Mechanistic study of the anti-excitatory amino acid toxicity of Bushen Zhichan decoction for Parkinson's disease based on the transcriptional regulation of EAAT1 by YY1.

ETHNOPHARMACOLOGICAL RELEVANCE: Bushen Zhichan decoction (BSZCF) is derived from Liuwei Dihuang Pill, a famous Chinese herbal formula recorded in the book Key to Therapeutics of Children's Diseases. It has been widely used as a basic prescription for nourishing and tonifying the liver and kidneys to treat Parkinson's disease (PD), but its mechanism remains to be explored. AIM OF THE STUDY: BSZCF, a Chinese herbal formula comprising five herbs: Rehmannia glutinosa (Gaertn.) DC., Dioscorea oppositifolia L., Cornus officinalis Siebold & Zucc., Fallopia multiflora (Thunb.) Haraldson and Cistanche tubulosa (Schenk) Wight, is used clinically to treat PD. In vivo and in vitro experiments were designed to elucidate the mechanism of BSZCF in the protection of dopamine (DA) neurons and the treatment of PD. The toxicity of excitatory amino acids (EAA) may be attenuated by inhibiting the transcription factor Yin Yang 1 (YY1) and up-regulating the expression of excitatory amino acid transporter 1 (EAAT1). MATERIALS AND METHODS: IN VIVO: After 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) was intraperitoneally injected into specific pathogen free (SPF) C57BL/6J mice, model mice were intragastrically given adamantane hydrochloride tablets (AHT) or different doses of BSZCF for 14 days. Both open field and pole-climbing tests were conducted to assess behavioral changes. In vitro: 1-Methyl-4-phe-nylpyridiniumiodide (MPP+)-injured human neuroblastoma cells (SH-SY5Y) were utilized to construct PD cell models. Primary astrocytes were transfected with EAAT1 and YY1 lentiviruses for EAAT1 gene knockout and YY1 gene knockout astrocytes, respectively. The high performance liquid chromatography-mass spectrometry (HPLC-MS) analysis of BSZCF was performed to control the quality of blood drugs. The optimal concentration and time of PD cell models treated by BSZCF were determined by the use of Cell Counting Kit-8 (CCK8). Enzyme-linked immunosorbent assay (ELISA) was used for measuring glutamate (Glu) in the peripheral blood and cells of each group. Western blotting (WB) and real-time quantitative polymerase chain reaction (qPCR) were used to detect tyrosine hydroxylase (TH), dopamine transporters (DAT), EAAT1 and YY1 protein and mRNA. After the blockade of EAAT1, immunofluorescence (IF) assay was used to detect the TH protein in each group. RESULTS: In vivo research showed that BSZCF improved the behavioral symptoms of PD mice, and reduced the death of DA neurons and the level of Glu. The mechanism may be related to the decrease of YY1 expression and the increase of EAAT1 levels. In vitro experiments showed that the anti-excitatory amino acid toxicity of BSZCF was achieved by inhibiting YY1 expression and regulating EAAT1. CONCLUSIONS: By inhibiting YY1 to increase the expression of EAAT1 and attenuating the toxicity of Glu, BSZCF exerts the effect of protecting DA neurons and treating PD-like symptoms in mice.

New Insights into Mechanisms Traditional Chinese Medicine for Allergic Rhinitis by Regulating Inflammatory and Oxidative Stress Pathways.

Allergy rhinitis (AR) is becoming more common and has serious medical and societal consequences. Sneezing, paroxysmal nasal blockage, nasal itching, mucosal edema, coughing, and rhinorrhea are symptoms of this type I allergic immunological illness. Immunoglobulin E-mediated inflammation is the cause of it. Because AR is prone to recurrent attacks, extended medication therapy may impair its effectiveness. In addition to negatively affecting the patients' physical health, this can also negatively impact their mental health. During AR development, there are inflammatory and oxidative stress responses that are linked to problems in a number of signal transduction pathways. By using the terms "allergic rhinitis", "traditional Chinese medicine", "inflammation", and "oxidative stress", we screened for pertinent research published over the previous five years in databases like PubMed. We saw that NF-KB, TLR, IL-33/ST2, PI3K/AKT, MAPK, and Nrf2 are some of the most important inflammatory and oxidative stress pathways in AR. Studies have revealed that antioxidant and anti-inflammatory therapy reduced the risk of AR and was therapeutic; however, the impact of the therapy varies widely. The Chinese medical system places a high value on traditional Chinese medicine (TCM), which has been there for virtually all of China's 5000-year history. By influencing signaling pathways related to inflammation and oxidative stress, Chinese herbal medicine and its constituent compounds have been shown to prevent allergic rhinitis. This review will focus on this evidence and provide references for clinical treatment and scientific research applications.

Metabolomics: A useful tool for ischemic stroke research.

Ischemic stroke (IS) is a multifactorial and heterogeneous disease. Despite years of studies, effective strategies for the diagnosis, management and treatment of stroke are still lacking in clinical practice. Metabolomics is a growing field in systems biology. It is starting to show promise in the identification of biomarkers and in the use of pharmacometabolomics to help patients with certain disorders choose their course of treatment. The development of metabolomics has enabled further and more biological applications. Particularly, metabolomics is increasingly being used to diagnose diseases, discover new drug targets, elucidate mechanisms, and monitor therapeutic outcomes and its potential effect on precision medicine. In this review, we reviewed some recent advances in the study of metabolomics as well as how metabolomics might be used to identify novel biomarkers and understand the mechanisms of IS. Then, the use of metabolomics approaches to investigate the molecular processes and active ingredients of Chinese herbal formulations with anti-IS capabilities is summarized. We finally summarized recent developments in single cell metabolomics for exploring the metabolic profiles of single cells. Although the field is relatively young, the development of single cell metabolomics promises to provide a powerful tool for unraveling the pathogenesis of IS.

FHL3 promotes the formation of fast glycolytic muscle fibers by interacting with YY1 and muscle glycolytic metabolism.

The proportions of the various muscle fiber types are important in the regulation of skeletal muscle metabolism, as well as animal meat production. Four-and-a-half LIM domain protein 3 (FHL3) is highly expressed in fast glycolytic muscle fibers and differentially regulates the expression of myosin heavy chain (MyHC) isoforms at the cellular level. Whether FHL3 regulates the transformation of muscle fiber types in vivo and the regulatory mechanism is unclear. In this study, muscle-specific FHL3 transgenic mice were generated by random integration, and lentivirus-mediated gene knockdown or overexpression in muscles of mice or pigs was conducted. Functional analysis showed that overexpression of FHL3 in muscles significantly increased the proportion of fast-twitch myofibers and muscle mass but decreased muscle succinate dehydrogenase (SDH) activity and whole-body oxygen consumption. Lentivirus-mediated FHL3 knockdown in muscles significantly decreased muscle mass and the proportion of fast-twitch myofibers. Mechanistically, FHL3 directly interacted with the Yin yang 1 (YY1) DNA-binding domain, repressed the binding of YY1 to the fast glycolytic MyHC2b gene regulatory region, and thereby promoted MyHC2b expression. FHL3 also competed with EZH2 to bind the repression domain of YY1 and reduced H3K27me3 enrichment in the MyHC2b regulatory region. Moreover, FHL3 overexpression reduced glucose tolerance by affecting muscle glycolytic metabolism, and its mRNA expression in muscle was positively associated with hemoglobin A1c (HbA1c) in patients with type 2 diabetes. Therefore, FHL3 is a novel potential target gene for the treatment of muscle metabolism-related diseases and improvement of animal meat production.

A clinical nomogram based on absolute count of lymphocyte subsets for predicting overall survival in patients with non-small cell lung cancer.

BACKGROUND: The absolute count of lymphocyte subsets (ACLS) is correlated to the prognosis of multiple malignancies. This study aimed to combine the ACLS with the clinicopathological parameters to develop a nomogram to accurately predict the prognosis of non-small cell lung cancer (NSCLC) patients. METHODS: This retrospective study included a training cohort (n = 1685) and validation cohort (n = 337) with NSCLC patients treated in First Teaching Hospital of Tianjin University of Traditional Chinese Medicine between January 2018 and January 2021. Cox regression were conducted to identify factors associated with overall survival. The nomogram was built based on 10 significant factors, and evaluated by the concordance index (C-index), calibration curve and receiver operating characteristic (ROC) curve. RESULTS: In the training cohort, the multivariate cox proportional hazard regression analysis showed that the independent factors for overall survival (OS) included age, brain metastases, hepatic metastases, respiratory system diseases, clinical stages, surgery, absolute count (AC) of CD3+, CD4+, CD8+, and NK cells, which were all applied in the nomogram. The C-index of the nomogram to predict OS was 0.777 (95% CI, 0.751-0.802) in training cohort and 0.822 (95% CI, 0.798-0.846) in validation cohort. The area under the ROC showed a good discriminative ability in both cohorts. Calibration curves presented an excellent consistence between the nomogram predicted probability and actual observation. CONCLUSIONS: We established a prognostic nomogram to predict OS of the NSCLC patient. This nomogram provided a more quantitative, scientific and objective basis for accurate diagnosis and individual management of NSCLC patients.

Observation of the Curative Effect of Acupuncture for Tonifying Kidney and Removing Blood Stasis Combined with Radiofrequency Surgery in Patients with NSCLC and the Diagnostic Efficacy of Combined Detection of NTx, BGP, and CYFRA21-1 in the Occurrence of Bone Metastases.

The curative effect observation of acupuncture for tonifying kidney and removing blood stasis combined with radiofrequency surgery in patients with non-small-cell lung carcinoma (NSCLC) and the diagnostic efficacy of combined detection of NTx, BGP, and CYFRA21-1 for bone metastases are investigated. 122 NSCLC patients admitted to our hospital from January 2019 to December 2021 are selected for the examination, and the two sets of patients are randomly divided into the study set and the control set using the random number table method, with 61 cases in each set. Patients in the control set are given CT-guided percutaneous radiofrequency ablation therapy, and patients in the study set are given a combination of acupuncture therapy for tonifying the kidney and removing blood stasis on the basis of the therapy of the control set. The experimental results show that for NSCLC patients, the application of kidney-tonifying and stasis-removing acupuncture therapy combined with radiofrequency surgery can notoriously enhance the clinical therapy effect and enhance the quality of life of patients, and the detection of NTx, BGP, and CYFRA21-1 indicators can effectively predict the prognosis.

Treating Pb-contaminated clay slurry by three curing agents.

Each year, extensive dredged clay slurries containing heavy metals need to be treated before being reused; in such contaminated slurries, lead (Pb) is frequently identified. Quicklime (CaO)-activated ground granulated blast-furnace slag (GGBS), magnesium (MgO)-activated GGBS, and ordinary Portland cement (OPC) are usually used to remediate the lead (Pb)-contaminated soil; nevertheless, using these curing agents (or binders), particularly CaO-GGBS and MgO-GGBS, to treat Pb-contaminated slurry with high water content is rarely reported. Moreover, inconsistent results were obtained from previous studies in terms of the mechanical and leaching performance of Pb-contaminated soils with the three binders. Based on the above-mentioned reasons, this study used CaO-GGBS, MgO-GGBS, and OPC to treat the Pb-contaminated clay slurry, and compared the effectiveness of the three binders in improving the mechanical and leaching properties of the slurry. Laboratory tests were performed to examine the leaching, strength, mineralogical, and micro-structural performance of treated clay slurries. The results showed that GGBS-based binders were more effective than OPC in improving the strength and Pb leachability of contaminated slurries. When suitable ratios between activators (CaO and MgO) and GGBS were used, a similar or even higher UCS was produced by CaO-GGBS than MgO-GGBS. Similar leachate pH and Pb leachability could be achieved between CaO-GGBS- and MgO-GGBS-treated contaminated clay slurries. Therefore, it is not rigorous to state that MgO-GGBS is better in improving the strength and leachability of Pb-contaminated soils than CaO-GGBS only by comparing the two GGBS-binders based on the same activator/GGBS ratio, as reported in some previous studies. The leachability of Pb was affected by the pH, but the addition of GGBS facilitated the decrease of Pb leachability in slurries. The XRD result showed the formation of CSH and Pb(OH)2, which facilitated the reduction of Pb leachability.

[Molecular mechanism of astragaloside â £ against atherosclerosis by regulating miR-17-5p and PCSK9/VLDLR signal pathway].

This study explores the regulatory effect of astragaloside Ⅳ on miR-17-5 p and its downstream proprotein convertase subtillisin/kexin type 9(PCSK9)/very low density lipoprotein receptor(VLDLR) signal pathway, aiming at elucidating the mechanism of astragaloside Ⅳ against atherosclerosis(AS). In cell experiment, oxidized low-density lipoprotein(ox-LDL) was used for endothelial cell injury modeling with vascular smooth muscle cells(VSMCs). Then cells were classified into the model group, miR-17-5 p inhibitor group, blank serum group, and astragaloside Ⅳ-containing serum group based on the invention. Afterward, cell viability and the expression of miR-17-5 p, VLDLR, and PCSK9 mRNA and protein in cells in each group were detected. In animal experiment, 15 C57 BL/6 mice were used as the control group, and 45 ApoE~(-/-) mice were classified into the model group, miR-17-5 p inhibitor group, and astragaloside Ⅳ group, with 15 mice in each group. After 8 weeks of intervention, the peripheral serum levels of interleukin-6(IL-6), interleukin-10(IL-10), and tumor necrosis factor-α(TNF-α), and the expression of miR-17-5 p, VLDLR, and PCSK9 mRNA in the aorta of mice were detected. The pathological changes of mice in each group were observed. According to the cell experiment, VSMC viability in the miR-17-5 p inhibitor group and the astragaloside Ⅳ-containing serum group was higher than that in the model group(P<0.05). The mRNA and protein expression of miR-17-5 p and VLDLR in VSMCs in the miR-17-5 p inhibitor group and the astragaloside Ⅳ-containing serum group was lower than that in the model group(P<0.05), but the mRNA and protein expression of PCSK9 was higher than that in the model group(P<0.05). As for the animal experiment, the levels of IL-6 and TNF-α in the peripheral serum of the miR-17-5 p inhibitor group and the astragaloside Ⅳ group were lower(P<0.05) and the serum level of IL-10 was higher(P<0.05) than that of the model group. The mRNA expression of miR-17-5 p and VLDLR in the aorta in the miR-17-5 p inhibitor group and the astragaloside Ⅳ group was lower(P<0.05), and PCSK9 mRNA expression was higher(P<0.05) than that in the model group. Pathological observation showed mild AS in the miR-17-5 p inhibitor group and the astragaloside Ⅳ group. In summary, astragaloside Ⅳ can prevent the occurrence and development of AS. The mechanism is that it performs targeted regulation of miR-17-5 p, further affecting the PCSK9/VLDLR signal pathway, inhibiting vascular inflammation, and thus alleviating endothelial cell injury.

Machine Learning Applications in Drug Repurposing.

The coronavirus disease (COVID-19) has led to an rush to repurpose existing drugs, although the underlying evidence base is of variable quality. Drug repurposing is a technique by taking advantage of existing known drugs or drug combinations to be explored in an unexpected medical scenario. Drug repurposing, hence, plays a vital role in accelerating the pre-clinical process of designing novel drugs by saving time and cost compared to the traditional de novo drug discovery processes. Since drug repurposing depends on massive observed data from existing drugs and diseases, the tremendous growth of publicly available large-scale machine learning methods supplies the state-of-the-art application of data science to signaling disease, medicine, therapeutics, and identifying targets with the least error. In this article, we introduce guidelines on strategies and options of utilizing machine learning approaches for accelerating drug repurposing. We discuss how to employ machine learning methods in studying precision medicine, and as an instance, how machine learning approaches can accelerate COVID-19 drug repurposing by developing Chinese traditional medicine therapy. This article provides a strong reasonableness for employing machine learning methods for drug repurposing, including during fighting for COVID-19 pandemic.

Scientific Knowledge Graph of Acupuncture for Migraine: A Bibliometric Analysis from 2000 to 2019.

OBJECTIVE: This study aims to explore the trend and knowledge mapping of acupuncture for migraine through bibliometrics. METHODS: It retrieved the literature on acupuncture for migraine in the Web of Science database from 2000 to 2019, and then resorted to CiteSpace to conduct bibliometric analysis to attain the knowledge mapping. RESULTS: The total number of publications each year has increased year by year, and the average annual growth rate from 2000 to 2009 was 15.57%, while from 2010 to 2019 was 6.35%, with a faster growth rate from 2000 to 2009. According to the cluster analysis of institutions, authors, cited references, and keywords, 10, 7, 12, and 10 categories were gained from 2000 to 2019. The most productive countries, institutions, and authors are the USA and China, Technical University of Munich and Beijing University of Chinese Medicine, Linde K and Liang FR from 2000 to 2019, whose frequency is 119/103, 28/24, and 28/24, respectively. However, the most important of them are Canada, Sichuan University, and Witt CM. Owing to their highest centrality, they are 0.86, 0.54, and 0.27 separately. Moreover, cited references that contributed to the most co-citations are Linde K (2005), yet, the most vital cited reference is Karst M (2001). Keywords such as migraine, acupuncture, headache, pain, and randomized controlled trial are the most frequently used. However, needle acupuncture is the crucial keyword. In the cluster analysis of institutions, authors, cited references, and keywords from 2000 to 2019, the largest cluster categories are #0 migraine prophylaxis, #1 randomized controlled trial, #0 episodic migraine, and #0 topiramate treatment. Then, randomized controlled trials of acupuncture prevention and treatment of migraine are the most important research content in this field. CONCLUSION: Through the bibliometric analysis of the research on acupuncture for migraine in the Web of Science database in the past 20 years, the trends and the Knowledge Graph of the country, institution, author, cited reference, and the keyword are acquired, which have an important guiding significance for quickly and accurately positioning the key information in the field.

Protocatechudehyde improves mitochondrial energy metabolism through the HIF1α/PDK1 signaling pathway to mitigate ischemic stroke-elicited internal capsule injury.

ETHNOPHARMACOLOGICAL RELEVANCE: The internal capsule is vulnerable to ischemia, and mild ischemic stroke often results in lesion of the internal capsule, manifested as contralateral hemiplegia. Protocatechudehyde (PCA), a potential neuroprotective agent, has shown therapeutic effects in the study of a variety of nervous system diseases, including ischemic stroke. AIM OF THE STUDY: The aim of this study was to evaluate the effects of PCA on cerebral ischemia reperfusion (CI/R)-elicited internal capsule injury and to elucidate the role of mitochondrial energy metabolism in the underlying mechanism of neuroprotective effects on ischemic stroke. MATERIALS AND METHODS: A rat tMCAO model was established to investigate the therapeutic effects of intravenous PCA (20, 40, and 80 mg/kg, once per day, continued for 7 days) on CI/R-induced internal capsule injury and the regulation of PCA on molecules related to mitochondrial energy metabolism. In vitro, an OGD/R model of PC12 cells was established to further verify the therapeutic mechanism of PCA. RESULTS: Results showed that PCA dose-dependently attenuated neurological deficit, reduced cerebral infarction, alleviated histopathological damage, and improved mitochondrial ultrastructure of the internal capsule after CI/R. Moreover, PCA reversed the upregulation of HIF1α, PDK1 and pPDHA1 expression induced by CI/R and significantly increased the content of acetyl-CoA, ATP, and the activity of ATP synthase. In vitro, PCA treatment promoted cell survival, inhibited apoptosis, attenuated the dissipation of mitochondrial membrane potential in OGD/R-treated PC12 cells, and these therapeutic effects were reversed by the combination of cobalt chloride (CoCl2), a specific pharmacological inducer of HIF1a expression. CONCLUSIONS: These results indicate that PCA exerts a protective effect against CI/R-induced internal capsule injury and improves mitochondrial energy metabolism in the internal capsule, and the mechanism is associated with the inhibition of HIF1α/PDK1 signaling pathway.

Neural Fuyuan Formula promotes neural plasticity through BDNF/Trkß signaling pathway.

BACKGROUND: Depression after stroke is usually a chronic process, which was associated with many health problems. This study was aimed to investigate the underline mechanism of the effect of Neural Fuyuan Formula (NFF) in post-stroke depression and the role of brain-derived neurotrophic factor (BDNF) in the signal pathway regulated by NFF. METHODS: A rat post-stroke depression model was established. Synaptic plasticity of rat was detected by Electron microscopy. The expression of BDNF signaling proteins and synapse related proteins were measured by Western blot. The expression of Synapsin-1 (SYN1) in rat and the culture neurons was detected by Western blot and immunofluorescence. Dendritic complexity was also measured. RESULTS: NFF could attenuate the synapse change in the post-stroke depression (PSD) model rat. NFF increased the expression of BDNF signaling proteins and synapse related proteins of the PSD model rat (P&0.05). NFF increased the expression of SYN1 in rat and the culture neurons (P&0.05). NFF could also increase the dendritic complexity in culture neurons (P&0.05). CONCLUSIONS: NFF promoted recovery of neurological function through BDNF signaling pathways, which further affirm the curative effect of NFF for treatment of post-stroke depression.

Paeonol administration alleviates cognitive deficits and attenuates neural pathological changes in APP/PS1 mice.

Alzheimer's disease typically presents with impaired cognition and pathological morphologic changes, including the accumulation of amyloid-ß plaques. Disease-modifying drugs are in urgent need as neuroprotective therapies. Exploration of novel therapeutics for alleviating symptoms of Alzheimer's disease has found promise in plant extracts of functional phenols. Paeonol is a water-soluble phenolic substance that has been shown to confer diverse biological effects, including neuroprotection. An Alzheimer's disease model of APP/PS1 double transgenic mice was used in this study, and the therapeutic effects of paeonol were assessed after three weeks' administration. It was found that paeonol treatment significantly increased behavioral performance in the Morris water maze test and increased discrimination rate in the novel object recognition test compared to vehicle-treated APP/PS1 mice. Histologically, paeonol treatment significantly alleviated the Aß plaque burden, reduced neural loss, inhibited microglia activation, and decreased neuroinflammation in the brain of APP/PS1 mice. In addition, a number of Alzheimer's disease-related synaptic plasticity deficits were ameliorated. The present results indicate that paeonol significantly relieved amyloid-ß deposition and amyloid-ß -mediated neuropathology in the brain of APP/PS1 mice, suggesting the potential of paeonol as a preventive and therapeutic agent for Alzheimer's disease.

Carbonating MgO for treatment of manganese- and cadmium-contaminated soils.

Ordinary Portland cement (OPC) and lime are commonly used to treat soils contaminated by heavy metals, such as cadmium (Cd) and manganese (Mn). However, the production of these two binders is not sustainable, consuming high energy and emitting high carbon dioxide (CO2). In this contest, this study proposed a novel and sustainable method of carbonating magnesia (MgO) for treatment of Cd- and Mn-contaminated soils, which can sequester CO2 and immobilize Cd and Mn in the soils. To validate the method, a range of experiments were performed. First, MgO and CO2 were used to treat contaminated soils. Then, the properties of the treated soils were evaluated by unconfined compressive strength test, one stage batch leaching test, X-ray diffraction test, and thermogravimetric analysis. It was found that the carbonation process of MgO-treated soils was decelerated by Mn, but not significantly decelerated by Cd. After carbonation, multiple magnesium carbonates were formed in both contaminated soils, and CdCO3 was formed in the Cd-contaminated soil, while MnCO3 was not confidently determined in the Mn-contaminated soil. Both Cd and Mn negatively affected the strength of carbonated MgO-treated soils; nevertheless, if the concentration of Cd or Mn was not more than 8000 mg/kg, 5% MgO-treated soils after carbonation could meet the strength requirement of higher than 1000 kPa. The treatment decreased the Cd leachability to be below the limit for non-hazardous wastes. The leached concentration of Mn was decreased to be lower than the limit of drinking water.

Vitamin D supplementation prior to in vitro fertilisation in women with polycystic ovary syndrome: a protocol of a multicentre randomised, double-blind, placebo-controlled clinical trial.

INTRODUCTION: Polycystic ovary syndrome (PCOS) is one of the leading causes of female infertility, affecting around 5% of women of childbearing age in China. Vitamin D insufficiency is common in women with PCOS and is associated with lower live birth rates. However, evidence regarding the effectiveness of vitamin D supplementation in women with PCOS is inconclusive. This multicentre randomised, double-blinded, placebo-controlled trial aims to evaluate the effectiveness of vitamin D supplementation prior to in vitro fertilisation (IVF) on the live birth rate in women with PCOS. METHODS AND ANALYSIS: We plan to enrol women with PCOS scheduled for IVF. After informed consent, eligible participants will be randomised in a 1:1 ratio to receive oral capsules of 4000 IU vitamin D per day or placebo for around 12 weeks until the day of triggering. All IVF procedures will be carried out routinely in each centre. The primary outcome is live birth after the first embryo transfer. The primary analysis will be by intention-to-treat analysis. To demonstrate or refute that treatment with vitamin D results in a 10% higher live birth rate than treatment with placebo, we need to recruit 860 women (48% vs 38% difference, anticipating 10% loss to follow-up and non-compliance, significance level 0.05 and power 80%). ETHICS AND DISSEMINATION: This study has been approved by the Ethics Committee in Women's Hospital of Zhejiang University on 2 March 2020 (reference number: IRB-20200035-R). All participants will provide written informed consent before randomisation. The results of the study will be submitted to scientific conferences and a peer-reviewed journal. TRIAL REGISTRATION NUMBER: NCT04082650.

The clinical value of the changes of peripheral lymphocyte subsets absolute counts in patients with non-small cell lung cancer.

INTRODUCTION: Immune function strongly influences the outcome of patients with non-small cell lung cancer (NSCLC). It's vital to understand the immune state of patients through detecting the percentage and number of lymphocyte subsets accurately, and helpful to evaluate conditions of prognosis and adjust treatment for patients. METHODS: We conducted a retrospective cohort study in First Teaching Hospital of Tianjin University of Traditional Chinese Medicine, Tianjin, China. The absolute counts and percentages of CD3+, CD3 + CD4+, CD3 + CD8+, B and NK cells were determined by single platform technologies. 172 patients received treatment including surgery or chemotherapy after surgery. The factors affecting disease progression were analyzed by Binary Logistic regression. Progression free survival (PFS) calculating survivals were with the method of Kaplan-Meier. The log-rank test and cox's proportional hazard regression (enter method) were used for univariable and multivariable analyses respectively. RESULTS: Relative to normal controls, patients with NSCLC at different stages showed decreased absolute lymphocyte count obviously, rather than lymphocyte percentages. Different treatments had unlike influence on the homeostasis of lymphocytes and the effects last for a long time. Logistic regression showed CD3 + CD4+ and CD3 + CD8+ could contribute to favorable prognosis. Multivariate analysis of prognostic factors of PFS showed CD3 + CD4+ cell was independent factor for predicting PFS. CONCLUSIONS: The absolute count of CD3+, CD3 + CD4+, CD3 + CD8+, B and NK cells were better indication of the patient's immune state than percentages of each lymphocyte subsets. Immune function was impaired in patients with non-small cell lung. The high level of baseline absolute CD3 + CD4+ cells count contributed to longer progression free survival. Chinese Clinic Trial Registry number: ChiCTR-IOR-17014139; Registry date: 2017/12/25.

Effects of Combined Bushen Zhichan Recipe and Levodopa in a Rodent Model of Parkinson Disease: Potential Mechanisms.

BACKGROUND Parkinson disease is characterized by the loss of neurons in the substantia nigra, and under pathological conditions, glutamate can produce excitotoxic effects on nerve cells. The astrocytic excitatory amino acid transporter (EAAT) 1 can be functionally upregulated and targeted to functional compartments, resulting in reduced excitotoxicity. levodopa is the gold standard for the treatment of Parkinson disease, but prolonged levodopa treatment often leads to the development of abnormal involuntary movements. Numerous studies suggest the potential beneficial effects of traditional Chinese medicine on Parkinson disease. MATERIAL AND METHODS We validated the efficacy of a Bushen Zhichan recipe combined with levodopa in a rodent Parkinson disease model and explored its possible mechanisms. RESULTS Rats in the combined levodopa and Bushen Zhichan recipe group performed significantly better than the control group in the open field and forelimb function experiments. The number of midbrain dopaminergic neurons in rats in the levodopa and Bushen Zhichan recipe group was greater compared to controls. The levodopa and Bushen Zhichan recipe group exhibited decreased glutamate receptors and increased γ-aminobutyric acid receptors in the striatum. At the same time, EAAT1 was increased and EAAT2 was synchronized with the number of glutamate receptors. CONCLUSIONS Our results indicate that levodopa combined with Bushen Zhichan recipe significantly improves behavior and protects dopaminergic neurons in a rodent Parkinson disease model, and suggest that the mechanism involves the decrease of excitatory amino acid toxicity and the increase in the expression of EAAT1.

[Effects of "Tiaoyi Sanjiao" acupuncture and moxibustion on cancer-induced fatigue and immune function in patients with advanced non-small cell lung cancer].

OBJECTIVE: To observe the therapeutic effect of "Tiaoyi Sanjiao" （regulating and tonifying the triple energizer）acupuncture and moxibustion in the treatment of cancer-related fatigue of advanced non-small cell lung cancer and observe its influence on lymphocyte count. METHODS: The cancer-related fatigue patients with advanced non-small cell lung cancer who met the inclusive criteria were randomly divided into 2 groups, with 77 cases in each group. Patients in the control group were treated with conventional Chinese and Western medicine. In the treatment group, on the base of the treatment as the control group, "Tiaoyi Sanjiao" acupuncture and moxibustion was applied（mild moxibustion at Danzhong ［CV17］, Zhongwan［CV12］, Qihai［CV6］, bilateral Zusanli［ST36］, and acupuncture at bilateral Xuehai［SP10］, Waiguan［SJ5］, Taichong［LR3］）. KPS score, Piper scale, percentage and absolute count of lymphocyte subsets of the two groups before and after treatment were observed. RESULTS: Compared with pre-treatment, the KPS scores of both groups were significantly increased after treatment (P<0.05); the behavioral dimension score of the Piper scale, and the percentage of B cells of the control group decreased significantly(P<0.05); the behavioral dimension, emotional dimension, sensory dimension scores, and total score of the Piper scale in the treatment group were significantly reduced(P<0.05), while the percentage of CD3+T cells and absolute counts of CD3+T cells, CD4+T cells and CD8+T cells of the treatment group were significantly increased (P<0.05). After treatment, in comparison with the control group, behavioral dimension score of the Piper scale in the treatment group decreased significantly(P<0.05); the percentage and absolute counts of B cells and CD4+T cells, the absolute count of CD3+T cells in the treatment group were up-regulated (P<0.05). CONCLUSION: "Tiaoyi Sanjiao" acupuncture and moxibustion can significantly alleviate the fatigue state and improve the quality of life in patients with advanced non-small cell lung cancer, which may be achieved by regulating the number of lymphocytes and improving immune function.