Sustainability of new energy vehicles from a battery recycling perspective: A bibliometric analysis.

In recent years, new energy vehicles (NEVs) have taken the world by storm. A large number of NEV batteries have been scrapped, and research on NEV battery recycling is important for promoting the sustainable development of NEVs. Battery recycling is an important aspect of the sustainable development of NEVs. In this study, we conducted an in-depth analysis of the current status of research on NEV battery recycling from a new perspective using bibliometric methods and visualization software. This study shows that research targeting the recycling of NEV batteries is growing rapidly, and collaborative networks exist among researchers from different countries, institutions, and fields. The focus of research has shifted from lead-acid batteries to lithium batteries, and the supply chain and circular economy related to NEV battery recycling is an emerging research hotspot. Based on our analysis, we propose that the government should establish policies to improve the recycling networks at the collection stage and provide subsidies to attract consumers. Enterprises should develop low-cobalt and cobalt-free technologies, utilize green solvents, and develop new battery swap modes. The establishment of an information platform is conducive to the further development of collaborative networks.

Physical activity and school adaptation among Chinese junior high school students: chain mediation of resilience and coping styles.

Background: As a crucial juncture in students' educational journey, junior high school presents challenges that profoundly influence well-being and academic performance. Physical activity emerges as a pivotal factor shaping the holistic development of junior high school students. Beyond its recognized impact on physical and mental health, engaging in regular physical activity proves effective in augmenting students' adaptability to school life. Despite its importance, the mechanisms through which physical activity influences school adaptation in junior high school students remain understudied in academic research. Objective: In exploring the potential mechanisms, this study aims to validate the mediating roles of resilience and coping styles by examining the association between physical activity and school adaptation among junior high school students. Methods: This study employed cross-sectional survey approach among junior high school students in China. Through the convenience sampling, 1,488 participants aged from 12 to 16 years old (Average age = 13.59, SD = 1.017) from two Junior high schools in Changsha City, Hunan Province were recruited to complete the Physical Activity Scale, School Adaptation Questionnaire for Junior High School Students, Resilience Scale for Adolescents, and Simple Coping Styles Questionnaire. For data analysis, the SPSS 26.0 and Amos 26.0 were used for statistical processing. Results: The results showed that physical activity exhibited a significant correlation with school adaptation (r = 0.656, p < 0.001). Resilience, positive coping style and negative coping style played partial mediating roles between physical activity and school adaptation, with the effect size were 0.229, 0.170, 0.171. The chain mediation effect size of resilience and positive coping style was 0.042, while the chain mediation effect size of resilience and negative coping style was 0.050. Conclusion: Physical activity positively predicts Chinese junior high school students' school adaptation through resilience and coping styles, suggesting that junior high school students should engage in regular physical activity, so as to improve their resilience and positive coping styles, mitigating negative coping styles, thus promoting their school adaptation.

Phage selection drives resistance-virulence trade-offs in Ralstonia solanacearum plant-pathogenic bacterium irrespective of the growth temperature.

While temperature has been shown to affect the survival and growth of bacteria and their phage parasites, it is unclear if trade-offs between phage resistance and other bacterial traits depend on the temperature. Here, we experimentally compared the evolution of phage resistance-virulence trade-offs and underlying molecular mechanisms in phytopathogenic Ralstonia solanacearum bacterium at 25 °C and 35 °C temperature environments. We found that while phages reduced R. solanacearum densities relatively more at 25 °C, no difference in the final level of phage resistance was observed between temperature treatments. Instead, small colony variants (SCVs) with increased growth rate and mutations in the quorum-sensing (QS) signaling receptor gene, phcS, evolved in both temperature treatments. Interestingly, SCVs were also phage-resistant and reached higher frequencies in the presence of phages. Evolving phage resistance was costly, resulting in reduced carrying capacity, biofilm formation, and virulence in planta, possibly due to loss of QS-mediated expression of key virulence genes. We also observed mucoid phage-resistant colonies that showed loss of virulence and reduced twitching motility likely due to parallel mutations in prepilin peptidase gene, pilD. Moreover, phage-resistant SCVs from 35 °C-phage treatment had parallel mutations in type II secretion system (T2SS) genes (gspE and gspF). Adsorption assays confirmed the role of pilD as a phage receptor, while no loss of adsorption was found with phcS or T2SS mutants, indicative of other downstream phage resistance mechanisms. Additional transcriptomic analysis revealed upregulation of CBASS and type I restriction-modification phage defense systems in response to phage exposure, which coincided with reduced expression of motility and virulence-associated genes, including pilD and type II and III secretion systems. Together, these results suggest that while phage resistance-virulence trade-offs are not affected by the growth temperature, they could be mediated through both pre- and postinfection phage resistance mechanisms.

Detection of acrylamide in food based on MIL-glucose oxidase cascade colorimetric aptasensor.

BACKGROUND: Maillard reaction involves the polymerization, condensation, and other reactions between compounds containing free amino groups and reducing sugars or carbonyl compounds during heat processing. This process endows unique flavors and colors to food, while it can also produce numerous hazards. Acrylamide (AAm) is one of Maillard's hazards with neurotoxicity and carcinogenicity, these effects can trigger mutations and alternations in gene expression in human cells and accelerate organ aging. An accurate and reliable acrylamide detection method with high sensitivity and specificity for future regulatory activities is urgently needed. RESULTS: Herein, we constructed a colorimetric aptasensor with the hybridization of MIL-glucose oxidase (MGzyme)-cDNA and magnetic nanoparticle-aptamer (MNP-Apt) to specifically detect AAm. The incorporation of MB-Apt and AAm released MGzyme-cDNA in the supernatant, took the supernatant out, with the addition of glucose and TMB, MGzyme would oxidize glucose, the resulting •OH facilitated the oxidation of colorless TMB to blue ox-TMB. The absorbance value at 652 nm, which indicates the characteristic absorption peak of ox-TMB, exhibited a proportion to the concentration of AAm. MGzyme avoided the addition of harmful intermediate H2O2 and created an acid microenvironment for the catalytic reaction. MNP-Apt possessed the advantages of high specificity and simplified separation. Under optimal conditions, this method displayed a linear range of 0.01-100 µM with the limit of detection of 1.53 nM. With the spiked analysis data cross-verified by ELISA kit, this aptasensor was proven to specifically detect AAm at low concentrations. SIGNIFICANCE: This colorimetric aptasensor was the integration of aptamer and the enzyme-cascade system, which could broaden the applicable range of enzyme-cascade system, break the limits of specific detection of substrates, eliminate the need for harmful intermediates, improve the reaction efficiency, implement the specific detection, whilst enabling the accurate detection of AAm. Given these remarkable performances, this method has shown significant potential in the field of food safety inspection.

Effects of 8 months of high-intensity interval training on physical fitness and health-related quality of life in substance use disorder.

Objective: This study aimed to investigate the effect of 8 months of high-intensity interval training (HIIT) on physical fitness and health-related quality of life in substance use disorder. Methods: Sixty substance use disorder were randomly assigned to either the HIIT group or the control group according to a random sampling method. The HIIT group received 8 months of four 60-min sessions per week under supervision. Weight, waist circumference, body fat percentage, heart rate, blood pressure, VO2max, reaction time, grip strength, standing on one foot with eyes closed, sitting forward flexion, and quadrant jumping, standing on one foot with eyes closed, the number of push-ups, quality of life (SF-36) score, and craving (VAS) scored were monitored in the HIIT and control groups at baseline, 4 months, and 8 months. SPSS 22.0 was used to conduct repeated measurement analysis of variance and Pearson correlation analysis on the collected subject data. Results: Compared with baseline, weight (p < 0.001), waist circumference (p < 0.001), body fat percentage (p < 0.001), heart rate (p < 0.05), Systolic blood pressure (p < 0.01), systolic blood pressure (p < 0.05), reaction time (p < 0.001),PSQI (p < 0.001), Total cholesterol (p < 0.001), Triglyceride (p < 0.001), Blood sugar (p < 0.001) and VAS score (p < 0.001) were significantly decreased after 8 months of exercise intervention. Contrastingly, VO2max (p < 0.05), grip strength (p < 0.05), eyes closed and one foot Standing (p < 0.001), sitting forward flexion (p < 0.001), quadrant jumping (p < 0.001), push-ups (p < 0.001), PCS (p < 0.001), and MCS (p < 0.001) were significantly increased. VO2max was significantly negatively correlated with VAS (r = -0.434, p < 0.001), and significantly positively correlated with PCS (r = 0.425, p < 0.001). There was a positive correlation between standing on one foot with closed eyes and MCS (r = 0.283, p < 0.05). Conclusion: Eight months of HIIT can comprehensively improve the physical health level and health-related quality of life of men with substance use disorders, reduce the desire for drugs, and lay the foundation for better starting a happy life.

Effects of Adverse Events and 12-Week Group Step Aerobics on Sleep Quality in Chinese Adolescents.

BACKGROUND: In China, sleep disorders have become a public health concern. This study aimed to model the relationship between adverse events and sleep quality, as well as the effect of group step aerobics on sleep quality. METHODS: The modeling was built on surveying 2760 16-19-year-old adolescents. The Pittsburgh Sleep Quality Index (PSQI) was used to evaluate sleep quality, and the Adolescent Self-rating Life Events Checklist (ASLEC) was used to evaluate adverse events. Adolescents with sleep disorders (PSQI ≥ 8) were randomized into the control (n = 26) and exercise (n = 26) groups. The exercise group participated in 12-week step aerobics, and the 300 min weekly volume is compliant with the WHO physical activity guidelines. RESULTS: The double Poisson distribution was chosen to fit the data. ASLEC had a nonlinear relationship with the PSQI. Participants in the exercise group slept better (p < 0.05) from the eighth week until the end of the study. A random adolescent, therefore, has a 92.5% probability of experiencing improved sleep quality after 12 weeks of step aerobics. CONCLUSIONS: Intervention should be implemented before adverse events accumulate. An active lifestyle should be a preparedness strategy for increasing the resilience of adolescent mental health in the face of adversity.

Ratiometric SERS aptasensing for simultaneous quantitative detection of histamine and tyramine in fishes.

Herein, a SiO2@Ag NPs core/shell nanoparticles were synthesized to fabricate a surface-enhanced Raman spectroscopy (SERS) sensor for the simultaneous determination of histamine (HIS) and tyramine (TYR) based on specific aptamer recognition and ratiometric strategy. SiO2@Ag NPs with 4-thiosaminophenol (4-ATP) and Nile blue A (NBA) molecules were used as an internal standard (IS) and labeled with aptamers corresponding to HIS and TYR, respectively. Raman reporter molecules ROX and Cy5 labeled complementary DNA (cDNA) were then hybridized with aptamers to form rigid double-stranded DNA. After the HIS and TYR were captured by their aptamers, resulting in the dissociation of cDNA and separated from the SERS substrate. Therefore, the SERS signal intensity at 1503 cm-1 of ROX and 1358 cm-1 of Cy5 tagged on the terminal of cDNA decreased with the concentration of HIS and TYR increasing, while the SERS signal intensity at 1079 cm-1 of 4-APT and 592 cm-1 of NBA on the substrate remain stable. Thus, the concentrations of HIS and TYR can be determined by the I1503/I1079 and I1358/I592 values, respectively. This sensing strategy achieves a lower detection limit of 0.2 ng/mL for HIS and 0.05 ng/mL for TYR, respectively, demonstrating promising applications in sensitive detection of BAs in animal-derived foodstuff.

Prognostic influence of PD-1/PD-L1 suppressors in combination with chemotherapeutic agents for non-small cell pulmonary carcinoma: system review and meta-analysis.

Background: Lung cancer is a common malignant tumor, which is seriously harmful to human life and health. Nowadays, it has gradually become one of the best treatments for non-small cell lung cancer (NSCLC) to combine immunotherapy and chemotherapy, and its clinical efficacy is preliminary. Nevertheless, substantial differences exist between various studies and various indicators. Despite their unconvincing results, high-quality research evidence is needed to support them. In this case, further correlative studies are necessary to investigate the prognostic outcomes of PD-1/PD-L1 suppressors in combination with chemotherapeutic drugs in NSCLC. Methods: The online public databases were searchable for the clinical trials that consisted of NSCLC patients who had concluded their chemotherapy and who had accepted PD-1/PD-L1 suppressors. The time-span of the search spanned from the beginning to the end of the database. Two investigators retrieved the data independently. RevMan 5.3 statistical software was utilized for the assessment of bias risk. The software followed the Cochrane Handbook 5.3 guidelines. Results: There were seven clinically controlled studies with 2781 NSCLC samples finally included in this study. A meta-analysis of the post-treatment overall response rate (ORR) was undertaken. A remarkably higher ORR rate was observed in the study group (p<0.05). Study participants had a noticeably longer PFS (HR=0.61, 95% CI=0.54-0.70, P<0.00001). Study participants had markedly longer overall survival (OS) (HR=0.651, 95% CI=0.52-0.82, P<0.05). The incidence of adverse events (AEs) of Grade 3 or above was not clinically clearly different (P>0.05), as demonstrated by the incidence of AEs. The funnel plots were separately charted in accordance with ORR rate, PFE, OS, and Grade 3 AEs. The majority of the funnel plots were symmetrical and a minority of funnel plots were asymmetrical, indicating the heterogeneity of research and the limited evidence available may lead to some publication bias in the contained literature. Conclusion: The combined PD-1/PD-L1 inhibitors with conventional chemotherapy can dramatically elevate the prognosis of NSCLC patients, obviously enhancing the ORR rate and prolonging their PFS and OS. Furthermore, it was found that adding PD-1/PD-L1 inhibitors to conventional chemotherapy did not result in any additional adverse effects.

[Bamboo-based medicinal moxibustion for chronic fatigue syndrome: a randomized controlled trial].

OBJECTIVE: To observe the clinical efficacy of bamboo-based medicinal moxibustion for chronic fatigue syndrome (CFS), and to preliminarily explore its action mechanism. METHODS: Sixty-four patients with CFS were randomly divided into a moxibustion group (32 cases, 1 case dropped off, 1 case excluded) and an acupuncture group (32 cases, 2 cases dropped off). The patients in the moxibustion group were treated with bamboo-based medicinal moxibustion, while the patients in the acupuncture group were treated with routine acupuncture. Both groups were treated once a day, 6 days as a course of treatment with 1 day interval, for a total of 2 courses of treatment. Before treatment, 1 and 2 courses into treatment and in the follow-up of 14 days after treatment, the fatigue scale-14 (FS-14) and somatic and psychological health report (SPHERE) scores were observed in the two groups. Before and after treatment, the contents of CD+3, CD+4, CD+8 of peripheral blood T lymphocyte subsets were measured and CD+4/CD+8 ratio was calculated; the clinical efficacy of the two groups was compared. RESULTS: Compared before treatment, the FS-14 and SPHERE scores in the two groups were decreased 1 and 2 courses into treatment and in the follow-up (P<0.01), and the FS-14 and SPHERE scores in the moxibustion group were lower than those in the acupuncture group (P<0.01, P<0.05). Compared before treatment, the contents of CD+3, CD+4 and CD+4/CD+8 ratio in the moxibustion group were increased after treatment (P<0.01). There was no significant difference of CD+3, CD+4, CD+8 and CD+4/CD+8 ratio between before and after treatment in the acupuncture group (P>0.05). After treatment, the contents of CD+3 and CD+4 in the moxibustion group were higher than those in the acupuncture group (P<0.05). The total effective rate was 93.3% (28/30) in the moxibustion group, which was higher than 73.3% (22/30) in the acupuncture group (P<0.05). CONCLUSION: Bamboo-based medicinal moxibustion could improve the physical and mental fatigue symptoms and psychological status in patients with CFS. Its effect may be related to regulating the contents of CD+3, CD+4 of peripheral blood T lymphocyte subsets and CD+4/CD+8 ratio.

Rhizosphere phage communities drive soil suppressiveness to bacterial wilt disease.

BACKGROUND: Bacterial viruses, phages, play a key role in nutrient turnover and lysis of bacteria in terrestrial ecosystems. While phages are abundant in soils, their effects on plant pathogens and rhizosphere bacterial communities are poorly understood. Here, we used metagenomics and direct experiments to causally test if differences in rhizosphere phage communities could explain variation in soil suppressiveness and bacterial wilt plant disease outcomes by plant-pathogenic Ralstonia solanacearum bacterium. Specifically, we tested two hypotheses: (1) that healthy plants are associated with stronger top-down pathogen control by R. solanacearum-specific phages (i.e. 'primary phages') and (2) that 'secondary phages' that target pathogen-inhibiting bacteria play a stronger role in diseased plant rhizosphere microbiomes by indirectly 'helping' the pathogen. RESULTS: Using a repeated sampling of tomato rhizosphere soil in the field, we show that healthy plants are associated with distinct phage communities that contain relatively higher abundances of R. solanacearum-specific phages that exert strong top-down pathogen density control. Moreover, 'secondary phages' that targeted pathogen-inhibiting bacteria were more abundant in the diseased plant microbiomes. The roles of R. solanacearum-specific and 'secondary phages' were directly validated in separate greenhouse experiments where we causally show that phages can reduce soil suppressiveness, both directly and indirectly, via top-down control of pathogen densities and by alleviating interference competition between pathogen-inhibiting bacteria and the pathogen. CONCLUSIONS: Together, our findings demonstrate that soil suppressiveness, which is most often attributed to bacteria, could be driven by rhizosphere phage communities that regulate R. solanacearum densities and strength of interference competition with pathogen-suppressing bacteria. Rhizosphere phage communities are hence likely to be important in determining bacterial wilt disease outcomes and soil suppressiveness in agricultural fields. Video Abstract.

Development and validation of a label-free colorimetric aptasensor based on the HCR and hemin/G-quadruplex DNAzyme for the determination of patulin in fruits and fruit-based products from Xinjiang (China).

In this study, a simple, novel and practical label-free colorimetric aptasensor was successfully prepared for the ultrasensitive detection of patulin, based on the hybridization chain reaction (HCR) and hemin/G-quadruplex DNAzyme-signal amplification strategy. In this aptasensor, a detection probe was designed consisting of the aptamer sequence for the patulin and an initiator sequence to trigger the HCR. Two hairpin structures (H1 and H2) that included the G-quadruplex sequences in inactive configuration were used as functional elements. The presence of patulin triggered the opening of the hairpin structure and the beginning of the HCR. After the addition of hemin, G-rich DNA self-assembled into the peroxidase-mimicking hemin/G-quadruplex DNAzymes, which catalyzed a colorimetric reaction. Under optimized conditions, patulin was measured within a linear range of 0.1-200 ng mL-1, and the detection limit was 0.060 ng mL-1. The recovery rates ranged from 91.4 to 105% for fruits and fruit-based products. Subsequently, a total of 311 samples comprising fruits, fruit-based products and dried fruits were collected from supermarkets, production bases and farmers' markets in Xinjiang, and analyzed for patulin using the proposed aptasensor. Patulin was detected in 16 samples (5.14%) at concentrations ranging from 1.23 to 16.4 µg kg-1. None of the samples exceeded the maximal level set by the EU commission (50 µg kg-1). The positivity in fresh fruits (7.69%) was significantly higher than that of fruit-based products (4.00%) and dried fruits (1.25%). In summary, the proposed aptasensor can quickly detect patulin in food samples, thus providing a warning for mycotoxin contamination.

Selecting specific aptamers that bind to ovine pregnancy-associated glycoprotein 7 using real serum sample-assisted FluMag-SELEX to develop magnetic microparticle-based colorimetric aptasensor.

The pregnancy-associated glycoproteins (PAGs) have been widely used as biomarkers for the early diagnosis of pregnancy in cattle and sheep. This study aimed to obtain the single-stranded DNA aptamers that specifically bound to ovine pregnancy-associated glycoprotein 7 (ovPAG7) with high affinity (Kd = 9.8-32.4 nM) using real serum sample-assisted FluMag-systematic evolution of ligands by exponential enrichment (SELEX). Subsequently, the selected aptamers were applied to fabricate an ultrasensitive colorimetric aptasensor for ovPAG7 detection based on functionalized magnetic microparticles and hybridization chain reaction. Under the optimized conditions, the colorimetric aptasensor exhibited a broad linear range (0.2-500 ng mL-1), low detection limit (0.081 ng mL-1), good recovery rate (94.5-109.1%), and high repeatability (relative standard deviation of 4.02-8.16%) in ovPAG7-spiked serum. Furthermore, this aptasensor was applied to measure the ovPAG7 in serum samples of ewes for pregnancy diagnosis. Blood samples were collected from Chinese Merino ewes on days 22, 28 after artificial insemination (AI) for ovPAG7 detection, respectively. Transrectal ultrasonography diagnosis of pregnancy 45 days after AI was the reference (gold) standard for all PAG tests. Diagnostic sensitivity, specificity, and accuracy of the proposed aptasensor were 95.8, 87.5, and 92.5% at day 22 and 95.8, 90.6, and 93.7% at day 28, respectively. The degree of agreement (Kappa) between developed aptasensor and ultrasonography diagnosis 22 and 28 day after AI were higher than 0.8. These results illustrated that the aptasensor was proved to be a sensitive, reliable and cost-effective way of measuring PAG and might be a useful means of pregnancy detection in ewes.

Immunocompetent rabbits infected with Cryptosporidium cuniculus as an animal model for anti-cryptosporidial drug testing.

Cryptosporidium is one of the leading causes of diarrheal disease in humans and animals, which can be severe and deadly in neonates and immunocompromised hosts. Studies on the biology of Cryptosporidium and drug discovery efforts have been hindered by a number of factors including the limited availability of animal models. Here, we report the establishment and characterization of an immunocompetent rabbit model for infection with Cryptosporidium cuniculus. By testing four known anti-cryptosporidial compounds (nitazoxanide, baicalein, curcumin and matrine), we showed that the rabbit could be used as an alternative animal model for evaluating anti-cryptosporidial drug efficacy in vivo.

Identification and validation of key long non-coding RNAs using co-expression network analysis in Crohn's disease.

BACKGROUND: Crohn's disease (CD) is a chronic inflammatory disease of the digestive tract. The underlying molecular mechanism of CD remains unclear. The aim of this study was to investigate the differentially expressed long non-coding RNA (lncRNA) in CD and its possible mechanism, and to verify the expression of lncRNA. METHODS: Microarray GSE67106 and GSE83448 were downloaded from the Gene Expression Omnibus (GEO) database. Differentially expressed lncRNAs (DELs) and messenger RNAs (mRNAs, DEGs), when normalized through the betaqn package in the R, were determined via the limma package. Gene Ontology (GO) and Kyoto Encyclopedia of genes and genomes (KEGG) pathways were studied using the database for the annotation, visualization and integrated discovery (DAVID) version 6.7, along with Gene Set Enrichment Analysis (GSEA) version 3.0. The co-expression of lncRNAs-mRNAs were determined using weighted gene co expression network analysis (WGCNA). The micro RNAs (miRNAs) related to the DELs and DEGs were forecast. A competing endogenous RNA (ceRNA) network was established. RESULTS: There were 42 DEGs and 551 DEGs identified in total among the samples of the CD and normal control, respectively. These DEGs were enriched in such pathways as retinol metabolism, renin angiotensin system, and maturation-related signaling pathways. A lncRNA-mRNA co-expression network was constructed by WGCNA, with CDKN2B-AS (ANRIL), CTC-210G5.1.1, RP11-467L20.10.1, RP11-325F22.5.1, and RP11-59E19.1.1 as hub DELs. Together with miRNAs, a ceRNA network was constructed and functional analysis showed that the cell brush border and plasma membrane, synthesis and transport of lipoprotein, and angiotensin maturation, metabolism, and regulation of blood pressure were involved in the progression of CD. We successfully validated 1 lncRNA ANRIL, in our clinical specimens, ANRIL, which can feature prominently in CD. However, the exact mechanism of lncRNA ANRIL in CD prediction and diagnosis requires further exploration. CONCLUSIONS: This study showed that lncRNA ANRIL has a certain predictive effect on CD occurrence and development and could be a new potential treatment target.

[Different stimulation methods on auricular points for primary dysmenorrhea: a randomized controlled trial].

OBJECTIVE: To prove the therapeutic effect of auricular intradermal needling and auricular point sticking on primary dysmenorrhea (PD), and to explore its mechanism. METHODS: A total of 90 patients with PD were randomized into an auricular intradermal needling group, an auricular point sticking group and a placebo group, 30 cases in each one. Neishengzhiqi (TF2), Neifenmi (CO18), Shenmen (TF4), Shen (CO10), Jiaogan (AH6a), Gan (CO12) and Pizhixia (AT4) were selected in the 3 groups, intradermal needling and cowherb seed sticking were applied respectively in the auricular intradermal needling group and the auricular point sticking group, adhesive tape without needle was stuck in the placebo group. Pressing and kneading for 3 to 4 times were required each day, 3 to 4 min each time, and the intervention was started 5 d before menstruation, once every other day, 4 times each menstrual cycle were as one course, and totally 3 courses were required in the 3 groups. The follow-up was adopted at the next menstruation after treatment. The Cox menstrual symptom scale (CMSS) score, the visual analogue scale (VAS) score and the self-rating anxiety scale (SAS) score before treatment, 1,2,3 courses into treatment and at follow-up were compared, the serum levels of PGF2αand PGE2 before and after treatment were detected, and the clinical therapeutic effect was evaluated in the 3 groups. RESULTS: Compared before treatment, the scores of CMSS, VAS and SAS were decreased at each time point of treatment in the auricular intradermal needling group, 2, 3 courses into treatment and at follow-up in the auricular point sticking group and 3 courses into treatment in the placebo group (P<0.001, P<0.05). Compared with the auricular point sticking group, the CMSS scores at each time point of treatment and the VAS scores of 1, 2 courses and at follow-up were decreased in the auricular intradermal needling group (P<0.05). Compared with the placebo group, the CMSS scores were decreased at each time point of treatment in the auricular intradermal needling group and 3 courses into treatment and at follow-up in the auricular point sticking group (P<0.001, P<0.05); the VAS scores were decreased at each time point of treatment in the auricular intradermal needling group and the auricular point sticking group (P<0.001, P<0.05). After treatment, the serum levels of PGF2α were decreased (P<0.05) and the serum levels of PGE2 were increased (P<0.05) in the auricular intradermal needling group and the auricular point sticking group, and the serum levels of PGF2α were lower than the placebo group (P<0.05), the serum levels of PGE2 were higher than the placebo group (P<0.05) in the two groups. Compared with the auricular point sticking group, the serum level of PGF2 was decreased (P<0.05), the serum level of PGE2α was increased in the auricular intradermal needling group (P<0.05). The total effective rates were 93.3% (28/30) in the auricular intradermal needling group and 80.0% (24/30) in the auricular point sticking group, which were both superior to 63.3% (19/30) in the placebo group (P<0.05). CONCLUSION: Auricular intradermal needling and auricular point sticking can both improve the clinical symptom of primary dysmenorrhea, relieve the pain and anxiety, their mechanism may be related to regulating the serum levels of PGF2α and PGE2. The therapeutic effect of auricular intradermal needling is superior to auricular point sticking, and the placebo effect can be preliminarily excluded.

Bioinformatics analysis, codon optimization and expression of ovine pregnancy associated Glycoprotein-7 in HEK293 cells.

Pregnancy-associated glycoproteins (PAGs) are widely used as powerful markers for early pregnancy diagnosis in livestock. To improve expression efficiency of recombinant ovine pregnancy-associated glycoprotein-7 (ovPAG-7) in HEK293 cells, the codon usage bias of the ovPAG-7 gene was analyzed using bioinformatic approaches, after which the DNA sequence encoding ovPAG-7 was designed, synthesized, and expressed in HEK293. The structure and function of ovPAG-7 were predicted using bioinformatics software and online databases. The results showed that the effective number of codons (NEC) of the ovPAG-7 gene was 56.82, indicating that the ovPAG-7 gene was weakly biased. ovPAG-7 gene had 26 biased codons (relative synonymous codon usage (RSCU) > 1), 15 of which were biased towards G/C at the third position. After codon optimization, the codon adaptation index of the ovPAG-7 gene increased from 0.74 to 0.96, and its GC content changed from 46.6 to 58.6%. The amino acid sequence encoded by the optimized gene was entirely consistent with those published in Gen Bank. Western blot analysis indicated that the recombinant ovPAG-7 protein with a relative molecular mass of 48 kDa was successfully expressed in HEK293 cells. The bioinformatics prediction results showed that ovPAG-7 protein contained 3 N-glycosylation sites, 13 B-cell epitopes, and a signal peptide consisting of 15 amino acids at the N terminus. The secondary structure of the ovPAG-7 protein was predicted to consist of random coils (46.85%), extended strands (32.05%), α-helices (16.16%), and ß-turns (4.93%). This study provided a tool for the screening of monoclonal antibodies and functional research on ovPAG-7.

Paraventricular Calcitonin Receptor-Expressing Neurons Modulate Energy Homeostasis in Male Mice.

The paraventricular nucleus of the hypothalamus (PVH) is a heterogeneous collection of neurons that play important roles in modulating feeding and energy expenditure. Abnormal development or ablation of the PVH results in hyperphagic obesity and defects in energy expenditure whereas selective activation of defined PVH neuronal populations can suppress feeding and may promote energy expenditure. Here, we characterize the contribution of calcitonin receptor-expressing PVH neurons (CalcRPVH) to energy balance control. We used Cre-dependent viral tools delivered stereotaxically to the PVH of CalcR2Acre mice to activate, silence, and trace CalcRPVH neurons and determine their contribution to body weight regulation. Immunohistochemistry of fluorescently-labeled CalcRPVH neurons demonstrates that CalcRPVH neurons are largely distinct from several PVH neuronal populations involved in energy homeostasis; these neurons project to regions of the hindbrain that are implicated in energy balance control, including the nucleus of the solitary tract and the parabrachial nucleus. Acute activation of CalcRPVH neurons suppresses feeding without appreciably augmenting energy expenditure, whereas their silencing leads to obesity that may be due in part due to loss of PVH melanocortin-4 receptor signaling. These data show that CalcRPVH neurons are an essential component of energy balance neurocircuitry and their function is important for body weight maintenance. A thorough understanding of the mechanisms by which CalcRPVH neurons modulate energy balance might identify novel therapeutic targets for the treatment and prevention of obesity.

Long-term efficacy and safety of monotherapy with a single fresh fecal microbiota transplant for recurrent active ulcerative colitis: a prospective randomized pilot study.

BACKGROUND: To assess the long-term safety and efficacy of monotherapy with a single fresh fecal microbiota transplant (FMT) for recurrent ulcerative colitis (UC). RESULTS: Twenty-six eligible patients were enrolled, and 6 patients were excluded. Ultimately, 20 patients were randomized to the FMT group (n = 10) and the control group (n = 10); 80% were females (F/M = 16/4), the mean age was 48 ± 14 years, and the mean duration was 6.4 ± 8.2 years. The mean length of post-FMT follow-up was 19.1 ± 10.1 months (6-38). No statistically significant differences in baseline demographic or clinical characteristics were found between the groups. Ninety percent of patients in the FMT group and 50% of patients in the control group met the primary endpoint at week 8. The Mayo score was significantly decreased compared with that of the control group (n = 10) when reassessed at week 4 (P = 0.001) and week 8 (P = 0.019) after FMT; there was no significant difference 6 months after treatment. The median remission time was 24 months (95% CI 68.26-131.7%) in both the FMT (range 6-38 months) and control groups (range 7-35 months), with no significant difference (P = 0.895). Participants tolerated FMT treatment, and no adverse events occurred during long-term follow-up, with one treatment-related significant adverse event (EBV infection) occurring within 2 weeks after FMT. Stool microbiota composition analysis indicated improved gut microbiota diversity after FMT, with expansion of stool-donor taxa. Bacteroidetes, Firmicutes and Proteobacteria were the dominant bacterial phyla of the gut microbiota in active UC patients. The relative abundance of Bacteroidetes decreased and that of Proteobacteria increased significantly in active UC patients compared with donors, while Firmicutes showed no significant changes. A single fresh FMT could effectively reconstruct the gut microbiota composition in patients with active UC and maintain stability, with increased Bacteroidetes and decreased Proteobacteria abundance. FMT significantly reduced the relative abundance of Escherichia and increased the relative abundance of Prevotella at the genus level. Pyruvate metabolism, glyoxylate and dicarboxylate metabolism, and pantothenate and CoA biosynthesis showed significant differences after transplantation. CONCLUSIONS: Monotherapy with a single fresh FMT is an effective and safe strategy to induce long-term remission without drugs in patients with active UC and may be an alternative induction therapy for recurrent UC or even primary UC.

Peripheral Inflammatory Biomarkers of Methamphetamine Withdrawal Patients Based on the Neuro-Inflammation Hypothesis: The Possible Improvement Effect of Exercise.

Methamphetamine (MA) induced addiction and neuroinflammation has been implicated. Based on the neuroinflammation hypothesis, this study aims to investigate how exercise influences the craving of patients in MA withdrawal, and explore the mechanism of peripheral inflammation. A total of 90 patients in MA withdrawal were recruited. No difference was noted in the number of years of drug use and the frequency of drug use among patients, and the withdrawal time was within 2 months. The subjects were grouped based on the degree of craving induced by the cues: non-craving control group (NCC group), craving control group (CC group), and craving exercise group (CE group). The CE group was subjected to aerobic combined resistance training. Then, the ELISA method was used to detect plasma IL-6, TNF-α, and IL-1ß concentrations; Visual Analog Scale (VAS) measurement of cue-induced cravings under Virtual Reality (VR) exposure (VR-VAS) and the Desires for Drug Questionnaire (DDQ) were used to assess cravings. Consequently, plasma IL-6, TNF-α, IL-1ß, levels, and the VR-VAS and DDQ scores of MA withdrawal patients were significantly reduced after exercise. This study confirmed that 8 weeks of incremental load aerobic combined with resistance training reduces peripheral inflammation and significantly reduces the level of craving for MA.

Selection, identification, and application of DNA aptamers against bovine pregnancy-associated glycoproteins 4.

The bovine pregnancy-associated glycoproteins (bPAGs) have been widely used as robust markers for early diagnosis of pregnancy in the cattle. The current immune recognition methods for detecting bPAGs are limited and, to a certain extent, are associated with high costs and poor stability of the antibody. Aptamers that are more stable and easily synthesized than antibodies might serve as suitable candidates for the development of rapid detection methods. This paper describes selection and characterization of bPAG4 aptamers and theirs applicability to detect bPAG4 in the serum. In this work, the recombinant bovine pregnancy-associated glycoproteins 4 (bPAG4) with a relative molecular mass of about 48 kDa was successfully expressed in human embryonic kidney 293 (HEK 293) cells. Subsequently, the ssDNA aptamers were selected by systematic evolution of ligands by exponential enrichment (SELEX) using magnetic beads (MB) coated with bPAG4 as target. After 9 rounds of selection, three aptamers with high affinity to bPAG4 (Kd = 11.7~40.2 nM) were identified. The selected aptamers were successfully used in enzyme-linked aptamer assay (ELAA) to detect bPAG4 at a detection limit of 0.09 ng/mL. Meanwhile, it has been successfully applied for the detection of bPAG4 in serum samples. This work demonstrated that the selected aptamers could be used as promising affinity probes in the development of inexpensive, simple, and sensitive analysis methods for detecting bPAGs. Graphical abstract.