Efficacy and Prognosis of Hyperbaric Oxygen as Adjuvant Therapy for Neonatal Hypoxic-Ischemic Encephalopathy: A Meta-Analysis Study.

Background: Preclinical and clinical evidence suggests that hyperbaric oxygen therapy (HBOT) may benefit newborns. The effectiveness of HBOT for neonatal hypoxic-ischemic encephalopathy (HIE) remains controversial. We conducted a meta-analysis to evaluate the efficacy and prognosis of HBOT in neonates with HIE. Methods: A systematic search of eight databases was performed for available articles published between January 1, 2015, and September 30, 2020, to identify randomized controlled clinical trials (RCTs) on HBOT for neonatal HIE. Methodological quality assessment was performed by applying the simple procedure detailed by the Cochrane collaboration. Afterward, quality assessment and data analysis were performed using Revman 5.3 software. STATA 15 software was used to detect publication bias as well as for sensitivity analysis. Results: A total of 46 clinical RCTs were selected for the study and included 4,199 patients with neonatal HIE. The results indicated that HBOT significantly improved the total efficiency (TEF) of treatment for neonatal HIE patients [odds ratio (OR) = 4.61, 95% confidence interval (CI) (3.70, 5.75), P < 0.00001] and reduced the risk of sequelae (OR = 0.23, 95% CI (0.16, 0.33), P < 0.00001) and the neonatal behavioral neurological assessment (NBNA) scores [mean difference (MD) = 4.51, 95%CI (3.83,5.19, P < 0.00001)]. Conclusion: In light of the effectiveness of HBOT neonatal HIE, this meta-analysis suggested that HBOT can be a potential therapy for the treatment of neonatal HIE. Due to the heterogeneity of studies protocol and patient selection being only from China, more research is needed before this therapy can be widely implemented in the clinic. Protocol Registration: PROSPERO (ID: CRD42020210639). Available online at: https://www.crd.york.ac.uk/prospero/display_record.php?ID=CRD42020210639.

[Discussion on automated external defibrillator configuration optimization strategy of rapidly developing city: a case study of Bao'an, Shenzhen].

OBJECTIVE: To explore the automated external defibrillator (AED) configuration optimization strategy in line with the characteristics of the rapidly developing cities by analyzing the actual coverage of AED in Bao'an District based on the real world data of out-of-hospital cardiac arrest (OHCA) in Bao'an District, Shenzhen City. METHODS: The data of cardiac arrest database registered in Bao'an District of Shenzhen City from March 1, 2019 to February 29, 2020 were included in a retrospective observational study. The AED coverage of public and non-public areas was analyzed by calculating the minimum distance between the occurrence place of each OHCA event and the nearest AED. The minimum distance ≤ 100 m was set as AED coverage, and the minimum distance > 100 m was set as non-AED coverage. It was assumed that one AED was configured for each OHCA hotspot area, then the AED coverage changes were analyzed. Based on the actual situation that the AED in schools, governments, sports venues, subways, tourist attractions and parks of public areas in Bao'an District could not be obtained at any time within 24 hours, it was assumed that all AED in the public areas could be obtained at any time within 24 hours, the impact of AED available at any time on AED coverage was analyzed. RESULTS: A total of 525 cases of OHCA were enrolled. The highest incidence of OHCA was found in residential and industrial areas [54.5% (286/525) and 14.3% (75/525), respectively]. There were 252 AED in Bao'an District, Shenzhen, and 115 OHCA events occurred within the coverage area of AED. Even if all AED met the ideal state that could be obtained at any time within 24 hours, the coverage rate was only 21.9% (115/525). The AED coverage rate of the public areas and non-public areas was 31.6% (37/117) and 19.1% (78/408) respectively, with uneven distribution, and the AED coverage rate of non-public areas was low. Assuming that the residential community and industrial zone with more than 2 OHCA cases were respectively equipped with one AED, the coverage rate of AED in the non-public areas increased from 19.1% (78/408) to 28.2% (115/408), basically meeting the requirement that AED could be obtained at any time when OHCA events occurred. Some AED in the public areas of Bao'an District were not available at any time within 24 hours. If the ideal state that all AED in the public area could be obtained at any time within 24 hours could be achieved, the AED coverage rate of all regions increased from 16.8% (88/525) to 21.9% (115/525), the AED coverage rate of the public areas increased from 29.1% (34/117) to 31.6% (37/117), the AED coverage rate of the non-public areas increased from 13.2% (54/408) to 19.1% (78/408). CONCLUSIONS: AED configuration in Bao'an District was unevenly distributed, and the coverage rate of AED in non-public areas was low. The allocation strategy for AED in fast-growing cities like Shenzhen should be as follows: on the premise of ensuring AED availability for 24 hours, priority should be given to covering the number of AED in the non-public areas including residential communities and industrial zones; AED is available in the public areas for 24 hours.

Prenatal diagnosis of Duchenne muscular dystrophy revealed a novel mosaic mutation in Dystrophin gene: a case report.

BACKGROUND: Duchenne muscular dystrophies (DMDs) are X-linked recessive neuromuscular disorders with malfunction or absence of the Dystrophin protein. Precise genetic diagnosis is critical for proper planning of patient care and treatment. In this study, we described a Chinese family with mosaic DMD mutations and discussed the best method for prenatal diagnosis and genetic counseling of X-linked familial disorders. METHODS: We investigated all variants of the whole dystrophin gene using multiple DNA samples isolated from the affected family and identified two variants of the DMD gene in a sick boy and two female carriers by targeted next generation sequencing (TNGS), Sanger sequencing, and haplotype analysis. RESULTS: We identified the hemizygous mutation c.6794delG (p.G2265Efs*6) of DMD in the sick boy, which was inherited from his mother. Unexpectedly, a novel heterozygous mutation c.6796delA (p.I2266Ffs*5) of the same gene, which was considered to be a de novo variant, was detected from his younger sister instead of his mother by Sanger sequencing. However, further NGS analysis of the mother and her amniotic fluid samples revealed that the mother carried a low-level mosaic c.6796delA mutation. CONCLUSIONS: We reported two different mutations of the DMD gene in two siblings, including the novel mutation c.6796delA (p.I2266Ffs*5) inherited from the asymptomatic mosaic-carrier mother. This finding has enriched the knowledge of the pathogenesis of DMD. If no mutation is detected in obligate carriers, the administration of intricate STR/NGS/Sanger analysis will provide new ideas on the prenatal diagnosis of DMD.

[Experience in the construction of social first aid service system: the on-demand training of "Baoan Mode"].

At present there is a lack of standardized first-aid training mode in China. It is an important issue in the construction of social first aid system that how to arrange the first aid training and popularization in a planned and reasonable way according to the limited training resources. Under the guidance of Shenzhen City Baoan District government, the "Baoan Mode" of social emergency grid management has been promoted and achieved good results. The characteristics of "Baoan Mode" emergency system are "government leading, department cooperation, expert guidance, science and technology support, and social participation", and it could effectively shortened the emergency radius. As a result, the "Baoan Mode" was unanimously approved by the emergency medicine, media and government. The public first-aid "on-demand training" is to adopt scientific methods to identify the needs of public first aid training in various regions, and then organize and implement training of first aid knowledge and skills in line with the current situation of local society. The pecific measures include (1) establishing a local cardiac arrest database; (2) analyzing the cardiac arrest database to clarify the target population and content of the training; (3) establishing the local public training registration management system. In China's current national conditions, on-demand training is conducive to the rational use of training resources, a planned and reasonable arrangement of public first-aid training. Based on the experience of "Baoan Mode" initiated by the research team of the authors, this paper discusses the "demand training" theory in social first-aid training, aiming to provide reference for the construction of social first-aid training system in China at present.

[Meta-analysis of the curative effect of low molecular weight heparin on acute respiratory distress syndrome].

OBJECTIVE: To observe the clinical effect of low molecular weight heparin on the curative effect of acute respiratory distress syndrome (ARDS) patients through a systematic evaluation method. METHODS: Databases such as PubMed, Cochrane Library database, Web of Science, CNKI, China Biological Medical Literature Database (CBM), VIP, Wanfang database were searched to retrieve randomized controlled trials (RCTs) on low molecular weight heparin and conventional treatment on the efficacy and prognosis of patients with acute lung injury (ALI)/ARDS from the establishment of the database to September 2020. The conventional treatment group was given conventional treatment, and the low molecular heparin group was given low molecular heparin based on conventional treatment. Outcome indicators were oxygenation index (PaO2/FiO2), acute physiology and chronic health evaluation II (APACHE II), activated partial thromboplastin time (APTT), prothrombin time (PT), platelet count (PLT), interleukin-6 (IL-6) and the mortality at 7 days and 28 days. The literature was screened according to the proposed inclusion and exclusion criteria; the Cochrane collaboration evaluation tool was used to evaluate the quality of literature; Meta analysis used RevMan 5.3 software to perform funnel plot analysis on the outcome indicators of the included ≥ 11 literatures, and to evaluate the publication bias. RESULTS: A total of 12 RCTs and 623 patients were included. The results of Meta analysis showed that PaO2/FiO2 increased in the low molecular weight heparin group compared with the conventional treatment group [mean difference (MD) = 72.08, 95% confidence interval (95%CI) was 56.92-87.24, P < 0.000 01], APACHE II (MD = -4.34, 95%CI was -5.73 to -2.96, P < 0.000 01), IL-6 [standardized mean difference (SMD) = -2.13, 95%CI was -2.71 to -1.56, P < 0.000 01] and 7-day mortality [relative risk (RR) = 0.52, 95%CI was 0.30-0.90, P = 0.02] and 28-day mortality (RR = 0.55, 95%CI was 0.34-0.90, P = 0.02) were significantly reduced, APTT was significantly shortened (MD = -0.88, 95%CI was -1.51 to -0.26, P = 0.006); but PT (MD = -0.44, 95%CI was -1.23 to 0.36, P = 0.28) and PLT (MD = -0.08, 95%CI was -18.81 to 18.65, P = 0.99) between the two groups had no statistically significant differences. A funnel chart analysis of PaO2/FiO2 with 11 included papers showed that there might be publication bias. CONCLUSIONS: Low molecular weight heparin can improve the poor oxygenation of ARDS patients, reduce the mortality of ARDS patients in the acute stage and the inflammatory reaction, but it may increase the bleeding risk of ARDS patients.

Incorporating polyoxometalates and organic ligands to pursue 3d-4f heterometallic clusters: a series of {Cr4Ln4} clusters stabilized by phthalic acid and [SiW12O40]4.

By introduction of trilacunary Keggin-type polyoxometalate to the hydrothermal reaction system of Cr3+, Ln3+ and phthalic acid, a series of novel {Cr4Ln4} heterometallic clusters with the formula Cs2[Cr4Ln4(µ4-O)4(µ3-O)4(C8H4O4)4(H2O)12](H3SiW12O40)Cl·23H2O (1-Ln, Ln = Ce, Pr, Nd) and [Cr4Ln4(µ4-O)4(µ3-O)4(C8H4O4)4(H2O)10](H6SiW12O40)Cl2·18H2O (2-Ln, Ln = Sm, Eu, Gd, Tb, Dy, Ho, Er) have been obtained. Single-crystal structural analyses show that 1-Ln and 2-Ln constitute the first cases of Cr-Ln heterometallic clusters stabilized by inorganic polyoxometalate anions and organic ligands. Optical spectra studies demonstrate that 1-Ln and 2-Ln are narrow-gap semiconductors with band gaps of about 1.5 eV. Magnetic investigation shows that compound 2-Dy is a potential single molecule magnet.

A Series of Banana-Shaped 3d-4f Heterometallic Cluster Substituted Polyoxometalates: Syntheses, Crystal Structures, and Magnetic Properties.

Four seven-nuclearity 3d-4f heterometallic cluster {Fe6LnO28} substituted polyoxometalates (HPz)11K4Fe6Ln(µ3-O)2(B-α-GeW9O34)2(GeW6O26)· xH2O (1-Ln, Pz = piperazine, Ln = Tb, Dy, Ho, Er for x = 27, 25, 25, 24, respectively) have been hydrothermally prepared and structurally characterized by single-crystal X-ray diffraction, powder X-ray diffraction, infrared spectrometry, thermogravimetric analyses, elemental analyses, and electrospray ionization mass spectrometry. Single-crystal X-ray diffraction analyses revealed that 1-Ln contain an unprecedented banana-shaped polyanion constructed from an iron-lanthanide heterometallic {Fe6LnO28} cluster, two trilacunary {B-α-GeW9O34} units, and one hexalacunary {GeW6O26} fragment. The magnetic susceptibility surveying proved the presence of antiferromagnetic coupling in 1-Ln.

Immunocytochemical localization of cholinergic amacrine cells in the bat retina.

The purpose of this study was to localize the cholinergic amacrine cells, one of the key elements of a functional retina, in the retina of a microbat, Rhinolophus ferrumequinum. The presence and localization of choline acetyltransferase-immunoreactive (ChAT-IR) cells in the microbat retina were investigated using immunocytochemistry, confocal microscopy, and quantitative analysis. These ChAT-IR cells were present in the ganglion cell layer (GCL) and inner part of the inner nuclear layer (INL), as previously reported in various animals. However, the bat retina also contained some ChAT-IR cells in the outer part of the INL. The dendrites of these cells extended into the outer plexiform layer, and those of the cells in the inner INL extended within the outer part of the inner plexiform layer (IPL). The dendrites of the ChAT-IR cells in the GCL extended into the middle of the IPL and some fibers ramified up to the outer IPL. The average densities of ChAT-IR cells in the GCL, inner INL, and outer INL were 259±31cells/mm2, 469±48cells/mm2, and 59±8cells/mm2, respectively. The average total density of the ChAT-IR cells was 788±58cells/mm2 (mean±S.D.; n=3; 2799±182 cells/retina). We also found that the cholinergic amacrine cells in the bat retina contained calbindin, one of the calcium-binding proteins, but not calretinin or parvalbumin. As the cholinergic amacrine cells play key roles in the direction selectivity and optokinetic eye reflex in the other mammalian retinas, the present study might provide better information of the cytoarchitecture of bat retina and the basic sources for further physiological studies.

Types and density of calbindin D28k-immunoreactive ganglion cells in mouse retina.

Single-cell injection after immunocytochemistry is a reliable technique for classifying neurons by their morphological structure and their expression of a particular protein. The aim of the present study was to classify the morphological types of calbindin D28k-immunoreactive retinal ganglion cells in the mouse using single-cell injection after immunocytochemistry, to estimate the density of calbindin D28k-immunoreactive retinal ganglion cells in the mouse retina. Calbindin D28k is an important calcium-binding protein that is widely expressed in the central nervous system. Calbindin D28k-immunoreactive retinal ganglion cells were identified by immunocytochemistry and then iontophoretically injected with the lipophilic dye, DiI. Subsequently, the injected cells were imaged by confocal microscopy to classify calbindin D28k-immunoreactive retinal ganglion cells based on their dendritic ramification depth within the inner plexiform layer, field size, and morphology. The cells were heterogeneous in morphology: monostratified or bistratified, with small to large dendritic field size and sparse to dense dendritic arbors. At least 10 different morphological types (CB1-CB10) of calbindin D28k-immunoreactive retinal ganglion cells were found in the mouse retina. The density of each cell type was quite variable (1.98-23.76%). The density of calbindin D28k-immunoreactive cells in the ganglion cell layer of the mouse retina was 562 cells/mm(2), 8.18% of calbindin D28k-immunoreactive cells were axon-less displaced amacrine cells, 91.82% were retinal ganglion cells, and approximately 18.17% of mouse retinal ganglion cells expressed calbindin D28k. The selective expression of calbindin D28k in cells with different morphologies may provide important data for further physiological studies of the mouse retina.

Immunocytochemical localization of the calcium-binding proteins calbindin D28K, calretinin, and parvalbumin in bat visual cortex.

It is a common misconception that bats are blind, and various studies have suggested that bats have visual abilities. The purpose of this study was to investigate the cytoarchitecture of calbindin D28K (CB)-, calretinin (CR)-, and parvalbumin (PV)-immunoreactive (IR) neurons in the bat visual cortex using immunocytochemistry. The highest density of CB- and PV-IR neurons was located in layer IV of the visual cortex. The majority of CB- and PV-IR neurons were characterized by a stellate or round/oval shape. CR-IR neurons were predominantly located in layers II/III, and the cells were principally round/oval in shape. Two-color immunofluorescence revealed that 65.96%, 24.24%, and 77.00% of the CB-, CR-, and PV-IR neurons, respectively, contained gamma-aminobutyric acid (GABA). We observed calcium-binding protein (CBP)-IR neurons in specific layers of the bat visual cortex and in specific cell types. Many of the CBP-IR neurons were GABAergic interneurons. These data provide useful clues to aid in understanding the functional aspects of the bat visual system.

Localization of Nitric Oxide Synthase-containing Neurons in the Bat Visual Cortex and Co-localization with Calcium-binding Proteins.

Microchiroptera (microbats) is a suborder of bats thought to have degenerated vision. However, many recent studies have shown that they have visual ability. In this study, we labeled neuronal nitric oxide synthase (nNOS)-the synthesizing enzyme of the gaseous non-synaptic neurotransmitter nitric oxide-and co-localized it with calbindin D28K (CB), calretinin (CR), and parvalbumin (PV) in the visual cortex of the greater horseshoe bat (Rhinolophus ferrumequinum, a species of microbats). nNOS-immunoreactive (IR) neurons were found in all layers of the visual cortex. Intensely labeled neurons were most common in layer IV, and weakly labeled neurons were most common in layer VI. Majority of the nNOS-IR neurons were round- or oval-type neurons; no pyramidal-type neurons were found. None of these neurons co-localized with CB, CR, or PV. However, the synthesis of nitric oxide in the bat visual cortex by nNOS does not depend on CB, CR, or PV.

Mutagenesis breeding of high echinocandin B producing strain and further titer improvement with culture medium optimization.

A combination of microbial strain improvement and statistical optimization is investigated to maximize echinocandin B (ECB) production from Aspergillus nidulans ZJB-0817. A classical sequential mutagenesis was studied first by using physical (ultraviolet irradiation at 254 nm) and chemical mutagens (lithium chloride and sodium nitrite). Mutant strain ULN-59 exhibited 2.1-fold increase in ECB production to 1583.1 ± 40.9 mg/L when compared with the parent strain (750.8 ± 32.0 mg/L). This is the first report where mutagenesis is applied in Aspergillus to improve ECB production. Further, fractional factorial design and central composite design were adopted to optimize the culture medium for increasing ECB production by the mutant ULN-59. Results indicated that four culture media including peptone, K2HPO4, mannitol and L-ornithine had significant effects on ECB production. The optimized medium provided another 1.4-fold increase in final ECB concentration to 2285.6 ± 35.6 mg/L compared to the original medium. The results of this study indicated the combined application of a classical mutation and medium optimization can improve effectively ECB production from A. nidulans and could be a promising tool to improve other secondary metabolites production by fungal strains.

[Expression of structural protein VP1 of enterovirus 71 in E.coli and preparation of anti-VP1 monoclonal antibodies].

AIM: To express recombinant structural protein VP1 of enterovirus 71 (EV71) in E.coli and prepare anti-VP1 monoclonal antibodies (mAbs). METHODS: With P1 gene as a template, EV71 VP1 gene was amplified by PCR and cloned into the expression vector pET-32a(+). After transformation into E.coli TG1, the positive clones were screened and sequenced. The recombinant plasmid was then transformed into BLgold (DE3) and the recombinant protein in inclusion bodies was induced by IPTG and detected by SDS-PAGE. After the inclusion bodies were solubilized with 6 mol/L guanidine hydrochloride, we purified VP1 protein using Ni-NTA affinity chromatography, and then immunized the mice with it to prepare the mAbs against VP1 protein. RESULTS: Recombinant VP1 protein was expressed in E.coli. We obtained totally 24 VP1 monoclonal hybridoma cell strains, of which one was EV71 positive determined by Western blotting and five were positive for IFA. CONCLUSION: These mAbs are valuable reagents for the development of new vaccines and detection kits for EV71.