Therapeutic effect and autophagy regulation of myriocin in nonalcoholic steatohepatitis.

BACKGROUND: Ceramide plays pathogenic roles in nonalcoholic fatty liver disease (NAFLD) via multiple mechanisms, and as such inhibition of ceramide de novo synthesis in the liver may be of therapeutically beneficial in patients with NAFLD. In this study, we aimed to explore whether inhibition of ceramide signaling by myriocin is beneficial in animal model of NAFLD via regulating autophagy. METHODS: Sprague Dawley rats were randomly divided into three groups: standard chow (n = 10), high-fat diet (HFD) (n = 10) or HFD combined with oral administration of myriocin (0.3 mg/kg on alternate days for 8 weeks) (n = 10). Liver histology and autophagy function were measured. HepG2 cells were incubated with fatty acid with or without myriocin treatment. Lipid accumulation and autophagy markers in the HepG2 cells were analyzed. Serum ceramide changes were studied in 104 subjects consisting healthy adults, liver biopsy-proven patients with NAFLD and liver biopsy-proven patients with chronic hepatitis B (CHB). RESULTS: Myriocin reversed the elevated body weight and serum transaminases and alleviated dyslipidemia in HFD fed rats. Myriocin treatment significantly attenuated liver pathology including steatosis, lobular inflammation and ballooning. By qPCR analysis, it was revealed that myriocin corrected the expression pattern of fatty acid metabolism associated genes including Fabp1, Pparα, Cpt-1α and Acox-2. Further, myriocin also restored the impaired hepatic autophagy function in rats with HFD-induced NASH, and this has been verified in HepG2 cells. Among the sphingolipid species that we screened in lipidomic profiles, significantly increased ceramide was observed in NASH patients as compared to the controls and non-NASH patients, regardless of whether or not they have active CHB. CONCLUSIONS: Ceramide may play an important regulatory role in the autophagy function in the pathogenesis of NASH. Hence, blockade of ceramide signaling by myriocin may be of therapeutically beneficial in NASH. TRIAL REGISTRATION: Registration ID: ChiCTR-DDT-13003983 . Data of registration: 13 May, 2013, retrospectively registered.

Evidence that growth hormone can improve mitochondrial function in oocytes from aged mice.

Decline in successful conception decreases more rapidly after 38 years of age owing to follicular depletion and decreased oocyte quality. However, limited information is available regarding the underlying mechanism and the useful treatment. This study aimed to evaluate the effects of growth hormone supplementation on oocyte maturation in vivo in aged and young mice and to determine its effect on mitochondrial function. The influence of three different doses of recombinant human growth hormone (rhGH) (0.4, 0.8 and 1.6 mg/kg/day) for 8 weeks before ovarian stimulation was analyzed. Superovulated oocytes were released from the oviduct of 12-week-old and 40-week-old female C57BL/6J mice 14-16 h after administration of human chorionic gonadotropin. Ovarian follicle and morphological analysis and oocyte maturation parameters were then evaluated. This study is the first, to our knowledge, to report that medium- and high-dose rhGH significantly increases antral follicles in aged mice but anti-Müllerian hormone (AMH) levels. Furthermore, derived oocytes, MII-stage oocyte rate, ATP levels, mitochondrial membrane potential and frequencies of homogeneous mitochondrial distribution increased. In contrast, in both aged and young mice, the mtDNA copy numbers per oocyte were similar before rhGH administration, and upon saline administration, they did not differ significantly. We conclude that medium-dose rhGH supplementation before standard ovarian stimulation regimens improves oocyte quality in aged mice, probably by enhancing mitochondrial functionality.

Outcome of laparoscopy-guided hysteroscopic tubal catheterization for infertility due to proximal tubal obstruction.

STUDY OBJECTIVE: To determine pregnancy outcomes after laparoscopy-guided hysteroscopic tubal catheterization and to report its role in the era of in vitro fertilization. DESIGN: Clinical cases series (Canadian Task Force classification II-3). SETTING: Reproductive surgery center. PATIENTS: Patients with unilateral or bilateral proximal tubal obstruction as the only cause of infertility were included. INTERVENTIONS: Laparoscopy-guided hysteroscopic tubal catheterization. MEASUREMENTS AND MAIN RESULTS: Only the first spontaneous conception was considered. Cumulative conception rate (CCR) was calculated using Kaplan-Meier survival analysis. Of 168 women included, 107 (63.7%) had bilateral proximal obstruction and 61 (36.3%) had unilateral obstruction. The successful recanalization rate was 54.2% per tube and 61.9% per patient. In the 93 patients in whom at least 1 fallopian tube was successfully recanalized, 40 spontaneous pregnancies (43.0%) occurred within 24 months, of which 35 (37.6%) were intrauterine pregnancies and 28 (30.1%) resulted in live births. The CCR was 37.6% at 1 year and 43.7% at 2 years. Patients with unilateral obstruction in whom cannulation was successful had the highest CCR (60.7% at 2 years). CONCLUSION: Successful tubal cannulation led to significant improvement in the pregnancy rate, which suggests that women with a proximal tubal block could be considered for laparoscopy-guided hysteroscopic cannulation, which is still a viable alternative to in vitro fertilization.

[Related factors associated with pelvic adhesion and its influence on fallopian tube recanalization in infertile patients].

OBJECTIVE: To investigate factors with pelvic adhesions and the effect of different degrees pelvic adhesions on fallopian tube recanalization in infertile patients. METHODS: Total of 527 infertile patients undergoing hysteroscopy and laparoscopic surgery in Affiliated Hospital of Chinese People's Armed Police Forces Logistics College were studied retrospectively. According to the extent of pelvic adhesions, tubal umbrella adhesions and atresia, 377 cases were classified into adhesion groups, including 73 cases in grade I, 221 cases in grade II, 75 cases in grade III and 8 cases in grade IV based on adhesion score. The 150 cases with no obvious pelvic adhesion were matched as control group. Among 8 cases with grade IV ahesion were exluded from ahesion group the relationship between pelvic adhesions and related history, abdominal lesions, tubal patency and the prognosis were studied. RESULTS: (1) Related factors: the frequency of pelvic adhesion and more than 7 years of infertility of 23.9% (88/369) in adhesion group were significantly higher than 12.0% (18/150) in control groups. (2) HISTORY: compared with the control group (12.7%, 19/150; 28.7%, 43/150; 11.3%, 17/150; 12.0%, 18/150; 17.3%, 26/150), patients with pelvic adhesions present more incidence abortion (23.6%, 87/369), uterine cavity operation (38.2%, 141/369), ectopic pregnancy (20.9%, 77/369), pelvic inflammatory disease (25.5%, 94/369) and abdominopelvic surgery (31.4%, 116/369). (3) Endoscopy exploration: the incidence of hydrosalpinx (24.7%, 91/369), tube distorted (15.7%, 58/369) and salpingostomy (72.9%, 269/369) in adhesion group were higher than those in control group (2.0%, 3/150; 4.0%, 6/150; 12.0%, 18/150), but relatively lower incidence of pelvic endometriosis lesions (5.7%, 21/369) and mesosalpinx cysts (16.3%, 60/369) than those in control group (16.0%, 24/150; 30.0%, 45/150). The rate of proximal tubal recanalization (59.5%, 91/153) in adhesion group was lower than 75.4% (52/69) in control group. However, the rate of distant tubal recanalization of 84.4%, (281/333) in adhesion group and; 13/15 in control group didn't show statistical difference. (4) PROGNOSIS: the rate of ectopic pregnancy of 9.7% (29/299) in adhesion group was significantly higher than 3.1% (4/128) in control group. Among cases with grade III adhesion exhibited the highest rate of ectopic pregnancy (13.0%, 7/54; OR = 4.62, 95%CI: 1.29 - 16.50). (5) Multivariate analysis: it was found that more than two drug abortions (OR = 3.29, 95%CI: 1.34 - 8.07), pelvic and(or) abdominal surgery history (OR = 2.20, 95%CI: 1.35 - 3.57) and pelvic inflammatory disease history (OR = 1.54, 95%CI: 1.21 - 1.97) were risk factors with pelvic adhesions. CONCLUSION: More than or equal to two drug abortion history, pelvic inflammatory disease and pelvic and abdominal surgery damage were important factors for pelvic adhesions of infertility patients, which may decrease the possibility of proximal tubal recanalization and increase ectopic pregnancy risk.

Significance of platelet activating factor receptor expression in pancreatic tissues of rats with severe acute pancreatitis and effects of BN52021.

AIM: To investigate the dynamic changes and significance of platelet activating factor receptor (PAF-R) mRNA and protein in pancreatic tissues of rats with severe acute pancreatitis (SAP) and effects of BN52021 (Ginkgolide B). METHODS: Wistar male rats were randomly assigned to the negative control group (NC group), SAP model group (SAP group), and BN52051-remedy group (BN group), and each of the groups was divided into 6 subgroups at different time points after operation (1 h, 2 h, 3 h, 6 h, 12 h, and 24 h) (n = 10 in each). PT-PCR and Western blot methods were used to detect PAF-RmRNA and protein expression in pancreatic tissues of rats respectively. Pathological examination of pancreatic tissues was performed and the serum amylase change was detected. RESULTS: Serum amylase and pathological results showed the that SAP model was successfully prepared, BN52021 was able to decrease serum amylase, and the pathological ratings in BN group at 3 h, 6 h, and 12 h significantly decreased compared with those in the SAP group (8.85 +/- 0.39 vs 5.95 +/- 0.19, 9.15 +/- 0.55 vs 5.55 +/- 0.36, 10.10 +/- 0.65 vs 6.72 +/- 0.30, P < 0.05). The result of PAF-mRNA showed dynamic changes in SAP and BN groups, which increased gradually in early stage, reached a peak at 3 h (0.71 +/- 0.14 vs 0.54 +/- 0.14, 0.69 +/- 0.13 vs 0.59 +/- 0.04, P < 0.05), and decreased gradually later. There were significant differences at each time point except 1 h and 2 h, when compared with those in the NC group (0.71 +/- 0.14 or 0.69 +/- 0.13 vs 0.47 +/- 0.10, 0.38 +/- 0.08 or 0.59 +/- 0.04 vs 0.47 +/- 0.09, 0.25 +/- 0.07 or 0.29 +/- 0.05 vs 0.46 +/- 0.10, 0.20 +/- 0.06 or 0.20 +/- 0.04 vs 0.43 +/- 0.09, P < 0.05), whereas there was no significant difference between BN and SAP groups at each time point. The result of PAF-R protein showed that the change of PAF-R protein in the SAP group and the BN group was consistent with that of PAF-R mRNA. There were significant differences at each time point except 1 h, when compared with those in the NC group (0.90 +/- 0.02 or 0.80 +/- 0.05 vs 0.48 +/- 0.02, 1.69 +/- 0.06 or 1.58 +/- 0.02 vs 0.48 +/- 0.03, 1.12 +/-0.10 or 0.98 +/- 0.03 vs 0.49 +/- 0.09, 1.04 +/- 0.14 or 0.87 +/- 0.02 vs 0.52 +/- 0.08, 0.97 +/- 0.16 or 0.90 +/- 0.05 vs 0.49 +/- 0.10, P < 0.05), whereas there was no significant difference between the BN group and the SAP group. CONCLUSION: PAF-R plays an important role in occurrence and development of SAP. BN52021 exerts biological effects through competitively inhibiting the binding of increased both PAF and PAF-R expression rather than through decreasing PAF-R expression in pancreatic tissues.

Effect of BN52021 on NFkappa-Bp65 expression in pancreatic tissues of rats with severe acute pancreatitis.

AIM: To investigate dynamic changes and significance of expression of NF-kappaBp65 in pancreatic tissues of rats with severe acute pancreatitis (SAP), as well as BN52021 effects. METHODS: Wistar male rats were randomly divided into negative control group (NC group, n=60), SAP-model group (SAP group, n=60), and BN52021-treated group (BN group, n=60), and each of the above groups was respectively divided into 6 subgroups at different time points after operation (1 h, 2 h, 3 h, 6 h, 12 h, and 24 h) (n=10). By RT-PCR and Western blot, NF-kappaBp65 mRNA and its protein expression in pancreatic tissues of rats were detected respectively. RESULTS: The expression of NF-kappaBp65 mRNA dynamically changed in both SAP groups and BN groups. The mRNA level was higher in SAP groups than NC groups at 2 h, 3 h, 12 h, and 24 h after operation (P<0.05), higher in BN groups than NC groups at all time points (P<0.05), and higher in BN groups than SAP group at 1 h (P<0.05). The NF-kappaBp65 protein level was higher in SAP groups than NC groups at 1 h, 3 h, and 6 h (P<0.01), and 2 h, 12 h, and 24 h (P<0.05), higher in BN groups than NC groups at all time points (P<0.05), and lower in BN groups than SAP groups at 1 h, 3 h, and 6 h (P<0.05). CONCLUSION: The expression of NF-kappaBp65 in pancreatic tissues is dynamically changed and the changes play an important role in pathogenesis of SAP. BN52021 exerts therapeutic effects through reducing the expression level of NF-kappaBp65 protein in the early stage of SAP.

Serum Monounsaturated Triacylglycerol Predicts Steatohepatitis in Patients with Non-alcoholic Fatty Liver Disease and Chronic Hepatitis B.

Chronic liver disease is associated with lipid metabolic disruption. We carried out a study to determine serum lipidomic features of patients with non-alcoholic fatty liver disease (NAFLD) and active chronic hepatitis B (CHB) and explored the biomarkers for non-alcoholic steatohepatitis (NASH). Serum lipidomic profiles of healthy controls (n = 23) and of biopsy-proven NAFLD (n = 42), CHB with NAFLD (n = 22) and without NAFLD (n = 17) were analyzed by ultra-performance liquid chromatography-tandem mass spectrometry. There were distinct serum lipidome between groups of NAFLD and CHB without NAFLD. Most of the neutral lipids and ceramide were elevated in the NAFLD group but were decreased in the CHB without NAFLD group. Plasmalogens were decreased in both groups. Triacylglycerols (TAGs) with lower carbon numbers and double bonds were increased in subjects with NASH. Serum monounsaturated TAG was a significant predictor of NASH (OR = 3.215; 95%CI 1.663-6.331) and positively correlated with histological activity (r = 0.501; P < 0.001). It showed good predictability for NASH in the NAFLD group [area under the receiver operating characteristic curves (AUROC) = 0.831] and was validated in the CHB group (AUROC = 0.833); this characteristic was superior to that of cytokeratin-18 and alanine transaminase. The increase in monounsaturated TAG might be a specific marker for NASH in both NAFLD and CHB patients.

The 100 most cited articles in ectopic pregnancy: a bibliometric analysis.

Ectopic pregnancy (EP) remains a major gynecological emergency and is a cause of morbidity or even mortality in women. As a consequence, top citation analysis of EP research in database of the Science Citation Index Expanded is needed to assess the publication trends of leading countries/territories and institutes as well as the research hotspots of EP. A total of 4881 articles relevant to EP were retrieved in the database of the Science Citation Index Expanded from 1965 to present, in which the 100 top-cited articles were selected for further analysis. The number of citations ranged from 81 to 482 (131.57 ± 69.76), with a time span of 40 years between 1969 and 2009. These citation classics came from 14 countries, and 65 of the articles came from the United States. Yale University in Connecticut led the list of classics with six papers. The 100 top-cited articles were published in 32 journals, in which the journal of Fertility and Sterility published the most (23 papers). Stovall TG and Ling FW published the highest number of studies (6 papers each). Articles that originated in the United States and that were published in high-impact journals were most likely to be cited in the field of EP research. Bibliometric analysis was used to provide a historical perspective on the progress in EP research over the past 50 years. Citation analysis is a feasible tool to comprehensively recognize the advances of EP research in the past and future research.

G[alpha]i2 and G[alpha]q expression change in pancreatic tissues and BN52021 effects in rats with severe acute pancreatitis.

OBJECTIVE: To investigate change of G[alpha]i2, G[alpha]q mRNA, and their proteins in severe acute pancreatitis (SAP) and BN52021 effects. METHODS: Rats were assigned into negative-controlled group (NC), SAP-modeled group (SAP), and BN52051-remedial group (BN). Each group was assigned into 6 subgroups at different time points. G[alpha]i2, G[alpha]q mRNA, and their proteins were determined. RESULTS: In the SAP group, G[alpha]i2 at 12 and 24 hours and G[alpha]q at 1 and 6 hours were remarkably higher than those in the NC group; in the BN Group, G[alpha]i2 is not remarkably different from that in the SAP group, but G[alpha]q at 1 and 6 hours was lower than those in the SAP group (P < 0.01), and G[alpha]i2 at 12 hours was higher than that in the NC group (P < 0.05), but G[alpha]q was not remarkably different from that in the NC group; in the SAP group, G[alpha]i2 and G[alpha]q proteins were higher than those in the NC group (P < 0.05); in the BN group, G[alpha]i2 proteins at 6, 12, and 24 hours and G[alpha]q proteins were lower than those in the SAP group (P < 0.05), and G[alpha]i2 and G[alpha]q proteins at each time phase point except 24 hours were higher than those in the NC group (P < 0.05). CONCLUSIONS: G[alpha]i2, G[alpha]q mRNA, and their proteins in SAP increase. BN52021 decreases G[alpha]i2 and G[alpha]q.

Theoretical investigation of electron transfer transition in tetracyanoethylene-contained organic complexes.

In this work, the authors use complete active space self-consistent field method to investigate the photoinduced charge-separated states and the electron transfer transition in complexes ethylene-tetracyanoethylene and tetramethylethylene-tetracyanoethylene. Geometries of isolated tetracyanoethylene, ethylene, and tetramethylethylene have been optimized. The ground state and the low-lying excited states of ethylene and tetracyanoethylene have been optimized. The state energies in the gas phase have been obtained and compared with the experimentally observed values. The torsion barrier of tetracyanoethylene has been investigated through the state energy calculation at different conformations. Attention has been particularly paid to the charge-separated states and the electron transfer transition of complexes. The stacked conformations of the donor-acceptor complexes have been chosen for the optimization of the ground and low-lying excited states. Equilibrium solvation has been considered by means of conductor-like screening model both in water and in dichloromethane. It has been found that the donor and tetracyanoethylene remain neutral in complexes in ground state (1)A(1) and in lowest triplet state (3)B(1), but charge separation appears in excited singlet state (1)B(1). Through the correction of nonequilibrium solvation energy based on the spherical cavity approximation, pi-->pi* electron transfer transition energies have been obtained. Compared with the experimental measurements in dichloromethane, the theoretical results in the same solvent are found higher by about 0.5 eV.