Assessment of Cerebral White Matter Involvement in Amyotrophic Lateral Sclerosis Patients With Disease Progression and Cognitive Impairment by Fixel-Based Analysis and Neurite Orientation Dispersion and Density Imaging.

BACKGROUND: Previous studies using emerging diffusion MRI techniques have revealed damage to the white matter (WM) microstructure in amyotrophic lateral sclerosis (ALS), particularly the influence of crossed fibers, but there is a lack of subgroup analyses. PURPOSE: To detect WM microstructural changes in ALS patients using fixel-based analysis (FBA) and neurite orientation dispersion and density imaging (NODDI) MRI. STUDY TYPE: Prospective. POPULATION: Thirty-six ALS patients (aged 60.50 ± 9.5 years) and 25 healthy controls (HCs) (aged 58.90 ± 8.1 years). FIELD STRENGTH/SEQUENCE: 3 T; NODDI and FBA (b-values = 0, 1000, and 2500 seconds/mm2 ). ASSESSMENT: Subgroups were performed according to progression rate and cognition, including fast and slow progression (FP/SP), ALS with and without cognitive impairment (ALS-ci/ALS-nci). Fiber density (FD), fiber-bundle cross-section (FC), combined fiber density and cross-section (FDC), neurite density index (NDI), orientation dispersion index (ODI), isotropic volume fraction (ISO), and fractional anisotropy (FA) were calculated and their correlation with clinical variables examined. STATISTICAL TESTING: Chi-square test, Mann-Whitney U test, two-sample t test, partial correlation analysis, and false discovery rate (FDR) corrected. A P-value <0.05 was considered significant. RESULTS: ALS patients had lower FD and FDC values predominantly in the corticospinal tract (CST) and corpus callosum (CC) regions, as well as lower NDI value in the CC, radial crown, and internal capsule compared to HCs. Subgroup analysis based on progression rate and cognitive function showed significant differences in FBA results. The FC in the right CST region was significantly lower in the FP than SP, and the FD in the CC region was significantly lower in the ALS-ci than ALS-nci. Furthermore, a negative correlation was found between the mean FC value and the rate of progression in ALS patients (r = -0.408). DATA CONCLUSION: FBA is a powerful tool for detecting complex cerebral WM microstructural damage for evaluating ALS cognition and disease progression.

A cost efficient protocol to introduce epitope tags by CRISPR-Cas9 mediated gene knock-in with asymmetric semi-double stranded template.

To study the biological function of uncharacterized proteins, specific antibodies are in high demand. However, the production of desirable antibodies such as highly specific or high affinity is not always successful. Furthermore, even if commercially available antibodies exist, the cost, quality, and accessibility often differ from country to country. In comparison, epitope tags are reliable and economical options since good antibodies against major epitope tags are commercially available. Although exogenously expressed epitope-tagged protein appears as a timely method, the excessive protein production may not faithfully recapitulate its biology. Thanks to the recent advances in genome editing by CRISPR-Cas9, HDR-mediated endogenous protein tagging has become an accessible approach for many labs. However, currently the synthesis of long (>100 bp), chemically modified oligos can be time-consuming and costly. To develop a reliable, simple, and cost-effective epitope-tagging method that requires minimal materials and apparatus, we focus on an approach utilizing two non-chemically modified shorter-annealed oligos (semi-dsODNs) mediated HDR for epitope tags insertion. We also use a cationic lipid chemical, polyethyleneimine (PEI), for plasmid delivery to minimize the cost and materials used while a considerable success rate could be achieved. • This protocol provides a more economical way to generate CRISPR-Cas9 mediated gene knock-in. • This protocol provides a simplified design of semi-dsODN without chemical modification on the oligos. • This protocol provides a simplified experimental procedure. In vitro assembled Cas9 complex and electroporation are not required.

Optical palpation for tumor margin assessment in breast-conserving surgery.

Intraoperative margin assessment is needed to reduce the re-excision rate of breast-conserving surgery. One possibility is optical palpation, a tactile imaging technique that maps stress (force applied across the tissue surface) as an indicator of tissue stiffness. Images (optical palpograms) are generated by compressing a transparent silicone layer on the tissue and measuring the layer deformation using optical coherence tomography (OCT). This paper reports, for the first time, the diagnostic accuracy of optical palpation in identifying tumor within 1 mm of the excised specimen boundary using an automated classifier. Optical palpograms from 154 regions of interest (ROIs) from 71 excised tumor specimens were obtained. An automated classifier was constructed to predict the ROI margin status by first choosing a circle diameter, then searching for a location within the ROI where the circle was ≥ 75% filled with high stress (indicating a positive margin). A range of circle diameters and stress thresholds, as well as the impact of filtering out non-dense tissue regions, were tested. Sensitivity and specificity were calculated by comparing the automated classifier results with the true margin status, determined from co-registered histology. 83.3% sensitivity and 86.2% specificity were achieved, compared to 69.0% sensitivity and 79.0% specificity obtained with OCT alone on the same dataset using human readers. Representative optical palpograms show that positive margins containing a range of cancer types tend to exhibit higher stress compared to negative margins. These results demonstrate the potential of optical palpation for margin assessment.

[Profile Characteristics of VOCs from Wood and Economic Crop Burning].

Wood and economic crops are still widely used in rural areas of China. Although their combustion is an important source of volatile organic compounds (VOCs), study on their emission characteristics is relatively weak. In this study, three kinds of wood (poplar, cedarwood, and citrus branches) and six economic crop straws (soybean stalk, sesame stalk, corn cob, cotton stalk, peanut stalk, and corn stalk) were selected and their burning was simulated in the laboratory. A dilution tunnel system was used to dilute the smoke, and then Tedlar bags were used to collect the smoke. The compositions of 102 VOCs were analyzed by Agilent 7820A/5977E gas chromatography/mass spectrometry. The ozone formation potential (OFP) of VOCs for different types of biomass burning was analyzed. The results indicated that there are differences in the VOC compositions of different types of biomass burning emissions. Ethane (11.1%), trans-2-pentene (15.4%), ethylene (8.3%), and dichloromethane (11.9%) are the main VOCs emitted from poplar and cedarwood burning. Toluene (49.8%) is the most abundant species of VOC emitted from burning of citrus branches. Ethylene (11.8%-17.5%) and acetone (9.2%-14.7%) are the main VOCs components of straw burning. Corn stalks, peanut stalks, and citrus branches have similar VOC source profiles, with the coefficient of divergence less than 0.1. The benzene/toluene ratio for biomass burning emissions obtained in this study and in the literature is in the range of 0.030-6.48. It is arguable that a value higher than 1 indicated the impact of biomass burning. The contributions of alkenens, oxygenated VOCs, and aromatic hydrocarbons to the OFP of biomass burning were 30.6%-80.3%, 6.5%-21.0%, and 3.8%-56.5%, respectively. The components contributing more than 10.0% to the OFP are ethylene, propylene, trans-2-pentene, cis-2-pentene, toluene, and propionaldehyde.

Diagnostic Accuracy of Quantitative Micro-Elastography for Margin Assessment in Breast-Conserving Surgery.

Inadequate margins in breast-conserving surgery (BCS) are associated with an increased likelihood of local recurrence of breast cancer. Currently, approximately 20% of BCS patients require repeat surgery due to inadequate margins at the initial operation. Implementation of an accurate, intraoperative margin assessment tool may reduce this re-excision rate. This study determined, for the first time, the diagnostic accuracy of quantitative micro-elastography (QME), an optical coherence tomography (OCT)-based elastography technique that produces images of tissue microscale elasticity, for detecting tumor within 1 mm of the margins of BCS specimens. Simultaneous OCT and QME were performed on the margins of intact, freshly excised specimens from 83 patients undergoing BCS and on dissected specimens from 7 patients undergoing mastectomy. The resulting three-dimensional images (45 × 45 × 1 mm) were coregistered with postoperative histology to determine tissue types present in each scan. Data from 12 BCS patients and the 7 mastectomy patients served to build a set of images for reader training. One hundred and fifty-four subimages (10 × 10 × 1 mm) from the remaining 71 BCS patients were included in a blinded reader study, which resulted in 69.0% sensitivity and 79.0% specificity using OCT images, versus 92.9% sensitivity and 96.4% specificity using elasticity images. The quantitative nature of QME also facilitated development of an automated reader, which resulted in 100.0% sensitivity and 97.7% specificity. These results demonstrate high accuracy of QME for detecting tumor within 1 mm of the margin and the potential for this technique to improve outcomes in BCS. SIGNIFICANCE: An optical imaging technology probes breast tissue elasticity to provide accurate assessment of tumor margin involvement in breast-conserving surgery.

H-Type Hypertension Is a Risk Factor for Cerebral Small-Vessel Disease.

BACKGROUND: The correlation between H-type hypertension and cerebral small-vessel diseases (CSVD) remains uncertain. OBJECTIVE: The present study was designed to explore the possible relationship between H-type hypertension and CSVD spectrum and total burden. METHOD: We included 329 patients in the present study and divided them into four groups: the H-type hypertension group, isolated hypertension group, isolated hyperhomocysteinemia (HHcy) group, and control group. Clinical variables of interest and the MR examination sequences were obtained. We counted the presence of each CSVD feature and rated the total burden of CSVD on an ordinal scale from 0 to 4 according to a recent described score rule. RESULT: The results showed that H-type hypertension was associated with the presence of cerebral microbleeds (CMBs), and the severity of white-matter hyperintensities (WMHs) and peripheral vascular space (PVS). CSVD total burden was significantly related to age (OR: 1.059, 95% CI: 1.037-1.082), systolic pressure (OR: 1.122, 95% CI: 1.007-1.136), triglycerides (OR: 1.386, 95% CI: 1.037-1.854), isolated HHcy (OR: 4.154, 95% CI 1.836-9.401), and H-type hypertension (OR: 5.028, 95% CI: 2.323-10.883). Also, we further observed hypertension and HHcy had a synergistic effect on CSVD total burden (OR: 2.776, 95% CI: 1.564-4.927). CONCLUSION: H-type hypertension was associated with CSVD total burden and CSVD spectrum, which deserves further prevention measures. Furthermore, hypertension and HHcy had a synergistic effect on CSVD total burden.

Clinical Utility of the Inflammatory Factors Combined With Lipid Markers in the Diagnostic and Prognostic Assessment of Ischemic Stroke: Based on Logistic Regression Models.

BACKGROUND: In this study, we developed novel logistic regression models for the diagnostic and prognostic assessment of ischemic stroke. METHODS: A total of 288 ischemic stroke patients and 300 controls admitted to The First Affiliated Hospital of Soochow University were included in the testing group. Two validation groups from The Affiliated Kunshan Hospital of Jiangsu University and The Second Affiliated Hospital of Soochow University were included to assess our novel assessment models. RESULTS: Results from the testing group indicated that the diagnostic assessment model for ischemic stroke prediction was: Logit(P) = 437.116 - 87.329 (Hypertension) - 89.700 (Smoking history) - 87.427 (Family history of ischemic stroke) - .090 (high-density lipoprotein cholesterol [HDL-C]) - 1.984 (low-density lipoprotein cholesterol [LDL-C]) - 17.005 (Lp(a)) - 15.486 (Apo A/Apo B), and the final prognostic assessment model of ischemic stroke was: Logit(P) = 458.437-92.343 (Hypertension) - 89.763 (Smoking history) + .251 (NLR) - .088 (HDL-C) - 1.994 (LDL-C) - 2.883 (hs-CRP) - .058 (IL-6) - 6.356 (TNF-α) - 16.485 (Lp(a)) - 17.658 (Apo A/Apo B). In the validation groups, our novel diagnostic assessment model showed good identification (with 87.5% sensitivity and 84.2% specificity in The Affiliated Kunshan Hospital of Jiangsu University, with 85.5% sensitivity and 89.0% specificity in The Second Affiliated Hospital of Soochow University). Moreover, our novel prognostic assessment model has a high value in identifying poor prognosis patients in the validation groups from The Affiliated Kunshan Hospital of Jiangsu University (χ2 = 8.461, P = .004), and The Second Affiliated Hospital of Soochow University (χ2 = 7.844, P = .005). CONCLUSIONS: The diagnostic and prognostic assessment models we have established are of great value in the diagnosis and prognostic evaluation of ischemic stroke.

Safety and cost-effectiveness thrombolysis by diffusion-weighted imaging and fluid attenuated inversion recovery mismatch for wake-up stroke.

Wake-up stroke, defined as patients who wake up with stroke symptoms which were not present prior to falling asleep, accounted for 14%-25% of acute ischemic stroke. Due to the unknown time of symptom onset, wake-up stoke was not in including criteria of intravenous thrombolysis. Several large randomized stroke trials using diffusion-weighted imaging(DWI)and fluid attenuated inversion recovery(FLAIR)mismatch patient selection may identify a subset of patients with wake-up stroke that can safely and effectively benefit from intravenous thrombolysis. In addition, economic factor was another important limitation to generalize thrombolysis treatment. Fortunately, MRI-based thrombolysis was a cost-effective treatment for wake-up stroke compared to these patients with no thrombolysis.

A comparison between the pectoralis major myocutaneous flap and the free anterolateral thigh perforator flap for reconstruction in head and neck cancer patients: assessment of the quality of life.

Our study investigated the quality of life (QoL) of Chinese patients after immediate reconstruction surgery on individuals with head and neck cancer. In addition, we compared the differences between pectoralis major myocutaneous flap (PMMF) and anterolateral thigh free flap (ALTFF). The University of Washington Quality of Life questionnaire, version 4, was used to assess the QoL. Assessments were performed at least 24 months postoperatively. A total of 110 patients' records were obtained. Among them, 86 patients completed a QoL questionnaire (78.2%). No significant differences could be found in age, primary site, T stage, N stage, and postoperative radiotherapy between PMMF and ALTFF groups. However, there were significant differences between both groups in sex, operation time, and complication. A matched analysis was performed to compare the differences in QoL between patients with head and neck cancers reconstructed with PMMF or ALTFF. Patients reconstructed with ALTFF had better shoulder but worse speech functions. There was a significant effect on the QoL of head and neck cancer patients who had undergone either PMMF or ALTFF reconstruction. The result of this study provide useful information for physicians and patients during their discussion of treatment modalities for head and neck cancers.

Free fibula flap: assessment of quality of life of patients with head and neck cancer who have had defects reconstructed.

This study investigated the quality of life (QoL) of patients with head and neck cancer undergoing immediate reconstruction of the mandible with free fibula flap. From March 2006 to January 2011, the QoL of 42 patients was assessed using the Medical Outcomes Study Short Form 36 and the University of Washington QoL (version 4) questionnaires. The assessments were performed at least 24 months after surgery. A total of 31 of the 42 questionnaires (73.8%) were returned. The length of harvested fibula varied from 17.5 to 26.1 cm. In the Short Form 36, the lowest-scoring domain was vitality, whereas the highest scores occurred in physical role. According to the University of Washington QoL, the key domains affected by surgery are chewing, speech, and appearance. The domain of pain has the best score. There was a significant effect on the QoL of patients with head and neck cancer with resections of the mandible who had undergone free fibula flap reconstruction. Data from this study may provide useful information for physicians and patients, which may be of value during discussion of treatment modalities for head and neck cancers.

Assessment of the quality of life of patients with oral cancer after pectoralis major myocutaneous flap reconstruction with a focus on speech.

PURPOSE: The aim of the present study was to evaluate the quality of life (QoL) of patients with oral cancer who had undergone resection of the tongue and floor of the mouth and reconstruction with the pectoralis major flap. MATERIALS AND METHODS: The present study assessed 21 patients who had undergone pectoralis major flap reconstruction using the University of Washington QoL, version 4, questionnaire. A nonparametric Mann-Whitney U test was used to analyze the data. RESULTS: Of the 12 disease-specific domains, the best 3 scores from the patients were for pain, saliva, and anxiety, and the worst 3 scores were for taste, chewing, and swallowing. The mean UW-QoL composite score was 73.4. Swallowing was considered to be the most important issue within the previous 7 days, followed by chewing and speech. Those patients who had undergone wider excision had poorer speech. CONCLUSIONS: A significant effect was found on the QoL of patients with oral cancer who had undergone resection of the tongue and floor of the mouth with pectoralis major flap reconstruction.

[Cost-benefit analysis of comprehensive parasitic disease control project in Rongxian County, Guangxi Zhuang autonomous region].

OBJECTIVE: To evaluate the cost-benefit of comprehensive parasitic disease control project in Rongxian County of Guangxi, so as to provide the evidence for formulating the control strategy of soil-transmitted nematode. METHODS: The data of the overall input and output for soil-transmitted nematode control were collected from 2007 to 2009, and the cost-benefit was analyzed. RESULTS: The infection rates of soil-transmitted nematode dropped from 30.10% in 2007 to 12.30% in 2009. The overall cost of soil -transmitted nematode control was 969 185.6 Yuan from 2007 to 2009. The direct benefits arising from avoiding the incidence of the disease for populations reached 88.415 million Yuan in three years. The ratio of benefit-cost was 91:1. CONCLUSIONS: The soil -transmitted nematode infection brings a heavy financial burden on both the patients and society. The implementation of comprehensive control measures can effectively control the prevalence of the diseases and produce good social and economic benefits.

Effect of polymorphism of uncoupling protein 3 gene -55 (C>T) on the resting energy expenditure, total body fat and regional body fat in Chinese.

OBJECTIVE: To investigate the relationship of the C to T variant at the -55 site of the promoter region of uncoupling protein 3 gene (UCP3) with the resting energy expenditure and the parameters of body fat in Chinese population. METHODS: Three hundred Chinese (91 normal weight subjects, 209 overweight/obesity subjects) were genotyped for the UCP3 gene -55(C>T) by using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). Resting energy expenditure (REE), fat mass (FM), fat free mass (FFM) and the parameters for regional adipose tissue distribution were measured. RESULTS: Genotype frequencies of UCP3 gene -55(C>T) were not associated with obesity and different types of obesity. The REE level in normal weight subjects with TT homozygotes was higher than that in those with CT heterozygotes and CC homozygotes (P=0.0200). Similar tendency was also observed in overweight/obesity subjects. The FM/FFM exhibited significant difference between the overweight/obesity subjects with a TT genotype and those with a CT or CC genotype (P=0.0096). CONCLUSION: The level of difference in REE caused by the polymorphism of promoter region of UCP3 -55(C>T) may play a role in energy metabolism in Chinese.