Previous research pointed to a close relationship between the incidence of tuberculosis (TB) in aging populations and socio-economic conditions, however there has been lack of studies focused on a region of unbalanced socio-economic development. The aim of this paper is to explore the spatio-temporal variation in TB incidence and examine risk determinants of the disease among aging populations in a typical region.q statistic) of risk factors for TB among the elderly.Data on TB-registered cases between 2009 and 2014, in addition to social-economic factors, were collected for each district/county in Beijing, Tianjin and Hebei, a region characterized by an aging population and disparities in social-economic development. A Bayesian space–time hierarchy model (BSTHM) was used to reveal spatio-temporal variation in the incidence of TB among the elderly in this region between 2009 to 2014. GeoDetector was applied to measure the determinant power (q = 0.20) and medical conditions (q = 0.17). These factors had a non-linear interactive effect on the incidence of TB among the elderly, with medical conditions and the level of economic development having the strongest effect (q = 0.54).The incidence of TB among the elderly exhibited geographical spatial heterogeneity, with a higher incidence in underdeveloped rural areas compared with that in urban areas. Hotspots of TB incidence risk among the elderly were mostly located in north-eastern and southern areas in the study region, far from metropolitan areas. Areas with low risk were distributed mainly in the Beijing-Tianjin metropolitan areas. Social-economic factors had a non-linear influence on elderly TB incidence, with the dominant factors among rural populations being income (The findings explain spatio-temporal variation in TB incidence and risk determinants of elderly TB in the presence of disparities in social-economic development. High-risk zones were located mainly in rural areas, far from metropolitan centres. Medical conditions and the economic development level were significantly associated with elderly TB incidence, and these factors had a non-linear interactive effect on elderly TB incidence. The findings can help to optimize the allocation of health resources and to control TB transmission in the aging population in this region. Mycobacterium tuberculosis complex; only pulmonary TB is contagious [Tuberculosis (TB) is an airborne infectious disease caused by the ntagious . It is antagious . Accordintagious . The eldntagious .TB has been considered a global public health emergency for the past decades, and much progress in reducing the incidence of TB has been made, however, no country has yet eliminated TB . AlthougThe demographic structure of a population is a key determinant of infectious disease spread . In sociAccording to some research, regional TB incidence is closely associated with poverty, poor living conditions and human agglomeration . PovertyUsing various spatial statistical methods, previous research confirmed that the risk of TB transmission is spatially heterogeneous. These methods included Moran’s I and spatial panel data models , Kernel There were two primary aims of the present study: The first aim was to investigate spatio-temporal variation in the incidence of TB among individuals older than 65 years in the Beijing-Tianjin-Hebei urban agglomeration, which is a region with a high proportion of aging households and socio-economic disparities. The second aim was to investigate whether there was a non-linear association between the risk of TB in this aging population and socio-economic factors.The study region was the North China Plain and consisted of the Beijing Municipality, Tianjin Municipality and Hebei Province. This region shows spatial heterogeneity in economic development. Beijing is the capital of China, Tianjin is one of a number of municipalities directly under the control of central government. Two neighbouring cities constitute a large metropolis. Surrounding the two cities is Hebei Province.Economic development in the region is unbalanced, with many of the poorest counties in the country during the study period located in Hebei Province. In addition, aging of the population is becoming a growing concern in the study region, as the proportion of the population aged older than 65 years in Beijing, Tianjin and Hebei is 8.72, 8.53 and 8.69%, respectively. This index is an important international criterion used to measure the degree of population aging in a country or a region, with values over 7% constituting an aging society.In this study, data on TB cases in Beijing, Tianjin and Hebei were obtained from the Chinese Center for Disease Control and Prevention, which is a national real-time surveillance system, and the Notifiable Disease Reporting System, which covers all districts and counties in the study region. In total, data on 48,213 TB patients older than 65 years were collected. The study period was 2009–2014. Figure Data on potential factors related to TB in the region in 2009–2014 were collected from the statistical yearbook, which includes information on Gross Domestic Product (GDP) per capita, population density, industrial structure, urban and rural household income and number of beds in medical facilities  and the relative risk (RR) of elderly TB incidence (rit):In this study, new TB cases α is a constant term and si is a spatial term expressing the spatial distribution of elderly TB risk factors across each county in the study region throughout the whole study period. This spatial term represents spatial heterogeneity in elderly TB incidence risk, and its posterior estimated exp (si) describes the spatial risk of elderly TB in county i relative to that in the whole region. b0t* + vt is a temporal term, expressing the temporal distribution of elderly TB risk factors across each year of the study period in the whole region. Its posterior estimation exp (b0t* + vt) describes the overall temporal RR in the study region, t* describes the the middle of the years range, and vt represents Gaussian noise, indicating a random time effect. The parameter bi is the local trend in county i and represents the spatial heterogeneity of a temporal trend, expressing the spatio-temporal interaction effect. It can be used to measure deviation from common spatio-temporal variation. If the value of bi is greater than 1, this It indicates that county i shows a stronger temporal trend compared with the overall trend of the whole region and vice versa. The term ɛ1i represents variation not yet explained by the model. In the study, it is assumed to have a Gaussian distribution.A log link regression function was applied to assess spatio-temporal variation in elderly TB incidence in the BSTHM model . The funp (exp (si) > 1 | data). If the probability of RR in a county was greater than 0.8, it was defined as a hotspot; if the probability of RR in a county was less than 0.2, it was defined as a cold spot; if the probability of RR in a county was between 0.2 and 0.8, the county was defined as neither a hotspot nor a cold spot. Local temporal trends in each spot-class region were then further analysed. In each county, the temporal trend b1i was stratified into three classes  compared with the overall trend (b0t* + vt) based on the posterior probability p. Regions where temporal trends were decreasing faster had a probability greater than 0.8, regions where temporal trends were decreasing slower had a probability less than 0.2, and regions where temporal trends were stable had a probability of between 0.2 and 0.8. The BSTHM was implemented in WinBUGS software [Based on the posterior estimated parameters of the BSTHM, a two-stage method was used to identify spatial heterogeneity in elderly TB RR and to analyse local temporal trends. Spatial RR was first stratified into three classes: hotspot, cold spot or not hot/cold spot, based on the posterior probability software .q statistic of GeoDetector have facilitated investigations of spatial non-linear associations in spatial-temporal data. GeoDetector is a spatial variance analysis method, which can be used to assess the power of an effect factor and analyse the nature of the interactive between a pair of factors. The core concept of the method is that if a potential factor is related to a disease, this factor will be linked to the spatio-temporal distribution of the disease [Recent advances in the  disease –26. Thus disease , 28.q statistic of GeoDetector was used to assess spatio-temporal associations between elderly TB and potential risk factors. The GeoDetector q statistic was expressed as:N and Nh are the number of counties in the study region and in the h-th stratum , respectively. σ2 represents the variance in TB incidence in all the counties in the study region, and σh2 is the stratified variance in TB incidence in all the counties in the h-th stratum. The parameter Ri is the TB incidence in i-th county, and Rh,j is the TB incidence in j-th county in the h-th stratum. The q statistic represents the non-linear association between a potential factor and elderly TB incidence. The value lies between 0 and 1, with higher values of the q statistic indicating higher determinant power of the variable. If q = 0, there is no association between elderly TB incidence and the variable. In contrast, if q = 1, then elderly TB incidence is completely determined by the variable. In the study, all the factors in Table The 5, with apparent spatio-temporal heterogeneity. Spatially, the highest incidence (i.e. above 300/105), was found mainly in the northwest and southeast of the study region, whereas the lowest incidence (i.e. below 1/105) was found mainly in the Beijing and Tianjin megalopolis and its surrounding regions. Temporally, the elderly TB incidence in the region decreased gradually, with an incidence of 99.6/105 in 2009 versus 80.4/105 in 2014.From 2009 to 2014 in the study region, the average annual TB incidence was 87.6/10Spatial risk factors for elderly TB incidence were calculated using the BHSTM. The posterior medians of the spatial RRs in the counties, shown in Fig. The common spatial RR of elderly TB incidence was not stable. The results indicated that the RR of elderly TB incidence in all the counties in the study region showed an overall decreasing trend from 2009 to 2014 Fig. . In 2009Figures The local temporal trend of hotspots is shown in Fig. The local temporal trend in cold spots is shown in Fig. q statistic calculation. Seven potential socio-economic risk factors were selected as explanatory variables: per capita GDP, population density, proportion of primary industry in GDP, proportion of tertiary industry in GDP, per capita disposable income of urban residents, per capita disposable income of rural households and beds in health care institutions per 1000 persons. Using these variables, the risk factors and their interaction effects on elderly TB incidence were assessed . A large number of beds in health care institutions was associated with a low incidence of elderly TB cases, with a q value of 0.17 (p < 0.01). Population density, income of urban residents and per capita GDP had relatively low effects on elderly TB incidence, with q values of 0.16, 0.16 and 0.14, respectively.The results showed that socio-economic and health condition factors were the dominant determinants of spatio-temporal variation in elderly TB incidence. Income of rural households showed the strongest association with elderly TB incidence. Low income of rural households was associated with a high incidence of elderly TB incidence, with a q statistic for the interaction was 0.54 (p < 0.01). There was also an interaction between health conditions and population density , health conditions and population density  and health conditions and rural and urban income .Analysis of interaction effects indicated that health conditions and the proportion of primary industry in GDP had a combined effect on elderly TB risk. The We designed the present study to analyse spatio-temporal variation in TB risk in the over-65 population and to detect potential factors influencing TB risk in a region of unbalanced socio-economic development in China (i.e. Beijing-Tianjin-Hebei). The results revealed the highest risk mainly in rural areas in the northeast and southwest, far from metropolitan areas. Interaction effects between health conditions and socio-economic factors had a significant effect on elderly TB incidence.In the study, elderly TB risk showed significant spatial heterogeneity, with a relatively high risk primarily in the northwest mountainous regions and south rural regions and a relatively low risk mainly in larger cities and surrounding areas. In terms of risk factors, the proportion of primary industry and tertiary industry in GDP was significantly associated with elderly TB incidence. This finding confirms the impact factors in regions with unbalanced socio-economic development.Our findings support those of other studies, which reported different urban and rural epidemiological characteristics of elderly TB patients, however, there were inconsistencies between these studies in terms of the TB risk level. For example, in a study carried out in Bangladesh, the authors reported higher TB-related mortality rates in urban than rural regions . FurtherIn this study, the regional economic development level was an important index of factors influencing TB transmission. Previous studies revealed an association between TB and per capita GDP, poverty, living standards and under-nutrition , 32. AccPer capita GDP can provide insights into the effect of in macro-level economic determinants and the effects of policy on spatio-temporal variations in elderly TB incidence. Previous studies indicated that per capita GDP was significantly associated with TB incidence at the country or provincial level , 33–35. According to many previous studies, income level is an important determinant of TB transmission, with TB closely associated with poverty and an increase in the proportion of TB cases in accordance with low income . Low incIn this study, population density was significantly associated with elderly TB incidence. Some previous studies reported a high TB incidence in areas of high population density , 37. ForThis study has some limitations. TB is a multifactorial disease. Thus, factors other than those considered in the present study , may have affected the incidence of TB in the study region. We used aggregate data on a county-level spatial scale in modelling. This may have introduced an ecological fallacy effect for some factors.Furthermore, new factors, such as epidemic of the other emerging infectious disease, will also influence the transmission of TB. Future research should consider additional factors that account for multi-level relationships in the incidence of TB.Our findings shed light on spatio-temporal variation in TB incidence and risk determinants of TB infection among the elderly in a region with disparities in socio-economic development. High-risk hotspots were located mainly in rural areas, far from metropolitan sectors, and medical conditions and economic development level were significantly associated with elderly TB incidence. These variables showed non-linear interactions with each other in influencing TB incidence. TB is of long standing, and the End TB Strategy, adopted by the WHO in 2014, includes targets for reductions in the TB disease burden in 2016–2035 [


Abdominal aortic calcification is a potentially important independent risk factor for cardiovascular health. The aim of this study was to determine the relationship between serum chloride level and abdominal artery calcification.We obtained the data of 3,018 individuals from the National Health and Nutrition Examination Survey database and analyzed the relationship between serum chloride and abdominal artery calcification. We performed stratified and single factor analysis, multiple equation regression analysis, smooth curve fitting, and threshold effect and saturation effect analysis. R and EmpowerStats were used for data analysis.Serum chloride is independently related to the AAC total 24 score (AAC-24). The smooth curves fitted were all inverted-U shaped. Below a cutoff value of 92 mmol/L, increase in serum chloride level was associated with increase in AAC-24; however, above that cutoff, increase in serum chloride level was associated with decrease in AAC-24.At serum levels below 92 mmol/L, chloride is a risk factor for abdominal aortic calcification but levels above 92 mmol/L appear to protect against abdominal aortic calcification. Aortic calcification is a risk marker for cardiovascular diseases such as coronary artery disease and stroke –4. FactoSerum chloride level has complex effects in humans. It is associated with several diseases, including hypertension , heart fThe aim of this study was to evaluate the relationship between serum chloride and abdominal aortic calcification using a representative sample of elderly individuals from the National Health and Nutrition Examination Survey (NHANES) .The study population was selected from the database of the NHANES—a population-based cross-sectional survey designed to collect information about the health and nutrition of the population of the US. The project surveys a nationally representative sample each year. These populations are located in counties across the country. NHANES interviews collect demographic, socioeconomic, dietary, and health-related information. Physical examinations conducted during the survey include physiological measurements and laboratory examinations .In this study, we retrospectively analyzed the data recorded for the period 2013–2014, which represents one cycle of the NHANES. From among the 10,175 individuals registered in the NHANES database during this period, we eliminated 7,035 because of missing data on AAC Total 24 score (AAC-24) and another 122 because of missing data on serum chloride level. The data of the remaining 3,018 individuals were included in this analysis. This study was approved by the Ethics Review Board of the National Center for Health Statistics. Informed consent was not considered necessary as this was an analysis of a database.The exposure variable for this study was serum chloride level (mmol/L). Blood specimens were processed and stored in the mobile examination center laboratory, and shipped to the Johns Hopkins University Lipoprotein Analytical Laboratory for analysis. The outcome variable was AAC-24, which was used to describe the amount of calcification. Several studies have shown that lateral spine images obtained with dual-energy X-ray absorptiometry (DXA) for vertebral fracture assessment can detect AAC with reasonably good sensitivity and specificity. The image resolution of lateral spine scans obtained with DXA is close that of a standard radiograph, and radiation exposure is much lower , 20. Forhttp://www.cdc.gov/nchs/nhanes/.The categorical variables included as covariates in our analysis were sex, race/ethnicity, and education level. The continuous covariates included in our analysis were age, body mass index (BMI), systolic blood pressure, diastolic blood pressure, total serum calcium (mmol/L), serum cholesterol (mmol/L), serum albumin (g/L), serum glucose, and refrigerated serum glucose (mmol/L). All data on serum chloride, AAC-24 score, and the covariates are available at http://www.r-project.org) and EmpowerStats . P < 0.05 was considered statistically significant.We performed weighted and variance estimation analyses to account for significant variance in our data set. Weighted multiple logistic regression models were used to assess the association between serum chloride and AAC-24. Weighted chi-square test was used to analyze differences in categorical variables between groups, and a weighted linear regression model was used to analyze differences in continuous variables. Stratified multiple regression was used for subgroup analysis. In addition, smooth curve fitting and generalized additive models were used to address the non-linear relationship between serum chloride and AAC-24. A recursive algorithm was used to calculate the inflection point in the relationship between serum chloride and AAC-24 when non-linearity was detected, with a bi-segmented linear regression model on either side of the inflection point. Data analysis was performed using R 3.6.3 /receptor activator of NF-κ B ligand system. The Wnt/β-Catenin pathway is activated or inhibited by lithium chloride  in vivo , implyin in vivo , 24. How in vivo , 24. In Our large sample size allowed subgroup analysis. When refrigerated serum glucose was 5.38–32.03 mmol/L, the smooth fitting curve between serum chloride and AAC-24 was inverted-U shaped, which is similar to the overall fitting curve between serum chloride and AAC-24, but the inflection point was >92 mmol/L. In the low serum glucose group (2.72–5.33 mmol/L), there was a downward fitting curve. In the low age triad (40–50 years), the smooth fitting curve between serum chloride and AAC-24 score was flat. The smooth fitting curves of low systolic blood pressure (74–116 mmHg) and high systolic blood pressure (76–122 mmHg) were flat.Since we used a nationally representative sample, our findings are generalizable to the whole US population. Our results suggest that serum chloride levels in adults should be maintained at an appropriate level, and that control of serum chloride level could be a new method for the prevention and control of peripheral artery calcification. Appropriate and individualized measures for prevention for AAC may have to be developed for different ethnic populations, sexes, and age-groups.It is important to recognize the limitations of our research. First, because of the cross-sectional design of our study, a causal relationship between adult serum chloride level and AAC-24 cannot be inferred. Second, confounding factors not analyzed in this study may affect the results. Third, no attempt has been made to clarify the mechanism by which serum chloride affects abdominal aortic calcification. Fourth, it would be more convincing if the range of the lowest and highest values of serum chloride among the participants examined were larger.Below a cutoff value of 92 mmol/L, serum chloride appears to be a risk factor for development of abdominal aortic calcification, but serum levels >92 mmol/L may protect against abdominal aortic calcification. Thus, detection of serum chlorine detection could be a simple and accurate method for screening individuals for risk of abdominal aorta calcification and cardiovascular disease.The datasets presented in this study can be found in online repositories. The names of the repository/repositories and accession number(s) can be found in the article/This study was approved by the Ethics Review Board of the National Center for Health Statistics. Informed consent was not considered necessary as this was an analysis of a database.SH, JP, and YW contributed to data collection, analysis, and writing of the manuscript. SH, TL, WZ, JY, DZ, YZ, SW, QG, LS, DY, JP, YW, and JX contributed to study design and writing of the manuscript. All authors read and approved the final manuscript.This study was supported by grants from the National Natural Science Foundation of China (81860379 and 82160410) and the Science and Technology Planning Project at the Department of Science and Technology of Jiangxi Province, China (20171BAB 205075).The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.


The data obtained revealed substantial variations in elemental composition between the test groups in comparison to the controls. However, the significant variations concerned only a few elements. The phosphorous (P) level and the ratio of P/Ca were significant in retinopathy and diabetes, whereas cobalt  was significant in AMD. In co-existing hypertension, the levels of tin , titanium , and ruthenium  varied in comparison to the controls. The study revealed inter-elemental interactions. The correlation matrices demonstrated the domination of the positive correlations, whereas negative correlations mainly concerned sodium.The aim of the study was the multi-elemental analysis of aqueous humor (AH) collected from patients undergoing cataract surgery. The study included: 16 patients with age-related macular degeneration AMD (99 controls), 10 patients with retinopathy (105 controls), 61 patients with hypertension (54 controls), and 33 patients with coexisting diabetes (82 controls). The control groups were recruited from patients with a lack of co-existing disease characterizing the specified studied group. The measurements were performed by the use of inductively coupled plasma optical emission spectrometry (ICP-OES). The statistical analysis was carried out using non-parametric testing (Mann–Whitney U). The level of significance was set at  Many adults experience significant vision loss because of their elderly age or common ocular diseases such as age-related macular degeneration (AMD), cataracts, diabetic retinopathy (DR), or glaucoma.Despite available treatments, their etiology still remains poorly understood. Therefore, there is a need to search for factors influencing eye health as well as uncover efficient prevention strategies.3−, and Cl−) in comparison to blood plasma appears to be much larger than might be expected from the Gibbs–Donnan equilibrium. Moreover, AH exhibits differences in protein composition in comparison with plasma.A cataract is a progressive loss of transparency in the eye’s lens, accounting for almost 50% of blindness in the world . So far,−1. AH was also tested for, among others, oxidative–antioxidant balance. The relationship between cataract maturity and markers of lipid peroxidation and activity of enzymatic antioxidants such as superoxide dismutase (SOD) and catalase (CAT) was assessed. Multiple logistic regression analyses identified them as predictors of cataract maturity [p = 0.006) with diabetic cataracts [Chowdhury et al.  identifimaturity . It was maturity ,10. The maturity . It was ataracts . Tryptopataracts . The obtataracts  proved t+/H+, Na+/Ca2+, HCO3−/Cl−) or active transport thanks to Na-K-ATPase [Besides components such as immunoglobulins, neuropeptides, proteins, lipids, carbohydrates, enzymes, the vitamins, AH was also examined for its elemental composition. It is known that electrolyte balance is of vital importance to lens transparency. Inorganic ions move through ion channels such as potassium, sodium, chloride, and calcium channels relying on ion exchange . In 2019, Kayiklik et al. [In accordance with the literature data, cataracts occur more frequently and at an earlier age in diabetic patients rather than in non-diabetic ones ,25. The k et al.  examinedStopa et al.  examinedHohberger Bettina et al.  establisThe levels of macro-and micronutrients change dynamically in the human body, and it is indisputable that these are of critical importance to human health. Many studies concern the determination of the level of both essential and toxic metals in different tissues, taken from patients suffering from various diseases, including eye problems. However, the results of these studies appear problematic, chiefly due to the variety of methods used . The inconsistency of the results obtained by different teams be due to the criteria of selecting the test and control groups or the unquestionable inter-patient variability.Taking into account the individual variability of patients and the inability to collect a control group composed only of healthy volunteers, the research findings still need to be verified on new and larger groups of patients inhabiting different environments and suffering from varied coexisting diseases. The aim of the current study was to evaluate the elemental composition of AH in patients undergoing cataract surgery, additionally suffering from diabetes, hypertension, or diabetic retinopathy. The control group consisted of patients undergoing cataract surgery without distinguished co-morbidities. Statistical analysis of the results will allow for the hypothesis to be verified regarding the existence of differences (or their lack) in the composition of the eye fluid of cataract patients between the selected subgroups.p = 0.05. In the present study, AH samples were examined by using ICP-OES with the aim to measure the concentration of 67 elements that were divided into 5 clusters according to the previous report . All meaMany toxic metals have been found in examined samples such as mercury (Hg), thallium (Tl), cadmium (Cd), aluminum (Al), or lead (Pb). Pb was found in all samples studied at the range from 0.56 to 1.66 ppm, Al was in the range from 1.03 to 2.59 ppm, and Cd occurred at a much lower concentration, from 0 to 0.013 ppm. AH toxic metals’ concentrations reflect an exposure of all patients to these metals, which are known to be permanent industrial pollutants standing behind the environmental contamination. Despite the fact that there are many pieces of evidence indicating that these metals can cause oxidative stress by ROS exert production, resulting, among others, in lipid peroxidation, DNA damage, and the depletion of cellular antioxidant defense systems ,30,31, nThe highest concentration range was observed for Ca, Cs, K, Mg, Na, P, and Rb. p > 0.05). Only the concentration of phosphorous (P) in the AH of patients suffering from diabetes (p = 0.017) and diabetic retinopathy (p = 0.012) was significantly smaller in comparison to appropriate controls.As can be seen, there are no significant differences in the AH levels of the most abundant elements in comparison to the control groups for each patients’ group , whereas negative correlations (blue areas) or their absence (white areas) are much less frequent. Moreover, the clusters formed for the test groups, including patients with additional diseases such as AMD, diabetes, hypertension, and retinopathy, have greater strength in relation to the respective controls.+, K+) and alkali earth  ions by channels, carriers, and pumps has been studied for many decades. These ions as free or hydrated forms are the most abundant, and the mechanisms of their translocation are well known. It was established that the transport is governed mainly by electrochemical gradients supported with transmembrane protein transporters [+ transporters can also interact with Ag+, Au+, Pb2+ and Cd2+ [2+-ATPases are able to bind other metal ions such as Cu2+, Co2+, and Ni2+ [It is noteworthy that, apart from isolated pairs of elements, negative correlations mainly concerned sodium, and more rarely, potassium and magnesium. This observation appears to be very interesting in the context of metals homeostasis. The membrane transport of alkali  when compared with patients without AMD. This elevated cobalt level in the AH of patients with AMD was also observed previously by others [AMD is the progressive degeneration of the retinal pigment epithelium (RPE), retina, and choriocapillaris. This disease is observed among elderly people and is the leading cause of irreversible vision loss worldwide . The moland Mg2+  and hypoand Mg2+ . In our y others . Howeverp = 0.045) were reduced in the AH of patients diagnosed with hypertension when compared with patients without this disorder  in AH of patients suffering from diabetes and diabetic retinopathy was significantly smaller in comparison to appropriate controls . One migp = 0.014) similarly to P/Zn (p = 0.015).A much better parameter reflecting the metabolism of phosphorus in the body, which eliminates these individual differences, is the measurement of the P/Ca ratio. It was confirmed that the high concentration of serum phosphorus in diabetics is a result of renal insufficiency. When high levels of phosphorus are maintained for long periods, the driving force for mineralization appears and calcium phosphate may be deposited in abnormal sites , such as endothelial plaques in the arterial wall prosthetic, valvular, and extraskeletal calcifications ,58, and Without doubt, our study extends the scope of the metallomics research that has been limited to only a few selected elements so far. The strengths of this study also included a large control and study group and the consideration of the most common comorbidities as a variable. However, our study also had its limitations.First, we were not able to take into account other variables that potentially affect the levels of elements in body fluids, i.e., age, diet, and environmental exposure. Cataracts is a disease that usually appears late in life. Most patients undergoing cataract surgery are over 70 years of age. All patients came from one region of south-eastern Poland, so it can be roughly assumed that they were subjected to similar living conditions.Secondly, we did not verify other biological fluids or tissues that would allow us to evaluate the distribution of elements more broadly, as such studies were not covered by the approval of the Bioethics Committee.Thirdly, we had no influence on the possible contamination of the eye fluid during surgery, treatment, or possible supplementation. There was also no data to assess how comorbidities contribute to the progression of cataracts. Such analyses would require the long-term observation of selected groups of patients.The study population included patients attending the Department of General and Pediatric Ophthalmology at the Medical University of Lublin in Poland during the 2019–2021 period. The target population was cataract patients. The inclusion criteria covered the patients posted for cataract surgery in accordance with the applicable diagnostic and procedural codes developed by experts of the Polish Ophthalmology Society on 26 June 2018. Cataract patients diagnosed with additional comorbidities such as non-insulin-dependent diabetes mellitus (diabetes), hypertension, diabetic retinopathy, or age-related macular degeneration (AMD) were selected as cases, while the absence of certain comorbid diseases constituted controls. Patients with comorbidities received routine treatment appropriate to the disease using medicinal products prescribed “on-label” by attending physicians.The research material consisted of the fluid from the anterior chamber of the eye taken from patients at the beginning of the cataract extraction surgery. The cataract surgery was being performed in only one eye in each patient . The study was approved in 26 September 2019 by the Local Bioethical Committee of the Medical University of Lublin . The work was carried out in accordance with the Code of Ethics of the World Medical Association (Declaration of Helsinki) for experiments involving humans. The demographic characteristics of the patients are presented in 3 , followed by heating to 180 °C in closed Teflon containers in the microwave mineralization system Mars 6 . Finally, the samples were diluted to 10 mL by ultrapure water obtained in the purification system Milli-Q .The procedure was described previously . In brie−1, the auxiliary gas flow was 1.0 L min−1, the plasma gas flow was 12.0 L min−1, the charge-coupled device (CCD) temperature was −40 °C, the viewing height for the radial plasma observation was 8 mm, and the accusation time was 5 s. The analysis was repeated three times. ICP commercial analytical standards  were used for the calibration. The detection limits (LOD) were determined through 3-sigma criteria and were on the level of 0.00X mg/L (0.001–0.009 mg/L) wet weight (w/w) for all elements determined. The uncertainty  for the complete analytical process (including the sample preparation) was at the level of 20%. Due to lack of proper Certified Reference Material, the traceability was assessed by a standard addition procedure for a real sample. A recovery of 80–120% was considered acceptable for all the examined elements.The inductively coupled plasma optical emission spectrometer Agilent 5110 ICP-OES  was employed for elemental analysis. In doing so, the radio frequency (RF) power was 1.2 kW, the nebulizer gas flow was 0.7 L mint-Student test, was used for intergroup comparisons [The inter-elemental correlation analysis was conducted in the studied groups for all elements. All coefficients of Spearman correlation were clustered into homogenous groups within the control and experimental sample with hierarchical cluster analysis. To perform comparisons between groups in the context of element concentration, normality tests (Shapiro–Wilk test) and tests of homogeneity of variance (Levene test) were performed ,62,63. Tparisons ,65.To assess effect sizes, a biserial rank correlation coefficient was used, which takes values in the range , where (−1) means that values in the first population are smaller than values in the second population . ConversAll statistical analyses were performed in the R statistical environment (R Core Team 2016) , also usIn this study, AH, collected from patients undergoing cataract surgery was analyzed by ICP-OES for the presence of 67 elements. The patients had the most common cataract accompanying diseases such as AMD, retinopathy, hypertension, or diabetes. The performed statistical analysis revealed substantial variations in elemental composition between test groups in comparison to the control group with no co-existing morbidity. However, only a relatively small number of elements exhibited statistically significant variation. Levels of P and the ratio of P/Ca were different in retinopathy and diabetes. Levels of Co were significantly altered in AMD. In patients with hypertension, levels of Sn, Ti, and Ru varied significantly from the control group. Knowledge about the inter-correlations between elements in eye fluid may be useful in developing a strategy to combat cataracts, and planned future studies devoted to its pathogenesis.


This study investigated cross-cultural differences in individual pride and collective pride between Chinese and Americans using data from text corpora. We found higher absolute frequencies of pride items in the American corpus than in the Chinese corpus. Cross-cultural differences were found for relative frequencies of different types of pride, and some of them depended on the genre of the text corpora. For both blogs and news genres, Americans showed higher frequencies of individual pride items and lower frequencies of relational pride items than did their Chinese counterparts. Cross-cultural differences in national pride, however, depended on the genre: Chinese news genre included more national pride items than its American counterpart, but the opposite was true for the blog genre. We discuss the implications of these results in relation to the existing literature (based on surveys and laboratory-based experiments) on cultural differences in individual pride and collective pride. Pride is a basic human emotion and refers to a self-conscious emotion derived from one's achievements. Depending on whether such achievements are attributable to oneself or to a group to which one belongs, the sense of pride is labeled as either individual or group/collective pride  within the WEIRD samples and a much lower level of agreement among the non-WEIRD samples (r = 0.21). It is also worth noting that like Eid and Diener  and Beijing Language and the Culture University Corpus Center . The COCA contains more than 1.0 billion words of the English text (20 million words per year from 1990 to 2019) and is equally divided among spoken language, fiction, magazines, newspapers, academic texts, web pages, blogs, and TV/movies. The spoken genre mainly included conversations from TV and radio programs in the United States, and the fiction genre included short stories and plays from magazines. Magazines and newspaper genres were from nearly 100 different magazines and newspapers from across the United States. The academic genre included literature from peer-reviewed journals covering the range of science, social sciences, and humanities. Web pages included web genres of academic, argument, fiction, information, and so on, from the US portion of the GLoWbE corpus. Blogs included texts from the US portion of the GloWbE corpus that were classified by Google as blogs. TV/movies included subtitles from OpenSubtitles.org and later the TV and movies corpora.We used two freely available corpora in this study: the Corpus of Contemporary American English  extracted from news, spoken language , science and technology, literature, and other language styles. News genre materials were derived from the main newspapers in China, and the spoken genre mainly included information from microblogs. Science and technology genres included articles in academic journals published in China, and literature mainly included literary works.To summarize, both corpora are very large and widely used and should be considered as comprehensive data of language use in their respective countries Davies, . To imprIn the English corpus, we selected “pride” and “proud” along with their grammatical variations, including “prided,” “prides,” “priding,” “prouder,” and “proudest” , were divided into two groups: One group classified 854 English and 1,589 Chinese items, and the other group classified 2,141 English and 1,497 Chinese items. These four coders were trained how to classify the text by type of pride: individual pride, relational pride, national pride, general pride (the items with little or no information to be coded by the above types of pride), and non-pride  by standardizing the difference between observed values and expected values , than in the Chinese corpus (14.17 and 22.84 usages per million words).2 = 97.13, p < 0.001, Cramer's V = 0.18, which indicates slightly larger than a medium effect size (0.17) according to Cohen  = 278.50, p < 0.001, Cramer's V = 0.31, which is larger than a large effect size (0.29)  and pride , χto Cohen . For the) Cohen, . For bot) Cohen, . For natWe used the corpus-based methods to examine cultural differences in individual and collective  pride between Americans and Chinese. These methods can mitigate to some extent the clear methodological disadvantages of questionnaire-based methods, including ethnocentric, social desirability, and self-serving biases  included a higher proportion of individual pride items and a lower proportion of relational pride items than the Chinese corpus. These findings are consistent with our hypothesis derived from a theoretical discussion of the implications of individualism-collectivism and independent/interdependent self-construal of emotions  still play a significant propaganda-related role by focusing on the positive achievements of the country , it might have just reflected the consequences of the other cultural differences discussed above. To our knowledge, this study is the first to examine cultural differences in individual and collective pride based on specific corpora. Our findings extend the collectivistic characterization of Chinese to include relational pride and national pride. The results also support the notion that culture as well as other factors  plays an important role in emotional expression , to represent individualistic and collectivistic cultures. We did not include other major varieties of English language such as British and Australian English, which have been found to involve different cultural scripts in emotional expressions Goddard, . Future http://bcc.blcu.edu.cn/; https://www.englishcorpora.org/coca/.Publicly available datasets were analyzed in this study. This data can be found at: This study was carried out in accordance with recommendations of the Research Ethics Committee of Renmin University of China with written informed consent from all subjects. All subjects gave written informed consent in accordance with the Declaration of Helsinki. The protocol was approved by the Research Ethics Committee of Renmin University of China.CL and GY designed the original study and conducted final critical revision. JL, HW, and LY conducted literature searches and statistical analysis. CL and CC wrote and revised the manuscript. All authors contributed to the article and approved the submitted version.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


Ligusticum chuanxiong, including two transplants between the mountain and the dam area. It is well known that the environment can influence the endophytic community structure of plants. However, the change of host endophytic flora caused by transplanting in different places and its influence on asexual reproduction are still poorly understood. We carried out three cycles of cultivation experiments on L. chuanxiong and collected stem nodes (LZ), immature rhizomes (PX), medicinal rhizomes (CX), and rhizosphere. High-throughput sequencing was performed to analyze the endophytic fungi in all samples. We observed that the diversity and richness of endophytic fungi in L. chuanxiong increased as a result of transplanting cultivation from dam areas to mountains. Local transplantation caused minor changes in the endophytic fungus structure of L. chuanxiong, while remote transplantation caused significant changes. Compared with LZ after breeding in the dam area, the LZ after breeding on mountains has more abundant Gibberella, Phoma, Pericona, Paraphoma, and Neocosmospora. The regular pattern of the relative abundance of endophytic fungi is consistent with that of the fungus in the soil, while there are also some cases that the relative abundance of endophytic fungi is the opposite of that of soil fungi. In addition, there is a significant correlation among certain kinds of endophytic fungi whether in the soil or the plants. We have isolated more gibberellin-producing and auxin-producing fungi in the LZ cultivated in the mountains than that in the LZ cultivated in the dam area. The results of pot experiments showed that the three fungi isolated from LZ cultivated in mountainous areas can promote the development of shoots, stem nodes, and internodes of LZ, and increase the activity of plant peroxidase, catalase, phenylalanine ammonia lyase, and other enzymes. We can conclude that transplantation leads to the recombination of the host endophytic fungus, the more significant the difference in the environment is, the greater the reorganization caused by transplanting. Reorganization is determined by the soil environment, hosts, and the interaction of microorganisms. Remote transplantation is a crucial opportunity to reshuffle the micro-ecological structure of the asexual reproduction of plants, and regulate the growth, development, and resistance of plants, and prevent germplasm degradation caused by asexual reproduction.“Breeding on mountains, cultivation in dam areas” is a unique propagation method for the vegetatively propagated plant  In angiosperms, there are two pathways of reproduction: sexual and asexual (vegetative reproduction and apomixis) . VegetatLigusticum sinense, and Angelica sinensis can increase their respective yields by transplanting  is an effective medical plant, and has been extensively applied with other Chinese herbal medicines for many years to treat various diseases , the transplanting method of “breeding on mountains and cultivation in dam areas” has been adopted , Xiaoyudong (998 m ASL), and Taian (773 m ASL) ; all LZ The second part is a control experiment of breeding in mountains and breeding in the flat dam. Breeding was carried out at the same time in the mountain of Shuimo  and the dam area of Shiyang (592 m ASL). After harvesting, LZ was transplanted to Shiyang for medicinal rhizome cultivation. According to the breeding place, samples of LZ on the mountain and LZ in the dam area, CX samples, and corresponding soil samples were collected. The soils used in the two parts of the experiment were farming soil of local farmers. During the whole experiment, no fungicide, pesticide, or any form of fertilizer was applied .8 CFU and irrigated the PX that grew in a sterile substrate  with consistent growth. Meanwhile, a blank control group was established to ensure the validity of the results. Four months later, the growth index  and resistant enzyme content of L. chuanxiong were measured. We repeated 10 times for each group , and it was stored in an ultra-low temperature refrigerator at −80°C after being quickly frozen by liquid nitrogen . The rhizosphere soil was collected by shaking roots and was put into a sterilized 50-ml centrifuge tube after removing impurities and then quickly frozen in liquid nitrogen and kept at −80°C.TM-ST . Illumina paired-end sequencing libraries were prepared using the purified PCR products. After that, the libraries were sequenced using an Illumina MiSeq PE3000 platform. Negative control was established in each practical step to ensure the validity of the test results.Microbial genomic DNA was extracted from the samples using an HP Plant (Soil) Fungal Kit. The extract was detected by 0.8% agarose gel electrophoresis, and DNA concentration and purity were detected by a nucleic acid analyzer (Nanophotometer P330). We selected qualified samples on dry ice and sent them to Shanghai Meiji Company for high-throughput DNA sequencing . Fungal ITS-region (the intergenic transcribed spacer region between the 16S rRNA and 23S rRNA genes) was amplified using the forward primer ITS1F (5′-CTTGGTCATTTAGAGGAAGTAA-3′) and reverse primer ITS2R (5′-GCTGCGTTCTTCATCGATGC-3′) which are specific for fungi. The PCR reaction system included 4 μl of 10 × buffer, 2 μl of 2.5 mM dNTPs, 0.8 μl of forward and reverse primers, 0.2 μl of rTaq polymerase, 0.2 μl of bovine serum albumin (BSA), and 10 ng of DNA in a volume of 20 μl. Cycling parameters included initial denaturation at 95°C for 3 min, followed by 35 cycles of denaturation at 95°C for 30 s, annealing at 55°C for 30 s, elongation at 72°C for 45 s, and final elongation for 10 min at 72°C. PCR products were recovered by electrophoresis using 2% agarose gel. Subsequently, the recovered PCR products were purified using an AxyPrep DNA Gel Extraction Kit  and eluted with Tris–HCl. The concentration of the purified PCR products was checked by electrophoresis in 2% agarose gel and quantified using QuantiFluor2 for 15 min, and finally rinsed twice in sterile distilled water. Disinfected plant tissues were cut and macerated. The extract obtained was recovered and serial dilutions of 10–1, 10–2, and 10–3 were performed, and 100 μl of each dilution was plated on potato dextrose agar (PDA) . All plates were incubated at 28°C for 1 month. The plates were checked daily, and each emerging fungal colony was transferred onto a new PDA plate until axenic cultures were obtained.For fungal isolation, we took 2 g of fresh and healthy LZ for each part of the plant. The procedure was performed for the two plants independently. To isolate fungal epiphytes, tissues were rinsed with water and surface-sterilized in a sequence of 75% ethanol for 40 s followed by 2% NaClOPure fungal isolates were incubated in 5 ml of PDB medium, for 10 days at 30°C under dark conditions and agitation (150 rpm); 0.5 ml of culture media was recovered and combined with 0.5 ml of 96% ethanol and 5 ml of a cold mixture of sulfuric acid/ethanol (1:1 v/v). This mixture was incubated at 48°C for 30 min and then exposed to a UV lamp. Samples showing green fluorescence were considered positive for gibberellin production .4:0.5 mol/LFeCl3, 35:1 v/v) was added. This test tube was placed at room temperature and the light was avoided for 30 min. Samples showing pink fluorescence were considered positive for auxin (IAA) production , for 10 days at 30°C under dark conditions and agitation (150 rpm); 0.2 ml of the suspension was placed into a test tube, and an equal volume of Salkowski colorimetric solution , available phosphorus (AP), available potassium (AK), organic matter (OM), were analyzed according to standard soil-testing procedures . AccordiQ-value of less than 20, and unmatched barcode sequences) were removed. To obtain valid data, QIIME v1.9.0 is used for quality control, and the Uchime algorithm and gold database are used to remove delusion. These sequences were grouped into operational taxonomic units (OTUs) based on 97% sequence identity using UPARSE (V7.0.1090). Each row was annotated by comparing the Ribosomal Database Project (RDP) classifier (V2.11) against the SILVA (SSU123) database2 using a comparison threshold of 70%. Resampling was carried out with the smallest amount of data in the sample as the standard to make the uniform treatment for each sample. Mothur (version 1.30.2)1 is used for diversity analysis. Use R 3.6.0 was used to perform various data conversions. The ggplot2 package is used for population analysis, the stats package is used for Kruskal–Wallis rank sum test and cluster analysis, and the Vegan package is used to calculate Bray–Curtis distance and for PCA, RDA, and CCA analysis. FUNGuild  is used for function prediction. All the figures are made with ggplot2. Significant differences among groups were analyzed using one-way ANOVA, followed by Dunnett’s post-hoc test; p < 0.05 (marked as ∗) was defined as significant, p < 0.01 (marked as ∗∗) was defined as very significant, and p < 0.001 (marked as ∗∗∗ or more than ∗∗∗) was defined as extremely significant.Clean reads were obtained by filtering the raw sequences using a microbial ecological quantitative analysis pipeline . Low-quality sequences . The difference in the breeding environment is the most critical factor that causes the community structural reorganization of the endophytic fungi in L. chuanxiong.The results of the hierarchical cluster analysis show that the LZs, PXs, and CXs from Shuimo can be distinguished and that the samples at each stage tend to gather separately ; the LZsL. chuanxiong . However, the relative abundance of these genera at each growth stage fluctuates regularly  or decreased  in CX. In previous experiments, we also found a significant increase or and decrease, respectively, in CX in four other different origins.To learn more about the community structure of the endophytic fungi of The analysis results of endophytic fungi in CXs from four different producing areas in Sichuan province verified the reorganization of fungi, which indicates that the restructuring of endophytic fungi during transplanting will not disappear, regardless of the change in the production area.L. chuanxiong micro-ecological structure, we conducted two breeding experiments. In the breeding stage, we dug out PX and transplanted it to the dam area where the CX was cultivated for breeding, which is called “breeding in dam areas.” Simultaneously, we carried out traditional breeding on mountains and completed the follow-up cycle of medicinal material cultivation.To further explore the reasons for the formation of the All reads are over 200 bp, and the average length is about 252 bp, which is in line with expectations. The sequence coverage index Good’s Coverage extracted from each sample is greater than 99.9%. Microbial diversity was analyzed using the Shannon diversity index, and richness was analyzed using ace indices . The ShaL. chuanxiong at each stage under the M-Y cultivation mode accounted for 20.1%. The Y-Y cultivation mode accounted for 16.7% . There is more pathotroph in the samples from the mountain than the samples from the dam area, and pathotroph-saprotroph is just the opposite .For soils, there are more pathotroph and pathotroph-saprotroph in the soil of CX  than in the soil of LZ . Saprotroph is just the opposite, while pathotroph-saprotroph-symbiotroph has no difference in each sample of soil .Gibberella, Fusarium, Alternaria, Calophoma, Periconia, Didymella, Neocosmospora, Phoma, etc., and that the relative abundance in the samples from the M-Y cultivation mode is more significant than that in the samples from the Y-Y cultivation mode. The relationship is ML > YL > MY > YY. Where Gibberella, Didymella, Periconia, and Alternaria have a lower relative abundance in LZ’s soils of LZ and have a higher relative abundance in CX’s soils of CX (MYS > YYS > MLS > YLS) . In addition, regardless of the abundance of the soil, the relative abundance of Paraphoma and Calophoma in the samples of the M-Y cultivation model was significantly higher than that of the Y-Y cultivation model.We analyzed the difference in endophytic fungi in the samples of plants and soils under two transplanting modes . The resS > YLS) , while Fc fungus . CXs areL. chuanxiong by one-way ANOVA , Fusarium avenaceum (FAV), and Fusarium proliferatum (FP) , the FM, FAV, and FP strains could cause significant elongation of internodes and significant enlargement of the stem nodes, and that all the strains significantly increased the weight of the shoot of L. chuanxiong. In addition, FM can significantly increase the LZ coefficient (To investigate the impact of LZ core flora on the growth of tum (FP) . All thrfficient . There iL. chuanxiong, we measured the resistant enzyme content of the potted L. chuanxiong. The production of these resistance enzymes was quantitatively determined by a method described previously , while the relative abundance of Dactylonectria and Plectosphaerella is higher in the CX samples and its soil samples . It shows that the recombination of endophytic fungi caused by transplanting is brought by soil microorganisms to a certain extent.We found that the endophytic fungus of the plant is related to the soil. In the two cultivation modes of the control cultivating experiment, the diversity of endophytic fungi in the LZ samples was significantly higher than that in the CX samples in the same period, so were the corresponding soil samples. The relationship is MLS > MYS > ML > MY in the M-Y mode, and YLS > YYS > YL > YY in the Y-Y mode. In addition, the relative abundance of L. chuanxiong not transplanted has no significant changes. It fully shows that change in the breeding environment is the most critical factor for the reorganization of endophytic fungi in L. chuanxiong.Regardless of “breeding in dam areas ” or “breeding on mountains (remote transplanting),” as long as there is a transplanting process, it can lead to the reorganization of endophytic fungi. However, the endophytic fungal community structure of L. chuanxiong, always exist stably whether the plant is transplanting transplanted or not, which belong to the core fungal flora of L. chuanxiong and will not disappear because of different growth stages or cultivation areas. Soil is an essential factor in the composition of plant endophytic fungi. Its physical and chemical properties have a decisive effect on the microbial community structure in the rhizosphere and roots of plants  are the core flora of LZ, sodopathotroph-saprotroph-symbiotroph. The same result was verified in the control breeding experiment. In this study, it is not difficult to find that the relative abundances of the core flora of LZ, such as Gibberella, Alternaria, Periconia, Didymella, and Calophoma, are significantly higher in LZ than in CX; the opposite is true in the soil samples. Similarly, the relative abundance of pathotroph-saprotroph in the LZ on the mountain is significantly higher than in the dam area, but there is no significant difference in the corresponding two soils. The relative abundance of Plectosphaerella is significantly negatively correlated with Alternaria and Periconia in the plant samples. However, they are significantly positively correlated in the soil samples. By analyzing the correlation between these fungal communities and physiographical factors, we found that Gibberella, Didymella, Calophoma, Alternaria, and Periconia exhibited opposite correlations with environmental factors in the rhizosphere and endophytic state, which indicates that the characteristics of some plant endophytic flora tending to be similar with the growth and development of the host are not closely related to the soil, but are regulated by the growth law of the plant host.The soil-to-root transfer is one of the most thoroughly studied horizontal transmission routes for endophytes. It shows a great diversity of endophytic communities in the rhizosphere that are still far more diverse than those in the endosphere, and endophytic communities are often a subset of rhizosphere communities . In thisantation . On the uanxiong . At the Alternaria and Periconia have a significant positive correlation, so do Gibberella and Fusarium. Some Fusarium species have both asexual and sexual reproduction cycles . The coefficient of LZL. chuanxiong-resistant enzymes, such as CAT, POD, and PAL, indicating that they can all improve the defensive ability of L. chuanxiong. In the research results, we also found that the LZ cultivated in the mountainous area separated more gibberellin and auxin-producing strains than the LZ cultivated in the dam area, which indicates that there are indeed more growth-promoting fungi in the mountain environment.The pot experiment has proved the point. In a large body of literature, the three strains used for pot experiment are identified as pathogenic fungi, whether they were on stage  or Gibbeon stage , which cWe find that the relative abundance of some dominant genera in LZ will increase significantly as long as transplanting occurs. However, the relative abundance of these genera in transplanting on the mountain is significantly higher than in transplanting in the dam area and non-transplanting. Some of the dominant genera in CX only increase when transplanted in the mountains. Some genera increased when transplanted in mountainous areas, and the relative abundance does not change significantly when transplanted in the dam area or without transplanting.Regardless of breeding in mountains or breeding in the flat dam, recombination will occur as long as there is a transplanting process. The transplantation changes the chance of pathogen colonization in plant hosts, reduces the time for pathogens to interact with the host, and increases the colonization of beneficial or neutral endophytes. Plant pathogenic, beneficial, and neutral fungi have all obtained the opportunity to reshuffle during the recombination process. Remote transplantation brings more significant reorganization than local transplantation, and this reorganization is mainly reflected in the reduction of pathogenic fungi and the acquisition of core flora. It fully shows that remote transplantation is an important opportunity for reshuffling the micro-ecological structure of asexually propagated plants, which is an important process for the asexual reproduction of species to “rejuvenate.”2, and temperature)  . Abioticerature) . In the ic flora . On the ic flora . In thisThere is an intense chemical interaction between plants and microorganisms. In the rhizosphere, plants release root exudates, which promote microorganism population development. The rhizosphere offers varieties of carbon-rich micro-habitats, and beneficial microorganism populations can colonize by using such substrates . Plants Plant-associated microorganisms use chemotaxis to sense and respond to plant-derived signals, such as organic acids and sugars present in plant exudates. Once a signal is perceived, microorganisms move toward the plant to initiate colonization. Polyamines function as signaling molecules in the root–rhizosphere interface and inform the microbiome of the presence of eukaryotic hosts. Sensing of such molecules triggers a lifestyle switch to promote attachment and biofilm formation in many microbial groups . After sRubrivivax spp., as well as an enrichment of the type III secretion system (T3SS)-carrying Pseudomonas spp., relative to non-mycorrhizal plants  can be found below: NCBI BioProject. accession ZY: conceptualization. DH: methodology. GH, HL, and WX: resources. LK: writing—original draft preparation and writing—review and editing. DH, ZY, and LH: supervision, project administration, and funding acquisition. ZZ: participated in the experimental work. All authors have read and agreed to the published version of the manuscript.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.


The mechanism of ankylosing spondylitis with femoral head necrosis is unknown, and our study aimed investigate the effects of genetic and immune cell dysregulation on ankylosing spondylitis.The protein expression of all ligaments in ankylosing spondylitis with femoral head necrosis was obtained using label-free quantification protein park analysis of six pairs of specimens. The possible pathogenesis was explored using differential protein analysis, weighted gene co-expression network analysis, recording intersections with hypoxia-related genes, immune cell correlation analysis, and drug sensitivity analysis. Finally, routine blood test data from 502 AS and 162 healthy controls were collected to examine immune cell differential analysis.SAA1 and TUBA8 were significantly expressed differentially in these two groups and correlated quite strongly with macrophage M0 and resting mast cells (P < 0.05). Routine blood data showed that monocytes were significantly more expressed in AS than in healthy controls (P < 0.05). SAA1 and TUBA8 were closely related to the sensitivity of various drugs, which might lead to altered drug sensitivity.Dysregulation of SAA1, TUBA8 and monocytes are key factors in ankylosing spondylitis with femoral head necrosis. Ankylosing spondylitis (AS) is a chronic inflammatory disease, primarily involving the mid-axis skeleton. The main characteristic change is sacroiliac arthritis, and the disease occurrence is inextricably linked to genetics , 2. AS hHypoxia has been reported in several inflammatory diseases. Hypoxia is an important feature in pathological and physiological immune responses, and its effects on inflammation and immunity may vary depending on the specific immune processes and microenvironment. In pathological conditions, chronic inflammation, infection, and tissue ischemia, pathological hypoxia can lead to disease progression and tissue dysfunction through an imbalance in immune cell regulation . A studyMonocytes are a subtype of macrophage progenitors and dendritic cells whose primary role is to assume the role of circulating sensors for environmental changes and responses to disease . Recent Here, we searched for the role played by hypoxia-related genes in AS by performing label-free protein profiling of six ligamentous tissues from AS with femoral head necrosis and that without AS. Moreover, through a complex and precise bioinformatics approach, we searched for immune cells associated with AS in an attempt to find a new path for immunotherapy for AS. We also used data from routine blood tests to examine the results of our analysis. In addition, the drug sensitivity analysis was performed to provide a new basis of reference for the treatment of AS. The main objective of our study was to analyze the proteomic data of AS combined with femoral head necrosis and to find the genes closely associated with this disease through precise bioinformatics analysis, providing a new reference basis for the early diagnosis and treatment of this disease.We collected femoral head ligament tissues for pathological testing from patients who underwent an intraoperative artificial hip replacement at the First Clinical Affiliated Hospital of Guangxi Medical University between 2018 and 2019. This study was approved by the ethical review of the First Clinical Affiliated Hospital of Guangxi Medical University, as per the Declaration of Helsinki of the World Medical Congress. Inclusion criteria for this study were as follows: individuals with AS combined with femoral head necrosis were categorized into the experimental group, and those only with femoral head necrosis were categorized into the control group. Exclusion criteria for this study were: patients with combined rheumatoid arthritis, systemic lupus erythematosus, and tumors. A total of 12 patients were analyzed using a randomized method, with six patients in the experimental group and 6 in the control group. In our study, we used tissues from adults for the experiments and did not use tissues from minors for the experiments. Informed consent was obtained from all participants and/or their legal guardians.In our study, we quantified the proteins using the LFQ method to obtain accurate differences in the proteins of ligaments with AS combined with femoral head necrosis versus those only with femoral head necrosis. First, the samples were lysed, followed by mixing the radio-immune precipitation assay lysate with protease inhibitor and phenylmethylsulfonyl fluoride, which was placed on ice to pre-cool and obtain the working solution. Then the tissue samples were added to 1000 µL of working solution and mixed thoroughly, and sonicated in an ice bath for 5 min until they were fully dissolved. Subsequently, after centrifugation, a 96-well plate was added and shaken at 37°C. The protein concentration of the sample was calculated from the standard protein using the standard curve. Then, we performed acetone precipitation, protein redissolution, reduction, alkylation, enzymatic digestion, sodium deoxycholate removal, peptide desalting, nano-ultra performance liquid chromatography separation, plasmapheresis 120 min/sample, Maxquant analysis and label-free quantification, protein abundance distribution detection, and quantitative replication. All protein expressions of the experimental and control groups were obtained.In this study, we used the programming language R (version 4.0.2) to analyze the differentially expressed proteins to find the differentially expressed proteins in the experimental group versus the control group. We identified differentially expressed proteins between the two groups using the “ggplot2” package, the “limma” package, and the “pheatmap” package, with cut-off values. The cut-off value was set to |logFC| > 0.1, adjusted P < 0.05. Subsequently, we constructed a heat map of differentially expressed proteins, plotting the top 50 differentially significant up and down-regulated proteins. Also, we constructed volcano maps of differential proteins.unsigned =|cor|. The command R automatically selects the most suitable soft threshold β for the optimal slice. Finally, disease-related modules, as well as dynamic shear tree results, were obtained.WGCNA is an advanced bioinformatics analysis that is widely used in biological lineage information to find the biological modules most associated with the disease , 16. A gWe performed GO and KEGG pathway enrichment analyses for all differentially expressed proteins to gain insight into the function of these differential proteins between the AS and control groups. We used the “colorspace” package, “stringi” package, “ggplot2” package, “digest” package, “GOplot” package, “DOSE” package, “clusterProfiler” package, and “enrichplot” package to analyze the differentially expressed proteins. We set the screening conditions as P < 0.05 and adjusted-P < 0.05. Subsequently, the results of GO and KEGG enrichment analyses were obtained.https://www.string-db.org/) database to obtain the protein-protein interaction network, which was finally imported into Cytoscape (version 3.8.0) software for visualization.Here, all hypoxia-related datasets in humans were downloaded from the Gene Set Enrichment Analysis database to gain insight into the role of hypoxia-related genes in AS. Subsequently, intersections of the differentially expressed proteins obtained from the screen, the module with the highest disease relevance in WGCNA, and the hypoxia-related genes were taken, and the differentially expressed proteins with the highest disease relevance in AS were obtained. Finally, we imported all the differentially expressed proteins into the STRING . Subsequently, we used the programming language R  to analyze all protein gene expression information and drug sensitivity data using the “impute” , “limma”We performed a statistical analysis of monocytes, neutrophils, eosinophils and erythrocyte sedimentation rates in routine blood examinations of 502 AS and 162 normal controls from the First Clinical Affiliated Hospital of Guangxi Medical University in order to examine the differences in immune cells analyzed by bioinformatics. This study was approved by the Ethics Committee of the First Clinical Affiliated Hospital of Guangxi Medical University and was in accordance with the requirements of the Declaration of Helsinki of the World Medical Assembly.We randomly selected six ligament specimens from AS combined with femoral head necrosis and six from non-obligatory spondylitis with femoral head necrosis for LFQ protein park analysis. After sample lysis, BCA quantification, acetone precipitation, protein re-solubilization, reduction, alkylation and enzymatic digestion, peptide desalting, we obtained the expression of all proteins. As illustrated in the protein abundance plot  is a protein-coding gene, and the diseases most closely associated with this gene are amyloidosis Aa. Moreover, the results of the study by Lee et al. showed that in a non-pathogenic environment, cytokines from lymphoid cells produce interleukin (IL)-17. The immune cells such as T helper 17 (Th17) cells receive cytokines secreted by neighboring intestinal epithelial cells. SAAs are also capable of leading to a differentiation program of pathogenic pro-inflammatory Th17 cells and act directly with T cells in conjunction with signal transducer and activator of transcription 3-activating cytokines . SAA1 exTubulin Alpha 8 (TUBA8) is a protein-coding gene, and diseases closely related to this gene include polymicrogyria with optic nerve hypoplasia. TUBA8 is relatively high in mouse liver. TUBA8 is more strongly expressed in transformed cells compared to non-tumor tissues . In contHere, we examined the protein expression of hip ligaments with AS with femoral head necrosis as the experimental group and those with femoral head necrosis without AS as the control group using the label-free quantification protein park analysis laboratory for protein detection. The differentially expressed proteins of hypoxia were analyzed by a complex and precise bioinformatics approach to the differentially expressed proteins, the WGCNA approach to the modules most closely related to AS, and the operation of taking intersections of hypoxia-related genes. Furthermore, by analyzing the relationship between these proteins and immune cells, as well as the differential analysis of immune cells we found that monocytes/macrophages differed significantly in the AS and control groups. We examined the differentially expressed genes using two GEO datasets and the test results supported our analysis. We tested the immune cell differential analysis using routine blood data from 502 AS and 162 healthy controls, and the test results supported our analysis. Finally, we also found a strong association between TUBA2 and SAA1 and drug sensitivity of multiple drugs, which provides a basis for new insights into the use of drugs for the treatment of AS.This study had certain limitations as other studies. First, the inadequacy of the sample size. Considering the analysis of large samples, we only used six pairs of a total of 12 samples for the protein park test, which was insufficient. There were limitations in using routine blood data from 502 AS samples and 162 healthy control samples to examine differential immune cell analysis. Secondly, our study used a large number of laboratory manipulations and experiments for the detection of proteins in the early stage; thus, leaving a gap with the multifaceted and multilevel verification of our experimental results.Dysregulation of SAA1, TUBA8 and monocytes are key factors in ankylosing spondylitis with femoral head necrosis.The original contributions presented in the study are included in the article/The studies involving human participants were reviewed and approved by Ethics Department of the First Clinical Affiliated Hospital of Guangxi Medical University. Written informed consent for participation was not required for this study in accordance with the national legislation and the institutional requirements.JJ, CL, and XZ designed the study. TL, LC, SH, and XS analyze the data. WJ, JC, TC, HL, and YY digital visualization. SW and JZ collected data on routine blood data. JJ wrote and revised the manuscript. CL and XZ revised the manuscript. All authors read and approved the final manuscript. All co-authors participated in the laboratory operation. All authors read and approved the final manuscript.This study was supported by the Youth Science Foundation of Guangxi Medical University, Grant/Award Numbers: GXMUYFY201712; Guangxi Young and Middle-aged Teacher’s Basic Ability Promoting Project, Grant/Award Number: 2019KY0119.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.


Changes of immune cells in patients with hepatocellular carcinoma treated by radiofrequency ablation and hepatectomy, a pilot study. Open Life Sci. 2021;16(1):1002–9. The authors apologize to the Journal, readers, and their colleagues for the mistake and any inconvenience it caused.


Sleep is crucial for maintaining the recovery and restoration of the body and brain. Less sleep is associated with poor mental and physical performance. Seasonal changes in sleep patterns can be observed. This paper examines seasonal effects on sleep timing, duration, and problems in two Cree First Nation communities in Saskatchewan, Canada. Data were available from a community survey of 588 adults aged 18 years and older (range: 18–78 years) with 44.2% males and 55.8% females. Results are presented using descriptive statistics and a binary logistic-regression model to identify the association between seasonal changes in sleep patterns, and demographic, social, and environmental factors. The participants reported sleeping the least during the spring and summer months and sleeping the most during the fall and winter months. This was further confirmed by sleep hours and the lower proportion of recommended hours of sleep during the spring and summer, and a higher proportion of longer sleep duration during the fall and winter months. There was no significant variation in sleeping onset and wake-up times by season. Overall, there were no significant differences in the prevalence of sleep deprivation, insomnia, and excessive daytime sleepiness by season. When stratified by age group and sex, some differences existed in the prevalence of sleep problems by season. More than two-thirds (68.6%) of the participants reported that there was a change in sleep patterns across seasons, and about 26.0% reported a very or extremely marked change in sleep patterns across seasons. Changes in sleep patterns by season were related to money left at the end of the month and damage caused by dampness in the house. Studies showed that seasonal changes are associated with sleep duration and problems ,2,3,4. SSymptoms of depression and mood disorders are reported to vary with seasons ,11,12,13The data for this study came from a baseline survey undertaken during the First Nations Sleep Health Project (FNSHP) conducted in partnership with two Cree First Nation communities (Communities A and B) in Saskatchewan in 2018–2019. The purpose of the FNSHP was to examine the relationships between sleep disorders, and risk factors and comorbidities, and to evaluate local diagnosis and treatment. The study was approved by the University of Saskatchewan’s Biomedical Research Ethics Board (certificate no. Bio #18-110) and adhered to Chapter 9  in the Tri-Council Policy Statement: Ethical Conduct for Research Involving Humans [Trained research assistants from each community conducted the baseline surveys in their respective communities. Adults 18 years and older were invited to the community health and youth centre to complete the interviewer-administered questionnaires and clinical assessments. A pamphlet describing the study and an invitation to participate were distributed by the research assistants during local community events such as Treaty Days and during door-to-door canvassing. Simultaneously, there was a social-media campaign to invite the community members to participate in the survey. The survey collected information on demographic variables, individual and contextual determinants of sleep health, self-reported height and weight, and objective clinical measurements. This manuscript is based on data from the questionnaires. Demographic information about participants including age, sex, body-mass index, education level and money left at the end of the month, lifestyle factors, house environment, medical history, and sleep-health information was obtained from the survey questionnaire. The following questions addressed seasonal effects on sleep patterns using the modified Seasonal Pattern Assessment Questionnaire (SPAQ) : (i) “Atp values were reported using R function fisher.test, which used 2000 simulations. Sleep duration hours by season using median test and pairwise comparisons were compared using Mood’s median test [p < 0.20 were considered for the multivariate model. All variables that were statistically significant (p < 0.05), and important clinical factors  were retained in the final multivariable model. Interactions between potential effect modifiers were examined and retained in the final model if the p value was <0.05. The strengths of the associations were presented by odds ratios (OR) and their 95% confidence intervals (CI) [Statistical analyses were conducted using SPSS version 27  and R soian test ,29. Two ian test . Sleep cian test . These cals (CI) .A total of 588 participants completed the baseline survey: 418 individuals from Community A and 170 individuals from Community B. Demographic information about the participants was presented elsewhere and is only briefly described here ,33. MeanOf the participants, 77% (455/588) responded to questions related to time of the year during which they slept least and most . Accordip = 0.014), with a smaller proportion of respondents who woke up early in winter compared to the three other seasons , clinical insomnia (p = 0.370), or excessive daytime sleepiness (p = 0.148)). However, there was a higher prevalence of sleep deprivation (40.3%), insomnia (38.5%), and excessive daytime sleepiness (40.4%) in spring when compared to the three other seasons  and among males (p = 0.009) in the spring season. Males had significantly lower prevalence of sleep deprivation in the fall (p = 0.019). There was significantly low prevalence of insomnia among participants aged 60 years and older age group in the fall (p = 0.016), while excessive daytime sleepiness was significantly higher for this age group in the spring (p = 0.016) and significantly lower in the fall (p = 0.002).A significant variation in sleep problems was observed in the age and sex stratification . FurtherOf the participants, 360 responded to the question related to how much their sleep changes with the seasons. Of those, 68.6% reported a slight, moderate, very, or extremely marked change, with 10.3% reporting an extremely marked change. About one-third (31.4%) of the participants reported that there was no change in their sleep patterns across seasons . Sleep cThe bi- and multivariate-regression model results are presented in In this study, participants reported sleeping the least during the months of May–August, and sleeping the most during the months of November–February. This was further confirmed by sleep hours and the lower proportion of recommended hours of sleep during the spring and summer, and a higher proportion of longer duration of sleep during the fall and winter months. There was no reported significant variation of sleeping onset and wake-up times by season. Reported hours of sleep by seasons confirmed that there was a significant difference in sleep hours during spring and summer. Overall, there were no significant differences in the prevalence of sleep deprivation, insomnia, and EDS by season. When stratified by age group and sex, some differences existed in the prevalence of sleep problems by season. More than two-thirds of participants reported that there was a change in sleep patterns across seasons, and about 10% reported an extremely marked change. Changes in sleep patterns by season were related to age, money left at the end of the month, damage caused by dampness in house, and having access to a nurse.Seasonal changes in sleep problems were studied in many countries ,35,36,37Several studies reported that there were seasonal changes in sleep onset and offset ,42,43,44Numerous studies reported seasonal variation in sleep duration ,4,46. HaIn this study, 26.4% reported a very or extremely marked change in sleep patterns across the seasons, and about 10% reported an extremely marked change. There were no significant associations between age group and changes in sleep patterns by season. In contrast, Suzuki et al. [Some authors reported an association between socioeconomic status and sleep issues ,48. LallHowever, this study did not observe change in sleep patterns with participants who had access to any healthcare providers. Nurses are often the communities’ main point of contact with the healthcare system . In SaskData on sleep onset and wakeup times, sleep duration, seasonal variation, and sleep problems were based on self-reporting, which could have biased the findings. Sleep duration was not assessed separately for work or school and free days, or for weekends or holidays. The difference between seasons in the number of working days may have confounded the results. Many other factors, including psychiatric diagnoses, physical comorbidities, the use of sedatives, and other lifestyle factors, were not assessed. We do not have repeated data for participants for all four seasons. Therefore, we were not able to explore the individual changes by season, but could only identify population-level changes by seasons. Due to the small sample size of changes by season outcome, we were unable to see whether there were group differences according to when people sleep longer or shorter, and whether the same factors of changes by season hold in different sleep-duration groups. This study reported seasonal changes in sleep patterns in two Saskatchewan First Nation communities. These changes in sleep patterns were influenced by age, income status, dampness in house, and access to a nurse. This information is helpful for practice improvement by healthcare providers in treating and managing patients with sleep disorders, policy development, and future research priorities in and with these communities.