Molecular cloning, structure and expressional profiles of two novel single-exon genes (PoCCR6A and PoCCR6B) in the Japanese flounder (Paralichthys olivaceus).

CCR6 is an important binding receptor of CCL20 and beta-defensins, and has multiple functions in the innate and acquired immune responses. In this study, we cloned the PoCCR6A and PoCCR6B genes of the Japanese flounder and studied the gene structure and expression patterns of these two genes in bacterial infection. The full-length PoCCR6A cDNA is 1415 bp and the open reading frame (ORF) is 1113 bp, encoding a 370-amino-acid peptide. The full-length PoCCR6B cDNA is 2193 bp and the ORF is 1029 bp, encoding a 363-amino-acid peptide. The structures of PoCCR6A and PoCCR6B indicate that they are single-exon genes. The predicted proteins encoded by PoCCR6A and PoCCR6B have the typical G-protein-coupled receptor (GPCR) family signature of seven transmembrane domains and several conserved structural features. A tissue distribution analysis showed that PoCCR6A is predominately expressed in the intestine, gill, and blood, and PoCCR6B in the gill, spleen, and liver. The expression patterns of the two chemokine receptors were analyzed during bacterial infection. In spleen and kidney, the expression of PoCCR6A was significantly upregulated at 24 h after infection, whereas the expression of PoCCR6B was steady at these time points. While in intestine, both of them were upregulated at 6 h-12 h after infection, and in gill the expression levels of them were upregulated at 24 h. The patterns of expression suggested that PoCCR6A and PoCCR6B play an important role in the immune response of the Japanese flounder, especially in the mucosal tissues.

[Study on Salt Tolerance of Echinacea purpurea].

OBJECTIVE: To explore the salt tolerance of Echiancea purpurea and its mechanism. METHODS: Echiancea purpurea was used as test material in this study and six salinity levels (0, 30, 60, 90, 120 and 150 mmol/L NaCl) were set. Effects on seed germination and salt tolerance relevant physiological and biochemical indexes of Echiancea purpurea were studied. RESULTS: Salt stress suppressed the germination of Echiancea purpurea seeds, induced osmotic adjustment substances proline, soluble sugar and K+ to increase, and activities of POD and SOD to rise, and meanwhile resulted in accumulation of Na+ and decrease of K+/Na+. CONCLUSION: Echiancea purpurea can tolerant salt stress to a certain degree, but in case of high salt concentrations, severe salt injury would remarkably suppress the growth of Echinacea purpurea.

Serum high-density lipoprotein cholesterol levels predict early recurrence and prognosis of intrahepatic cholangiocarcinoma after surgical resection.

Introduction: Dysregulation in lipid metabolism contributes to the occurrence and development of various cancers. The connection between changes in lipid metabolism and the development of intrahepatic cholangiocarcinoma remains uncertain. Our objective was to investigate the significance of blood lipid levels in patients with intrahepatic cholangiocarcinoma who have undergone surgery. Methods: Ninety-seven ICC patients who underwent surgery were retrospectively enrolled. After 92.2 months of follow-up, the Kaplan-Meier analysis and Cox proportional hazard model were used to calculate overall survival and recurrence-free survival. Results: The median age of this cohort was 56 years, and 79 (81.4 %) of them were male. Eighty-eight (90.7 %) patients presented with tumor recurrence and 73 (75.3 %) died. In multivariate analyses, high-density lipoprotein cholesterol level (<0.91 vs. ≥ 0.91 mmol/L, hazard ratio [HR] = 2.55; 95 % CI: 1.38-4.71), lymph node metastasis (Yes vs. No, HR = 2.58; 95 % CI: 1.28-5.19), etiology factor (chronic HBV infection vs. others, HR = 0.5; 95 % CI: 0.28-0.88) and multiple tumor lesions (Yes vs. No, HR = 1.85; 95 % CI: 1.01-3.39) were independent predictors of overall survival. However, only high-density lipoprotein cholesterol level (HR = 1.86; 95 % CI: 1.19-2.92) emerged as the independent factor for recurrence-free survival. High-density lipoprotein cholesterol level (HR = 2.07; 95 % CI: 1.26-3.41), etiology factor (HR = 0.49; 95 % CI: 0.29-0.84), and multiple tumor lesions (HR = 2.00; 95 % CI: 1.14-3.51) were independent predictors of early recurrence. For patients who did not experience the spread of cancer to the lymph nodes, there was a significant correlation between the level of high-density lipoprotein cholesterol and their overall survival, recurrence-free survival, and early recurrence. For patients with low pre-operation high-density lipoprotein cholesterol levels, high post-operation high-density lipoprotein cholesterol levels were associated with better prognosis. Conclusions: Low serum high-density lipoprotein cholesterol level might serve as a sign of poor clinical outcomes (overall survival and recurrence-free survival) and early recurrence among intrahepatic cholangiocarcinoma patients. Strengthening the monitoring and intervention of intrahepatic cholangiocarcinoma patients with poor prognosis might be critical for improving the prognosis.

Increased Prevalence of Thyroid Nodules Across Nearly 10 Years in Shanghai, China.

OBJECTIVE: This study aimed to determine whether the prevalence of thyroid nodules (TNs) increased due to modern lifestyles or other factors, despite the advances in screening and diagnostic tools. METHODS: This study included 3474 pairs of participants, who were matched by gender and age (±3 years) from two cross-sectional sampling surveys: (1) the program on the iodine nutritional status and related health status of residents in Shanghai in 2009; (2) the thyroid disease screening program for adults in Shanghai between 2017 and 2018. The prevalence of TNs and thyroid diseases in 2009 and 2017-2018 were compared, and the potential risk factors of TNs were detected. RESULTS: The prevalence of TNs in 2009 was 28.9%: 22.5% in males and 34.5% in females. In 2017, this increased to 43.8%: 37.9% in males and 49.1% in females. The prevalence of TNs significantly increased from 2009 to 2017 (odds ratio, 1.486; 95% confidence interval, 1.238-1.786). In addition, female gender, thyroid disease history, and age were the main risk factors for TNs after adjusting for confounders in the logistic regression across the time period. CONCLUSION: The prevalence of TNs significantly increased across nearly 10 years in Shanghai.

[Study on the coumarin glucosides of Angelica dahurica].

OBJECTIVE: To study the coumarin glucosides of Angelica dahurica. METHODS: Fresh roots and rhizomes of Angelica dahurica were extracted with ethanol at room temperature. Repeated column chromatography and preparative HPLC were used to isolate and purify the compounds. Their structures were elucidated on the basis of chemical evidence and spectral analysis. RESULTS: Twenty-six coumarin glucosides were isolated from Angelica dahurica, while here we reported 7 of them: sec.-O-beta-D-Galactopyranosyl-(R)-byakangelicin (I);8-O-beta-D-Galactopyranosyl-xanthotoxol (II); 7-O-beta-D-Apiofuranosyl-(1-->6)-beta-D-Glucopyranosyl-peucedanol (III); (R)-peucedanol-7-O-beta-D-Glucopyranoside (IV); sec.-O-beta-D-Glucopyranosyl-(R)-Oxypeucedaninhydrate (V); 7-O-beta-D-Galactopyranosyl-Sco-poletin( VI); Aesculin (VII). CONCLUSION: Compound V is a new compound, Compound VII is isolated from Umbellifera for the first time, compounds III, IV are isolated from Angelica for the first time,while compound I, II and VI are isolated from this plant for the first time.

Genetic diversity and identification of Chinese-grown pecan using ISSR and SSR markers.

Pecan is an important horticultural nut crop originally from North America and now widely cultivated in China for its high ecological, ornamental and economic value. Currently, there are over one hundred cultivars grown in China, including introduced American cultivars and Chinese seedling breeding cultivars. Molecular markers were used to assess the genetic diversity of these cultivars and to identify the pedigrees of fine pecan plants with good characteristics and no cultivar-related data. A total of 77 samples grown in China were studied, including 14 introduced cultivars, 12 domestic seedling breeding cultivars, and 49 fine pecan plants with no cultivar data, together with Carya cathayensis and Juglans nigra. A total of 77 ISSR and 19 SSR primers were prescreened; 10 ISSR and eight SSR primers were selected, yielding a total of 94 amplified bands (100% polymorphic) in the range of 140-1,950 bp for the ISSR and 70 amplified bands (100% polymorphic) in the range of 50-350 bp for SSR markers. Genetic diversity analyses indicated Chinese-grown pecan cultivars and fine plants had significant diversity at the DNA level. The dengrograms constructed with ISSR, SSR or combined data were very similar, but showed very weak grouping association with morphological characters. However, the progeny were always grouped with the parents. The great diversity found among the Chinese cultivars and the interesting germplasm of the fine pecan plants analyzed in this study are very useful for increasing the diversity of the pecan gene pool. All 77 accessions in this study could be separated based on the ISSR and SSR fingerprints produced by one or more primers. The results of our study also showed that ISSR and SSR techniques were both suitable for genetic diversity analyses and the identification of pecan resources.

Characterization of the complete chloroplast genome of Carya hunanensis W. C. Cheng & R. H. Chang.

Carya hunanensis W. C. Cheng & R. H. Chang is a tree of great economic importance in China. In this study, the complete chloroplast genome of C. hunanensis was determined and analyzed phylogenetically. The whole genome was determined to be 159,892 bp in length, presenting a typical quadripartite structure with a large single copy (LSC) region (89,466 bp), a small single copy (SSC) region (18,714 bp), and a pair of inverted repeat (IR) regions (25,856 bp each). There were 132 genes annotated in the genome, including 84 protein-coding genes, 40 tRNA genes, and eight rRNA genes. The overall GC content of C. hunanensis chloroplast genome was 36.24%. Phylogenetic analysis indicated that C. hunanensis was closely related to C. kweichowensis and Annamocarya sinensis.

Characterization of the complete chloroplast genome of Carya laciniosa (F.Michx.) G.Don (Juglandaceae).

Carya laciniosa (F.Michx.) G.Don is a slow-growing valuable tree in the family Juglandaceae. In this research, we first reported the chloroplast genome sequence of C. laciniosa. The chloroplast genome was a circular form with 160,832 bp in size, comprising four subregions: a large single-copy (LSC) region of 90,065 bp, a small single-copy (SSC) region of 18,791 bp, and two copies of inverted repeats (IRs; IRa and IRb) of 25,988 bp each. A total of 132 genes were annotated in the chloroplast genome of C. laciniosa, including 87 protein-coding genes, 37 tRNA genes, and 8 rRNA genes. The overall GC content of the whole genome was 36.14%. Phylogenetic analysis suggested that C. laciniosa was closely related to C. ovata.

Occupational Health Risk Assessment of Benzene, Toluene, and Xylene in Shanghai.

OBJECTIVE: This study was designed to conduct a retrospective and systematic occupational health risk assessment (OHRA) of enterprises that used benzene, toluene, and xylene (BTX) in Shanghai, China. METHODS: All data for the study were obtained from 1,705 occupational health examination and evaluation reports from 2013 to 2017, and a semiquantitative model following Chinese OHRA guidelines (GBZ/T 298-2017) was applied for the assessment. RESULTS: The selected enterprises using BTX were mainly involved in manufacturing of products. Using the exposure level method, health risk levels associated with exposure to BTX were classified as medium, negligible, or low. However, the risk levels associated with benzene and toluene were significantly different according to job types, with gluers and inkers exhibiting greater health risks. For the same job type, the health risk levels assessed using the comprehensive index method were higher than those using the exposure level method. CONCLUSION: Our OHRA reveals that workers who are exposed to BTX still face excessive health risk. Additionally, the risk level varied depending on job categories and exposure to specific chemicals. Therefore, additional control measures recommended by OHRA guidelines are essential to reduce worker exposure levels.

[Studies on the chemical constituents of Lonicera macranthoides].

OBJECTIVE: To study the chemical constituents of flower buds of Lonicera macranthoides. METHODS: The 90% EtOH extract of Lonicera macranthoides. was successively partitioned with petroleum ether and ethyl acetete. RESULTS: Repeated column chromatography of the ethyl acetete fraction afforded the following compounds (1-9): ginnol (1), triacontanol (2), ursolic acid (3), beta-sitosterol (4), triacontane (5), palmitic acid (6), beta-daucosterol (7), 3-decyl-3-octyldocosan-1-ol (8), 3-dodecyl-3-nonyldocosan-1-ol (9). All compounds except 4 are isolated from this plant for the first time while compounds 2, 3, 5, 8 and 9 are their first time been isolated from genus Lonicera.

Characteristics of soil phosphorus fractions under different thinning intensities in Larix principis-rupprechtii plantation and the affecting factors.

Thinning is an important plantation management strategy. Phosphorus (P) is one of the limiting nutrients in forest ecosystems. The impacts of thinning on soil P remain unclear. In this study, we investigated the characteristics of soil P fractions in topsoil (0-10 cm) of Larix principis-rupprechtii plantation in Taiyue Mountain, Shanxi Province under different thinning intensity (control, CK, 0%; low thinning, LT, 15%; moderate thinning, MT, 35%; high thinning, HT, 50%) and the affecting factors. The soil P fractions were measured using Tissen modified Hedley P fractionation method. The results showed that the total inorganic P content in soil was significantly higher in stands subjected to moderate thinning than in control. The soil acid phosphatase activity (APA) and the contents of Resin-Pi, NaHCO3-Pi, NaHCO3-Po, NaOH-Pi, and microbial biomass phosphorus (MBP) were significantly increased in LT and MT compared with CK, but the content of NaOH-Po showed an opposite trend. Thinning had no significant effect on soil total P, total organic P, non-available P and residual-P. Soil moisture, organic matter, MBP and APA were the important factors affecting soil P availability. Our results demonstrated that MT could enhance soil P availability in Larix principis-rupprechtii plantation.

[Preliminary study of serum proteome biomarkers of hypopharyngeal squamous cell carcinoma].

OBJECTIVE: To screen the serum proteome biomarkers of hypopharyngeal squamous cell carcinoma (HSCC) and to establish a predictive model for early detection of HSCC. METHODS: Serum samples were collected from 48 HSCC patients before surgery and 52 age and sex-matched individuals without cancer used as controls. The samples were divided into 2 sets: training set (including 36 HSCC patients and 36 controls) and blind testing set (including 12 HSCC patients and 16 controls). With WCX2 and IMAC3 protein chips, surface-enhanced laser desorption/ionization (SELDI) was used to analyze the serum protein profiling. 72 samples of the training set were analyzed by a decision tree algorithm to be able to differentiate HSCC patients from controls. Double-blind test was used to determine the sensitivity and specificity of the classification model. RESULTS: Ranging from 2000 - 50000 (M/Z), 11 potential biomarkers on WCX2 and 19 biomarkers on IMAC3 protein chip could differentiate HSCC patients from the control set (P < 10(-5)). Among them 4 candidate protein peaks with the m/z values of 7796, 4216, 5927, and 5361 were selected to be used to establish a predictive model by Biomarker Pattern Software. The model separated effectively the HSCC samples from the control samples, achieving a sensitivity of 94.44%, and a specificity of 88.89%. An accuracy of 85.71% (24/28), sensitivity of 91.67% (11/12), specificity of 81.25% (13/16), positive predictive value of 78.57%% (11/14), and negative predictive value of 92.85% (13/14) were validated in the double-blind testing set. CONCLUSION: The SELDI-TOF-MS Protein Chip combined with artificial intelligence classification algorithm helps find serum proteome biomarkers and establish predictive model for early diagnosis of HSCC. This technique has potential for the development of a screening test for the detection of HSCC.

Restorative effects of zinc and selenium on cadmium-induced kidney oxidative damage in rats.

OBJECTIVE: To investigate whether cadmium-induced oxidative stress in the kidney is influenced by zinc and selenium. METHODS: Five groups of rats were maintained: (A) Cd (CdCl2, 400 micrograms.kg-1.day-1 intraperitoneal injection); (B) Cd + Zn (ZnCl2, 20 mg.kg-1.day-1 hypodermic injection); (C) Cd + Se (Na2SeO3, 350 micrograms.kg-1.day-1 via a stomach tube); (D) Cd + Zn + Se; (E) treated with physiological saline as a sham-handled control. The rats were given treatment for a period of 4 weeks. The activities of superoxide dismutase (SOD), glutathione peroxidase (GH-Px), catalase (CAT), and the level of malondialdehyde (MDA) in the kidney tissue were measured to assess the oxidative stress. Urinary lactate dehydrogenase (LDH) activity was used as an indicator of tubular cell damage caused by lipid peroxidation. RESULTS: In group C and D, activities of SOD (110.5 +/- 5.2, 126.8 +/- 7.0; P < 0.05) and GSH-Px (85.7 +/- 4.9, 94.6 +/- 7.3; P < 0.05) were higher than those in group A (84.7 +/- 3.3; 56.9 +/- 3.8); and in group B, only the activity of GSH-Px (80.0 +/- 4.3, P < 0.01) increased in comparison with that in group A (56.9 +/- 3.8). Significant increase of MDA (P < 0.05) was seen in group B (31.1 +/- 4.7) and C (35.0 +/- 4.1) when compared with control values (17.2 +/- 1.8). No difference was found in the level of MDA between group D (18.9 +/- 2.6) and control. The activity of LDH in urine of control group (0.06 +/- 0.02) was lower than that of group A (0.46 +/- 0.19, P < 0.05), B (0.10 +/- 0.05, P < 0.05) and C (0.14 +/- 0.07, P < 0.05), and there was no significant change between control (0.06 +/- 0.02) and group D (0.08 +/- 0.02). CONCLUSION: Zinc or selenium could partially alleviate the oxidative stress induced by cadmium in kidney, but administration cadmium in combination with zinc and selenium efficiently protects kidney from cadmium-induced oxidative damage.

Synthesis of beta-aminoketones and construction of highly substituted 4-piperidones by mannich reaction induced by persistent radical cation salts.

A Mannich reaction of imines and ketones induced by persistent radical cation salts was investigated, and a series of Mannich bases, beta-aminoketones, were synthesized. A novel cyclization to form the 4-piperidone skeleton was achieved in a tandem process. The reaction can be rationalized as a radical cation process supported by various evidence.