[The expressions of CTLA-4 and PD-1 on CD(4)(+) T cells and the level of plasma VEGF in patients with obstructive sleep apnea hypopnea syndrome].

Objective: CD(4)(+)T cells, cytotoxic T-lymphocyte antigen 4 (CTLA-4), programmed cell death-1 (PD-1) and vascular endothelial growth factor (VEGF) are associated with cancer development. The aim of the present study was to investigate the expression of CTLA-4, PD-1 and VEGF in patients with obstructive sleep apnea hypopnea syndrome (OSAHS). Methods: From January 2017 to January 2018, a total of 47 first-visit outpatients were recruited in the Sleep and Respiratory Disorder Center of Guangdong Provincial People's Hospital, and were divided into control group (N=17, mean age 54±12 years), mild-to-moderate OSAHS group (N=15, mean age 54±12 years) and severe OSAHS group (N=15, mean age 56±13 years). Venous blood was collected, plasma and cells were isolated, the expressions of PD-1 and CTLA-4 on the surface of CD(4)(+)T cells were detected by flow cytometry, and plasma VEGF was measured by enzyme linked immunosorbent assay. Results: The proportion of CD(4)(+)T cells in control group, mild-to-moderate OSAHS group and severe OSAHS group were respectively(38±8)%, (35±8)% and (38±6)% (F=1.228, P>0.05). The expression of CTLA-4 on CD(4)(+)T cells were respectively [1.13 (0.59~1.78)]%, [0.45 (0.16~1.43)]% and [0.87(0.47~1.46)]% (H=2.205, P>0.05). The expression of PD-1 on CD(4)(+)T cells were respectively [4.24 (2.12~6.03)]%, [3.54(2.69~5.09)]% and [3.31(1.67~8.25)]% (H=0.541, P>0.05). The concentrations of VEGF in control group, mild-to-moderate OSAHS group and severe OSAHS group were statistically different [(395.16±87.78) ng/L vs (452.85±107.97) ng/L vs (546.42±199.27) ng/L, F=4.827, P=0.013]. Compared with the control group, VEGF concentration was significantly increased in the severe OSAHS group(P<0.01). VEGF concentration was correlated negatively with the lowest SpO(2) (r (s)=-0.480,P=0.001), but positively with apnea-hypopnea index(r (s)=0.403, P=0.005), oxygen desaturation index (r (s)=0.378, P=0.010) and proportion of SpO(2) less than or equal to 90% of total sleep time(r (s)=0.547, P=0.000 3). Conclusion: There was no significant difference of PD-1 and CTLA-4 expression on CD(4)(+)T cells in patients with and without OSAHS. The expression of VEGF was elevated in OSAHS patients, and increased with the severity of OSAHS and hypoxia.

[The effect on myocardial perfusion and clinical outcome of intracoronary nicorandil injection prior to percutaneous coronary intervention in ST-segment elevation myocardial infarction].

Objective: To investigate the effect of intracoronary administration of nicorandil prior to primary percutaneous coronary intervention (PPCI) on myocardial perfusion and short-term clinical outcomes in patients with ST-segment elevation myocardial infarction (STEMI). Methods: A total of 158 patients with STEMI undergoing PPCI from January 2014 to December 2015 in Fuzhou General Hospital were enrolled consecutively in this prospective controlled randomized trial. Patients were assigned into three groups with random number table: the nicorandil group (patients received intracoronary administration of 6 mg nicorandil after guide wire or balloon successfully crossed the target lesion, n=53), the nitroglycerin group (patients received intracoronary administration of 300 µg nitroglycerin after after guide wire or balloon successfully crossed the target lesion, n=52) and the control group(patients received routine treatment, n=53). The primary outcomes were myocardial perfusion, including the levels of corrected TIMI frame count (cTFC), and the incidence of no reflow or slow flow after PPCI. The secondary outcomes included the incidence of major adverse cardiovascular events (MACE) during hospitalization (all-cause death, reperfusion arrhythmia within 2 hours after PPCI, angina within 24 hours after PPCI, new heart failure or worsening cardiac function, and repeat revascularization) and within 3 months of follow-up (all-cause death, nonfatal myocardial infarction, repeat revascularization, post-infarction angina, and re-hospitalization for congestive heart failure). Results: The age of enrolled patients was (62.9±11.3) years old, and 130 cases (82.3%) of them were male. The median time of symptom-onset to balloon was 4.50 (3.20, 6.43) hours. There were significantly difference in cTFC immediately after PPCI((21.68±7.43)frames, (24.74±8.66)frames, and(27.06±10.40)frames), incidence of no reflow or slow flow after PPCI(5.7%(3/53), 13.5%(7/52), and 22.6%(12/53)), ST-segment resolution at 2 hours after procedure(90.6%(48/53), 84.6%(44/52), and 74.5%(38/53)), and reperfusion arrhythmia at 2 hours after procedure(15.1%(8/53), 36.6%(19/52), and 34.0%(18/53)) among the 3 groups(P<0.01 or 0.05). In the multivariate logistic regression models, intracoronary administration of nicorandil could lower the cTFC level (OR=0.17, 95%CI 0.10-0.41, P=0.001), acted as a protecting factor on lowering the incidence of no reflow or slow flow (OR=0.13, 95%CI 0.02-0.96, P=0.045) and reperfusion arrhythmia (OR=0.26, 95%CI 0.09-0.74, P=0.012), as well as facilitating the ST-segment resolution at 2 hours after procedure (OR=4.62, 95%CI 1.14-18.82, P=0.033). However, observed parameters were similar between intracoronary administration of nitroglycerin group compared with control group (all P>0.05). MACE within 3 months of follow-up were similar among the 3 groups(all P>0.05). Conclusion: Intracoronary administration of nicorandil prior to balloon dilation can significantly improve the myocardial perfusion and reduce the occurrence of reperfusion arrhythmia during PPCI for STEMI, but does not affect the short-term prognosis in STEMI patients.

Effect of atorvastatin on vascular endothelial function in moderately nicotine-dependent smokers.

We investigated the effect of atorvastatin on vascular endothelial function in moderately nicotine-dependent smokers. One hundred and sixty moderately nicotine-dependent smokers were randomly divided into the atorvastatin group (N = 80) and the control group (N = 80). Total cholesterol (TC), triglyceride, high-density lipoprotein cholesterol, low-density lipoprotein cholesterol (LDL-C), fasting plasma glucose levels, aspartate aminotransferase, total bilirubin, creatine phosphokinase, and brachial artery flow-mediated vasodilation function (FMD) were measured before and 12 months after atorvastatin treatment. After a 12-month atorvastatin therapy, the TC and LDL-C levels of patients were decreased significantly (P < 0.05) and the FMD of patients were improved significantly (P < 0.05). Compared with the control group, TC and LDL-C of the patients were significantly decreased (P < 0.05) and the FMD of the patients were significantly improved (P < 0.05). Atorvastatin may significantly improve endothelial function in moderately nicotine-dependent smokers.

Short communication: development and characterization of novel transcriptome-derived microsatellites for genetic analysis of persimmon.

Oriental persimmon (Diospyros kaki Thunb.) (2n = 6x = 90) is a major commercial and deciduous fruit tree that is believed to have originated in China. However, rare transcriptomic and genomic information on persimmon is available. Using Roche 454 sequencing technology, the transcriptome from RNA of the flowers of D. kaki was analyzed. A total of 1,250,893 reads were generated and 83,898 unigenes were assembled. A total of 42,711 SSR loci were identified from 23,494 unigenes and 289 polymerase chain reaction primer pairs were designed. Of these 289 primers, 155 (53.6%) showed robust PCR amplification and 98 revealed polymorphism between 15 persimmon genotypes, indicating a polymorphic rate of 63.23% of the productive primers for characterization and genotyping of the genus Diospyros. Transcriptome sequence data generated from next-generation sequencing technology to identify microsatellite loci appears to be rapid and cost-efficient, particularly for species with no genomic sequence information available.

Automated 3D segmentation of hippocampus based on active appearance model of brain MR images for the early diagnosis of Alzheimer's disease.

AIM: To investigate the hippocampal regional deformation modes by means of a novel method of automatic segmentation for discriminating between Alzheimer's disease (AD) and normal aging; and to further provide the effective evidence for the early diagnosis of AD. METHODS: Twenty AD patients and sixty healthy volunteers were included in this retrospective study. High-resolution structural volumetric images were obtained on a 3.0 T MR imaging system. Data were processed to create three-dimensional (3D) active appearance model (AAM) of hippocampus. Automatic recognition and 3D segmentation were carried out on both sides of the hippocampus in brain MR images of individuals with this model, and the hippocampal statistical shape model was established for AD group and control group. Student's t test was used to identify whether there was difference between AD group and control group in the hippocampal regional deformation detected by automatic segmentation, and to compare whether there was difference between the automated segmentation and the manual tracing for quantifying hippocampal volumes on left/right side of the same sex group of healthy volunteers and if there was genderwise difference. Pearson's Correlation test was employed to determine whether there was a correlation between automated segmentation and manual tracing for quantifying hippocampal volumes. RESULTS: No significant difference was detected between automated segmentation and manual tracing for quantifying hippocampal volumes on left/right side of the same sex group of healthy volunteers (P>0.05). Further there was no significant genderwise difference (P>0.05). A very strong positive correlation existed between both methods for quantifying hippocampal volumes (denoted R(2) near 1.0, P<0.001). Noticeable atrophy of bilateral hippocampal head was found among twenty patients with AD through statistical shape model compared with control group (P<0.05), especially on the left where inward-deformation was significantly found. CONCLUSION: This novel method of automated segmentation of the hippocampus based on AAM has been found to be reliable and accurate in our study, which may be an alternative to manual segmentation. The featured atrophy of hippocampal head can be regarded as an important biomarker for the early diagnosis of AD.

Comparison of dual-source parallel radio frequency transmission liver MRI at 3.0 T with conventional MRI.

AIM: Aim of the present study was to investigate the role of dual-source parallel Radio frequency (RF) and single-source excitation in liver imaging at 3.0 T MR. METHODS: This study was a retrospective analysis. One hundred and seven subjects underwent a 3.0 T TX MR scanning including axial spectrally selective attenuated inversion recovery (SPAIR) T2WI, axial DWI and coronal balanced-fast field echo (Balanced FFE). Each sequence was carried out with both single-source and dual-source RF excitation. Student's t test was used to compare whether there was difference between single-source and dual-source RF excitation in the image uniformity, single-to-noise ratio (SNR) and contrast-to-noise ratio (CNR). Wilcoxon signed rank test was used to determine whether there was difference between conventional and parallel transmission in the score of image quality. Reader agreement was assessed using the Cohen's Kappa test. RESULTS: For the image uniformity, there was significant difference between single-source and dual-source excitation (418.40±66.75 for single-source vs. 416.26±50.61 for dual-source, t=2.524, P<0.05). There also existed significant difference between single-source and dual-source excitation in SNR and CNR, respectively. The SNR and CNR of parallel transmission (22.03±12.16 and 18.33±10.01, respectively) were both higher than those of single transmission (20.36±11.21 and 15.22±8.95, respectively) (t=-2.630, P<0.05 for SNR and t=-4.238, P<0.05 for CNR). Image quality comparisons revealed significantly better results with dual-source than single-source RF excitation at SPAIR T2WI (1.4±0.42 vs. 1.81±0.27), DWI (1.08±0.46 vs. 1.63±0.36) and balanced FFE sequence (0.95±0.45 vs. 1.65±0.37, Z=-5.894, -5.801 and -6.985, respectively, P<0.01). In the comparison of image quality, the agreement between the two readers was very good (Kappa>0.8, P<0.05). CONCLUSION: Dual-source parallel RF excitation MR imaging in liver enables reducing dielectric shading, improving homogeneity of the RF magnetic induction field, and increasing SNR and CNR at 3.0 T.

Transferability of retrotransposon primers derived from Persimmon (Diospyros kaki Thunb.) across other plant species.

Retrotransposon-based molecular markers are powerful molecular tools. However, these markers are not readily available due to the difficulty in obtaining species-specific retrotransposon primers. Although recent techniques enabling the rapid isolation of retrotransposon sequences have facilitated primer development, this process nonetheless remains time-consuming and costly. Therefore, research into the transferability of retrotransposon primers developed from one plant species onto others would be of great value. The present study investigated the transferability of retrotransposon primers derived from 'Luotian-tianshi' persimmon (Diospyros kaki Thunb.) across other fruit crops, as well as within the genus using inter-retrotransposon amplified polymorphism molecular marker. Fourteen of the 26 retrotransposon primers tested (53.85%) produced robust and reproducible amplification products across all fruit crops tested, indicating their applicability across plant species. Four of the 13 fruit crops showed the best transferability performances: persimmon, grape, citrus, and peach. Furthermore, similarity coefficients and UPGMA clustering indicated that these primers could further offer a potential tool for germplasm differentiation, parentage identification, genetic diversity assessment, classification, and phylogenetic studies across a variety of plant species. Transferability was further confirmed by examining published primers derived from Rosaceae, Gramineae, and Solanaceae. This study is one of the few currently available studies concerning the transferability of retrotransposon primers across plant species in general, and is the first successful study of the transferability of retrotransposon primers derived from persimmon. The primers presented here will help reduce costs for future retrotransposon primer development and therefore contribute to the popularization of retrotransposon molecular markers.

Characterization and comparison of EST-SSR and TRAP markers for genetic analysis of the Japanese persimmon Diospyros kaki.

We developed and characterized expressed sequence tags (ESTs)-simple sequence repeats (SSRs) and targeted region amplified polymorphism (TRAP) markers to examine genetic relationships in the persimmon genus Diospyros gene pool. In total, we characterized 14 EST-SSR primer pairs and 36 TRAP primer combinations, which were amplified across 20 germplasms of 4 species in the genus Diospyros. We used various genetic parameters, including effective multiplex ratio (EMR), diversity index (DI), and marker index (MI), to test the utility of these markers. TRAP markers gave higher EMR (24.85) but lower DI (0.33), compared to EST-SSRs (EMR = 3.65, DI = 0.34). TRAP gave a very high MI (8.08), which was about 8 times than the MI of EST-SSR (1.25). These markers were utilized for phylogenetic inference of 20 genotypes of Diospyros kaki Thunb. and allied species, with a result that all kaki genotypes clustered closely and 3 allied species formed an independent group. These markers could be further exploited for large-scale genetic relationship inference.

Cryopreservation of in vitro grown shoot tips of Diospyros kaki thunb. using different methods.

The objective of this study was to compare the potential of different cryopreservation strategies for in vitro shoot tips of Diospyros kaki Thunb. The treatments consisted of three different cryopreservation methods: vitrification, droplet-vitrification and modified droplet-vitrification. The following variables were assessed: cold acclimation, sucrose concentration in the preculture medium and PVS2 treatment time. A higher average survival level was obtained using the modified droplet-vitrification method compared to the other two methods.

Genetic relationships of the Japanese persimmon Diospyros kaki (Ebenaceae) and related species revealed by SSR analysis.

Simple sequence repeat (SSR) molecular markers based on 18 primers were employed to study the genetic relationship of Japanese persimmon (Diospyros kaki) specimens. Two hundred and sixty-two bands were detected in 30 Japanese persimmon samples, including 14 Japanese and 10 Chinese genotypes of Japanese persimmon (Diospyros kaki) and six related species, D. lotus, D. glaucifolia, D. oleifera, D. rhombifolia, D. virginiana, and Jinzaoshi (unclassified - previously indicated to be D. kaki). All SSR primers developed from D. kaki were successfully employed to reveal the polymorphism in other species of Diospyros. Most of the primers were highly polymorphic, with a degree of polymorphism equal to or higher than 0.66. The results from the neighbor-joining dendrogram and the principal coordinate analysis diagram were the same; i.e., the Chinese and Japanese genotypes and related species were separated and the relationships revealed were consistent with the known pedigrees. We also concluded that 'Xiangxitianshi' from Xiangxi municipality, Hunan Province, China, is actually a sport or somaclonal variant of 'Maekawa-Jirou', and that 'Jinzaoshi' should be classified as a distinct species of Diospyros. We found that SSR markers are a valuable tool for the estimation of genetic diversity and divergence in Diospyros.

Use of in vivo imaging system-bioluminescence imaging to inspect tumor dynamic morphology.

AIM: The aim of this study was to evaluate the veracity and sensitivity of in-vivo imaging system (IVIS) for inspection of tumor dynamic morphology. METHODS: Mouse melanoma cells (B16-F10-luc-G5) in 100 mL media were seeded into a 96-well plate by 1:2 serial dilution from 10000 cells (well #1) to 78 cells (well #8). The plate was imaged using IVIS system to evaluate its sensitivity for luminescence. Ten Bablc mice with tumor cells were injected subcutaneously (1 x 10(5) in 100 mL) and tumor luminescence was detected by IVIS at Day 0, Day 3, Day 5, Day 7 and Day 9. RESULTS: As few as 78 tumor cells were detectable by IVIS. A strong correlation between number of tumor cells and bioluminescence (R2=0.99) was also demonstrated. Tumor luminescence were observed in all mice by IVIS at all days, and there was significant difference (P<0.01) between each two days from Day 0 to Day 9. Moreover, tumor dynamic morphology could be monitored by IVIS when it is invisible. CONCLUSION: Compared with conventional methods, with high veracity and sensitivity, IVIS system should be recommended as an effective method for inspection of tumor dynamic morphology.

[Effect of Puerarin on platelet activating factors CD63 and CD62P, plasminogen activator inhibitor and C-reactive protein in patients with unstable angia pectoris].

OBJECTIVE: To explore the action of platelet surface activity protein (platelet granule membrane protein CD63 and lysosome membrane protein CD62P), plasminogen activator inhibitor (PAI-1) and C-reactive protein (C-RP) in occurrence and development of unstable angina pectoris (UAP), and the effect of Puerarin on them. METHODS: Patients with UAP were randomly divided into the treated group (32 cases, treated with Puerarin) and the control group (27 cases, treated with Ticlid), the therapeutic course was 4 weeks for both groups. Changes of CD63, CD62P, PAI-1 and C-RP before and after treatment were observed. RESULTS: The levels of CD63, CD62P, PAI-1 and C-RP were higher in UAP patients than those in normal subjects and in the patients with stable angina pectoris (P < 0.05 or P < 0.01). These parameters were increased along with the aggravating of patients in Braunwald's degree. After 4-week treatment, the above-mentioned parameters lowered in both groups (P < 0.05 or P < 0.01), and comparison between the two groups showed no significant difference statistically. CONCLUSION: Platelet activation, plasminogen activator and C-RP play important roles in the occurrence and development of UAP. The obvious effect of Puerarin in anti-platelet activation, improving plasminogen activator and relieving inflammatory reaction was of great importance in preventing the occurrence and development of acute coronary syndrome clinically.

Cardiac troponin I: a marker for post-burn cardiac injury.

Cardiac troponin I (cTnI) was measured by chemiluminescent immunoassay following burn injury. Thirty patients [total body surface area (TBSA) of burn 15-98%] were included in this study and each had four to six blood samples collected at 2-day intervals between the 5th and 14th days post-burn. All patients were found to have increased cTnI on two or more occasions. The mean cTnI concentration was significantly higher in patients with TBSA of burn > 30% (0.34 microg/L compared with 0.09 microg/L, P<0.001) and in those with obvious burn wound exudation (0.32 microg/L compared with 0.12 microg/L, P<0.01). cTnI concentrations peaked at the time when there was obvious burn wound exudation or spontaneous separation of eschar, then decreased after surgical excision. Two patients with persistently high cTnI concentrations developed tachycardia. We conclude that burned patients have varying degrees of non-ischaemic cardiac injury, manifesting as leakage of cTnI from myocytes into the circulation.

[Early diagnosis and treatment of contralateral primary glaucoma associated with neovascular glaucoma].

The early diagnosis and treatment of contralateral primary glaucoma associated with neovascular glaucoma (NVG) are discussed. Treatment of the underlying disease of NVG and long term follow-up of both eyes are emphasized in order to prevent blindness in both eyes.

Co-localization of an endocytic marker and acid phosphatase in a tubular/reticular compartment in macrophages.

Cultured resident murine macrophages are incubated in the continuous presence of the fluorescent endocytic marker Lucifer Yellow and a phorbol ester that activates protein kinase C. Under these steady-state labeling conditions the fluorescent tracer was, for the most part, in a tubular/reticular compartment. Enzyme cytochemical localization of acid phosphatase in the same cells showed essentially a one-to-one correlation between the Lucifer Yellow- and acid phosphatase-containing compartments. Procedures for epifluorescence observation and subsequent enzyme cytochemical examination of the same whole mount cells are described. In addition, chemical fixation methods for the preservation of this labile tubular/reticular compartment are presented.

Ultrastructural localization of phosphatase activity in the guinea pig pineal gland by the cerium technique.

Thiamine pyrophosphatase (TPPase), nucleoside diphosphatase (NDPase), and glucose-6-phosphatase (G-6-Pase) were localized by the cerium technique in guinea pig pinealocytes and compared with the corresponding lead technique. NDPase and TPPase were also compared at different pH values using the cerium technique. Vibratome sections of perfusion-fixed tissue were incubated with cerium chloride or lead nitrate. Substrates used were thiamine pyrophosphate (for TPPase), sodium inosine diphosphate (NDPase), and disodium glucose-6-phosphate (G-6-Pase). The 1-2 trans saccules of the Golgi apparatus showed TPPase and NDPase activity but none for G-6-Pase. The endoplasmic reticulum (ER) cisternae and perinuclear space had NDPase and G-6-Pase activity but not TPPase. The abluminal plasmalemma of endothelial cells and the plasmalemma of Schwann cells demonstrated TPPase and NDPase activity but the luminal plasmalemma of the endothelial cells and the plasmalemma of pinealocyte processes showed only NDPase activity. TPPase was active at all pH values tested, but NDPase was most active at pH values of 6.5 and 7.0. Lead phosphate precipitate was frequently seen in nuclei, perinuclear space, ER cisternae, and "synaptic" vesicles when lead was used as the capturing agent. These sites were usually not labeled when cerium was used.

Ultrastructural localization of acetylcholinesterase in the guinea pig pineal gland.

The ultrastructural localization of acetylcholinesterase (AChE) activity in guinea pig pineal gland was studied using the copper-glycine procedure. A small number of pinealocytes and bundles of unmyelinated nerve fibers were labeled by the AChE reaction. The AChE-positive pinealocytes were located near blood vessels and distributed in small groups. The AChE reaction product was localized in the perinuclear cistern, in the cisternae of the endoplasmic reticulum (ER), and in the saccules of the Golgi apparatus. These findings suggest that the AChE-positive pinealocytes synthesize AChE. The AChE reaction product was also seen in the intercellular space between pinealocyte processes. Besides pinealocytes, AChE activity was localized on the axolemma of myelinated and unmyelinated nerve fibers and in the basement membrane surrounding unmyelinated nerve fibers. Pseudocholinesterase activity was confined to Schwann cells, which showed the reaction product in their perinuclear cistern, in the cisternae of the ER, and on the plasmalemma.

Prenatal development of "synaptic" ribbons in the guinea pig pineal gland.

Pineal "synaptic" ribbons are a heterogeneous population of organelles. "Synaptic" ribbons (SR) sensu stricto, "synaptic" spherules (SS), and intermediate forms (IMF) are present. Their function and origin are unknown, and a knowledge of their prenatal development is lacking. Thus the pineal glands of prenatal, neonatal, and adult guinea pigs were prepared for electron microscopy. "Synaptic" ribbons were studied morphologically and quantitatively. The three categories of "synaptic" ribbons reported in adult pineal glands were also present in prenatal pineal glands. Their structural features, distribution, grouping, and composition patterns are similar to those in adults. "Synaptic" ribbons were first detected in pinealocytes of the distal region of a 42-day postcoitus (PC) pineal gland and were comparable with those in adults. They increased in number with age and reached a peak at 63 days PC, followed by a steep decline at 66 and 67 days PC. By day 69 PC, the numbers increased again and showed a dramatic increase after birth. Several true ribbon synapses were seen at day 63 PC between pinealocyte cell processes or between pinealocyte cell process and pinealocyte cell body. Since true ribbon synapses have not been found in adult guinea pig pinealocytes, their synaptic nature could have been lost during development. No precursors for the "synaptic" ribbons were found. The endoplasmic reticulum cisternae may be the origin for the ribbon vesicles because of their close association with the "synaptic" ribbons.