Natural Herbal Compounds Exerting an Antidepressant Effect through Hypothalamic-Pituitary-Adrenal Axis Regulation.

Depression is a common mental illness that damages the life and health of patients and causes economic burden, and HPA (hypothalamic-pituitary-adrenal) axis dysfunction is considered to be one of the important factors leading to depression. In this case, it is essential to explore possible treatment methods by using natural compounds with HPA axis regulating and antidepressant effects. However, no one has reviewed it so far. Therefore, the purpose of this review is to systematically sort out the related natural products that play an antidepressant role by regulating the function of the HPA axis. Natural products are divided into flavonoids, polyphenols, terpenoids, saponins, polysaccharides and so on according to their chemical structures, which play a variety of biological activities such as regulating the HPA axis, anti-inflammation and neuroprotection. These effects may provide a useful reference for the potential treatment of depression so as to develop new antidepressants.

Integration of metabolomics and transcriptomics reveals the regulation mechanism of the phenylpropanoid biosynthesis pathway in insect resistance traits in Solanum habrochaites.

Solanum habrochaites (SH), a wild species closely related to 'Ailsa Craig' (AC), is an important germplasm resource for modern tomato breeding. Trichomes, developed from epidermal cells, have a role in defense against insect attack, and their secretions are of non-negligible value. Here, we found that the glandular heads of type VI trichomes were clearly distinguishable between AC and SH under cryo-scanning electron microscopy, the difference indicating that SH could secrete more anti-insect metabolites than AC. Pest preference experiments showed that aphids and mites preferred to feed near AC compared with SH. Integration analysis of transcriptomics and metabolomics data revealed that the phenylpropanoid biosynthesis pathway was an important secondary metabolic pathway in plants, and SH secreted larger amounts of phenylpropanoids and flavonoids than AC by upregulating the expression of relevant genes in this pathway, and this may contribute to the greater resistance of SH to phytophagous insects. Notably, virus-induced silencing of Sl4CLL6 not only decreased the expression of genes downstream of the phenylpropanoid biosynthesis pathway (SlHCT, SlCAD, and SlCHI), but also reduced resistance to mites in tomato. These findings provided new genetic resources for the synthesis of phenylpropanoid compounds and anti-insect breeding in S. habrochaites and a new theoretical basis for the improvement of important traits in cultivated tomato.

A thermal deformation optimization method for cryogenically cooled silicon crystal monochromators under high heat load.

A method to optimize the thermal deformation of an indirectly cryo-cooled silicon crystal monochromator exposed to intense X-rays at a low-emittance diffraction-limited synchrotron radiation source is presented. The thermal-induced slope error of the monochromator crystal has been studied as a function of heat transfer efficiency, crystal temperature distribution and beam footprint size. A partial cooling method is proposed, which flattens the crystal surface profile within the beam footprint by modifying the cooling contact area to optimize the crystal peak temperature. The optimal temperature varies with different photon energies, which is investigated, and a proper cooling strategy is obtained to fulfil the thermal distortion requirements over the entire photon energy range. At an absorbed power up to 300 W with a maximum power density of 44.8 W mm-2 normal incidence beam from an in-vacuum undulator, the crystal thermal distortion does not exceed 0.3 µrad at 8.33 keV. This method will provide references for the monochromator design on diffraction-limited synchrotron radiation or free-electron laser light sources.

20(S)-Ginsenoside Rg2 amino acid derivatives for anti hemorrhagic shock: Synthesis, characterization and evaluation.

The purpose of this study is to screen a novel Rg2 derivative for anti hemorrhagic shock. Eight Rg2 amino acid ester derivatives were designed and synthesized, and their effects on hypoxia and shock were studied. Among them, the derivative 1 (D1) exhibited excellent anti hypoxia by promoting survival rate of H9c2 cells damaged by hypoxia. D1 improved physiological indicators of the rats in hemorrhagic shock, such as blood pressure, heart rate, lactate, acid-base balance, and alleviate oxidative stress and inflammatory damage. Its latent mechanisms were explored by a method of plasma metabolomics based on UPLC-QTOF-MS. As a result, a total of 16 biomarkers were identified involving 6 metabolic pathways. The results of this study contained that the derivative 1 could be considered as potent drug candidates for anti shock and deserved further research and development.

Association of life's essential 8 and risk of fragility fractures: A large cohort study.

Cardiovascular health (CVH) score is not only associated with cardiovascular diseases, but also some disorders in other systems. This study aims to investigate the association between CVH score and the risk of fragility fractures. The analysis enrolled 89,464 participants at baseline in Kailuan study initiated in 2006-2007. All participants were then followed up every 2 years and the incidence of fragility fractures was recorded annually. A total CVH score was classified as low (0-49 points), moderate (50-79 points), and ideal (80-100 points). The primary outcome was incident fragility fractures before December 31, 2021. Kaplan-Meier was used to estimate cumulative incidence. Multivariable adjusted Cox proportional hazards regression models and time-dependent Cox hazards regression models were used to estimate fragility fracture hazard ratios (aHR) and 95% confidence intervals (95% CI). After 13.98 ± 2.84 years of follow-up, a total of 1534 cases of fragility fractures were identified, with an incidence density of 1.23 per 1000 person-years. Compared with the low CVH group, the risk of fragility fractures was significantly lower in moderate (aHR = 0.78, 95% CI: 0.66-0.92) and ideal CVH groups (aHR = 0.65, 95% CI: 0.51-0.83), particularly in the age <60 group (aHR = 0.72, 95% CI: 0.59-0.88; aHR= 0.55, 95% CI: 0.41-0.73, respectively). Time-dependent Cox hazards regression models, sensitivity analysis, and death competition model confirmed the reliability of these findings. The ideal CVH score is associated with a decreased risk of fragility fractures. With the increase of CVH score, the risk of fragility fracture decreases.

Developing a Predictive Model for Minimal or Mild Endometriosis as a Clinical Screening Tool in Infertile Women: Uterosacral Tenderness as a Key Predictor.

STUDY OBJECTIVE: To develop a noninvasive predictive model based on patients with infertility for identifying minimal or mild endometriosis. DESIGN: A retrospective cohort study. SETTING: This study was conducted at a tertiary referral center. PATIENTS: A total of consecutive 1365 patients with infertility who underwent laparoscopy between January 2013 and August 2020 were divided into a training set (n = 910) for developing the predictive model and a validation set (n = 455) to confirm the model's prediction efficiency. The patients were randomly assigned in a 2:1 ratio. INTERVENTIONS: Sensitivities, specificities, area under the curve, the Hosmer-Lemeshow goodness of fit test, Net Reclassification Improvement index, and Integrated Discrimination Improvement index were evaluated in the training set to select the optimum model. In the validation set, the model's discriminations, calibrations, and clinical use were tested for validation. MEASUREMENTS AND MAIN RESULTS: In the training set, there were 587 patients with minimal or mild endometriosis and 323 patients without endometriosis. The combination of clinical parameters in the model was evaluated for both statistical and clinical significance. The best-performing model ultimately included body mass index, dysmenorrhea, dyspareunia, uterosacral tenderness, and serum cancer antigen 125 (CA-125). The nomogram based on this model demonstrated sensitivities of 87.7% and 93.3%, specificities of 68.6% and 66.4%, and area under the curve of 0.84 (95% confidence interval 0.81-0.87) and 0.85 (95% confidence interval 0.80-0.89) for the training and validation sets, respectively. Calibration curves and decision curve analyses also indicated that the model had good calibration and clinical value. Uterosacral tenderness emerged as the most valuable predictor. CONCLUSION: This study successfully developed a predictive model with high accuracy in identifying infertile women with minimal or mild endometriosis based on clinical characteristics, signs, and cost-effective blood tests. This model would assist clinicians in screening infertile women for minimal or mild endometriosis, thereby facilitating early diagnosis and treatment.

MDSCs-derived GPR84 induces CD8+ T-cell senescence via p53 activation to suppress the antitumor response.

BACKGROUNDS: G-protein-coupled receptor 84 (GPR84) marks a subset of myeloid-derived suppressor cells (MDSCs) with stronger immunosuppression in the tumor microenvironment. Yet, how GPR84 endowed the stronger inhibition of MDSCs to CD8+ T cells function is not well established. In this study, we aimed to identify the underlying mechanism behind the immunosuppression of CD8+ T cells by GPR84+ MDSCs. METHODS: The role and underlying mechanism that MDSCs or exosomes (Exo) regulates the function of CD8+ T cells were investigated using immunofluorescence, fluorescence activating cell sorter (FACS), quantitative real-time PCR, western blot, ELISA, Confocal, RNA-sequencing (RNA-seq), etc. In vivo efficacy and mechanistic studies were conducted with wild type, GPR84 and p53 knockout C57/BL6 mice. RESULTS: Here, we showed that the transfer of GPR84 from MDSCs to CD8+ T cells via the Exo attenuated the antitumor response. This inhibitory effect was also observed in GPR84-overexpressed CD8+ T cells, whereas depleting GPR84 elevated CD8+ T cells proliferation and function in vitro and in vivo. RNA-seq analysis of CD8+ T cells demonstrated the activation of the p53 signaling pathway in CD8+ T cells treated with GPR84+ MDSCs culture medium. While knockout p53 did not induce senescence in CD8+ T cells treated with GPR84+ MDSCs. The per cent of GPR84+ CD8+ T cells work as a negative indicator for patients' prognosis and response to chemotherapy. CONCLUSIONS: These data demonstrated that the transfer of GPR84 from MDSCs to CD8+ T cells induces T-cell senescence via the p53 signaling pathway, which could explain the strong immunosuppression of GPR84 endowed to MDSCs.

Effect of skinfold thickness on arm venous access port in cancer patients.

BACKGROUND: Currently, skinfold thickness in studies on arm venous access ports and the effect of venous access port application are unknown. MATERIALS AND METHODS: A total of 256 cancer patients who underwent primary venous access port placement in the Fourth Hospital of Hebei Medical University from September 2022 to March 2023 were selected as the study subjects. Two hundred fifty-six patients were divided into normal skinfold thickness group and high skinfold thickness group according to skinfold thickness. The success rate of primary catheterization of arm venous port catheterization, catheterization operation time, catheterization length and incidence rate of adverse reactions were compared. RESULTS: There was no significant difference in the basic data between the two groups. There was no significant difference in the success rate of primary catheterization between the two groups (p > 0.05), the catheterization operation time in the normal skinfold thickness group was significantly lower than that in the high skinfold thickness group (p < 0.05), the total length of the implanted catheter in the normal skinfold thickness group was significantly lower than that in the high skinfold thickness group (p < 0.05), and the incidence of adverse reactions in the normal skinfold thickness group was significantly lower than that in the high skinfold thickness group (p < 0.05). CONCLUSION: In cancer patients, skinfold thickness can significantly affect the application effect of arm venous port, and normal skinfold thickness for arm venous port has shorter operation time, total length of implanted catheter and lower incidence of adverse reactions.

Putrescine alleviates the oxidative damage of cumulus-oocyte complex via improving fatty acid oxidation.

BACKGROUND: Fatty acid oxidation of cumulus-oocyte complex (COC) provides sufficient energy for oocyte maturation. But, the relationship between fatty acid oxidation and oxidative stress in aging follicles, as well as the effect of putrescine, is still unclear. METHODS: The porcine COCs were randomly divided into four groups and cultured in in vitro maturation (IVM) medium with or without 1 mmol/L putrescine, with 50 µmol/L hydrogen peroxide (H2O2) or with 50 µmol/L H2O2 plus 1 mmol/L putrescine. Oocyte maturation was assessed by the first polar body extrusion. The expressions of genes involved in fatty acid oxidation were detected, and the mitochondrial function was analyzed by themembrane potential. RESULTS: The maturation rate of oocyte was significantly lower in the H2O2 group when compared with the control group (P＜0.001), and putrescine significantly increased this rate in the H2O2 plus putrescine group when compared with the H2O2 group (P＜0.001). The expressions of LKB1, STRAD, ACC2, AMPKα1 and AMPKα2 mRNAs in cumulus cells (CCs) were significantly downregulated by H2O2 treatment, and partly rescued by putrescine addition (P＜0.05-0.001). However, the changes of LKB1, STRAD, ACC2, AMPKα1 and AMPKα2 mRNAs in oocytes were inapparent. The mitochondrial membrane potential of CCs in the H2O2 group was significantly lower than that in the control group, while putrescine addition significantly increased the mitochondrial membrane potential (P＜0.001). CONCLUSION: The decrease of oocyte maturation due to oxidative stress is related with the decreased fatty acid oxidation, and putrescine may alleviate the COCs damage via improving fatty acid oxidation.

The association between blood heavy metals level and sex hormones among postmenopausal women in the US.

Introduction: Environmental pollutants could be implicated in female endocrine setting Q6 beyond traditional factors. Until now, few study has focused on the association of environmental exposure to heavy metals with sex hormones in postmenopausal women. This study intended to investigate whether serum levels of heavy metals(i.e., Cd, Pb, Hg, Mn, Se) would influence sex hormones in postmenopausal women. Methods and results: A cross-sectional study was performed on 614 nationally representative participants from 2013-2016 National Health and Nutrition Examination Survey (NHANES) in the US. Multivariate linear regression models and restricted cubic spline plots revealed cadmium(Cd) had linear positive association with TT(ß=3.25, 95%CI= 1.12, 5.38), bioavailable TT(ß=1.78, 95%CI=0.36,3.21) and TT/E2(ß=0.76, 95%CI=0.28,1.24), which was more apparent in natural menopausal and obese women. Lead(Pb) had linear positive association with SHBG(ß=12.84, 95%CI= 6.77,18.91), which was apparent in nearly all subgroups except in normal BMI group, and TT/E2 (ß=0.69, 95%CI 0.134,1.25), which was apparent in natural menopausal and normal BMI women. Manganese(Mn) had non-linear association with SHBG, which was more apparent in natural menopausal and obese women, and TT/E2, which was more apparent in natural menopausal and normal BMI women. Selenium(Se) had U shaped non-linear association with TT, which was more apparent in hysterectomy, overweight and obese women, and SHBG, which was apparent in nearly all subgroups except in normal BMI group. Conclusion: In summary, this cross-sectional study indicates a possible role that various degree of environmental exposure to heavy metals plays in the disruption of sex Q5 hormone levels in postmenopausal women. Further experiments are needed to elucidate the underlying mechanisms.

Human amnion-derived mesenchymal stem cells improve subclinical hypothyroidism by immunocompetence mediating apoptosis inhibition on thyroid cells in aged mice.

Subclinical hypothyroidism (SCH) affects 10% of the global population, which is most prevalent in women and the elderly. However, it remains debatable whether the elderly with subclinical hypothyroidism needs thyroxine supplement. Human amnion-derived mesenchymal stem cells (hAMSCs) could play important roles in autoimmune diseases, suggesting that hAMSC be a candidate to regulate the thyroid function of female age-related subclinical hypothyroidism. Herein, we established the model of SCH in the aged female mice. This study was designed to investigate whether human amnion-derived mesenchymal stem cells (hAMSC) could effect on immune regulation, apoptosis inhibition of thyroid cells, thyroid function, blood lipid levels, and heart function. In addition, qualified hAMSCs were intravenously injected into aged female SCH mice via the tail vein on day 0 and day 10. The levels of thyroid hormone and blood lipids as well as cardiac function, serum immunological indexes, and apoptosis of thyroid cells were then analyzed on day 5, 10, 15, and 20; meanwhile, the quantity of Th1, Th2, Th17, and Treg immune cells in peripheral blood was evaluated before and on day 20 post-injection. Our study demonstrated that after hAMSC transplantation, the thyroid functions, blood lipid levels, and heart function indexes of age-related SCH (AR-SCH) mice were significantly improved. Consistent with this, Th1 and Treg cells increased significantly, while Th2 and Th17 cells decreased in peripheral blood. Apoptosis was also suppressed in the thyroid cells. In summary, hAMSC delivery can potentially be a safe and effective therapy for treating SCH in the elderly, improving related complications by immunomodulatory and apoptosis inhibition.

Deacetylation of FOXP1 by HDAC7 potentiates self-renewal of mesenchymal stem cells.

BACKGROUND: Mesenchymal stem cells (MSCs) are widely used in a variety of tissue regeneration and clinical trials due to their multiple differentiation potency. However, it remains challenging to maintain their replicative capability during in vitro passaging while preventing their premature cellular senescence. Forkhead Box P1 (FOXP1), a FOX family transcription factor, has been revealed to regulate MSC cell fate commitment and self-renewal capacity in our previous study. METHODS: Mass spectra analysis was performed to identify acetylation sites in FOXP1 protein. Single and double knockout mice of FOXP1 and HDAC7 were generated and analyzed with bone marrow MSCs properties. Gene engineering in human embryonic stem cell (hESC)-derived MSCs was obtained to evaluate the impact of FOXP1 key modification on MSC self-renewal potency. RESULTS: FOXP1 is deacetylated and potentiated by histone deacetylase 7 (HDAC7) in MSCs. FOXP1 and HDAC7 cooperatively sustain bone marrow MSC self-renewal potency while attenuating their cellular senescence. A mutation within human FOXP1 at acetylation site (T176G) homologous to murine FOXP1 T172G profoundly augmented MSC expansion capacity during early passages. CONCLUSION: These findings reveal a heretofore unanticipated mechanism by which deacetylation of FOXP1 potentiates self-renewal of MSC and protects them from cellular senescence. Acetylation of FOXP1 residue T172 as a critical modification underlying MSC proliferative capacity. We suggest that in vivo gene editing of FOXP1 may provide a novel avenue for manipulating MSC capability during large-scale expansion in clinical trials.

Boosting Creativity through Users' Avatars and Contexts in Virtual Environments-A Systematic Review of Recent Research.

As an artificial space extended from the physical environment, the virtual environment (VE) provides more possibilities for humans to work and be entertained with less physical restrictions. Benefiting from anonymity, one of the important features of VEs, users are able to receive visual stimuli that might differ from the physical environment through digital representations presented in VEs. Avatars and contextual cues in VEs can be considered as digital representations of users and contexts. In this article, we analyzed 21 articles that examined the creativity-boosting effects of different digital user and contextual representations. We summarized the main effects induced by these two digital representations, notably the effect induced by the self-similar avatar, Proteus effect, avatar with Social Identity Cues, priming effect induced by contextual representation, and embodied metaphorical effect. In addition, we examined the influence of immersion on creativity by comparing non-immersive and immersive VEs (i.e., desktop VE and headset VE, respectively). Last, we discussed the roles of embodiment and presence in the creativity in VEs, which were overlooked in the past research.

Evaluation of immobilized microspheres of Clonostachys rosea on Botrytis cinerea and tomato seedlings.

Tomato (Solanum lycopersicum L.) is a popular vegetable crop which is widely cultivated around the world. However, the production of tomatoes is threatened by several phytopathogenic agents, including gray mold (Botrytis cinerea Pers.). Biological control using fungal agents such as Clonostachys rosea plays a pivotal role in managing gray mold. However, these biological agents can negatively be influenced by environmental factors. However, immobilization is a promising approach to tackle this issue. In this research, we used a nontoxic chemical material, sodium alginate as a carrier to immobilize C. rosea. For this, sodium alginate microspheres were prepared using sodium alginate prior to embedding C. rosea. The results showed that C. rosea was successfully embedded in sodium alginate microspheres, and immobilization enhanced the stability of the fungi. The embedded C. rosea was able to suppress the growth of gray mold efficiently. In addition, the activity of stress related enzymes, peroxidase superoxidase dismutase and polyphenol oxidation was promoted in tomatoes treated with the embedded C. rosea. By measuring photosynthetic efficiency, it was noted that the embedded C. rosea has positive impacts on tomato plants. Taken together, these results indicate that immobilization of C. rosea improved its stability without detrimentally affecting its efficiency on gray mold suppression and tomato growth. The results of this research can be used as a basis for research and development of new immobilized biocontrol agents.

Fully printed and self-compensated bioresorbable electrochemical devices based on galvanic coupling for continuous glucose monitoring.

Real-time glucose monitoring conventionally involves non-bioresorbable semi-implantable glucose sensors, causing infection and pain during removal. Despite bioresorbable electronics serves as excellent alternatives, the bioresorbable sensor dissolves in aqueous environments with interferential biomolecules. Here, the theories to achieve stable electrode potential and accurate electrochemical detection using bioresorbable materials have been proposed, resulting in a fully printed bioresorbable electrochemical device. The adverse effect caused by material degradation has been overcome by a molybdenum-tungsten reference electrode that offers stable potential through galvanic-coupling and self-compensation modules. In vitro and in vivo glucose monitoring has been conducted for 7 and 5 days, respectively, followed by full degradation within 2 months. The device offers a glucose detection range of 0 to 25 millimolars and a sensitivity of 0.2458 microamperes per millimolar with anti-interference capability and biocompatibility, indicating the possibility of mass manufacturing high-performance bioresorbable electrochemical devices using printing and low-temperature water-sintering techniques. The mechanisms may be implemented developing more comprehensive bioresorbable sensors for chronic diseases.

Therapeutic effects and mechanism of human amnion-derived mesenchymal stem cells on hypercoagulability in a uremic calciphylaxis patient.

Calciphylaxis is a rare cutaneous vascular disease that manifests with intolerable pains, non-healing skin wounds, histologically characterized by calcification, fibrointimal hyperplasia, and microvessel thrombosis. Currently, there are no standardized guidelines for this disease. Recent studies have recognized a high prevalence of thrombophilias and hypercoagulable conditions in calciphylaxis patients. Here, we report a case of uremic calciphylaxis patient whom was refractory to conventional treatments and then received a salvage strategy with intravenous and local hAMSC application. In order to investigate the therapeutic mechanism of hAMSCs from the novel perspective of hypercoagulability, coagulation-related indicators, wound status, quality of life and skin biopsy were followed up. Polymerase chain reaction (PCR) was performed to determine the distribution of hAMSCs in multiple tissues including lung, kidney and muscle after infusion of hAMSCs for 24 h, 1 week and 1 month in mice aiming to investigate whether hAMSCs retain locally active roles after intravenous administration. Improvement of hypercoagulable condition involving correction of platelet, D-dimer and plasminogen levels, skin regeneration and pain alleviation were revealed after hAMSC administration over one-year period. Skin biopsy pathology suggested regenerative tissues after 1 month hAMSC application and full epidermal regeneration after 20 months hAMSC treatment. PCR analysis indicated that hAMSCs were homing in lung, kidney and muscle tissues of mice even until tail vein injection of hAMSCs for 1 month. We propose that hypercoagulability is a promising therapeutic target of calciphylaxis patients, which can be effectively improved by hAMSC treatment.

Alpha-enolase 1 knockdown facilitates the proliferation and invasion of villous trophoblasts by upregulating COX-2.

BACKGROUND: Enolase 1 (ENO1) is a metabolic enzyme which participates in pyruvate synthesis and ATP production in cells. Previously, differential expression of ENO1 was discovered in villous tissues between recurrent miscarriage and induced abortion. This study was designed to explore whether ENO1 influences the proliferation and invasion of villous trophoblasts and the related molecular mechanisms. METHODS: First, ENO1 expression in placental villus tissues collected from recurrent miscarriage (RM) patients and women for induced abortion as well as in trophoblast-derived cell lines was detected by RT-qPCR and western blotting. ENO1 localization and expression in villus tissues were further confirmed through immunohistochemistry staining. Then, the effects of ENO1 downregulation on trophoblast Bewo cell proliferation, migration, invasion, and epithelial-mesenchymal transition (EMT) process were evaluated by CCK-8 assay, transwell assay, and western blotting. As for the regulatory mechanism of ENO1, the expression of COX-2, c-Myc and cyclin D1 in Bewo cells after ENO1 knockdown was finally evaluated by RT-qPCR and western blotting. RESULTS: ENO1 was mainly localized in the cytoplasm, with very small amounts in the nucleus of trophoblast cells. ENO1 expression in the villi tissues of RM patients was significantly increased, when compared with the villous tissues of healthy controls. Furthermore, Bewo cells, a trophoblast cell line with relatively higher expression of ENO1, was used to downregulate the ENO1 expression by ENO1-siRNA transfection. ENO1 knockdown significantly facilitated Bewo cell growth, EMT process, migration, and invasion. ENO1 silencing markedly elevated COX-2, c-Myc, and cyclin D1 expression. CONCLUSION: ENO1 may participate in the development of RM via suppressing the growth and invasion of villous trophoblasts via reducing the expression of COX-2, c-Myc, and cyclin D1.

Development and evaluation of a droplet digital PCR assay to detect Brucella in human whole blood.

BACKGROUND: With the development of domestic animal husbandry, the spread of brucellosis has accelerated, and the scope of the epidemic has expanded. The timely and accurate diagnosis of human brucellosis continues to challenge clinicians in endemic areas. Droplet digital PCR (ddPCR) technology can quickly and accurately determine DNA load in samples, providing laboratory evidence for diagnosis, prognosis and management of brucellosis patients. In this study, a ddPCR method was established to accurately quantify Brucella DNA load in whole blood samples, and its diagnostic, prognostic, and therapeutic value for human brucellosis was evaluated. METHODS: Annealing temperature, primers, and probe targeting the Brucella bcsp31 gene were optimised, and the sensitivity, specificity and repeatability of the ddPCR assay were assessed using 94 whole blood samples from 61 confirmed and 33 suspected cases. Results were compared with those of quantitative PCR (qPCR). Nine follow-up brucellosis patients were also analysed by the two methods after 2 and 6 months of treatment. RESULTS: Optimal primer and probe concentrations were 800 nmol/L and 400 nmol/L, respectively, and the optimal annealing temperature was 55.3 °C. The ddPCR results showed that the limit of detection was 1.87 copies per reaction, with high repeatability. The positive rates for ddPCR and qPCR were 88.5% and 75.4% among 61 serum agglutination test (SAT) positive patients. In addition, 57.6% (19/33) of suspected sero-negative samples were positive by ddPCR, but only 36.3% (12/33) were positive by qPCR. Analysis of nine post-therapy follow-up brucellosis patients revealed that the Brucella DNA load in the whole blood samples decreased after 2 and 6 months of treatment, and was slightly increased following relapse and continuous exposure. CONCLUSION: The ddPCR assay showed good accuracy for whole blood samples, and could be a potential diagnostic and prognostic tool for detecting Brucella.

Efficacy of dehydroepiandrosterone priming in women with poor ovarian response undergoing IVF/ICSI: a meta-analysis.

Background: Dehydroepiandrosterone (DHEA) may improve the outcomes of patients with poor ovarian response (POR) or diminished ovarian reserve (DOR) undergoing IVF/ICSI. However, the evidence remains inconsistent. This study aimed to investigate the efficacy of DHEA supplementation in patients with POR/DOR undergoing IVF/ICSI. Methods: PubMed, Web of Science, Cochrane Library, China National Knowledge Infrastructure (CNKI) were searched up to October 2022. Results: A total of 32 studies were retrieved, including 14 RCTs, 11 self-controlled studies and 7 case-controlled studies. In the subgroup analysis of only RCTs, DHEA treatment significantly increased the number of antral follicle count (AFC) (weighted mean difference : WMD 1.18, 95% confidence interval(CI): 0.17 to 2.19, P=0.022), while reduced the level of bFSH (WMD -1.99, 95% CI: -2.52 to -1.46, P<0.001), the need of gonadotropin (Gn) doses (WMD -382.29, 95% CI: -644.82 to -119.76, P=0.004), the days of stimulation (WMD -0.90, 95% CI: -1.34 to -0.47, P <0.001) and miscarriage rate (relative risk : RR 0.46, 95% CI: 0.29 to 0.73, P=0.001). The higher clinical pregnancy and live birth rates were found in the analysis of non-RCTs. However, there were no significant differences in the number of retrieved oocytes, the number of transferred embryos, and the clinical pregnancy and live birth rates in the subgroup analysis of only RCTs. Moreover, meta-regression analyses showed that women with lower basal FSH had more increase in serum FSH levels (b=-0.94, 95% CI: -1.62 to -0.25, P=0.014), and women with higher baseline AMH levels had more increase in serum AMH levels (b=-0.60, 95% CI: -1.15 to -0.06, P=0.035) after DHEA supplementation. In addition, the number of retrieved oocytes was higher in the studies on relatively younger women (b=-0.21, 95% CI: -0.39 to -0.03, P=0.023) and small sample sizes (b=-0.003, 95% CI: -0.006 to -0.0003, P=0.032). Conclusions: DHEA treatment didn't significantly improve the live birth rate of women with DOR or POR undergoing IVF/ICSI in the subgroup analysis of only RCTs. The higher clinical pregnancy and live birth rates in those non-RCTs should be interpreted with caution because of potential bias. Further studies using more explicit criteria to subjects are needed. Systematic review registration: https://www.crd.york.ac.uk/prospero/, identifier CRD 42022384393.