The combination of Syto16/7-AAD allows the detection of apoptosis at an earlier stage than that of annexin V/7-AAD. The effects of the earlier apoptosis of spermatozoa on fertilization rate and post-embryonic development have not been investigated systematically. The objective of this study is to assess the relationship between the earlier apoptosis of spermatozoa in IVF patients and seminal parameters (concentration and morphology), to assess the effect of the earlier apoptosis of spermatozoa on ART outcomes, especially with regard to clinical pregnancy. The earlier apoptosis of spermatozoa was detected by syto16/7AAD using flow cytometry (FCM) in 98 couples (98 IVF) before and after density gradient centrifugation (DGC), and its relationships with the fertilization rate, the normal fertilization rate, the cleavage rate, the good embryo rate and the cumulative clinical pregnancy rate were assessed. Meanwhile, the correlation with seminal parameters (concentration, motility and morphology) was also analyzed. The rate of the earlier apoptosis was significantly negatively correlated to sperm concentration( r = −.302, p = .003) and membrane integrity rate( r = −.862; p < .01) after DGC. There was also a positive correlation between the rate of the earlier apoptosis and sperm mortality rate ( r = .225, p = .027) after DGC. A highly significantly negative correlation was found between the rate of the earlier apoptosis and outcomes in IVF patients: the fertilization rate and the normal fertilization rate decreased as the rate of the earlier apoptosis increased ( r = −.271, −.288; p = .008,.004). The cumulative clinical pregnancy rate also significantly differed between patients with the earlier apoptotic rate ≥15% and <15% (57.89% vs. 81.69%, p < .01) after DGC. The earlier apoptosis of spermatozoa seems to affect fertilization, however, it does not affect the early embryo development proficiency in IVF procedures. Because of the decreased fertilization rate, the earlier apoptosis of spermatozoa can compromise the number of available embryos and good embryos, resulting in cumulative clinical pregnancy loss.

Aging-related cognitive impairment seriously diminishes individuals' life quality. Icariin (ICA), a natural flavonoid separated from the herb Epimedium, is applied in the food industry to bolster immunity and cognitive function in Chinese culture, demonstrating considerable potential in alleviating aging-related cognitive impairment. However, the mechanisms by which ICA mitigates aging-related cognitive impairment have yet to be elucidated. In the study, an 8 week ICA administration strongly improved spatial learning and memory ability, reduced neural damage, and restored hippocampal mitochondrial ultrastructure in mice subjected to d-galactose (d-gal) induction. Mechanically, ICA alleviated colonic pathology and upregulated the expression of tight junction proteins. Moreover, ICA reshaped microbial composition, enriched short-chain fatty acid (SCFA)-producing genera, and upregulated microbiota-derived SCFA contents. Additionally, ICA enhanced cognitively related anti-inflammatory properties and antioxidant capacity. Intriguingly, SCFAs regulated by ICA mitigated mitochondrial dysfunction in vitro, namely, reversing inflammatory cytokine levels and antioxidant capacity, elevating ATP contents, and mitochondrial membrane potential. Furthermore, SCFAs regulated by ICA alleviated mitochondrial dysfunction by enhancing the oxidative phosphorylation pathway and upregulating mRNA expression of genes related to mitochondrial respiratory chain, thus improving cognitive function. The findings suggest that ICA alleviates d-gal-induced cognitive impairment via modulation of the gut-brain axis and mitochondrial function. The investigation underscores the potential therapeutic benefits of incorporating an ICA-enriched diet for cognitive enhancement.

BACKGROUND:With the acceleration of population ageing, there has been increasing attention to ageing-related diseases, especially mild cognitive impairment (MCI). Vitamin C (VC), known as ascorbic acid, has demonstrated potential anti-ageing effects and may offer protection against MCI. The study sought to investigate the underlying mechanisms by which VC protects against MCI induced by D-galactose (D-gal). APPROACHES:ICR mice were subjected to D-gal to elicit MCI and were subsequently administered VC for 8 weeks. The therapeutic effects of VC were assessed by behavioural evaluations and hippocampal pathology. To investigate the potential mechanisms, 16S rDNA sequencing, gas chromatography, and Spearman correlation analysis were employed. RESULTS:VC supplementation significantly improved spatial learning and memory functions while mitigating hippocampal neuronal damage in D-gal-induced mice. Additionally, VC enhanced cognitive-related anti-inflammatory properties and antioxidant capacity. VC markedly mitigated the colonic pathological damage. Notably, VC led to increased microbial diversity, particularly the enrichment of genera that produce short-chain fatty acids (SCFAs), such as Akkermansia, Ruminococcus, and Butyricicoccus. Interestingly, levels of SCFAs (acetic acid, propionic acid, and butyric acid, etc.) were elevated following VC administration. The Spearman correlation analysis revealed that SCFAs levels were positively correlated with the abundance of the probiotics (Ruminococcus and Butyricicoccus) in response to VC. CONCLUSIONS:VC supplementation may mitigate MCI, potentially through modulation of intestinal flora and SCFAs production. These results establish a foundation for the application of VC in the management of MCI and underscore its potential as a therapeutic strategy for ageing-related diseases.

An entry from the Cambridge Structural Database, the world’s repository for small molecule crystal structures. The entry contains experimental data from a crystal diffraction study. The deposited dataset for this entry is freely available from the CCDC and typically includes 3D coordinates, cell parameters, space group, experimental conditions and quality measures.

Chimeric antigen receptor T (CAR-T) cells are genetically modified T cells with redirected specificity and potent T-cell-mediated cytotoxicity toward malignant cells. Despite several CAR-T products being approved and commercialized in the USA, Europe, and China, CAR-T products still require additional optimization to ensure reproducible and cost-effective manufacture. Here, we investigated the critical parameters in the CD3+ T-cell isolation process that significantly impacted CAR-T manufacturing's success. CAR-T cells were prepared from cryopreserved peripheral blood mononuclear cells (PBMC). The thawed PBMC was rested overnight before the CD3+ T cell isolation process using CTS™ Dynabeads™ CD3/CD28. Different isolation media, cell-bead co-incubation time, and cell density were examined in this study. Activated CD3+ T cells were transduced with a gamma retroviral vector carrying the CD19 or BCMA CAR sequence. The CAR-T cells proliferated in a culture medium supplemented with interleukin 2 (IL-2). CD14+ monocytes hindered T-cell isolation when X-VIVO 15 basic medium was used as the selection buffer. The activation of T cells was blocked because monocytes actively engulfed CD3/28 beads. In contrast, when DPBS was the selection medium, the T-cell isolation and activation were no longer blocked, even in patients whose PBMC contained abnormally high CD14+ monocytes and a low level of CD3+ T cells. In this study, we discovered that selecting CD3+ T-cell isolation media is critical for improving T-cell activation, transduction, and CAR-T proliferation. Using DPBS as a CD3+ T cell isolation buffer significantly improved the success rate and shortened the duration of CAR-T production. The optimized process has been successfully applied in our ongoing clinical trials. Trial registration NCT03798509: Human CD19 Targeted T Cells Injection Therapy for Relapsed and Refractory CD19-positive Leukemia. Date of registration: January 10, 2019. NCT03720457: Human CD19 Targeted T Cells Injection (CD19 CAR-T) Therapy for Relapsed and Refractory CD19-positive Lymphoma. Date of registration: October 25, 2018. NCT04003168: Human BCMA Targeted T Cells Injection Therapy for BCMA-positive Relapsed/Refractory Multiple Myeloma. Date of registration: July 1, 2019

Chemical investigation of Dendrobium plicatile Lindl resulted in the isolation and identification of one new bibenzyl, 2-chloro-3, 4'-dihydroxy-3',5-dimethoxybibenzyl (1), as well as 15 known stilbenoids. The structures of this new compound was elucidated by extensive spectroscopic analysis, including HRESIMS, 1H and 13C NMR, DEPT, HMBC, COSY, HMQC, NOESY. Compounds 2, 3 and 5 were obtained from this genus for the first time, compounds 8, 10, 13 and 14 were obtained from this plant for the first time. In addition, the new compound exhibited potent cytotoxic activities against the human breast cancer (MDA-MB231) cell line, the hepatocellular carcinoma (HepG2) cell line and the human lung carcinoma (A549) cell line, with IC50 3.41, 3.02, 2.80 μM, respectively.

In this paper, two natural products: chelerythrine (Che) and sanguinarine (San) were systematically explored for aggregation-induced emission enhancement (AIEE) characteristics by a series of spectroscopic and theoretical experiments.

Two new stilbenoids, named 2,3 -dimethoxyl-7-hydroxyl-1,4-phenanthrenedione (1) and 2-methoxyl-3-methyl-7-hydroxyl-9,10-dihydro-1,4-phenanthrenedione (2), together with two known stilbenoids including densiflorol B (3) and ephemeranthoquinone (4), were isolated from aerial parts of Flickingeria fimbriata (Bl.) Hawkes. The structures of two new compounds were elucidated by extensive spectroscopic analysis, including HRESIMS, 1H and 13C NMR, DEPT, HMBC, COSY, HMQC, NOESY. All the compounds were obtained from this genus for the first time. In addition, they all exhibited moderate cytotoxic activities against HepG2 cell lines.

Chemical investigation of Flickingeria fimbriata (Bl.) Hawkes (Orchidaceae) resulted in the isolation and identification of one new dihydrophenanthrene, 1,2,5,6,7-pentamethoxy-9,10-dihydrophenanthrene (1), together with seven known dihydrophenanthrenes, erianthridin (2), coelonin (3), 4-methoxy-9,10-dihydrophenanthrene-1,2,7-triol (4), lusianthridin (5), ephemeranthol A (6), flavanthridin (7) and hircinol (8), four known phenanthrenes, epheranthol B (9), nudol (10), denthyrsinin (11) and confusarin (12), and one known bibenzyl, batatasin III (13). The structure of the new compound was elucidated by spectroscopic analysis (HRMS, 1D and 2D NMR). All the compounds were isolated from F. fimbriata for the first time except for compounds 5 and 12, and compounds 1, 3, 4, 8, 10, 11 and 13 were obtained from this genus for the first time. Compounds 1-4 showed moderate cytotoxic activity against HepG2 cells.

Human Nestin (hNestin) has been found to express in melanoma, and its expression is positively correlated with the advanced stage of melanoma. However, the precise role of hNestin in the development of melanoma has not been fully understood. The present study aimed to explore the role of hNestin in the proliferation and invasion of melanoma cells. The lentivirus vector carrying a short hairpin RNAs (shRNAs) targeting hNestin (hNestin-shRNA-LV) was stably infected into human melanoma cells UACC903, which expressed high levels of hNestin. The effects of hNestin knockdown on the proliferation, apoptosis, migration of melanoma cells and the related signaling pathways were investigated by immunofluorence, Western blotting and reverse transcription polymerase chain reaction (RT-PCR), respectively. The results showed that hNestin was expressed in most melanoma specimens and the melanoma cells studied. Knockdown of hNestin expression significantly inhibited the proliferation of melanoma cells, blocked the formation of cell colony, arrested cell cycle at G1/S stage and suppressed the activation of Akt and GSK3β. hNestin-silent cells also showed a sheet-like appearance with tight cell-cell adhesion, decreased membrane expression of N-cadherin and β-catenin, and attenuated migration. Furthermore, hNestin silence resulted in the inhibition of tumor growth in vivo. Our study indicates that hNestin knockdown suppresses the proliferation of melanoma cells, which might be through affecting Akt-GSK3β-Rb pathway-mediated G1/S arrest, and hNestin silence inhibits the migration by selectively modulating the expression of cell adhesion molecules in the process of epithelial-mesenchymal transition.

Five kinds of berberine derivatives with hydroxy, methyl, ethyl, benzyl and (4-methyl)benzyl group at 13 position have been synthesized and characterized. 13-hydroxyberberine chloride exhibits aggregation induced emission enhancement (AIEE) property because of hydroxy group, which is beneficial to increase the rigidity of molecule by enlarging conjugated system. Optical properties in pure solution, CH3OH/H2O mixed solution, amorphous and crystalline state were comparatively investigated. Polymeric morphology and particle size of 13-hydroxyberberine chloride with different water fractions (090 vol%) were obtained by scanning electron microscope (SEM) and dynamic light scattering (DLS) method respectively, which provided reasonable explanation that the formation of small globular nanoparticles in mixed solution is conducive to the fluorescence emission. The single crystal structure of 13-hydroxyberberine chloride was determined by single-crystal X-ray diffraction. Crystallographic data indicated that the main mechanism of the AIEE phenomenon is the existences of J-aggregation (head to tail dipole stacking) combined with molecular planarization. The calculation done by DFT showed that the HOMO-LUMO bandgap is in accordance with experimental data. To further explore the biomedical application of 13-hydroxyberberine, its cell viability and cell imaging performance were examined, which demonstrate that 13-hydroxyberberine shows definite fluorescent intensity. In all, 13-hydroxyberberine should be a promising candidate for different biomedical application such as pH fluorescence probe because of its response to the stimuli of pH value. (C) 2017 Elsevier B.V. All rights reserved.

Laryngeal cancer is the most common head and neck cancer (skin excluded) with the increasing rates of morbidity and mortality in the world. The emerging roles of microRNAs (miRs) in laryngeal cancer have been deeply investigated in recent years. Deregulated miRs are frequently detected in tissues and cells of laryngeal cancer, which work as oncogenes or tumor supressors to regulate cancer cell proliferation, metastasis and invasion, etc. Here we reviewed the recognized roles of miRs in the diagnosis, prognosis and therapy of laryngeal cancer. Although there are lots of challenges in miRs including sensitivity, specificity, accuracy and safety, the growing improvements of miRs in laryngeal cancer remain encouraging and promising.

Increasing evidence shows that oxidative stress and the hyperphosphorylation of tau protein play essential roles in the progression of Alzheimer's disease (AD). Quercetin is a major flavonoid that has anti-oxidant, anti-cancer and anti-inflammatory properties. We investigated the neuroprotective effects of quercetin to HT22 cells (a cell line from mouse hippocampal neurons). We found that Okadaic acid (OA) induced the hyperphosphorylation of tau protein at Ser199, Ser396, Thr205, and Thr231 and produced oxidative stress to the HT22 cells. The oxidative stress suppressed the cell viability and decreased the levels of lactate dehydrogenase (LDH), superoxide dismutase (SOD), mitochondria membrane potential (MMP) and Glutathione peroxidase (GSH-Px). It up-regulated malondialdehyde (MDA) production and intracellular reactive oxygen species (ROS). In addition, phosphoinositide 3 kinase/protein kinase B/Glycogen synthase kinase3β (PI3K/Akt/GSK3β) and mitogen activated protein kinase (MAPK) were also involved in this process. We found that pre-treatment with quercetin can inhibited OA-induced the hyperphosphorylation of tau protein and oxidative stress. Moreover, pre-treatment with quercetin not only inhibited OA-induced apoptosis via the reduction of Bax, and up-regulation of cleaved caspase 3, but also via the inhibition of PI3K/Akt/GSK3β, MAPKs and activation of NF-κB p65. Our findings suggest the therapeutic potential of quercetin to treat AD.

Nestin is a neuroepithelial stem cell marker that is expressed in some types of tumor cells. Recent reports suggest that Nestin may be closely related to malignant cell proliferation and migration. Acute leukemia (AL) is characterized by a lack of differentiation, which results in uncontrolled proliferation in the bone marrow and accumulation of immature cells. The expression and function of Nestin in AL is unclear. We investigated Nestin immunohistochemical patterns of 87 patients that included 47 cases of acute myeloid leukemia (AML) and 40 cases of acute lymphoblastic leukemia (ALL), and 20 patients in complete remission (CR) from AML or ALL. We also investigated the clinico-pathological features of 87 cases of AL and their CR and overall survival (OS). Nestin was expressed in leukemic blasts and mature granulocytic cells in most cases (39/47) of AML. Conversely, Nestin was expressed in mature granulocytic cells in fewer cases (6/40) of ALL, but not in blasts. Nestin expression appeared in leukemic blasts of AML, but not ALL. Nestin expression in AML blast cells was not associated with CR or OS. We provide evidence that Nestin is expressed in AL and might be a useful immunohistochemical marker for identifying AML and ALL.

Alzheimer's disease (AD) is a neurodegenerative disease characterized by deposit of amyloid plaques and neurofibrillary tangles and oxidative stress plays an essential role in the pathogenesis of AD. Jatrorrhizine (JAT), a Coptidis Rhizome, has multiple biological functions such as anti-oxidation and anti-inflammation. Herein, we investigated the neuroprotective effects of JAT on okadaic acid (OA)- induced cytotoxicity and apoptosis in HT22 cells. Following the exposure to 80 nmol/L OA for 12h, the reduction in cell survival, activities of superoxide dismutase, glutathione peroxidase and mitochondria membrane potential has been shown in HT22 cells. In contrast, OA increased levels of lactate dehydrogenase, malondialdehyde production and intracellular reactive oxygen species. OA also enhanced the expression of Bax but decreased the levels of Bcl-2, OA also upregulated the expression of cleaved caspase-3, phosphorylated extracellular signal-regulated kinases 1/2, phosphorylated c-Jun N-terminal kinases, phosphorylated p38 and NF-kappa B p65 subunit in HT22 cells and this up-regulation was attenuated by JAT which was pre-incubated for 12h in the cells prior to OA exposure. In conclusion, our data present the protective role of JAT in OA induced cytotoxicity, via its antioxidant and anti-apoptotic properties by inhibiting the mitogen-activated protein kinases pathways in HT22 hippocampal neurons. These results indicate that JAT may be the potential target to treat AD induced by oxidative stress and apoptosis.

Objective To investigate the level of serum anti-Mullerian hormone( AMH) in patients with pelvic inflammatory disease( PID) and normal population,and explore the expression pattern and clinical significance of AMH in PID patients. Methods A total of 155 PID patients and 67 healthy women participated in this study. All the subjects were divided into five groups according to age,such as group A including less than 25 years old,group B including 25 to 30 years old,group C including 31 to 35 years old,group D including 36 to 40 years old,and group E including more than 40 years old. Serum AMH level was measured by ELISA and analyzed by SPSS 16.0. Results The average AMH level was( 4.48± 0.36) ng / m L in patients with mild and moderate PID,and was negatively correlated with age( r =-0. 432,P 0. 01).There was a statistical difference between groups A,B and groups C,D and E( P 0. 05),and between group E and groups A,B and C( P 0.01). The average AMH level was( 2.12± 0.26) ng / m L in patients with severe PID,and was negatively correlated with age( r =-0.428,P 0.05). The level of serum AMH markedly dropped in more than 40 years old women. There was a statistical difference between group E and other groups( P 0.01). The average AMH level was( 3.91± 0.51) ng / m L in normal women,and the level of serum AMH was negatively correlated with age( r =-0.742,P 0.01). There was a statistical difference between group E and other groups( P 0.01). There was a statistical difference of the average AMH level between serious PID patients and moderate PID patients as well as normal women based on the same age( P0.05). Conclusion Age is an important factor affecting the level of AMH in normal women and PID patients. There is a significant difference of the serum AMH levels between PID patients and normal women based on the same age.

Recent studies demonstrated that the heart of 1‐day‐old neonatal mice could regenerate, with Wt1 + EPDC s migrating into myocardial regions after partial surgical resection, but this capacity was lost by 7 days of age. By treatment with Tβ4 to maintain Wt1 expression and retain the migrating feature of EPDC s in neonatal mice, we explored the possibility of restoring the cardiac regeneration potential of mice. We intraperitoneally injected Tβ4 into 1‐day‐old mice on daily basis and then apical resection was performed on the mice 7 days later. Twenty one days after the resection, morphological analysis revealed that the Tβ4‐treated mice regenerated the resected ventricular apex, while the mice in PBS control group developed significant fibrosis without apical regeneration. The Tβ4‐treated mice had significantly better ventricular ejection fraction and fractional shortening than controls. During the process of regeneration, Wt1 + EPDC s migrated into myocardial region and some of them expressed Islet1 and the markers for mature cardiomyocytes, such as cTnT and SαA. These characteristics of Wt1 + EPDC s were also seen in the heart regeneration of mice subjected to apical resection 1 day after birth. Tβ4 has no essential effect on cell cycle activity as no disruption of actin filaments was observed in Tβ4‐treated hearts. These results revealed that the cardiac regeneration potential of neonatal mice could be extended to the 7th post‐natal day by Tβ4 and Wt1 + EPDC s mobilization might play an important role in the extension.

阿尔茨海默病（Alzheimer′s disease ，AD ）是一种以认知能力下降为特征的渐进性脑退行性疾病，发病与年龄的增长、人体的衰老密不可分。细胞组织学层面可表现为神经元凋亡以及神经突触的退化。病理学主要表现为以β类淀粉样蛋白（Aβ）沉积为基础的老年斑（SPs），以及过度磷酸化 Tau蛋白聚集形成的细胞内神经元纤维缠结（NFTs）。