The aim of this investigation was to determine the optimal storage medium for testicular hypothermic transportation and identify the ideal concentration for the application of the protective agent 5-aminolevulinic acid (5-ALA). Furthermore, this study aimed to explore the underlying mechanism of the protective effects of 5-ALA. First, we collected and stored mouse testicular fragments in different media, including Hank's balanced salt solution (HBSS; n = 5), Dulbecco's Modified Eagle Medium/Nutrient Mixture F-12 (DMEM/F12; n = 5), and alpha-minimum essential medium (αMEM; n = 5). Storage of testicular tissue in HBSS preserved the integrity of testicular morphology better than that in the DMEM/F12 group ( P < 0.05) and the αMEM group ( P < 0.01). Testicular fragments were subsequently placed in HBSS with various concentrations of 5-ALA (0 [control], 1 mmol l -1 , 2 mmol l -1 , and 5 mmol l -1 ) to determine the most effective concentration of 5-ALA. The 2 mmol l -1 5-ALA group ( n = 3) presented the highest positive rate of spermatogonial stem cells compared with those in the control, 1 mmol l -1 , and 5 mmol l -1 5-ALA groups. Finally, the tissue fragments were preserved in HBSS with control ( n = 3) and 2 mmol l -1 5-ALA ( n = 3) under low-temperature conditions. A comparative analysis was performed against fresh testes ( n = 3) to elucidate the underlying mechanism of 5-ALA. Gene set enrichment analysis (GSEA) for WikiPathways revealed that the p38 mitogen-activated protein kinase (MAPK) signaling pathway was downregulated in the 2 mmol l -1 5-ALA group compared with that in the control group (normalized enrichment score [NES] = -1.57, false discovery rate [FDR] = 0.229, and P = 0.019). In conclusion, these data suggest that using 2 mmol l -1 5-ALA in HBSS effectively protected the viability of spermatogonial stem cells upon hypothermic transportation.
Male ; Animals ; Testis/cytology* ; Aminolevulinic Acid/pharmacology* ; Mice ; Organ Preservation/methods* ; Organ Preservation Solutions/pharmacology* ; Cryopreservation/methods*

Direct electrical stimulation of the human cortex can produce subjective visual sensations, yet these sensations are unstable. The underlying mechanisms may stem from differences in electrophysiological activity within the distributed network outside the stimulated site. To address this problem, we recruited 69 patients who experienced visual sensations during invasive electrical stimulation while intracranial electroencephalography (iEEG) data were recorded. We found significantly flattened power spectral slopes in distributed regions involving different brain networks and decreased integrated information during elicited visual sensations compared with the non-sensation condition. Further analysis based on minimum information partitions revealed that the reconfigured network interactions primarily involved the inferior frontal cortex, posterior superior temporal sulcus, and temporoparietal junction. The flattened power spectral slope in the inferior frontal gyrus was also correlated with integrated information. Taken together, this study indicates that the altered electrophysiological signatures provide insights into the neural mechanisms underlying subjective visual sensations.
Humans ; Male ; Female ; Adult ; Visual Perception/physiology* ; Electric Stimulation ; Middle Aged ; Young Adult ; Electrocorticography ; Electroencephalography ; Brain Mapping

Image 1.

The biological clock(also known as the circadian rhythm)is the fundamental reliance for all organisms on Earth to adapt and survive in the Earth's rotation environment.Circadian rhythm is the most basic regulatory mechanism of life activities,and plays a key role in maintaining normal physiological and biochemical homeostasis,disease occurrence and treatment.Recent studies have shown that the biologi-cal clock plays an important role in the development of oral tissues and in the occurrence and treatment of oral diseases.Since there is cur-rently no guiding literature on the research methods of biological clock in stomatology,researchers mainly conduct research based on pub-lished references,which has led to controversy about the research methods of biological clock in stomatology,and there are many confusions about how to rationally apply the research methods of circadia rhythms.In view of this,this expert consensus summarizes the characteristics of the biological clock and analyzes the shortcomings of the current biological clock research in stomatology,and organizes relevant experts to summarize and recommend 10 principles as a reference for the rational implementation of the biological clock in stomatology research.

Objective To investigate the role of member septin family(SEPT12)in human spermatogenesis and its influence on sperm motility and sperm ultrastructure.Methods Whole exome sequencing(WES)was performed on peripheral blood DNA extracted from 375 patients with asthenoteratozoospermia,and a patient with idiopathic in-fertility carrying compound heterozygous mutation of SEPT12 was screened out.Sanger sequencing was performed to verify the mutation,and co-segregation analysis was performed in the family.The morphological abnormalities of sperm were analyzed by hematoxylin-eosin(HE)staining and scanning electron microscopy(SEM),and the ultra-structural defects of sperm were analyzed by transmission electron microscopy(TEM).Then the effects of the muta-tion on the level and position of the protein and the changes of the location and level of the defect structure markers were analyzed by Western blot and immune-fluorescence(IF).Results The compound heterozygous mutations c.C332A(p.Ti111K)and c.406_416 del TGCTCGTATTG(p.q136 VFS*39)in the SEPT12 gene were screened and identified in a patient with asthenoteratozoospermia.The mutations were verified by Sanger sequencing,which was consistent with the co-segregation genetic pattern of the family.The mutations resulted in loss of protein expres-sion,decreased sperm motility and sperm morphological deformities,mainly including short tail,curly tail and ir-regular sperm head.The ultrastructure of sperm showed that the annulus between the mid-piece and the principle-piece was missing,the acrosome membrane of sperm head fell off and the nucleus contained vacuoles.In the mid-piece of sperm flagella,the arrangement of mitochondrial sheath was disordered,most of flagella axoneme central pair was absent,microtubules doublet was missing or disordered,and some radical spoke was absent.By Western blot and IF,the marker proteins of related structural components were detected,and the results showed that the level of SEPT4 protein decreased,SEPT6 protein unchanged,acrosomal related proteins ACTL7A and ACROSIN protein missing,and the expression levels of mitochondrial and axoneme related proteins TOMM20,SPAG6 and RSPH3 protein significantly decreased.Conclusion The deletion of SEPT12 protein caused by SEPT12 gene mu-tation leads to the deletion of the annulus between the mid-piece and the principle-piece,and the abnormal assem-bly of sperm acrosome,mitochondrial sheath and flagella.

Objective To establish a simultaneous detection approach for 34 emerging contaminants(ECs)in tap water by liquid chromatography-tandem mass spectrometry(HPLC-MS/MS).Human health risk assessment was performed according to the detection results from 43 tap water samples.Methods Tap water samples were concentrated and extracted by solid phase extraction,and then blown to near dry by nitrogen at 40℃.The sample extracts were dissolved in methanol-water solution(95:5,VN)to 0.5 mL for analyzing.Agilent Jet Stream Electrospray Ionization(AJS ESI)and the multiple reaction monitoring(MRM)mode were performed for MS to acquire the data of 34 ECs.A database including precursor ion,product ion and retention times was established accordingly.Results The average linear correlation coefficients(r)of 34 kinds of ECs was 0.995 9.The limits of detection were 0.01～0.60 ng/L and the recoveries were between 60.7％and 119.8％.The intra-group precisions were between 0.05％～9.89％and the intra-day precisions were between 0.20％～14.40％for the spiked samples.The method was applied to analyze 43 tap water samples and a total of 15 ECs were detected.According to the results,the detection rate of caffeine was the highest(84％),and the concentration range was ND～74.42 ng/L.Among all the ECs detected,1,2,3-benzotriazole had the highest concentration(ND～361.15 ng/L),where detection rate was 44％.Humans may be exposed to these ECs by drinking the tap water.The human health risk assessments of 12 kinds of ECs were carried out,however,the estimated risk was negligible(risk quotient＜0.01).Conclusion The method is simple,highly sensitive and selective,and could meet the detection needs of ECs at trace level in tap water.There was no human health risk posed for ECs identified in 43 tap water samples analyzed by this method.

Objective To investigate the correlation between bone mineral density(BMD)at different hip positions with muscle parameters and physical performance in middle-aged and elderly people in Kunming area.Methods 531 middle-aged and elderly volunteers were recruited from the Radiology Department of the First People's Hospital of Yunnan Province from May 2021 to April 2022.All study subjects completed the five-times-sit-to-stand test(FTSST)and hip quantitative CT(QCT)examinations.Volunteers'total hip(TH),femoral neck(FN),and intertrochanteric(IT)BMD were measured by using QCT PRO workstation and using OsiriX software to measure the area and density of their gluteus maximus muscle,gluteus medius,and minimus muscle and midthigh muscle.Divide male and female volunteers into positive and negative groups respectively based on FTSST time≥12 seconds or<12 seconds,and analyze the differences in hip BMD between the groups;Also divide male and female volunteers into three groups(50～59 years old,60～69 years old,70 years old and above)at the age of 10,and analyze the correlation between hip BMD and muscle parameters in different age and gender stratification.Control for age and BMI,and then perform partial correlation analysis on the above indicators.Results The BMD of the hip in the female FTSST positive group was lower than that in the negative group(P<0.001),while there was no statistically significant difference between the male groups(P>0.05).After adjusting for age and BMI,among males,FN BMD was positively correlated with gluteus medius and minimus muscle density in the age groups of 50～59 and 60～69(P<0.05),while TH BMD,FN BMD,and IT BMD were strongly positively correlated with gluteus medius and minimus muscle density in the age group over 70(P<0.05).For females,the correlation between hip BMD and muscle density in the age groups of 50～59 and 60～69 was weak,while BMD in all parts of the hip was not correlated with muscle density in the age group over 70(P>0.05).There was a negative correlation between TH BMD,FN BMD,and gluteus medius and minimus muscle area in the age group of 50～59 years old for males(P<0.05),while there was a significant negative correlation between TH BMD,IT BMD,and gluteus maximus area in the age group of 70 years old and above for females(P<0.05).Conclusion The BMD of various parts of the hip in the female FTSST positive group is lower than that in the negative group.The density of the gluteus medius and minimus muscle can to some extent serve as a predictive indicator of femoral neck bone strength in middle-aged and elderly men in Kunming area.

Objective : To investigate the drug resistance and pathogenicity of six clinical isolates of Klebsiella pneu- moniae (Kp) ，and to provide a basis for prevention and treatment of Kp infection． Methods : The six strains from different hospitals were isolated ，cultured ，and identified by species-specific gene khe． Their whole genome se- quences (WGS) were obtained using next-generation sequencing technology (NGS) ．Based on the WGS，the cap- sular serotypes，sequence types (ST) and drug-resistance genes of six strains were identified．The capsular sero- type genes and virulence genes were validated or identified using PCR. Broth microdilution tests were conducted to validate their drug susceptibility，and mice were challenged with Kp aerosols by MicroSprayer aerosolizer to evaluate their pathogenicity. Results : The six strains were all serotype K2 but belonged to four ST types ( ST14 ，ST65， ST700，and ST86) ，and collectively carried six virulence genes and 23 drug-resistance genes．All the six strains were resistant to ampicillin，but only one strain was multidrug-resistant．Four strains exhibited high mucoid charac- teristics．Five strains could cause mortality in mice，which were preliminary identified as high virulence strains. Conclusion For the six Kp clinical isolates from different sources，only one strain named NY 13294 is both multi- drug-resistant and highly virulent，and other four highly virulent strains are resistant to one or two types of antibiot- ics.