Identification and bioactivity evaluation of ring-opening and lactone forms of enterolactone from sheep fed flaxseed cake.

The previously undescribed lactone ring-opening enterolactone and its sulphate were purified along with the lactone counterparts from the urine of dairy sheep fed flaxseed cake. The structures were determined by NMR and MS analyses. The ring-opening and lactone forms underwent mutual transformation with changes in pH and milk could protect the lactone form. Enterolactone exhibited more effective anti-proliferation activity on MDA-MB-231 breast cancer cells than its ring-opening counterpart, while the ring-opening enterolactone demonstrated more effective anti-osteoporosis activity than the lactone form. The results indicated the potential for targeting biological functions through pH and medium manipulation.

Anti-osteoporosis effects of mammalian lignans and their precursors from flaxseed and safflower seed using zebrafish model.

Secoisolariciresinol diglucoside (SDG) and tracheloside (TCL) are the main lignan components of flaxseed cake and safflower seed cake, which are by-products of oil extraction. Both SDG and TCL are metabolized into mammalian lignan enterolactone (EL) with the involvement of intestinal bacteria. In this research, we evaluated the anti-osteoporosis effects of SDG and the in vivo metabolites EL and enterodiol (ED) prepared in our previous work, as well as the newly isolated chemical constituents from safflower seed, including TCL, the lactone ring opening product of TCL (OTCL) and two alkaloids on the alloxan-induced zebrafish model. All the compounds showed significant anti-osteoporosis effects at 80 µM, with p < 0.05 for EL and p < 0.001 for other compounds compared with the model. SDG and TCL showed the most significant and concentration-dependent effects, with p < 0.001 compared with model at 20 µM. The alkaloids, N-coumaroylserotonin glucoside and N-feruloylserotonin glucoside, also showed anti-osteoporosis at 20 µM with p < 0.01, whereas EL, ED, and OTCL showed no significant effects. Quantitative real-time polymerase chain reaction revealed that SDG and TCL upregulated the expression of osteogenic genes Runx2, SP7, OPG, Col1a1a, Alp, ON, OPN, and OCN in alloxan-treated zebrafish. The in vivo metabolite of lignans, EL, showed significant anti-inflammatory effect (p < 0.01) at 20 µM, which might also help to combat osteoporosis and other complications caused by excessive immune response in the body. The results provided scientific data for using the oil extraction by-products as sources of anti-osteoporosis compounds. PRACTICAL APPLICATION: This study found that lignans in flaxseed cake and safflower seed cake exhibited anti-osteoporosis effects by upregulating the expression of osteogenic genes, making the oil extraction by-products sources of anti-osteoporosis compounds.

Effects and mechanisms of 6-hydroxykaempferol 3,6-di-O-glucoside-7-O-glucuronide from Safflower on endothelial injury in vitro and on thrombosis in vivo.

Background: The florets of Carthamus tinctorius L. (Safflower) is an important traditional medicine for promoting blood circulation and removing blood stasis. However, its bioactive compounds and mechanism of action need further clarification. Objective: This study aims to investigate the effect and possible mechanism of 6-hydroxykaempferol 3,6-di-O-glucoside-7-O-glucuronide (HGG) from Safflower on endothelial injury in vitro, and to verify its anti-thrombotic activity in vivo. Methods: The endothelial injury on human umbilical vein endothelial cells (HUVECs) was induced by oxygen-glucose deprivation followed by reoxygenation (OGD/R). The effect of HGG on the proliferation of HUVECs under OGD/R was evaluated by MTT, LDH release, Hoechst-33342 staining, and Annexin V-FITC apoptosis assay. RNA-seq, RT-qPCR, Enzyme-linked immunosorbent assay and Western blot experiments were performed to uncover the molecular mechanism. The anti-thrombotic effect of HGG in vivo was evaluated using phenylhydrazine (PHZ)-induced zebrafish thrombosis model. Results: HGG significantly protected OGD/R induced endothelial injury, and decreased HUVECs apoptosis by regulating expressions of hypoxia inducible factor-1 alpha (HIF-1α) and nuclear factor kappa B (NF-κB) at both transcriptome and protein levels. Moreover, HGG reversed the mRNA expression of pro-inflammatory cytokines including IL-1ß, IL-6, and TNF-α, and reduced the release of IL-6 after OGD/R. In addition, HGG exhibited protective effects against PHZ-induced zebrafish thrombosis and improved blood circulation. Conclusion: HGG regulates the expression of HIF-1α and NF-κB, protects OGD/R induced endothelial dysfunction in vitro and has anti-thrombotic activity in PHZ-induced thrombosis in vivo.

Regulation of wakefulness by GABAergic dorsal raphe nucleus-ventral tegmental area pathway.

The dorsal raphe nucleus (DRN) has previously been proved to be involved in the regulation of the sleep-wake behavior. DRN contains several neuron types, such as 5-HTergic and GABAergic neurons. GABAergic neurons, which are the second largest cell subtype in the DRN, participate in a variety of neurophysiological functions. However, their role in sleep-wake regulation and the underlying neural circuitry remains unclear. Herein, we used fiber photometry and synchronous electroencephalogram (EEG)/electromyography (EMG) recording to demonstrate that DRN GABAergic neurons exhibit high activities during wakefulness and low activities during NREM sleep. Short-term optogenetic activation of DRN GABAergic neurons reduced the latency of NREM-to-wake transition and increased the probability of wakefulness, while long-term optogenetic activation of these neurons significantly increased the amount of wakefulness. Chemogenetic activation of DRN GABAergic neurons increased wakefulness for almost 2 h and maintained long-lasting arousal. In addition, inhibition of DRN GABAergic neurons with chemogenetics caused a reduction in the amount of wakefulness. Finally, similar to the effects of activating the soma of DRN GABAergic neurons, optogenetic stimulation of their terminals in the ventral tegmental area (VTA) induced instant arousal and promoted wakefulness. Taken together, our results illustrated that DRN GABAergic neurons are vital to the induction and maintenance of wakefulness, which promote wakefulness through the GABAergic DRN-VTA pathway.

Serum Cystatin C as a predictor of acute kidney injury in neonates: a meta-analysis

Abstract Objective: The objective of this meta-analysis is to evaluate the diagnostic value of serum Cystatin C in acute kidney injury (AKI) in neonates Sources: PubMed, Embase, Cochrane Library, Web of Science, China National Knowledge Infrastructure (CNKI), and WanFang Database were searched to retrieve the literature related to the diagnostic value of Cystatin C for neonatal AKI from inception to May 10, 2021. Subsequently, the quality of included studies was determined using the QUADAS-2 tool. Stata 15.0 statistical software was used to calculate the combined sensitivity (SEN), specificity (SPE), positive likelihood ratio (PLR), negative likelihood ratio (NLR), and diagnostic odds ratio (DOR). Additionally, meta-regression analysis and subgroup analysis contributed to explore the sources of heterogeneity Summary of the findings: Twelve articles were included. The pooled sensitivity was 0.84 (95%CI: 0.74-0.91), the pooled specificity was 0.81 (95%CI: 0.75-0.86), the pooled PLR was 4.39 (95%CI: 3.23-5.97), the pooled NLR was 0.19 (95%CI: 0.11-0.34), and the DOR was 22.58 (95%CI: 10.44-48.83). The area under the receiver operating characteristic curve (AUC) was 0.88 (95%CI: 0.85-0.90). No significant publication bias was identified (p > 0.05).

Control of Behavioral Arousal and Defense by a Glutamatergic Midbrain-Amygdala Pathway in Mice.

In response to external threatening signals, animals evolve a series of defensive behaviors that depend on heightened arousal. It is believed that arousal and defensive behaviors are coordinately regulated by specific neurocircuits in the central nervous system. The ventral tegmental area (VTA) is a key structure located in the ventral midbrain of mice. The activity of VTA glutamatergic neurons has recently been shown to be closely related to sleep-wake behavior. However, the specific role of VTA glutamatergic neurons in sleep-wake regulation, associated physiological functions, and underlying neural circuits remain unclear. In the current study, using an optogenetic approach and synchronous polysomnographic recording, we demonstrated that selective activation of VTA glutamatergic neurons induced immediate transition from sleep to wakefulness and obviously increased the amount of wakefulness in mice. Furthermore, optogenetic activation of VTA glutamatergic neurons induced multiple defensive behaviors, including burrowing, fleeing, avoidance and hiding. Finally, viral-mediated anterograde activation revealed that projections from the VTA to the central nucleus of the amygdala (CeA) mediated the wake- and defense-promoting effects of VTA glutamatergic neurons. Collectively, our results illustrate that the glutamatergic VTA is a key neural substrate regulating wakefulness and defensive behaviors that controls these behaviors through its projection into the CeA. We further discuss the possibility that the glutamatergic VTA-CeA pathway may be involved in psychiatric diseases featuring with excessive defense.

Serum Cystatin C as a predictor of acute kidney injury in neonates: a meta-analysis.

OBJECTIVES: The objective of this meta-analysis is to evaluate the diagnostic value of serum Cystatin C in acute kidney injury (AKI) in neonates. SOURCES: PubMed, Embase, Cochrane Library, Web of Science, China National Knowledge Infrastructure (CNKI), and WanFang Database were searched to retrieve the literature related to the diagnostic value of Cystatin C for neonatal AKI from inception to May 10, 2021. Subsequently, the quality of included studies was determined using the QUADAS-2 tool. Stata 15.0 statistical software was used to calculate the combined sensitivity (SEN), specificity (SPE), positive likelihood ratio (PLR), negative likelihood ratio (NLR), and diagnostic odds ratio (DOR). Additionally, meta-regression analysis and subgroup analysis contributed to explore the sources of heterogeneity. SUMMARY OF THE FINDINGS: Twelve articles were included. The pooled sensitivity was 0.84 (95%CI: 0.74-0.91), the pooled specificity was 0.81 (95%CI: 0.75-0.86), the pooled PLR was 4.39 (95%CI: 3.23-5.97), the pooled NLR was 0.19 (95%CI: 0.11-0.34), and the DOR was 22.58 (95%CI: 10.44-48.83). The area under the receiver operating characteristic curve (AUC) was 0.88 (95%CI: 0.85-0.90). No significant publication bias was identified (p > 0.05). CONCLUSIONS: Serum Cystatin C has a good performance in predicting neonatal AKI; therefore, it can be used as a candidate biomarker after the optimal level is determined by large prospective studies.

The effect of Omaha System-based continuous nursing care on the psychological status, self-esteem, and quality of life in epileptic children.

OBJECTIVES: This study explored and analyzed the effect of Omaha System-based continual nursing care on the psychological status, self-esteem and quality of life in epileptic children. METHODS: 127 epileptic children hospitalized from March 2018 to September 2019 were recruited as the study cohort and stochastically divided into an observation group (n=65) and a control group (n=62). The control group children were given regular out-of-hospital guidance after discharge, and the observation-group were treated with Omaha System-based continuous nursing intervention in addition to the routine out-of-hospital guidance the control group underwent. The two groups' psychological statuses, self-esteem, sense of defect, and quality of life were compared both pre- and post-intervention. RESULTS: The SAS and SDS scores in the two groups after the intervention were significantly lower than their pre-intervention scores (P<0.05), and the scores in the observation group after the intervention were evidently lower than the scores in the control group (P<0.05). The two groups' post-intervention SES and FIS scores were significantly higher than their pre-intervention scores (P<0.05), and the scores in the observation group were notably lower than the scores in the control group (P<0.05). The post-intervention quality of life scores in the two groups were dramatically higher than the pre-intervention scores (P<0.05), and the quality of life scores in the observation group were notably higher than the quality of life scores in the control group (P<0.05). CONCLUSION: The Omaha System-based continual nursing care can efficaciously elevate the psychological status of children with epilepsy, improve their self-esteem, reduce their sense of self-defect, and contribute to the improvement of their quality of life, so it is worthy of clinical promotion and application.

Technical note: Quantification of lignans in the urine, milk, and plasma of flaxseed cake-fed dairy sheep.

Mammalian lignans are phytoestrogens with important bioactivities, and their concentrations in livestock milk may influence the health of consumers. This research aimed to establish a method to quantify multiple mammalian lignans in the biofluids of dairy sheep using ultra-HPLC-triple quadropole mass spectrometry with multiple-reaction monitoring. Secoisolariciresinol, 2-[(4-hydroxy-3-methoxyphenyl)methyl]-3-[(3-hydroxyphenyl)methyl]-1,4-butanediol, enterodiol (ED), enterolactone (EL), ED-sulfate (ED-S), and EL-sulfate (EL-S) were purified from the urine of flaxseed cake-fed dairy sheep. The structures of these lignans were identified by a combination of mass and nuclear magnetic resonance spectra. These purified lignans were used as standards to optimize their quantification conditions in urine, milk, and plasma of dairy sheep. On this basis, the lignan metabolites in biofluids were quantified. To improve analysis sensitivity, plasma and milk were pretreated with acetonitrile containing 1% formic acid and passed through a HybridSPE-PL 55261-U column (Supelco, Bellefonte, PA). The limit of quantification of the lignans ranged from 1.43 to 18.3 ng/mL in plasma, and from 1.01 to 18.7 ng/mL in milk. The linearity of the calibration curves ranged from their limit of quantification to at least 217 ng/mL in plasma, and 217 ng/mL in milk. Regression coefficient of the calibration curves were above 0.99 for secoisolariciresinol, 2-[(4-hydroxy-3-methoxyphenyl)methyl]-3-[(3-hydroxyphenyl)methyl]-1,4-butanediol, ED, EL, ED-S, and EL-S, indicating satisfactory relationships between the peak areas and concentrations in the quantification range. The relative concentrations of ED-glucuronide and EL-glucuronide (EL-G) in different biofluids were compared based on their chromatogram peak areas. The sheep plasma contained all forms of mammalian lignans (i.e., ED, EL, ED-S, EL-S, ED-glucuronide, and EL-G.); the urine contained ED, EL, ED-S, and EL-S; and the milk contained ED, EL, ED-S, EL-S, and EL-G. Milk-to-plasma concentration ratios of the mammalian lignans indicated that the free forms were more permeable than the sulfated conjugates. Mammalian lignans found in sheep plasma and milk may provide health benefits to the sheep and sheep-product consumers. The analytical method established in this work could be used to quantify mammalian lignans in livestock products.

Multidisciplinary diagnosis and treatment of anastomotic leakage in esophageal cancer and its effect on prognosis / 医学研究生学报

Objective Early diagnosis and effective treatment are the key to reduce the mortality and improve the prognosis of anastomotic leakage which involved many disciplines after esophageal cancer surgery. The aim of the study is to investigate the value of multidisciplinary which combined diagnosis and treatments in anastomotic leakage after esophageal cancer operations. Methods A total of 82 patients with anastomotic leakage after esophageal cancer surgery in our hospital from June 2013 to June 2018 . 43 patients from June 2013 to December 2015 were included in the control group. 39 patients with anastomotic leakage from January 2016 to June 2018 were used as the observation group. Patients in the control group were given routine diagnosis and treatment, while the observation group was given a multidisciplinary (thoracic surgery, imaging, general surgery, nutrition,etc.) joint diagnosis and treatment model. Observe general data, prognosis, and fistula healing time. Results The proportion of occultation in the observation group was significantly higher than that in the control group(38.5% vs 18.6%), and the difference was statistically significant (P<0.05). The healing time of anastomotic leakage was lower in the observation group[(17.24±3.62)d] than in the control group[(33.85±4.85)d](P<0.05). Conclusion Multidisciplinary combined diagnosis and treatment can improve the incidence of occult sputum and reduce the healing time of anastomotic leakage. It is worthy of clinical promotion.

The learning curve of Da Vinci robotic surgical system for radical surgery in esophageal cancer / 医学研究生学报

Objective At present, there are few studies about the learning curve of da Vinci robot system for surgical treatment of esophageal cancer. This study aimed to evaluate the surgical outcomes of Da Vinci robotic surgical system for radical surgery in esophageal cancer and explore the learning curve characteristics of thoracic surgeon.Methods Clinical data of 60 consecutive cases who received Da Vinci robot-assisted radical esophagectomy between January 2011 and December 2015 in Nanjing General Hospital were reviewed retrospectively. The cases were divided into 3 groups according to the operation period: group A (operation time: January 2011 to January 2013), group B (operation time: February 2013 to February 2014) and group C (operation time: March 2014 to December 2015). There were 20 cases in each group. The docking time, thoracic cavity anatomical separation time, total operation time and complication were compared. The learning curve of robotic surgery system were analyzedResults The operations of 60 patients were successfully completed without any transfer to thoracotomy or laparotomy. There was no statistically significant difference in the incidence of complications between the 3 groups (P>0.05). With the increase in the number of surgical cases, the total operation time of group A (\[450.7±99.1\]min), group B (\[299.7±57.1\]min) and group C (\[248.5±71.5\]min) was gradually reduced. The difference was statistically significant (P0.05) . There were significant differences in total operation time among the three groups(P<0.05).After 20 cases, the fitting curve appeared turning angle, and then the curve began to slow down. Combined with the total operation time, pleural and abdominal cavity anatomical separation time and robotic docking time, it can be seen that the learning curve of Da Vinci robot-assisted radical robotic surgery in esophageal cancer was 20 cases.Conclusion After 20 cases of study, the thoracic surgeon can master the Da Vinci robotic surgical system for the radical surgery of esophageal cancer.

The effect of preoperative prehabilitation on nutritional status and body composition of patients with esophageal cancer after operation / 肠外与肠内营养

Objective: To observe the efficacy of preoperative prehabilitation on nutritional status and body composition of patients with esophageal cancer after operation. Methods: A total of 100 patients with esophageal cancer were randomly divided into the study group A and the control group B. The study group received prehabilitation from the day they adimitted to hospital and the control group received common preoperative preparation. The preoperative baseline data and postoperative indicators including levels of serum albumin, prealbumin, transferrin were recorded the day admitted to hospital and the 1, 3, 7, 30th day after surgery. Record the prognosis of the patients (albumin supplementation, incidence of postoperative complications, ICU monitoring time, total cost of hospitalization). And the body composition was examined on the day admitted to hospital and the 7, 30th day after surgery with BIA. Results: The level of serum albumin, prealbumin, transferrin and body composition were no statistically significant difference between the study group and the control group. Compared with the control group, the level of prealbumin were significantly higher in the study group on the 1, 3, 7, 30th day after surgery. The level of serum albumin, transferrin were also significantly higher in study group on the 3, 7, 30th day after surgery. The body composition was no significant difference between the two groups on the 7th day postoperatively. However, the loss of body composition such as skeletal muscle, body water, intracellular water, protein level and body cell volume were less in the study group compared with the control group on the 30th day after surgery. The incidence of postoperative complications in the prehabilitation group, ICU monitoring time, and hospitalization costs were significantly less than those in the control group. Conclusion: Prehabilitation intervention can obviously improve the nutritional status and the prognosis of the patients with esophageal cancer, and reduce the loss degree of their body composition.

Effect of Jin's Three-needle Acupuncture Combined with Occupational Therapy on Function of Upper Limbs for Stroke Patients with Hemiplegia / 中国康复理论与实践

Objective To explore the effects of Jin's Three-needle Acupuncture combined with occupational therapy on function of up-per limbs for stroke patients with hemiplegia. Methods 90 patients with stroke were randomly divided into control group (n=30), observa-tion group (n=30) and experiment group (n=30). The control group received routine rehabilitation, the observation group received occupa-tional therapy in addition, and the experiment group received Jin's Three-needle Acupuncture and occupational therapy in addition. They were assessed with Fugl-Meyer Assessment (FMA) of upper limbs and Barthel index (BI) before and 4 weeks after treatment. Results The scores of FMA and BI improved in all the groups after treatment (P<0.001). The difference of scores of FMA and BI before and after treat-ment were more in the experiment group than in the observation group (P<0.001), observation group than the control group (P<0.001). Con-clusion Jin's Three-needle Acupuncture combined with occupational therapy can further improve the motor function of upper limbs and ac-tivities of daily living for stroke patients with hemiplegia.

Effect of Jin's Three-needle Acupuncture Combined with Occupational Therapy on Function of Upper Limbs for Stroke Patients with Hemiplegia / 中国康复理论与实践

@#Objective To explore the effects of Jin's Three-needle Acupuncture combined with occupational therapy on function of upper limbs for stroke patients with hemiplegia. Methods 90 patients with stroke were randomly divided into control group (n=30), observation group (n=30) and experiment group (n=30). The control group received routine rehabilitation, the observation group received occupational therapy in addition, and the experiment group received Jin's Three-needle Acupuncture and occupational therapy in addition. They were assessed with Fugl-Meyer Assessment (FMA) of upper limbs and Barthel index (BI) before and 4 weeks after treatment. Results The scores of FMA and BI improved in all the groups after treatment (P<0.001). The difference of scores of FMA and BI before and after treatment were more in the experiment group than in the observation group (P<0.001), observation group than the control group (P<0.001). Conclusion Jin's Three-needle Acupuncture combined with occupational therapy can further improve the motor function of upper limbs and activities of daily living for stroke patients with hemiplegia.

Dietary restriction suppresses tumor growth, reduces angiogenesis, and improves tumor microenvironment in human non-small-cell lung cancer xenografts.

Non-small-cell lung cancer (NSCLC) is the leading cause of cancer deaths worldwide; however, only limited therapeutic treatments are available. The aim of present study was to elucidate the therapeutic effect of dietary restriction in human NSCLC xenografts. Adult female nude mice were injected subcutaneously in the right dorsal flank with NSCLC cell line A549 cells. 5 days after tumor implantation, animals were randomly divided into ad libitum-fed group (AL, 95% of average diary intake) or dietary-restriction-fed group (DR, 70% average diary intake). 24 days after implantation, it was found that DR inhibited tumor growth marked by lower tumor volume and weight. DR suppressed tumor proliferation marked by reduced proliferating cell nuclear antigen (PCNA) expression and activated mitochondria-mediated apoptosis. DR decreased microvessel density marked by decreased CD31 immunostaining and promoted vessel maturation marked by increased alpha-smooth muscle actin (α-SMA) and reduced Factor VIII expression. DR reduced intratumoral interstitial fluid pressure and attenuated tumor hypoxia detected by EF5 immunostaining. In addition, DR suppressed NFκB signaling pathway and downregulated its downstream proteins expression including cyclooxygenase 2 (COX-2) and inducible nitric oxide synthase (iNOS). DR suppressed phosphoinositide 3-kinase (PI3K)/AKT signaling pathway. In conclusion, dietary restriction suppresses tumor growth, reduces angiogenesis, and improves tumor microenvironment in human non-small-cell lung cancer xenografts. Dietary restriction could thus be envisaged as a nutritional countermeasure against non-small-cell lung cancer.