[Correlations of diurnal brain functional variations with serum biomarkers and objective sleep quality in patients with chronic insomnia disorder].

Objectives: To investigate the correlations of diurnal brain functional variations with serum biomarkers and objective sleep quality in patients with chronic insomnia disorder (CID). Methods: A total of 60 CID patients and 30 healthy sleep volunteers who visited Department of Sleep Disorders of Chaohu Hospital affiliated to Anhui Medical University from March 2018 to June 2019 were collected. Diurnal brain function state was evaluated by Quantitative Measurement System of Brain Functional Status, and serum concentrations of corticotropin-releasing hormone (CRH) and glial fibrous acidic protein (GFAP) were detected using enzyme-linked immunosorbent assay (ELISA). The sleep quality in the CID group was evaluated using ploysomnography (PSG) at the same time. The brain function status indicators with significant changes in CID group were acquired, and the consistency of these indicators with serum biological markers and objective sleep quality was analyzed. Results: There were 23 males and 37 females in chronic in somnia patients; 15 males and 15 females in the healthy control group. Compared with the healthy controls, four brain function indicators of CID patients increased (brain inertia (196.0(163.0, 258.0)vs 168.5(148.8,182.5)), brain chaos (5.0(1.0, 10.0)vs 0(0,2.0)), internal concentration (31.0(13.0, 45.0)vs 2.0(0,27.5)) and endogenous anxiety (12.0(4.0, 18.0)vs 0(0,6.5)), but one indicator decreased (brain inhibitory value (47.0(32.0, 58.0)vs 59.0(46.3,66.3))) (all P<0.05).The brain chaos value positively correlated with the serum GFAP level (r=0.374,P=0.006), and the brain inertia value positively correlated with the serum CRH level (r=0.299,P=0.031). The value of brain inhibition positively correlated with the sleep latency (r=0.284,P=0.042). However, the values of internal concentration negatively correlated with the sleep efficiency (r=-0.276,P=0.048) and the time in non-rapid eye movement sleep stage 1 (r=-0.341, P=0.024). Conclusion: The brain waves of CID patients show significant changes in their brain function indicators, which are related to serum biological markers and objective sleep quality.

[Analysis on clustered regularly interspaced short palindromic repeats loci polymorphism of Yersinia pestis and its area distribution in Gansu province].

Objective: To study the clustered regularly interspaced short palindromic repeats (CRISPR) loci polymorphism of Yersinia pestis and its area distribution in Gansu province. Methods: A total of 203 strains of Yersinia pestis isolated from 1962 to 2014 were selected for the culture and extraction of DNA. Three pairs of CRISPR primers were used to amplify the strain DNA by PCR, and the PCR products were sequenced. The groups and genotypes of strains were determined according to the spacer and spacer arrangement of CRISPR loci in the strain. Cluster analysis was done by using the software BioNumerics 5.10. Results: A total of 16 spacers, including 9 species of YPa loci, 4 species of YPb loci and 3 species of YPc loci, were found in the 203 strains of Yersinia pestis. A new spacer of a1' was found. The 203 strains were divided into 5 CRISPR genotypes and classified into 5 CRISPR clusters (Cb2, Ca7, Ca7', CaΔ5' and Ca35'). Each cluster showed significant area-specific characteristics, Cb2 was mainly distributed in Huining country and Pingchuan district, Ca7 was mainly found in Aksai Kazak autonomous country, Ca7' was mainly found in Xiahe country, Ca35' was mainly found in Subei Mongolia autonomous county and Yumen city and CaΔ5' was mainly distributed in Sunan Yugur autonomous county. Conclusions: The strains from different plague foci in Gansu were distinguished by CRISPR, all kinds of clusters showed the obvious area specific characteristics. It is important to study the evolution of Yersinia pestis in Gansu and trace the molecular biology origin of human plague.

[Cardiopulmonary coupling analysis in diagnosis of sleep disorders].

Objectives: To compare diagnostic consistency for chronic insomnia (CI) and obstructive sleep apnea (OSA) between cardiopulmonary coupling (CPC) and polysomnographm (PSG). Methods: Two hundred and twenty-one patients were enrolled from the Department of Sleep Disorders, Chaohu Hospital affiliated to Anhui Medical University from July 2018 to December 2019, and monitored with overnight CPC and PSG simultaneously. According to clinical representations and PSG results, there were 88 males and 80 females with CI and OSA, including chronic insomnia (CI group, 93 cases), OSA (OSA group, 36 cases) and comorbid OSA with CI (COI group, 39 cases). The consistency of sleep and OSA parameters measured with CPC and PSG were analyzed. Results: (1)For all patients and CI group, the total sleep time (TST), sleep efficiency (SE) and rapid eye movement (REM) sleep time measured by CPC were significantly higher than those measured with PSG, and the wake time after sleep onset (WASO) was significantly lower than that measured with PSG (the specific median comparisons were as follows 420.0 min vs 395.5 min, 93.7% vs 81.8%, 90.0 min vs 37.5 min, 18.0 min vs 63.0 min in CI group, respectively; 414.0 min vs 392.5 min, 91.9% vs 81.9%, 72.0 min vs 34.8 min, 24.0 min vs 58.4 min in all patients, respectively (all P≤0.001). However, in the OSA patients, the TST, SE, WASO, REM sleep time and NREM sleep time measured using two methods were similar (all P>0.05). (2) According to OSA criteria, the consistency between CPC and PSG was fair (κ=0.255). Only CPC has a certain degree of value for OSA screening when the AHI ≥ 20/h (κ=0.580, sensitivity: 0.85, specificity: 0.82, positive predictive value: 0.59, negative predictive value: 0.95, positive likelihood ratio: 4.72). Conclusion: CPC technology may overestimate the sleep quality of CI patients, and its consistency is fair compared with that of PSG in the diagnosis of OSA.

[Effects of loupes and microscope on laminate veneer preparation].

OBJECTIVE: To assess and compare the effects of loupes and microscope on laminate veneer preparation of the first practitioner from the aspects of efficiency, quality and accuracy of preparation, and preference. METHODS: Twenty young prosthodontists from the Department of Prosthodontics, Peking University School and Hospital of Stomatology were recruited into this study, which was prospective, single blind, self-control trials. The participants had no experience of using dental magnification devices. They prepared laminate veneers in the artificial dental model, under routine visual field (control group), 2.5× headwear loupes (loupes group), and 8× operating microscope (microscopic group) by turning. The time for tooth preparation was recorded. Thereafter, subjective assessments of efficiency, quality of preparation and preference were performed by themselves using visual analogue score (VAS). Expert assessments of quality and accuracy of preparation were performed by two professors using stereomicroscope and digital technique respectively. RESULTS: In terms of efficiency, the subjective scores for the control group, loupes group and microscopic group were 7.15±1.73, 8.10±0.91 and 5.40±2.04, respectively. There was significant difference between the loupes group and microscopic group (P<0.05). The time of tooth preparation for the control group, loupes group and microscopic group was (430.10±163.04) s, (393.90±157.27) s and (441.95±164.18) s, respectively. There was significant difference between the loupes group and microscopic group (P<0.05). The loupes group was more efficient than the microscopic group. In terms of the quality of preparations, the subjective scores for the control group, loupes group and microscopic group were 6.55±2.09, 7.85±0.99 and 6.25±1.77, respectively. There was significant difference between the loupes group and microscopic group (P<0.05). The expert evaluations for the control group, loupes group and microscopic group were 12.20±1.67, 12.50±1.70 and 11.35±2.60, respectively. There was significant difference between the loupes group and microscopic group (P<0.05). The loupes group had higher quality than the microscopic group. In terms of the accuracy of preparations, the control group, loupes group and microscopic group of incisal 1/3 were (0.107±0.097) mm, (0.142±0.118) mm and (0.123±0.087) mm, respectively, of middle 1/3 were (0.128±0.073) mm, (0.113±0.105) mm and (0.125±0.077) mm, respectively, and of cervical 1/3 were (0.075±0.054) mm, (0.068±0.044) mm and (0.058±0.047) mm, respectively. There was no significant difference among the three groups (P>0.05). In terms of the preference, the subjective scores for the control group, loupes group and microscopic group were 6.55±2.31, 8.60±1.10 and 5.80±2.07, respectively. There was significant difference between the loupes group and microscopic group (P<0.05). The participants had the highest preference for loupes. CONCLUSION: For the first practitioners, loupes is better than microscope for laminate veneer preparation.

[Influence of implants prepared by selective laser melting on early bone healing].

OBJECTIVE: To evaluate the influence of the rough surface of dental implants prepared by selective laser melting (SLM) on early bone healing around titanium implants. METHODS: A total of sixteen titanium implants were involved in our research, of which eight implants were prepared by SLM (TIXOS Cylindrical, Leader-Novaxa, Milan, Italy; 3.3 mm×10 mm, internal hex) and the other eight were sandblasted, large-grit and acid-etched (SLA) implants (IMPLUS Cylindrical, Leader-Novaxa, Milan, Italy; 3.3 mm×10 mm, internal hex). All of the dental implants were inserted into the healed extraction sockets of the mandible of two adult male Beagle dogs. Half of the dental implants were designed to be healed beneath the mucosa and the other half were intended to be healed transgingivally and were immediately loaded by acrylic resin bridge restoration. Three types of tetracycline fluorescent labels, namely calcein blue, alizarin complexone and calcein, were administered into the veins of the Beagle dogs 2, 4, and 8 weeks after implant placement respectively for fluorescent evaluation of newly formed bone peri-implant. Both Beagle dogs were euthanized 12 weeks after implant insertion and the mandible block specimens containing the titanium implants and surrounding bone and soft tissue of each dog were carefully sectioned and dissected. A total of 16 hard tissue slices were obtained and stained with toluidine blue for microscopic examination and histomorphometric measurements. Histological observation was made for each slice under light microscope and laser scanning confocal microscope (LSCM). Comparison on new bone formation around titanium implants of each group was made and mineral apposition rate (MAR) was calculated for each group. RESULTS: Dental implants prepared by selective laser melting had achieved satisfying osseointegration to surrounding bone tissue after the healing period of 12 weeks. Newly formed bone tissue was observed creeping on the highly porous surface of the SLM implant and growing into the pores of surface structure. Higher MAR values were shown for SLM implants compared with SLA implants (P<0.01). CONCLUSION: Dental implants prepared by selective laser melting could promote early bone healing and improve mineral apposition rate.

[A novel tissue-engineered bone constructed by using human adipose-derived stem cells and biomimetic calcium phosphate scaffold coprecipitated with bone morphogenetic protein-2].

OBJECTIVE: To construct a novel biomimetic calcium phosphate (BioCaP) scaffold loaded with bone morphogenetic protein-2 (BMP-2), and to investigate its role in the osteogenesis of human adipose-derived stem cells (hASCs) in vitro and in vivo. METHODS: The BioCaP scaffold coprecipitated with BMP-2 (BMP-2-BioCaP) was constructed in this study. Field emission scanning electron microscopy (SEM) was used to analyze the morphology of the surfaces. The release kinetics was measured to evaluate the slow-release characteristics in vitro. BMP-2-BioCaP was immersed in proliferation medium (PM) or osteogenic medium (OM), respectively. The supernatants were collected and used to culture hASCs in vitro. Cell numbers were determined using the cell-counting kit-8 (CCK-8) to assess the cell proliferation. After 7 and 14 days, alkaline phosphatase (ALP) staining and quantification were performed to test the activity of ALP. After 14 and 21 days, the calcification deposition was determined by alizarin red S (ARS) staining and quantification. The expressions of the osteoblast-related genes were tested on day 4 and day 14. In the in vivo study, 6 nude mice were used and implanted subcutaneously into the back of the nude mice for 4 groups: (1) BioCaP scaffold only, (2) BioCaP scaffold+hASCs, (3) BMP-2-BioCaP scaffold, (4) BMP-2-BioCaP scaffold+hASCs (test group). After 4 weeks of implantation, hematoxylin-eosin (HE) staining was performed to evaluate the in vivo osteogenesis of hASCs. RESULTS: SEM observations showed that BioCaP and BMP-2-BioCaP scaffold were entirely composed of straight, plate-like and sharp-edged crystal units, and the length of the crystal units varied between 5 and 10 µm. Release kinetics analysis demonstrated that BMP-2 incorporated with BioCaP could be released at certain concentration and last for more than 21 days, and the accumulative protein release could reach 20%. CCK-8 assays showed that cell proliferation was not significantly affected by BMP-2-BioCaP. ALP activity was higher by the induction of OM+BMP-2-BioCaP than of the other groups (P<0.01). More mineralization deposition and more expressions of osteoblast-related genes such as Runt-related transcription factor 2 (RUNX2), ALP, osteopontin (OPN) and osteocalcin (OC) were determined in the OM+BMP-2-BioCaP group at different time points (P<0.01). HE staining showed that, in the test group and BMP-2-BioCaP scaffold group, the extracellular matrix (ECM) with eosinophilic staining were observed around hASCs, and newly-formed bone-like tissues could be found in ECM around the scaffold materials. Moreover, compared with the BMP-2-BioCaP scaffold group, more bone-like tissues could be observed in ECM with typical structure of bone tissue in the test groups. No obvious positive results were found in the other groups. CONCLUSION: BMP-2-BioCaP scaffold could achieve slow-release of BMP-2 and promote the osteogenic differentiation of hASCs in vitro and in vivo. The novel tissue-engineered bone composed of hASCs and BMP-2-BioCaPis promising for the repair of bone defect.

[The clinical management of co-occurring insomnia and obstructive sleep apnea hypopnea syndrome].

Insomnia and OSAHS are the comorbid diseases among the most common sleep disorders with high incidence. They both interact with each other apart from combined symptoms, therefore are defined as co-occurring insomnia and obstructive sleep apnea(CIO). CIO causes more serious damage to patients than single insomnia or OSAHS does, resulting in complex clinical treatments. However, the traditional treatment of CIO aims at solely OSAHS or insomnia patients with poor efficacy and compliance. Therefore, further investigation is required to elucidate the clinical mechanism and to optimize concurrent treatment for OSAHS and insomnia.

Effect and Mechanism of Portal Blood Stasis Removal on Intestinal Endotoxemia and Hepatic Ischemia Reperfusion Injury.

OBJECTIVE: We used a rabbit model of hepatic ischemia reperfusion in situ to observe the change of portal venous endotoxin level before reperfusion, and the effect of portal blood stasis removal on intestinal endotoxemia and hepatic ischemia reperfusion injury. The purpose was to find an ideal method for portal blood stasis removal and provide the experimental proof for clinical application of hepatectomy. METHODS AND MATERIALS: To investigate the effect of portal blood stasis removal on intestinal endotoxemia and hepatic ischemia reperfusion injury, a rabbit hepatic ischemia reperfusion injury model was established and treated with removal of portal blood stasis before the portal blood circulation was resumed. Serum endotoxin content, alanine aminotransferase (ALT), hyaluronic acid (HA), and content of malondialdehyde (MDA), tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), superoxide dismutase (SOD) and activation of nuclear factor-κB (NF-κB) in liver tissue were examined respectively. RESULTS: In portal blood stasis the level of serum endotoxin significantly decreased with each 2.5 mL blood removal (P < .01), subsequently reaching a minima at the 7.5 mL blood removal (P > .05). Removing portal blood stasis ameliorated endotoxemia and hepatic ischemia reperfusion injury as shown by ALT, HA, MDA, SOD, TNF-α, IL-6, and activation of NF-κB compared to no removal. The first 5 mL portal blood stasis contains high volume of endotoxin which may be responsible for hepatic reperfusion injury. CONCLUSION: Removal of portal blood stasis before the resume of splanchnic circulation may ameliorate intestinal endotoxemia and hepatic ischemia reperfusion injury.

Fengshui forests conserve genetic diversity: a case study of Phoebe bournei (Hemsl.) Yang in southern China.

Fengshui forests (sacred groves) are important in traditional Chinese culture and home to many endangered species. These forests may provide protection for some endangered plant species outside the nature reserves, but little is known about their role in genetic conservation. Using inter-simple sequence repeat (ISSR) markers, we compared the genetic diversity of 6 populations of Phoebe bournei (Hemsl.) Yang, a commercially important woody species, which is under second-class national protection and endemic to China. Samples were collected from the nature reserves and Fengshui forests in southern China. Herein, we show that Fengshui forest populations are capable of maintaining some level of genetic diversity. For nature reserve populations, the average NA and NE were 1.58 and 1.39, respectively; and for Fengshui forests, they were 1.39 and 1.12, respectively. For nature reserve populations, Nei's gene diversity (H) and Shannon's index (I) were 0.32 and 0.11, respectively; and for Fengshui forests, they were 0.22 and 0.07, respectively. We discuss the reasons for the genetic differences between populations of the Fengshui forests and nature reserves and propose conservation strategies for the Fengshui forest.