[Effect of macrophage-derived exosomes on promoting hepatic stellate cell activation and platelet-derived growth factor expression].

Objective: To explore the effect of macrophage-derived exosomes on the activation of hepatic stellate cells and its possible mechanism. Methods: Differential ultracentrifugation was used to extract macrophage exosomes. The exosomes were co-cultured with the mouse hepatic stellate cell line JS1, and a control group was established with phosphate buffered saline (PBS). Cell immunofluorescence was used to observe the expressional conditions of F-actin. Cell counting kit-8 (CCK8) was used to detect the survival rate of JS1 cells in the two groups. The activation indices of JS1 cells [collagen type Ⅰ (Col Ⅰ) and α-smooth muscle actin (α-SMA)] and its key signal pathway activation index expression level [transforming growth factor (TGF)-ß1/Smads, platelet-derived growth factor (PDGF)] in the two groups were determined using Western blot and RT-PCR. Data comparison between two groups was performed using an independent sample t-test. Results: The membrane structure of exosomes was clearly observed by transmission electron microscopy. The expression of exosome marker proteins CD63 and CD81 was positive, suggesting that exosomes were successfully extracted. Exosomes were co-cultured with JS1 cells. Compared with the PBS control group, there was no statistically significant difference in the proliferation rate of JS1 cells in the exosomes group (P＞0.05). The expression of F-actin was significantly increased in the exosome group. The mRNA and protein expression levels of α-SMA and ColⅠwere significantly increased in exosome group JS1 cells (all P＜0.05). The mRNA relative expression levels of α-SMA in PBS and exosome group were 0.25±0.07 and 1.43±0.19, respectively, while that of ColⅠ was 1.03±0.04 and 1.57±0.06, respectively. The mRNA and protein expressions of PDGF were significantly increased in exosome group JS1 cells (P＜0.05). The mRNA relative expression levels of PDGF in the PBS group and exosome group were 0.27±0.04 and 1.65±0.12, respectively. There were no statistically significant differences in the mRNA and protein expressions of TGF-ß1, Smad2 and Smad3 between the two groups (P＞0.05). Conclusion: Macrophage-derived exosomes significantly promote the activation of hepatic stellate cells. JS1 cells may be the underlying mechanism for the up-regulation of PDGF expression.

[Development of multiple organ dysfunction syndrome in patients with Coronavirus Disease 2019: clinical characteristics and risk factors].

Objective: To explore the clinical characteristics and risk factors of patients with Coronavirus Disease 2019 (COVID-19) when developing multiple organ dysfunction syndrome (MODS). Methods: Data from 458 inpatients of confirmed COVID-19 in Wuhan, Shanghai and Tongling from December 29, 2019 to March 24, 2020 were retrospectively collected. COVID-19 was confirmed by real-time RT-PCR of throat swab samples. Data of demographics, clinical presentation, laboratory tests, imaging findings, treatment and prognosis were obtained from medical record and compared between COVID-19 patients with and without MODS. Risk factors for the development of MODS were analyzed by univariate and multivariate logistic regression analysis. Results: Of the 458 COVID-19 patients (266 from Wuhan, 208 from Shanghai, and 24 from Tongling), 103 developed transient or persistent MODS in the course. More male patients were found in those with MODS (72.8% vs 54.6%, P=0.001). And MODS patients were of older age (72.8% vs 54.6%, P=0.001), more chronic comorbidities (68.0% vs 43.4%, P<0.001), and longer onset-to-admission interval (9.0 vs 7.0 d, P<0.001). In addition, patients with MODS had more expectoration (45.6% vs 29.9%, P=0.003) and shortness of breath (52.4% vs 19.4%, P<0.001), dysfunction of various systems, decreased cellular immunity and elevated IL-6 (9.6 vs 7.6 g/L, P=0.015) in laboratory tests, isolation of other pathogens (18.4% vs 5.6%, P<0.001), and infiltration of all five lobes (75.3% vs 57.6%, P=0.003). During hospitalization, patients with MODS needed a higher proportion of comprehensive treatment and reached a mortality rate of 66.0%. Independents risk factors for development of MODS in COVID-19 patients were: onset-to-admission interval>7 days (OR=2.17, 95%CI: 1.11-4.22, P=0.023), shortness of breath (OR=3.19, 95%CI: 1.60-6.37, P=0.001), lymphocyte count<1×109/L (OR=2.67, 95%CI: 1.31-5.46, P=0.007), blood urea nitrogen>7mol/L (OR=6.27, 95%CI: 2.80-14.08, P<0.001), procalcitonin>0.1 ng/mL (OR=2.48, 95%CI: 1.20-5.13, P=0.014), and C-reactive protein>10 mg/L (OR=3.92, 95%CI: 1.41-10.89, P=0.009). Conclusions: COVID-19 patients with MODS were of higher severity and mortality. Early identification of high-risk groups with MODS according to risk factors may be helpful for early treatment.

The correlation between tissue Doppler imaging and N-terminal pro-brain natriuretic peptide concentration during left ventricular diastolic function recovery after coronary artery bypass grafting.

OBJECTIVE: This study aims to evaluate the changes in left ventricular diastolic function after coronary artery bypass grafting through tissue Doppler imaging (TDI) and N-terminal pro-brain natriuretic peptide (NT-proBNP) concentration and the correlation between the two. PATIENTS AND METHODS: A total of 133 patients who underwent coronary artery bypass grafting between January 2016 and December 2018 were included in this study. Echocardiography and NT-proBNP concentration were reviewed pre-operation, 1 month post-operation, and 3-6 months post-operation. The transmitral peak flow velocity (E) of the mitral valve was measured at each of the three-time points using spectral Doppler imaging. The mitral annulus displacement (Ea peak and Aa peak) was then measured at each of the time points using TDI, and the E/Ea ratio was calculated. Subsequently, the correlation of the E value, Ea value, and E/Ea ratio with NT-proBNP concentration was statistically analyzed. RESULTS: The data obtained at the three-time points were compared with the respective concentrations of NT-proBNP. Multiple linear regression analysis revealed a correlation between NT-proBNP concentration and E value, Ea value, and E/Ea ratio. CONCLUSIONS: Left ventricular diastolic function gradually recovered at 1 month and 3-6 months after coronary artery bypass grafting. There was a correlation between TDI-related values and NT-proBNP concentration.

Serum metabolic markers and metabolic pathways in rats with metabolomic cecal ligation and puncture-induced sepsis.

The aim of this study was to explore the dynamic changes in characteristic serum metabolic markers and pathways during early sepsis in rats. By using cecal ligation and puncture (CLP), we made rat models of sepsis, which were randomly divided into 5 groups with 10 rats in each group: group A, group B, group C, group D, and group E. We collected 2 mL of arterial blood at 0, 6, 12, 24, and 48 hours from rats in group A-E respectively and isolated serum via centrifugation. Next, adopting metabolomics analysis methods, we screened for metabolites from the animal serum with statistically and biologically significant abundance changes, and used the KEGG database to analyze the respective metabolic pathways. In all, our findings reveal that D-glucosamine 6-phosphate, D-glucosamine phosphate, α-D-glucosamine 1-phosphate, D-glucosamine 1-phosphate, and 5-hydroxy isocyanate decline continuously from 12 hours, while L-phenylalanine, (S) -α-amino-ß-phenylpropionic acid, 5-methoxy indole acetic acid salt, 5-methoxy indole acetic acid, goose deoxyglycolic acid salt, goose deoxyglycolic acid, and Chen's deoxygenated sugar alcohol started to decrease from 6 hours. Additionally, 3.2,3-Bis-O-(geranyl geranyl)-sn-glycerol- 1-phosphoric acid-L-serine levels rose continuously from 12 hours. We found 13 differentially regulated ions, primarily ones involved in pathways responsible for the metabolism of sugar, amino acids, and lipids, which are related to the disorder of energy metabolism. Our findings mark serum-derived D-glucosamine and its phosphorous derivatives as characteristic metabolic markers of sepsis in rats.

[Surgical outcome evaluation and prediction analysis of laparoscopic left sided hepatectomy for hepatolithiasis].

Objective: To explore the risk factors of long-term treatment outcomes and establish predicting model for laparoscopic left hepatectomy in hepatolithiasis. Methods: Clinical data of 108 patients with hepatolithiasis who underwent laparoscopic left sided hepatectomy and with complete follow-up data were retrospectively collected from June 2011 to June 2016 at the Second Affiliated Hospital of Nanchang University. Twenty-six males and 82 females were enrolled. The age was (52.4±11.7) years (range:20-80 years) , and the median follow-up time was 36 months (range: 24-83 months) . Patients were randomly divided into training group (79 cases) and validation group (29 cases) with a ratio of about 3∶1. Twenty-five preoperative and intraoperative clinical factors were selected for potential factors that might affect long-term outcomes, and quality of life was used as an surrogate evaluation index. Univariate analysis and multivariate logistic regression analysis were used to investigate the potential risk factors, and to construct and validate the predictive nomogram for surgical outcomes. Results: Among 108 patients, 10 patients (9.3%) had residual stones, 8 patients (7.4%) had recurrent stones, 12 patients (11.1%) had recurrent cholangitis and 3 patients (2.8%) died. Univariate analysis showed that history of hepatobiliary surgery, gender, activation of partial thromboplastin time, alkaline phosphatase, use of choledochoscopy, postoperative stone residual, serum creatinine, postoperative biliary drainage and operation time were risk factors that may affect long-term outcomes (all P<0.15) . Multivariate analysis showed that the history of previous hepatobiliary surgery (OR=2.305, 95% CI: 0.383-4.227, P=0.019) , postoperative biliary drainage (OR=2.043, 95% CI: 0.182-4.209, P=0.048) , operation time ≥262.5 minutes (OR=1.971, 95% CI: 0.154-4.023, P=0.045) were independent risk factor affecting long-term outcomes. Based on the above factors, the predictive nomogram model was constructed. Internal and external validations showed good discrimination (area under the curve of receiver operating curve>0.7) and calibration (Hosmer-Lemeshow test: P>0.05) performance, which indicated that the prediction effect was favorable. Conclusions: History of previous hepatobiliary surgery, postoperative biliary drainage and operation time ≥262.5 minutes are independent risk factors for long-term outcome. The predictive nomogram model based on risk factors relates to surgical outcomes presented good clinical predictive effects, which might contribute to the prediction of the long-term outcomes of laparoscopic left sided hepatectomy for hepatolithiasis.

Longitudinal Microstructural Changes in Traumatic Brain Injury in Rats: A Diffusional Kurtosis Imaging, Histology, and Behavior Study.

BACKGROUND AND PURPOSE: Traumatic brain injury is a major public health problem worldwide. Accurately evaluating the brain microstructural changes in traumatic brain injury is crucial for the treatment and prognosis assessment. This study aimed to assess the longitudinal brain microstructural changes in traumatic brain injury in the rat using diffusional kurtosis imaging. MATERIALS AND METHODS: Diffusional kurtosis imaging was performed in a group of 5 rats at preinjury and 3, 14, and 28 days after traumatic brain injury. The diffusional kurtosis imaging parameters were measured in the bilateral cortex, hippocampus, and corpus callosum. Another 4 groups of 5 rats were used in brain immunohistochemistry analysis of neuron (neuron-specific nuclear protein [NeuN]), astroglia (glial fibrillary acidic protein [GFAP]), microglia (ionized calcium binding adaptor molecule 1 [Iba-1]), and myelin (myelin basic protein [MBP]) in the same area as the diffusional kurtosis imaging parameter measurements. Furthermore, 2 groups of 6 rats underwent a Morris water maze test at 28 days after traumatic brain injury. The diffusional kurtosis imaging parameters, immunohistochemistry results, and Morris water maze test results were compared longitudinally or between traumatic brain injury and control groups. RESULTS: Compared with baseline, traumatic brain injury in the rat showed higher mean kurtosis and mean diffusivity values in the ipsilateral perilesional cortex and hippocampus and lower fractional anisotropy values in the corpus callosum (P < .05). The traumatic brain injury group showed higher staining of GFAP and Iba-1 and lower immunohistochemistry staining of NeuN and MBP in all ipsilateral ROIs (P < .05). There was no significant difference in the contralateral ROIs in diffusional kurtosis imaging parameters or immunohistochemistry results. The Morris water maze test revealed lower platform crossing times in the probe test (P < .05). CONCLUSIONS: Our study indicated that there were longitudinal changes in diffusional kurtosis imaging parameters, accompanied by multiple pathologic changes at different time points following traumatic brain injury, and that mean kurtosis is more sensitive to detect microstructural changes, especially in gray matter, than mean diffusivity and fractional anisotropy.

L-364,718 potentiates electroacupuncture analgesia through cck-a receptor of pain-related neurons in the nucleus parafascicularis.

Electroacupuncture (EA) has been successfully used to alleviate pain produced by various noxious stimulus. Cholecystokinin-8 (CCK-8) is a neuropeptide involved in the mediation of pain. We have previously shown that CCK-8 could antagonize the analgesic effects of EA on pain-excited neurons (PENs) and pain-inhibited neurons (PINs) in the nucleus parafascicularis (nPf). However, its mechanism of action is not clear. In the present study, we applied behavioral and neuroelectrophysiological methods to determine whether the mechanisms of CCK-8 antagonism to EA analgesia are mediated through the CCK-A receptors of PENs and PINs in the nPf of rats. We found that focusing radiant heat on the tail of rats caused a simultaneous increase in the evoked discharge of PENs or a decrease in the evoked discharge of PINs in the nPf and the tail-flick reflex. This showed that radiant heat could induce pain. EA stimulation at the bilateral ST 36 acupoints in rats for 15 min resulted in an inhibition of the electrical activity of PEN, potentiation of the electrical activity of PIN, and prolongation in tail-flick latency (TFL), i.e. EA stimulation produced an analgesic effect. The analgesic effect of EA was antagonized when CCK-8 was injected into the intracerebral ventricle of rats. The antagonistic effect of CCK-8 on EA analgesia was reversed by an injection of CCK-A receptor antagonist L-364,718 (100 ng/µl) into the nPf of rats. Our results suggest that the pain-related neurons in the nPf have an important role in mediating EA analgesia. L-364,718 potentiates EA analgesia through the CCK-A receptor of PENs and PINs in the nPf.

Energy dependence of restitution in the gastric mucosa.

Rapid epithelial repair (restitution) after injury is required to maintain barrier function of the gastrointestinal mucosa and skin and is thought to be a highly ATP-dependent process that would be inhibited under hypoxic conditions. However, little is known about the metabolic pathways required for restitution. Thus, this study was undertaken to evaluate, in vitro, the role of oxidative respiration and glycolysis in restitution after injury. To this end, restitution of the bullfrog gastric mucosa was evaluated under the following conditions: 1) blockade of mitochondrial respiration; 2) blockade of glycolysis; or 3) absence of glucose. The extent of mucosal repair after injury was evaluated by electrophysiology and morphology. Cell migration, repolarization, and the formation of tight junctions after injury occurred during blockade of mitochondrial respiration, whereas the recovery of mucosal barrier function did not. In contrast, glycolytic inhibition completely blocked all aspects of restitution by inhibiting the migration of surface epithelial cells. Restitution occurred in tissues incubated with glucose-free solutions, suggesting that cells contain sufficient glucose (glycogen) to drive glycolysis for many hours. Our results demonstrate that the glycolytic pathway is essential for restitution after injury in the bullfrog gastric mucosa and that all but complete repair of barrier function occurs in the absence of mitochondrial respiration.

Circulating levels of retinol, tocopherol and carotenoid in Nepali pregnant and postpartum women following long-term beta-carotene and vitamin A supplementation.

OBJECTIVE: To characterize circulating carotenoid and tocopherol levels in Nepali women during pregnancy and post-partum and to determine the effects of beta-carotene and vitamin A supplementation on their concentration in serum. DESIGN: Randomized community supplementation trial. SETTING: The study was carried out from 1994 to 1997 in the Southern, rural plains District of Sarlahi, Nepal. SUBJECTS: A total of 1431 married women had an ascertained pregnancy, of whom 1186 (83%) provided an analyzable serum sample during pregnancy; 1098 (77%) provided an analyzable 3-4 months post-partum serum sample. INTERVENTIONS: Women received a weekly dose of vitamin A (7000 microg RE), beta-carotene (42 mg) or placebo before, during and after pregnancy. Serum was analyzed for retinol, alpha-tocopherol, gamma-tocopherol, beta-carotene, alpha-carotene, lycopene, lutein + zeaxanthin, and beta-cryptoxanthin concentrations during mid-pregnancy and at approximately 3 months post-partum. RESULTS: Compared to placebo, serum retinol, beta-carotene, gamma-tocopherol, beta-cryptoxanthin and lutein + zeaxanthin concentrations were higher among beta-carotene recipients during pregnancy and, except for beta-cryptoxanthin, at postpartum. In the vitamin A group, serum retinol and beta-cryptoxanthin were higher during pregnancy, and retinol and gamma-tocopherol higher at postpartum. Lutein + zeaxanthin was the dominant carotenoid, regardless of treatment group, followed by serum beta-carotene. Serum lycopene level was lowest, and very low compared to the US population. Serum retinol was higher, and carotenoid and alpha-tocopherol lower, at postpartum than during pregnancy in all groups. CONCLUSIONS: Pregnant and lactating Nepali women have lower serum carotenoid and tocopherol levels than well-nourished populations. beta-carotene supplementation appeared to increase levels of tocopherol and other carotenoids in this population.

Simultaneous measurement of axial strain, temperature, and transverse load by a superstructure fiber grating.

A novel and simple fiber-optic sensor based on a superstructure fiber grating for simultaneous measurement of temperature, axial strain, and transverse load is proposed and demonstrated. By measurement of the shift and split of broadband and narrow-band loss peaks, one can determine the temperature, axial strain, and transverse load simultaneously over the ranges 0-140 degrees , 0-1200muepsilon, and 0-0.3 kg/mm, respectively.

Influence of prenatal iron and zinc supplements on supplemental iron absorption, red blood cell iron incorporation, and iron status in pregnant Peruvian women.

BACKGROUND: It is estimated that 60% of pregnant women worldwide are anemic. OBJECTIVE: We aimed to examine the influence of iron status on iron absorption during pregnancy by measuring supplemental iron absorption, red blood cell iron incorporation, and iron status in pregnant women. DESIGN: Subjects were 45 pregnant Peruvian women (33+/-1 wk gestation), of whom 28 received daily prenatal supplements containing 60 mg Fe and 250 microg folate without (Fe group, n = 14) or with (Fe+Zn group, n = 14) 15 mg Zn, which were were consumed from week 10 to 24 of gestation until delivery. The remaining 17 women (control) received no prenatal supplementation. Iron status indicators and isotopes were measured in maternal blood collected 2 wk postdosing with oral (57Fe) and intravenous (58Fe) stable iron isotopes. RESULTS: Maternal serum ferritin and folate concentrations were significantly influenced by supplementation (P < 0.05). Serum iron was also significantly higher in the Fe than in the Fe+Zn (P < 0.03) or control (P < 0.001) groups. However, the supplemented groups had significantly lower serum zinc concentrations than the control group (8.4+/-2.3 and 10.9+/-1.8 micromol/L, respectively, P < 0.01). Although percentage iron absorption was inversely related to maternal serum ferritin concentrations (P = 0.036), this effect was limited and percentage iron absorption did not differ significantly between groups. CONCLUSIONS: Because absorption of nonheme iron was not substantially greater in pregnant women with depleted iron reserves, prenatal iron supplementation is important for meeting iron requirements during pregnancy.

PIP: The influence of iron status on iron absorption during pregnancy was examined among pregnant Peruvian women. This was done by measuring supplemental iron absorption, red blood cell iron incorporation and iron status. The subjects were 45 pregnant Peruvian women (33 +or- 1 week gestation) who were divided into 2 groups. The first group of 28 pregnant women received daily prenatal supplements containing 60 mg of iron and 250 mcg of folate with or without 15 mg of zinc, from week 10 to 24 of gestation until delivery. The second group of 17 women served as the control group. The control group was not given prenatal supplementation. The iron status indicators and isotopes were measured in maternal blood collected 2 weeks postdosing with oral iron-57 and intravenous iron-58 stable isotopes. The results showed that supplementation significantly influenced the maternal serum ferritin and folate concentrations (P 0.05). The serum iron of the iron group was significantly higher than that of the iron + zinc group (P 0.03) or control group (P 0.001). However, the serum zinc concentrations were lower in the supplemented group than in the control group. Even though the percentage of iron absorption was inversely related to maternal serum ferritin concentration, the effect was limited and the percentage of iron absorption did not differ significantly between the two groups. Considering that the absorption of nonheme iron was not substantially greater in pregnant women with depleted iron reserves, it was concluded that prenatal iron supplementation is essential for meeting iron requirements, especially during pregnancy.

Antagonistic effect of CCK-8 on morphine-inhibited electrical and contractile activities of rat jejunum in vitro.

In the present investigation, antagonistic action of cholecystokinin octapeptide (CCK-8) against morphine on the electrical and contractile activity of rat jejunum in vitro was studied. The results showed that the potentiation of acetylcholine (ACh) on both the burst of spike and the contractility were inhibited by morphine, which could be completely antagonized by CCK-8. The CCK-8 effect, again, could be suppressed by CCK-A receptor antagonist devazepide (10 nmol/L), but partially by CCK-B receptor antagonist L-365, 260 at 10 nmol/L or completely at concentration of 30 nmol/L. The above results demonstrated that the antagonism of CCK-8 on morphine was mediated by both CCK-A and CCK-B receptors.

[Effect of tremulacin on actions of SRS-A and histamine].

The effect of tremulacin (TRC) extracted from Mao Bai Yang (Folia Populus tomentosa Carr) on actions of SRS-A and histamine were investigated by using isolated guinea pig ileum and spectrofluorometric assay. TRC was found to inhibit the contraction of isolated guinea pig ileum induced by histamine and SRS-A, in a dose-dependent manner with IC50 of 1.78 x 10(-4) mol.L-1 and 2.51 x 10(-4) mol.L-1, respectively. TRC at the dose of 10(-4) mol.L-1 inhibited SRS-A release from sensitized isolated guinea pig lung. While at the dose of 10(-5) mol.L-1 inhibited histamine release from the peritoneal mast cells in sensitized rats. These results indicate that inhibition of the release of histamine and SRS-A may play an important role in the mechanism of antiinflammatory actions of TRC.

Antiinflammatory effects of Tremulacin, a Salicin-related substance isolated from Populus tomentosa Carr. leaves.

Tremulacin was shown to inhibit carrageenan-induced paw edema in rats and croton oil-induced ear edema in mice. It was also found to inhibit peritoneal leucocyte migration in rats and acetic acid-induced writhing responses in mice. Experiments with isolated longitudinal muscle strips of sensitized guinea pig ileum showed that tremulacin decreased the biosynthesis of Slow Reaction Substance of Anaphylaxis. Tremulacin exerted inhibitory effects on leukotriene B4 biosynthesis in intrapleural leucocytes. These results suggest that the mechanism of antiinflammatory actions of tremulacin is relevant to inhibition of 5-lipoxygenase activity. This is quite different from non-steroid antiinflammatory drugs, such as aspirin, which inhibits prostaglandin synthesis and being a cyclooxygenase inhibitor.