Wnt3a-induced LRP6 phosphorylation enhances osteoblast differentiation to alleviate osteoporosis through activation of mTORC1/ß-catenin signaling.

OBJECTIVE: Osteoporosis (OP) is a common cause of morbidity and mortality in older individuals. The importance of Wnt3a in osteogenic activity and bone tissue homeostasis is well known. Here, we explored the possible molecular mechanism by which Wnt3a mediates the LRP6/mTORC1/ß-catenin axis to regulate osteoblast differentiation in OP. METHODS: OP-related key genes were identified through a bioinformatics analysis. A ROS17/2.8 cell differentiation system for rat osteogenic progenitors and a rat model of senile OP were constructed for in vitro and in vivo mechanism verification. RESULTS: Bioinformatics analysis revealed that LRP6 was poorly expressed in OP and may play a key role in the occurrence of OP by affecting osteoblast differentiation. LRP6 knockdown inhibited osteoblast differentiation in an in vitro model. In addition, Wnt3a promoted osteoblast differentiation by inducing LRP6 phosphorylation. Moreover, LRP6 promoted mTORC1 expression, which indirectly promoted ß-catenin expression, thus promoting osteoblast differentiation. Finally, an in vivo assay revealed that LRP6 inhibition improved OP. CONCLUSION: Our study provides evidence that Wnt3a induces phosphorylation of LRP6 to activate the mTORC1/ß-catenin axis, thus promoting osteoblast differentiation and ultimately improving OP in aged rats.

Determination of 3-chloro-1,2-propanediol in biological samples by quick-easy-cheap-effective-rugged-and-safe extraction coupled with gas chromatography-mass spectrometry and its biodistribution in rats.

3-Chloro-1,2-propanediol is a common food contaminant, but reports on its determination in biological tissues are lacking. In the present study, a method was developed to detect 3-chloro-1,2-propanediol contents in rat tissues by quick-easy-cheap-effective-rugged-and-safe extraction and gas chromatography-mass spectrometry analysis. Biological samples were extracted with ethyl acetate and purified with adsorbents. The optimized adsorbent for each sample was selected from 4-5 combinations of N-propylethylenediamine, octadecylsilane, graphitized carbon black, strong anion exchange, and florisil. Extracted 3-chloro-1,2-propanediol was derivatized with heptafluorobutyric anhydride and subjected to gas chromatography-mass spectrometry. This method had good linearity (correlation coefficients >0.99) in the range of 2-2000 ng/g for blood, kidney, liver, testis, and brain samples. The limits of detection were under 0.8 ng/g; the limits of quantification were 2 ng/g; the recovery rates were 85%-102%; and the matrix effects were 1.98%-7.67%. This method also had good precision. The dynamic changes in 3-chloro-1,2-propanediol in rats gavaged with 20 mg/kg b.w. for 24 h were detected using this method. The 3-chloro-1,2-propanediol content in each tissue sharply increased to a peak, rapidly decreased within 2 h, and stabilized at 12 h. 3-Chloro-1,2-propanediol persisted in the kidney, testis, and liver 24 h after gavage.

Ferulic Acid Induces Autophagy and Apoptosis in Colon Cancer CT26 Cells via the MAPK Pathway.

Ferulic acid (FA) is a bioactive compound found in traditional Chinese herbal medicine; for example, it is present in Xinjiang Ferula, but also in strong-flavor Chinese baijiu. FA has been shown to play a crucial role in treating oxidative stress, skin whitening, and eye diseases. In this study, the potential role of FA as a means of inducing apoptosis and inhibiting colon cancer induced by the transplantation of CT26 cells was investigated. The results show that FA adjuvant treatment caused an upregulation in the expression of genes related to autophagy while simultaneously suppressing the expression of inflammatory response elements and improving the bodyweight, glutamic pyruvic transaminase (ALT), and glutamic oxaloacetic transaminase (AST) in vivo. Furthermore, FA inhibited the proliferation of CT26 cells and induced apoptosis, specifically by activating the phosphorylation of ERK and JNK to enhance the essential proteins BCL-2 and BAX in the apoptosis pathway. These results suggest that FA could be a promising auxiliary therapeutic agent for the treatment of colon cancer. Further research is needed to better understand the mechanisms underlying the beneficial effects of FA and its synergistic effects with other compounds.

Survival of Lactobacillus casei strain Shirota in the intestines of healthy Chinese adults.

Lactobacillus casei strain Shirota (LcS) is a widely used probiotic strain with health benefits. In this study, the survival of LcS in the intestines of healthy Chinese adults was assessed and the effects of LcS on stool consistency, stool SCFAs and intestinal microbiota evaluated. Subjects consumed 100 mL per day of a probiotic beverage containing 1.0 × 10(8) CFU/mL of LcS for 14 days. LcS were enumerated using a culture method and the colony identity confirmed by ELISA. Fourteen days after ingestion, the amount of LcS recovered from fecal samples was between 6.86 ± 0.80 and 7.17 ± 0.57 Log10 CFU/g of feces (mean ± SD). The intestinal microbiotas were analyzed by denaturing gradient gel electrophoresis. Principal component analysis showed that consuming LcS significantly changed fecal microbiota profiles. According to redundancy analysis, the amounts of 25 bacterial strains were significantly correlated with LcS intake (P < 0.05), 11 of them positively and fourteen negatively. Concentrations of acetic acid and propionic acid in feces were significantly lower during the ingestion period than during the baseline period (P < 0.05). These results confirm that LcS can survive passage through the gastrointestinal tract of Chinese people; however, they were found to have little ability to persist once their consumption had ceased. Furthermore, consumption of probiotic beverages containing LcS can modulate the composition of the intestinal microbiota on a long-term basis, resulting in decreased concentrations of SCFAs in the gut.

Bisphenol A Exposure Interferes with Reproductive Hormones and Decreases Sperm Counts: A Systematic Review and Meta-Analysis of Epidemiological Studies.

Bisphenol A (BPA), an acknowledged endocrine disrupter, is easily exposed to humans via food packaging and container. However, a consensus has not been reached on the extent to which BPA exposure affects the reproductive system. We therefore conducted this systematic review and meta-analysis to elucidate the relationship between BPA exposure and male reproduction-related indicators. Up to October 2023, a comprehensive search was carried out in the PubMed, Embase, Cochrane and Web of Science, and 18 studies were ultimately included. ß coefficients from multivariate linear regression analyses were pooled using a random effects model. The results showed that the urinary BPA concentration was negatively correlated with the sperm concentration (ß coefficient = -0.03; 95% CI: -0.06 to -0.01; I2 = 0.0%, p = 0.003) and total sperm count (ß coefficient = -0.05; 95% CI: -0.08 to -0.02; I2 = 0.0%, p < 0.001). In addition, BPA concentrations were associated with increased sex hormone-binding globulin (SHBG) levels, increased estradiol (E2) levels, and reduced biologically active androgen levels. However, the relationship between an increased risk of below-reference sperm quality and BPA exposure was not robust. This systematic review revealed that BPA exposure disrupts reproductive hormones, reduces sperm counts and may ultimately adversely affect male reproduction.

Lactobacillus paracaseiN1115 attenuates obesity in high-fat diet-induced obese mice.

Disruption of the microbial structure of intestinal bacteria due to a high-fat diet (HFD) is closely associated with metabolic disorders, such as obesity and type 2 diabetes. Probiotics are known to modulate the gut microbiota; therefore, we demonstrated the capability of Lactobacillus paracasei N1115 (LC-N1115) to attenuate obesity. Four-week-old male C57BL/6J mice were fed a HFD for 12 weeks to induce obesity and were then randomized to supplemented placebo or LC-N1115 treatment group for another 12 weeks. LC-N1115 treatment reduced weight gain and liver fat accumulation as well as triglyceride, total cholesterol, and low-density lipoprotein cholesterol levels. The administration of LC-N1115 suppressed the expression of fatty acid synthase, interleukin-1 ß, and toll-like receptor 4. Notably, the operational taxonomic units that negatively and positively correlated with the obesity phenotypes were enriched and reduced, respectively, in the LC-N1115 treatment group. These results indicate that LC-N1115 attenuates obesity by modulating the gut microbiota and the expression of lipid synthesis and proinflammatory cytokine genes.

Lactobacillus paracasei L9 ameliorated obesity-associated metabolic parameters and relevant gut microbiota in mice fed a high-fat diet.

The purpose of the present study was to determine whether Lactobacillus paracasei L9 (L9) supplementation prevents diet-induced obesity in C57BL/6J mice. Four-week-old mice were fed a high-fat diet (HFD) for 12 weeks and then supplemented with or without L9 for another 12 weeks. Weight gain, white adipose tissue weight, plasma lipid levels of total cholesterol, triglyceride, and low-density lipoprotein-cholesterol were significantly increased in the HFD group compared with those in the control group and were decreased by L9 treatment. The fat deposits in the liver and epididymal adipose tissue were increased in the HFD group compared with the normal chow diet group and decreased by L9 treatment. Reverse transcriptase-polymerase chain reaction analyses revealed that L9 suppressed pro-inflammatory cytokine and lipid synthesis-related genes in epididymal adipose tissue. This study used Illumina Miseq sequencing to explore alterations of the gut microbiome. L9 ameliorated HFD-induced structural dysbiosis and gut bacteria that were positively related with obesity phenotypes were obviously decreased. Altogether, the findings indicate that administration of L9 ameliorates HFD-induced hyperlipidemia and lipid accumulation in liver and inflammation associated with intestinal dysbiosis in obese mice. These findings suggest that L9 supplementation may provide a natural alternative to attenuate obesity.

3-Monochloropropane-1,2-diol causes spermatogenesis failure in male rats via Sertoli cell dysfunction but not testosterone reduction.

3-Monochloropane-1,2-diol (3-MCPD), a common food contaminant, has been confirmed to impair male fertility, but the mechanism has not been fully clarified. This study systematically explored the spermatogenesis impairment induced by 3-MCPD in vivo and in vitro with a focus on Sertoli cells (SCs) and spermatogonial stem cells (SSCs). After adult male Sprague-Dawley rats were administered 36 and 72 mg/kg b.w./day 3-MCPD daily for 4 weeks, the total sperm concentration dramatically decreased by 28.9 % and 57.7 %, respectively, and obvious testicular seminiferous tubule atrophy was observed. 3-MPCD exposure decreased serum testosterone levels but not intratesticular testosterone levels and upregulated the expression of steroidogenesis enzymes in both rat testes and primary Leydig cells. 3-MCPD did not reduce the number and self-renewal marker PLZF+ of SSCs; however, it downregulated the key meiotic genes Stra8 and Rec8 in the rat testis but not in primary germ cells. Although SC counts were not affected, 3-MCPD downregulated androgen receptor (AR) in rat testes and primary SCs. In addition, 3-MCPD downregulated p-CREB (transcription factor of AR), paracrine meiosis regulators Nrg1 and Nrg3 and retinoic acid synthetase Aldh1a1 in primary SCs. In summary, 3-MCPD caused impairment of spermatogenesis by inhibiting secretion of meiosis regulators and disturbing testosterone signalling in SCs.

Effects of oligosaccharides on the markers of glycemic control: a systematic review and meta-analysis of randomized controlled trials.

Objective: To investigate the effect of oligosaccharides on the markers of glycemic control, including fasting blood glucose (FBG), fasting blood insulin (FBI), glycated hemoglobin (HbA1c), homeostasis model assessment of insulin resistance (HOMA-IR), and quantitative insulin sensitivity index (QUICKI). Methods: PubMed, Embase, and the Cochrane Library databases were systematically searched to find randomized controlled trials (RCTs) on the effect of oligosaccharide intervention on FBG, FBI, HbA1c, HOMA-IR, and QUICKI up to 7 June 2021. Data were pooled using weighted mean difference (WMD) and 95% confidence intervals (95% CI), with a p-value ≤0.05 indicating statistical significance. Risk of bias was assessed with the Cochrane tool and the quality of the literature with the new Jadad scale. Results: A total of 46 randomized controlled trials were included. Oligosaccharides significantly reduced FBG (WMD: -0.295 mmol L-1; 95% CI: -0.396 to -0.193; p < 0.001; I2 = 90.9%; 46 trials; 2412 participants), FBI (WMD: -0.559 pmol L-1; 95% CI: -0.939 to -0.178; p < 0.01; I2 = 99.1%; 29 trials; 1462 participants), HbA1c (WMD: -0.365; 95% CI: -0.725 to -0.005; p < 0.05; I2 = 86.6%; 11 trials; 661 participants), and HOMA-IR (WMD: -0.793; 95% CI: -1.106 to -0.480; p < 0.001; I2 = 96.1%; 24 trials; 1382 participants). Oligosaccharides were more beneficial for the participants with obesity or diabetes than for healthy participants. Multiple interventions per day consolidated the effectiveness of oligosaccharides. Regardless of the processing manner (starch-modified or naturally extracted) of the oligosaccharides, their intervention was overall beneficial for the patients with diabetes. Conclusions: This study is by far the most extensive systematic review to evaluate the role of oligosaccharides on the markers of glycemic control. Oligosaccharide interventions can exert beneficial effects on FBG, FBI, HbA1c, and HOMA-IR.

The effects of biogenic amines in Chinese Huangjiu on the behavior of mice and hangover headache-related indices.

Huangjiu (Chinese rice wine) is a popular and traditional alcoholic beverage in China; however, the consumption of Huangjiu readily results in hangover symptoms. The aim of this study was to identify the main components associated with behavioral inhibition, headache, and the relevant mechanisms by using a mice hangover model. The results of an open-field experiment revealed that the key biogenic amine associated with mice behavior was histamine, which inhibited the behavior activity of mice in a dose-dependent manner. Moreover, histamine treatment decreased the levels of serotonin (5-HT) and 5-hydroxyindole acetic acid. In addition, the levels of dopamine and nitric oxide, which are associated with migraine, increased in the brain tissue of mice. In addition, the expression of receptor genes of 5-HT, including Htr1a, Htr1f, and Htr2c, is essential in regulating various behaviors and mental activities. In conclusion, the present study demonstrated that histamine is a key component in Huangjiu, and it is related to hangover symptoms by affecting the level of 5-HT and its receptors.

Effects of Fermented Milk Containing Lacticaseibacillus paracasei Strain Shirota on Constipation in Patients with Depression: A Randomized, Double-Blind, Placebo-Controlled Trial.

Probiotics have been shown to benefit patients with constipation and depression, but whether they specifically alleviate constipation in patients with depression remains unclear. The aim of this study was to investigate the effect of Lacticaseibacillus paracasei strain Shirota (LcS), formerly Lactobacillus casei strain Shirota, on constipation in patients with depression with specific etiology and gut microbiota and on depressive regimens. Eighty-two patients with constipation were recruited. The subjects consumed 100 mL of a LcS beverage (108 CFU/mL) or placebo every day for 9 weeks. After ingesting beverages for this period, we observed no significant differences in the total patient constipation-symptom (PAC-SYM) scores in the LcS group when compared with the placebo group. However, symptoms/scores in item 7 (rectal tearing or bleeding after a bowel movement) and items 8-12 (stool symptom subscale) were more alleviated in the LcS group than in the placebo group. The Beck Depression Index (BDI) and Hamilton Depression Rating Scale (HAMD) scores were all significantly decreased, and the degree of depression was significantly improved in both the placebo and LcS groups (p < 0.05), but there was no significant difference between the groups. The LcS intervention increased the beneficial Adlercreutzia, Megasphaera and Veillonella levels and decreased the bacterial levels related to mental illness, such as Rikenellaceae_RC9_gut_group, Sutterella and Oscillibacter. Additionally, the interleukin (IL)-1ß, IL-6, and tumor necrosis factor-α (TNF-α) levels were significantly decreased in both the placebo and LcS groups (p < 0.05). In particular, the IL-6 levels were significantly lower in the LcS group than the placebo group after the ingestion period (p < 0.05). In conclusion, the daily consumption of LcS for 9 weeks appeared to relieve constipation and improve the potentially depressive symptoms in patients with depression and significantly decrease the IL-6 levels. In addition, the LcS supplementation also appeared to regulate the intestinal microbiota related to mental illness.

Differential Effects of Lactobacillus casei Strain Shirota on Patients With Constipation Regarding Stool Consistency in China.

BACKGROUND/AIMS: Probiotics are expected to confer benefits on patients with constipation, but how probiotics act on constipated patients with variable stool consistencies remains unclear. We investigated the effect of Lactobacillus casei strain Shirota (LcS) on constipation-related symptoms, especially stool consistency, of constipated patients. METHODS: Constipated patients meeting the Rome III criteria were divided into 3 groups according to the Bristol Stool Form Scale (BSFS): hard (hard stool [HS], BSFS ＜ 3), normal (normal stool [NS], ≤ 3 BSFS ≤ 4), and soft (soft stool [SS], 4 ＜ BSFS ≤ 5) stools. Subjects in each group consumed a probiotic beverage containing 10¹° colony-forming units of LcS daily for 28 days. RESULTS: LcS intervention significantly alleviated constipation-related symptoms and increased defecation frequency in all subjects. Four weeks of LcS supplementation softened the hard stools in HS, hardened the soft stools in SS, and did not alter the ideal stool consistency in NS. The short-chain fatty acid (SCFA) concentrations were highest in SS, followed by NS and HS. LcS intervention increased the stool SCFA levels in HS but reduced or did not alter the levels in NS and SS. LcS intervention increased the Pseudobutyrivibrio and Roseburia abundances in HS and decreased the Pseudobutyrivibrio abundance in SS. CONCLUSIONS: LcS supplementation improved the constipation-related symptoms in constipated subjects. Differences in baseline stool consistency could result in different anti-constipation effects of LcS intervention. LcS balanced the stool consistency-softened the HS and hardened the SS. These effects could be associated with modulation of the gut microbiota and SCFA production.

Lactobacillus casei Strain Shirota Alleviates Constipation in Adults by Increasing the Pipecolinic Acid Level in the Gut.

The benefits of probiotics for constipation are widely accepted, but the mechanisms involving gut metabolites are unclear. In this study, we investigated the effects of Lactobacillus casei strain Shirota (LcS) on constipated patients and revealed that a metabolite mediator is involved in the LcS-induced constipation alleviation. Sixteen constipated patients and 22 non-constipated participants were recruited. The subjects consumed 100 mL of an LcS beverage (108 CFU/mL) per day for 28 days. The fecal non-volatile metabolites were determined by GC/MS, and the targeted metabolites were further verified in a constipated mouse model. In constipated patients, LcS intervention significantly improved defecation frequency (from 4.81 to 7.81 times per week, p < 0.05), stool consistency (from 2.52 to 3.68, p < 0.05) and constipation-related symptoms. A total of 14 non-volatile fecal metabolites were obtained as potential constipation-related metabolites that were regulated by LcS. Among these metabolites, pipecolinic acid (PIPA) had a significant positive correlation with defecation frequency in constipated patients. PIPA significantly promoted the small intestinal propulsive rate (from 25.45 to 39.68%) and increased the number of fecal pellets (from 30.38 to 57.38 pellets) in constipated mice (p < 0.05). The 5-hydroxytryptamine (5-HT) and acetylcholine (ACh) in colonic tissue may be partly involved in PIPA-mediated constipation alleviation. In conclusion, PIPA was a metabolic mediator in the gut that participated in LcS-induced constipation alleviation.

[Comparison of clinical outcome between cemented and biological femoral prosthesis for the treatment of senile femoral neck fracture].

OBJECTIVE: To study and compare the early fixed effect and mortality for cemented and biological femoral prosthesis in treatment of senile osteoporotic femoral neck fracture, and investigate the prosthesis choice in hip arthroplasty. METHODS: From January 2012 to December 2014, 130 patients(130 hips) with osteoporotic femoral neck fracture were treated by hemiarthroplasty. According to the operation method the patients was divided into two groups: 72 cases in cemented group included 26 males and 46 females with an average age of (82.0±6.5) years old, 32 cases were Garden III and 40 cases were Garden IV, the average time from injury to operating was(5.5±3.3) days; 58 cases in biological group included 19 males and 39 females with an average age of(80.1±6.7) years old, 21 cases were Garden III and 37 cases were Garden IV, and the average time from injury to operating was(5.4±2.1) days. Every patients were suffering from hip swelling, tenderness, axial taps lower limb pain and joint activities obstacles. ASA grade evaluation, heart function assessment, surgical treatment and postoperative follow-up effectively performed after admission. The operation duration, intraoperative blood loss, hospital stay, rate of complications, mortality, residual pain (VAS score) and hip function (Harris score) of two groups were observed and compared. RESULTS: Biological group received shorter operation time and less intraoperative blood loss than cemented group(P<0.05). Perioperative mortality was 2.7%(2 patients) in cemented group, and no one death in biological group. Finally, 128 patients were follow-up for an average of 22 months. Complications and VAS score of cemented group was lower than that of biological group during follow-up(P<0.05), but Harris score was higher (P<0.05). There was no statistical difference in mortality between two groups during follow-up period(P>0.05). CONCLUSIONS: The early fixed effect for the cemented prosthesis used in treatment of senile osteoporotic femoral neck fracture is better than biological stem. Some patients who with higher ASA grade or poor cardiopulmonary function can choose biological prosthesis in order to reduce perioperative mortality.

Changes in the microbiota of lamb packaged in a vacuum and in modified atmospheres during chilled storage analysed by high-throughput sequencing.

Changes in the microbiota of lamb were investigated under vacuum packaging (VP) and under 20% CO2/80% N2 (LC), 60% CO2/40% N2 (MC), and 100% CO2 (HC) modified atmosphere packaging (MAP) during chilled storage. Viable counts were monitored, and the total microbial communities were assessed by high-throughput sequencing. The starting community had the highest microbial diversity, after which Lactococcus and Carnobacterium spp. outcompeted during the 28-day storage. The relative abundances of Brochothrix spp. in the LC atmosphere were much higher than those of the other groups on days 7 and 28. The bacterial inhibiting effect of the MAP environments on microbial growth was positively correlated with the CO2 concentration. The HC atmosphere inhibited microbial growth and delayed changes in the microbial community composition, extending the lamb's shelf life by approximately 7days compared with the VP atmosphere. Lamb packaged in the VP atmosphere had a more desirable colour but a higher weight loss than lamb packaged in the MAP atmospheres.