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: He-Wei Granule (HWKL) is a modern product derived from the modified formulation of traditional Chinese medicine Banxia Xiexin Decoction (BXD), which remarkedly enhanced the anti-proliferation activity of cyclophosphamide (CTX) on HepG2 and SGC-7901 cell lines in vitro in our previous research. The aim of the study was to investigate the synergistic effects of HWKL and CTX using a transplanted H22 hepatocellular carcinoma mouse model. Methods: The CTX-toxic-reducing efficacy of HWKL was evaluated by hematology indexes, organ indexes and marrow DNA detection. To investigate the underlying mechanisms, histopathology test, immunohistochemistry test and TUNEL staining were conducted. The efficacy of HWKL on the micro-vessel density (MVD) in tumor tissue was also evaluated by measuring CD34 level. Results: High dose HWKL (6.75 g/kg) markedly attenuated CTX-induced hepatotoxicity and myelosuppression while significantly enhanced CTX anticancer efficacy in vivo. Further mechanism investigation suggested that high dose HWKL significantly increased cleaved Caspase 3 level and promoted apoptosis in tumor tissue by up-regulating Bax expression and down-regulating Bcl-2 and FasL expressions. Compared with CTX alone group, the decrease in LC-3B and Beclin 1 levels suggested that the autophagy in H22 carcinoma was significantly inhibited with addition of high dose HWKL. ELISA assay results indicated that the autophagy inhibition was achieved by decreasing p53 expression, blocking PI3K/AKT/mTOR pathway and recovering Th1/Th2 cytokine balance. In addition, CD34 and EGFR immunohistochemistry assay suggest that high dose HWKL could significantly decrease micro-vessel density (MVD) and inhibit angiogenesis in H22 carcinoma. Conclusion: It can be concluded that high-dose HWKL enhanced CTX efficacy by promoting apoptosis, inhibiting autophagy and angiogenesis in tumor tissue while significantly alleviated CTX-induced toxicity, and could be applied along with CTX in clinical treatment as a supplement agent.

Objective:To investigate the effect of gene polymorphism of β1 adrenergic receptor (β1-AR) G1165C and A145G locus on myocardial hypertrophy and the efficacy in patients with hypertension.Methods:Two hundred and twenty-seven cases of patients with hypertension admitted to Binhai County People′s Hospital from January to December 2019 were enrolled. Among them, there were 113 cases of hypertension with myocardial hypertrophy and 114 cases of hypertension without myocardial hypertrophy. In addition, 115 patients with normal physical examination during the same period were selected as the control group. DNA in the peripheral blood leukocytes was extracted, polymerase chain reaction-restriction fragment length polymorphism method was used to detect β1-AR G1165C and A145G locus gene polymorphism, and the differences in the efficacy of β blockers in hypertensive patients with different genotypes were compared.Results:There was no statistically significant differences in the distribution of β1-AR A145G genotypes among the three groups ( P>0.05). Compared with the healthy control group, the frequency of Gly/Gly genotype carrying β1-AR G1165C locus was higher in hypertension with myocardial hypertrophy group, and the frequency of Gly/Arg and Arg/Arg gene were lower; compared with hypertension without myocardial hypertrophy group, the frequency of Gly/Arg+Gly/Gly gene in hypertension with myocardial hypertrophy group was higher; taking Arg/Arg genotype as the control group, carrying Gly/Gly genotype could increase the risk of cardiac hypertrophy in hypertensive patients by 3.159 times ( OR = 3.159, 95% CI 1.240 - 7.412, P<0.05).The frequency of G1165C allele Arg in the hypertension with myocardial hypertrophy group was significantly lower than that in the control group and the hypertension without myocardial hypertrophy group ( P<0.05); the frequency of G1165C allele Gly was significantly higher than that in the control group and the hypertension without myocardial hypertrophy group ( P<0.05); taking Arg/Arg genotype as the control, carrying Gly/Gly genotype could increase the risk of cardiac hypertrophy in hypertensive ( OR = 3.417, 95% CI 1.357 - 7.965, P<0.05). The left ventricular mass index of Gly/Gly genotype patients was (120.38 ± 28.41) g/m 2, which was significantly higher than (99.76 ± 25.16) g/m 2 and (90.30 ± 19.54) g/m 2 of Gly/Arg and Arg/Arg, with statistically significant differences ( F = 10.89, P<0.01). After the treatment, the resting heart rate, systolic blood pressure, diastolic blood pressure and mean arterial blood pressure of patients with G1165C allele Arg hypertension with myocardial hypertrophy were lower than those with G1165C allele Gly, with statistically significant differences ( P<0.05). Conclusions:β1-AR G1165C gene polymorphism is related to the risk of myocardial hypertrophy in hypertensive patients. Carrying the G1165C allele Gly may increase the risk of susceptibility to cardiac hypertrophy, and β-blockers are more effective in hypertensive patients with myocardial hypertrophy who carry the G1165C allele Arg.

OBJECTIVE：To investigate the risk factors for the recurrence of ischemic stroke after secondary prevention ，and to observe the effect of glutathione on 4-HNE. METHODS ：Totally 97 patients with ischemic stroke relapse within one year were treated from Oct. 2017 to Oct. 2019 in 3 hospitals as the Second Affiliated Hospital of Shandong First Medical University due to cerebral thrombosis or cerebral embolism as observation group ，and 97 non-recurrence patients in the same period were paired as control group. The patients in the observation group were randomly divided into conventional treatment group （49 cases）and drug intervention group （48 cases）. The patients in conventional treatment group received routine treatment such as cerebral blood flow recanalization， improving circulation ， controlling blood pressure ， maintaining blood glucose ， treating hyperlipidemia and arrhythmia during hospitalization. Drug intervention group was additionally given Glutathione for injection 1.8 g intragastrically ， once a day ，on the basis of conventional treatment group. 4-HNE concentrations in plasma were determined at admission and 14 days after treatment ，the genetic type of ALDH2 and type of TAST were determined at admission. Multiple liner regression was used to analyze the factors associated with 4-HNE increasing ； conditional Logistic analysis was used to identify independent risk factors resulting to ischem ic stroke recurrence after secondary prevention. RESULTS ：The plasma concentration of 4-HNE at admission and the percentage of arte ry atherosclerosis patients in observation group were significantly higher than control group（P＜0.05）. The distribution of each ALDH2 genotype in 2 groups complied with Hardy-Weinberg genetic equilibrium （P＞ 0.05）. The proportion of patients carrying ALDH2*2 allele in observation group （50.50％）was significantly higher than control group（36.08％）（P＜0.05）. ALDH2*2 allele [ B＝2.33，95％CI（1.35，5.50），P＝0.03] and artery atherosclerosis [ B＝1.90，95％CI （1.29，3.74），P＝0.04] were significantly associated with the elevation of plasma concentration of 4-HNE；artery atherosclerosis [OR＝ 2.93，95％CI（1.84，4.67），P＜0.01]，stroke family history [OR ＝1.50，95％CI（1.18，1.90），P＝0.04]，elevated plasma concentration of 4-HNE [OR ＝1.34，95％CI（1.11，1.62），P＝0.04] were regarded as independent risk factors associating with ischemic stroke recurrence after secondary prevention. After intervention ，plasma concentration of 4-HNE in drug intervention group and conventional treatment group was significantly lower than before intervention （P＜0.05）；there was no statistical significance between 2 groups（P＞0.05）. CONCLUSIONS ：Stroke family history ，artery atherosclerosis and the elevation plasma concentration of 4-HNE are independent risk factors associating with ischemic stroke recurrence after secondary prevention. Although drug intervention can reduce the elevated plasma concentration of 4-HNE，the effect of additional use of glutathione is not more significant than that of conventional treatment.

Objective: To perform lymphocyte micronucleus analysis on radiation workers with long-term exposure to low doses ionizing radiation, Evaluate the health condition of radiation workers, and provide the evidence for strengthening surveillance of radiation workers. Methods: From January 1, 2013 to December 21, 2016, a statistical analysis and evaluation was conducted of the peripheral lymphocytes micronucleus rate in 5 901 radiation workers who had undergone medical examinations of employees at Chinese Academy of Medical Sciences Institute of Radiation Medicine. Results: The micronucleus rates in radiation workers of the on-job group were higher than the pre-job group (P<0.01) . Significant difference was found among the different sex (t=5.97) , different types (χ²=378.69) , different levels of work units (χ²=115.48) . Significant difference was found among the micronucleus rates of 672 radiation workers of the on-job group from 2013 to 2016 (χ²=92.57, P<0.01) . Conclusion The peripheral lymphocytes micronucleus rate of radiation workers were significantly higher than non-contact workers. Significant increasing trend of micronucleus rates was noted among the radiation worker with increasing exposure time. The peripheral lymphocytes micronucleus rates of interventional therapy workers were highest. The peripheral lymphocytes micronucleus rates of Private hospitals workers were highest. This phenomenon deserves attention. Protection needs to be strengthened to ensure the health of radiation workers.

Objective:To investigate the prevalence,and the factors that influence depressive symptoms among chronic respiratory disease patients in tertiary hospital.Methods:A total of 1713 outpatients and inpatients with chronic respiratory disease were selected from 8 tertiary hospitals in Jiangsu Province from July to September,2014 and screened according to the Hospital Anxiety Depression Scale-D (HADS-D).A questionnaire developed by this research group,was used to collect demographic and clinical information.Logistic regression was used to identify factors that were associated with depressive symptoms.Results:The overall rate of depressive symptoms was 46.0％.Multiple logistic analysis showed that spinsterhood (OR ＝ 0.45),higher education level (middle school /high school/technical school OR ＝0.65;college degree or aboveOR ＝0.28),BMI ≥24 (OR ＝0.71) were associated with decreased risk of depressive symptoms (P ＜ 0.05).B MI ＜ 18.5 (OR ＝ 1.52,),average income of family ≥10000 RMB (OR ＝ 1.37-1.96),limited daily activities (OR ＝ 1.72),poorer sleep quality (OR ＝ 1.45),and negative life events (OR ＝ 1.62) were associated with increased risk of depressive symptoms (P ＜ 0.05).Conclusion:The prevalence of depressive symptoms among chronic respiratory disease patients in tertiary hospitals in Jiangsu Province was higher.Marital status,education level,income,BMI,limited daily activities,subjective sleep quality,negative life events may be the related factors of depressive symptoms of chronic respiratory diseases patients.

BACKGROUND:In view of the unavoidable problems of autogenous and al ogenous bone grafts, it is an urgent need to develop biodegradable bone substitute materials, among which is calcium phosphate material that has become a hot spot in the domestic and foreign research. OBJECTIVE:To develop a biodegradable calcium phosphate material for bone repair based on tetracalcium phosphate (TTCP). METHODS:The biodegradable calcium phosphate cement made from TTCP, dicalcium phosphate anhydrous and different constituents of curing liquids was prepared under room temperature (about 25 ℃). The effects of solid components, liquid components as well as calcinations and drying temperature on the physical and biological performances were detected through X-ray diffraction test, hardness test, decay in a simulated body fluid, hemolysis and cytotoxicity tests, respectively, to select the bone repair material with excel ent performances. RESULTS AND CONCLUSION:When the calcination temperature was lower than 1300 ℃, TTCP was rarely available;only close to 1400 ℃, the relatively pure TTCP was gained. A large number of pure TTCP were gained by rapid cooling because of avoidance of the moisture impact, but slow cooling made the main products to be hydroxyapatite, suggesting that rapid cooling is essential to obtain pure TTCP. With the increase of the proportion of citric acid solution in the liquid phase, the pH values and the hemolysis rate in the bone cement soak solution were increased gradually, illustrating that citric acid solution is easy to induce hemolysis. In vitro cel experiments showed that the hemolysis rate of bone cement with a solution of 2:1 NH4/Na ratio was the lowest, and the biocompatibility was the best, which was the most favorable to cel growth. Cements was made of calcined powders sieved at 1400 ℃ and showed the shortest initial setting time, least effect on pH values, lowest disintegration rate and hemolysis rate, and slightest inhibition effect on the cel proliferation, indicating that the bone cements made of sieved powder after 1400 ℃ calcination is more suitable for clinical application.

Objective To evaluate the short term clinical efficacy of recombinant human B‐type natriuretic peptide(rhBNP) in the treatment of dilated cardiomyopathy complicating heart failure .Methods 121 patients with dilated cardiomyopathy complicating heart failure were selected ,the cardiac function grade Ⅲ - Ⅳ ,and randomly divided into the conventional treatment group(control group ,n= 61) and the rhBNP treatment group(rhBNP group ,n = 60) .The disease history was recorded and clinical symptoms , heart color echocardiography ,cardiac function ,renal function and plasma NT‐proBNP levels were observed before and after treat‐ment .Results The NT‐proBNP level after 72 h treament in the rhBNP group was significantly lower than that in the control group (P< 0 .01) ;LVEDd after 1 week treatment in the rhBNP group was significantly lower than that in the control group ( P =0 .033) ;LVEF was increased in the both groups ,but the increase in the rhBNP group was more significant compared with the con‐trol group (P< 0 .01) .The total effective rate was 91 .6% in the rhBNP group and 72 .1% in the control group with statistical dif‐fernece between the two groups(P= 0 .005) ;the average hospital stay time in the rhBNP group was significantly shorter than that in the control group(P= 0 .041) .The proportion of the major adverse cardiovascular events(MACE) occurrence had no statistical difference between the two groups(P= 0 .492) .Conclusion rhBNP is safe and effective in treating the acute decompensation of di‐lated cardiomyopathy .

Graphene/Room temperature ionic liquids ( GN/IL ) nanocomposite was prepared by grinding graphene and ionic liquid 1-butyl-3-methylimidazolium bromide ( BMIMPF6 ) which mixed together in appropriate proportion. Atomic force microscopy ( AFM ) was utilized to characterize the formation of the GN-ILs. Due to the synergistic effect between ionic liquids and graphene, the nanocomposite exhibited excellent performance toward H2 O2 reduction. A novel uric acid ( UA) electrochemical sensor was fabricated based on uricase-GN/IL modified glassy carbon electrode. The experimental results showed that the response displayed a good linear response toward UA in the concentration range from 0. 002-4. 5 mmol/L. The corre1ation coefficient was 0. 995 and the detection limit was 0. 85 μmol/L. The easily prepared electrochemical sensor had favorable stability and selectivity and could be applied to the quick determination of UA in human serum, thus providing a new UA detection method for clinical trial.