Intraoperative blood loss does not independently affect the survival outcome of gastric cancer patients who underwent curative resection.

BACKGROUND: Whether intraoperative blood loss (IBL) was independently associated with poor prognosis of gastric cancer (GC) patients remains controversial. In the present study, we evaluated the impact of IBL on the disease-free survival (DFS) of GC patients. METHODS: A total of 1669 patients who underwent curative gastrectomy for GC were reviewed retrospectively. All patients were classified as IBL < 400 mL and IBL ≥ 400 mL group according to the amount of IBL. The prognostic difference between two patient groups was compared and clinicopathologic factors associated with the prognosis of GC patients were analyzed. RESULTS: The 5-year DFS rate of the patients with IBL < 400 mL and those with IBL ≥ 400 mL was 52.1% and 41.5%, respectively (P < 0.001). The 5-year DFS rate of the patients who did and did not receive intraoperative blood transfusion was 36.9% and 53.2%, respectively (P < 0.001). However, the similar survival outcomes were not observed in the subgroup analysis based on the TNM stage. The multivariate analysis indicated that IBL (HR 1.021, 95% CI 0.875-1.191, P > 0.05) and intraoperative blood transfusion (HR 1.111, 95% CI 0.943-1.309, P > 0.05) were not independent prognostic factors for GC patients. In addition, the patients with IBL ≥ 400 mL had a higher risk of postoperative complications than those with IBL < 400 mL, especially for intraabdominal infection and wound infection. The tumor located in upper 1/3 stomach, total gastrectomy, combined organ resection and advanced tumor stage (stage III) were independent risk factors for intraoperative massive hemorrhage. CONCLUSION: Intraoperative blood loss was significantly associated with tumor-related and surgery-related factors. Intraoperative blood loss itself could not independently affect survival outcome of GC patients after curative gastrectomy.

Assessment of genetic relationship among Rhododendron cultivars using amplified fragment length polymorphism and inter-simple sequence repeat markers.

Genetic relationships of 17 Rhododendron cultivars, China, were assessed using inter-simple sequence repeat (ISSR) and amplified fragment length polymorphism (AFLP) markers. A total of 133 bands were obtained using nine selected ISSR primers, 129 (96.99%) of which were polymorphic; 267 bands were amplified by four AFLP primer pairs, 251 (94.01%) of which exhibited polymorphism. Based on these polymorphic products, a cluster analysis revealed similarities between the results of the ISSR and AFLP. All of the cultivars were clustered into two major branches; one branch contained the same four cultivars, and the other cultivars were separated into different groups in the other branch. The cluster results showed that the genetic relationships of the 17 cultivars were partly related to their morphological characteristics, particularly the flowering phase. Therefore, the results of this study support the classification of Rhododendron cultivars according to flowering phase. In addition, the cluster results can be used to select suitable parents for breeding.

Study of the expression of GABA(A) receptor in rats during acute lung injury caused by endotoxin.

The objective of the present study was to investigate the role of γ-aminobutyric acid type A receptor (GABA(A)R) in lipopolysaccharide (LPS)-induced acute lung injury (ALI) in rats. Thirty-two male wistar rats were randomly divided into four groups. Rats in the GABA group were pretreated with LPS and GABA, while those in the bicuculline (BIC) group were pretreated with LPS and bicuculline. We assessed the arterial blood gas, dry/wet ratio, and the level of tumor necrosis factor-α (TNF-α), IL-6, malondialdehyde, and superoxide dismutase 6 h after the immunization. Paraffin sections of samples were detected using the steptavidin-peroxidase method. Protein expression was detected using sodium dodecyl sulfate-polyacrylamide gel electrophoresis and western blotting. PaO2 in the LPS group was significantly lower than that in the control rats. Activation of GABA-mediated signaling by GABA increased the expression of GABA(A)R in airway bronchial and alveolar epithelial cells. Blockade of the GABA(A)R by bicuculline limited the expression of this receptor. The GABA group rats had higher levels of tissue TNF-α and IL-6 than in ALI rats and control rats. The BIC group rats demonstrated an opposite expression level compared to the GABA group rats. Our results suggest that the GABA(A)R could aggravate the inflammatory response syndrome and oxidative stress in the lungs and play an essential role in LPS-induced acute lung injury. It provides a novel method to study the incidence and mortality of ALI during the peroperative period.

Dopamine receptor D1 but not D3 essential for morphine-induced conditioned responses.

Recent studies indicate the involvement of dopamine receptors D1 and D3 in the regulation of locomotor stimulant and conditioned responses to morphine in mice. Moreover, expression of brain-derived neurotrophic factor (BDNF) may be modulated by D1 and D3 receptor activities in the nucleus accumbens (NAc) and prefrontal cortex (PFC). However, the underlying interactions between D1 and D3 receptors and BDNF in the expression of behavioral responses controlled by drug-associated cues have not yet been fully elucidated. In this study, we used dopamine receptor mutant mice to explore the roles of the D1 and D3 receptors in locomotion and morphine-induced place preference; furthermore, we investigated the effects of morphine on BDNF expression in the NAc and PFC of the mouse brain. Our results show that D1 receptor but not D3 receptor mutant mice had decreased sensitivity to acute morphine-induced (10 mg/kg) locomotion (D1: 3814.82 ± 319.9 cm vs D3: 8089.64 ± 967.4 cm). Furthermore, D1 receptor mutant mice did not acquire morphine-conditioned place preference (D1: -18.3 ± 59.9, D3: 217.7 ± 64.1) and showed decreased BDNF expression in the NAc (D1: 0.33 ± 0.07 fold, D3: 2.21 ± 0.18 fold) and PFC (D1: 0.74 ± 0.15 fold, D3: 1.68 ± 0.22 fold) compared with wild-type and D3 receptor mutant mice. These findings suggest that the D1 receptor is necessary for the induction of cue-associated morphine seeking and modulates locomotor habituation processes in response to acute morphine. The dopamine receptor D1 but not the D3 is also critical for morphine-induced BDNF expression in the NAc and PFC.

Advanced oxidation protein products as a biomarker of cutaneous lupus erythematosus complicated by nephritis: a case-control study.

Oxidative stress is involved in the pathogenesis of lupus nephritis (LN). The current study investigated the significance of advanced oxidation protein products (AOPPs) as a biomarker of LN in patients with cutaneous lupus erythematosus. Ninety-two patients who initially presented with systemic lupus erythematosus were divided into the LN- and LN+ groups. Serum AOPP levels were determined, and the association between AOPP levels and LN was investigated in a case-control study. In the LN+ group, patients with higher AOPP levels exhibited higher levels of dsDNA and proteinuria but lower levels of eGFR and complement C3 compared to those in patients with lower AOPP levels. A multivariable logistic regression model showed that the AOPP level was an independent risk factor for LN. The risk of nephritis specifically increased 24% for each 10 µM increase in AOPP (95% confidence interval, 1.166-1.915, P = 0.030). In contrast, neither elevated dsDNA level nor decreased complement C3 level was an independent risk factor for LN. Higher serum AOPP levels were associated with an increased risk of LN. Therefore, future studies are warranted to determine the potential clinical value of this novel biomarker.

Overregulation of microRNA-212 in the poor prognosis of esophageal cancer patients.

There have been few reports evaluating the expression and function of the microRNA miR-212 in esophageal cancer. The aim of this study was to investigate the relationship between miR-212 expression and clinicopathological factors and prognoses of esophageal cancer. MicroRNA was extracted from 46 esophageal cancer patients using the Taqman MicroRNA assay. All patients were at the same tumor node metastasis stage, but with different prognoses, and had all undergone surgery. The correlation between miR-212 expression and clinicopathological features was analyzed and the significance of miR-212 as a prognostic factor as well as its relationship with survival was determined. miR-212 expression was higher in patients with poor prognoses than in those with good prognoses (P < 0.0001). Kaplan-Meier analysis results showed that the miR-212 expression level was significantly correlated with survival time (P = 0.024). Patients with higher expression of miR-212 showed longer survival times. Cox multi-factor model analysis showed that miR-212 expression was significantly correlated with survival time (P = 0.026). mir-212 is related with prognostic factors and survival time and may be a biomarker for esophageal cancer.

Implications of microRNA-197 downregulated expression in esophageal cancer with poor prognosis.

The aim of this study was to investigate the significance of the microRNA miR-197 expression level in relation to clinicopathological factors and prognoses of esophageal cancer (EC). MicroRNA was extracted using the Taqman(®) MicroRNA Assay from 46 EC patients at the same tumor node metastasis (TNM) stage, but with different prognoses, who underwent surgery. Paracancerous normal tissues were used as controls. The correlation between miR-197 expression and clinicopathologic features was analyzed, and the significance of miR-197 as a prognostic factor and its relationship with survival was determined. miR-197 expression was lower in patients with poor prognosis than in those with good prognosis (P < 0.05). Kaplan-Meier analysis results showed that the miR-197 expression level is significantly correlated with survival time (P = 0.030), and that patients with higher expression of miR-197 had longer survival times. Cox multi-factor model analysis showed that patient prognosis (P = 0.001), tumor length (P = 0.010) and expression (P = 0.042), and survival time were significantly correlated, with corresponding risks of 9.183, 2.318, and 1.925, respectively. This study supports a role of miR-197 as an anti-oncogene and a biomarker for EC and its relationship with other prognostic factors and survival.

Association of a transforming growth factor-ß1 polymorphism with acute coronary syndrome in a Chinese Han population.

Acute coronary syndrome (ACS) is a complex multifactorial and polygenic disorder that is thought to result from the interaction between an individual's genetic makeup and various environmental factors. The aim of this study was to investigate the association of a transforming growth factor-ß1 (TGF-ß1) polymorphism (-509C>T) with ACS in a Chinese Han population. The TGF-ß1 polymorphism was evaluated in 336 patients with ACS and 396 healthy control subjects by polymerase chain reaction-restriction fragment length polymorphism. The genotype distributions of the control and ACS groups were in Hardy-Weinberg equilibrium (X(2) = 3.54 and X(2) = 1.72, respectively, P > 0.05). The frequencies of the CC, CT, and TT genotypes were 22.61, 53.57, and 20.83% in the ACS group, respectively, whereas they were 8.33, 48.74, and 42.17% in controls. There were significant differences between controls and ACS patients in the frequencies of the CC genotype and the C allele. These results suggest that the promoter polymorphism (-509C>T) in TGF-ß1 is associated with ACS in this population. The CC genotype and the C allele of TGF-ß1 might be a specific risk factor of ACS in the Chinese Han population in Xinjiang.

Molecular cloning and expression pattern of the DjStag gene in the planarian Dugesia japonica during embryonic development.

We examined STAG-related gene (DjStag) expression in the planarian Dugesia japonica. This species is common in Far Eastern countries. The DjStag cDNA includes 1362 bp and contains a 489-bp open reading frame corresponding to a deduced protein of 162 amino acids, with a 170-bp 5'-UTR and a 703-bp 3'-UTR. Phylogenetic analysis showed that DjStag is an STAG/STAG-like member. We examined the expression pattern of DjStag in this planarian during embryonic development by whole-mount in situ hybridization. DjStag was detected in embryonic cells in the germ band at early embryo stages. The number of DjStag-positive embryonic cells increased in stage 5. Later, it was mainly expressed in lateral region parenchyma. In juveniles, extensive expression of DjStag was observed not only in the head and tail regions, but also in the parenchyma between the epidermis and the gastrodermis. We conclude that DjStag is expressed in the cellular subset that will become the neoblast cells of the adult flatworm. DjStag may play an essential role in spatial and temporal regulation during planarian embryonic development.

SU-E-T-397: Interplay Effect of Gated Lung Stereotactic Body Radiotherapy with RapidArc Delivery.

PURPOSE: To investigate the dosimetric effect of intrafraction tumor motion during gated RapidArc Stereotactic Body Radiotherapy (SBRT) delivery. METHOD: The realtime tumor motion data were retrieved from 6 lung patients. Each of them received 3 fractions of stereotactic radiotherapy treatments with Cyberknife Synchrony. Phase gating through an external surrogate was simulated with a gating window of 5 mm. The resulting residual tumor motion curves during gating (beam-on) were retrieved. RapidArc SBRT was planned on the platform of Varian Truebeam at 6 MV with 1400 MU/min. Planning target volume (PTV) was defined as physician-contoured clinical target volume (CTV) surrounded by an isotropic 5 mm margin. Each patient was prescribed with 60Gy/3 fractions. The RA plan typically consists of 2 arcs; each contains 90-120 control points. An algorithm was developed to reconstruct the delivered dose with tumor motion. The MLC segment is assumed to move relatively to a static tumor. Each MLC control point, mainly the leaf position were modified according to the probability density function of tumor motion. The newly created MLC control points were written back to the treatment file in the dicom format which was subsequently imported to treatment planning system (Varian Eclipse) for dose recalculation. RESULTS: The magnitude of dose deviation with motion is consistent with the excursion of the residual tumor movement. Overall CTV coverage of the study group is barely affected owing to the 5 mm margin. The fractional PTV dose coverage dropped by 4% at most and that from all fractions by 3%. An examination in the point dose shows an increase of 4% in the maximum dose and decrease of 10% for the minimum dose. CONCLUSION: With effective gating, interplay effect does not change the target coverage much during gated RapidArc SBRT. However it increases the dose nonuniformity inside target.

SU-E-T-636: Comparison of RapidArc-Based Radiosurgery with Cone-Based Cyberknife Treatment for Multiple Intracranial Tumors.

PURPOSE: To investigate the feasibility of RapidArc technique on intracranial radiosurgery for multiple lesions. METHODS: Six patients who were previously treated using cone-based technique, Cyberknife were included in this study. These patients have multiple lesions (6-9, mostly metastasis). In our current clinical practice, each lesion was planned and treated individually. The prescription was 15-21 Gy at 80% with single fraction. These cases were replanned with RapidArc on the platform of Varian Truebeam STx equipped with high resolution MLC leaves of 2.5mm at center. The maximum dose rate is 1400 MU/min at 6 MV for flattering filter free mode. Because of long span of multiple lesions, the targets were divided into two groups with two isocenters. Each plan with one isocenter contains 4 non-coplanar arcs, and dose optimization was performed with the two plans combined. Critical organs, such as eyes, brainstem and brain were constrained. The individual Cyberknife plans were summed to compare with the RapidArc plan. Scenarios of setup error were simulated during RapidArc treatment. RESULTS: RapidArc plans can achieve comparable target coverage and normal tissue avoidance to Cyberknife plans. The brain dose volume histogram (DVH) curves of the two techniques are similar in spite of different appearance of their 3D dose distributions. MU is much higher for summed Cyberknife plan. Because RapidArc can treat several lesions together, the complete treatment time for all lesions is significantly reduced. However RapidArc treatment is susceptible to setup error, which may cause increase in normal tissue dose and decrease in target dose coverage. The level of discrepancy depends on the magnitude of setup error, location and dose distribution of the target. CONCLUSION: Multiple brain lesions treatment with RapidArc radiosurgery is clinically feasible with setup error fully accounted. It can provide dose performance comparable to cone-based Cyberknife treatment.

Expression patterns of the STAG gene in intact and regenerating planarians (Dugesia japonica).

We examined the spatial and temporal expression of the planarian Dugesia japonica STAG-related gene (DjStag), in both intact and regenerating planarians, by whole-mount in situ hybridization and relative quantitative real-time PCR. The first localized transcripts of DjStag were detected in the blastemas three days after amputation, in all regenerates including those from head, tail and trunk pieces. The maximum level of expression of DjStag transcripts occurred at five days after cutting. After regeneration for seven days, DjStag was weakly expressed. A similar decrease occurs regardless of the orientation of the cut. The expression pattern did not differ significantly in the different types of regeneration. Relative quantitative real-time PCR analysis of DjStag mRNA indicated that the expression of DjStag mRNA was increased after amputation compared to that in normal intact planarians, and the maximum level of expression of DjStag transcripts occurred at five days after amputation. All results suggest that DjStag, implicated in planarian regeneration, plays a role in maintaining the ability of pluripotent stem cells to regenerate lost tissue in planarians.

Spectroscopic studies on the interaction of efonidipine with bovine serum albumin.

Efonidipine hydrochloride is an antihypertensive and antianginal agent with fewer side effects and is better tolerated in the treatment of hypertension with renal impairment. Its interaction with bovine serum albumin (BSA) is of great use for the understanding of the pharmacokinetic and pharmacodynamic mechanisms of the drug. The binding of efonidipine to BSA was investigated by fluorescence spectroscopy and circular dichroism. BSA fluorescence was quenched by efonidipine, due to the fact that efonidipine quenched the fluorescence of tryptophan residues mainly by the collision mode. The thermodynamic parameters DeltaH0 and DeltaS0 were 68.04 kJ/mol and 319.42 J x mol-1 x K-1, respectively, indicating that the hydrophobic interactions played a major role. The results of circular dichroism and synchronous fluorescence measurements showed that the binding of efonidipine to BSA led to a conformational change of BSA. The fraction of occupied sites (theta) for the 8-anilino-1-naphthalein-sulfonic acid (ANS)-BSA system is 85%, whereas for the NZ-105-BSA system, it is 53%, which suggests that the interaction of ANS with BSA is stronger than that of NZ-105 with BSA. Binding studies in the presence of ANS indicated that efonidipine competed with ANS for hydrophobic sites of BSA. The effects of metal ions on the binding constant of the efonidipine-BSA complex were also investigated. The presence of metal ions Zn2+, Mg2+, Al3+, K+, and Ca2+ increased the binding constant of efonidipine-BSA complex, which may prolong the storage period of NZ-105 in blood plasma and enhance its maximum effects.

Spectroscopic studies on the interaction of efonidipine with bovine serum albumin

Efonidipine hydrochloride is an antihypertensive and antianginal agent with fewer side effects and is better tolerated in the treatment of hypertension with renal impairment. Its interaction with bovine serum albumin (BSA) is of great use for the understanding of the pharmacokinetic and pharmacodynamic mechanisms of the drug. The binding of efonidipine to BSA was investigated by fluorescence spectroscopy and circular dichroism. BSA fluorescence was quenched by efonidipine, due to the fact that efonidipine quenched the fluorescence of tryptophan residues mainly by the collision mode. The thermodynamic parameters ÄH0 and ÄS0 were 68.04 kJ/mol and 319.42 J·mol-1·K-1, respectively, indicating that the hydrophobic interactions played a major role. The results of circular dichroism and synchronous fluorescence measurements showed that the binding of efonidipine to BSA led to a conformational change of BSA. The fraction of occupied sites (è) for the 8-anilino-1-naphthalein-sulfonic acid (ANS)-BSA system is 85 percent, whereas for the NZ-105-BSA system, it is 53 percent, which suggests that the interaction of ANS with BSA is stronger than that of NZ-105 with BSA. Binding studies in the presence of ANS indicated that efonidipine competed with ANS for hydrophobic sites of BSA. The effects of metal ions on the binding constant of the efonidipine-BSA complex were also investigated. The presence of metal ions Zn2+, Mg2+, Al3+, K+, and Ca2+ increased the binding constant of efonidipine_BSA complex, which may prolong the storage period of NZ-105 in blood plasma and enhance its maximum effects.