Deciphering spatio-seasonal patterns, driving forces, and human health risks of nitrate and fluoride enriched water bodies in the Inner Mongolia Reaches of the Yellow River Basin, China.

The ecology and environment of the Yellow River Basin is threatened by fluoride and nitrate contamination induced by anthropogenic activity and geogenic factors. As a result, deciphering the spatio-temporal variability of fluoride and nitrate contamination in this area remains a challenge. Three hundred eighty-six samples of surface water and groundwater from the Inner Mongolia Reaches of the Yellow River Basin were taken for this investigation. According to the results of the multivariate statistical and geostatistical analyses, the fluoride pollution was primarily discovered in the middle and lower reaches of the study area and was determined to be more severe during the dry season. In contrast, nitrate contamination was found to be more severe during the wet season while being widely distributed in groundwater and concentrated in areas with intensive agricultural activities. The primary mechanisms governing the spatial-seasonal patterns of NO3- and F- pollution were shown by the principal component analysis, isotopic, and hydrochemical diagrams. The water-rock interaction or evaporation was crucial in the enrichment of F-. The human inputs (e.g., fertilizer or sewage) dominated fluoride and nitrate contamination. Additionally, the alkaline environment played a role in the generation of NO3- and F-. The health risk assessment concluded that the threat of fluoride contamination was greater than that of nitrate contamination. Children faced the greatest health risks, followed by females and males. These findings would serve as a guide for water management and pollution control in the Yellow River Basin.

[Water Chemical Isotope Characteristics and Water Transformation Relationship in Mongolian Section of the Yellow River Basin].

The Yellow River in Inner Mongolia was selected as the study area in this study. In July (wet season) and October (dry season) of 2021, the acquisition of seasonal rivers, the Yellow River tributaries and precipitation, the Yellow River, Wuliangsuhai, Lake Hasuhai, Lake Daihai, an irrigation canal system, and underground water and sea water samples were collected to test the water chemical composition and hydrogen and oxygen isotopic values of different water types. Using the Piper triplot, Gibbs plot, ion ratio, and MixSIAR model methods, the evolution of water chemistry in the Mongolian section of the Yellow River Basin was analyzed, and the transformation relationship between precipitation, surface water, and groundwater was revealed. The results showed that both groundwater and surface water in the study area were slightly alkaline; the dominant anion in water was Cl-, and the dominant cation was Na+. The main hydrochemical types of surface water were Cl·SO4-Na·Mg and SO4·HCO3-Na·Mg, whereas those of groundwater were Cl·SO4-Na·Mg and SO4·HCO3-Na·Ca. Groundwater Ca2+ and Mg2+ were primarily derived from the dissolution of silicate and evaporite, and surface water Ca2+ and Mg2+ were primarily derived from carbonate karst dissolution and carbonate and sulfuric acid in water participating in the dissolution process of carbonate and sulfide minerals. Na+ and Cl- in different water bodies were all affected by anthropogenic pollution sources. Owing to the seasonal effect, δD and δ18O of surface water and groundwater were higher in the wet season than in the dry season. The results showed that surface water was affected by evaporative fractionation after receiving precipitation recharge, and the groundwater recharge sources were complex. The MixSIAR model revealed that surface water was the main recharge source of groundwater, accounting for 52.4%-62.2% of the total recharge, and atmospheric precipitation was the main recharge source of surface water, accounting for 85.4%-97.1% of the total recharge.

SPRY4-dependent ERK negative feedback demarcates functional adult stem cells in the male mouse germline†.

Niche-derived growth factors support self-renewal of mouse spermatogonial stem and progenitor cells through ERK MAPK signaling and other pathways. At the same time, dysregulated growth factor-dependent signaling has been associated with loss of stem cell activity and aberrant differentiation. We hypothesized that growth factor signaling through the ERK MAPK pathway in spermatogonial stem cells is tightly regulated within a narrow range through distinct intracellular negative feedback regulators. Evaluation of candidate extracellular signal-regulated kinase (ERK) mitogen-activated protein kinase (MAPK)-responsive genes known to dampen downstream signaling revealed robust induction of specific negative feedback regulators, including Spry4, in cultured mouse spermatogonial stem cells in response to glial cell line-derived neurotrophic factor or fibroblast growth factor 2. Undifferentiated spermatogonia in vivo exhibited high levels of Spry4 mRNA. Quantitative single-cell analysis of ERK MAPK signaling in spermatogonial stem cell cultures revealed both dynamic signaling patterns in response to growth factors and disruption of such effects when Spry4 was ablated, due to dysregulation of ERK MAPK downstream of RAS. Whereas negative feedback regulator expression decreased during differentiation, loss of Spry4 shifted cell fate toward early differentiation with concomitant loss of stem cell activity. Finally, a mouse Spry4 reporter line revealed that the adult spermatogonial stem cell population in vivo is demarcated by strong Spry4 promoter activity. Collectively, our data suggest that negative feedback-dependent regulation of ERK MAPK is critical for preservation of spermatogonial stem cell fate within the mammalian testis.

Viral Transduction of Mammalian Spermatogonial Stem Cells.

Lentiviral vectors have been major tools for genetic manipulation of spermatogonial stem cells (SSCs) in vitro. Adeno-associated viral vectors are promising emerging tools for in vivo SSC transduction that are less invasive, compared to lentivirus, since AAV DNA is not integrated into the host genome and the host genome remains intact. In this chapter, we describe protocols using lentiviral and adeno-associated viral vectors to transduce SSCs in vitro and vivo, respectively.

Dynamic characteristics and synergistic effects of ecosystem services under climate change scenarios on the Qinghai-Tibet Plateau.

The Qinghai-Tibet Plateau (QTP) supplies many ecosystem services (ESs) that maintain local and global pan-Asian populations and ecosystems. The effects of climate change on ES provision in the QTP will have far-reaching impacts on the region and the many downstream ecosystems and countries that depend on ESs from the "Third Pole". This study undertook a systematic assessment of ES provision, trade-offs and synergies between four ESs (raw material provision, water yield, soil retention, and carbon storage) under future climate scenarios (representative concentration pathway). The results show that: (1) the total amount of the four ESs on the QTP is predicted to increase from 1980 to 2100 for three climate change scenarios. (2) The spatial pattern of ESs on the QTP will not change significantly in the future, and the grassland and forest ESs in the central and southern regions are predicted to increase significantly. (3) The synergistic interactions among ESs were generally consistent at three spatial scales (10 km (pixel), county and watershed scales), but with more significant synergistic effects at the watershed scale. This demonstrates the necessity for the examination of scale-dependent ES dynamics and interactions. This study will supply a reference for further research on long-term ES assessments, especially the dynamic ES changes and the spatial scale dependency of the ES interactions, and provide evidence-based strategies for formulating ecosystem management on the QTP under climate change.

Research on a Novel Locating Method for Track Inspection Based on Onboard Sensors in Maglev Train.

Guideway inspection is of great significance to the operation safety and riding quality of a commercial high-speed maglev transportation system. When analyzing guideway inspection data, it is important to obtain the location information for each piece of raw data and convert it from the time domain to the spatial domain for the analysis afterward. Previous studies have used the method of adding additional hardware such as GPS (global positioning system) receivers, LRF (location reference flag) readers, or onboard CAN (controller area network) bus adaptors to obtain location information. This paper presents a novel method for indirectly obtaining the location information via the use of data from the levitation and guidance control sensors perpendicular to the longitudinal direction to extract the characteristic information from the track. The method can be used for a long stator linear motor-driven maglev system and similar contactless rail transit systems. The results showed that the method could accurately identify the required location information in each stator tooth during the entire operation simultaneously with the operating information such as train velocity, direction, and track ID, without additional hardware installation and vehicle network connection. Thus, it could improve the pertinence of the results of guideway inspection equipment, and at the same time, facilitate the miniaturization and independence of guideway inspection equipment.

Panoramic transcriptome analysis and functional screening of long noncoding RNAs in mouse spermatogenesis.

Long noncoding RNAs (lncRNAs) have emerged as diverse functional regulators involved in mammalian development; however, large-scale functional investigation of lncRNAs in mammalian spermatogenesis in vivo is lacking. Here, we delineated the global lncRNA expression landscape in mouse spermatogenesis and identified 968 germ cell signature lncRNAs. By combining bioinformatics and functional screening, we identified three functional lncRNAs (Gm4665, 1700027A15Rik, and 1700052I22Rik) that directly influence spermatogenesis in vivo. Knocking down Gm4665 hampered the development of round spermatids into elongating spermatids and disrupted key spermatogenic gene expression. Mechanistically, lncRNA Gm4665 localized in the nucleus of round spermatids and occupied the genomic regulatory region of important spermatogenic genes including Ip6k1 and Akap3 These findings provide a valuable resource and framework for future functional analysis of lncRNAs in spermatogenesis and their potential roles in other biological processes.

Study on the osmoregulation of "Halomonas socia" NY-011 and the degradation of organic pollutants in the saline environment.

"Halomonas socia" NY-011, a new species of moderately halophilic bacteria isolated and identified in our laboratory, can grow in high concentrations of salt ranging from 0.5 to 25%. In this study, the whole genome of NY-011 was sequenced and a detailed analysis of the genomic features was provided. Especially, a series of genes related to salt tolerance and involved in xenobiotics biodegradation were annotated by COG, GO and KEGG analyses. Subsequently, RNA-Seq-based transcriptome analysis was applied to explore the osmotic regulation of NY-011 subjected to high salt stress for different times (0 h, 1 h, 3 h, 6 h, 11 h, 15 h). And we found that the genes related to osmoregulation including excluding Na+ and accumulating K+ as well as the synthesis of compatible solutes (alanine, glutamate, ectoine, hydroxyectoine and glycine betaine) were up-regulated, while the genes involved in the degradation of organic compounds were basically down-regulated during the whole process. Specifically, the expression trend of genes related to osmoregulation increased firstly then dropped, which was almost opposite to that of degrading organic pollutants genes. With the prolongation of osmotic up-shock, NY-011 survived and gradually adapted to osmotic stress, the above-mentioned two classes of genes slowly returned to normal expression level. Then, the scanning electron microscope (SEM) and transmission electron microscope (TEM) were also utilized to observe morphological properties of NY-011 under hypersaline stress, and our findings showed that the cell length of NY-011 became longer under osmotic stress, at the same time, polyhydroxyalkanoates (PHAs) were synthesized in the cells. Besides, physiological experiments confirmed that NY-011 could degrade organic compounds in a high salt environment. These data not only provide valuable insights into the mechanism of osmotic regulation of NY-011; but also make it possible for NY-011 to be exploited for biotechnological applications such as degrading organic pollutants in a hypersaline environment.

A long noncoding RNA binding to QKI-5 regulates germ cell apoptosis via p38 MAPK signaling pathway.

Spermatogenesis is the complex process of male germline development and requires coordinated interactions by multiple gene products that undergo strict developmental regulations. Increasing evidence has suggested that a number of long noncoding RNAs (lncRNAs) may function as important regulatory molecules in various physiological and pathological processes by binding to specific proteins. Here, we identified a subset of QKI-5-binding lncRNAs in the mouse testis through the integrated analyses of RNA immunoprecipitation (RIP)-microarray and biological verification. Among the lncRNAs, we revealed that NONMMUT074098.2 (Lnc10), which was highly expressed in the spermatogonia and spermatocytes of the testis, interacted with QKI-5. Furthermore, Lnc10 depletion promoted germ cell apoptosis via the activation of p38 MAPK, whereas the simultaneous knockdown of QKI-5 could rescue the apoptotic phenotype and the activation of p38 MAPK, which were induced by the loss of Lnc10. These data indicated that the Lnc10-QKI-5 interaction was associated with the regulatory roles of QKI-5 and that the Lnc10-QKI-5 interaction inhibited the regulation of QKI-5 on the downstream p38 MAPK signaling pathway. Additionally, we functionally characterized the biological roles of Lnc10 and found that the knockdown of Lnc10 promoted the apoptosis of spermatogenic cells in vivo; this suggested that Lnc10 had an important biological role in mouse spermatogenesis. Thus, our study provides a potential strategy to investigate the biological significance of lncRNA-RBP interactions during male germline development.

Rnf138 deficiency promotes apoptosis of spermatogonia in juvenile male mice.

Spermatogenesis, the process by which haploid sperm cells are produced from a diploid precursor cell, is essential for sexual reproduction. Here, we report that RING-finger protein 138 (Rnf138) is highly expressed in testes, especially in spermatogonia and spermatocytes. The role of Rnf138 in spermatogenesis was examined using a Rnf138-knockout mouse model. Rnf138 deficiency resulted in increased apoptosis in spermatogenic cells, loss of proliferative spermatogonia, delayed development of spermatozoa and impaired fertility. The proportion of PLZF+Ki67+ cells within the PLZF+ population decreased in the knockout mice. The phenotype was further assessed by RNA-sequencing (RNA-seq), which determined that the expression levels of many genes involved in spermatogenesis were altered in the testis of Rnf138-knockout mice. Thus, Rnf138 deficiency promotes the apoptosis of spermatogenic cells, which may have been caused by the aberrant proliferation of spermatogonia in mouse testis development.

[Expression Patterns of the Proteins Associated with Cell Junctions in Mouse Testes].

OBJECTIVE: To study on the expression patterns of proteins associated with cell junctions in the developing mouse testes. METHOD: The expression levels of reproductive related cell lines spermatogonia cell line GC1 spg, spermatocyte cell line GC2 spg, leydig cell line TM3, and sertoli cell line TM4, primary sertoli cells, and 1-6-week mouse testes were analyzed using Western blot. RESULTS: The sertoli cell junction-associated membrane proteins adhesion molecule A, Occludin and Claudin, and the sertoli-germ cell junction-associated membrane proteins junctional adhesion molecule C, Nectin-3, and E-cadherin were stage-specific in the seminiferous tubules in the mouse testes. The adaptor proteins associated with cell juctions zonula occludens-1, zonula occludens-2, Afadin, Β-catenin, and CD2-associated protein were not stage-specific in the seminiferous tubules in the mouse testes. CONCLUSIONS: In the seminiferous tubules in the mouse testes, the membrane proteins associated with cell junctions are stage-specific. However, the expressions of adaptor proteins associated with cell junctions do not obviously change.

Immune Infertility Should Be Positively Diagnosed Using an Accurate Method by Monitoring the Level of Anti-ACTL7a Antibody.

Infertility is currently a major public health problem. Anti-sperm antibodies (ASAs) markedly reduce sperm quality, which can subsequently lead to male and/or female infertility. The accurate detection of ASAs derived from specific spermatozoa is, therefore, clinically useful. We have focused on the spermatozoa-specific expression protein ACTL7a for many years and have developed an enzyme-linked immunosorbent assay (ELISA) to detect the concentration of anti-ACTL7a antibodies in fertile sera (n = 267) and infertile sera (n = 193). Infertile sera were collected from the positive sera of tray agglutination tests (TAT), which is a routine ASA screening methodology. We found that the concentration of anti-ACTL7a antibodies was significantly higher in the infertile sera (than in the fertile sera, P < 0.0001) and much higher in the TAT ≥ 16 infertile sera. The ELISA was much better for male sera detection (AUC = 0.9899). If we set the standard at a strongly positive value (calculated by ROC curve), the positive predictive value of the antibody detection reached 100 percent, with a false positive rate of zero. The developed ELISA method for anti-ACTL7a antibody detection is therefore sensitive, accurate, and easy to perform, making it an excellent potential tool for future clinical use.

Anti-GAPDHS antibodies: a biomarker of immune infertility.

Numerous investigations have focused on the detection of antisperm antibodies, which have a naturally occurring impact on male and female fertility. In this study, spermatogenic glyceraldehyde-3-phosphate dehydrogenase (GAPDHS) was considered to be a candidate biomarker of immune infertility. The concentrations of anti-GAPDHS antibodies in the sera of sterile individuals and fertile couples were measured by enzyme-linked immunosorbent assay. Sera were collected from immune infertile (n = 175) and fertile (n = 237) individuals and were screened by tray agglutination tests (TAT). Infertile sera were further divided into two groups according to the serum titers obtained by TAT (titers ≤ 1:8, n = 58; titers > 1:8, n = 117). The concentrations of anti-GAPDHS antibodies were significantly higher in the immune infertile group than in the fertile group and were much higher with regard to the increased degrees of sperm agglutination (titers > 1:8). Surprisingly, we found statistically significantly higher concentrations of antibodies in the sera of infertile men than in those of infertile women, and a similar statistical result was obtained in the sera when primary infertility was compared with secondary infertility. Thus, anti-GAPDHS antibodies seem to be a sensitive parameter in immune infertile detection and might be one of the main factors causing immune infertility. This factor might be valuable as an indicator in the clinical diagnosis and monitoring treatment of infertility.

A novel mutation of EYA4 in a large Chinese family with autosomal dominant middle-frequency sensorineural hearing loss by targeted exome sequencing.

The middle-frequency sensorineural hearing loss (MFSNHL) is rare among hereditary non-syndromic hearing loss. To date, only three genes are reported to be associated with MFSNHL, including TECTA, EYA4 and COL11A2. In this report, we analyzed and explored the clinical audiological characteristics and the causative gene of a Chinese family named HG-Z087 with non-syndromic autosomal dominant inherited MFSNHL. Clinical audiological characteristics and inheritance pattern of a family were evaluated, and pedigree was drawn based on medical history investigation. Our results showed that the Chinese family was characterized by late onset, progressive, non-sydromic autosomal dominant MFSNHL. Targeted exome sequencing, conducted using DNA samples of an affected member in this family, revealed a novel heterozygous missense mutation c.1643C>G in exon 18 of EYA4, causing amino-acid (aa) substitution Arg for Thr at a conserved position aa-548. The p.T548R mutation related to hearing loss in the selected Chinese family was validated by Sanger sequencing. However, the mutation was absent in control group containing 100 DNA samples from normal Chinese families. In conclusion, we identified the pathogenic gene and found that the novel missense mutation c.1643C>G (p.T548R) in EYA4 might have caused autosomal dominant non-syndromic hearing impairment in the selected Chinese family.

Regulatory effect of microRNA-135a on the Th1/Th2 imbalance in a murine model of allergic rhinitis.

Allergic rhinitis (AR) is primarily caused by a T helper cell (Th)1/Th2 imbalance. In a murine AR model of a previous study, the serum ovalbumin (OVA)-sIgE concentration was high, whereas microRNA (miR)-135a was lowly expressed in the nasal mucosa. The abnormal expression pattern of miR-135a coincided with highly expressed endogenous factors, including GATA binding protein (GATA)-3 and interleukin (IL)-4, and lowly expressed factors, including T-box expressed in T cells (T-bet) and interferon (IFN)-γ. We hypothesized that miR-135a may play an important role in immune regulation in AR mice. In the present study, AR was induced by OVA in the mice. Two groups of the AR mice were treated with a miR-135a mimic and a mimic control, respectively. The serum and nasal mucosa were collected for analysis. Following miR-135a application, the serum OVA-sIgE concentration was significantly reduced. In the nasal mucosa, the expression levels of miR-135a were higher, the mRNA and protein expression levels of GATA-3 and IL-4 were lower, and the expression levels of T-bet and IFN-γ were higher. The miR-135a corrected the Th1/Th2 imbalance in the AR mice. Findings of this study may provide a basis for novel genetic treatments in addressing allergic diseases.

Paradoxical enhancement of active cochlear mechanics in long-term administration of salicylate.

Aspirin (salicylate) is a common drug and frequently used long term in the clinic. It has been well documented that salicylate can cause reversible hearing loss and tinnitus and diminish outer hair cell (OHC) electromotility, which is capable of actively boosting the basilar membrane vibration and producing acoustic emission. However, aspirin's ototoxic mechanisms still remain largely unclear. In this experiment, the effects of long-term salicylate administration on cochlear hearing functions were investigated by measuring distortion product otoacoustic emissions (DPOAEs) in awake guinea pigs. A single injection of sodium salicylate (200 mg/kg) could reduce the amplitude of the cubic distortion product of 2f1-f2 within 2 h. The reduction was significant at 20-50 dB SPL stimulus levels and recovered after 8 h. However, following daily injections of sodium salicylate (200 mg/kg, b.i.d.), the distortion product of 2f1-f2 progressively increased. After injection for 14 days, the distortion product increased about 2-3.5 dB SPL. The increase rate was about 0.2 dB SPL/day. The DP-I/O function remained nonlinear. The increase was greater at 40-70 dB SPL primary sound intensities and reversible. After cessation of salicylate treatment for 4 wk, the increased distortion product returned to the initial normal levels. The rate of recovery was 0.1 dB SPL/day. In the control animals with saline injection, there was no change in DPOAEs. The data revealed that long-term administration of salicylate could paradoxically enhance active cochlear mechanics. The data also suggested that salicylate-induced tinnitus might be generated at the OHC level.

[The changes of some vasoactive substances after operation in patients with obstructive sleep apnea-hypopnea syndrome].

OBJECTIVE: To observe the changes of thromboxane B2 (TXB2), rennin, angiotensin II (AT-II), endothelin 1 (ET-1) and anticardiolipin antibody (ACA) in patients with obstructive sleep apnea-hypopnea syndrome (OSAHS) before and after the operation. METHOD: Fifty-four cases of OSAHS confirmed by polysomnography (PSG) were selected as the OSAHS group and 20 normal donors without OSAHS were selected as the control group. Plasma levels of TXB2, rennin, AT- II, ET-1 and ACA were detected by enzyme-linked immunosorbent assay (ELISA). RESULT: Plasma levels of TXB2, rennin, AT-II, ET-1 and ACA were higher in patients with OSAHS than those in control group (P < 0.05 or P < 0.01), and the operation therapy decreased them significantly (P < 0.05 or P <0.01). All the plasma parameters were correlated positively with AHI, and negatively with SaO2. CONCLUSION: The results indicate the patients with OSAHS were susceptible to thromboembolism disease. Operation therapy is effective in correcting TXB2, rennin, AT-II, ET-1 and ACA.

[Clinical analysis of selected transarterial embolization in treating uncontrolled epistaxis].

OBJECTIVE: To learn the effects and the correlative factors of selected transarterial embolization in treatment of uncontrolled epistaxis. METHOD: The abnormal ends of internal maxillary artery were selected embolized in 23 patient of uncontrolled epistaxis by digital subtraction angiography (DSA) of carotid, meanwhile the facial artery were also embolized in 5 patients. RESULT: It was confirmed that hypogenesis and malformation in the ends of internal maxillary artery were existed in all 23 cases during DSA. There was evidence of bleeding in 15 patients, and facial artery with abnormal shape was found in 6 patients. Nose-bleeding was stopped in 22 cases (95.6%) after embolized, including one had a recurrence of bleeding in 24 hours because of the foundation of new abnormal communications. The fail in one case might be related with taking out all the stuff in noses before embolization and/or bleeding facial artery. CONCLUSION: Uncontrolled epistaxis in this report might be caused by hypogenesis and malformation of internal maxillary artery. The effects of selected transarterial embolization were rapid and satisfactory. The fail in treatment might be related to foundation of new abnormal communications and bleed from facial artery. Taking out all the stuff in noses too early might also influence the effects of embolization.