfactory safety profile for CEDUS using recommended parameters in normal liver.

Effect of optimal combination of Huangqi (Radix Astragali Mongolici) and Ezhu (Rhizoma Curcumae Phaeocaulis) on proliferation and apoptosis of A549 lung cancer cells

OBJECTIVE To investigate the effect of optimal combination (E) of Huangqi (Radix Astragali Mongolici) and Ezhu (Rhizoma Curcumae Phaeocaulis) on proliferation and apoptosis of A549 lung cancer cells and the possible mechanism underpinning the action. METHODS A uniform design method was used to optimize the E of Huangqi (Radix Astragali Mongolici) and Ezhu (Rhizoma Curcumae Phaeocaulis) in A549 lung cancer cells. MTS assay was applied to analyze the effect of the component formula of Huangqi (Radix Astragali Mongolici) and Ezhu (Rhizoma Curcumae Phaeocaulis) on A549 cells viability in various uniform design groups. A549 cells with exponential growth in routine culture were exposed to CoCl2 (200 μmol/L) to mimic hypoxic conditions. Group 0 was treated with RPMI-1640, the group CoCl2 was treated with CoCl2 (200 μmol/L), the group DDP + CoCl2 was treated with 4 mg/L Cisplatin injection (DDP) + CoCl2 (200 μmol/L), and the drug group was treated with various dose of E (0.5E, 1E, 2E) + CoCl2 (200 μmol/L). All groups were cultured for 24 h. Cell apoptosis was measured by Annexin V-FITC/propidium iodide double staining and flow cytometry. Western blot assay and quantitative real-time polymerase chain reaction (qRT-PCR) were employed to detect the protein and mRNA expression of B-celllymphoma-2 (Bcl-2), Bcl-2-associated X protein (Bax) and cysteinyl aspartate specific proteinase-3 (caspase-3). RESULTS The E obtained by the uniform design was comprise of 200 mg/L Astragalus polysaccharide (X1) and 32 mg/L Curcumin (X3). Group DDP+ CoCl2, group 1E + CoCl2 and group 2E + CoCl2 promoted the apoptosis of A549 cells (P 0.05). Compared with group 0, various doses of E + CoCl2 could up-regulate the expression of Bax and caspase-3 and down-regulate the expression of Bcl-2 at protein and mRNA levels (P CONCLUSION Astragalus polysaccharide and Curcumin was the optimal combination of Huangqi (Radix Astragali Mongolici) and Ezhu (Rhizoma Curcumae Phaeocaulis). E promoted the apoptosis of A549 cells. Combination of Astragalus polysaccharide and Curcumin increased the expression of Bax and caspase-3, and decreased the expression of Bcl-2 to initiate apoptosis in A549 cells under chemical-induced hypoxia.

Effects of long-term green manure application on the content and structure of dissolved organic matter in red paddy soil

Adiposity and risk of ischaemic and haemorrhagic stroke in 0·5 million Chinese men and women: a prospective cohort study

China has high stroke rates despite the population being relatively lean. Uncertainty persists about the relevance of adiposity to risk of stroke types. We aimed to assess the associations of adiposity with incidence of stroke types and effect mediation by blood pressure in Chinese men and women.

Age-specific association between blood pressure and vascular and non-vascular chronic diseases in 0·5 million adults in China: a prospective cohort study

The age-specific association between blood pressure and vascular disease has been studied mostly in high-income countries, and before the widespread use of brain imaging for diagnosis of the main stroke types (ischaemic stroke and intracerebral haemorrhage). We aimed to investigate this relationship among adults in China.

Smoking and smoking cessation in relation to risk of diabetes in Chinese men and women: a 9-year prospective study of 0·5 million people

In developed countries, smoking is associated with increased risk of diabetes. Little is known about the association in China, where cigarette consumption has increased (first in urban, then in rural areas) relatively recently. Moreover, uncertainty remains about the effect of smoking cessation on diabetes in China and elsewhere. We aimed to assess the associations of smoking and smoking cessation with risk of incident diabetes among Chinese adults.