Transcriptomic profiling of human granulosa cells between women with advanced maternal age with different ovarian reserve.

BACKGROUND: Age-related diminished ovarian reserve (DOR) is not absolute. Some advanced maternal age (AMA) still have normal ovarian reserve (NOR) and often show better pregnancy outcomes. Exploring the transcriptomic profile of granulosa cells (GCs) in AMA could lead to new ideas for mitigating age-related diminished ovarian reserve. AIM: This study aimed to analyze the transcriptomic profile of GCs in AMA with different ovarian reserve. RESULTS: In total, 6273 statistically significant differential expression genes (DEGs) (|log2fc|> 1, q < 0.05) were screened from the two groups, among which 3436 genes were upregulated, and 2837 genes were downregulated in the DOR group. Through Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis, the potential functions of dysregulated genes in AMA with DOR or NOR were predicted. The GO enrichment analysis revealed that the DEGs were mainly enriched in obsolete oxidation-reduction process, mitochondrion, metal ion binding, ATP binding, etc. The KEGG pathway enrichment analysis revealed that the above-mentioned DEGs were mainly enriched in ferroptosis, regulation of actin cytoskeleton, oxidative phosphorylation, etc. Meanwhile, verification of the mRNA expression levels of DEGs revealed the possible involvement of "ferroptosis" in age-related diminished ovarian reserve. CONCLUSIONS: From a new clinical perspective, we presented the first data showing the transcriptomic profile in GCs between AMA with different ovarian reserve. At the same time, we identified the role of ferroptosis in the GCs of AMA, providing a new biological basis for studying ovarian aging and improving pregnancy outcomes of AMA.

[Effects of acupuncture on hemorrhagic transformation and motor function in stroke patients after intravenous thrombolysis with rt-PA: a prospective cohort study].

OBJECTIVE: To observe the effects of the Xingnao Kaiqiao (regaining consciousness and opening orifices) acupuncture on hemorrhagic transformation and limb motor function after intravenous thrombolysis with recombinant tissue plasminogen activator (rt-PA) in stroke patients. METHODS: A total of 130 stroke patients after rt-PA thrombolytic were divided into an acupuncture group (58 cases, 1 case dropped off) and a non-acupuncture group (72 cases, 7 cases dropped off) according to whether they received acupuncture treatment. Propensity score matching (PSM) was used to match each group, with 38 patients in each group. The patients in the non-acupuncture group received rt-PA thrombolytic therapy and western medical basic treatment. In addition to the basic treatment, the patients in the acupuncture group received Xingnao Kaiqiao acupuncture at Shuigou (GV 26), bilateral Neiguan (PC 6), and ipsilateral Sanyinjiao (SP 6), Chize (LU 5), once a day for 14 days. The incidence of hemorrhagic transformation within 30 days after onset was compared between the two groups. The Fugl-Meyer assessment (FMA) score and activities of daily living (ADL) score were observed at baseline and 30 days, 6 months, 1 year after onset in the two groups. The disability rate at 6 months and 1 year after onset was recorded, and safety was evaluated in both groups. RESULTS: The incidence of hemorrhagic transformation in the acupuncture group was 5.3% (2/38), which was lower than 21.1% (8/38) in the non-acupuncture group (P<0.05). At 30 days, 6 month, and 1 year after onset, the FMA and ADL scores of both groups were higher than those at baseline (P<0.01), and the scores in the acupuncture group were higher than those in the non-acupuncture group (P<0.01). The disability rate in the acupuncture group at 1 year after onset was 10.5% (4/38), which was lower than 28.9% (11/38) in the non-acupuncture group (P<0.05). There was no significant difference in the incidence of adverse events between the two groups (P>0.05). CONCLUSION: The Xingnao Kaiqiao acupuncture method could reduce the incidence of hemorrhagic transformation in stroke patients after intravenous thrombolysis with rt-PA, improve their motor function and daily living ability, and reduce the long-term disability rate.

[Effects of Long-term Sod Cultivation on Chinese Hickory Plantation Soil Fungal Community and Enzyme Activities].

A long-term field experiment was conducted at a Chinese hickory (Carya cathayensis) plantation from 2011 to 2021, with the purpose of researching the effects of long-term sod cultivation on hickory plantation soil fungal communities and enzyme activities and providing experience for ecological management in other plantations. Sod cultivation included oilseed rape (Brassica chinensis, BR), Chinese milk vetch (Astragalus sinicus, AS), and oilseed rape+Chinese milk vetch (BA), and clear tillage (CT) served as a contrast. The soil fertility, fungal community composition and diversity, and soil enzyme activities were determined. The results showed that:① long-term sod cultivation significantly increased soil nutrient contents and availability, and pH increased variably from different sod cultivation treatments (P<0.05). ②The soil fungal community composition was changed by long-term sod cultivation. The relative abundance of Ascomycota, which utilized the readily decomposed organic matter, was increased, whereas the relative abundance of Basidiomycota, which degraded stubborn organic matter, decreased. Long-term sod cultivation shifted the soil dominant genera, as BR and BA increased the relative abundance of somemycorrhizal fungi that could form mutually beneficial structures with dominant plant genera after sod cultivation,whereas AS increased the relative abundance of saprophytic fungi that could decompose the remains of dead plants and animals. The soil fertility factors including pH, available nitrogen, microbial biomass nitrogen, and water-soluble organic carbon were revealed to have a significant influence on the soil fungal composition (P<0.05). ③ Moreover, long-term sod cultivation stimulated the activities of soil enzymes involved in the carbon and nitrogen cycle. Apart from BA, sod cultivation treatments decreased the activities of alkaline phosphatase, which was involved in the soil P turnover. The correlation analysis demonstrated that the correlations between activities of enzymes decomposing carbon and nitrogen and soil fertility were significant (P<0.05 or P<0.01). The activities of phosphatase were positively correlated with soil microbial biomass carbon and nitrogen. Long-term sod cultivation could improve soil nutrient content and availability, optimized soil fungal community structure, and promoted soil nutrient turnover enzyme activities.

Sodium selenite may be not the optimal speciation as an effective therapy for arsenic-induced anxiety-/depression-like behavior.

Major depressive disorder is a serious and prevalent neuropsychiatric disorder, affecting more than 350 million people worldwide. Here, sodium selenite (SS) was selected as the selenite supplement to improve the behavior in a mouse model of depression induced by As. SS may be not the optimal speciation for selenite supplementation and the source of the SS used in the study was not disclosed. There are many mouse models of depression and anxiety; however, in the current study, a classical mouse model of depression was not used. Thus, several questions still need to be further discussed. Taken together, the results indicate that SS may be not the optimal speciation as an effective therapy for As-induced anxiety-/depression-like behavior.

Neuroprotective Effects of the Pannexin-1 Channel Inhibitor: Probenecid on Spinal Cord Injury in Rats.

Proinflammatory immune cell subsets constitute the majority in the local microenvironment after spinal cord injury (SCI), leading to secondary pathological injury. Previous studies have demonstrated that inflammasomes act as an important part of the inflammatory process after SCI. Probenecid, an inhibitor of the Pannexin-1 channel, can inhibit the activation of inflammasomes. This article focuses on the effects of probenecid on the local immune microenvironment, histopathology, and behavior of SCI. Our data show that probenecid inhibited the expression and activation of nucleotide-binding oligomerization domain receptor pyrindomain-containing 1 (NLRP1), apoptosis-associated speck-like protein containing a CARD (ASC) and caspase-1, interleukin-1ß (IL-1ß), and caspase-3 proteins associated with inflammasomes, thereby suppressing the proportion of M1 cells. And consequently, probenecid reduced the lesion area and demyelination in SCI. Moreover, the drug increased the survival of motor neurons, which resulted in tissue repair and improved locomotor function in the injured SC. Altogether, existing studies indicated that probenecid can alleviate inflammation by blocking Pannexin-1 channels to inhibit the expression of caspase-1 and IL-1ß, which in turn restores the balance of immune cell subsets and exerts neuroprotective effects in rats with SCI.

Prevalence and Genetic Analysis of ß-Thalassemia in the Dali Bai Autonomous Prefecture of the Yunnan Province, China.

Background: ß-Thalassemia is the most common monogenetic hemolytic hemoglobin-associated disease in the south of China; the distribution of genetic mutations associated with this condition varies according to geographic regions. This study investigated the prevalence and distribution of ß-thalassemia-associated mutations across different ethnic groups in the Dali Bai Autonomous Prefecture of the Yunnan Province, China. Methods: This cross-sectional study included 4723 participants (15-45 years old) who volunteered for thalassaemia screening from the Dali Bai Autonomous Prefecture from May 2017 to October 2020. Cellulose acetate membrane electrophoresis was used to screen for ß-thalassemia carriers. Genotypic analyses was performed using polymerase chain reaction-based reverse dot blotting and DNA sequencing. Results: The overall prevalence of ß-thalassemia in the study population was 2.01%. The genotypic analyses showed the presence of four types of mutations in the ß-globin gene: CD26 (GAG→AAG), CD56 (GGC→GAC), IVS-II-81 (C→T), and CD121 (GAA→CAA). In contrast to previous studies from other regions of Yunnan Province, our results showed that the prevalence of CD26 mutations was significantly higher than that of the other mutations. Conclusion: Our data suggests that the Dali Autonomous Prefecture is an area with a high prevalence of ß-thalassemia. Moreover, CD26 was the only ß-thalassemia mutation that we have detected. Moreover, the vast majority of the ß-thalassemia mutations observed were CD26.

Exploring the inclusion interaction of estradiol with ß-CD and HP-ß-CD with the help of molecular dynamics simulation as well as multi-spectroscopic approaches.

The inclusion behaviors of estradiol with ß-CD and HP-ß-CD were characterized using molecular dynamics simulation combined with multi-spectroscopic approaches. The findings revealed that estradiol enclosed into the cavity of ß-CD and HP-ß-CD and produced the estradiol-ß-CD and estradiol-HP-ß-CD complexes with the stoichiometry of 1:1. The association constants of the estradiol-ß-CD and estradiol-HP-ß-CD complexes were 3.14 × 104 and 3.22 × 104 M-1 at 298 K, respectively, which declined with rising temperature. The analysis results of thermodynamic parameters confirmed that the dominate interaction forces were the hydrophobic and hydrogen-bonding interactions for stabilizing the estradiol-ß-CD complex, and were the hydrogen bonding interaction and van der Waals forces for stabilizing the estradiol-HP-ß-CD complex. Moreover, it was confirmed from the results of molecular modeling that estradiol inserted into the hydrophobic cavity of ß-CD and HP-ß-CD and form a stable estradiol-CD complexes. And, it is also observed that the phenyl moiety in estradiol is almost parallel to the central axis of ß-CD and HP-ß-CD, and the phenyl moiety was located on wider rim of ß-CD and HP-ß-CD.

Is Human Chorionic Gonadotropin Trigger Beneficial for Natural Cycle Frozen-Thawed Embryo Transfer?

Objective: The aim of this study is to investigate, in ovulatory patients, whether there is a difference in reproductive outcomes following frozen-thawed embryo transfer (FET) in natural cycles (NC) compared to modified natural cycles (mNC). Methods: This retrospective cohort study, performed at the public tertiary fertility clinic, involved all infertile patients undergoing endometrial preparation prior to FET in NC and mNC from January, 2017 to November, 2020. One thousand hundred and sixty-two patients were divided into two groups: mNC group (n = 248) had FET in a NC after ovulation triggering with human chorionic gonadotropin (hCG); NC group (n = 914) had FET in a NC after spontaneous ovulation were observed. The primary outcome was live birth rate. All pregnancy outcomes were analyzed by propensity score matching (PSM) and multivariable logistic regression analyses. Results: The NC group showed a higher live birth rate [344/914 (37.6%) vs. 68/248 (27.4%), P = 0.003; 87/240 (36.3%) vs. 66/240 (27.5%), P = 0.040] than the mNC group before and after PSM analysis. Multivariable analysis also showed mNC to be associated with a decreased likelihood of live birth compared with NC [odds ratio (OR) 95% confidence interval (CI) 0.71 (0.51-0.98), P = 0.039]. Conclusion: For women with regular menstrual cycles, NC-FET may have a higher chance of live birth than that in the mNC-FET cycles. As a consequence, it's critical to avoid hCG triggering as much as possible when FETs utilize a natural cycle strategy for endometrial preparation. Nevertheless, further more well-designed randomized clinical trials are still needed to determine this finding.

Methylene blue ameliorates brain edema in rats with experimental ischemic stroke via inhibiting aquaporin 4 expression.

Brain edema is a common and serious complication of ischemic stroke with limited effective treatment. We previously reported that methylene blue (MB) attenuated ischemic brain edema in rats, but the underlying mechanisms remained unknown. Aquaporin 4 (AQP4) in astrocytes plays a key role in brain edema. We also found that extracellular signal-regulated kinase 1/2 (ERK1/2) activation was involved in the regulation of AQP4 expression in astrocytes. In the present study, we investigated whether AQP4 and ERK1/2 were involved in the protective effect of MB against cerebral edema. Rats were subjected to transient middle cerebral artery occlusion (tMCAO), MB (3 mg/kg, for 30 min) was infused intravenously through the tail vein started immediately after reperfusion and again at 3 h after ischemia (1.5 mg/kg, for 15 min). Brain edema was determined by MRI at 0.5, 2.5, and 48 h after tMCAO. The decreases of apparent diffusion coefficient (ADC) values on diffusion-weighted MRI indicated cytotoxic brain edema, whereas the increase of T2 MRI values reflected vasogenic brain edema. We found that MB infusion significantly ameliorated cytotoxic brain edema at 2.5 and 48 h after tMCAO and decreased vasogenic brain edema at 48 h after tMCAO. In addition, MB infusion blocked the AQP4 increases and ERK1/2 activation in the cerebral cortex in ischemic penumbra at 48 h after tMCAO. In a cell swelling model established in cultured rat astrocyte exposed to glutamate (1 mM), we consistently found that MB (10 µM) attenuated cell swelling, AQP4 increases and ERK1/2 activation. Moreover, the ERK1/2 inhibitor U0126 (10 µM) had the similar effects as MB. These results demonstrate that MB improves brain edema and astrocyte swelling, which may be mediated by the inhibition of AQP4 expression via ERK1/2 pathway, suggesting that MB may be a potential choice for the treatment of brain edema.

[Temporal and Spatial Occurrence of NSAIDs in Taihu Lake and Relevant Risk Assessment].

Non-steroidal anti-inflammatory drugs (NSAIDs) are a class of drug residues with a high frequency of detection in Taihu Lake. However, little information is available about the occurrence of typical NSAID mixtures in Taihu Lake as a whole across the four seasons. Therefore, for each season, the concentrations of five typical NSAIDs including diclofenac, ibuprofen, indomethacin, naproxen, and ketoprofen were monitored in the water of Taihu Lake by the method of high-performance liquid chromatography-mass spectrometry (HPLC-MS/MS) at 19 transects covering the entire lake. The temporal and spatial occurrence of NSAID mixtures in the water of Taihu Lake and their correlation with environmental factors were analyzed, and the mixture risk quotient (MRQ) model was also used to assess the ecological risk of the NSAID mixtures. The research results are as follows:① The concentrations of NSAIDs in the northern, western, and eastern waters of Taihu Lake are at a higher level compared to those in the central waters. Ketoprofen is the main contributor to the contamination of NSAID mixtures in all regions of Taihu Lake. ② The concentrations of NSAIDs in Taihu Lake are higher in summer (15.9-134.3 ng·L-1) and autumn (16.4-144.6 ng·L-1) but lower in spring (25.3-72.5 ng·L-1) and winter (14.6-57.4 ng·L-1), being significantly correlated with water conductivity and pH, respectively. ③ The MRQ model evaluation reveals that there are nine sections in Taihu Lake showing a high ecological risk (MRQ>1) from NSAID mixtures throughout the year. The ecological risk of the NSAID mixtures at a medium or high level (MRQ>0.1) lasts for a long time spanning the spring, summer, and autumn seasons, of which the risk is greatest in autumn. Overall, the pollution caused by the NSAID mixtures in the water of Taihu Lake should not be ignored, and especially great attention should be paid to the pollution in autumn.

Substrate Specificity of Acyltransferase Domains for Efficient Transfer of Acyl Groups.

Acyltransferase domains (ATs) of polyketide synthases (PKSs) are critical for loading of acyl groups on acyl carrier protein domains (A) via self- and trans-acylation reactions, to produce structurally diverse polyketides. However, the interaction specificity between ATs and unusual acyl units is rarely documented. In Streptomycestsukubaensis YN06, we found that AT4FkbB [an AT in the fourth module of tacrolimus (FK506) PKS] transferred both allylmalonyl (allmal) and emthylmalonyl (ethmal) units to ACPs, which was supposed responsible for the production of both FK506 and its analog FK520, respectively. Mutations of five residues in AT4FkbB (Q119A, L185I-V186D-V187T, and F203L) caused decreased efficiency of allmal transfer, but a higher ratio of ethmal transfer, supposedly due to less nucleophilic attacks between Ser599 in the active site of AT4FkbB and the carbonyl carbon in the allmal unit, as observed from molecular dynamics simulations. Furthermore, reverse mutations of these five residues in ethmal-specific ATs to the corresponding residues of AT4FkbB increased its binding affinity to allmal-CoA. Among these residues, Val187 of AT4FkbB mainly contributed to allmal recognition, and V187K mutant produced less FK520 than wild type. Our findings thus suggested that five critical residues within AT4FkbB were important for AT functionality in polyketide extension and potentially for targeting biosynthesis by generating desirable products and eliminating undesirable analogs.

Glutamate Impairs Mitochondria Aerobic Respiration Capacity and Enhances Glycolysis in Cultured Rat Astrocytes.

OBJECTIVE: To study the effect of glutamate on metabolism, shifts in glycolysis and lactate release in rat astrocytes. METHODS: After 10 days, secondary cultured astrocytes were treated with 1 mmol/L glutamate for 1 h, and the oxygen consumption rates (OCR) and extra cellular acidification rate (ECAR) was analyzed using a Seahorse XF 24 Extracellular Flux Analyzer. Cell viability was then evaluated by MTT assay. Moreover, changes in extracellular lactate concentration induced by glutamate were tested with a lactate detection kit. RESULTS: Compared with the control group, treatment with 1 mmol/L glutamate decreased the astrocytes' maximal respiration and spare respiratory capacity but increased their glycolytic capacity and glycolytic reserve. Further analysis found that 1-h treatment with different concentrations of glutamate (0.1-1 mmol/L) increased lactate release from astrocytes, however the cell viability was not affected by the glutamate treatment. CONCLUSION: The current study provided direct evidence that exogenous glutamate treatment impaired the mitochondrial respiration capacity of astrocytes and enhanced aerobic glycolysis, which could be involved in glutamate injury or protection mechanisms in response to neurological disorders.

[Determination of acidity and vitamin C in apples using portable NIR analyzer].

Near infrared (NIR) spectroscopy technology based on a portable NIR analyzer, combined with kernel Isomap algorithm and generalized regression neural network (GRNN) has been applied to establishing quantitative models for prediction of acidity and vitamin C in six kinds of apple samples. The obtained results demonstrated that the fitting and the predictive accuracy of the models with kernel Isomap algorithm were satisfactory. The correlation between actual and predicted values of calibration samples (R(c)) obtained by the acidity model was 0.999 4, and for prediction samples (R(p)) was 0.979 9. The root mean square error of prediction set (RMSEP) was 0.055 8. For the vitamin C model, R(c) was 0.989 1, R(p) was 0.927 2, and RMSEP was 4.043 1. Results proved that the portable NIR analyzer can be a feasible tool for the determination of acidity and vitamin C in apples.