Role of miR-300-3p in Leydig cell function and differentiation: A therapeutic target for obesity-related testosterone deficiency.

MicroRNAs (miRNAs) regulate various cellular functions, but their specific roles in the regulation of Leydig cells (LCs) have yet to be fully understood. Here, we found that the expression of miR-300-3p varied significantly during the differentiation from progenitor LCs (PLCs) to adult LCs (ALCs). High expression of miR-300-3p in PLCs inhibited testosterone production and promoted PLC proliferation by targeting the steroidogenic factor-1 (Sf-1) and transcription factor forkhead box O1 (FoxO1) genes, respectively. As PLCs differentiated into ALCs, the miR-300-3p expression level significantly decreased, which promoted testosterone biosynthesis and suppressed proliferation of ALCs by upregulating SF-1 and FoxO1 expression. The LH/METTL3/SMURF2/SMAD2 cascade pathway controlled miR-300-3p expression, in which luteinizing hormone (LH) upregulated SMAD-specific E3 ubiquitin protein ligase 2 (SMURF2) expression through methyltransferase like 3 (METTL3)-mediated Smurf2 N6-methyladenosine modification. The Smurf2 then suppressed miR-300 transcription by inhibiting SMAD family member 2 (SMAD2) binding to the promoter of miR-300. Notably, miR-300-3p was associated with an obesity-related testosterone deficiency in men and the inhibition of miR-300-3p effectively rescued testosterone deficiency in obese mice. These findings suggested that miR-300-3p plays a pivotal role in LC differentiation and function, and could be a promising diagnostic or therapeutic target for obesity-related testosterone deficiency.

Danggui-Shaoyao-San Promotes Amyloid-ß Clearance through Regulating Microglia Polarization via Trem2 in BV2 Cells.

BACKGROUND: Alzheimer's disease (AD) is a chronic neurodegenerative brain disorder currently without satisfactory therapeutic treatments. Triggering receptors expressed on a myeloid cells-2 (Trem2) gene mutation has been reported as a powerful AD risk factor that induces Trem2 gene deletion aggravated microglia disfunction and Amyloid-ß (Aß) aggregation in the brain. The traditional Chinese medicine (TCM) formula Danggui-Shaoyao-San (DSS) has shown therapeutic effect on alleviating the symptoms of AD. However, the neuroprotective effect and underlying mechanism of DSS against AD is still far from fully understood. METHODS: Double-label immunofluorescence and Western blotting were employed to evaluate the different polarization states of mouse BV2 microglial (BV2) cells after lipopolysaccharide (LPS) or interleukin (IL)-4 treatment. Trem2 over-expression lentiviral vector and Trem2 siRNA were used respectively to evaluate the effect of Trem2 on microglia polarization via detecting the proteins expression of iNOS and arginase1 (Arg1) by Western blotting while the Aß-scavenging capacity of BV2 cells was assessed by flow cytometry. Cell counting kit-8 (CCK8) assay was performed to assess the effect of DSS on the viability of BV2 cells. Flow cytometry was used to investigate the effect of DSS on the Aß-scavenging capacity of BV2 cells treated with corresponding concentration of DSS-containing serum. Protein of Trem2 and the gene expression of the M1 or M2 phenotype in BV2 cells treated with DSS after Trem2 over-expression or silence were detected by Western blot and RT-qPCR, respectively. RESULTS: In vitro experiments. DSS exhibited anti-inflammatory and neuroprotective functions. It was found that Trem2 had an effect on inducing a shift of M1 microglia towards the M2 phenotype and enhanced the Aß-scavenging capacity of BV2 cells, further that DSS administration relieved inflammation by engulfing Aß through the activities of Trem2. Importantly, DSS treatment effectively increased the Aß-scavenging capacity of BV2 cells through accelerating the shift of M1 microglia towards an M2 phenotype via increasing Trem2 expression. CONCLUSIONS: Results demonstrated that DSS promoted the clearance of Aß through the regulation of microglia polarization via increased expression of Trem2 in BV2 cells.

Treatment Progress in Diminished Ovarian Reserve: Western and Chinese Medicine.

Diminished ovarian reserve (DOR), generally defined as a decreased number or quality of oocytes, has a significant impact on quality of life and fertility in women. In recent years, the incidence of DOR has been increasing and the ages of patients are younger. The search for an effective DOR treatment has emerged as one of the preeminent research topics in reproductive health. An effective DOR therapy would improve ovarian function, fertility, and quality of life in patients. In this review we evaluated DOR treatment progress both in Western medicine and Chinese medicine, and elucidated the characteristics of each treatment.

Recombinant Photolyase-Thymine Alleviated UVB-Induced Photodamage in Mice by Repairing CPD Photoproducts and Ameliorating Oxidative Stress.

Cyclobutane pyrimidine dimers (CPDs) are the main mutagenic DNA photoproducts caused by ultraviolet B (UVB) radiation and represent the major cause of photoaging and skin carcinogenesis. CPD photolyase can efficiently and rapidly repair CPD products. Therefore, they are candidates for the prevention of photodamage. However, these photolyases are not present in placental mammals. In this study, we produced a recombinant photolyase-thymine (rPHO) from Thermus thermophilus (T. thermophilus). The rPHO displayed CPD photorepair activity. It prevented UVB-induced DNA damage by repairing CPD photoproducts to pyrimidine monomers. Furthermore, it inhibited UVB-induced ROS production, lipid peroxidation, inflammatory responses, and apoptosis. UVB-induced wrinkle formation, epidermal hyperplasia, and collagen degradation in mice skin was significantly inhibited when the photolyase was applied topically to the skin. These results demonstrated that rPHO has promising protective effects against UVB-induced photodamage and may contribute to the development of anti-UVB skin photodamage drugs and cosmetic products.

In vitroreconstitution of the hormone-responsive testicular organoids from murine primary testicular cells.

Increasing rates of male infertility require more experimental models to understand the mechanisms underlying male infertility.In vitroorganoids hold unprecedented promise for this purpose; however, the development of organoids with tissue architecture similar to that of the testisin vivoremains a challenge. Here, we generated testicular organoids derived from testicular cells by combining a hanging drop culture and a rotation culture system. Our results indicated that testicular cells could self-assemble into spheroid organoids with tubule-like structures in hanging drop culture. The organoids can subsequently be cultured and maintained in a rotation culture system. These established organoids have gene expression profiles similar to those of adult testis tissue, produce testosterone with preserved gonadotropin responsiveness, and exhibit sensitivity to reproductive toxicants. More importantly, each testicular organoid can be generated from only 2000 cells, and they maintain their proliferative ability after freezing and thawing. These features make it possible to obtain fresh primary testis cells from testicular biopsies taken from patients or endangered wild species, and to build individual-specific biobanks. These findings will help enable the exploration of self-organization process of testicular cells and provide an experimental model for reproductive biology research, pharmacotoxicology testing, and regenerative medicine.

Small-Molecule-Driven Direct Reprogramming of Fibroblasts into Functional Sertoli-Like Cells as a Model for Male Reproductive Toxicology.

Sertoli cells (SCs) are vital to providing morphological and nutritional support for spermatogenesis. Defects in SCs often lead to infertility. SCs transplantation is a promising potential strategy to compensate for SC dysfunction. However, isolation of SCs from testes is impractical due to obvious and ethical limitations. Here, a molecular cocktail is identified comprising of pan-BET family inhibitor (I-BET151), retinoic acid, and riluzole that enables the efficient conversion of fibroblasts into functional Sertoli-like cells (CiSCs). The gene expression profiles of CiSCs resemble those of mature SCs and exhibit functional properties such as the formation of testicular seminiferous tubules, engulfment of apoptotic sperms, supporting the survival of germ cells, and suppressing proliferation of primary lymphocytes in vitro. Moreover, CiSCs are sensitive to toxic substances, making them an alternative model to study the deleterious effects of toxicants on SCs. The study provides an efficient approach to reprogram fibroblasts into functional SCs by using pure chemical compounds.

Low-Frequency Magnetic Stimulation of Shenmen Acupoint Reduces Blood Oxygen Levels in the Prefrontal Cortex of Healthy Subjects: A Near-Infrared Brain Functional Imaging Study.

OBJECTIVE: To explore the effect of low-frequency magnetic stimulation at Shenmen (HT 7) acupoint on blood oxygen levels in the prefrontal cortex of healthy subjects. METHODS: Functional near-infrared spectroscopy (fNIRS) technology was used to collect real-time data of oxygenated hemoglobin (oxy-Hb) in the prefrontal cortex of 16 healthy subjects at resting state and low-frequency magnetic stimulation of Shenmen. The mean and integral values of blood oxygen concentration were analyzed. RESULTS: Compared with the resting state, the mean and integral values of blood oxygen concentration were decreased during the task period, recovery period, and the whole process in the magnetic stimulation of Shenmen acupoint (P<0.05). In particular, the difference was statistically significant in the recovery period (P<0.01). CONCLUSIONS: The prefrontal cortex was widely activated and produced an immediate effect by reducing the local blood oxygen concentration at low-frequency magnetic stimulation of Shenmen acupoint, which verifies the sedative effect of Shenmen acupoint.

Effect of magnetic stimulation of Shenmen point on cognitive function of chronic insomnia: A randomized controlled clinical trial.

BACKGROUND: Chronic insomnia (CI) can lead to cognitive dysfunction and bring great pain to patients' life. There is no effective intervention for cognitive dysfunction caused by CI. Shenmen (HT7) is the first choice for insomnia treatment. However, the effect and mechanism of this acupoint on cognitive function after insomnia is not clear. Therefore, the purpose of this study is to explore whether magnetic stimulation of HT7 can improve cognitive impairment of CI by regulating prefrontal lobe and its mechanism. METHODS/DESIGN: This is a randomized controlled clinical trial. Seventy-two subjects aged 18 to 65 years old with primary insomnia and more than 3 months were randomly divided into 2 groups according to the ratio of 1:1, and 36 healthy controls were included. The control group was given sleep hygiene and cognitive therapy in behavioral cognitive therapy technology, while the experimental group was given the behavioral cognitive therapy technology intervention and magnetic stimulation of HT7 acupoint for 30 times (2 times / d, 5 times / wk for 20 days), while the healthy control group had no intervention measures. Before treatment and 20 days after treatment, we evaluated the working memory (1-back test), episodic memory (Complex Figure Test), and problem-solving ability (Hanoi tower test) processed by prefrontal lobe to explore the effect of magnetic stimulation on cognitive function of CI and its possible mechanism. At the same time, insomnia severity index was used to evaluate sleep state, Becker depression scale was used to evaluate depression, and Beck anxiety scale was used to evaluate anxiety. Chi-squared test or rank sum test was used to collect the data of patients. If P value is less than or equal to .05, the difference will be considered statistically significant. CONCLUSION: This study explored the effect and mechanism of magnetic stimulation of Shenmen (HT7) on cognitive function of CI, and confirmed that magnetic stimulation of HT7 can be used as an alternative therapy to improve cognitive impairment of CI. TRIAL REGISTRATION NUMBER: ChiCTR2000034280.

How Can Manufacturers Promote Green Innovation in Food Supply Chain? Cost Sharing Strategy for Supplier Motivation.

In the innovation of production activities by green product manufacturing or application, food supply chain cooperation is an important method to optimize the allocation of internal and external innovation resources, strengthen their own core capabilities and achieve sustainable development of enterprises. Whether the traditional revenue sharing or cost sharing strategy is still efficient in the food supply chain cooperation aiming at green innovation attracts a lot of attention. Further research about whether the traditional cooperation contract can effectively motivate suppliers to maximize their innovation efforts is required. In this paper, the green innovation effort level parameters are designed and the constraint factor of the green preference of consumers at the market end is applied to discuss the incentive strategy of cost sharing led by manufacturers. Stackelberg equilibrium structure is utilized in the incentive model in this paper to discuss the existence of the optimal cost sharing ratio, the optimal effort level and the optimal income of green innovation cooperation in the food supply chain. The results show that when the supply is interrupted due to the insufficient stimulation of green consumption at the market demand side, manufacturers need to stimulate their green innovation efforts by sharing the cost of suppliers, and the cost sharing proportion is affected by the marginal profit coefficient of manufacturers and suppliers. When the relationship between the marginal profit of suppliers and the marginal profit of manufacturers reaches a certain threshold, manufacturers use the cost sharing contract, which can effectively stimulate the green innovation efforts of suppliers and optimize the overall income of the food supply chain.

Observation for the effect of rTMS combined with magnetic stimulation at Neiguan (PC6) and Sanyinjiao (SP6) points on limb function after stroke: A study protocol.

BACKGROUND: Stroke is the primary cause of adult disability in China, which causes serious personal, family, and social burden. "Central peripheral central" closed-loop rehabilitation theory is proved to be an effective neural rehabilitation model. Based on this theory, repetitive transcranial magnetic stimulation (rTMS) combined with magnetic stimulation of Neiguan (PC6) and Sanyinjiao (SP6) may be an effective treatment for limb dysfunction after stroke. However, the efficacy and mechanism of repetitive magnetic stimulation of M1 region combined with magnetic stimulation of Neiguan and Sanyinjiao points on limb dysfunction after stroke has not been confirmed. METHODS/DESIGN: This study is a prospective, randomized, controlled, open trial. We randomly divided 42 subjects, aged 35 to 80 years, diagnosed with ischemic stroke within 1 month, into 2 groups with a ratio of 1:1. On the basis of this medical treatment, patients in the experimental group received 1 Hz rTMS in M1 area on the contralateral side, and 3 Hz rTMS treatment at Neiguan point and Sanyinjiao point on the affected side. The control group was treated with acupuncture (body acupuncture). All patients were treated once a day and followed up for 10 days. The National Institute of Health Stroke Scale score, simplified fulg Meyer, modified Barthel index, and cortical excitability were evaluated on the day of enrollment and the 10th day of treatment respectively. The modified Barthe index was followed up on the 30th day of treatment, and the adverse reactions were recorded at any time. The mechanism of rTMS will be revealed by Barthe index before treatment, on the 10th day of treatment and on the 30th day of follow-up. The results were analyzed by spss19.0 software, and the quantitative indexes were analyzed by t test and rank sum test. χ test was used for non-grade counting, and rank sum test was used for grade counting. All statistical tests were performed with bilateral test. If P value is less than or equal to .05, the difference will be considered statistically significant. CONCLUSION: The purpose of this study was to determine the effect of repetitive magnetic stimulation of M1 region combined with magnetic stimulation of Neiguan and Sanyinjiao points on limb function after stroke. Through this study, we expect to explore a new scheme for the treatment of poststroke dyskinesia, and prove that compared with rTMS and acupuncture alone, the closed-loop rehabilitation theory based on "center peripheral center" can be more efficient and safe in the treatment of poststroke limb dysfunction. TRIAL REGISTRATION: The trial was registered in China clinical trial registry (http://www.chictr.org.cn/index.aspx), ID: ChiCTR1900026890 (October 25, 2019).

Effect of low frequency repetitive magnetic stimulation at Shenmen (HT7) on sleep quality in patients with chronic insomnia.

BACKGROUND: Insomnia is a common, recurrent, and tenacious sleep problem, especially the chronic insomnia. Repetitive transcranial magnetic stimulation (rTMS) at right dorsolateral prefrontal cortex (r-DLPFC) is used in chronic insomnia, and repetitive magnetic stimulation (rMS) at Shenmen (HT7) acupoint may be an alternative approach. However, the efficacy and mechanism of rMS at HT7 acupoint for chronic insomnia has not been confirmed. METHODS/DESIGN: This is a 3-armed randomized positive-controlled noninferiority clinical trial. We will allocate 45 subjects aged between 18 and 65 years old, diagnosed with initial chronic insomnia over 3 months to 3 groups randomly in a ratio of 1:1:1. Patients in the experimental group will be treated with rMS at HT7 acupoint while the others in the control group will be treated with rTMS at r-DLPFC or waiting treatment. All will be given rMS at HT7 or rTMS at r-DLPFC or no treatment for 10 days, and then received 20-day follow-up. Patients will be evaluated with the insomnia severity index and Pittsburgh sleep quality index for sleep state, Beck Depression Inventory-2nd edition scores for the depression state, Beck anxiety inventory scores for the anxiety state, and Montreal Cognitive Assessment scores for the cognition state before and the 10th day of treatment, 30th day of follow-up; study on mechanisms of rMS will be revealed through the resting motor threshold diversity of the motor cortex before and the 10th day of treatment, 30th day of follow-up. Baseline characteristics of patients will be summarized by groups and compared with Chi-squared for categorical variables, and analysis of variance or Kruskal-Wallis test for the continuous variables. Primary and secondary outcomes according to the measurement times are applicable to univariate repetitive measurement deviation analysis or analysis of variance, or Kruskal-Wallis test. CONCLUSION: The present study is designed to preliminarily investigate short-term efficacy and mechanism of rMS at HT7 acupoint therapy on chronic insomnia, also to explore the correlation between motor cortex excitability and chronic insomnia. With this research, we are looking forward to find out an appropriate alternative and easy therapy for chronic insomnia individuals compared with rTMS at r-DLPFC. TRIAL REGISTRATION: The trial was registered on Chinese Clinical Trial Registry (http://www.chictr.org.cn/index.aspx) with the ID ChiCTR1900026844 on October 24, 2019.

Association analysis between vitamin D level and depression in women perimenopause: A protocol of systematic review and meta-analysis.

BACKGROUND: In recent decades, many researches manifested that the perimenopause is a window of vulnerability for the development of both depressive symptoms and major depressive episodes. Some scholar thought that those women diagnosed with depression may be particularly sensitive to changes in the hormonal milieu experienced premenstrual, during the postpartum period or during the menopause transition in. Risk factors for depressive symptoms during the perimenopause include prior standardized mean difference (MDD), psychosocial factors, anxiety symptoms, and reproductive-related mood disturbance. However, active vitamin D (VD), exerts protective and regulatory effects on the brain dopamine system and suggests that similar to the antidepressant. Therefore, serum 25(OH)D level may be negatively correlated with the perimenopausal depression. METHODS: The study only selects clinical randomized controlled trials of depression in perimenopausal women. We will search each database from the built-in until October 2020. The English literature mainly searches Cochrane Library, PubMed, EMBASE, and Web of Science. While the Chinese literature comes from CNKI, CBM, VIP, and Wangfang database. Meanwhile, we will retrieve clinical trial registries and grey literature. Two researchers worked independently on literature selection, data extraction, and quality assessment. The dichotomous data is represented by relative risk, and the continuous is expressed by mean difference or standard mean difference, eventually the data is synthesized using a fixed effect model or a random effect model depending on the heterogeneity. The serum vitamin D level, Hamilton Depression Scale, or Beck Depression Inventory or Zung self-rating depression scale or patient health questionnare-9 were evaluated as the main outcomes. While several secondary outcomes were also evaluated in this study. The statistical analysis of this Meta-analysis was conducted by RevMan software version 5.3. RESULTS: This meta-analysis will further determine the association analysis between VD level and depression in women perimenopause. CONCLUSION: This study determines the VD level is related to the occurrence of depression in perimenopausal women.

Do Consumer's Green Preference and the Reference Price Effect Improve Green Innovation? A Theoretical Model Using the Food Supply Chain as a Case.

Today, complex consumer purchase decisions affect company revolution, especially that dealing with environmental (green) innovation. Consumer preferences and pricing could interact in influencing green innovation. This situation is especially vital in industries that are characterized with many safety concerns that urge collaborative development between consumers and companies, such as the food industry. The aim of this paper is to explore the influence of the change in consumer preference characteristics on the green innovation efforts of the food supply chain, introducing the green preference of consumers and the reference price effect. A differential game model of green innovation in the food supply chain is constructed in this paper and solved. It was found that the change in consumer preference characteristics is an important factor to motivate supply chain members to make green innovation efforts. With the enhancement of consumer preference characteristics, the Pareto improvement effect of cost-sharing contracts on the profits of supply chain enterprises is clearer. Further, manufacturers can stimulate suppliers' green innovation efforts through cost-sharing contracts. When the marginal profit ratio of food supply chain members reaches a certain threshold, the incentive effect of this cost-sharing contract is more significant, and it is more likely to realize the optimal profit of the food supply chain.

[Clinical thinking about treating acute ischemic stroke by targeting the neurovascular unit of Chinese medicine].

Neurovascular unit (NVU) concept proposed for the treatment of acute ischemic stroke (AIS) provides a new target, i.e., we should target as an integrity including neurons, glia, and microcirculation, thus supplementing limitations of previous treatment targeting neurons or blood vessels alone. Meanwhile, many clinical trials have failed after NVU protection against AIS drug research has developed at home and abroad. Chinese medicine has multi-component, multi-target, and overall regulation advantages, and is in line with clinical requirement for overall treatment targeting multiple targets of NVU. Currently clinical studies of Chinese medicine treatment of AIS targeting NVU are few. Standardized and systematic clinical efficacy evaluation is lack. Clinical studies for improving AIS-NVU injured blood markers by Chinese medicine are rarer. We hope to pave the way for performing clinical studies on Chinese medicine treatment of AIS targeting NVU.