OBJECTIVES: To investigate the effects of early life intervention with Bifidobacterium bifidum BD-1 (B. bifidum BD-1) on hyperactivity in a female mouse model of attention deficit hyperactivity disorder (ADHD) and explore the underlying mechanisms. METHODS: Eight newborn female Wistar-Kyoto (WKY) rats and 6 spontaneous hypertensive rats (SHRs) were gavaged with saline and another 6 SHRs were gavaged with B. bifidum BD-1 (10⁹ CFU) daily for 3 weeks. Open field test of the rats was conducted at 7 weeks, and fecal samples were collected at weaning (3 weeks) and at 7 weeks for 16S rRNA sequencing. Immunofluorescent staining was used to detect dopamine transporter (DAT) and tyrosine hydroxylase (Th) levels in the striatum and activated microglia in the prefrontal cortex. Treg cells in the mesenteric lymph nodes, spleen and blood were analyzed using flow cytometry. RESULTS: The SHRs traveled a significantly greater distance in open fields test than WKY rats, and this behavior was significantly attenuated by B. bifidum BD-1 intervention. The expression of DAT and Th in the striatum was significantly lower in the SHRs than in WKY rats, while B. bifidum BD-1 treatment obviously increased Th levels in the SHRs. B. bifidum BD-1 intervention significantly deceased the number of activated microglia and increased Treg cell counts in the spleen of SHRs. The treatment also enhanced α diversity in gut microbiota of the SHRs and resulted in a decreased Firmicutes/Bacteroidota ratio, more active Muribaculaceae growth, and suppression of Clostridia_UCG-014 proliferation. CONCLUSIONS Early life intervention with B. bifidum BD-1 alleviates hyperactivity in female SHRs by modulating the gut microbiota and peripheral immune response, suppressing neuroinflammation and improving dopaminergic system function. These findings provide evidence for early prevention strategies and support the development and application of psychobiotics for ADHD.
Animals ; Female ; Rats ; Rats, Inbred WKY ; Rats, Inbred SHR ; Attention Deficit Disorder with Hyperactivity/therapy* ; Bifidobacterium bifidum ; Probiotics/therapeutic use* ; Dopamine Plasma Membrane Transport Proteins/metabolism* ; Tyrosine 3-Monooxygenase/metabolism* ; Gastrointestinal Microbiome ; Disease Models, Animal

Objective To make a preliminary investigation of the effect of the immune pathway mediated by live Lacticaseibacillus paracasei N11 15 on the development of primary hippocampal neurons cultured in vitro.Methods Live Lacticaseibacillus paracasei N1115 suspension of an appropriate concentration was used as the experimental group.Peptidoglycan(PGN)and lipopolysaccharide(LPS)were used as positive controls,and RPMI1640 medium served as the blank control.These were co-cultured with RAW264.7 cells to obtain the co-culture mediums and the total cellular RNA,and to measure the expression and secretion of cytokines.After centrifugation,the supernatants were co-cultured with primary hippocampal neurons at appropriate ratios.The co-culture mediums were collected,and the total cellular RNA was extracted to measure the expression of genes related to synaptic development in neurons.Following immunofluorescence staining of the primary hippocampal neurons,the presynaptic and presynaptic membrane-associated proteins,including synaptophysin(SYP)and the postsynaptic density protein 95(PSD95),and neuronal cell maturation markers,including microtubule-associated protein 2(MAP-2),and doublecortin(DCX)were quantitatively analyzed.Additionally,the morphological development of the neurons were measured.Results Compared with the blank control,the mRNA expression levels of interleukin(IL)-6 and tumor necrosis factor-α(TNF-α)increased by 7471％and 926％,respectively,after the RAW264.7 cells were treated with live Lacticaseibacillus paracasei N1115,while their secretion levels increased by 184.16 pg/mL and 12320.76 pg/mL,respectively,all showing statistically significant differences(P＜0.05).The activation ability of N1115 live bacteria was stronger than that of PGN but weaker than that of LPS,showing statistically significant differences(P＜0.05).Compared with the blank control,following the intervention with the supernatant from the co-culture of N1115 live bacteria and RAW264.7 cells,the viability of primary hippocampal neurons in the 10％supernatant intervention group increased by 19.25％,showing statistically significant differences(P＜0.05),and the mRNA expression of SYP and PSD95 increased by 137％and 159％,respectively,showing statistically significant differences(P＜0.05).The total neurite length(489.88 μm)of the neurons of the group intervened with the supernatant of N11 15 live bacteria was increased compared to that of the blank control group(381.51 μm),and the cell body area(2092.22 μm2),maximum neurite length(184.78 μm),total neurite length,average neurite length(108.38 μm),and branching points(4.84 s)were higher than those in the two positive control groups,showing statistically significant differences(P＜0.05).Conclusion Lacticaseibacillus paracasei N1115 can significantly activate the immune regulatory function of macrophages,thereby promoting the morphological development and synaptic function of nerve cells.

Objective To investigate the clinical value of real-time shear wave elastography( SWE) in early diagnosis of diabetic nephropathy ( DN ) . Methods One hundred and fifty-five patients with type 2 diabetes mellitus (DM) in our hospital were selected as DN group and 54 healthy volunteers with matched age-sex were selected as control group . According to the criteria of Mogensen staging ,patients of DN group were divided into 3 subgroups according to urine protein/creatinine ( ACR) :group A ( DM or DN Ⅰ ,Ⅱ ) , ACR＜30 mg/g ;group B ( DN Ⅲ ) ,ACR 30 －300 mg/g ;group C ( DN Ⅳ ,Ⅴ ) ,ACR＞ 300 mg/g . All subjects were examined by SWE technique to detect the maximum ,mean and minimum Young′s modulus values ( Emax ,Emean ,Emin) of the middle of left renal parenchyma ,the values of Young′s modulus in the control group and DN patients were compared . The relationship between Young′s modulus and ACR in patients with DN was analyzed . Results The Emax ,Emean and Emin in DN patients were all greater than those in control group ( P ＜0 .05) . In group A ,B and C ,Emax ,Emean and Emin gradually increased ( P＜0 .05) . Spearman correlation analysis showed that there was a positive correlation between Young′s modulus and ACR in patients with DN ( r ＝ 0 .817 ,0 .764 ,0 .609 ;all P＝ 0 .000) . ROC curve analysis showed that the area under the curve of Emax diagnosis DN Ⅲ stage was 0 .810 ,the cut-point value was 6 .65 kPa ,the sensitivity was 79 .0% ,and the specificity was 81 .7% ;the area under the curve of Emean diagnosis DN Ⅲ stage was 0 .785 ,and the cut-off value was 3 .69 kPa ,sensitivity 62 .8% ,specificity 78 .3% . Conclusions SWE technique can quantitatively analyze renal parenchyma hardness in patients with different stages of DN ,which may provide new diagnostic information for early diagnosis of DN .

Objective: To investigate the correlation between the maximum elastic modulus(Emax) of breast cancer obtained by shear-wave elastography (SWE) and immunohistochemical profiles. Methods: One hundred and sixty patients with 171 pathologically proven breast lesions were included in this study. All of them underwent conventional ultrasound and SWE examination before surgery. The Emax and maximum diameter were obtained. Histologic grade, immunohistochemical profiles ER, PR, HER-2, Ki-67 and molecular subtypes were recorded according to pathological results after surgery. The correlation between Emax and tumor size, histological grade, ER, PR, HER-2, Ki-67 and molecular subtypes were analyzed. Results: ①There was a positive correlation between the tumor size with Emax(r=0.510, P<0.05); ②The Emax of tumors with HER-2 positive expression were higher than that with HER-2 negative expression(P<0.05), the Emax of tumors with Ki67 high expression were higher than that with Ki67 low expression (P<0.05); ③There were significant differences in Emax between different molecular subtypes(P<0.05), the Emax of HER-2 positive tumor was the highest, followed by those of Luminal B tumor, triple-negative and Luminal A tumor; ④Multiple linear regression indicated that tumor size was the strongest pathologic determinate of Emax value(P<0.05), its standardized partial regression coefficient was 0.500, and HER-2 also had significant influence on Emax value of tumor(P<0.05), the standardized partial regression coefficient was 0.237. Conclusions Some prognosis immunohistochemical feature of breast cancer showed association with the Emax. Breast cancers with higher Emax at shear-wave elastography had poor prognostic features. To some extend, the value of Emax can reflect the biological characteristics of breast cancer, SWE may provide a certain imaging basis for the prognosis of breast cancer and the choice of treatment plan.

Objective To evaluate the value of MicroPure imaging system of ultrasonography in display of breast calcification,and explore its clinical application value.Methods One hundred and forty-one patients underwent by mammography,conventional ultrasound imaging and MicroPure imaging,the mammography results were reference standard,observing the display rate of MicroPure imaging for calcification,and the value of MicroPure imaging in the display of calcification with different shapes on mammography.As the pathologic diagnosis was golden standard,the accuracy of diagnosis of conventional ultrasound,conventional ultrasound combined with MicroPure imaging,conventional ultrasound combined with mammography were evaluated with receiver operating characteristic(ROC) curve.Results The display case of calcifications within breast masses:the specificity of MieroPure imaging was lower than conventional ultrasound(88.14％ vs 100％,P ＜ 0.05),the sensitivity and accuracy were higher than conventional ultrasound(100％ vs 62.79％,93.14％ vs 84.31 ％,P ＜0.05).The display case of microcalcifications within normal breast tissue:the specificity and accuracy of MicroPure software system were lower than conventional ultrasound(9.26％ vs 100％,43.87％ vs 65.96％,P ＜0.05),the sensitivity was higher than conventional ultrasound (100％ vs 20.00％,P ＜0.05).The punctate calcification,amorphous calcification,rough uneven,pleomorphic calcifications on mammography were all hyperechoic spots on MicroPure software system.The area under the ROC curve of the conventional ultrasound,conventional ultrasound combined with MicroPure imaging,conventional ultrasound combined with MicroPure imaging was 0.881,0.858,0.932,respectively.Conclusions MicroPure imaging has some clinical value on microcalcifications within breast masses,which can improve its sensitivity and accuracy.The specificity of MicroPure imaging diagnostic ultrasound imaging of microcalcifications were lower,and it can not distinguish between different shapes of calcifications.Compared with conventional ultrasound,MicroPure imaging system can not improve the accuracy of diagnosis of benign and malignant breast lesions.