Totally mechanical Collard versus circular stapled cervical esophagogastric anastomosis for minimally invasive esophagectomy.

BACKGROUND: Previous studies have proposed that the totally mechanical Collard (TMC) method may reduce anastomotic leakage and stricture. This study aimed to compare the TMC method and the circular stapled (CS) method for cervical anastomosis after minimally invasive esophagectomy (MIE) for esophageal cancer. METHODS: From May 2017 to September 2020, 308 patients (165 in the CS group and 143 in the TMC group) were included in this study. The primary endpoints were anastomotic leakage and anastomotic stricture within 12 months. Propensity score matching was used to control potential selection bias. RESULTS: Anastomotic leak, anastomotic stricture, and refractory stricture (≥ 3 dilations) occurred in 30 (9.7%), 28 (9.1%), and 18 (5.8%) patients, respectively. The rate of anastomotic leak was similar in the CS and TMC methods (9.7 vs. 9.8%; P = 0.978), but anastomotic stricture (3.5 vs. 13.9%; P = 0.001) and refractory stricture (2.8 vs. 9.1%, P = 0.022) occurred less frequently in the TMC method. Propensity score matching yielded 128 patient pairs and confirmed these results. Multivariable analyses found that CS method, anastomotic leakage, and diabetes were independent predictors for both anastomotic stricture and refractory stricture. Subgroup analysis revealed that for patients with anastomotic leakage, the postoperative hospital stay in the TMC group was significantly longer than that in the CS group. CONCLUSION: In cervical anastomosis after MIE, the TMC method is superior to the CS method regarding anastomotic stricture and refractory stricture formation. However, compared to the CS method, the TMC method cannot lower the probability of anastomotic leakage, and anastomotic leakage with the TMC method requires a longer healing time.

Extensive open lobectomy experience is not a prerequisite for learning uniportal video-assisted thoracic surgery lobectomy.

BACKGROUND AND OBJECTIVES: Uniportal video-assisted thoracoscopic surgery (U-VATS) lobectomy has been increasingly adopted to manage early stage lung cancer. However, little information is available on whether this technique can be taught to surgeons inexperienced in open lobectomy. This study aimed to investigate the learning curve for U-VATS lobectomy performed by a single surgeon with limited open lobectomy experience. METHODS: From July 2018 to September 2020, 103 patients received U-VATS lobectomy for lung cancer by a single surgeon. The learning curve was assessed using three statistical methods: the moving average analysis, cumulative sum (CUSUM) analysis, and risk-adjusted CUSUM (RA-CUSUM) analysis. RESULTS: The moving average analysis showed a continuous decrease in operative time throughout the study period. The CUSUM analysis demonstrated three well-differentiated learning phases: Phase 1 (the initial 34 cases) representing the initial learning, Phase 2 (the middle 33 cases) representing the improvement of competence, and Phase 3 (the final 36 cases) representing technical proficiency. RA-CUSUM analysis revealed that the maximized cumulative surgical failure, defined as the maximum cumulative difference between the observed and predicted surgical failures, was found in the 61st case. CONCLUSIONS: U-VATS lobectomy is feasible for surgeons with limited open lobectomy experience. Multidimensional statistical analyses suggested that 61-67 cases were required to gain technical proficiency and ensure acceptable surgical outcomes.

Computed tomography-guided cyanoacrylate injection for localization of multiple ipsilateral lung nodules.

OBJECTIVES: To retrospectively assess the clinical effectiveness of CT-guided cyanoacrylate localization for multiple ipsilateral small pulmonary nodules (SPNs) and to determine the independent predictors for pneumothorax. METHODS: In total, 81 patients with 169 lesions undergoing CT-guided cyanoacrylate localization for multiple ipsilateral SPNs between September 2016 and July 2020 were enrolled (group M). Another 284 patients who received single SPN localization during the same period served as the control group (group S). Propensity score analysis was performed to minimize selection bias. Possible independent predictors for pneumothorax were evaluated using multivariate logistic regression analysis. RESULTS: Multiple ipsilateral SPN localization was successfully performed in all 81 patients. The incidences of successful targeting during localization and surgery were 100% and 98.8%, respectively. Seventy-seven patients (95.1%) underwent the procedure on the day before the surgery. Propensity matching created 81 pairs of patients. There were no significant differences in the incidence of successful targeting during localization and surgery, localization-related pain score, and additional morphine use between the two groups. However, group M was associated with a significant longer localization procedural time (p < 0.001) and a higher incidence of pneumothorax (p < 0.001). In multivariate analysis, position change was significantly associated with a sevenfold increase in the risk for pneumothorax (p = 0.001). CONCLUSIONS: CT-guided cyanoacrylate injection for multiple ipsilateral SPN localization was safe and reliable, and allowed a flexible surgical schedule, despite a lengthy procedure and an increased incidence of pneumothorax. Avoiding position change may help to reduce the occurrence of pneumothorax. KEY POINTS: • Compared to single SPN localization, multiple ipsilateral SPN localization using cyanoacrylate injection achieved comparable safety, reliability, and comfort. • CT-guided cyanoacrylate localization for multiple ipsilateral SPNs allowed a flexible surgical schedule. • Position change was the only independent risk factor for pneumothorax during the multiple ipsilateral SPN localization.

21- versus 25-mm Circular Staplers for Cervical Anastomosis: A Propensity-Matched Study.

BACKGROUND: The use of a small circular stapler (CS) has been reported to increase the incidence of benign anastomotic stricture of the intrathoracic anastomosis after esophagectomy, but no study has evaluated the effects of the CS size on cervical esophagogastrostomy. Based on a propensity-matched comparison, the present study was designed to determine whether the perioperative outcomes differ between 21- and 25-mm CSs after minimally invasive esophagectomy with cervical anastomosis. METHODS: From January 2015 to December 2017, 162 patients who received CS cervical esophagogastric anastomosis after minimally invasive esophagectomy for esophageal cancer were identified from our surgical database. A propensity-matched analysis was used to compare the outcomes between the 21- and 25-mm CS groups. Endpoints included anastomotic leak, dysphagia, reflux, stricture, and other major postoperative outcomes within 6 postoperative months. RESULTS: There were 69 and 93 patients in the 21- and 25-mm CS groups, respectively. Propensity matching produced 57 patients in each group. The two groups were not remarkably different in benign anastomotic stricture rate (P = 0.528). All strictures were resolved by balloon dilatation. The 25-mm CS group had a significantly longer operative time in cervical anastomosis than the 21-mm group (P = 0.005). No statistically significant differences in anastomotic leak rates, dysphagia scores, reflux scores, or other postoperative complications were noted between the two groups. CONCLUSIONS: The use of a 21-mm CS in minimally invasive esophagectomy with cervical esophagogastric anastomosis did not result in greater anastomotic stricture as compared with a 25-mm CS. The 21-mm CS was associated with a significantly shorter operative time.

Comparison of cyanoacrylate and hookwire for localizing small pulmonary nodules: A propensity-matched cohort study.

BACKGROUND: Localizing small pulmonary nodules (SPNs) is a challenge during thoracoscopic resection, but preoperative computed tomography (CT)-guided localization using either cyanoacrylate or hookwire can be helpful. This study compared the safety, efficiency, and operability of the two techniques. METHODS: From September 2013 to November 2018, 269 patients (269 SPNs) who underwent preoperative CT-guided SPN localization were enrolled. A propensity-matched analysis, incorporating 13 variables, was performed to control potential selection bias. RESULTS: All the patients were divided into two groups: CT-guided cyanoacrylate localization group (Group C, n = 149) and CT-guided hookwire localization group (Group H, n = 120). Eighty-six patients were propensity-matched in each group. All SPNs were successfully removed thoracoscopically, and no conversion was required. Localization-related complications in the two groups were similar, including intrapulmonary focal hemorrhage (p = 0.823), pneumothorax (p = 1.000), or hemoptysis (p = 0.121). For pain assessment and management, the cyanoacrylate localization saw a lower pain score (p < 0.001) and less morphine use (p < 0.001). In Group H, the localization took a significantly longer time (p < 0.001). Covering only the patients in Group C, the sub-analysis found that cyanoacrylate localization on the day before surgery did not compromise the accuracy of intraoperative targeting or increase the incidence of complications, compared with the localization on the day of surgery (all p > 0.05). CONCLUSION: Compared to hookwire localization, CT-guided cyanoacrylate localization decreased pain and morphine use and allowed flexible surgical schedules, suggestive of its preferability for the resection of SPNs.

Therapeutic Efficacy and Safety of Paeoniae Radix Rubra Formulae in Relieving Hyperbilirubinemia Induced by Viral Hepatitis: A Meta-Analysis.

OBJECTIVE: Hyperbilirubinemia is one of the most devastating pathologies induced by various liver diseases. Formulae related to Paeoniae Radix Rubra (PRR) at high doses have been applied to treat hyperbilirubinemia in traditional Chinese medicine (TCM). The aim of this systematic review and meta-analysis is to assess the efficacy and safety of formulae relevant to high-dose PRR in patients suffering from hyperbilirubinemia induced by viral hepatitis. METHODS: We performed a meta-analysis of randomized-controlled clinical trials to evaluate the efficacy and safety of formulae that apply a high dose of PRR for hyperbilirubinemia. Seven databases were searched until April, 2015. All studies were included according to detailed criteria and assessed for methodological quality. The outcome measurements were recorded for further analysis using the RevMan 5.2.11 software. RESULTS: Fifteen articles involving 1323 patients with hyperbilirubinemia were included. Formulae with high-dose PRR might promote the efficacy of either a combined application ([OR: 3.98, 95% CI (2.91, 5.43)]; P < 0.01) or a single application ([OR: 4.00, 95% CI (1.50, 10.68)]; P < 0.01) for hyperbilirubinemia. The indices of TBIL, ALT, and AST significantly decreased ([MD: -75.57, 95% CI (-94.88, -56.26)], [MD: -26.54, 95% CI (-36.19, -16.88)], and ([MD: -28.94, 95% CI (-46.26, -11.61)]; P < 0.01), respectively. In addition, formulae with high-dose PRR could enhance the treatment efficacy of hyperbilirubinemia triggered by hepatitis B ([OR: 2.98, 95% CI (1.75, 5.05)]; P < 0.01). Furthermore, the efficacy was enhanced with an increasing dosage of PRR. Two articles reported that no side effects occurred in clinical trials, and three studies noted that patients presented light digestive tract symptoms. CONCLUSION: Formulae relevant to high-dose PRR ameliorate hyperbilirubinemia and might constitute a promising therapeutic approach. For widespread acceptance by practitioners, more rigorously designed multicenter, double-blind, randomized, and large-scale controlled trials are required.

Paeoniflorin alleviates liver fibrosis by inhibiting HIF-1α through mTOR-dependent pathway.

HIF-1α/mTOR signaling pathway is considered to play a crucial role in genesis and progress of tissue fibrosis. The elevation of HIF-1α and mTOR is relevant to CCl4 induced liver fibrotic rats. Paeoniflorin has been consistently shown to exhibit multiple pharmacological effects in liver disease. However, so far, no research demonstrates the relationship between paeoniflorin and HIF-1α/mTOR fibrogenesis pathway in liver fibrosis. In this study, the liver fibrosis was performed by CCl4 rats and HSC-T6 cell line. The data demonstrated that paeoniflorin treatment could attenuate liver fibrosis and inhibit the activation of HSC. Moreover, paeoniflorin significantly enhanced hepatic function by decreasing serum level of ALT, AST and ALP, and increasing level of ALB, TP. Meanwhile, ECM degradation was modulated by paeoniflorin treated rats with a remarkable reduce of α-SMA and collagen III mRNA expression. Moreover, the alleviation effect of liver fibrosis was relevant to inhibiting HIF-1α and phosphor-mTOR. Our data indicate that paeoniflorin alleviates liver fibrosis by inhibiting HIF-1α expression partly through mTOR pathway and paeoniflorin may be a potential therapeutic agent for liver fibrosis.

Inhibition of calpain reduces oxidative stress and attenuates endothelial dysfunction in diabetes.

AIMS: The present study was to investigate the role of calpain in reactive oxygen species (ROS) production in endothelial cells and endothelium-dependent vascular dysfunction under experimental conditions of diabetes. METHODS AND RESULTS: Exposure to high glucose activated calpain, induced apoptosis and reduced nitric oxide (NO) production without changing eNOS protein expression, its phosphorylation and dimers formation in primary human umbilical vein endothelial cells (HUVECs). These effects of high glucose correlated with intracellular ROS production and mitochondrial superoxide generation. Selectively scavenging mitochondrial superoxide increased NO production in high glucose-stimulated HUVECs. Inhibition of calpain using over-expression of calpastatin or pharmacological calpain inhibitor prevented high glucose-induced ROS production, mitochondrial superoxide generation and apoptosis, which were concurrent with an elevation of NO production in HUVECs. In mouse models of streptozotocin-induced type-1 diabetes and OVE26 type-1 diabetic mice, calpain activation correlated with an increase in ROS production and peroxynitrite formation in aortas. Transgenic over-expression of calpastatin reduced ROS production and peroxynitrite formation in diabetic mice. In parallel, diabetes-induced reduction of endothelium-dependent relaxation in aortic ring was reversed by over-expression of calpastatin in mouse models of diabetes. However, the protective effect of calpastatin on endothelium-dependent relaxation was abrogated by eNOS deletion in diabetic mice. CONCLUSIONS: This study suggests that calpain may play a role in vascular endothelial cell ROS production and endothelium-dependent dysfunction in diabetes. Thus, calpain may be an important therapeutic target to overcome diabetes-induced vascular dysfunction.

Microcalorimetry coupled with chemometric techniques for toxicity evaluation of Radix Aconiti Lateralis Preparata (Fuzi) and its processed products on Escherichia coli.

As a widely used traditional Chinese medicine (TCM), Radix Aconiti Lateralis Preparata (Fuzi) is not only efficacious but also poisonous. Its toxicity and processed products should be taken into account and effectively evaluated. In this study, a non-invasive and non-destructive microcalorimetric method was employed to evaluate and compare the toxicity of Fuzi and its three processed products including Yanfupian, Heifupian, and Danfupian on Escherichia coli (E. coli). Some important metabolic information, such as the power-time curves and some quantitative thermokinetic parameters including growth rate constant k, heat output power P, inhibitory ratio I, and half inhibitory concentration IC50, of E. coli growth affected by various concentrations of Fuzi and its processed products were obtained. Combined with chemometric techniques including multivariate analysis of variance and principal component analysis on this information, it could be seen that Fuzi and its processed products could be distinguished according to their toxic effects on E. coli. The IC50 values of 14.6 mg/mL for Fuzi, 59.2 mg/mL for Yanfupian, 118.3 mg/mL for Heifupian, and 182.7 mg/mL for Danfupian illustrated that the sequence of toxicity on E. coli was Fuzi > Yanfupian > Heifupian > Danfupian. This study provided a useful method and idea of the combination of microcalorimetry and chemometrics for studying the toxic effects of TCMs and other substances.

Paeoniflorin protects against ANIT-induced cholestasis by ameliorating oxidative stress in rats.

Paeoniflorin, one of the primary bioactive components in Chi shao, are widely used in traditional Chinese medicine. A lot of evidences suggest that Paeoniflorin has potential anti-oxidant effects. However, whether Paeoniflorin plays roles in cholestasis is unclear. In this study, we examined the protective effect of Paeoniforin against alpha-naphthylisothicaynate (ANIT)-induced cholestasis in rats. Our data demonstrated that the high (0.2 g/kg body weight) and medium (0.1 g/kg body weight) doses of Paeoniflorin significantly prevented ANIT-induced changes in bile flow and the serum levels of total bilirubin, direct bilirubin, total bile acid, γ-glutamyltranspeptidase, glutamate-pyruvate transaminase, glutamate-oxaloacetic transaminase and alkaline phosphatase. Moreover, we also found that Paeoniflorin significantly inhibited nitric oxide and malondialdehyde production, and restored glutathione decrease induced by ANIT. EPR data further indicated that Paeoniflorin inhibited ANIT-mediated reactive oxygen species (ROS) production. The overexpression of NADPH oxidase 4 induced by ANIT were significantly reversed when treated with Paeoniflorin, suggesting that Paeoniflorin could scavenge ROS via inhibiting NADPH oxidase 4 expression. Paeoniflorin treatment could also relieve ANIT-induced liver pathological injuries as indicated by histological assay. These findings indicate that Paeoniflorin exerts a dose-dependent protective effect on ANIT-induced cholestatic liver injury in rats, and the mechanism of this activity is related to its attenuation of oxidative stress in liver tissue.

Establishment of a bioassay for the toxicity evaluation and quality control of Aconitum herbs.

Currently, no bioassay is available for evaluating the toxicity of Aconitum herbs, which are well known for their lethal cardiotoxicity and neurotoxicity. In this study, we established a bioassay to evaluate the toxicity of Aconitum herbs. Test sample and standard solutions were administered to rats by intravenous infusion to determine their minimum lethal doses (MLD). Toxic potency was calculated by comparing the MLD. The experimental conditions of the method were optimized and standardized to ensure the precision and reliability of the bioassay. The application of the standardized bioassay was then tested by analyzing 18 samples of Aconitum herbs. Additionally, three major toxic alkaloids (aconitine, mesaconitine, and hypaconitine) in Aconitum herbs were analyzed using a liquid chromatographic method, which is the current method of choice for evaluating the toxicity of Aconitum herbs. We found that for all Aconitum herbs, the total toxicity of the extract was greater than the toxicity of the three alkaloids. Therefore, these three alkaloids failed to account for the total toxicity of Aconitum herbs. Compared with individual chemical analysis methods, the chief advantage of the bioassay is that it characterizes the total toxicity of Aconitum herbs. An incorrect toxicity evaluation caused by quantitative analysis of the three alkaloids might be effectively avoided by performing this bioassay. This study revealed that the bioassay is a powerful method for the safety assessment of Aconitum herbs.

Rhein protects against acetaminophen-induced hepatic and renal toxicity.

This study investigated the possible protective effects and mechanism of rhein on Acetaminophen (APAP)-induced hepatotoxicity and nephrotoxicity in rats. Treatment of rats with APAP resulted in severe liver and kidney injuries, as demonstrated by drastic elevation of serum glutamate-pyruvate transaminase (GPT), glutamate-oxaloacetic transaminase (GOT), total bilirubin (TBIL), creatinine (CREA), urea nitrogen (UREA) levels and typical histopathological changes including necrosis, phlogocyte infiltration and fatty degeneration in liver, tubules epithelium swelling and severe vacuolar degeneration in kidney. APAP caused oxidative stress, as evidenced by increased reactive oxygen species (ROS) production, nitric oxide (NO) and malondiadehyde (MDA) levels, together with depleted glutathione (GSH) concentration in the liver and kidney of rats. However, rhein can attenuate APAP-induced hepatotoxicity and nephrotoxicity in a dose-dependent manner. Our results showed that GPT, GOT, UREA and CREA levels and ROS production were reduced dramatically, NO, MDA, GSH contents were restored remarkedly by rhein administration, as compared to the APAP alone treated rats. Moreover, the histopathological damage of liver and kidney were also significantly ameliorated by rhein treatment. These findings suggested that the protective effects of rhein against APAP-induced liver and kidney injuries might result from the amelioration of APAP-induced oxidative stress.

Disruption of Rac1 signaling reduces ischemia-reperfusion injury in the diabetic heart by inhibiting calpain.

Diabetes increases myocardial ischemia/reperfusion (I/R) injury. However, the underlying mechanisms remain incompletely understood. This study investigated the role of Rac1 signaling and calpain in exacerbated I/R injury in diabetic hearts. Mice with cardiac-specific deletion of Rac1 (Rac1-ko) and transgenic mice with cardiac-specific superoxide dismutase-2 (SOD2) or calpastatin overexpression were rendered diabetic with streptozotocin. Isolated perfused hearts were subjected to global I/R. After I/R, Rac1 activity was significantly enhanced in diabetic compared with nondiabetic hearts. Diabetic hearts displayed more severe I/R injury than nondiabetic hearts, as evidenced by more lactate dehydrogenase release and apoptosis and decreased cardiac function. These adverse impacts of diabetes were abrogated in Rac1-ko hearts or by perfusion with the Rac1 inhibitor NSC23766. In an in vivo I/R mouse model, infarct size was much smaller in diabetic Rac1-ko compared with wild-type mice. Inhibition of Rac1 signaling prevented NADPH oxidase activation, reactive oxygen species production, and protein carbonyl accumulation, leading to inhibition of calpain activation. Furthermore, SOD2 or calpastatin overexpression significantly reduced I/R injury in diabetic hearts and improved cardiac function after I/R. In summary, Rac1 activation increases I/R injury in diabetic hearts and the role of Rac1 signaling is mediated, at least in part, through calpain activation.

Histone deacetylase-3 activation promotes tumor necrosis factor-alpha (TNF-alpha) expression in cardiomyocytes during lipopolysaccharide stimulation.

Lipopolysaccharides (LPS) induce tumor necrosis factor-alpha (TNF-alpha) production in cardiomyocytes, which contributes to myocardial depression during sepsis. However, the underlying mechanisms remain not fully understood. This study was undertaken to investigate the contribution of histone deacetylase (HDAC) to TNF-alpha expression in cardiomyocytes and the signaling mechanism of LPS-induced HDAC activation. Here, we show for the first time that LPS increases HDAC activity and that inhibition of HDAC decreases LPS-stimulated TNF-alpha expression via the accumulation of NF-kappaB/p65 at the TNF-alpha promoter in cardiomyocytes. Using a positive screen, we have further identified HDAC3 as a specific member of the HDAC family able to regulate TNF-alpha production. Furthermore, our data reveal that LPS-induced HDAC activity is mediated through reactive oxygen species from mitochondria and c-Src signaling. In summary, this study demonstrates a novel signaling mechanism by which LPS via mitochondrial reactive oxygen species/c-Src/HDAC3 pathways mediate TNF-alpha expression in cardiomyocytes.

Differences in effects of Zuojin Pills and its similar formulas on Wei cold model in rats.

OBJECTIVE: To explore the effects of Zuojin Pills and its similar formulas on the stomach cold syndrome in a Wei cold model in rats. METHODS: The rat Wei cold model was established by intragastric administration of glacial NaOH, and the gastric mucosa injury indices, together with the levels of motilin and gastrin in the stomach, were determined. The preventive and curative effects of Zuojin Pills and its similar formulas on gastric mucosa injury were investigated. RESULTS: Zuojin Pills and its similar formulas could protect the gastric mucosa in the gastric cold model in rats at different levels. Fanzuojin Pills had the best effect in inhibiting gastric mucosa injury. CONCLUSION: The different pharmacological effects of Zuojin Pills and its similar formulas in the rat gastric cold model were partially correlated with the degrees in cold and heat properties of the formulas.

Rac1 is required for cardiomyocyte apoptosis during hyperglycemia.

OBJECTIVE: Hyperglycemia induces reactive oxygen species (ROS) and apoptosis in cardiomyocytes, which contributes to diabetic cardiomyopathy. The present study was to investigate the role of Rac1 in ROS production and cardiomyocyte apoptosis during hyperglycemia. RESEARCH DESIGN AND METHODS: Mice with cardiomyocyte-specific Rac1 knockout (Rac1-ko) were generated. Hyperglycemia was induced in Rac1-ko mice and their wild-type littermates by injection of streptozotocin (STZ). In cultured adult rat cardiomyocytes, apoptosis was induced by high glucose. RESULTS: The results showed a mouse model of STZ-induced diabetes, 7 days of hyperglycemia-upregulated Rac1 and NADPH oxidase activation, elevated ROS production, and induced apoptosis in the heart. These effects of hyperglycemia were significantly decreased in Rac1-ko mice or wild-type mice treated with apocynin. Interestingly, deficiency of Rac1 or apocynin treatment significantly reduced hyperglycemia-induced mitochondrial ROS production in the heart. Deficiency of Rac1 also attenuated myocardial dysfunction after 2 months of STZ injection. In cultured cardiomyocytes, high glucose upregulated Rac1 and NADPH oxidase activity and induced apoptotic cell death, which were blocked by overexpression of a dominant negative mutant of Rac1, knockdown of gp91(phox) or p47(phox), or NADPH oxidase inhibitor. In type 2 diabetic db/db mice, administration of Rac1 inhibitor, NSC23766, significantly inhibited NADPH oxidase activity and apoptosis and slightly improved myocardial function. CONCLUSIONS: Rac1 is pivotal in hyperglycemia-induced apoptosis in cardiomyocytes. The role of Rac1 is mediated through NADPH oxidase activation and associated with mitochondrial ROS generation. Our study suggests that Rac1 may serve as a potential therapeutic target for cardiac complications of diabetes.

Rac1 mediates sex difference in cardiac tumor necrosis factor-alpha expression via NADPH oxidase-ERK1/2/p38 MAPK pathway in endotoxemia.

The purpose of this study was to investigate the role of Rac1 and estrogen in sex difference of cardiac tumor necrosis factor-alpha (TNF-alpha) expression during endotoxemia. Endotoxemia was induced in male and female mice by peritoneal injection of lipopolysaccharide (LPS, 4 mg/kg). Compared with female mice, male mice produced more TNF-alpha in the heart 4 h after LPS treatment, which were correlated with higher Rac1 and NADPH oxidase activity, more phosphorylation of ERK1/2 and p38 MAPK, and up-regulation of toll-like receptor-4 (TLR-4) expression in male mice. Cardiac specific Rac1 knockout or administration of 17beta-estradiol down-regulated Rac1 expression, attenuated gp91(phox)-NADPH oxidase expression and activity, decreased phosphorylation of ERK1/2/p38 MAPK and inhibited cardiac TNF-alpha expression induced by LPS, suggesting an important role of Rac1 and estrogen in LPS-stimulated TNF-alpha expression in the heart. More importantly, the sex difference in TNF-alpha expression was abrogated by Rac1 knockout or gp91(phox) knockout and by administration of apocynin or N-acetylcysteine in LPS-stimulated mice. To investigate the functional significance of sex difference in endotoxemia, heart function was measured in isolated hearts with a Langendorff system. Male mice exhibited worse myocardial dysfunction compared with female in endotoxemia. Treatment of male mice with 17beta-estradiol attenuated myocardial dysfunction during endotoxemia. In conclusion, LPS induces Rac1 activation, which contributes to NADPH oxidase activity and phosphorylation of ERK1/2/p38 MAPK, leading to TNF-alpha expression in the heart. The sex difference in TNF-alpha expression is estrogen-dependent and mediated via Rac1 dependent NADPH oxidase/ERK1/2 and p38 MAPK pathway in LPS-stimulated hearts.

Investigations of free anthraquinones from rhubarb against alpha-naphthylisothiocyanate-induced cholestatic liver injury in rats.

The protective effects of anthraquinones from Rhubarb, a Chinese herbal medicine, consisting of the root and rhizome of Rheum palmatum L., R. tanguticum Maxim. Ex Balf., or R. officinale Baill. (family Polygonaceae) were investigated and compared in rats with liver injury induced by alpha-naphthylisothiocyanate. alpha-Naphthylisothiocyanate was given intragastrically in rats, liver injury with cholestasis developed within 36 hrs, as indicated by characteristic serum levels of glutamate-pyruvate transaminase, glutamic oxaloacetic transaminase, total bilirubin, direct bilirubin, alkaline phosphatase, gamma-glutamyltransferase and total bile acid. The intragastrical administration of rhein, aloe-emodin and physione to alpha-naphthylisothiocyanate-treated rats reduced significantly the serum level of both glutamate-pyruvate transaminase, glutamic oxaloacetic transaminase and the serum total bilirubin, direct bilirubin, alkaline phosphatase, gamma-glutamyltransferase and total bile acid. For all hepatic biochemical markers and cholestasis index, rhein was most efficient. By comparison, the administration of emodin and chrysophanol did not reduce the serum levels of hepatic enzymes glutamate-pyruvate transaminase and glutamic oxaloacetic transaminase but decreased the levels of serum total bilirubin, direct bilirubin, alkaline phosphatase, gamma-glutamyltransferase, and total bile acid, showing their partial protective effects on cholestatic liver injury. The liver in alpha-naphthylisothiocyanate-treated rats showed cholangiolitic hepatitis characterized by intrahepatic cholestasis, necrosis of hepatocytes and biliary epithelial cells and bile obstruction. The concurrent intragastrical administration of rhein reduced the severity of all morphological alteration, especially the neutrophil infiltration and sinusoid congestion. Rhein, aloe-emodin, and physione all exhibited protective effects on hepatocytes and cholangiocytes against alpha-naphthylisothiocyanate-induced damage, whereas emodin and chrystophanol provided partial protection.

Over-expression of calpastatin inhibits calpain activation and attenuates myocardial dysfunction during endotoxaemia.

AIMS: Lipopolysaccharide (LPS) induces cardiomyocyte caspase-3 activation and proinflammatory factors, in particular tumour necrosis factor-alpha (TNF-alpha) production, both of which contribute to myocardial dysfunction during sepsis. The present study was to investigate the roles of calpain/calpastatin system in cardiomyocyte caspase-3 activation, TNF-alpha expression, and myocardial dysfunction during LPS stimulation. METHODS AND RESULTS: In cultured adult rat cardiomyocytes, LPS (1 microg/mL) induced calpain and caspase-3 activity, and up-regulated TNF-alpha expression. These effects of LPS were abrogated by over-expression of calpastatin, an endogenous calpain inhibitor, transfection of calpain-1 siRNA, or various pharmacological calpain inhibitors. Furthermore, blocking gp91(phox)-NADPH oxidase prevented calpain and caspase-3 activation and decreased TNF-alpha expression in LPS-stimulated cardiomyocytes. To investigate the role of calpastatin in endotoxaemia, transgenic mice with calpastatin over-expression (CAST-Tg) and wild-type mice were treated with LPS (4 mg/kg, i.p.) or saline in the presence of calpain inhibitor-III (10 mg/kg, i.p.) for 4 h, and their heart function was measured with a Langendorff system. Over-expression of calpastatin significantly attenuated myocardial dysfunction (P < 0.05). Consistently, calpain activity, caspase-3 activity, and TNF-alpha expression were also reduced in CAST-Tg and calpain inhibitor-III compared with wild-type and vehicle-treated hearts, respectively. CONCLUSION: gp91(phox)-NADPH oxidase-mediated calpain-1 activation induces caspase-3 activation and TNF-alpha expression in cardiomyocytes during LPS stimulation. Over-expression of calpastatin inhibits calpain activation and improves myocardial function in endotoxaemia. The present study suggests that targeting calpain/calpastatin system may be a potential therapeutic intervention for septic hearts.

Effect of Radix isatidis polysaccharides on immunological function and expression of immune related cytokines in mice.

OBJECTIVE: To investigate the effect of Radix Isatidis polysaccharides (RIP) on the immunological function and expression of immune related cytokines in mice. METHODS: Sixty mice were randomly divided into six groups, the normal group, cyclophosphamide (Cy) model group, Levamisole (positive control) group, RIP low, medium and high dose groups (0.08 g/kg, 0.16 g/kg, 0.32 g/kg, respectively), ten in each group. By detecting the value of abdominal macrophage phagocytic index, serum hemolysin (HC50), proliferation of lymphocytes and expression of related cytokines, interleukin (IL-2) and interferon gamma (INF-gamma), the effect of RIP on immunological function and its mechanism were studied. RESULTS: RIP could improve the level of hemolysin in immunological function depressed mice. The results showed that the value of macrophage phagocytic index in the high dose RIP group increased from 1.11+/-0.13 to 1.42+/-0.26. The level of IL-2 and INF-gamma could be decreased by Cy to 38.12+/-6.88 ng/L and 139.23+/-29.87 ng/L, respectively, while RIP in high dose could increase the secretion of IL-2 and INF-gamma to 53.54+/-14.43 ng/L and 189.91+/-32.63 ng/L, respectively. CONCLUSION: RIP could enhance non-specific immunological function, humoral immunity and cellular immunity in mice.