To study the effect of nicorandil pretreatment on ketone body metabolism and Acetyl-CoA acetyltransferase (ACAT1) activity in hypoxia/reoxygenation (H/R)-induced cardiomyocytes. In our study, we applied H9c2 cardiomyocytes cell line to evaluate the cardioprotective effects of nicorandil. We detected mitochondrial viability, cellular apoptosis, reactive oxygen species (ROS) production and calcium overloading in H9c2 cells that exposed to H/R-induced cytotoxicity. Then we evaluated whether nicorandil possibly regulated ketone body, mainly β-hydroxybutyrate (BHB) and acetoacetate (ACAC), metabolism by regulating ACAT1 and Succinyl-CoA:3-keto-acid coenzyme A transferase 1 (OXCT1) protein and gene expressions. Nicorandil protected H9c2 cardiomyocytes against H/R-induced cytotoxicity dose-dependently by mitochondria-mediated anti-apoptosis pathway. Nicorandil significantly decreased cellular apoptotic rate and enhanced the ratio of Bcl-2/Bax expressions. Further, nicorandil decreased the production of ROS and alleviated calcium overloading in H/R-induced H9c2 cells. In crucial, nicorandil upregulated ACAT1 and OXCT1 protein expressions and either of their gene expressions, contributing to increased production of cellular BHB and ACAC. Nicorandil alleviated cardiomyocytes H/R-induced cytotoxicity through upregulating ACAT1/OXCT1 activity and ketone body metabolism, which might be a potential mechanism for emerging study of nicorandil and other K(ATP) channel openers.
Acetyl-CoA C-Acetyltransferase ; Apoptosis ; Calcium ; Cell Line ; Coenzyme A ; Gene Expression ; Metabolism* ; Myocytes, Cardiac ; Nicorandil* ; Reactive Oxygen Species ; Transferases

Objective To analyze the displacement of titanium clips for tumor bed localization after breast-conserving surgery for breast cancer and its influential factors.Methods A retrospective analysis was performed on the cone-beam computed tomography (CT) images of 14 patients with breast cancer who received radiotherapy after breast-conserving surgery from April to October,2016.The relative position of the chest wall and the errors of the titanium clips in radiotherapy were measured.A Pearson correlation analysis was used to analyze the correlation of the displacement of titanium clips with the relative position of titanium clips,the breast volume,the vertical distance between the titanium clips and the tangential line of the chest wall,and the maximum thickness of the breast.Results The system errors of the chest wall in left-right,superior-inferior,and anterior-posterior directions were 4.42,3.44,and 5.13 mm,respectively,and the random errors were 3.55,3.07,and 4.54 mm,respectively.The titanium clips had a large displacement relative to the chest wall,mainly in the left-right direction.The maximum system error was 4.39 mm and the random error was 2.42 mm.The displacement of titanium clips was not significantly correlated with the breast volume and the maximum thickness of the breast (P>0.05).However,the relative position of titanium clips in superior-inferior direction was significantly correlated with the displacement of the lowest,the most lateral,the most anterior,and the most posterior titanium clips (P<0.05).As to the uppermost clips,there was a significant difference in displacement between the clips close to the chest wall and the clips far from the chest wall (P=0.02).Conclusions Due to large setup error and displacement of titanium clips during radiotherapy,simultaneous integrated boost is not suitable for patients with breast cancer who are immobilized by vacuum cushion and received radiotherapy.The unstable immobilization may be the major influential factor for the displacement of titanium clips.

The affiliation of the second author, Lei Sen Han, should be corrected.

OBJECTIVETo understand the clinical features and prognosis of IgA nephropathy (IgAN) patients with glomerular crescents.

METHODSThe clinical data collected at the time of renal biopsy and the follow-up data of 89 IgAN patients with glomerular crescents were analyzed with 412 IgAN patients without crescents serving as the control group. Follow-up study was conducted in 78 patients with crescents and 198 without it, and the renal survival rate was estimated using Kaplan-Meier survival analysis.

RESULTSThe incidence of glomerular crescents was 17.8% in IgAN patients. Clinically, 39 patients with crescents experienced rapidly progressive glomerulonephritis, resulting in a significantly higher rate of this manifestation than that in patients without crescent. Patients with crescents also had more grave pathological changes in the glomerulus and renal tubule interstitium than the control patients. Patients were followed up for an average of 40.3-/+29.6 months in crescent group and 45.1-/+26.9 months in the control group, and the 1-, 3-, 5-year renal survival rate was 95.24%, 80.95%, 61.9% in the former and 100%, 91.67%, 79.17% in the latter, respectively.

CONCLUSIONIgAN patients with crescents have severer clinical and pathological manifestations and poorer prognosis than those without crescents.

Adolescent ; Adult ; Aged ; Child ; China ; epidemiology ; Female ; Follow-Up Studies ; Glomerulonephritis, IGA ; mortality ; pathology ; Humans ; Kaplan-Meier Estimate ; Kidney Glomerulus ; pathology ; Male ; Middle Aged ; Prognosis ; Survival Rate

A prokaryotic expression plasmid containing VIP (vasoactive intestinal peptide) and sTNFRII(soluble tumor necrosis factor receptor II ) genes was constructed. The sTNFRII was cloned by PCR by using special primers which contained VIP gene ORF and a linker in its forward primer. The amplified fragment was inserted into the expression vector pET32a between BamHI and Hind III restriction sites. Transformed E.coli DH5 by pET32a-VIP- sTNFRIIexpressed the fusion protein. After being identified, the protein was purified by ion exchange chromatography and by hydrophobic interaction chromatography. The reconstructed protein showed high bio-activity and could be applied for further use.

【Objective】 To investigate the association between fetal fraction(FF) of cell-free DNA and adverse pregnancy outcomes. 【Methods】 A retrospective case-control study was conducted in 1 231 Chinese pregnant women who underwent non-invasive prenatal testing and gave birth to their children in the Third Affiliated Hospital of Sun Yatsen University during January 2017 to October 2018, including HDP group(n = 84), FGR group(n = 57) and preterm birth group(n = 59) as case groups, and the pregnant women without pregnancy-related complications were included in the control group(n = 1 031). The correlation between FF and maternal age, gestational weeks, body mass index(BMI), and comparison of FF difference in different groups were analyzed. 【Results】 In 1 031 normal pregnant women performed NIPT, the plasma FF was(12.03 ± 3.64) %, FF and maternal BMI were negatively correlated(rs = -0.079, P < 0.05). FF in HDP, FGR, preterm birth group were(11.21 ± 2.60) %, (12.48 ± 3.92) %, (11.66 ± 3.34) % , respectively. Compared with control group, FF in HDP group was statistically lower, OR = 0.926, 95%CI: 0.859-0.997, P = 0.043. FF in FGR group and preterm birth group were not statistically different compared with control group. 【Conclusions】 Low FF is a risk factor of HDP, and we may apply FF of cell-free DNA to predict the risk of HDP in further research. The correlation of FF and FGR, preterm birth have not been found yet.

OBJECTIVETo determine the prevalence of beta-fibrinogen gene -455G/A, -148C/T polymorphisms in Chinese Han population and to investigate whether they were associated with pulmonary thromboembolism (PTE).

METHODSThe subjects consisted of 101 patients with PTE and 101 healthy controls matched with age and sex, from the same geographic area. All patients were diagnosed by high probability of lung ventilation/perfusion scan and/or multi-slice CT pulmonary angiography as well as medical history and clinical manifestations. Genome DNA was extracted from whole blood using KI-phenol-chloroform. Genotypes and allele frequencies of fibrinogen beta gene -455G/A, -148C/T polymorphisms were examined by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). Restriction enzyme HaeIII and HindIII digestion were used for detecting -455G/A, -148C/T polymorphisms respectively.

RESULTSRegarding fibrinogen beta gene -455G/A and -148C/, the allele frequencies G and A of fibrinogen beta -455 in the controls were 0.931, 0.069 while C and T of -148 were 0.777, 0.223 respectively, which were in good agreement with Hardy-Weinberg equilibrium. There was significant difference of -455G/A genotype frequencies distribution of AA, GA, GG between cases and in controls respectively, but no significant difference was found in the -148C/T polymorphisms. The frequencies of mutation allele -455A were 0.193, 0.169 in cases and in controls with P < 0.05 but there was no statistically significant difference of -148T allele. The presence of A allele of fibrinogen beta -455 was found to be a greater risk factor in cases than in controls. The odds ratio (OR) of GA and GA + AA were 3.723 (1.786 - 7.759), 3.749 (1.842 - 7.630), respectively. When compared with GG genotype, the P value was 0.0001.

CONCLUSIONThere was a complete linkage disequilibrium between fibrinogen beta -148C/T and -455G/A found. The frequencies of -455A, alleles in PTE disease were apparently higher than that of healthy adults but there was no difference in -148T alleles.

Asian Continental Ancestry Group ; genetics ; Case-Control Studies ; China ; Fibrinogen ; genetics ; Gene Frequency ; Genetic Predisposition to Disease ; Genotype ; Humans ; Linkage Disequilibrium ; Odds Ratio ; Polymerase Chain Reaction ; Polymorphism, Genetic ; Pulmonary Embolism ; genetics ; Risk Factors

BACKGROUNDThe cell layer of the ciliary epithelium is responsible for aqueous humor secretion and maintenance. Ion channels play an important role in these processes. The main aim of this study was to determine whether the well-characterized members of the Kv1 family (Kv1.3) contribute to the Kv currents in ciliary epithelium.

METHODSNew Zealand White rabbits were maintained in a 12 hours light/dark cycle. Ciliary epithelium samples were isolated from the rabbits. We used Western blotting and immunocytochemistry to identify the expression and location of a voltage-gated potassium channel Kv1.3 in ciliary body epithelium. Membrane potential change after adding of Kv1.3 inhibitor margatoxin (MgTX) was observed with a fluorescence method.

RESULTSWestern blotting and immunocytochemical studies showed that the Kv1.3 protein expressed in pigment ciliary epithelium and nonpigment ciliary epithelium, however it seemed to express more in the apical membrane of the nonpigmented epithelial cells. One nmol/L margatoxin, a specific inhibitor of Kv1.3 channels caused depolarization of the cultured nonpigmented epithelium (NPE) membrane potential. The cytosolic calcium increased after NPE cell depolarization, this increase of cytosolic calcium was partially blocked by 12.5 micromol/L dantrolene and 10 micromol/L nifedipine. These observations suggest that Kv1.3 channels modulate ciliary epithelium potential and effect calcium dependent mechanisms.

CONCLUSIONKv1.3 channels contribute to K+ efflux at the membrane of rabbit ciliary epithelium.

Animals ; Blotting, Western ; Calcium ; metabolism ; Ciliary Body ; cytology ; metabolism ; physiology ; Immunohistochemistry ; In Vitro Techniques ; Kv1.3 Potassium Channel ; metabolism ; physiology ; Membrane Potentials ; physiology ; Pigment Epithelium of Eye ; cytology ; metabolism ; physiology ; Rabbits

OBJECTIVETo study the relation of tumor interstitial T lymphocyte subset activity to the clinical staging of non-small-cell lung cancer (NSCLC) and the immune response.

METHODSImmunohistochemical staining for CD4(+), CD8(+) and CD4(+)CD25(+) Foxp3(+) (regulatory T cells, Treg) T cells was performed on paraffin-embedded tissues from 60 NSCLC cases.

RESULTSCompared to stage I/II NSCLC patients, patients in stage III/IV showed a significant decrease in the percentage of CD4(+) and CD4(+)/CD8(+) T cells (P<0.05) and an increase in CD8(+) and CD4(+)CD25(+) Foxp3(+) T cells (P<0.05). Treg cells were enriched in the tumor tissue as compared with those in the adjacent tissues.

CONCLUSIONSThe proportion of CD4(+)CD25(+) Foxp3(+) Treg cells is positively correlated to the clinical staging of NSCLC, in which T cell-mediated immune response is suppressed.

Aged ; Carcinoma, Non-Small-Cell Lung ; immunology ; pathology ; Female ; Humans ; Lung ; immunology ; Lung Neoplasms ; immunology ; pathology ; Male ; Middle Aged ; Neoplasm Staging ; T-Lymphocyte Subsets ; immunology ; T-Lymphocytes, Regulatory ; immunology

OBJECTIVETo evaluate whether neutrophil-lymphocyte ratio(NLR) predicts risk of recurrence in patients with advanced colon cancer undergoing curative resection followed by adjuvant chemotherapy.

METHODSA total of 149 patients with advanced colon cancer undergoing curative resection followed by adjuvant chemotherapy(FOLFOX6 protocol) were included. NLR was calculated preoperatively and before chemotherapy. The changes in NLR and the predictive value of NLR for prognosis were analyzed.

RESULTSThe NLR of 149 patients was 2.8±1.5. NLR of 3.5 was identified according to the ROC curve. NLR<3.5 and NLR≥3.5 were classified as low and high NLR group, respectively. The 5-year recurrence-free survival(RFS) of patients with high preoperative NLR(n=22) was significantly worse than that of those with low preoperative NLR(n=127)(50.9% vs. 76.4%, P=0.025). The difference of 5-year RFS between high pre-chemotherapy NLR group(n=34) and low pre-chemotherapy NLR group(n=115) was statistically significant(50.1% vs. 71.4%, P=0.032). The 5-year RFS was 79.5% in patients with low preoperative NLR converting to high pre-chemotherapy NLR(n=16), similar to the group with high pre-chemotherapy group(P=0.077). The 5-year RFS was 17.7% in patients with high preoperative NLR reverting to low pre-chemotherapy NLR(n=12), similar to the group with low pre-chemotherapy group(P=0.978). There was significant difference in 5-year RFS between the postoperatively elevated group and postoperatively decreased group(P=0.036).

CONCLUSIONAn elevated blood NLR may be a biomarker of poor RFS in patients with advanced colon cancer after curative resection and chemotherapy.

Adult ; Aged ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Chemotherapy, Adjuvant ; Colonic Neoplasms ; blood ; therapy ; Disease-Free Survival ; Female ; Fluorouracil ; therapeutic use ; Humans ; Leucovorin ; therapeutic use ; Lymphocytes ; immunology ; Male ; Middle Aged ; Neutrophils ; immunology ; Organoplatinum Compounds ; therapeutic use ; Prognosis