Multiple variants in 5q31.1 are associated with systemic lupus erythematosus susceptibility and subphenotypes in the Han Chinese population.

BACKGROUND: A previous study provided evidence for a genetic association between PPP2CA on 5q31.1 and systemic lupus erythematosus (SLE) across multi-ancestral cohorts, but failed to find significant evidence for an association in the Han Chinese population. OBJECTIVES: To explore the association between this locus and SLE using data from our previously published genome-wide association study (GWAS). METHODS: Single-nucleotide polymorphisms (SNPs) rs7726414 and rs244689 (near TCF7 and PPP2CA in 5q31.1) were selected as candidate independent associations from a large-scale study in a Han Chinese population consisting of 1047 cases and 1205 controls. Subsequently, 3509 cases and 8246 controls were genotyped in two further replication studies. We then investigated the SNPs' associations with SLE subphenotypes and gene expression in peripheral blood mononuclear cells. RESULTS: Highly significant associations with SLE in the Han Chinese population were detected for SNPs rs7726414 and rs244689 by combining the genotype data from our previous GWAS and two independent replication cohorts. Further conditional analyses indicated that these two SNPs contribute to disease susceptibility independently. A significant association with SLE, age at diagnosis < 20 years, was found for rs7726414 (P = 0·001). The expression levels of TCF7 and PPP2CA messenger RNA in patients with SLE were significantly decreased compared with those in healthy controls. CONCLUSIONS: This study found evidence for multiple associations with SLE in 5q31.1 at genome-wide levels of significance for the first time in a Han Chinese population, in a combined genotype dataset. These findings suggest that variants in the 5q31.1 locus not only provide novel insights into the genetic architecture of SLE, but also contribute to the complex subphenotypes of SLE.

[Role of neutrophils in treatment of rats with D-galactosamine-induced acute liver failure with bone marrow mesenchymal stem cells].

Objective: To investigate the therapeutic effect of bone marrow mesenchymal stromal cell (BMSC) transplantation on D-galactosamine-induced acute liver failure in Sprague-Dawley (SD) rats, as well as the mechanism of neutrophils in this process. Methods: A total of 39 male SD rats were divided into control group (8 rats, intraperitoneal injection of isotonic saline), model group (10 rats, intraperitoneal injection of D-galactosamine), solvent group (9 rats, tail vein injection of isotonic saline at 2 hours after intraperitoneal injection of D-galactosamine), and treatment group (12 rats, tail vein injection of MSCs at 2 hours after intraperitoneal injection of D-galactosamine). The rats were sacrificed at 24 hours after the model of D-galactosamine-induced acute liver failure was established, and the blood and liver tissue were harvested. The serum levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) and total bilirubin (TBil) were measured, and blood analysis was performed to measure the number and percentage of neutrophils in peripheral blood. Immunofluorescence assay was used to measure the expression of the neutrophil marker Ly6g in the liver, the myeloperoxidase (MPO) kit was used to measure the activity of MPO in liver, and RT-PCR was used to measure the mRNA expression of inflammatory cytokines and chemokines in the liver, i.e., tumor necrosis factor-α(TNF-α), interleukin-1ß(IL-1ß), interferon-γ(IFN-γ), interleukin-10 (IL-10), CXC chemokine ligand 1 (CXCL1), and CXC chemokine ligand 2 (CXCL2). Another 64 male SD rats were randomly divided into groups, and the survival rates of rats in each group were observed for 7 days. The independent samples t-test was used for comparison between any two groups (Levene homogeneity test of variance, and the corrected t-test was used for a P value of < 0.05), and the log-rank test was used for comparison of survival rates between any two groups. Results: At 24 hours after acute liver failure was induced by D-galactosamine in the SD rats, there were significant increases in the liver function parameters (ALT: 2884.1±541.0 U/L vs 45.4±11.0 U/L,P< 0.001; AST: 3634.9±755.9 U/L vs 143.9±23.7 U/L,P< 0.001; TBil: 44.4±8.4µmmol/L vs 0.9±0.2µmmol/L,P< 0.001) and the number and percentage of peripheral blood neutrophils [number: (4.7±1.1)×109 vs (1.4±0.4)×109,P< 0.001; percentage: 44.9%±8.0% vs 18.3%±4.4%,P< 0.001]. A large number of neutrophils aggregated in the liver tissue, and there were significant increases in the MPO activity (4.72±1.09 U/g vs 1.13±0.24 U/g,P< 0.001), inflammatory cytokines, and chemokines. Compared with the model group, the treatment group showed significant improvements in liver function (ALT: 1 823.9±389.2 U/L vs 2 884.1±541.0 U/L,P< 0.001; AST: 2173.0±567.3 U/L vs 3634.9±755.9 U/L,P< 0.001; TBil: 30.9±6.5µmmol/L vs 44.4±8.4µmmol/L,P< 0.001) and survival rate (50% vs 12.5%,P= 0.023). Meanwhile, the treatment group also showed significant reductions in the number and percentage of peripheral blood neutrophils [number: (3.5±1.0)×109 vs (4.7±1.1)×109,P= 0.012; percentage: 35.9%±8.9% vs 44.9%±8.0%,P= 0.021], number of neutrophils in the liver, and MPO activity (3.52±1.03 U/g vs 4.72±1.09 U/g,P= 0.040), as well as significantly inhibited expression of inflammatory cytokines and chemokines (TNF-α: 2.458±0.762 vs 3.778±1.046, P = 0.005; IL-1ß: 2.498±0.547 vs 4.065 ± 0.953,P= 0.002; IFN-γ: 3.977±1.039 vs 5.418±1.255, P = 0.025; IL-10: 6.056±1.542 vs 3.368±0.952,P= 0.001; CXCL1: 7.988±1.911 vs 10.366±1.239,P= 0.010; CXCL2: 3.441±1.005 vs 4.847±1.113,P= 0.019). Conclusion: BMSC transplantation has a therapeutic effect on D-galactosamine-induced acute liver failure in rats, and this process is accompanied by reduced aggregation and activity of neutrophils in peripheral blood and liver. Inflammatory cytokines and chemokines may be involved in the mechanism of regulation of these two aspects.

Systematic review and meta-analysis of triclosan-coated sutures for the prevention of surgical-site infection.

BACKGROUND: Surgical-site infections (SSIs) increase morbidity and mortality in surgical patients and represent an economic burden to healthcare systems. Experiments have shown that triclosan-coated sutures (TCS) are beneficial in the prevention of SSI, although the results from individual randomized controlled trials (RCTs) are inconclusive. A meta-analysis of available RCTs was performed to evaluate the efficacy of TCS in the prevention of SSI. METHODS: A systematic search of PubMed, Embase, MEDLINE, Web of Science(®), the Cochrane Central Register of Controlled Trials and internet-based trial registries for RCTs comparing the effect of TCS and conventional uncoated sutures on SSIs was conducted until June 2012. The primary outcome investigated was the incidence of SSI. Pooled relative risks with 95 per cent confidence interval (c.i.) were estimated with RevMan 5.1.6. RESULTS: Seventeen RCTs involving 3720 participants were included. No heterogeneity of statistical significance across studies was observed. TCS showed a significant advantage in reducing the rate of SSI by 30 per cent (relative risk 0·70, 95 per cent c.i. 0·57 to 0·85; P < 0·001). Subgroup analyses revealed consistent results in favour of TCS in adult patients, abdominal procedures, and clean or clean-contaminated surgical wounds. CONCLUSION: TCS demonstrated a significant beneficial effect in the prevention of SSI after surgery.

Posterior second sacral alar iliac screw insertion: anatomic study in a Chinese population.

OBJECTIVE: To provide radiographic parameters for optimal placement of posterior second sacral alar iliac (S2AI) screw for instrumentation and fusion of scoliosis to the second sacral level in a Chinese population. METHODS: S2AI screw trajectories were mapped on three-dimensional computed tomography (3DCT) reconstructions of 60 normal adult pelvises. 1 mm inferior and 1 mm lateral to the S1 dorsal foramen were chosen as the entry point, and ideal S2AI screw trajectories were explored by rotating and cutting the 3D pelvis, ensuring that the trajectories were of maximum length and width. The directions and depth of these determined trajectories were then measured. RESULTS: The ideal S2AI screw trajectories could be found in each pelvis. The left and right screw trajectory parameters for males were shown as follows: angulation was L 29.15 ± 8.60° vs. R 29.96 ± 8.28° (p = 0.286) caudally in the sagittal plane and L 36.49 ± 3.14° vs. R 37.16 ± 3.14° (p = 0.165) laterally in the transverse plane. The maximal and intrasacral lengths of trajectory were L 121.25 ± 8.33 vs. R 120.63 ± 7.54 mm (p = 0.460) and L 26.20 ± 3.31 vs. R 26.92 ± 4.76 mm (p = 0.268). The entry point was L 28.87 ± 3.33 vs. R 29.79 ± 3.55 mm (p = 0.186) lateral to the second sacral midline, and L 44.14 ± 11.87 vs. R 43.89 ± 12.53 mm (p = 0.687) underneath the skin. The trajectories for females were more caudal (L: 34.50 ± 6.56° vs. 29.15 ± 8.60°, p = 0.009; R: 35.72 ± 7.53° vs. 29.96 ± 8.28°, p = 0.007) in the sagittal plane, but the lateral angulation in the transverse plane showed no difference between genders (p > 0.05). The female iliac medullar cavities were obviously narrower than those of males (L: 14.76 ± 2.46 vs. 16.98 ± 3.52, p = 0.006; R: 14.94 ± 2.60 vs. 17.00 ± 2.81, p = 0.005). Although the average maximal length of trajectories for females were about 5 mm shorter than those of males, intrasacral length were equal to those of males. Furthermore, both the distance from entry point to the S2 midline and skin in the transverse plane showed no difference between genders. CONCLUSION: The feasibility to insert S2AI screws to the sacrum and ilium in an Asian population along with the ideal entry angle and length of trajectory were identified for clinical practice.

RNA-electroporated CD40-activated B cells induce functional T-cell responses against HepG2 cells.

Hepatocellular carcinoma (HCC) is one of the incurable tumours in the world. Cell-based immunotherapy, in which antigen-loaded antigen-presenting cells (APCs) are able to elicit T cell responses, has become an alternative treatment for liver cancer. Here, we used HepG2 cells' total RNA-electroporated CD40 ligand-activated B (CD40-B) cells as alternative APC for induction of specific CD8+ T-cell responses. The antigen-presenting ability of CD40-B cells was determined by phenotypic analysis, showing a polyclonal, strongly activated B-cell population with high expression of co-stimulatory molecules. To demonstrate the ability of total RNA extracted from HepG2 cells electroporated CD40-B cells to induce CD8+ T-cell responses, these RNA-loaded cells were co-cultured with autologous peripheral blood mononuclear cells for 7 days followed by analysis of T-cell antigen specificity. These experiments showed that CD40-B cells electroporated with HepG2 cells' total RNA are capable of activating antigen-specific interferon-gamma-producing CD8+ T cells, and these T cells activated by CD40-B cells show a killing effect on HepG2 cells. These findings demonstrated that the carcinoma cell derived total RNA-electroporated CD40-B cells could be used as alternative APC for the induction of antigen-specific CD8+ T-cell responses, which might be used in HCC immunotherapy.

Liver progenitor cells activated after 30% small-for-size liver transplantation in rats: a preliminary study.

AIM: To explore whether liver progenitor cells were activated after 30% small-for-size liver transplantation in rats. METHODS: 200 rats were arranged in three groups: 70% partial hepatectomy (PH), whole liver transplantation (WLT), and 30% liver transplantation (SLT) group. On days 1, 2, 3, 5, 7 after operation, 6 rats were sacrificed in each group at each time. One week survivals were analyzed; while liver injury and regeneration index were estimated by serum ALT AST, H&E staining and proliferating cell nuclear antigen index. The oval cell markers, including CD90 and OV6, were detected in liver sections by immunohistochemistry. RESULTS: The 50% survival rate of the SLT group was significantly lower than those of the PH and the WLT groups. At each time after operation, the serum ALT and AST were much higher in the SLT group. Compared with the PH group on days 1, 2, and 3 postoperatively, the PCNA indices were lower among the SLT group. OV-6 positive and CD90 positive cells were detected in the SLT group from day 2 postoperatively. These progenitor cells were first dispersed in the liver but restricted to the periportal region over the following days. CONCLUSION: Liver progenitor cell activation after SLT may be related to the liver dysfunction caused by a small-for-size graft.

A novel model for external biliary drainage in rat orthotopic liver transplantation.

OBJECTIVE: The objective of this study was to establish a stable model of orthotopic liver transplantation with external biliary drainage in rats, to provide the possibility for dynamic detection of bile characteristics after transplantation. METHODS: Ninety-eight Sprague Dawley rats were divided randomly into a control group (n = 30) and an external drainage group (n = 68). Biliary extra-drainage group rats were established as follows: an epidural catheter was implanted into the common bile duct and an appropriate hose was connected from the outside. A stoma was made into the high jejunum, and a hose was implanted. Then the 2 hoses were induced to connect with each other in the subcutis of the right lateral abdominal wall. RESULTS: The operative success rate and 1-week and 2-week survival rates of the extra-drainage group were 94.1% (64/68), 88.2% (60/68), and 79.4% (54/68), respectively. Meanwhile, in the control group, the operative success rate and 1-week and 2-week survival rates were 93.3% (28/30), 90% (27/30), and 83.3% (25/30), respectively. There were no significant differences at any time between the 2 groups (p > 0.05). CONCLUSIONS: The biliary extra-drainage model in rat orthotopic liver transplantation provides a novel, simple, and reliable method for dynamic collection of bile. The operation success rate was satisfactory. This model was successful and could be applied in various experimental studies.

[Normothermic total hepatic vascular exclusion for hepatectomy].

Experience of normothermic total hepatic vascular exclusion for hepatectomy in two patients with hepatocarcinoma in the second porta hepatis area are reported. The combined method of stepwise exclusion of porta hepatis, total hepatic vascular exclusion, total hepatic vascular exclusion without abdominal aorta and manipulative exclusion of inferior vena cava above liver are described. Biochemical studies before, during and after the operation showed that they are similar to these of routine hepatectomy. This modification, being safer and easier, may increase the resectability and reduce the mortality rate.

Effects of orthotopic liver transplantation in inbred rats on bone biomechanical properties.

AIM: To determine the effects of orthotopic liver transplantation in inbred rats on the mechanical properties of bones at different anatomic sites. METHODS: The 24 rats that survived liver transplantation were paired with sham-operated rats of similar body weight. Six months after surgery, the lumbar vertebra, the proximal femur, and the middle femoral shaft were measured for their biomechanical properties and bone mineral density. RESULTS: The ultimate force, the ultimate stress, the Young modulus and the bone mineral density of both the proximal femur and the middle femur shaft of the rats were significantly reduced in the liver transplant group. However, no significant change was observed in the various parameters that indicate the biomechanical properties and the bone mineral density of the lumbar vertebra. CONCLUSIONS: Orthotopic liver transplantation impairs the biomechanical properties of the proximal femur and the middle femoral shaft. Orthotopic liver transplantation itself is one of the risk factors for posttransplant fracture.