Advances in the Treatment of Neuropathic Pain by Sympathetic Regulation.

PURPOSE OF REVIEW: To explore the mechanism and therapeutic effect of sympathetic nerve regulation on neuropathic pain. RECENT FINDINGS: A comprehensive search was conducted in the PubMed and CNKI libraries, using the following keywords: stele ganglion block, neuropathic pain, sympathetic nerve block, sympathetic chemical destruction, and sympathetic radiofrequency thermocoagulation. We selected and critically reviewed research articles published in English that were related to sympathetic modulation in the treatment of neuropathic pain. The collected literature will be classified according to content and reviewed in combination with experimental results and clinical cases. Neuropathic pain was effectively treated with sympathetic regulation technology. Its mechanism includes the inhibition of sympathetic nerve activity, regulation of the inflammatory response, and inhibition of pain transmission, which greatly alleviates neuropathic pain in patients. Stellate ganglion blocks, thoracic and lumbar sympathectomies, chemical destruction, and radiofrequency thermocoagulation have been widely used to treat neuropathic pain. Sympathetic regulation can effectively relieve pain symptoms and improve the patient's quality of life by inhibiting sympathetic nerve activity, reducing the production and release of pain-related mediators, and inhibiting pain transmission. CT-guided radiofrequency thermocoagulation of the thoracic and lumbar sympathetic nerves is effective and durable, with few complications, and is recommended as a treatment for intractable neuropathic pain.

Clinical significance of episcleral venous fluid wave in gonioscopy-assisted transluminal trabeculotomy.

AIM: To evaluate the clinical significance of checking episcleral venous fluid wave (EVFW) during gonioscopy-assisted transluminal trabeculotomy (GATT) in patients with open angle glaucoma (OAG). METHODS: This retrospective case series study comprised 30 patients (45 eyes) with OAG underwent GATT. The location and extent of EVFW were examined and graded after intraoperative compression flushing of the anterior chamber angle during the operation. Patients were followed up for 1y. A complete success for surgery is defined as a postoperative intraocular pressure (IOP) <18 mm Hg without any anti-glaucoma medication. IOP<18 mm Hg with less than two anti-glaucoma medications is defined as qualified success, while the control of IOP requiring three anti-glaucoma medications is considered as unsuccess. RESULTS: The mean IOP was 35.38±7.16 mm Hg before surgery and 15.52±4.22 mm Hg 1y after surgery (P<0.01). The average number of anti-glaucoma medication was 2.8±1.2 (2-4) preoperation and 0.6±1.3 (0-3) 1y postoperation (P<0.01). The success rate of the operation was 93.33%. Complete success rate was 66.67%, qualified success rate was 26.67%, and 6.66% of unsuccessful cases required reoperation. EVFW of all cases was grade 2-4, and the percentages of grade 2, 3 and 4 were 33.33%, 40.0% and 26.67%, respectively. The distribution and percentage of EVFW were inferior (36%), nasal (28%), superior (20%), and temporal (16%). The EVFW grade of complete success patients was 3.4±0.6 (3-4), and that of qualified success patients was 2.6±1.0 (2-4). The larger the range of EVFW, the lower the IOP, and the better the IOP reduction effect. CONCLUSION: During GATT surgery, pressurized irrigation of anterior chamber to check EVFW can reduce the outflow resistance of aqueous humor and increase the effect of postoperative IOP. The range of EVFW is negatively correlated with postoperative IOP. Therefore, EVFW may be a valuable prognostic indicator for the success of GATT surgery.

Arsenic pollution, geochemical fractions, and leaching characteristics in soil samples from four contaminated sites in the Beijiang River Basin, South China.

To remediate historically polluted sites before their land-use changes, it is essential to understand the concentration distribution, geochemical fraction, and migratory behavior of As in soil with varied particle sizes for the use of a sieving procedure. This study investigated the amount and percentage of As in soil with different particle sizes (0.25, 0.25-1, and 1-2 mm) as well as its toxicity characteristic in leaching procedure at four previously contaminated sites in the Beijiang River Basin, South China. The results showed that the total As concentration in the collected soils ranged from 70.1 to 402.8 mg/kg, and only a few percent of soil particle samples had As contents below the local risk threshold value of 60 mg/kg. The amorphous hydrous oxide bound, crystalline hydrous oxide bound, and residual fractions (F3-F5) of the geochemical fraction of As in soil of polluted sites accounted for 82.2-95.7% of the total As distribution. However, the concentration of As in non-specifically bound fractions increased with the mass ratio of soil with coarse particle sizes due to the negative correlation of Fe-bearing minerals concentration with the mass ratio of soil with coarse particle size. According to redundancy analysis, soil with coarse particle sizes and non-specifically bound As were mostly responsible for the As concentration in the leachate. These findings confirmed that a sieving process was not suitable for the remediation of soil As at four historically contaminated sites in the Beijiang River Basin due to the high concentration of As in soil and non-negligible environmental risk of labile extractable As in soil with coarse particle size.

[Chronic Injury of Mice Bone Marrow Multipotent Hematopoietic Progenitor Cells Induced by Ionizing Radiation].

OBJECTIVE: To explore the chronic injury and its possible mechanism of ionizing radiation on multipotent hematopoietic progenitor cells (MPPs) by determining the related indicators of MPPs in bone marrow of mice post-radiation. METHODS: Sixteen C57BL/6 adult mice were randomly divided into normal control and irradiation groups, 8 mice in each group. The mice in irradiation group were exposed to 6 Gy X-ray. The proportion of bone marrow MPPs, their apoptosis and proliferation 2 months after irradiation were detected by flow cytometry. Mitochondrial activity and levels of reactive oxygen species (ROS) in each MPPs population were detected by Mitotracker Red and DCFDA probes, and the senescent state of MPPs in the bone marrow was analyzed. RESULTS: Ionizing radiation could reduce the proportion of MPPs in mouse bone marrow. The proportions and numbers of MPP1, MPP3 and MPP4 in the bone marrow were significantly decreased after whole-body irradiation with 6 Gy X-ray (P<0.05). In addition, radiation significantly reduced the colony-forming capacity of MPPs in bone marrow (P<0.05), the proportions of apoptotic cells in the MPP1 and MPP4 cell populations increased significantly in the bone marrow (P<0.05). The activity of mitochondria was significantly reduced in the bone marrow MPP2, MPP3 and MPP4 cell populations compared with that of the control group (P<0.05). It was also found that the radiation could significantly increase the ROS levels of MPPs in bone marrow, and the content of ROS in the MPP2, MPP3 and MPP4 cell population of the bone marrow was significantly increased(P<0.05). The senescent cells ratios of MPP1, MPP3 and MPP4 cells in the bone marrow after irradiation were significantly higher than those in the control group (P<0.05). CONCLUSION: Ionizing radiation can cause chronic MPPs damage in mice, which is closely associated with persistent oxidative stress, cells apoptosis, and cellular senescence.

Early Diagnosis of Herpes Zoster Neuralgia: A Narrative Review.

BACKGROUND: Early intervention reduces the incidence of postherpetic neuralgia (PHN). Typical shingles are easy to diagnose; however, there is no clear diagnostic method for neuralgia symptoms manifested before the onset of the rash, which can easily cause misdiagnosis. This not only increases the patient's pain, medical expenses, and mental burden, but more importantly, delays the valuable time for early treatment of shingles, and increases the probability of complications and PHN. OBJECTIVE: In this paper, the diagnostic methods of preherpetic neuralgia were summarized and analyzed, and the current challenges were put forward to provide directions for the early diagnosis of herpes zoster (HZ) in the future. METHODS: PubMed, and China National Knowledge Infrastructure (CNKI) libraries were searched using the terms "herpes zoster," "before the blistering," "diagnosis," and "neuralgia." Clinical trials, reviews, and case reports were collected and reviewed. The period of literature search is from 1 January 1980 to 1 October 2022. RESULTS: The early diagnosis of herpes zoster neuralgia can reduce misdiagnosis and mistreatment, and timely and effective intervention can significantly reduce the incidence of PHN. The body may possess a mechanism that limits the local breakthrough of the virus in the skin, causing blistering later than the onset of pain. Changes in the plasma proteins of patients with varicella-zoster virus shingles neuralgia may be used as an early diagnostic indicator in patients with HZ neuralgia before eruption. CONCLUSION: Early diagnosis of HZ neuralgia before eruption can facilitate timely targeted treatment, thereby reducing the incidence of PHN. Proteomic quantitative analysis and validation results can serve as a simple, micro, rapid, and accurate diagnostic method.

Report on Cardiac Gross Pathologic Measurements of Sudden Cardiac Death in Adults.

OBJECTIVES: To analyze the gross pathological data of sudden cardiac death (SCD) with different causes, to provide data support for the identification of sudden cardiac death with unknown causes. METHODS: A total of 167 adult SCD cases in the archive of the Forensic Expertise Institute of Nanjing Medical University from 2010 to 2020 were collected. The gross pathological data of SCD cases were summarized and the characteristics of different causes of death were statistically analyzed. RESULTS: The ratio of male to female SCD cases was 3.4∶1. Coronary heart disease was the leading cause of SCD, and mainly distributed in people over 40 years old. SCD caused by myocarditis was mainly distributed in young people and the mean age of death was (34.00±9.55) years. By analyzing the differences in cardiac pathological parameters of SCD with different causes, it was found that the aortic valve circumference was significantly dilated in the SCD caused by aortic aneurysm or dissection (P<0.05). The heart weight of SCD caused by aortic aneurysm or dissection and combined factors was greater, and both pulmonary and tricuspid valvular rings were dilated in the SCD caused by combined factors in adult males (P<0.05). CONCLUSIONS: Various gross pathological measures of SCD with different causes are different, which has reference value in the cause of death identification of SCD.

Diagnostic value of novel hepatic fibrosis markers in assessing cirrhosis in patients with chronic hepatitis C / 中华肝脏病杂志

Objective: To investigate the efficacy of chitinase-3-like protein 1 (CHI3L1) and Golgi protein 73 (GP73) in the diagnosis of cirrhosis and the dynamic changes of CHI3L1 and GP73 after HCV clearance in patients with chronic hepatitis C (CHC) treated with direct-acting antiviral drugs (DAAs). The comparison of continuous variables of normal distribution were statistically analyzed by ANOVA and t-test. The comparison of continuous variables of non-normal distribution were statistically analyzed by rank sum test. The categorical variables were statistically analyzed by Fisher's exact test and χ(2) test. Correlation analysis was performed using Spearman correlation analysis. Methods: Data of 105 patients with CHC diagnosed from January 2017 to December 2019 were collected. The receiver operating characteristic curve (ROC curve) was plotted to study the efficacy of serum CHI3L1 and GP73 for the diagnosis of cirrhosis. Friedman test was used to compare CHI3L1 and GP73 change characteristics. Results: The areas under the ROC curve for CHI3L1 and GP73 in the diagnosis of cirrhosis at baseline were 0.939 and 0.839, respectively. Serum levels of CHI3L1 and GP73 in the DAAs group decreased significantly at the end of treatment compared with baseline [123.79 (60.25, 178.80) ng/ml vs. 118.20 (47.68, 151.36) ng/ml, P = 0.001; 105.73 (85.05, 130.69) ng/ml vs. 95.52 (69.52, 118.97) ng/ml, P = 0.001]. Serum CHI3L1 and GP73 in the pegylated interferon combined with ribavirin (PR) group were significantly lower at the end of 24 weeks of treatment than the baseline [89.15 (39.15, 149.74) ng/ml vs. 69.98 (20.52, 71.96) ng/ml, P < 0.05; 85.07 (60.07, 121) ng/ml vs. 54.17 (29.17, 78.65) ng/ml, P < 0.05]. Conclusion: CHI3L1 and GP73 are sensitive serological markers that can be used to monitor the fibrosis prognosis in CHC patients during treatment and after obtaining a sustained virological response. Serum CHI3L1 and GP73 levels in the DAAs group decreased earlier than those in the PR group, and the serum CHI3L1 levels in the untreated group increased compared with the baseline at about two years of follow-up.

Size- and density-dependent acoustic differential bioassembly of spatially-defined heterocellular architecture.

Emerging acoustic bioassembly represents an attractive strategy to build cellular closely-packed organotypic constructs in a tunable manner for biofabrication. However, simultaneously assemble heterogeneous cell types into heterocellular functional units with spatially-defined cell arrangements, such as complementary and sandwich cytoarchitectures, remains a long-lasting challenge. To overcome this challenge, herein we present an acoustic differential bioassembly technique to assemble different cell types at the distinct positions of the acoustic field based on their inherent physical characteristics including cellular size and buoyant density. Specifically, different cell types can be differentially assembled beneath the nodal or the antinode regions of the Faraday wave to form complementary cytoarchitectures, or be selectively positioned at the center or edge area beneath either the nodal or the antinode regions to form sandwich cytoarchitectures. Using this technique, we assemble human induced pluripotent stem cell-derived liver spheroids and endothelial cells into hexagonal cytoarchitecturesin vitroto mimic the cord and sinusoid structures in the hepatic lobules. This hepatic lobule model reconstitutes liver metabolic and synthetic functions, such as albumin secretion and urea production. Overall, the acoustic differential bioassembly technique facilitates the construction of human relevantin vitroorganotypic models with spatially-defined heterocellular architectures, and can potentially find wide applications in tissue engineering and regenerative medicine.

Sevoflurane promotes premature differentiation of dopaminergic neurons in hiPSC-derived midbrain organoids.

Background: Conventional animal models used in corresponding basic studies are distinct from humans in terms of the brain's development trajectory, tissue cytoarchitecture and cell types, making it difficult to accurately evaluate the potential adverse effects of anesthetic treatments on human fetal brain development. This study investigated the effects of sevoflurane on the midbrain's development and cytopathology using human physiologically-relevant midbrain organoids. Methods: Monolayer human induced pluripotent stem cells (hiPSC)-derived human floor plate cells and three-dimensional hiPSC-derived midbrain organoids (hMBOs) were exposed to 2% (v/v) sevoflurane for 2 or 6 h, followed by expansion or differentiation culture. Then, immunofluorescence, real-time PCR, EdU assay, Tunnel assay, and transcriptome sequencing were performed to examine the effects of sevoflurane on the midbrain's development. Results: We found that 2% sevoflurane exposure inhibited hFPCs' proliferation (differentiation culture: 7.2% ± 0.3% VS. 13.3% ± 0.7%, p = 0.0043; expansion culture: 48% ± 2.2% VS. 35.2% ± 1.4%, p = 0.0002) and increased their apoptosis, but did not affect their differentiation into human dopaminergic neurons After 6 h, 2% sevoflurane exposure inhibited cell proliferation (62.8% ± 5.6% VS. 100% ± 5.5%, p = 0.0065) and enhanced the premature differentiation of hMBOs (246% ± 5.2% VS. 100% ± 28%, p = 0.0065). The RNA-seq results showed long-term exposure to sevoflurane up regulates some transcription factors in the differentiation of dopaminergic neurons, while short-term exposure to sevoflurane has a weak up-regulation effect on these transcription factors. Conclusion: This study revealed that long-term exposure to sevoflurane could promote the premature differentiation of hMBOs, while short-term exposure had negligible effects, suggesting that long-term exposure to sevoflurane in pregnant women may lead to fetals' midbrain development disorder.

Development of a high-throughput micropatterned agarose scaffold for consistent and reproducible hPSC-derived liver organoids.

Liver organoids represent emerging human-relevantin vitroliver models that have a wide range of biomedical applications in basic medical studies and preclinical drug discovery. However, the generation of liver organoids currently relies on the conventional Matrigel dome method, which lacks precise microenvironmental control over organoid growth and results in significant heterogeneity of the formed liver organoids. Here, we demonstrate a novel high-throughput culture method to generate uniform liver organoids from human pluripotent stem cell-derived foregut stem cells in micropatterned agarose scaffold. By using this approach, more than 8000 uniformly-sized liver organoids containing liver parenchyma cells, non-parenchymal cells, and a unique stem cell niche could be efficiently and reproducibly generated in a 48-well plate with a size coefficient of variation significance smaller than that in the Matrigel dome. Additionally, the liver organoids highly expressed liver-specific markers, including albumin (ALB), hepatocyte nuclear factor 4 alpha (HNF4α), and alpha-fetoprotein (AFP), and displayed liver functions, such as lipid accumulation, glycogen synthesis, ALB secretion, and urea synthesis. As a proof of concept, we evaluated the acute hepatotoxicity of acetaminophen (APAP) in these organoids and observed APAP-induced liver fibrosis. Overall, we expect that the liver organoids will facilitate wide biomedical applications in hepatotoxicity analysis and liver disease modeling.

Ensemble Learning-Based Approach for Gas Detection Using an Electronic Nose in Robotic Applications.

Detecting chemical compounds using electronic noses is important in many gas sensing related applications. A gas detection system is supposed to indicate a significant event, such as the presence of new chemical compounds or a noteworthy change of concentration levels. Existing gas detection methods typically rely on prior knowledge of target analytes to prepare a dedicated, supervised learning model. However, in some scenarios, such as emergency response, not all the analytes of concern are a priori known and their presence are unlikely to be controlled. In this paper, we take a step towards addressing this issue by proposing an ensemble learning based approach (ELBA) that integrates several one-class classifiers and learns online. The proposed approach is initialized by training several one-class models using clean air only. During the sampling process, the initialized system detects the presence of chemicals, allowing to learn another one-class model and update existing models with self-labelled data. We validated the proposed approach with real-world experiments, in which a mobile robot equipped with an e-nose was remotely controlled to interact with different chemical analytes in an uncontrolled environment. We demonstrated that the ELBA algorithm not only can detect gas exposures but also recognize baseline responses under a suspect short-term sensor drift condition. Depending on the problem setups in practical applications, the present work can be easily hybridized to integrate other supervised learning models when the prior knowledge of target analytes is partially available.

Anemoside B4 inhibits enterovirus 71 propagation in mice through upregulating 14-3-3 expression and type I interferon responses.

Enterovirus 71 (EV71) is the major pathogens of human hand, foot, and mouth disease (HFMD). EV71 efficiently escapes innate immunity responses of the host to cause infection. At present, no effective antiviral drugs for EV71 are available. Anemoside B4 (B4) is a natural saponin isolated from the roots of Pulsatilla chinensis (Bunge) Regel. P. chinensis extracts that shows a wide variety of biological activities. In this study, we investigated the antiviral activities of B4 against EV71 both in cell culture and in suckling mice. We showed that B4 (12.5-200 µM) dose dependently increased the viability of EV71-infected RD cells with an IC50 value of 24.95 ± 0.05 µM against EV71. The antiviral activity of B4 was associated with enhanced interferon (IFN)-ß response, since knockdown of IFN-ß abolished its antiviral activity. We also confirmed that the enhanced IFN response was mediated via activation of retinoic acid-inducible gene I (RIG-I) like receptors (RLRs) pathway, and it was executed by upregulation of 14-3-3 protein, which disrupted the interaction between yes-associated protein (YAP) and interferon regulatory factor 3 (IRF3). By using amino acids in cell culture (SILAC)-based proteomics profiling, we identified the Hippo pathway as the top-ranking functional cluster in B4-treated EV71-infected cells. In vivo experiments were conducted in suckling mice (2-day-old) infected with EV71 and subsequently B4 (200 mg · kg-1 · d-1, i.p.) was administered for 16 days. We showed that B4 administration effectively suppressed EV71 replication and improved muscle inflammation and limb activity. Meanwhile, B4 administration regulated the expressions of HFMD biomarkers IL-10 and IFN-γ, attenuating complications of EV71 infection. Collectively, our results suggest that B4 could enhance the antiviral effect of IFN-ß by orchestrating Hippo and RLRs pathway, and B4 would be a potential lead compound for developing an anti-EV71 drug.

Effect of Huazhuo Jiedu Huoxue Tongluo Prescription on Brain-gut Axis of Rats with Cerebral Ischemia-reperfusion Injury Based on 16S rRNA Sequencing of Intestinal Flora / 中国实验方剂学杂志

ObjectiveTo study the effect of Huazhuo Jiedu Huoxue Tongluo （HJHT） prescription on the intestinal flora in rats with cerebral ischemia-reperfusion injury， and to explore the mechanism of Chinese medicinal prescription regulating intestinal flora to restore the balance of brain-gut axis. MethodFifty male SPF SD rats were randomly assigned into sham group， model group， high-dose HJHT group （25.0 g·kg-1）， medium-dose HJHT group （12.5 g·kg-1）， and low-dose HJHT group （6.25 g·kg-1）， with 10 rats in each group. The rat model of permanent middle cerebral artery infarction was established according to Longa method and previous research experience， and reperfusion was performed 2 h after ischemia. The recovery of neurological function deficit and the percentage of cerebral infarction area were detected 72 h after administration. Real-time PCR was performed to detect the mRNA levels of Occludin and zonula occludens-1 （ZO-1） in rat colon. Hematoxylin-eosin （HE） staining was conducted to reveal the intestinal damage. The feces of 6 rats in each group were collected for 16S rRNA sequencing. The expression of Treg and Th17 in intestinal tissue， peripheral blood， and brain tissue were detected. ResultCompared with the sham group， the model group showed obvious neurological deficit （P<0.05） and large cerebral infarction area （P<0.05）. High-dose and medium-doses HJHT alleviated the symptoms of neurological impairment （P<0.05） and reduce the cerebral infarction area （P<0.05） compared with the model group. Compared with the sham group， the model group showed destroyed structure of colonic mucosa and incomplete epithelial cells and goblet cells， while high-dose and medium-doses HJHT alleviated such changes. The mRNA levels of Occludin and ZO-1 in the model group were lower than those in the sham group （P<0.05），and the high-dose HJHT groups were higher than the model group （P<0.05）. The intestinal flora structure was significantly different between the model group and the sham group while similar between the high-dose HJHT group and sham group. Compared with the sham group， the model group showed down-regulated expression of Treg and up-regulated expression of Th17 in the intestinal tissue， peripheral blood， and brain tissue， and high-dose and medium-dose HJHT alleviated the changes in the expression of Treg and Th17 in the model group （P<0.05）. ConclusionHuazhuo Jiedu Huoxue Tongluo prescription may improve the permeability of intestinal wall by adjusting the abundance and diversity of intestinal microorganisms to reduce the migration of intestinal Th17 cells toward the ischemic lateral brain tissue， mitigate the inflammatory response， and thus alleviate the cerebral ischemia-reperfusion injury in rats.

Changes of antithrombotic prescription in atrial fibrillation patients with acute coronary syndrome or percutaneous coronary intervention and the subsequent impact on long-term outcomes: a longitudinal cohort study.

OBJECTIVES: The choice of optimal antithrombotic therapy in atrial fibrillation (AF) patients with acute coronary syndrome (ACS) or percutaneous coronary intervention (PCI) remains controversial. The aim of this longitudinal cohort study is to investigate the prescribing pattern of antithrombotic regimen in different cohorts and its subsequent impact. SETTING AND DESIGN: Longitudinal data from the Tri-Service General Hospital-Coronary Heart Disease (TSGH-CHD) registry, between January 2016 and August 2018 was screened. PARTICIPANTS AND METHOD: Patients with prior history of nonvalvular AF, who had ACS presentation or underwent PCI were selected, and these patients were divided into cohort 1 and cohort 2, according to the index date of antithrombotic prescription before and after the PIONEER AF-PCI study. PRIMARY AND SECONDARY OUTCOMES: The primary safety endpoints were composites of major bleeding and/or clinically relevant non-major bleeding. The secondary efficacy endpoints included the occurrence of all-cause mortality, stroke/systemic embolization, nonfatal myocardial infarction (MI), and >30-days coronary revascularization. RESULTS: A total of 121 patients were included into analysis (cohort 1=35; cohort 2=86). Comparing with cohort 1, the prescription rate of triple antithrombotic therapy (TAT) increased from 17.1 to 38.4%, especially the regimen with dual antiplatelet therapy (DAPT) plus low-dose non-vitamin-K dependent oral anticoagulation (NOAC). However, the prescription rate of dual antithrombotic therapy (DAT) decreased (14.3-10.5%), as well as the prescription rate of DAPT (68.6-51.2%). These changes of antithrombotic prescription across different cohorts were not associated with risk of adverse safety (HR= 0.87; 95% CI, 0.42-1.80, p=0.710) and efficacy outcomes (HR=0.96; 95% CI, 0.40-2.32, p=0.930). CONCLUSIONS: Entering the NOAC era, the prescription of TAT increased alongside the decrease in DAT. As the prescription rate of DAPT without anticoagulation remained high, future efforts are mandatory to improve the implementation of guidelines and clinical practice.

Complete mitochondrial DNA sequence of yellow-legged buttonquail (Turnix tanki).

The Yellow-legged Buttonquail Turnix tanki is a species of the genus Turnix, which belongs to the order Charadriiformes. It is distributed across almost all of China. The International Union for Conservation of Nature has assessed the bird's conservation status as 'Least Concern (LC).' We sequenced the complete mitogenome of T. tanki and examined its phylogenetic relationship with other charadriiformes species. The mitochondrial DNA is packaged in a compact 17,620 base pair circular molecule with A + T content of 57.90%. It contains 37 typical mitochondrial genes, including 13 protein-coding genes, two rRNAs and 22 tRNAs, and two non-coding regions. We reconstructed a phylogenetic tree based on mitogenome sequences of five Turnicidae species and one outgroup. Phylogenetic analysis indicated that T. tanki is a sister to T. suscitator.

Temporal-spatial variation and the affecting factors of protected areas in Guizhou, China.

The establishment of protected areas is the bottom line of ecological security for promoting the construction of ecological civilization and supporting economic and social development, which is an important strategy to realize sustainable development and maintain ecological security. In order to reveal the large spatial process of protected areas and its influencing factors, we used the methods of nearest neighbor index, kernel density, and standard deviational ellipse to analyze the temporal-spatial variation characteristics of the protected areas in Guizhou Province from 2002 to 2017, as well as the influencing factors combined with geo-detectors. The results showed that, during the study period, the number, area, and types of protected areas in Guizhou Province showed a diversified and rapid development, forming a protected area system with nature reserves, forest parks and scenic spots as the main body and wetland parks, geoparks and natural heritage sites as the supplement. The spatial cohesion of protected areas was strengthened, the scope of spatial distribution was expanding, and the speed of spatial movement was declining, forming a spatial pattern dominated by the northeast-southwest direction and gradually stable. The coalescence process in protected areas was strongly influenced by topography and vegetation distribution. The protected areas tended to cluster in gentle terrain around rivers and mountains and in areas of concentrated vegetation. The spatial differentiation of protected areas was jointly affected by multiple factors at different levels. The explanatory power of different factors to the spatial differentiation of protected areas was different. Among them, the normalized difference vegetation index, areas of forest and highway mileage were the common main factors affecting the spatial differentiation of the number and area of protected areas, and the explanatory power of different factors was significantly consolidated after interaction, characterized as nonlinear or bi-factor enhancement.

Increasing Long-Chain Dicarboxylic Acid Production in Candida tropicalis by Engineering Fatty Transporters.

Candida tropicalis can metabolize alkanes or fatty acids to produce long-chain dicarboxylic acids (DCAs). Fatty acid transporters located on the cell or peroxisome membrane may play an important role in this process. Using amino acid sequence homologous alignment, two putative proteins, CtFat1p and CtPxa1p, located on the cell and peroxisome membrane were found, respectively. Moreover, single- and double-knockout homologous recombination technology was used to study ctfat1p and ctpxa1p gene effects on DCA synthesis. In comparison to the wild-type strain, long-chain DCA yield decreased by 65.14%, 88.38% and 56.19% after single and double-copy knockout of ctfat1p genes and double-copy knockout of ctpxa1p genes, respectively, indicating that the knockout of ctfat1p and ctpxa1p genes had a significant effect on the conversion of oils and fats into long-chain DCAs by C. tropicalis. However, the yield of long-chain DCAs increased by 21.90% after single-knockout of the ctpxa1p gene, indicating that the single-knockout of the ctpxa1p gene may reduce fatty acid transport to peroxisome for further oxidation. Moreover, to improve the intracellular transport rate of fatty acids, ctfat1p copy number increased, increasing DCA yield by 30.10%. These results may provide useful information for enhancing the production of long-chain DCAs by C. tropicalis.

Increasing L-lysine production in Corynebacterium glutamicum by engineering amino acid transporters.

Corynebacterium glutamicum has a long and successful history in the biotechnological production of L-lysine. Besides the adjustment of metabolic pathways, intracellular and extracellular transport systems are critical for the cellular metabolism of L-lysine or its by-products. Here, three amino acid transmembrane transporters, namely, GluE, BrnE/BrnF, and LysP, which are widely present in C. glutamicum strains, were each investigated by gene knockout. In comparison with that in the wild-type strain, the yield of L-lysine increased by 9.0%, 12.3%, and 10.0% after the deletion of the gluE, brnE/brnF, and lysP genes, respectively, in C. glutamicum 23,604. Moreover, the amount of by-product amino acids decreased significantly when the gluE and brnE/brnF genes were deleted. It was also demonstrated that there was no effect on the growth of the strain when the gluE or lysP gene was deleted, whereas the biomass of C. glutamicum WL1702 (ΔbrnE/ΔbrnF) in the fermentation medium was significantly reduced in comparison with that of the wild type. These results also provide useful information for enhancing the production of L-lysine or other amino acids by C. glutamicum.

Enhancement of Production of D-Glucosamine in Escherichia coli by Blocking Three Pathways Involved in the Consumption of GlcN and GlcNAc.

D-Glucosamine is a commonly used dietary supplement that promotes cartilage health in humans. Metabolic flux analysis showed that D-glucosamine production could be increased by blocking three pathways involved in the consumption of glucosamine-6-phosphate and acetylglucosamine-6-phosphate. By homologous single-exchange, two key genes (nanE and murQ) of Escherichia coli BL21 were knocked out, respectively. The D-glucosamine yields of the engineered strains E. coli BL21ΔmurQ and E. coli BL21ΔnanE represented increases by factors of 2.14 and 1.79, respectively. Meanwhile, for bifunctional gene glmU, we only knocked out its glucosamine-1-phosphate acetyltransferase domain by 3D structural analysis to keep the engineered strain E. coli BL21glmU-Δgpa survival, which resulted in an increase in the production of D-glucosamine by a factor of 2.16. Moreover, for further increasing D-glucosamine production, two genes encoding rate-limiting enzymes, named glmS and gna1, were coexpressed by an RBS sequence in those engineered strains. The total concentrations of D-glucosamine in E. coli BL21 glmU-Δgpa', E. coli BL21ΔmurQ', and E. coli BL21ΔnanE' were 2.65 g/L, 1.73 g/L, and 1.38 g/L, which represented increases by factors of 8.83, 5.76, and 3.3, respectively.

Emerging organoid models: leaping forward in cancer research.

Cancer heterogeneity is regarded as the main reason for the failure of conventional cancer therapy. The ability to reconstruct intra- and interpatient heterogeneity in cancer models is crucial for understanding cancer biology as well as for developing personalized anti-cancer therapy. Cancer organoids represent an emerging approach for creating patient-derived in vitro cancer models that closely recapitulate the pathophysiological features of natural tumorigenesis and metastasis. Meanwhile, cancer organoids have recently been utilized in the discovery of personalized anti-cancer therapy and prognostic biomarkers. Further, the synergistic combination of cancer organoids with organ-on-a-chip and 3D bioprinting presents a new avenue in the development of more sophisticated and optimized model systems to recapitulate complex cancer-stroma or multiorgan metastasis. Here, we summarize the recent advances in cancer organoids from a perspective of the in vitro emulation of natural cancer evolution and the applications in personalized cancer theranostics. We also discuss the challenges and trends in reconstructing more comprehensive cancer models for basic and clinical cancer research.