Volatilisation Behaviour and Mechanism of Lead-Containing Slag during Physical Property Tests.

According to physical property tests of lead-containing slag, the volatilisation behaviour of lead slag will have adverse impacts on test accuracy and further affect the control of chemical reactions, solidification and removal of inclusions during smelting. To analyse the volatilisation characteristics of lead slag, in this paper, four kinds of lead slags from lead direct reduction smelting with different PbO and ZnO content are taken. thermogravimetry, ISP-TOF were used. Additionally, the changes in volatiles and slag composition and phases were analysed with XRD and ICPS, and the volatilisation reaction mechanism was discussed. The results indicated that the volatilisation of lead slag can lead to a big weight loss of about to the slag with higher PbO content. The weight loss increases with the PbO content in slag increases. The volatile corresponding to the weight loss above 900 °C is mainly PbO and less ZnO. The higher the temperature is, the stronger the volatilisation is. With the increase in temperature and keeping time, most of the PbO can be evaporated and leaves little PbO in the residual slag. This will has great effect to physico-chemical property measurement of the slag with higher PbO content, especially to the property measurement that be kept at high temperature for a long time. Because the volatiles is trend to condense with the temperature decrease, mass spectrometer is limited by the condensation of volatiles, i.e., PbO, ZnO and so on, in the connection pipeline. The device should be modified for this use.

Diagnostic accuracy of magnetically guided capsule endoscopy with a detachable string for detecting oesophagogastric varices in adults with cirrhosis: prospective multicentre study.

OBJECTIVE: To evaluate the diagnostic accuracy and safety of using magnetically guided capsule endoscopy with a detachable string (ds-MCE) for detecting and grading oesophagogastric varices in adults with cirrhosis. DESIGN: Prospective multicentre diagnostic accuracy study. SETTING: 14 medical centres in China. PARTICIPANTS: 607 adults (>18 years) with cirrhosis recruited between 7 January 2021 and 25 August 2022. Participants underwent ds-MCE (index test), followed by oesophagogastroduodenoscopy (OGD, reference test) within 48 hours. The participants were divided into development and validation cohorts in a ratio of 2:1. MAIN OUTCOME MEASURES: The primary outcomes were the sensitivity and specificity of ds-MCE in detecting oesophagogastric varices compared with OGD. Secondary outcomes included the sensitivity and specificity of ds-MCE for detecting high risk oesophageal varices and the diagnostic accuracy of ds-MCE for detecting high risk oesophagogastric varices, oesophageal varices, and gastric varices. RESULTS: ds-MCE and OGD examinations were completed in 582 (95.9%) of the 607 participants. Using OGD as the reference standard, ds-MCE had a sensitivity of 97.5% (95% confidence interval 95.5% to 98.7%) and specificity of 97.8% (94.4% to 99.1%) for detecting oesophagogastric varices (both P<0.001 compared with a prespecified 85% threshold). When using the optimal 18% threshold for luminal circumference of the oesophagus derived from the development cohort (n=393), the sensitivity and specificity of ds-MCE for detecting high risk oesophageal varices in the validation cohort (n=189) were 95.8% (89.7% to 98.4%) and 94.7% (88.2% to 97.7%), respectively. The diagnostic accuracy of ds-MCE for detecting high risk oesophagogastric varices, oesophageal varices, and gastric varices was 96.3% (92.6% to 98.2%), 96.9% (95.2% to 98.0%), and 96.7% (95.0% to 97.9%), respectively. Two serious adverse events occurred with OGD but none with ds-MCE. CONCLUSION: The findings of this study suggest that ds-MCE is a highly accurate and safe diagnostic tool for detecting and grading oesophagogastric varices and is a promising alternative to OGD for screening and surveillance of oesophagogastric varices in patients with cirrhosis. TRIAL REGISTRATION: ClinicalTrials.gov NCT03748563.

SINE-Associated LncRNA SAWPA Regulates Porcine Zygotic Genome Activation.

In mice, retrotransposon-associated long noncoding RNAs (lncRNA) play important regulatory roles in pre-implantation development; however, it is largely unknown whether they function in the pre-implantation development in pigs. The current study aims to screen for retrotransposon-associated lncRNA in porcine early embryos and identifies a porcine 8-cell embryo-specific SINE-associated nuclear long noncoding RNA named SAWPA. SAWPA is essential for porcine embryonic development as depletion of SAWPA results in a developmental arrest at the 8-cell stage, accompanied by the inhibition of the JNK-MAPK signaling pathway. Mechanistically, SAWPA works in trans as a transcription factor for JNK through the formation of an RNA-protein complex with HNRNPA1 and MED8 binding the SINE elements upstream of JNK. Therefore, as the first functional SINE-associated long noncoding RNAs in pigs, SAWPA provides novel insights for the mechanism research on retrotransposons in mammalian pre-implantation development.

Effects of Tao Hong Si Wu decoction on IncRNA expression in rats with occlusion of middle cerebral artery / 中国药理学通报

Aim To screen and study the expression of long non-coding RNA (IncRNA) in rats with middle cerebral artery occlusion (MCAO) with MCAO treated with Tao Hong Si Wu decoction (THSWD) and determine the possible molecular mechanism of THSWD in treating MCAO rats. Methods Three cerebral hemisphere tissue were obtained from the control group, MCAO group and MCAO + THSWD group. RNA sequencing technology was used to identify IncRNA gene expression in the three groups. THSWD-regulated IncRNA genes were identified, and then a THSWD-regu-lated IncRNA-mRNA network was constructed. MCODE plug-in units were used to identify the modules of IncRNA-mRNA networks. Gene ontology (GO) and kyoto encyclopedia of genes and genomes (KEGG) were used to analyze the enriched biological functions and signaling pathways. Cis- and trans-regulatory genes for THSWD-regulated IncRNAs were identified. Reverse transcription real-time quantitative pol-ymerase chain reaction (RT-qPCR) was used to verify IncRNAs. Molecular docking was used to identify IncRNA-mRNA network targets and pathway-associated proteins. Results In MCAO rats, THSWD regulated a total of 302 IncRNAs. Bioinformatics analysis suggested that some core IncRNAs might play an important role in the treatment of MCAO rats with THSWD, and we further found that THSWD might also treat MCAO rats through multiple pathways such as IncRNA-mRNA network and network-enriched complement and coagulation cascades. The results of molecular docking showed that the active compounds gallic acid and a-mygdalin of THSWD had a certain binding ability to protein targets. Conclusions THSWD can protect the brain injury of MCAO rats through IncRNA, which may provide new insights for the treatment of ischemic stroke with THSWD.

Prediction of axillary lymph node metastasis in early breast cancer patients with ultrasonic videos based deep learning.

Objective: To develop a deep learning (DL) model for predicting axillary lymph node (ALN) metastasis using dynamic ultrasound (US) videos in breast cancer patients. Methods: A total of 271 US videos from 271 early breast cancer patients collected from Xiang'an Hospital of Xiamen University andShantou Central Hospitabetween September 2019 and June 2021 were used as the training, validation, and internal testing set (testing set A). Additionally, an independent dataset of 49 US videos from 49 patients with breast cancer, collected from Shanghai 10th Hospital of Tongji University from July 2021 to May 2022, was used as an external testing set (testing set B). All ALN metastases were confirmed using pathological examination. Three different convolutional neural networks (CNNs) with R2 + 1D, TIN, and ResNet-3D architectures were used to build the models. The performance of the US video DL models was compared with that of US static image DL models and axillary US examination performed by ultra-sonographers. The performances of the DL models and ultra-sonographers were evaluated based on accuracy, sensitivity, specificity, and area under the receiver operating characteristic curve (AUC). Additionally, gradient class activation mapping (Grad-CAM) technology was also used to enhance the interpretability of the models. Results: Among the three US video DL models, TIN showed the best performance, achieving an AUC of 0.914 (95% CI: 0.843-0.985) in predicting ALN metastasis in testing set A. The model achieved an accuracy of 85.25% (52/61), with a sensitivity of 76.19% (16/21) and a specificity of 90.00% (36/40). The AUC of the US video DL model was superior to that of the US static image DL model (0.856, 95% CI: 0.753-0.959, P<0.05). The Grad-CAM technology confirmed the heatmap of the model, which highlighted important subregions of the keyframe for ultra-sonographers' review. Conclusion: A feasible and improved DL model to predict ALN metastasis from breast cancer US video images was developed. The DL model in this study with reliable interpretability would provide an early diagnostic strategy for the appropriate management of axillary in the early breast cancer patients.

DynamicSleepNet: a multi-exit neural network with adaptive inference time for sleep stage classification.

Sleep is an essential human physiological behavior, and the quality of sleep directly affects a person's physical and mental state. In clinical medicine, sleep stage is an important basis for doctors to diagnose and treat sleep disorders. The traditional method of classifying sleep stages requires sleep experts to classify them manually, and the whole process is time-consuming and laborious. In recent years, with the help of deep learning, automatic sleep stage classification has made great progress, especially networks using multi-modal electrophysiological signals, which have greatly improved in terms of accuracy. However, we found that the existing multimodal networks have a large number of redundant calculations in the process of using multiple electrophysiological signals, and the networks become heavier due to the use of multiple signals, and difficult to be used in small devices. To solve these two problems, this paper proposes DynamicSleepNet, a network that can maximize the use of multiple electrophysiological signals and can dynamically adjust between accuracy and efficiency. DynamicSleepNet consists of three effective feature extraction modules (EFEMs) and three classifier modules, each EFEM is connected to a classifier. Each EFEM is able to extract signal features while making the effective features more prominent and the invalid features are suppressed. The samples processed by the EFEM are given to the corresponding classifier for classification, and if the classifier considers the uncertainty of the sample to be below the threshold we set, the sample can be output early without going through the whole network. We validated our model on four datasets. The results show that the highest accuracy of our model outperforms all baselines. With accuracy close to baselines, our model is faster than the baselines by a factor of several to several tens, and the number of parameters of the model is lower or close. The implementation code is available at: https://github.com/Quinella7291/A-Multi-exit-Neural-Network-with-Adaptive-Inference-Time-for-Sleep-Stage-Classification/.

Clinical Comparison of the Efficacy of Systemic Thrombolysis,Catheter-Directed Thrombolysis,and AngioJet Percutaneous Mechanical Thrombectomy in Acute Lower Extremity Deep Venous Thrombosis / 中国医学科学院学报

Objective To compare the clinical effects of three treatment methods including systemic thrombolysis(ST),catheter-directed thrombolysis(CDT),and AngioJet percutaneous mechanical thrombectomy(PMT)in acute lower extremity deep venous thrombosis(LEDVT). Methods The data of 82 patients diagnosed with LEDVT in the Department of Vascular and Gland Surgery of the First Affiliated Hospital of Hebei North University from January 2017 to December 2020 were collected.The patients were assigned into a ST group(n=50),a CDT group(n=16),and a PMT group(n=16)according to different treatment methods.The efficacy and safety were compared among the three groups. Results Compared with that before treatment,the circumferential diameter difference of both lower limbs on days 1,2,and 3 of treatment in the ST,CDT,and PMT groups reduced(all P<0.001).The PMT group showed smaller circumferential diameter difference of lower limbs on days 1,2,and 3 of treatment than the ST group(all P<0.001)and smaller circumferential diameter difference of the lower patellar margin on day 1 of treatment than the CDT group(P<0.001).The PMT group showed higher diminution rate for swelling of the affected limb at the upper and lower edges of the patella than the ST group(P<0.001)and higher diminution rate for swelling at the upper edge of the patella than the CDT group(P=0.026).The incidence of complications after treatment showed no significant differences among the three groups(all P>0.05).The median of hospital stay in the PMT group was shorter than that in the ST and CDT groups(P=0.002,P=0.001).The PMT group had higher thrombus clearance rate than the ST group(P=0.002)and no significant difference in the thrombus clearance rate from the CDT group(P=0.361).The vascular recanalization rates in the PMT(all P<0.001)and CDT(P<0.001,P=0.002,P=0.009)groups 3,6,and 12 months after treatment were higher than those in ST group,and there were no significant differences between PMT and CDT groups(P=0.341,P=0.210,P=0.341). Conclusions ST,CDT,and PMT demonstrated significant efficacy in the treatment of LEDVT,and PMT was superior to ST and CDT in terms of circumferential diameter difference of the lower limbs,diminution rate for swelling of the affected limb,thrombus clearance rate,length of hospital stay,and long-term vascular recanalization.There was no obvious difference in safety among the three therapies.

Tannin Reduces the Incidence of Polyspermic Penetration in Porcine Oocytes.

Tannin (TA) improves porcine oocyte cytoplasmic maturation and subsequent embryonic development after in vitro fertilization (IVF). However, the mechanism through which TA blocks polyspermy after IVF remains unclear. Hence, the biological function of organelles (cortical granule [CG], Golgi apparatus, endoplasmic reticulum [ER], and mitochondria) and the incidence of polyspermic penetration were examined. We found no significant difference in oocyte nuclear maturation among the 1 µg/mL, 10 µg/mL TA, and control groups. Moreover, 100 µg/mL TA significantly reduced 1st polar body formation rate compared to the other groups. Additionally, 1 and 10 µg/mL TA significantly increased the protein levels of GDF9, BMP15, and CDK1 compared to the control and 100 µg/mL TA groups. Interestingly, 1 and 10 µg/mL TA improved the normal distribution of CGs, Golgi, ER, and mitochondria by upregulating organelle-related gene expression and downregulating ER stress (CHOP) gene expression. Simultaneously, 1 and 10 µg/mL TA significantly increased the proportion of normal fertilized oocytes (2 pronuclei; 2 PN) and blastocyst formation rate compared to the control, as well as that of 100 µg/mL TA after IVF by upregulating polyspermy-related genes. In conclusion, TA during IVM enhances 2PN and blastocyst formation rates by regulating organelles' functions and activities.

GW842166X Alleviates Osteoarthritis by Repressing LPS-mediated Chondrocyte Catabolism in Mice.

OBJECTIVE: To explore the role and underlying mechanism of GW842166X on osteoarthritis and osteoarthritis-associated abnormal catabolism. METHODS: The extracted mouse chondrocytes were treated with GW842166X followed by lipopolysaccharide (LPS). The chondrocytes were divided into the control group, LPS group, LPS+50 nmol/L GW842166X group, and LPS+100 nmol/L GW842166X group. The cytotoxicity of GW842166X was tested using the CCK-8 assay. Western blot, RT-qPCR, and ELISA were applied to evaluate the expression of the inflammatory biomarkers in mouse chondrocytes. The expression of extracellular matrix molecules was detected by the Western blot, RT-qPCR, and immunofluorescence. Additionally, the activity of NF-κB was checked by the Western blot and immunofluorescence. The mouse Hulth models were generated to examine the in vivo effects of GW842166X on osteoarthritis. Hematoxylin and eosin staining, safranin O/fast green staining, and immunohistochemistry were applied to detect the histological changes. RESULTS: GW842166X below 200 µmol/L had no cytotoxicity on the mouse chondrocytes. LPS-induced high expression of TGF-ß1, IL-10, TNF-α, and IL-6 was significantly reduced by GW842166X. In addition, GW842166X upregulated the expression of aggrecan and collagen type III, which was downregulated after the LPS stimulation. The upregulated expression of ADAMTS-5 and MMP-13 by LPS stimulation was dropped in response to the GW842166X treatment. Furthermore, LPS decreased the IκBα expression in the cytoplasm and increased the nuclear p65 expression. However, these changes were reversed by the GW842166X pretreatment. Moreover, the damages in the knees caused by the Hulth surgery in mice were restored by GW842166X. CONCLUSION: GW842166X impeded the LPS-mediated catabolism in mouse chondrocytes, thereby inhibiting the progression of osteoarthritis.

Ramelteon Reduces Oxidative Stress by Maintenance of Lipid Homeostasis in Porcine Oocytes.

This study aimed to determine the underlying mechanism of ramelteon on the competence of oocyte and subsequent embryo development in pigs during in vitro maturation (IVM). Our results showed that the cumulus expansion index was significantly lower in the control group compared to the ramelteon groups (p < 0.05). Moreover, supplementation of 10−11 and 10−9 M ramelteon significantly increased the cumulus expansion and development-related genes expression, and reduced apoptosis in cumulus cells (p < 0.05). In oocytes, the nuclear maturation rate was significantly improved in 10−11, 10−9, and 10−7 M ramelteon groups compared to the control (p < 0.05). Additionally, the level of intracellular GSH was significantly increased and ROS was significantly decreased in ramelteon-supplemented groups, and the gene expression of oocyte development and apoptosis were significantly up- and down-regulated by 10−11 and 10−9 M ramelteon (p < 0.05), respectively. The immunofluorescence results showed that the protein levels of GDF9, BMP15, SOD1, CDK1, and PGC1α were significantly increased by 10−11 M ramelteon compared to the control (p < 0.05). Although there was no significant difference in cleavage rate, the blastocyst formation rate, total cell numbers, and hatching/-ed rate were significantly improved in 10−11 M ramelteon group compared to the control (p < 0.05). Furthermore, embryo development, hatching, and mitochondrial biogenesis-related genes were dramatically up-regulated by 10−11 M ramelteon (p < 0.05). In addition, the activities of lipogenesis and lipolysis in oocytes were dramatically increased by 10−11 M ramelteon compared to the control (p < 0.05). In conclusion, supplementation of 10−11 M ramelteon during IVM improved the oocyte maturation and subsequent embryo development by reducing oxidative stress and maintenance of lipid homeostasis.

The comparison between the effect of Glycyrrhizae uralensis (Gan-Cao) and Montelukast on the expression of T-bet and GATA-3 genes in children with allergic asthma.

Pediatric allergic asthma is a chronic disease that affects the lungs and airways. If a child is exposed to certain stimulants such as pollen inhalation, colds, or respiratory infections, the lungs become inflamed and if left untreated can lead to dangerous asthma attacks. One of the most important treatments for this disease is the use of leukotriene modulators, such as montelukast. But recently, due to easier access, cheaper prices and fewer side effects, attention has shifted to non-chemical treatments. Gan-Cao (Glycyrrhizae uralensis), as traditional Chinese medicine, has been proved to have a good therapeutic effect on experimental allergic asthma. But its anti-asthma mechanism is currently unclear. Therefore, the study aimed the comparison between the effect of Gan-Cao and montelukast on the expression of T-bet and GATA-3 genes in children with allergic asthma. For this purpose, fifty children with allergic asthma were divided into two groups. The first group was treated with montelukast for one month. The second group was treated with Gan-Cao root extract. Then the peripheral blood mononuclear cells were isolated, their RNA was extracted, and the relative expression of T-bet and GATA3 transcription factors was evaluated by Real-time PCR. The relationship between them and risk factors for asthma was assessed by relevant statistical tests. The result showed the expression of the GATA3 gene (P = 0.102), T-bet gene (P = 0.888), and the expression ratio of T-bet/GATA-3 genes (P = 0.061) was not significantly different between the two groups. It showed that Gan-Cao can affect the expression of these genes just as much as montelukast. Therefore, this Chinese herb can be used as an alternative or supplement medicine to treat allergic asthma in children.

Melatonin Regulates Lipid Metabolism in Porcine Cumulus-Oocyte Complexes via the Melatonin Receptor 2.

Previous studies suggest that the inclusion of melatonin (MTn) in in vitro maturation protocols improves the developmental competence of oocytes by scavenging reactive oxygen species (ROS). However, the molecular mechanisms integrating melatonin receptor (MT)-mediated lipid metabolism and redox signaling during in vitro cumulus-oocyte complex (COC) development still remain unclear. Here, we aimed to elucidate the potential role of MTn receptors in lipid metabolic adjustments during in vitro porcine COC development. We observed that MTn-mediated Gsα-cAMP/PKA signaling facilitated lipolysis primarily through the MT2 receptor and subsequently increased fatty acid (FA) release by hydrolyzing intracellular triglycerides (TGs) in cumulus cells. Furthermore, CD36 was a critical FA transporter that transported available FAs from cumulus cells to oocytes and promoted de novo TG synthesis in the latter. In addition, MTn regulated lipogenesis and intracellular lipolysis to maintain lipid homeostasis and limit ROS production, thereby supporting oocyte cytoplasmic maturation and the subsequent embryo development. Taken together, these findings provide insight into the possible mechanism integrating MT2-mediated lipid homeostasis and redox signaling, which limits ROS production during in vitro COC development. Therefore, understanding the dynamics of the interactions between lipid homeostasis and redox signaling driven by MT2 is necessary in order to predict drug targets and the effects of therapeutics used to improve female reproductive health.

Ubiquitin⁃like Protein UBQLN2 Exerts Tumor Suppressor Effect by Inhibiting Activation of Wnt Signaling Pathway in Colon Cancer / 中国生物化学与分子生物学报

Colon cancer is a malignant tumor that harms human health. In spite of significant progress in our understanding of the molecular pathogenesis, the prognosis for patients with colon cancer remains poor. Ubiquilin-2 (UBQLN2) is a member of the ubiquitin family, actively implicated in the degradation of misfolded and redundant proteins through the ubiquitin-proteasome system and macroautophagy. However, the role and mechanism of UBQLN2 in colon cancer are not clear. This study was designed to explore the role of UBQLN2 in colon cancer and whether the Wnt pathway is involved. IHC and Western blotting analysis showed lower UBQLN2 expression in colon cancer tissues and cells (P<0. 05), and exhibited a negative correlation of UBQLN2 expression with clinical stage and lymph node metastasis (P< 0. 05). CCK-8 assay and flow cytometry demonstrated that UBQLN2 expression could inhibit colon cancer cell proliferation and promote cell apoptosis (P<0. 05). Western blotting analysis reveals that suppression of UBQLN2 enhances Bcl-2 expression but inhibits Bax and Wnt signaling. Taken together, these results showed that UBQLN2 inhibits proliferation and promotes apoptosis of colorectal cancer cells through suppressing the Wnt signaling pathway.

The epidemiological characteristics of pneumoconiosis in Qingyuan City from 1949 to 2018 / 中华劳动卫生职业病杂志

Objective: In order to find out the prevalence and death of pneumoconiosis in Qingyuan City, to explore the regularity of pneumoconiosis and lay a foundation for the prevention and management of pneumoconiosis. Methods: In August to December 2019, the basic data of pneumoconiosis from 1949 to 2018 were obtained through the monitoring of death causes of residents, occupational disease management system, Guangdong population information system and other means. The reported cases of pneumoconiosis were followed up, and retrospective investigation was conducted to analyze the basic conditions, the length of service exposed to dust, the time of diagnosis, the type of disease, the stage and the combined status of tuberculosis of pneumoconiosis cases. Results: From 1949 to 2018, a total of 466 cases of new pneumoconiosis were reported in Qingyuan City, including 325 cases of death (69.74%) , 114 cases of survival (24.46%) and 27 cases of loss of follow-up (5.80%) . The cases were mainly concentrated in the age group of 40-89 years (80.04%, 373/466) . There were 411 male cases (88.20%) and 7 female cases (1.50%) . The median length of service exposed to dust was 10.7 (6.0, 16.0) years. The diagnosis time of pneumoconiosis cases was mainly from 1949 to 1986 (68.67%, 320/466) , and the death cases were mainly from 1949 to 1986 (82.77%, 269/325) . Silicosis was the main type of pneumoconiosis (398 cases, 85.41%) . 200 cases (42.92%) were diagnosed as stage I pneumoconiosis, 185 cases (39.69%) were stage II pneumoconiosis, 81 cases (17.38%) were stage III pneumoconiosis at the first diagnosis. 102 cases (21.89%) were pneumoconiosis with tuberculosis. The proportion of death and lost follow-up cases diagnosed as stage I pneumoconiosis for the first time was significantly lower than that of survival cases, and the proportion diagnosed as stage II pneumoconiosis and the complication rate of tuberculosis were significantly higher than those of survival cases (χ(2)=15.48, 11.29, 32.73, P<0.001) . Conclusion: Pneumoconiosis in Qingyuan City is mainly silicosis. The number of new cases has been increasing in the past decade, and the prevention and control situation is still severe. The comprehensive prevention and treatment of silica dust should be included in the focus of supervision of government functional departments.

A real-time and modular approach for quick detection and mechanism exploration of DPIs with different carrier particle sizes

Dry powder inhalers (DPIs) had been widely used in lung diseases on account of direct pulmonary delivery, good drug stability and satisfactory patient compliance. However, an indistinct understanding of pulmonary delivery processes (PDPs) hindered the development of DPIs. Most current evaluation methods explored the PDPs with over-simplified models, leading to uncompleted investigations of the whole or partial PDPs. In the present research, an innovative modular process analysis platform (MPAP) was applied to investigate the detailed mechanisms of each PDP of DPIs with different carrier particle sizes (CPS). The MPAP was composed of a laser particle size analyzer, an inhaler device, an artificial throat and a pre-separator, to investigate the fluidization and dispersion, transportation, detachment and deposition process of DPIs. The release profiles of drug, drug aggregation and carrier were monitored in real-time. The influence of CPS on PDPs and corresponding mechanisms were explored. The powder properties of the carriers were investigated by the optical profiler and Freeman Technology four powder rheometer. The next generation impactor was employed to explore the aerosolization performance of DPIs. The novel MPAP was successfully applied in exploring the comprehensive mechanism of PDPs, which had enormous potential to be used to investigate and develop DPIs.

Tannin Supplementation Improves Oocyte Cytoplasmic Maturation and Subsequent Embryo Development in Pigs.

To investigate the effects of tannins (TA) on porcine oocyte in vitro maturation (IVM), different concentrations of TA (0, 1, 10 and 100 µg/mL) were supplemented with a maturation medium and the COCs and subsequent embryonic development were examined. The results showed that 10 µg/mL TA significantly improved the cumulus expansion index (CEI), cumulus-expansion-related genes (PTGS1, PTGS2, PTX-3, TNFAIP6 and HAS2) expression and blastocyst formation rates after parthenogenetic activation (PA), in vitro fertilization (IVF) and somatic cell nuclear transfer (SCNT) compared to the control groups, but not oocyte nuclear maturation. Nevertheless, 10 µg/mL TA dramatically enhanced the mRNA expression of oocyte-development-related genes (BMP15, GDF9, CDC2 and CYCLIN B1), GSH, ATP, SOD1, PGC1α, BMP15, GDF9 and CDC2 levels and reduced intracellular ROS level in porcine oocytes. These results indicated that porcine oocyte cytoplasmic maturation was improved by 10 µg/mL TA treatment during IVM. In contrast, a high concentration of TA (100 µg/mL) significantly decreased the CEI and PTGS1, PTGS2, PTX-3 and HAS2 mRNA expressions in cumulus cells, and reduced oocyte nuclear maturation and the total cell numbers/blastocyst. In general, these data showed that 10 µg/mL TA supplementation has beneficial effects on oocyte cytoplasmic maturation and subsequent embryonic development in pigs.

Metabolomics provide new insights into mechanisms of Wolbachia-induced paternal defects in Drosophila melanogaster.

Wolbachia is a group of intracellular symbiotic bacteria that widely infect arthropods and nematodes. Wolbachia infection can regulate host reproduction with the most common phenotype in insects being cytoplasmic incompatibility (CI), which results in embryonic lethality when uninfected eggs fertilized with sperms from infected males. This suggests that CI-induced defects are mainly in paternal side. However, whether Wolbachia-induced metabolic changes play a role in the mechanism of paternal-linked defects in embryonic development is not known. In the current study, we first use untargeted metabolomics method with LC-MS to explore how Wolbachia infection influences the metabolite profiling of the insect hosts. The untargeted metabolomics revealed 414 potential differential metabolites between Wolbachia-infected and uninfected 1-day-old (1d) male flies. Most of the differential metabolites were significantly up-regulated due to Wolbachia infection. Thirty-four metabolic pathways such as carbohydrate, lipid and amino acid, and vitamin and cofactor metabolism were affected by Wolbachia infection. Then, we applied targeted metabolomics analysis with GC-MS and showed that Wolbachia infection resulted in an increased energy expenditure of the host by regulating glycometabolism and fatty acid catabolism, which was compensated by increased food uptake. Furthermore, overexpressing two acyl-CoA catabolism related genes, Dbi (coding for diazepam-binding inhibitor) or Mcad (coding for medium-chain acyl-CoA dehydrogenase), ubiquitously or specially in testes caused significantly decreased paternal-effect egg hatch rate. Oxidative stress and abnormal mitochondria induced by Wolbachia infection disrupted the formation of sperm nebenkern. These findings provide new insights into mechanisms of Wolbachia-induced paternal defects from metabolic phenotypes.

Failure to maintain full-term pregnancies in pig carrying klotho monoallelic knockout fetuses.

BACKGROUND: Small animals that show a deficiency in klotho exhibit extremely shortened life span with multiple aging-like phenotypes. However, limited information is available on the function of klotho in large animals such as pigs. RESULTS: In an attempt to produce klotho knockout pigs, an sgRNA specific for klotho (targeting exon 3) was designed and Cas9-sgRNA ribonucleoproteins were transfected into porcine fibroblasts. Transfected fibroblasts were cultured for one to 2 days and then directly used for nuclear transfer without selection. The cloned embryos were cultured in vitro for 7 days and analyzed to detect modifications of the klotho gene by both T7E1 and deep sequencing analysis. Modification succeeded in 13 of 20 blastocysts (65%), 8 of which (40.0%) were monoallelic modifications and 5 (25.0%) were biallelic modifications. Based on high mutation rates in blastocysts, we transferred the cloned embryos to 5 recipient pigs; 1 recipient was pregnant and 16 fetuses were recovered at Day 28 post transfer. Of the 16 fetuses, 9 were resorbing and 7 were viable. Four of 9 (44.4%) resorbing fetuses and 3 of the 7 (42.9%) viable fetuses had monoallelic modifications. Thus, 3 klotho monoallelic knockout cell lines were established by primary culture. A total of 2088 cloned embryos reconstructed with 2 frame-shifted cell lines were transferred to 11 synchronized recipients. Of the recipients, 7 of 11 eleven (63.6%) became pregnant. However, none of the pregnancies was maintained to term. To discover why klotho monoallelic knockout fetuses were aborted, expression of aging- and apoptosis-related genes and klotho protein in placentas from klotho monoallelic knockout and wild-type fetuses was investigated. Placentas from klotho monoallelic knockout fetuses showed negatively changed expression of aging- and apoptosis-related genes with lower relative expression of klotho protein. These results indicated that the reason why klotho monoallelic knockout fetuses were not maintained to term was possibly due to decreased klotho expression in placentas, negatively affecting aging- and apoptosis-related genes. CONCLUSIONS: Klotho monoallelic knockout porcine fetal fibroblasts were successfully established. However, pigs carrying klotho monoallelic knockout fetuses failed to maintain full-term pregnancy and a decrease in klotho expression in placenta likely leads to pregnancy loss.

Effect of Bushen Huatan Prescription on Serum LPS and TLR4/MyD88/NF-κB Signaling Pathway in Rats with Ovariectomy-induced Osteoporosis / 中国实验方剂学杂志

Objective:To explore the effect of Bushen Huatan prescription on serum lipopolysaccharide （LPS） and Toll-like receptor 4 （TLR4）/ myeloid cell differentiation protein 88 （MyD88）/nuclear transcription factor-<italic>κ</italic>B （NF-<italic>κ</italic>B） signaling pathway in rats with ovariectomy-induced osteoporosis. Method:Sixty SPF 6-month-old female rats were randomly divided into sham operation group， model group， estradiol valerate group and Bushen Huatan prescription low， medium and high dose groups.One week after modeling by bilateral ovariectomy， 8 rats in each group were selected to receive intragastric administration.The estradiol valerate group was given 0.184 mg·kg^-1 by gavage， and Bushen Huatan prescription low， middle and high dose groups were given 4.7， 9.4 and 18.8 g·kg^-1 by gavage， sham operation group and model group were given 0.9% saline 4 mL by gavage respectively.After 12 weeks of intervention， the rats were sacrificed for detection.Serum LPS was detected by enzyme linked immunosorbent assay （ELISA）， while protein expressions of TLR4， MyD88 and phosphorylated （p）-NF-<italic>κ</italic>B p65 in bone tissue were detected by Western blot， and the mRNA expressions of TLR4， MyD88， NF-<italic>κ</italic>B p65， IL-1<italic>β</italic>， and IL-6 in bone tissue were detected by quantitative real-time polymerase chain reaction（PCR）. Result:Compared with sham operation group， the serum LPS level as well as protein expression of TLR4， MyD88， p-NF-<italic>κ</italic>B p65 and mRNA expression of TLR4， MyD88， NF-<italic>κ</italic>B p65， IL-1<italic>β</italic>， and IL-6 significantly increased in model group（<italic>P</italic><0.05）.Compared with the model group， serum LPS level， protein expression of TLR4， MyD88， and p-NF-<italic>κ</italic>B p65， mRNA levels of TLR4， MyD88， and NF-<italic>κ</italic>B p65 in bone tissues as well as downstream inflammatory factors IL-1<italic>β</italic>， IL-6 mRNA expression decreased to different degrees in estradiol valerate group and Bushen Huatan prescription high dose group（<italic>P</italic><0.05）. Conclusion:Bushen Huatan prescription can reduce serum LPS content， regulate mRNA and protein expression of TLR4， MyD88， NF-<italic>κ</italic>B p65 and p-NF-<italic>κ</italic>B p65 in TLR4/MyD88/NF-<italic>κ</italic>B pathway， and down-regulate mRNA levels of IL-1<italic>β</italic> and IL-6 in bone tissues to improve bone microstructure and inhibit the development of postmenopausal osteoporosis （PMOP）.