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Icaritin is a prenylflavonoid present in the Chinese herbal medicinal plant Epimedium spp. and is under investigation in a phase III clinical trial for advanced hepatocellular carcinoma. Here, we report the biosynthesis of icaritin from glucose by engineered microbial strains. We initially designed an artificial icaritin biosynthetic pathway by identifying a novel prenyltransferase from the Berberidaceae-family species Epimedium sagittatum (EsPT2) that catalyzes the C8 prenylation of kaempferol to yield 8-prenlykaempferol and a novel methyltransferase GmOMT2 from soybean to transfer a methyl to C4’-OH of 8-prenlykaempferol to produce icaritin. We next introduced 11 heterologous genes and modified 12 native yeast genes to construct a yeast strain capable of producing 8-prenylkaempferol with high efficiency. GmOMT2 was sensitive to low pH and lost its activity when expressed in the yeast cytoplasm. By relocating GmOMT2 into mitochondria (higher pH than cytoplasm) of the 8-prenylkaempferol–producing yeast strain or co-culturing the 8-prenylkaempferol–producing yeast with an Escherichia coli strain expressing GmOMT2, we obtained icaritin yields of 7.2 and 19.7 mg/L, respectively. Beyond the characterizing two previously unknown plant enzymes and conducting the first biosynthesis of icaritin from glucose, we describe two strategies of overcoming the widespread issue of incompatible pH conditions encountered in basic and applied bioproduction research. Our findings will facilitate industrial-scale production of icaritin and other prenylflavonoids.

Non-suicidal self-harm (NSSH) and chronic insufficient sleep are both major health problems during the transition from childhood to adolescence. We examined to identify sleep duration trajectories from childhood to adolescence and their associations with subsequent risk of NSSH. A cohort of children around the period of pubertal onset (7–9 years old) were followed from 2013 over 6 years. Group-based trajectory modeling was recruited to identify sleep duration trajectories derived from 5 repeated measures. Association between sleep duration trajectories with the risk of NSSH was examined using multivariate logistic regression model. Nonlinear dose–response associations between sleep duration and NSSH risk were also assessed using restricted cubic spline models. Of the 1973 participants included in the study (mean ± SD, 8.1 ± 0.9 years age at baseline, 41.1% female). Three sleep duration trajectories were identified: persistent sleeping ≥ 8 h/day (27.7%), moderately decreasing (60.8%) and rapidly decreasing (11.5%) sleep duration groups. After multivariable adjustment for covariates, compared with the persistent sleeping ≥ 8 h/day group, the odds ratio of NSSH was 2.58 (95% CI 1.92, 3.45) for the moderately decreasing group, and 4.16 (2.86, 6.04) for rapidly decreasing group. In dose–response analysis, sleep duration was associated with NSSH risk in a non-linear fashion (χ2 = 25.16, Pnonlinearity

Background In this study we compared intraoperative bleeding and regional cerebral oxygenation in patients with different degrees of the reverse Trendelenburg position (RTP) during endoscopic sinus surgery (ESS). Methods In total, 120 patients with chronic rhinosinusitis treated by ESS were randomly assigned to the following 4 groups: a horizontal position (HP) group, and 5°, 10°, and 15° RTP (5-RTP, 10-RTP, and 15-RTP, respectively) groups. The primary outcome was the Boezaart grading scale (BS). The cerebral oxygen saturation (ScO2 ), total blood loss, numerical rating scale (NRS) scores, and complications were also recorded. Results The median BS values in the HP, 5-RTP, 10-RTP, and 15-RTP groups were 2.0, 2.0, 2.1, and 1.7, respectively. Multiple pairwise comparisons of the BS showed significant differences between the 15-RTP group and the other 3 groups (HP, 5-RTP, and 10-RTP). Regarding the NRS and bleeding rate, significant differences were found between the HP and 15-RTP groups. No difference was found in ScO2 among the 4 groups, and no cerebral desaturation events occurred in any group. No complications, including vital organ (heart, brain, and kidney) dysfunction problems, were reported in this study during hospitalization. Conclusion Compared with HP, 5-RTP, and 10-RTP, 15-RTP can improve visual clarity during ESS, and ScO2 is not affected by the degree of RTP. No cerebral deoxygenation or vital organ dysfunction was observed in this study. Therefore, we recommend 15-RTP with moderate deliberate hypotension for ESS.

Cancer is one of the most serious diseases that are harmful to human health. Systemic chemotherapy is an optimal therapeutic strategy for the treatment of cancer, but great difficulty has been encountered in its administration in the form of multidrug resistance (MDR). As an enzyme on the outer cell surface, CD13 is documented to be involved in the MDR development of tumor cells. In this review, we will focus on the role of CD13 in MDR generation based on the current evidence.

Introduction Although weight loss is common in Parkinson's disease (PD), longitudinal studies assessing weight and body composition changes are limited. Methods In this three-year longitudinal study, 125 subjects (77 PD patients and 48 spousal/sibling controls) underwent clinical, biochemical and body composition assessments using dual-energy X-ray absorptiometry. Results Patients were older than controls (65.6 ± 8.9 vs. 62.6 ± 7.1, P = 0.049), with no significant differences in gender, comorbidities, dietary intake and physical activity. Clinically significant weight loss (≥5% from baseline weight) was recorded in 41.6% of patients, with a doubling of cases (6.5 to 13.0%) classified as underweight at study end. Over three years, patients demonstrated greater reductions in BMI (mean −1.2 kg/m2, 95%CI-2.0 to −0.4), whole-body fat percentage (−2.5% points, 95%CI-3.9 to −1.0), fat mass index (FMI) (−0.9 kg/m2, 95%CI-1.4 to −0.4), visceral fat mass (−0.1 kg, 95%CI-0.2 to 0.0), and subcutaneous fat mass (−1.9 kg, 95%CI-3.4 to −0.5) than in controls, with significant group-by-time interactions after adjusting for age and gender. Notably, 31.2% and 53.3% of patients had FMI Conclusions We found progressive loss of weight in PD patients, with greater loss of both visceral and subcutaneous fat, but not muscle, compared to controls. Several associated factors (motor and non-motor disease features) were identified for these changes, providing insights on possible mechanisms and therapeutic targets.

Little is known regarding the educational needs and perspectives of people living with Parkinson’s disease (PD), particularly in Asia. To assess knowledge and perceptions regarding PD in a large multiethnic urban Asian cohort of patients and caregivers. We conducted a survey at a university hospital neurology clinic, using a novel Knowledge and Perception of Parkinson’s Disease Questionnaire (KPPDQ). The KPPDQ had satisfactory psychometric properties among patients and caregivers. Five hundred subjects were recruited with a 97% response rate (211 patients, 273 caregivers). Non-motor symptoms such as urinary problems, visual hallucinations and pain were relatively poorly recognized. Many (≈ 50–80%) respondents incorrectly believed that all PD patients experience tremor, that PD is usually familial, and that there is a cure for PD. About one-half perceived PD to be caused by something the patient had done in the past, and that PD medications were likely to cause internal organ damage. Issues of stigma/shame were relevant to one-third of patients, and 70% of patients perceived themselves to be a burden to others. Two-thirds of participants felt that PD imposed a heavy financial toll. Participants were about equally divided as to whether they would consider treatment with deep brain stimulation, tube feeding or invasive ventilation. Over three-quarters of patients expressed a preference to die at home. Important knowledge gaps, misperceptions and perspectives on PD were identified, highlighting the need for further efforts to raise awareness and provide accurate information regarding PD, and to address patient’s and caregivers’ needs and preferences.

As the outermost hydrophobic layer, cuticular waxes serve as an essential waterproof barrier to protect plants from desiccation, but the mechanism of wax accumulation still remains unclear. We analyzed the response of cuticular wax composition and deposition to drought in three different watermelon germplasms, namely, M20, M08, and J5F, which showed similar stomatal response, but high, moderate, and low tolerance to drought, respectively. Among the identified 28 compounds of cuticular waxes on leaves, more alkanes with chain lengths of C29 and C31 were induced in M20, accompanied by an increased transcript levels of CER1 (very-long-chain aldehyde decarbonylase 1), when compared to that in M08 and J5F. M20 showed higher total wax amount but fewer platelet-like wax crystals on the upper epidermis of leaves under drought, suggesting the prevalence of more intracuticular waxes embedded into the cutin matrix as fillers. These distinct responses of cuticular waxes in M20 conferred low water loss and high tolerance to drought. Melatonin and abscisic acid (ABA), which can be induced by drought, promoted the biosynthesis of alkanes (C29 and C31) but inhibited the accumulation of wax crystals under drought. Moreover, melatonin inhibited the elevation of ABA levels under mild drought but promoted the ABA accumulation under severe drought, indicating that melatonin and ABA function synergistically to regulate wax compositions to limit non-stomatal water loss under severe but not mild drought in watermelon. These findings can be exploited to improve crop tolerance to drought in arid regions.

Cylindrical Li-ion batteries (cells) typically have safety vents in the positive terminal to enable the release of gases that build up inside the battery and thus help reduce the effects of thermal runaway, including fire and explosion. However, the vents are not always effective, and it is critical to understand why. This paper overviews various vent designs and presents two case studies of vents in cylindrical Li-ion batteries that failed to operate properly. The first case study concerns a Sony VCT5 18650 Li-ion battery in which thermal runaway caused the cylindrical casing (can) to rupture. The second case study involves an Ampking 20700 Li-ion battery in which thermal runaway caused side-wall rupture of the casing. In both cases, the vents did not function as intended. Computed tomography (CT) scanning was used to investigate the internal structure of the battery to assess the failure mechanisms of the failed vents. These cases raise concerns about the efficiency, reliability, and safety of some current vent designs.

Background This study was to evaluate the diagnostic significance of Magnetic Resonance Imaging (MRI) in distinguishing temporomandibular disorders (TMD). Methods A total of 684 patients with TMD were included in the study. Diagnosis for TMD was conducted according to the international criteria. Two professional radiologists were selected for professional training, and the Kappa values were compared for the diagnosis results to determine the consistency of the diagnosis. Then MRI images of these 684 patients were analyzed and the diagnosis results were obtained. Results MRI can be used for the diagnosis of TMD. There were significantly more females (518 cases) than males (166 cases) with TMD; Disc displacement with/without reduction is more common in the youth group, with the majority aged 20–30 years. The highest incidence of temporomandibular joint osteoarthrosis is in the 60-year-old age group, followed by the 70-year-old age group. Conclusions Bilateral temporomandibular joint MRI can clearly show their changes; there are significantly more female with TMD than male; osteoarthritis has a significant correlation with age.

Coupling reagents play crucial roles in the iterative construction of amide bonds for the synthesis of peptides and peptide-based derivatives. The novel DIC/Oxyma condensation system featured with the low risk of explosion displayed remarkable abilities to inhibit racemization, along with efficient coupling efficiency in both manual and automated syntheses. Nevertheless, an ideal reaction molar ratio in DIC/Oxyma condensation system and the moderate reaction temperature by manual synthesis remain to be further investigated. Herein, the synthetic efficiencies of different reaction ratios between DIC and Oxyma under moderate reaction temperature were systematically evaluated. The robustness and efficiency of DIC/Oxyma condensation system are validated by the rapid synthesis of linear centipede toxin RhTx. Different folding strategies were applied for the construction of disulfide bridges in RhTx, which was further confirmed in assays of circular dichroism and patch-clamp electrophysiology evaluation. This work establishes the DIC/Oxyma-based accelerated synthesis of peptides under moderate condensation conditions, which is especially useful for the manual synthesis of peptides. Besides, the strategy presented here provides robust technical supports for the large-scale synthesis and oxidative folding of RhTx.

Shen Gui capsule (SGC) has been demonstrated to have a significant treatment effect for coronary heart disease (CHD). Nevertheless, the holistic therapeutic mechanism of SGC in vivo remain poorly interpreted. We aimed to systematically explore the preventive effect and mechanism of SGC on CHD rats using plasma metabolomics strategy. Rat CHD model was established by left anterior descending coronary artery ligation (LAD). Echocardiography, histological analyses of the myocardium and biochemical assays on serum were used to confirm the successful establishment of the CHD model and therapeutic effects of SGC. Then, UHPLC-MS/MS-based plasma metabolomics was combined with multivariate data analysis to screen potential pharmaco biomarkers associated with SGC treatment in the LAD-induced rat CHD model. After SGC treatment, 12 abnormal metabolites considered as potiential pharmaco biomarkers recovered to near normal levels. These biomarkers were involved in several metabolic pathways, including fat and protein metabolism, phenylalanine metabolism, neuroactive ligand-receptor interaction, androgen receptor signaling pathway, and estrone metabolism.These results suggested that SGC achieves therapeutic action on CHD via regulating various aspects of the body such as energy metabolism, neurological disturbances and inflammation, and thus plays a significant role in the treatment of CHD and its complications. The study is useful to systematically understand and analyze the mechanism of SGC's "multipie pathways, multiple levels, multiple targets" prevention and treatment of CHD.

Toll-like receptor 3 (TLR3) is a sensor of endogenous cell necrosis during the process of acute inflammation. Interleukin (IL)-1 receptor antagonist (IL-1Ra) is an anti-inflammatory cytokine and can negatively regulate the pathogenesis of inflammation. However, whether and how activation of TLR3 can regulate IL-1Ra expression has not been clarified. Here, we show that poly(I:C) induces IL-1Ra expression in primarily cultured human fibroblast-like synoviocytes and other types of cells. Induction of IL-1Ra by poly(I:C) was dependent on TLR3, but was independent of melanoma differentiation-­associated protein 5 or retinoic acid-inducible gene I. Interferon regulatory factor 3 (IRF3) directly binds to the IL-1Ra promoter and promotes IL-1Ra expression in response to poly(I:C) stimulation. Induction of IL-1Ra by poly(I:C) was abolished by the inhibition of the NF-κB signaling, attenuated by the inhibition of the PI3K-Akt signaling, enhanced by inhibition of the ERK1/2 or MSK1/2 activation, but was independent of the p38 MAPK signaling. Treatment with poly(I:C) or Sendai virus elevated the levels of serum IL-1Ra in wild-type, but not in TLR3–/– or IRF3–/– mice. Our findings may provide new insights into the intrinsic anti-inflammatory function of TLR3 and double-stranded RNA-induced IL-Ra expression by TLR3 and its regulation.

Although multidrug therapy (MDT) has been widely used for the treatment of leprosy for nearly 40 y, the disease remains a public health concern in some areas. The early detection of leprosy cases is vital to interrupt Mycobacterium leprae transmission, but currently diagnosis is typically achieved during the recognition of clinical symptoms by professional staff performing physical examinations in conjunction with microbiological assessment of slit skin smears (SSSs) and histopathology. In the last 10 y, serum antibody detection tests have emerged to aid leprosy diagnosis. Here we evaluated the ability of antigens NDO-BSA and LID-1 (ML0405 and ML2331) and the conjugate of these, NDO-LID, to detect antibodies in the sera of 113 leprosy patients and 166 control individuals in Yunnan province in southwest China. We found that each antigen was readily detected by sera from multibacillary (MB) patients, with sensitivities of 97.3%, 97.3% and 98.6% for NDO-BSA, LID-1 and NDO-LID, respectively. Even among paucibacillary (PB) patients the antigens detected antibodies in 74.4%, 56.4% and 69.2% of serum samples, respectively. Receiver operating characteristics (ROC) curve analysis indicated that, irrespective of the leprosy case classification as MB or PB, the detection efficiency obtained with NDO-LID was better than that obtained with the other two antigens (with LID-1 being a slightly better than NDO-BSA). Our results indicate the utility of NDO-LID in assisting in the diagnosis of PB and MB leprosy patients and that these antibody detection assays represent powerful diagnostic tools. We suggest that could be implemented into the procedures of local health centres in leprosy-endemic regions to assist in earlier diagnosis.

Background The ability to digest dietary lactose is associated with lactase persistence (LP) in the intestinal lumen in human. The genetic basis of LP has been investigated in many populations in the world. Iran has a long history of pastoralism and the daily consumption of dairy products; thus, we aim to assess how LP has evolved in the Iranian population. We recruited 400 adult individuals from seven Iranian ethnic groups, from whom we investigated their lactose tolerance and screened the genetic variants in their lactase gene locus. Results The LP frequency distribution ranged from 0 to 29.9% in the seven Iranian ethnic groups with an average value of 9.8%. The variants, − 13910*T and − 22018*A, were significantly associated with LP phenotype in Iranians. We found no evidence of hard selective sweep for − 13910*T and − 22018*A in Persians, the largest ethnic group of Iran. The extremely low frequency of − 13915*G in the Iranian population challenged the view that LP distribution in Iran resulted from the demic diffusion, especially mediated by the spread of Islam, from the Arabian Peninsula. Conclusions Our results indicate the distribution of LP in seven ethnic groups across the Iranian plateau. Soft selective sweep rather than hard selective sweep played a substantial role in the evolution of LP in Iranian populations.

Objective To measure blood and follicular antimullerian hormone (AMH) levels in women with polycystic ovary syndrome (PCOS) undergoing assisted reproductive technologies (ART) and to examine the direct action of insulin on AMH expression in human granulosa cells. Design Prospective clinical and experimental study. Setting University Hospital-based laboratory. Patient(s) Women with (n = 86) and without (n = 172) PCOS in ART. Intervention(s) Blood, follicular fluid, and luteinized granulosa cells were collected from PCOS and non-PCOS women in ART. Main Outcome Measure(s) Hormone levels in blood and fluid, and gene expression in granulosa cells. Result(s) Serum levels of AMH were elevated and inversely correlated with embryo cleavage rate in PCOS women in ART. Significant higher levels of AMH were also found in small and large follicles collected from PCOS women compared with non-PCOS women. Luteinized granulosa cells from PCOS women showed higher expression of AMH and its receptor AMHR2. Direct effect of insulin in increasing the expression of AMH in the isolated luteinized granulosa cells was observed, with the PCOS granulosa cells responding to a high dose of insulin. Cotreatment with AMH attenuated insulin-induced aromatase expression in the luteinized granulosa cells. Conclusion(s) These results suggest that insulin may contribute to AMH elevation in PCOS and that AMH counteracts insulin-promoted aromatase expression in granulosa cells.

Hepatitis B virus (HBV) has a crucial role in the progression of hepatocellular carcinoma (HCC). Tumour cells must develop anoikis resistance in order to survive before metastasis. This study aimed to investigate the mechanism of IQGAP1 in HBV-mediated anoikis evasion and metastasis in HCC cells. IQGAP1 expression was detected by immunohistochemistry, real-time PCR and immunoblot analysis. Lentiviral-mediated stable upregulation or knockdown of IGAQP1, immunoprecipitation, etc. were used in function and mechanism study. IQGAP1 was markedly upregulated in HBV-positive compared with HBV-negative HCC cells and tissues. IQGAP1 was positively correlated to poor prognosis of HBV-associated HCC patients. IQGAP1 overexpression significantly enhanced the anchorage-independent growth and metastasis, whereas IQGAP1-deficient HCC cells are more sensitive to anoikis. Mechanistically, we found that HBV-induced ROS enhanced the association of IQGAP1 and Rac1 that activated Rac1, leading to phosphorylation of Src/FAK pathway. Antioxidants efficiently inhibited IQGAP1-mediated anoikis resistance and metastasis. Our study indicated an important mechanism by which upregulated IQGAP1 by HBV promoted anoikis resistance, migration and invasion of HCC cells through Rac1-dependent ROS accumulation and activation of Src/FAK signalling, suggesting IQGAP1 as a prognostic indicator and a novel therapeutic target in HCC patients with HBV infection.

Background: Endoplasmic reticulum (ER) stress, with adaptive unfolded protein response (UPR), is a key link between obesity, insulin resistance and type 2 diabetes, all of which are often present in the most common endocrine-metabolic disorder in women of reproductive age, polycystic ovary syndrome (PCOS), which is characterized with hyperandrogenism. However, the link between excess androgen and Endoplasmic reticulum (ER) stress/insulin resistance in patients with polycystic ovary syndrome (PCOS) is unknown. Methods: An unexpected role of kisspeptin was reported in the regulation of UPR pathways and its involvement in the androgen-induced ER stress in hypothalamic neuronal cells. To evaluate the relationship of kisspeptin and ER stress, We detected Kisspeptin and other factors in blood plasm of PCOS patients, rat models and hypothalamic neuronal cells. Findings: We detected higher testosterone and lower kisspeptin levels in the plasma of PCOS than that in non-PCOS women. We established a PCOS rat model by dihydrotestosterone (DHT) chronic exposure, and observed significantly down-regulated kisspeptin expression and activated UPR pathways in PCOS rat hypothalamus compared to that in controls. Inhibition or knockdown of kisspeptin completely mimicked the enhancing effect of DHT on UPR pathways in a hypothalamic neuronal cell line, GT1-7. Kp10, the most potent peptide of kisspeptin, effectively reversed or suppressed the activated UPR pathways induced by DHT or thapsigargin, an ER stress activator, in GT1-7 cells, as well as in the hypothalamus in PCOS rats. Similarly, Kisspeptin attenuated thapsigargin-induced Ca2+ response and the DHT- induced insulin resistance in GT1-7 cells. Interpretation: Our data has revealed an unexpected protective role of kisspeptin against ER stress and insulin resistance in the hypothalamus and provided a new treatment strategy targeting hypothalamic ER stress and insulin resistance with kisspeptin as a potential therapeutic agent. Funding Statement: National Basic Research Program of China, National Natural Science Foundation of China (2012CB944902, 2012CB944703, 2013CB967404, 2013CB967401, 2013CB967403, 81571403, 81471429) and State Key laboratory of Reproductive Medicine (SKLRM-K201805). Declaration of Interests: The authors have no conflicts of interest about this work. Ethics Approval Statement: The study was approved by the Ethics Committee of Tongji University and all patients gave informed consent. All procedures in this study were approved by the Animal Experimentation Ethics Committee of the Chinese University of Hong Kong, and the methods were used in accordance with the guidelines of the Chinses University of Hong Kong.

Nicotinamide phosphoribosyltransferase (NAMPT) is a critical rate-limiting enzyme involved in NAD synthesis that has been shown to contribute to the progression of liver cancer. However, the potential role and mechanism of NAMPT in hepatitis B virus (HBV)-associated liver cancer remain unclear. The present study assessed the expression of NAMPT in HBV-positive and -negative liver cancer cells, and investigated whether HBV-induced NAMPT expression is dependent on HBV X protein (HBx). In addition, the role of NAMPT in HBV replication and transcription, and in HBV-mediated liver cancer cell growth was explored. The effects of NAMPT on the glycolytic pathway were also evaluated. Reverse transcription-quantitative PCR and western blotting results revealed that NAMPT expression levels were significantly higher in HBV-positive liver cancer cells than in HBV-negative liver cancer cells, and this effect was HBx-dependent. Moreover, the activation of NAMPT was demonstrated to be required for HBV replication and transcription. The NAMPT inhibitor FK866 repressed cell survival and promoted cell death in HBV-expressing liver cancer cells, and these effects were attenuated by nicotinamide mononucleotide. Furthermore, the inhibition of NAMPT was associated with decreased glucose uptake, decreased lactate production and decreased ATP levels in HBV-expressing liver cancer cells, indicating that NAMPT may promote the aerobic glycolysis. Collectively, these findings reveal a positive feedback loop in which HBV enhances NAMPT expression and the activation of NAMPT promotes HBV replication and HBV-mediated malignant cell growth in liver cancer. The present study highlights the important role of NAMPT in the regulation of aerobic glycolysis in HBV-mediated liver cancer, and suggests that NAMPT may be a promising treatment target for patients with HBV-associated liver cancer.

People with chronic obstructive pulmonary disease, cardiovascular disease, or hypertension have a high risk of developing severe coronavirus disease 2019 (COVID-19) and of COVID-19 mortality. However, the association between long-term exposure to air pollutants, which increases cardiopulmonary damage, and vulnerability to COVID-19 has not yet been fully established. We collected data of confirmed COVID-19 cases during the first wave of the epidemic in mainland China. We fitted a generalized linear model using city-level COVID-19 cases and severe cases as the outcome, and long-term average air pollutant levels as the exposure. Our analysis was adjusted using several variables, including a mobile phone dataset, covering human movement from Wuhan before the travel ban and movements within each city during the period of the emergency response. Other variables included smoking prevalence, climate data, socioeconomic data, education level, and number of hospital beds for 324 cities in China. After adjusting for human mobility and socioeconomic factors, we found an increase of 37.8% (95% confidence interval [CI]: 23.8%–52.0%), 32.3% (95% CI: 22.5%–42.4%), and 14.2% (7.9%–20.5%) in the number of COVID-19 cases for every 10-μg/m3 increase in long-term exposure to NO2, PM2.5, and PM10, respectively. However, when stratifying the data according to population size, the association became non-significant. The present results are derived from a large, newly compiled and geocoded repository of population and epidemiological data relevant to COVID-19. The findings suggested that air pollution may be related to population vulnerability to COVID-19 infection, although the extent to which this relationship is confounded by city population density needs further exploration., Graphical abstract Image 1, This research reported a national-level association with long-term exposure to air pollutants and COVID-19 cases covering 324 cities in China.

Introduction Vaccination coverage is widely used as metric of vaccination programme performance. However, VPDs outbreaks were reported in areas with high vaccination coverage. Timeliness and completeness have been considered more important assessment indicators of routine vaccination than overall vaccination coverage, but little is known in rural China. This study aimed to assess the timeliness and completeness of serial routine vaccinations among children in rural Southwest China. Methods A multi-stage stratified cluster survey was conducted among 1062 children aged 18–48 months in rural Guangxi. Vaccination status was obtained from child’s vaccination certificate. We calculated timely vaccination coverage, complete vaccination coverage, timely-and-complete vaccination coverage and 95% CI for routine vaccination through weighted estimation analysis. Weighted Kaplan-Meier analyses were applied to estimate the median delay periods for each dose of serial routine vaccines, including one-dose BCG, three-dose HepB, three-dose OPV, four-dose DTP, two-dose MCV, two-dose JEV and two-dose MPV-A. Complete coverage, and timely-and-complete coverage for combined 5-vaccine series were calculated. Results For each dose of routine vaccines, overall vaccination coverages were over 90%, but timely vaccination coverage ranged from the lowest of 44.4% for JEV1 to the highest of 92.5% for MPV-A1. For multi-dose routine vaccines, complete vaccination coverages varied from the lowest of 92.9% for MCV to the highest of 100% for HepB, and timely-and-complete vaccination coverages were lower than 80%, ranging from the lowest of 30% for JEV to the highest of 77.2% for MPV-A. For combined 5-vaccine series, complete coverage was 77%, while timely-and-complete coverage was 12.1%. MPV-A1 had the longest median delay of 176 days, but BCG and HepB1 had the shortest of 1 day. Conclusions The overall coverages of serial routine vaccinations were high, but the timeliness and completeness were poor. Relevant agencies of vaccination service should address timeliness-and-completeness into the assessment indicators of routine vaccination service quality.

The space-borne measured fine-mode aerosol optical depth (fAOD) is a gross index of column-integrated anthropogenic particulate pollutants, especially over the populated land. The fAOD is the product of the AOD and the fine-mode fraction (FMF). While there exist numerous global AOD products derived from many different satellite sensors, there have been much fewer, if any, global FMF products with a quality good enough to understand their spatiotemporal variations. This is key to understanding the global distribution and spatiotemporal variations of air pollutants, as well as their impacts on global environmental and climate changes. Modifying our newly developed retrieval algorithm to the latest global-scale Moderate Resolution Imaging Spectroradiometer (MODIS) aerosol product (Collection 6.1), a global 10-year FMF product is generated and analyzed here. We first validate the product through comparisons with the FMF derived from Aerosol Robotic Network (AERONET) measurements. Among our 169,313 samples, the satellite-derived FMFs agreed with the AERONET spectral deconvolution algorithm (SDA)-retrieved FMFs with a root-mean-square error (RMSE) of 0.22. Analyzed using this new product are the global patterns and interannual and seasonal variations of the FMF over land. In general, the FMF is large (>0.80) over Mexico, Myanmar, Laos, southern China, and Africa and less than 0.5 in the Sahelian and Sudanian zones of northern Africa. Seasonally, higher FMF values occur in summer and autumn. The linear trend in the satellite-derived and AERONET FMFs for different countries was explored. The upward trend in the FMFs was particularly strong over Australia since 2008. This study provides a new global view of changes in FMFs using a new satellite product that could help improve our understanding of air pollution around the world.

Background Identifying young individuals living with human immunodeficiency virus (HIV) who are unaware of their status is a major challenge for HIV control in China. To address this, an innovative, anonymous vending machine–based urine self-collection for HIV testing (USCT) program was implemented in 2016 in colleges across China. Methods From June 2016 to December 2019, 146 vending machines stocked with urine self-collection kits were distributed on 73 college campuses across 11 provinces of China. Urine samples were collected, delivered, and tested in an anonymous manner. We analyzed the returned rate, reactive rate (likelihood of HIV screening positive), testing effectiveness (the annual number of college students living with HIV screened by USCT or other testing methods), and the spatiotemporal relationship between USCT usage and student activity per college generated from the usage of a social networking application. Results Among the 5178 kits sold, 3109 (60%) samples were returned; of these, 2933 (94%) were eligible for testing. The HIV reactive rate was 2.3% (66 of 2933). The average effectiveness ratio among the 34 participating Beijing colleges was 0.39 (12:31) between USCT and conventional testing methods. A strong spatiotemporal correlation between USCT numbers and online student activity was observed during school semesters in Beijing. Conclusions USCT is a powerful complement to current interventions that target at-risk students and promote HIV testing. The social networking–based evaluation framework can be a guide in prioritizing at-risk target populations.

Objective To assess the efficacy and safety of retention enema with traditional Chinese medicine (TCM) for ulcerative colitis (UC) through a meta-analysis of published studies. Methods Literatures were retrieved from five electronic databases. Quality evaluation and meta-analysis were respectively conducted using the Cochrane collaboration and RevMan5.3. Overall quality of evidence was evaluated using GRADE system. Effect sizes were pooled using random effect models. Results Seventeen RCTs were included. Compared with routine pharmacotherapies (RPs), TCM enema exhibited a statistically significant difference in clinical efficacy and reduction of the recurrence rate. The results of qualitative description for other endpoints, such as improvements in anabrosis, ulcer, diarrhea, and hematochezia, suggested that TCM enema had better efficacy than RPs. Furthermore, the incidence of side effects in TCM was lower than that in RPs. Conclusion This meta-analysis confirmed the efficacy and safety of TCM enema for improving UC symptoms. However, further well-designed researches are needed.

Thyroid cancer is the most common endocrine carcinoma, with papillary thyroid carcinoma (PTC) accounting for 80%-90% of thyroid cancers. Accumulating studies reported that mitochondria plays an important role in the regulation of cell proliferation. ALDH5A1, may function as an oncogene or tumour suppressor in various human cancers, and the role of ALDH5A1 in PTC is still unclear. The aim of this study was to investigate the clinical significance of ALDH5A1 expression and its functions in PTC. In this present study, we studied ALDH5A1 expression on primary papillary thyroid carcinoma (PTC) in The Cancer Genome Atlas (TCGA) database. Results showed that the levels of ALDH5A1 were found positively correlated with tumour stage, metastasis, lymph node stage, and higher levels of ALDH5A1 demonstrated poor disease-free survival (DFS). Immunohistochemistry (IHC) revealed that significantly higher expression of ALDH5A1 was found in PTC tissues. On the other hand, knockdown of ALDH5A1 significantly inhibited PTC cell proliferation, migration and invasion detection found the migration and invasion of cells also were hindered when ALDH5A1 level was reduced. The knockdown of ALDH5A1 inhibited the expression of Vimentin and promoted the expression of E-cadherin. In brief, knockdown of ALDH5A1may act as a novel molecular target for the prevention and treatment of PTC. SIGNIFICANCE OF THE STUDY: The present study focused on the role and the potential mechanism of ALDH5A1 in papillary thyroid carcinoma. We demonstrated that reduced expression of ALDH5A1 might inhibit the progression of TC by inhibiting cell proliferation, migration and invasion and reversing epithelial-mesenchymal transition (EMT). The findings ensured the interaction relation between ALDH5A1 and EMT in PTC, providing a novel biological marker for PTC and enriching the potential strategies for TC treatment.

People with chronic obstructive pulmonary disease, cardiovascular disease or hypertension have a high risk of severe coronavirus disease 2019 (COVID-19). Long-term exposure to air pollution, especially PM2.5, has also been associated with COVID-19 mortality. We collated individual-level data of confirmed COVID-19 cases during the first wave of the epidemic in mainland China. We fitted a generalized linear model using city-level COVID-19 cases and severe cases as the outcome, and long-term average levels of air pollutants as the exposure. Our analysis was adjusted using several variables, including a mobile phone dataset, covering human movement from Wuhan before the travel ban and movements within each city during the time of emergency response. Other variables included census, smoking prevalence, climate, and socio-economic data from 324 cities in China. We adjusted for human mobility and socio-economic factors, and found that an increase in long-term NO2 or PM2.5 may correspond to an increase in the number of COVID-19 cases and severe infections. However, the linkage might also be affected by the confounding factor of population size because of the predefined correlation between population size and air pollution. The results are derived from a large, newly compiled and geocoded repository of population and epidemiological data relevant to COVID-19. The findings of this paper (and other previous studies that have given ambiguous results) indicate that a more definitive analysis is needed of the link between COVID-19 and air pollution.

As a GSK-3β inhibitor reported by our group, K313 is a novel benzoxazole derivative and displays anti-inflammatory properties in RAW264.7 macrophages without cytotoxicity. The activity of GSK-3β affects the differentiation and maturation of bone marrow-derived dendritic cells (DCs). This study aims to investigate whether K313 can be used to induce regulatory/tolerogenic dendritic cells (DCregs), and the therapeutic effects of DCregs induced by K313 in the autoimmune model of experimental autoimmune encephalitis (EAE). The results show that compared with LPS stimulated mature DCs, K313-treated bone marrow-derived DCs display obvious tolerogenic characteristics with decreased expression of co-stimulatory molecules, downregulated secretions of pro-inflammatory cytokines and unregulated secretion of anti-inflammatory cytokine IL-10. The above characteristics conform to the typical phenotypes of DCregs. Moreover, K313-modified DCregs inhibit antigen-specific T cell responses in vitro. Furthermore, by adoptive transfer, K313 modified DCregs to the EAE mice, and the development of disease was ameliorated to some extent. In addition, treatment with K313-modified DCregs also significantly reduced the percentages of splenetic Th1 and Th17 cells and increased the percentage of regulatory T cells in EAE mice. In conclusion, K313-modified DCregs show anti-inflammatory properties in vitro and have a significant positive effect on the EAE disease in vivo. Our data indicate that K313-induced DCregs pulsed with auto-antigen might have potential use as a therapeutic approach for autoimmune inflammation of the central nervous system.

The mechanism of cognitive dysfunction in diabetes is still unclear. Recently, studies have shown that the cerebellum is involved in cognition. Furthermore, diabetes-induced cerebellar alterations is related to vascular changes. Therefore, we aimed to explore the roles of vascular function in diabetes-induced cerebellar damage and motor learning deficits. Type 1 diabetes was induced by a single injection of streptozotocin in Sprague-Dawley rats. Motor learning was assessed by beam walk test and beam balance test. The pathological changes of the cerebellum were assessed by Hematoxylin and eosin staining and Nissl staining. Apoptosis was evaluated by anti-caspase-3 immunostaining. Protein expression was evaluated by western blotting and double immunofluorescence. Our results have shown that motor learning was impaired in diabetic rats, coupled with damaged Purkinje cells and decreased capillary density in the cerebellum. In addition, the protein expression of neuronal NOS, inducible NOS, endothelial NOS, total nitric oxide, vascular endothelial growth factor and its cognate receptor Flk-1 was decreased in the cerebellum. Gastrodin treatment ameliorated neuronal damage and restored protein expression of relevant factors. Arising from the above, it is suggested that vascular dysfunction and NO signaling deficits in the cerebellum may be the underlying mechanism of early manifestations of cognitive impairment in diabetes, which could be ameliorated by gastrodin intervention.

Infants with intrauterine growth retardation (IUGR) have a high risk of developing bronchial asthma in childhood, but the underlying mechanisms remain unclear. This study aimed to disclose the role of vascular non-inflammatory molecule 1 (vannin-1, encoded by the Vnn1 gene) and its downstream signaling in IUGR asthmatic mice induced by ovalbumin. Significant histological alterations and an increase of vannin-1 expression were revealed in IUGR asthmatic mice, accompanied by elevated methylation of Vnn1 promoter regions. In IUGR asthmatic mice, we also found (i) a direct binding of HNF4α and PGC1α to Vnn1 promoter by ChIP assay; (ii) a direct interaction of HNF4α with PGC1α; (iii) upregulation of phospho-PI3K p85/p55 and phospho-AktSer473 and downregulation of phospho-PTENTyr366, and (iv) an increase in nuclear NFκB p65 and a decrease in cytosolic IκB-α. In primary cultured bronchial epithelial cells derived from the IUGR asthmatic mice, knockdown of Vnn1 prevented upregulation of phospho-AktSer473 and an increase of reactive oxygen species (ROS) and TGF-β production. Taken together, we demonstrate that elevated vannin-1 activates the PI3K/Akt/NFκB signaling pathway, leading to ROS and inflammation reactions responsible for asthma occurrence in IUGR individuals. We also disclose that interaction of PGC1α and HNF4α promotes methylation of Vnn1 promoter regions and then upregulates vannin-1 expression., Summary: Vascular non-inflammatory molecule 1 (vannin-1) is elevated in intrauterine growth retardation (IUGR) asthmatic mice, leading to activation of the PI3K/Akt/NFκB pathway responsible for bronchial oxidative stress and hyperresponsiveness.

Context: Ginsenoside Rg1 (Rg1), as the active ingredient of Panax notoginseng, has protective effect on the cardiovascular system. While, the efficiency of Rg1 against acute myocardial infarction (MI) is not fully elucidated. The aim of this study was to investigate the cardio-protection and the underlying mechanism for Rg1. Objective: Myocardial infarction was induced by ligation of the left anterior descending coronary artery of wistar rats. 2,3,5-triphenyltetrazolium chloride (TTC) stain assay was used to detect infarct size; hematoxylin and eosin (HE) and phosphotungstic acid-hematoxilin (PTAH) stain assay were used to evaluate the cardiac structure; terminal deoxytransferase-mediated dUTP-biotin nick end labeling (TUNEL) assay was used to detect apoptosis; immunohistochemical stain was used to evaluate neutrophils and macrophages infiltration. Key Findings: Firstly, Extract Rg1 (P < 0.01) and Ferment Rg1 (P < 0.01) down-regulated myocardial infarct size and up-regulated superoxide dismutase (SOD) activity significantly. Then, HE and PTAH stain indicated the protection of Extract Rg1 and Ferment Rg1 on heart structure, especially on the integrity of sarcomere. Finally, both Extract Rg1 (P < 0.001) and Ferment Rg1 (P < 0.001) reduced the number of apoptosis cells, and further inhibited the infiltration of neutrophils into the infarct site of heart. Summary: For the first time, our present study verified the efficiency of Rg1 against acute MI, and provided experimental evidence that Ferment Rg1 holds similar cardio-protection with Extract Rg1. Rg1 holds potential therapeutic value for further drug development on heart disease.

Nonalcoholic fatty liver (NAFLD) is associated with metabolic abnormalities and changes in lipid processing. Fatty acids (FAs) are important signaling molecules in the body and are the main raw materials for lipid synthesis. To explore the lipidomic profile of serum fatty acids in mice, we put the mice on a high-fat diet (HFD) to induce NAFLD. We applied a targeted metabolic analysis approach using gas chromatography mass spectrometry (GC-MS) and established a method for simultaneous determination of 36 medium and long-chain fatty acids in the serum of mice. This method was validated for linearity, range, limit of detection (LOQ), limit of quantification (LOD), precision, accuracy, and recovery. The levels of most FA species and total FAs were increased in the HFD group (n=9) compared with the lean group (n=9). The principal component analysis (PCA) and the orthogonal partial least squares discriminant analysis (OPLS-DA) of all FA species were analyzed in both groups. We found that palmitic acid (16:0), stearic acid (18:0), oleic acid (18:1), and arachidonic acid (20:4) were significantly different (VIP＞1, p＜0.05 and FC＞1.5) in the two groups. We also calculated serum indices measure the activities of different enzymes involved in fatty acid metabolism. The de novo lipogenesis index, elongase index, n-6 FAs, saturated FAs, unsaturated FAs, and SCD1 index 2 were increased in the HFD group compared with the lean group while the n-3 FAs, n-3/n-6 of the index, and SCD1 index 1 were decreased in the HFD group compared with the lean group. Relationships between major FA species and biochemical indicators (LDL-C, TC, TG, ALT, and AST) were evaluated by Pearson analysis. The serum FA concentrations (C16:0, C18:0, C18:1, and C20:4) and serum indices ( De novo lipogenesis index, elongase index, saturated FAs, unsaturated FAs and n-6 FAs) positively correlated with LDL-C, TC, TG, ALT, and AST. Collectively, our study provides insights into NAFLD and its effect on lipid metabolism.

Particulate matter 2.5 (PM_(2.5)) pollution has long been a global environmental problem and still poses a great threat to public health. This study investigates global spatiotemporal variations in PM_(2.5) using the newly developed satellite-derived PM_(2.5) dataset from 1998 to 2018. An integrated exposure–response (IER) model was employed to examine the characteristics of PM_(2.5)-related deaths caused by chronic obstructive pulmonary disease (COPD), ischemic heart disease (IHD), lung cancer (LC), and stroke in adults (age≥25), as well as lower respiratory infection (LRI) in children (age≤5). The results showed that high annual PM_(2.5) concentrations were observed mainly in East Asia and South Asia. Over the 19-year period, PM_(2.5) concentrations constantly decreased in developed regions, but increased in most developing regions. Approximately 84% of the population lived in regions where PM_(2.5) concentrations exceeded 10 μg/m³. Meanwhile, the vast majority of the population (>60%) in East and South Asia was consistently exposed to PM_(2.5) levels above 35 μg/m³. PM_(2.5) exposure was linked to 3.38 (95% UI: 3.05–3.70) million premature deaths globally in 2000, a number that increased to 4.11 (95% UI: 3.55–4.69) million in 2018. Premature deaths related to PM_(2.5) accounted for 6.54%−7.79% of the total cause of deaths worldwide, with a peak in 2011. Furthermore, developing regions contributed to the majority (85.95%–95.06%) of PM_(2.5)-related deaths worldwide, and the three highest-ranking regions were East Asia, South Asia, and Southeast Asia. Globally, IHD and stroke were the two main contributors to total PM_(2.5)-related deaths, followed by COPD, LC, and LRI.

Phosphonates, featuring organic skeletons binding phosphonic acid group, are ubiquitous in wastewaters and natural waters. Highly efficient removal of trace phosphonates is desirable to ensure the water quality and to prevent eutrophication. Commercially available hyper-cross-linked resins modified by amino groups have been proved to exhibit preferable adsorption of various organic acids from aqueous systems via synergetic effect of various interactions (electrostatic interaction, van der Waals interaction, π-π interaction, etc.) but seem relatively ineffective for phosphonates. In this study, we proposed a new strategy via coupling a commercial resin NDA88 with nanoconfinement of Hydrated ferric oxide (HFO) for selective removal of phosphonates. The resultant HFO@NDA88 shows an adsorption capacity of 15.0 mg P/g towards a model phosphonate (nitrilotris(methylenephosphonic acid)), higher than those of the NDA88 host (7.9 mg P/g) and the bare HFO (7.4 mg P/g). Remarkably, compared to NDA88 and HFO, HFO@NDA88 exhibits much higher resistance to the interference of various anions and other substances at much greater levels, particularly the organic analogue nitrilotriacetic acid and phosphate. Such superior selectivity is believed to result from the cooperative contributions of the host NDA88 as well as the embedded nano-HFO. Moreover, HFO@NDA88 shows excellent reusability and satisfactory performance in fixed-bed water treatment. This work provides a rational design of nanocomposite adsorbents for highly selective removal of phosphonates from complicated water matrix.

Objective To investigate the prognostic significance of and risk factors for solitary lymph node metastasis (SLNM) of patients with cervical carcinoma. Methods Clinical data from patients with International Federation of Gynecology and Obstetrics (FIGO) stages IA2 to IIA cervical carcinoma who underwent radical hysterectomy and pelvic lymphadenectomy between January 2003 and December 2010 were analysed retrospectively. Histopathological analysis was used to identify SLNM. Long-term survival and risk factors associated with SLNM were analysed. Results The study enrolled 302 patients with cervical cancer: 48 with SLNM (SLNM group) and 254 patients with no lymph node metastases (nLNM group). FIGO stage, tumour grade, depth of tumour invasion, uterine body involvement, parametrial involvement and lymphovascular invasion differed significantly between the two groups. Logistic regression analysis revealed that FIGO stage, depth of tumour invasion and lymphovascular invasion were independent factors associated with SLNM. The 5-year survival rates of the SLNM and nLNM groups were 54.2% and 87.8%, respectively. Multivariate analysis identified SLNM as an independent factor affecting survival. Conclusions The occurrence of just one solitary lymph node metastasis significantly worsened the prognosis in patients with cervical carcinoma compared with patients without lymph node metastases.

Human intestinal bacteria play an important role in the metabolism of herbal medicines, leading to the variations in their pharmacological profile. The present study aimed to investigate the metabolism of Xiao-Cheng-Qi decoction (XCQD) by human intestinal bacteria and to discover active component combination (ACC) contributing to the anti-inflammatory activity of XCQD. The water extract of XCQD was anaerobically incubated with human intestinal bacteria suspensions for 48 h at 37 °C. A liquid chromatography-hybrid quadrupole time-of-flight mass spectrometry (LC-Q-TOF/MS) method was performed for identification of the metabolites. In addition, the anti-inflammatory effects of XCQD and biotransformed XCQD (XCQD-BT) were evaluated in vitro with cytokines in RAW264.7 cells induced by lipopolysaccharide (LPS). A total of 51 compounds were identified in XCQD and XCQD-BT. Among them, 20 metabolites were proven to be transformed by human intestinal bacteria. Significantly, a combination of 14 compounds was identified as ACC from XCQD-BT, which was as effective as XCQD in cell models of inflammation. In conclusion, this study provided an applicable method, based on intestinal bacterial metabolism, for identifying combinatory compounds responsible for a certain pharmacological activity of herbal medicines.

Termite gut microbiome is a rich reservoir for glycoside hydrolases, a suite of enzymes critical for the degradation of lignocellulosic biomass. To search for hemicellulases, we screened 12,000 clones from a fosmid gut library of a higher termite, Globitermes brachycerastes. As a common Southeastern Asian genus, Globitermes distributes predominantly in tropical rain forests and relies on the lignocellulases from themselves and bacterial symbionts to digest wood. In total, 22 positive clones with β-xylosidase activity were isolated, in which 11 representing different restriction fragment length polymorphism (RFLP) patterns were pooled and subjected to 454 pyrosequencing. As a result, eight putative β-xylosidases were cloned and heterologously expressed in Escherichia coli BL21 competent cells. After purification using Ni-NTA affinity chromatography, recombinant G. brachycerastes symbiotic β-xylosidases were characterized enzymatically, including their pH and temperature optimum. In addition to β-xylosidase activity, four of them also exhibited either β-glucosidase or α-arabinosidases activities, suggesting the existence of bifunctional hemicellulases in the gut microbiome of G. brachycerastes. In comparison to multimeric protein engineering, the involvement of naturally occurring multifunctional biocatalysts streamlines the genetic modification procedures and simplifies the overall production processes. Alternatively, these multimeric enzymes could serve as the substitutes for β-glucosidase, β-xylosidase and α-arabinosidase to facilitate a wide range of industrial applications, including food processing, animal feed, environment and waste management, and biomass conversion.

To evaluate gait characteristics and investigate changes pre- and post- cerebrospinal fluid tap test (CSF TT) in gait parameters in patients with probable idiopathic normal pressure hydrocephalus (iNPH).Sixty patients were sequential circuited and diagnosed with possible iNPH according to Japanese second iNPH guidelines at our hospital from December 2016 to March 2021. All patients underwent the CSF TT. Gait parameters, cognitive and urinary function were assessed pre- and post-CSF TT. Patients who were unable to ambulate to take the tests or could not walk independently or walked normally were excluded.Twenty-six patients were diagnosed with probable iNPH using the CSF TT. After CSF TT, the Boon sum score improved from 20.0 ± 7.7-16.6 ± 8.0 (p 0.001), the Boon walking score improved from 8.9 ± 3.5-7.8 ± 4.4 (p = 0.008), the Boon step score improved from 6.3 ± 2.3-5.2 ± 2.1 (p 0.001), the Boon time score improved from 4.9 ± 2.4-3.7 ± 2.3 (p 0.001), tandem walking disturbance improved from 1.7 ± 0.7-1.4 ± 0.9 (p = 0.043), tendency toward falling improved from 1.7 ± 0.7-1.3 ± 1(p = 0.022), 3-meter timed up and go test (3-mTUG) improved from 21.9 ± 7.1-17.6 ± 5.1( p 0.001), 10-meter walking (10-MWT) step improved from 31.1 ± 13.1-24.6 ± 7.5 (p 0.001), velocity improved from 0.7 ± 0.2-0.8 ± 0.3 (p 0.001) and stride length improved from 0.4 ± 0.1-0.46 ± 0.1(p 0.001), compared with before the CSF TT.These results suggest that many parameters in the Boon gait test were responsive to the CSF TT, and the Boon gait test may help objectify response to the CSF TT by combining the 10-MWT and 3-mTUG gait assessments.