Protective effects of gliclazide on high glucose and AGEs-induced damage of glomerular mesangial cells and renal tubular epithelial cells via inhibiting RAGE-p22phox-NF-kB pathway.

OBJECTIVE: Gliclazide is one of the most widely used therapeutic drugs for diabetes. As a second-generation sulfonylurea oral hypoglycemic drug, it can lower blood glucose level and delay the occurrence and development of diabetic nephropathy (DN). However, the underlying mechanism remains unclear. Therefore, the aim of this study was to explore whether gliclazide had protective effects on high glucose and advanced glycation end products (AGEs)-induced injury of human mesangial cells (HMCs) and renal tubular epithelial cells. MATERIALS AND METHODS: HMC and renal tubular epithelial cell lines [human kidney 2 (HK-2)] were cultured in vitro. All cells were then divided into the follow groups: 1) blank control group (5.6 mmol/L glucose), 2) AGEs group [400 µg/mL AGE-bovine serum albumin (AGE-BSA)], 3) high glucose group (25 mmol/L glucose), 4) gliclazide + AGEs group (400 µg/mL AGE-BSA + 20 µmol/L gliclazide) and 5) gliclazide + high glucose group (25 mmol/L glucose + 20 µmol/L gliclazide). Cell counting kit-8 (CCK-8) assay was adopted to determine cell viability. Flow cytometry was used to detect cell apoptosis. The levels of malondialdehyde (MDA), superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) were measured as well. Furthermore, the mRNA expressions of receptor for AGE (RAGE), p22phox and nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) were measured via fluorescence quantitative Real-time polymerase chain reaction (qRT-PCR). RESULTS: Compared with control group, significantly accelerated apoptosis of HMCs and HK-2, increased MDA level, decreased SOD and GSH-Px levels, and up-regulated mRNA expressions of RAGE, p22phox and NF-κB were observed in HMCs and HK-2 of high glucose group and AGEs group. Meanwhile, there were obviously alleviated apoptosis of HMCs and HK-2, decreased MDA level, increased SOD and GSH-Px levels, as well as down-regulated mRNA expressions of RAGE, p22phox and NF-κB in HMCs and HK-2 of gliclazide group compared with high glucose and AGEs group. Furthermore, significant correlations were found between the mRNA expression of RAGE and the apoptosis rate of HMCs and HK-2 (HMCs: r=0.701, p=0.004 and HK-2: r=0.633, p=0.011). CONCLUSIONS: Gliclazide has protective effects on high glucose and AGEs-induced damage of glomerular mesangial cells and renal tubular epithelial cells via inhibiting RAGE-NADPH oxidase-NF-kB pathway.

Immune efficacy of a porcine circovirus type 2 vaccine purified using Gram-positive enhancer matrix surface display technology.

AIMS: Purification of porcine circovirus type 2 (PCV2) using Gram-positive enhancer matrix (GEM) surface display technology and immunogenicity evaluation of the purified antigen. METHODS AND RESULTS: A recombinant bifunctional protein containing a protein anchor domain and a 'virus anchor' domain was designed as a protein linker (PL) between PCV2 and GEM particles. By incubating with PL and GEM particles sequentially, PCV2 could be purified and enriched through a simple centrifugation process with GEM surface display technology. Our data showed that one unit (2·5 × 109 particles) of GEM particles with 80 µg PL could purify 100 ml of PCV2-containing culture supernatant (viral titre: 106·5 TCID50 per ml-1 ) with a recovery rate up to 99·6%. The impurity removal efficiency of this method, calculated according to decreased total protein content during purification, was approximately 98%. Furthermore, in vivo experimentation showed that piglets immunized with purified PCV2 could elicit strong immune responses to prevent against PCV2 infection. CONCLUSION: Porcine circovirus type 2 could be efficiently purified and enriched with GEM display technology via a crucial PL, and the purified PCV2 could elicit effective immune responses against PCV2 infection. SIGNIFICANCE AND IMPACT OF THE STUDY: The GEM-based purification method established here is cost-efficient and high-throughput, and may represent a promising large-scale purification method for PCV2 vaccine production.

The shape change index (SCI) of inferior vena cava (IVC) measuring by transabdominal ultrasound to predict the presence of septic shock in intensive care unit (ICU) patients.

OBJECTIVE: This study is designed as prospective and observational research of patients with sepsis. It was carried out in the intensive care unit (ICU). We investigated the shape change index (SCI) of inferior vena cava (IVC) measured with trans-abdominal ultrasound to detect the signs of septic shock. The aim of this research was to find the most effective tool in predicting shock in patients compared with that of other parameters such as brain natriuretic peptide (BNP), lactate, variation index of inferior vena cava IVC-VI, and extravascular lung water index (EVLWI). PATIENTS AND METHODS: We suppose that SCI can be used as the safest and most sensitive tool in the early recognition of septic dysfunction. The observational study was conducted in the Department of ICU, Shandong Provincial Hospital Affiliated to Shandong University from January 2016 to December 2017. SCI of IVC, serum lactate, BNP, IVC-VI, and EVLWI concentrations were measured in 30 sepsis patients. All studied biomarkers were analyzed and contrasted according to the score of the Sequential Organ Failure Assessment (SOFA). Pearson correlation analysis was analyzed to statistic the relationship between variables. RESULTS: We showed the correlation of BNP value, lactic acid value, IVC-VI, EVLWI, and SCI of IVC in sick patients suffering septic shock. Positive correlation was observed in the BNP value, lactic acid value, IVC-VI, EVLWI, and SCI of IVC (r=0.447, p=0.013; r=0.484, p=0.007; r=0.423, p=0.023; r=0.638, p<0.001; r=0.599, p<0.001; respectively). However, the SCI and EVLWI showed a stronger correlation with the SOFA than the others. SCI of IVC, as estimated by transabdominal ultrasound, was more accurate than the other commonly used non-invasive predictors. EVLWI, as an accurate and classical predictor, was an invasive predictor. SCI of IVC was faster, more convenient and safer than the other. CONCLUSIONS: SCI of IVC was faster, more convenient and safer than the other commonly used non-invasive predictors. Early recognition and diagnosis of sepsis may improve patient outcome.

Relationship between the flavonoid composition and flower colour variation in Victoria.

Victoria (Nymphaeaceae), an annual or perennial aquatic plant genus, contains only two species: V. amazonica (Poepp.) J. C. Sowerby and V. cruziana A. D. Orb. Both species have large floating leaves and variable flower colour. Both Victoria species are night bloomers, which have white petals on the first blooming night that then turn pink or ruby red on the second blooming day. The mechanism of the colour change of Victoria petals during anthesis is still unclear. In this study, flavonoids in Victoria petals of both species were evaluated and quantified by high-performance liquid chromatography with photodiode array detection (HPLC-DAD) and by ultra-performance liquid chromatography coupled with tandem mass spectrometry (UPLC-MS/MS) for the first time. In total, 14 flavonoids were detected in Victoria petals, including 4 anthocyanins and 10 flavonols. The flavonoid compositions differed across the two species, resulting in different colours between the inner and outer petals. With increased anthocyanin content across blooming days, the colour of Victoria flowers changed over time. The results of this study will improve understanding of the chemical mechanism of colour formation and lay the foundation for selective colour breeding in Victoria.

Management of extensive retrohepatic vena cava defect in recipients of living donor liver transplantation.

Certain complexities, such as extensive vena caval injury, unexpected dense adhesions between liver and retrohepatic vena cava, and liver tumor abutting retrohepatic vena cava, sometimes warrant resection of vena cava during living-donor liver transplantation. Because the donor graft is devoid of vena cava, reconstruction of the retrohepatic cava is required, which can be done with the use of either a cryopreserved venous graft or an artificial conduit. With only a few published reports, the experience in vena cava reconstruction with the use of expanded polytetrafluoroethylene (ePTFE) during living-donor liver transplantation remains limited. We present our experience of 4 patients who successfully underwent vena caval resection during liver transplantation for various indications, which was subsequently reconstructed with the use of ePTFE grafts. All of these patients except 1 recovered well without any undue complications, such as thrombosis or outflow inadequacies, thus proving this extensive surgical treatment to be a successful and life-saving procedure, though meticulous skills are prerequisite.

Clinical relevance of sHLA-G-mediated with better graft acceptance in early posttransplantation.

The aim of this study was to measure the level of soluble human leukocyte antigen (sHLA-G) in renal transplant patients, to determine the relationship between these levels and the occurrence of acute rejection episodes, and to identify their influence on graft acceptance early posttransplantation. sHLA-G, as measured by an enzyme-linked immunosorbent assay, was significantly increased (P < .01) early posttransplantation (3 months); the other group maintained low levels throughout the study. The latter group displayed a high incidence of acute rejection episodes and a lower clearance of serum creatinine with a longer period for hemoglobin to recover to normal (P < .01). These results suggested that HLA-G participates in the induction of immunologic tolerance in these recipients.

A high model for end-stage liver disease score should not be considered a contraindication to living donor liver transplantation.

OBJECTIVE: To analyze the outcomes of patients with high Model for End-Stage Liver Disease (MELD) scores who underwent adult-to-adult live donor liver transplantation (A-A LDLT). MATERIALS AND METHODS: From September 2002 to October 2010, a total of 152 adult patients underwent A-A LDLT in our institution. Recipients were stratified into a low MELD score group (Group L; MELD score≤30) and a high MELD score group (Group H; MELD score>30) to compare short-term and long-term outcomes. RESULTS: Of the 152 adult patients who underwent A-A LDLT, 9 were excluded from the analysis because they received ABO-incompatible grafts. Group H comprised 23 and Group L 120 patients. The median follow-up was 21.5 months (range, 3 to 102 m). The mean MELD score was 15.6 in Group L and 36.7 in Group H. There were no significant differences in the mean length of stay in the intensive care unit (Group L: 3.01 days vs Group H: 3.09 days, P=.932) or mean length of hospital stay (Group L: 17.89 days vs. Group H: 19.91 days, P=0.409). There were no significant differences in 1-, 3-, or 5-year survivals between patients in Groups L versus H (91.5% vs 94.7%; 86.4% vs 94.7%; and 86.4% vs 94.7%; P=.3476, log rank). CONCLUSION: The short-term and long-term outcomes of patients with high MELD scores who underwent A-A LDLT were similar to those of patients with low MELD scores. Therefore, we suggest that high MELD scores are not a contraindication to LDLT.

Hepatic artery reconstruction in living donor liver transplantation: running suture under surgical loupes by cardiovascular surgeons in 180 recipients.

OBJECTIVES: The aim of our study was to retrospectively investigate the outcomes of hepatic artery (HA) reconstruction by cardiovascular surgeons in adult-to-adult living donor liver transplantation (A-A LDLT). METHODS: From April 2007 to April 2011, 187 recipients underwent A-A LDLT. After excluding seven ABO-incompatible transplant recipients, we reviewed the courses of 180 patients including 125 men and 55 women of mean age 52.5±9.2 years (range=23-71). One hundred seventy-seven patients received right-lobe grafts with inclusion of middle hepatic vein (MHV); two, right-lobe grafts without MHV; and one, left-lobe graft. A continuous, single-stitch, running suture with the parachute technique was used for HA reconstruction. The anastomosis was performed by cardiovascular surgeons employing surgical loupes with 4.5× magnification. RESULTS: The mean time for an arterial reconstruction was 10.7±4.0 minutes (median=10, range=4-30). Hepatic arterial thrombosis (HAT) was encountered in 3 (1.66%) patients. One HAT that developed on postoperative day 1 was successfully rescued by the intra-arterial infusion of urokinase. Another patient required reoperation due to a redundant kinked HA. A third HAT patient underwent successful retransplantation with a cadaveric graft on postoperative day 6. In our series, no delayed HAT was detected and no recipient deaths were related to HAT. CONCLUSION: HA reconstruction with a running suture under surgical loupes is a feasible technique in A-A LDLT. A speedy reconstruction can be performed by an experienced cardiovascular surgeon with a low incidence of HAT.

Living donor right hepatectomy with inclusion of the middle hepatic vein: outcome in 200 donors.

Venous congestion of segments V and VIII of the graft is observed frequently in right-lobe living donor liver transplants (LDLT) without middle hepatic vein (MHV) drainage. It can cause graft dysfunction and failure. Inclusion of the MHV in the right lobe graft allows optimal venous drainage but can pose adverse effects for the donor. From May 2005 to April 2011, we performed 202 right-lobe LDLTs using grafts that all (except two) contained the MHV. The mean duration of donor surgery was 558±132 minutes (median 540, range 332-1100), and estimated blood loss 441±309 mL (median 350, range 35-3200). No donor was admitted to the intensive care unit postoperatively. The mean hospital stay was 8.7±2.1 days (median 8, range 6-22). Postoperatively, 39 donors (19.5%) experienced Clavien grade I and II complications, mostly minor wound infections or massive ascites necessitating diuretic therapy. Seven (3.5%) donors displayed Clavien grade III complications, including five bile leakages requiring endoscopic retrograde biliary drainage and two abdominal wound dehiscences requiring repair under general anesthesia. There was no donor death. In conclusion, inclusion of the MHV in a right-lobe LDLT was safe for most donors.

Simultaneous liver and kidney transplantation from different living donors.

OBJECTIVES: To present our experience with simultaneous living donor liver and kidney (SLK) transplantation from two different living donors. PATIENTS AND METHODS: We performed five SLK transplantations from two different living donors from November 2006 to December 2010. Four patients were males and one, female. Their age range was 47 to 66 years (mean, 55 years). The primary liver diseases included hepatitis B virus (n=2), alcoholic liver cirrhosis (n=2), cryptogenic liver disease (n=1), and hepatitis C virus with hepatocellular carcinoma (n=1). All five patients had chronic renal failure: four were on hemodialysis (H/D) and one on chronic ambulatory peritoneal dialysis for 1 to 20 years. Liver implantation was performed first, followed by kidney transplantation. The liver and kidney teams worked closely to shorten the ischemia time. RESULTS: All surgical procedures were performed uneventfully and all recipients and donors survived the operations. Good liver graft function was noted in all five patients. The patient with both anti-T- and anti-B-cell positive crossmatch tests developed hyperacute rejection of the kidney graft requiring its immediate removal. This patient was maintained on regular H/D afterward. The other four patients displayed good renal function. No evidence of severe acute rejection was noted during the follow-up period (range, 9-55 months) among patients treated with tacrolimus-based immunosuppression. CONCLUSION: We suggest that SLK transplantation be performed with organs from two different instead of a single live donor.

Aperture-domain processing and its applications in ultrasound imaging: a review.

This paper provides a review of advanced algorithms for ultrasound image formation and signal processing that are based on aperture-domain data (i.e. the data recorded by individual channels prior to beam summation). First aperture-domain data are defined and their properties described, then two specific examples of phase-aberration correction and vector velocity estimation are presented. For phase-aberration correction, sidelobe-reduction techniques based on the coherence of the received aperture-domain data were tested with clinical breast data; the mean improvements in the contrast and contrast-to-noise ratios were 6.9 dB and 23.2 per cent, respectively. For flow estimation, a conventional scanner can only estimate the flow velocity parallel to the beam axis. The proposed flow estimation technique uses aperture-domain data for two-dimensional flow-velocity estimation. The experimental results demonstrate that the estimation errors for the proposed technique are 2.18 per cent and 18.11 per cent in the axial and lateral velocity components, respectively. Other applications in which aperture-domain data can be used are also discussed.

Intracavernosal pressure characteristics analyzed by power spectral density for identifying the phase of penile erection.

Knowledge of the time course of penile erection is very important to understanding erection physiology. The changes in the intracavernosal pressure (ICP) and the different phases of an erection are pivotal to the ability to produce and maintain a rigid penile erection. This study investigated an objective and low-invasiveness method for identifying different erection phases based on an innovative ICP analysis technique. Blood infuses into the corpora cavernosa and causes the ICP to increase. The ICP usually exhibits tiny oscillations at the frequency of the heartbeat when it increases from diastole to systole. The characteristic oscillation amplitudes corresponding to the period when the full and rigid erection phases begin can be extracted by power spectral density analysis. The reliability and accuracy of the proposed method was verified by the Bland-Altman graphs indicating a good agreement with the existing method that compares the ICP with the arterial pressure. Moreover, all of the intraclass correlation coefficient values were close to 1.00, with the lower limit of the 95% confidence interval exceeding 0.75. The described novel objective and low-invasiveness method can therefore be used for identifying the full and rigid erection phases of the penis in urological investigations during different erection phases.

Innovative technique to reconstruct two branches of the right hepatic artery in living donor liver transplantation.

Hepatic arterial thrombosis is a critical complication in living donor liver transplantation (LDLT). Two separate branches of the right hepatic artery (RHA) are sometimes observed and addressed by anastomosis of the larger branch first, then checking backflow from the smaller branch. If not good, the smaller branch must be reconstructed. We used the cystic artery as a conduit for the reconstruction. Meticulous dissection was performed to identify all branches of the hepatic artery in the donor operation. The length of cystic artery preserved was as long as possible. The cystic arterial stump was anastomosed to the stump of the posterior branch the of RHA under microscopic guidance on the back table. Patency was checked through the stump of the anterior branch of the RHA. With this technique, only one orifice, the stump of right anterior hepatic artery, was used for hepatic artery reconstruction. We have performed this technique in two patients. Both had good arterial flow after living donor liver transplantation. This innovative technique is easy and safe, and requires only one anastomosis, which, in theory, decreases the adds of developing hepatic arterial thrombosis.

Artificial vascular graft for inferior vena cava reconstruction in living donor liver transplantation: a case report.

Artificial grafts are not recommended because of the high incidence of thrombogenic effects. However, in some situations, such as emergency or when no vascular bank is available, an artificial graft must be used. We present a case in which a polytetrafluoroethyline graft was used as a conduit to reconstruct the retrohepatic vena cava severed during living donor liver transplantation (LDLT). A 48-year-old woman had end-stage primary biliary cirrhosis for 5 years received a right lobe liver graft from her son. The retrohepatic vena cava was divided and ligated in several sequences. The upper end of the severed retrohepatic vena cava retracted into the liver parenchyma. The lower end of the severed vena cava was distended, with multiple stitches. A 16-mm artificial graft was used as a conduit to replace the inferin vena cava for outflow reconstruction. The patient tolerated the complicated procedure well. No anticoagulant was used throughout the entire course. The patient has been well with excellent liver function after follow-up for more than 5 years. Magnetic resonance imaging and Doppler ultrasonographic studies showed good patency of the cava with no evidence of thrombosis. We suggest use of an artificial graft in living donor liver transplantation, in particular in urgent situations when autologous or allogeneic vessels are not available.

Polytetrafluoroethylene patch venoplasty for outflow reconstruction in living donor liver transplantation: two case reports.

Outflow obstruction may lead to liver congestion and eventual graft failure after living donor liver transplantation. Various methods of venoplasty provide wider outflow tracts. Most series have suggested use of autologous or allogenic grafts for patch venoplasty. We used a polytetrafluorethylene patch in two patients. Both showed good patency of the outflow tract at Doppler ultrasonography at 7 months and 4 months posttransplantation. A polytetrafluoroethylene patch may be a good alternative when no other autologous or allogeneic vascular patch is available or when the situation is critical.

Genetic algorithm for the efficient selection of disyllabic word lists used in Mandarin speech discrimination tests.

Speech audiometric tests have been widely used for advanced hearing diagnoses and in rehabilitation. However, there are no standardised speech tests for more than 90% of the world's population, who do not speak English. A major problem in the design of a speech audiometric test is that the selection of test materials is subject to multiple criteria, and its complexity rises dramatically as the structure of test items changes from phonemic or monosyllabic forms to disyllabic or polysyllabic forms. A genetic algorithm is presented that can automatically select a set of disyllabic words from a large Mandarin corpus. The selection accords with the following principal criteria for the items constituting a speech discrimination test: similarity in structure, familiarity to the subjects, and a phonemically balanced composition. The performance of the genetic algorithm was evaluated by computation of the distance between a target vector, specifying the desired distribution of initial and final syllables and tone patterns for daily disyllabic word usage, and the vector derived by the search results of the algorithm. The use of the genetic algorithm was illustrated by its application to the selection of test lists from two Mandarin corpora. The results showed that, for a given corpus, at least 12 disyllabic word lists with a distance of less than 20 could be generated within 72 h. The genetic algorithm performed an efficient, robust and low-complexity search of the problem space and can be easily modified to adapt to the material selection of other languages.