Heme oxygenase­1 inhibits renal tubular epithelial cell pyroptosis by regulating mitochondrial function through PINK1.

Endotoxin-induced acute kidney injury (AKI) is commonly observed in clinical practice. Renal tubular epithelial cell (RTEC) pyroptosis is one of the main factors leading to the development of endotoxin-induced AKI. Mitochondrial dysfunction can lead to pyroptosis. However, the biological pathways involved in the potential lipopolysaccharide (LPS)-induced pyroptosis of RTECs, notably those associated with mitochondrial dysfunction, are poorly understood. Previous studies have demonstrated that heme oxygenase (HO)-1 confers cell protection via the induction of PTEN-induced putative kinase 1 (PINK1) expression through PTEN to regulate mitochondrial fusion/fission during endotoxin-induced AKI in vivo. Therefore, the present study investigated the role of HO-1/PINK1 in maintaining mitochondrial function and inhibiting the pyroptosis of RTECs exposed to LPS. Primary cultures of RTECs were obtained from wild-type (WT) and PINK1-knockout (PINK1KO) rats. An in vitro model of endotoxin-associated RTEC injury was established following treatment of the cells with LPS. The WT RTECs were divided into the control, LPS, Znpp + LPS and Hemin + LPS groups, and the PINK1KO RTECs were divided into the control, LPS and Hemin + LPS groups. RTECs were exposed to LPS for 6 h to assess cell viability, inflammation, pyroptosis and mitochondrial function. In the LPS-treated RTECs, the mRNA and protein expression levels of HO-1 and PINK1 were upregulated. Cell viability, adenosine triphosphate (ATP) levels and the mitochondrial oxygen consumption rate were decreased, whereas the inflammatory response, pyroptosis and mitochondrial reactive oxygen species (ROS) levels were increased. The cell inflammatory response and the induction of pyroptosis were inhibited, whereas the levels of mitochondrial ROS were decreased. In addition, the cell viability and ATP levels were increased in the WT RTECs following the upregulation of HO-1 expression. These effects were reversed by the downregulation of HO-1 expression. However, no statistically significant differences were noted between the LPS and the Hemin + LPS groups in the PINK1KO RTECs. Collectively, the findings of the present study indicate that HO-1 inhibits inflammation and regulates mitochondrial function by inhibiting the pyroptosis of LPS-exposed RTECs via PINK1.

Comparison of the supraglottic airway device BlockBusterTM and laryngeal mask airway Supreme in anaesthetised, paralyzed adult patients: a multicenter randomized controlled trial.

BACKGROUND: This multicenter prospective, randomized controlled clinical trial compared the clinical performance of supraglottic airway device (SAD) BlockBusterTM and laryngeal mask airway (LMA) Supreme for airway maintenance in anesthetized, paralyzed adult patients. METHODS: A total of 651 adult patients scheduled for elective surgery in 13 hospitals were randomly allocated into BlockBuster group (n = 351) or Supreme group (n = 300). The primary outcome was oropharyngeal leak pressure (OLP). Duration and ease of insertion, fiberscopic view of positioning, airway manipulations, and complications were also assessed. RESULTS: The OLP was significantly higher in BlockBuster group compared with Supreme group (29.9 ± 4.2 cmH2O vs 27.4 ± 4.3 cmH2O, p < 0.001). Success rate of insertion at the first attempt (90.2% vs 85.1%, p = 0.027), rate of optimal fiberscopic view (p = 0.002) and satisfactory positioning of SAD (p < 0.001) were significantly increased in BlockBuster group. CONCLUSIONS: Both SAD BlockBusterTM and LMA Supreme are safe, effective, and easy-to-use devices for airway maintenance in anesthetized, paralyzed adult patients, but the SAD BlockBusterTM is superior to LMA Supreme in terms of OLP, success rate at the first attempt, and fiber-optic view of positioning. TRIAL REGISTRATION: The trial is registered at www.chictr.org.cn (ChiCTR-ONC-16009105).

Analysis of clinical prognosis in patients with non-hepatic hyperammonemia.

ABSTRACT: The aim of this study was to evaluate the association of non-hepatic hyperammonemia (NHH) with the prognosis of critically ill patients with NHH.According to the serum ammonia level, the patients with NHH (n = 498) were retrieved by us. The risk factors of the mortality with NHH patients were investigated by conducting univariate and multivariate logistic regression analyses. A nomogram to predict the risk of hospital mortality was constructed. Receiver operating characteristic curve (ROC) analysis was conducted to compare nomogram (ammonia into a prognostic model, P1) with the simplified acute physiology II (SAPSII) and quick sequential organ failure assessment (qSOFA).Five independent factors for the mortality in patients with NHH were identified, including age, platelets, bun, hemoglobin, and ammonia. Models P1 using ammonia showed good prediction power. The AUROC of P1 (AUROC, 0.755 [95% CI, 0.713-0.796]) was higher than that of qSOFA (AUROC, 0.500 [95% CI, 0.449-0.551]), and SAPS II (AUROC, 0.703[95% CI, 0.658-0.748]).Ammonia was an independent prognostic predictor of mortality for NHH patients. We developed a nomogram that can predict hospital mortality with patients. Nomogram had superior discriminative power to qSOFA and SAPS II, indicating that the nomogram may have clinical utility.

HO-1/PINK1 Regulated Mitochondrial Fusion/Fission to Inhibit Pyroptosis and Attenuate Septic Acute Kidney Injury.

BACKGROUND: Endotoxin-associated acute kidney injury (AKI), a disease characterized by marked oxidative stress and inflammation disease, is a major cause of mortality in critically ill patients. Mitochondrial fission and pyroptosis often occur in AKI. However, the underlying biological pathways involved in endotoxin AKI remain poorly understood, especially those related to mitochondrial dynamics equilibrium disregulation and pyroptosis. Previous studies suggest that heme oxygenase- (HO-) 1 confers cytoprotection against AKI during endotoxic shock, and PTEN-induced putative kinase 1 (PINK1) takes part in mitochondrial dysfunction. Thus, in this study, we examine the roles of HO-1/PINK1 in maintaining the dynamic process of mitochondrial fusion/fission to inhibit pyroptosis and mitigate acute kidney injury in rats exposed to endotoxin. METHODS: An endotoxin-associated AKI model induced by lipopolysaccharide (LPS) was used in our study. Wild-type (WT) rats and PINK1 knockout (PINK1KO) rats, respectively, were divided into four groups: the control, LPS, Znpp+LPS, and Hemin+LPS groups. Rats were sacrificed 6 h after intraperitoneal injecting LPS to assess renal function, oxidative stress, and inflammation by plasma. Mitochondrial dynamics, morphology, and pyroptosis were evaluated by histological examinations. RESULTS: In the rats with LPS-induced endotoxemia, the expression of HO-1 and PINK1 were upregulated at both mRNA and protein levels. These rats also exhibited inflammatory response, oxidative stress, mitochondrial fission, pyroptosis, and decreased renal function. After upregulating HO-1 in normal rats, pyroptosis was inhibited; mitochondrial fission and inflammatory response to oxidative stress were decreased; and the renal function was improved. The effects were reversed by adding Znpp (a type of HO-1 inhibitor). Finally, after PINK1 knockout, there is no statistical difference in the LPS-treated group and Hemin or Znpp pretreated group. CONCLUSIONS: HO-1 inhibits inflammation response and oxidative stress and regulates mitochondria fusion/fission to inhibit pyroptosis, which can alleviate endotoxin-induced AKI by PINK1.

The PI3K-AKT-mTOR pathway activates recovery from general anesthesia.

We investigated roles of PI3K-AKT-mTOR pathway in recovery from general anesthesia. Sprague-Dawley rats divided into five groups: saline+artificial cerebrospinal fluid (ACSF; Group A), ketamine+ACSF (Group B), ketamine+IGF-1 (Group C), ketamine+PI3K inhibitor (Group D), and PI3K/Akt agonists (Group E). Proportion of δ waves on ECoGs was recorded. Rats were tested for duration of loss of righting reflex (LORR), ataxic period and behavior in Morris water maze. mRNA and protein expression of members of PI3K-AKT-mTOR pathway were measured by RT-qPCR and Western blots. Histopathologic changes in hippocampal tissues observed by HE staining. We found that the proportion of δ waves decreased in Group C, while increased in Group D compared with Group B; the durations of LORR and ataxic period were shorter in Group C, but longer in Group D. In Morris water maze, escape latency (EL) and duration and frequency of staying on platform was shorter in Group C and longer in Group D than in Group B. Group A exhibited low expression of proteins in PI3K-AKT-mTOR pathway, while p-AKT, p-mTOR and p-P70S6K expression increased in cerebral cortex, brain stem, and thalamus in Group C. By contrast, expression of those proteins was lower in Group D than Group B. Those proteins expressions were higher in Group E than in Group A. HE staining showed that anesthesia may induce cell apoptosis in rat hippocampal CA1 areas, and PI3K/Akt agonists could inhibit apoptosis. Our results suggest that activation of PI3K-AKT-mTOR pathway may promote recovery from general anesthesia and enhance spatial learning and memory.

[Mid-term follow-up results of laparoscopic radiofrequency ablation for renal cell carcinoma of T1aN0M0 stage].

OBJECTIVE: To determine the effect of laparoscopic radiofrequency ablation of T1aN0M0 renal cell carcinoma (RCC) with regular follow-up. METHODS: All patients underwent surgery from March 2006 to March 2009. Eight cases were solitary kidney. Twenty-two cases of left RCC and 18 cases of right RCC were diagnozed by ultrasonography and CT scanning.All of the cases were T1aN0M0 stage. No metastasis was found by iconography test. By ultrasound positioning, laparoscopic radiofrequency were performed on the renal tumor. All patients were followed up with eGFR and enhanced-CT. RESULTS: All patients underwent laparoscopic radiofrequency ablation surgery successfully. The mean operation time was (101 ± 19) minutes and the mean blood loss was (90 ± 14) ml (no blood transfusion pre- and post-operation). During postoperative follow-up, enhanced CT revealed complete ablation in 39 cases (the success rate was 97.5%), and 1 residue tumor was confirmed by enhanced CT 7 days post operation. This patient was under close surveillance because of solitary kidney. No progression of the residue tumor was found during the follow-up. One case of recurrence was confirmed by enhanced CT in 6 month after operation. The 3-year recurrence rate was 2.5%. No further intervation was performed on this patient and no change was found in the recurrence area during the follow-up. Both 3-year total survival rate and 3-year cancer specific survival rate were 100%. The mean eGFR was (72 ± 9) ml/(min·1.73 m(2)) in 3 years after surgery. There was no significant difference between pre-and post-operation (P > 0.05). CONCLUSION: Mid-term follow-up results show the effectiveness and safety of laparoscopic radiofrequency ablation in the treatment for T1aN0M0 RCC and have no negative influence on the renal function.

[Differential plasma protein profiles in patients with hyperlipidemia & atherosclerosis of different patterns of phlegm-stasis syndrome].

OBJECTIVE: To investigate the differential plasma protein profiles in patients with hyperlipidemia & atherosclerosis (H&A) of different patterns of phlegm-stasis syndrome (PSS) for seeking their biomarker proteins. METHODS: Two-dimensional gel electrophoresis and gel screening graphical analysis were performed on plasma proteins got from 146 patients; corresponding protein spots were fetched from the gel for two-stage mass-spectrometric analysis by quadruple time-of-flight mass spectrometry; then the differential proteins for PSS were discriminated by Fisher discriminate analysis. RESULTS: Excepting two uncertain proteins, 7 differential proteins were screened out from the 11 differentially expressed plasma protein spots with variability over 100% in the inter-block matching. Classic analysis found that haptoglobin precursor and fibrinogen gamma chain were possibly the plasma biomarker proteins for H & A; fibrinogen beta chain and apolipoprotein A-I precursor were that set apart PSS from non-PSS; fibrinogen gamma chain, albumin and apolipoprotein A-I precursor were for phlegm syndrome; haptoglobin precursor, adrenomedullin binding protein precursor, albumin and complement component C4 were for stasis syndrome; albumin and adrenomedullin binding protein precursor were for the phlegm-stasis mutual blocking syndrome. Moreover, the above mentioned expressions of possible marker proteins had their own special rule of changing in the transforming progress of PSS. CONCLUSION: This study reported, for the first time, the existence of evident variation of functional protein constitution in different patterns of PSS, and definite compatibility being detected in some functional proteins, which may be the marker proteins for making diagnosis and prognosis of PSS in H&A. Besides, preliminary proof for the transformation of PSS has gained at the functional protein level.

[Effects of apolipoprotein A1 and B gene polymorphism on avascular necrosis of the femoral head in Chinese population].

OBJECTIVE: To investigate the effects of Apo A1 and B gene polymorphism on avascular necrosis of the femoral head (ANFH) in north Chinese Han population. METHODS: Polymerase chain reaction-restriction fragment length polymorphisms (PCR-RFLP) technique was used in samples of 143 cases with documented ANFH and 92 healthy control matched by age and sex individuals selected from north Chinese Han nationality. The studied loci include promoter region (-75bp) and the intron 1 (+83 bp) of Apo A1 gene, Eco RI,Xba I of Apo B gene; polymerase chain reaction was used to study 3'-VNTR of Apo B gene. RESULTS: At -75 bp in promoter, the frequency of A/A genotype in ANFH group was significantly higher than that in control group (P < 0.01), while the frequency of G/A genotype in ANFH group was significantly lower than that in control group (P < 0.01). No difference was found in the frequency of genotype at +83bp in intron 1 of Apo A1 gene, Eco RI, Xba I and 3'-VNTR loci of Apo B gene. CONCLUSION: Apolipoprotein A1 gene A/A substitution at position -75 in promotor is associated with ANFH, the mutation may be one of the sensitive genes of ANFH, first reported inside and abroad. But no evident relationship was found between gene polymorphism of +75 bp loci of Apo A1 gene, Eco RI loci of Apo B gene, Xba I loci of Apo B gene or 3'-VNTR of Apo B gene and ANFH.

[Relationship between plasma protein expression profiles and states of Zang-Fu organs in patients with phlegm or blood stagnation syndromes due to hyperlipidemia and atherosclerosis].

OBJECTIVE: To investigate the relationship between the plasma biomarker proteins and the states of Zang-Fu organs in patients with phlegm or blood stagnation syndromes due to hyperlipidemia and atherosclerosis. METHODS: The states of Zang-Fu organs in 146 patients with hyperlipidemia and atherosclerosis were diagnosed by syndrome differentiation of traditional Chinese medicine. The plasma proteins from these patients were separated by two-dimensional polyacrylamide gel electrophoresis (2-DE). Differential protein profiling was established by Image Master 6.0 software, and the differential proteins were analyzed by quadrupole time of flight mass spectrometry (Q-TOF-MS). The association between the plasma biomarker proteins and the states of Zang-Fu organs was analyzed by graphical models. RESULTS: The biomarker proteins such as fibrinogen gamma chain, albumin and apolipoprotein AI (precursor) in discrimination of the patients with phlegm syndrome from phlegm accumulating with stagnation syndrome were correlated with the deficiency of kidney-qi, heart-qi and spleen-qi. Among the four biomarker proteins in discrimination of the patients with phlegm syndrome from blood stagnation syndrome, albumin, adrenomedullin binding protein (precursor) and haptoglobin (precursor) were correlated with the deficiency of kidney-qi and heart-qi, but complement component C4 was independent of the deficient Zang-Fu organs. The biomarker albumin was associated with the deficiency of kidney-qi, heart-qi and spleen-qi, and adrenomedullin binding protein (precursor) was correlated with the deficiency of spleen-qi in discrimination of the patients with blood stagnation syndrome from phlegm accumulating with stagnation syndrome. As the potential biomarker proteins in discrimination of the patients with non-phlegm and non-stagnation syndrome from phlegm accumulating with stagnation syndrome, the fibrinogen beta chain was related with the deficiency of kidney-qi, and apolipoprotein AI (precursor) was correlated with both the deficiency of kidney-qi and heart-qi. CONCLUSION: There exists inherent correlation between the states of Zang-Fu organs and the plasma probable biomarker proteins in the patients with different phlegm or blood stagnation syndromes due to hyperlipidemia and atherosclerosis.

[Applying matching matrix, factor analysis and clustering analysis to investigation of characteristics of syndromes of phlegm and blood stasis in patients with coronary heart disease].

OBJECTIVE: To investigate the characteristics of syndromes of phlegm and blood stasis in patients with coronary heart disease by multiple statistical methods of matching matrix, factor analysis and clustering analysis, and to provide some references for classification and normalization of diagnosis of syndromes of phlegm and blood stasis of coronary heart disease. METHODS: The correlations among 46 kinds of symptoms in syndrome of non-phlegm and non-blood stasis, syndrome of blood stasis, syndrome of phlegm and syndrome of phlegm-blood stasis blocking in 200 patients with coronary heart disease were analyzed by matching matrix, factor analysis and clustering analysis. RESULTS: The manifestations of tongue and pulse in syndromes of phlegm and blood stasis were significantly different from those in syndrome of non-phlegm and non-blood stasis. The pathogenesis of viscera in syndromes of phlegm and blood stasis lied in the heart and kidney, and the syndrome of deficiency of heart qi was the most common one while the syndrome of deficiency of kidney qi took the secondary place. The syndrome of phlegm was often accompanied by syndrome of deficiency of spleen qi. Only 15 ones of 46 clinical symptoms showed high frequency in concomitant appearance in syndrome of blood stasis, syndrome of phlegm and syndrome of phlegm-blood stasis blocking. Apart from having the common symptoms in syndrome of deficiency in origin, the syndrome of phlegm especially showed white and greasy fur and slippery pulse as well as distention and fullness of chest and abdominal distension; the syndrome of blood stasis showed purplish tongue and ecchymosis on tongue as well as fixed pain; and the syndrome of phlegm-blood stasis blocking showed the main symptoms of both syndrome of phlegm and syndrome of blood stasis. CONCLUSION: The statistical methods of matching matrix, factor analysis and clustering analysis are convenient, and can definitely indicate the clinical characteristics and syndrome differentiation of viscera of different syndromes of phlegm and blood stasis, which are beneficial to further research of diagnosis and differentiation of such syndromes of coronary heart disease.

[Study on relationship between polymorphism of apolipoprotein E gene and syndromes of phlegm and blood stasis in patients with coronary heart disease].

OBJECTIVE: To explore the relationship between polymorphism of apolipoprotein E (ApoE) exon 4 gene and different syndromes in patients with coronary heart disease (CHD). METHODS: Two hundred patients with CHD were divided into four groups according to syndrome differentiation, including syndrome of phlegm (PS), syndrome of blood stasis (BSS), syndrome of phlegm-blood stasis blocking (PBBS) and syndrome of non-phlegm and non-blood stasis (NPNBS). One hundred healthy volunteers were included in control group. Blood lipids were measured by routine examination. Total DNA of peripheral blood was extracted. ApoE genotypes were determined by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) technique. All data were analyzed by SAS software. RESULTS: (1)The occurrence rate of epsilon4 allele of ApoE in patients with CHD was 19.5%, significantly higher than 9.5% in the control group (P<0.05), and the E 3/4 genotype was especially more frequent (P<0.01). (2) The levels of total cholesterol (TC), total triglycerides (TG) and low-density lipoprotein cholesterol (LDL-C) in patients with epsilon4 were higher than those in patients without epsilon4 (P<0.01). (3) The frequencies of epsilon4 allele and E3/4 genotype in patients with PS were significantly higher than those in patients with BSS (P<0.05). CONCLUSION: ApoE epsilon4 allele, especially E3/4 genotype, is the risk factor of CHD. There is a relatively close relationship between patients with ApoE epsilon4 allele and PS. It may be one of the main susceptible genes in CHD patients with PS.