Comparison of the two histological subtypes of ampullary adenocarcinoma: a retrospective study.

OBJECTIVES: This study aimed to compare the intestinal and pancreatobiliary subtypes of ampullary adenocarcinoma in a large patient group due to limited data on survival and risk factors. METHODS: A retrospective analysis of the clinical and pathological findings and the survival of 184 patients with ampullary adenocarcinoma who underwent curative operation between 2007 and 2018 was performed. RESULTS: Pancreatobiliary subtype had a higher prevalence of jaundice before operation than the intestinal subtype (p < 0.05). Pancreatobiliary subtype had a larger tumor size (> 2 mm) (p < 0.01) and poorer differentiation (p < 0.05) than the intestinal subtype. Perineural invasion more frequently occurred in pancreatobiliary subtype than the intestinal subtype (p < 0.01) and pancreatobiliary subtype had a higher prevalence of positive dissected lymph nodes (p < 0.05) with an advanced disease stage (p < 0.01) than the intestinal subtype. Patients of the pancreatobiliary subtype had poorer disease-free and overall survival than patients of the intestinal subtype. No survival benefit of adjuvant chemotherapy was found in either patients of the intestinal subtype or pancreatobiliary subtype. No significant difference was found in any subtypes regarding the recurrent regions. CONCLUSIONS: Pancreatobiliary subtype exhibited a higher recurrence rate and a poorer overall survival rate with more unfavorable pathological characteristics than the intestinal subtype.

OBJETIVOS: Los datos sobre la supervivencia y los factores de riesgo del adenocarcinoma ampular son limitados debido a su rareza. Este estudio buscó comparar el subtipo intestinal y el subtipo pancreático-biliar en pacientes con adenocarcinoma ampular. MÉTODOS: Análisis retrospectivo de hallazgos clínicos y patológicos y la supervivencia de 184 pacientes con adenocarcinoma ampular tratados entre 2007 y 2018. RESULTADOS: El subtipo pancreático-biliar tuvo una mayor prevalencia de ictericia antes de la operación y un tamaño de tumor mayor, y una peor diferenciación, que el subtipo intestinal. La invasión perineural fue más frecuente en el subtipo pancreático-biliar, con una mayor prevalencia de linfonodos disecados positivos y un estadio avanzado de la enfermedad. Los pacientes del subtipo pancreático-biliar tuvieron una supervivencia libre de enfermedad y una supervivencia general peores que los pacientes del subtipo intestinal. No se encontró ningún beneficio de la quimioterapia adyuvante en pacientes del subtipo intestinal o pancreático-biliar. No hubo diferencia significativa en las regiones recurrentes. CONCLUSIÓN: El subtipo pancreático-biliar mostró una tasa de recurrencia y una tasa de supervivencia general peores, con características patológicas más desfavorables que el subtipo intestinal.

Roles of circ_0000135/miR-140-3p/PDZK1 network in cervical cancer.

PURPOSE: To explore the effect of circ_0000135/miR-140-3p/PDZ domain containing 1 (PDZK1) on the occurrence and development of cervical cancer. METHODS: Clinical data were collected to verify circ_0000135/miR-140-3p/PDZK1 expression in cervical cancer. mRNA expressions of circ_0000135 and miR-140-3p were detected by real-time quantitative PCR. Correlation between circ_0000135 and miR-140-3p/miR-140-3p and PDZK1 was analyzed in vitro. Protein expression detection in cells was conducted by Western blot; while cell proliferation, invasion and cycle distribution by CCK8 assay, Transwell chamber assay and flow cytometry, respectively. Rescue and animal experiment were performed to verify the effect of circ_0000135/miR-140-3p/PDZK1 on cervical cancer. RESULTS: circ_0000135 and PDZK1 expressions were increased, while those of miR-140-3p were decreased in cervical cancer tissues and cells (both P < 0.05). sh-circ_0000135 group had decreased cell viability, arrested cells in G0/G1 phase, decreased CyclinD1 expression, inhibited cell migration and invasion; sh-circ_0000135 group showed reduced tumor volume, weight, and lower Ki67 expression (all P < 0.05). circ_0000135 had conserved target of miR-140-3p. There was a direct interaction between circ_0000135 and miR-140-3p. miR-140-3p might have direct interaction with PDZK1. sh-circ_0000135 and/or miR-140-3p treatment showed obviously decreased PDZK1 expression, decreased cell activity, arrested cells in G0/G1 phase, downregulated cell migration and invasion; sh-circ_0000135 and/or miR-140-3p mimic treatment showed obviously decreased tumor volume, tumor weight, and Ki67 expression (all P < 0.05). CONCLUSION: circ_0000135 may play an anti-tumor role on the progression of cervical cancer by sponging miR-140-3p to suppress the expression of PDZK1, providing a promising therapeutic target.

Effect of oviposition by Bactrocera dorsalis on the antioxidant activity of orange juice / Efeito da oviposição de Bactrocera dorsalis na atividade antioxidante do suco de laranja

Abstract Among fruits and fruit products, oranges and orange juice are the most widely consumed worldwide. However, the effects of pest infestation of oranges on the quality of orange juice are not yet known. To evaluate the effect of the oriental fruit fly Bactrocera dorsalis on the antioxidant activity of orange juice, we measured changes in the vitamin C (Vc) concentration, total phenol content, and antioxidant activity of orange juice after the introduction of fruit fly eggs. Ten days after the eggs were introduced (larvae removed), the concentration of Vc in orange juice was 18.65 µg/mL, which was 9.16 µg/mL lower than that measured in healthy orange juice. In addition, the total phenol content decreased by 46.519 mg Gallic Acid Equivalents (GAE)/g to 9.748 mg GAE/g. Furthermore, the free-radical scavenging activity decreased from 22.297% to 5.393%. Correlation analysis indicated significant correlations between Vc concentration, total phenol content, and antioxidant activity of orange juice after B. dorsalis infestation. The decrease in Vc concentration, total phenol content and free-radical scavenging activity indicated that B. dorsalis changed the quality of orange juice by affecting the antioxidant activity of the juice after the oranges were infested.

Resumo Entre frutas e produtos de frutas, as laranjas e suco de laranja são os mais consumidos em todo o mundo. No entanto, os efeitos da infestação de laranjas sobre a qualidade do suco de laranja ainda não são conhecidos. Para avaliar o efeito da mosca-das-frutas oriental, Bactrocera dorsalis, sobre a atividade antioxidante do suco de laranja, foram medidas as mudanças na concentração de vitamina C (Vc), no teor de fenol total e na atividade antioxidante do suco de laranja após a introdução de ovos da mosca-das-frutas oriental. Dez dias após a introdução dos ovos (larvas removidas), a concentração de Vc no suco de laranja foi de 18,65 µg/mL, que foi 9,16 µg/mL menor do que a medida em suco de laranja saudável. Além disso, o teor total de fenol diminuiu de 46,519 mg em equivalente de ácido gípico (GAE)/g para 9,748 mg de GAE/g. Ademais, a atividade de eliminação de radicais livres diminuiu de 22,297% para 5,393%. A análise de correlação indicou correlações significativas dentre a concentração de Vc, o conteúdo total de fenol e a atividade antioxidante do suco de laranja após a infestação por B. dorsalis. A diminuição na concentração de Vc, o conteúdo total de fenol e a atividade sequestradora de radicais livres indicaram que B. dorsalis alterou a qualidade do suco de laranja, afetando a atividade antioxidante do suco após a infestação das laranjas.

Effect of oviposition by Bactrocera dorsalis on the antioxidant activity of orange juice.

Among fruits and fruit products, oranges and orange juice are the most widely consumed worldwide. However, the effects of pest infestation of oranges on the quality of orange juice are not yet known. To evaluate the effect of the oriental fruit fly Bactrocera dorsalis on the antioxidant activity of orange juice, we measured changes in the vitamin C (Vc) concentration, total phenol content, and antioxidant activity of orange juice after the introduction of fruit fly eggs. Ten days after the eggs were introduced (larvae removed), the concentration of Vc in orange juice was 18.65 µg/mL, which was 9.16 µg/mL lower than that measured in healthy orange juice. In addition, the total phenol content decreased by 46.519 mg Gallic Acid Equivalents (GAE)/g to 9.748 mg GAE/g. Furthermore, the free-radical scavenging activity decreased from 22.297% to 5.393%. Correlation analysis indicated significant correlations between Vc concentration, total phenol content, and antioxidant activity of orange juice after B. dorsalis infestation. The decrease in Vc concentration, total phenol content and free-radical scavenging activity indicated that B. dorsalis changed the quality of orange juice by affecting the antioxidant activity of the juice after the oranges were infested.

Effect of oviposition by Bactrocera dorsalis on the antioxidant activity of orange juice

Among fruits and fruit products, oranges and orange juice are the most widely consumed worldwide. However, the effects of pest infestation of oranges on the quality of orange juice are not yet known. To evaluate the effect of the oriental fruit fly Bactrocera dorsalis on the antioxidant activity of orange juice, we measured changes in the vitamin C (Vc) concentration, total phenol content, and antioxidant activity of orange juice after the introduction of fruit fly eggs. Ten days after the eggs were introduced (larvae removed), the concentration of Vc in orange juice was 18.65 µg/mL, which was 9.16 µg/mL lower than that measured in healthy orange juice. In addition, the total phenol content decreased by 46.519 mg Gallic Acid Equivalents (GAE)/g to 9.748 mg GAE/g. Furthermore, the free-radical scavenging activity decreased from 22.297% to 5.393%. Correlation analysis indicated significant correlations between Vc concentration, total phenol content, and antioxidant activity of orange juice after B. dorsalis infestation. The decrease in Vc concentration, total phenol content and free-radical scavenging activity indicated that B. dorsalis changed the quality of orange juice by affecting the antioxidant activity of the juice after the oranges were infested.(AU)

Entre frutas e produtos de frutas, as laranjas e suco de laranja são os mais consumidos em todo o mundo. No entanto, os efeitos da infestação de laranjas sobre a qualidade do suco de laranja ainda não são conhecidos. Para avaliar o efeito da mosca-das-frutas oriental, Bactrocera dorsalis, sobre a atividade antioxidante do suco de laranja, foram medidas as mudanças na concentração de vitamina C (Vc), no teor de fenol total e na atividade antioxidante do suco de laranja após a introdução de ovos da mosca-das-frutas oriental. Dez dias após a introdução dos ovos (larvas removidas), a concentração de Vc no suco de laranja foi de 18,65 µg/mL, que foi 9,16 µg/mL menor do que a medida em suco de laranja saudável. Além disso, o teor total de fenol diminuiu de 46,519 mg em equivalente de ácido gípico (GAE)/g para 9,748 mg de GAE/g. Ademais, a atividade de eliminação de radicais livres diminuiu de 22,297% para 5,393%. A análise de correlação indicou correlações significativas dentre a concentração de Vc, o conteúdo total de fenol e a atividade antioxidante do suco de laranja após a infestação por B. dorsalis. A diminuição na concentração de Vc, o conteúdo total de fenol e a atividade sequestradora de radicais livres indicaram que B. dorsalis alterou a qualidade do suco de laranja, afetando a atividade antioxidante do suco após a infestação das laranjas.(AU)

Effect of oviposition by Bactrocera dorsalis on the antioxidant activity of orange juice

Abstract Among fruits and fruit products, oranges and orange juice are the most widely consumed worldwide. However, the effects of pest infestation of oranges on the quality of orange juice are not yet known. To evaluate the effect of the oriental fruit fly Bactrocera dorsalis on the antioxidant activity of orange juice, we measured changes in the vitamin C (Vc) concentration, total phenol content, and antioxidant activity of orange juice after the introduction of fruit fly eggs. Ten days after the eggs were introduced (larvae removed), the concentration of Vc in orange juice was 18.65 µg/mL, which was 9.16 µg/mL lower than that measured in healthy orange juice. In addition, the total phenol content decreased by 46.519 mg Gallic Acid Equivalents (GAE)/g to 9.748 mg GAE/g. Furthermore, the free-radical scavenging activity decreased from 22.297% to 5.393%. Correlation analysis indicated significant correlations between Vc concentration, total phenol content, and antioxidant activity of orange juice after B. dorsalis infestation. The decrease in Vc concentration, total phenol content and free-radical scavenging activity indicated that B. dorsalis changed the quality of orange juice by affecting the antioxidant activity of the juice after the oranges were infested.

Resumo Entre frutas e produtos de frutas, as laranjas e suco de laranja são os mais consumidos em todo o mundo. No entanto, os efeitos da infestação de laranjas sobre a qualidade do suco de laranja ainda não são conhecidos. Para avaliar o efeito da mosca-das-frutas oriental, Bactrocera dorsalis, sobre a atividade antioxidante do suco de laranja, foram medidas as mudanças na concentração de vitamina C (Vc), no teor de fenol total e na atividade antioxidante do suco de laranja após a introdução de ovos da mosca-das-frutas oriental. Dez dias após a introdução dos ovos (larvas removidas), a concentração de Vc no suco de laranja foi de 18,65 µg/mL, que foi 9,16 µg/mL menor do que a medida em suco de laranja saudável. Além disso, o teor total de fenol diminuiu de 46,519 mg em equivalente de ácido gípico (GAE)/g para 9,748 mg de GAE/g. Ademais, a atividade de eliminação de radicais livres diminuiu de 22,297% para 5,393%. A análise de correlação indicou correlações significativas dentre a concentração de Vc, o conteúdo total de fenol e a atividade antioxidante do suco de laranja após a infestação por B. dorsalis. A diminuição na concentração de Vc, o conteúdo total de fenol e a atividade sequestradora de radicais livres indicaram que B. dorsalis alterou a qualidade do suco de laranja, afetando a atividade antioxidante do suco após a infestação das laranjas.

Expression of anoctamin 1 is associated with advanced tumor stage in patients with non-small cell lung cancer and predicts recurrence after surgery.

PURPOSE: Anoctamin 1 (ANO1), a recently identified calcium-activated chloride channel, has been found to have a critical role in tumorigenesis and tumor progression in several types of cancer. However, its role in non-small cell lung cancer (NSCLC) remains to be elucidated. In this study, we evaluated the utility of ANO1 as a prognostic marker. PATIENTS AND METHODS: ANO1 expression was detected in tumor tissues and paraneoplastic tissues of I-IV stage NSCLC patients who received surgical treatment by using immunohistochemical and quantitative RT-PCR analyses. Epidermal growth factor receptor (EGFR) was investigated using immunohistochemistry. Then the TNM stage of the tumor samples was assessed and patients were followed up for developing recurrence. RESULTS: ANO1 expression was significantly increased in NSCLC tumor tissues compared to the paraneoplastic tissues at both RNA and protein level. In addition, ANO1 overexpression was correlated with the high expression of EGFR and led to an advanced tumor stage. And also high ANO1 expression was significantly correlated with high recurrence rate at 1-year follow-up. CONCLUSIONS: ANO1 overexpression associated with the high expression of EGFR can be a predictive marker of recurrence after surgery in NSCLC patients.

Effect of oviposition by Bactrocera dorsalis on the antioxidant activity of orange juice

Abstract Among fruits and fruit products, oranges and orange juice are the most widely consumed worldwide. However, the effects of pest infestation of oranges on the quality of orange juice are not yet known. To evaluate the effect of the oriental fruit fly Bactrocera dorsalis on the antioxidant activity of orange juice, we measured changes in the vitamin C (Vc) concentration, total phenol content, and antioxidant activity of orange juice after the introduction of fruit fly eggs. Ten days after the eggs were introduced (larvae removed), the concentration of Vc in orange juice was 18.65 µg/mL, which was 9.16 µg/mL lower than that measured in healthy orange juice. In addition, the total phenol content decreased by 46.519 mg Gallic Acid Equivalents (GAE)/g to 9.748 mg GAE/g. Furthermore, the free-radical scavenging activity decreased from 22.297% to 5.393%. Correlation analysis indicated significant correlations between Vc concentration, total phenol content, and antioxidant activity of orange juice after B. dorsalis infestation. The decrease in Vc concentration, total phenol content and free-radical scavenging activity indicated that B. dorsalis changed the quality of orange juice by affecting the antioxidant activity of the juice after the oranges were infested.

Resumo Entre frutas e produtos de frutas, as laranjas e suco de laranja são os mais consumidos em todo o mundo. No entanto, os efeitos da infestação de laranjas sobre a qualidade do suco de laranja ainda não são conhecidos. Para avaliar o efeito da mosca-das-frutas oriental, Bactrocera dorsalis, sobre a atividade antioxidante do suco de laranja, foram medidas as mudanças na concentração de vitamina C (Vc), no teor de fenol total e na atividade antioxidante do suco de laranja após a introdução de ovos da mosca-das-frutas oriental. Dez dias após a introdução dos ovos (larvas removidas), a concentração de Vc no suco de laranja foi de 18,65 µg/mL, que foi 9,16 µg/mL menor do que a medida em suco de laranja saudável. Além disso, o teor total de fenol diminuiu de 46,519 mg em equivalente de ácido gípico (GAE)/g para 9,748 mg de GAE/g. Ademais, a atividade de eliminação de radicais livres diminuiu de 22,297% para 5,393%. A análise de correlação indicou correlações significativas dentre a concentração de Vc, o conteúdo total de fenol e a atividade antioxidante do suco de laranja após a infestação por B. dorsalis. A diminuição na concentração de Vc, o conteúdo total de fenol e a atividade sequestradora de radicais livres indicaram que B. dorsalis alterou a qualidade do suco de laranja, afetando a atividade antioxidante do suco após a infestação das laranjas.

Cytotoxicity and DNA damage in mouse macrophages exposed to silica nanoparticles.

Silica (SiO2) nanoparticles are being progressively applied in various applications, including cosmetics, food technology, and medical diagnostics. Although crystalline SiO2 is a known carcinogen, the carcinogenicity of SiO2 nanoparticles remains unclear. Here, we assessed the cytotoxic effects and DNA injury induced by exposure to various dosages of SiO2 nanoparticles at 0-2400 mg/mL (0-3200 mg/mL microscale SiO2 as positive control) for 24 h using RAW264.7 cells, followed by methyl tetrazolium (MTT) assay. Cells were also treated by 31.25, 125, and 500 mg/mL SiO2 nanoparticles (500 mg/mL microscale SiO2 as positive control) for 24 h and examined by single cell gel electrophoresis assay (SCEG) and flow cytometry. Outstanding dose-related decline in cell viability was observed with enhancing dosages of SiO2 nanoparticles by MTT assay. The inhibitory concentration 50% of SiO2 nanoparticles and microscale SiO2 was 16690 and 5080 mg/mL, respectively. The comet rate (comet%), length of tail, the percentage in DNA tail (TDNA%) and olive tail moment (OTM) induced by SiO2 nanoparticles were significantly increased in comparison with control and microscale SiO2 at 500 mg/mL. 500 mg/mL SiO2 nanoparticles and microscale SiO2 caused a significant increase in apoptosis rate, decreased proliferation index and increased cell proportions in G0/G1 phases by contrast to the negative control (P < 0.05). This indicates that SiO2 nanoparticles are more cytotoxic than microscale SiO2 particles; they induce DNA injury, increase apoptosis, and decrease the proliferation index in RAW264.7 cells. DNA injury and apoptosis may be involved in reducing cell proliferation.

Effect of polychlorinated biphenyls on oxidation stress in the liver of juvenile GIFT, Oreochromis niloticus.

The present study clearly showed that chronic exposure to polychlorinated biphenyls (PCBs) at environmentally relevant concentrations can damage juvenile tilapia livers by modulating antioxidant enzyme activities and gene transcription, which affects toxic bioaccumulation and histological congestion. The results suggest that PCBs caused a decrease in the activity of some hepatic antioxidative and biotransformation enzymes (SOD, CAT, GST, T-GSH, and MDA) in tilapia at 7 days, as well as transcriptional changes (sod, cat, and gst). Except for some antioxidant parameters (T-GSH, GSH/GSSG, T-AOC, and MDA), significant declines and increases occurred at 14 and 21 days, respectively. Most of the antioxidant enzymatic signatures and genotoxicity significantly increased at 14 and 21 days. This study presented evidence that PCBs could result in hepatic toxicity through oxidative stress in the early growth stages of tilapia, and we speculated that oxidative stress plays an important role in embryonic developmental toxicity induced by PCBs.

Effect of polychlorinated biphenyls on osmoregulatory response and apoptosis in GIFT tilapia, Oreochromis niloticus.

In the present study, GIFT tilapia Oreochromis niloticus were exposed to polychlorinated biphenyls (PCBs) for 7, 14, and 21 days. Over the duration of the exposure, genotoxicity and the activity of Na+/K+-ATPase (NKA) and Ca+/Mg+-ATPase (CMA) were measured in the gill, kidney, and intestine, to evaluate changes in osmoregulatory response in O. niloticus. Our results showed significant decreases in organic NKA (except in gill tissues after 0.5 mg/L PCB-exposure) and CMA activity. The results of the genotoxicity assay showed significant increases in atp1a1a, nkcc2 (only in gill tissue), and fxyd7 (except after 21 days of 5 mg/L PCB exposure). We found significant increases in caspase proteins in the liver in the 5-mg/L PCB exposure group, and the transcripts showed dose-dependent increases between treatment groups over the exposure duration. This study presents evidence that chronic exposure to PCB could result in organic osmoregulatory response and hepatic apoptosis in GIFT tilapia.

Correlation between polymorphisms in the visfatin gene and its expression in the serum and coronary artery calcification.

We investigated the association between serum visfatin levels and single nucleotide polymorphisms (SNPs; rs61330082, rs2058539) in the visfatin gene and coronary artery calcification (CAC) in patients from Wenzhou, China. CAC patients (N = 206) were divided into two groups: mild CAC (MCAC) and moderate and severe CAC (MSCAC). Volunteers without CAC (N = 70) were included in the control group. The serum visfatin level was analyzed by enzyme-linked immunosorbent assay. SNPs (rs61330082, rs2058539) in the visfatin gene were analyzed by polymerase chain reaction-restriction fragment length polymorphism. Clinical data, serum visfatin levels, and genotype and allele frequencies of rs61330082 and rs2058539 were compared among the three groups. MSCAC patients expressed significantly higher serum visfatin levels (30.58 ± 6.12 ng/mL) than individuals in the MCAC (29.03 ± 1.87 ng/mL) and control (24.45 ± 5.44 ng/mL) groups (P < 0.05). The genotype distributions and frequencies of rs61330082 differed significantly among the groups (P < 0.05), while those of rs2058539 did not. The serum visfatin level was positively correlated with the body mass index (BMI), high-density lipoprotein cholesterol (HDL-C), and insulin resistance index (IRI), and negatively correlated with the triglyceride (TG) levels (P < 0.05) of patients. Serum visfatin is associated with the development of CAC. The T allele of the rs61330082 SNP in the visfatin gene had a cardioprotective effect on patients with CAC; the SNP at rs2058539 was not significantly associated with CAC. The BMI, HDL-C, IRI, and TG levels influenced the development of CAC.

Prevalence of and factors associated with frailty and disability in older adults from China, Ghana, India, Mexico, Russia and South Africa.

BACKGROUND: The severe burden imposed by frailty and disability in old age is a major challenge for healthcare systems in low- and middle-income countries alike. The current study aimed to provide estimates of the prevalence of frailty and disability in older adult populations and to examine their relationship with socioeconomic factors in six countries. METHODS: Focusing on adults aged 50+ years, a frailty index was constructed as the proportion of deficits in 40 variables, and disability was assessed using the World Health Organization Disability Assessment Schedule (WHODAS 2.0), as part of the Study on global AGEing and adult health (SAGE) Wave 1 in China, Ghana, India, Mexico, Russia and South Africa. RESULTS: This study included a total of 34,123 respondents. China had the lowest percentages of older adults with frailty (13.1%) and with disability (69.6%), whereas India had the highest percentages (55.5% and 93.3%, respectively). Both frailty and disability increased with age for all countries, and were more frequent in women, although the sex gap varied across countries. Lower levels of both frailty and disability were observed at higher levels of education and wealth. Both education and income were protective factors for frailty and disability in China, India and Russia, whereas only income was protective in Mexico, and only education in South Africa. CONCLUSIONS: Age-related frailty and disability are increasing concerns for older adult populations in low- and middle-income countries. The results indicate that lower levels of frailty and disability can be achieved for older people, and the study highlights the need for targeted preventive approaches and support programs.

Fine mapping and characterization of the or gene in Chinese cabbage (Brassica rapa L. ssp pekinensis).

Orange inner leaves/heads is a qualitative trait in Chinese cabbage that is controlled by a single recessive gene. Orange head Chinese cabbage contain more carotenoids than its white head counterpart; hence, this trait is of interest to both researchers and consumers. In this study, we selected the orange head Chinese cabbage line 07A163 and the white head Chinese cabbage line Chiifu as test materials. We localized the target gene controlling the orange head trait to the A09 linkage group, with a physical distance of approximately 19.9 kb between the two markers, syau26 and syau28. This region contains six candidate genes, including Bra031539, which was predicted to encode CRTISO, a carotenoid isomerase specifically required for carotenoid biosynthesis. A comparison of the nucleic acid sequences of the two test materials revealed 88 and 7-bp deletions and 88 SNPs in the promoter region of Bra031539 in line 07A163, along with a 6-bp deletion in the first exon and early termination at the 3' end of this gene. BLAST analysis revealed that 22 amino acids were altered and 17 amino acids were lost in Bra031539 in the orange head line 07A163. We developed the BrPro1 molecular marker in the promoter region of Bra031539 that can be used for early identification of orange head materials, thereby accelerating the breeding process of orange head Chinese cabbage.

Evaluation of the semen swim-up method for bovine sperm RNA extraction.

Isolation of high-quality RNA is important for assessing sperm gene expression, and semen purification methods may affect the integrity of the isolated RNA. This study evaluated the effectiveness of the sperm swim-up method for seminal RNA isolation. Frozen semen samples in straws from three bulls of proven fertility were purified by the swim-up method. RNA extraction was carried out using the E.Z.N.A.(TM) Total RNA kit II, with non-swim-up sperm as a control. Total sperm RNA was analyzed by UV spectrophotometry, reverse transcription polymerase chain reaction (RT-PCR), and agarose gel electrophoresis, and expression of the sex-determining region on the Y chromosome (SRY), leptin (LEP), and ribosomal protein subunit 23 (RPS23) genes, were determined. 18S RNA was used as a positive control. Fewer somatic cells were found in sperm swim-up samples than in the non-swim-up counterparts (0 x 10(3) vs 17.33 ± 2.52 x 10(3) sperm, P < 0.05). In addition, high-quality RNA was obtained in about 2 h, with no significant difference between groups. Interestingly, the yields of RNA fragments containing ≥200 nucleotides were significantly reduced in sperm swim-up samples (0.92 ± 0.41 x 10(7) sperm) compared with the non-swim-up samples (1.36 ± 0.33 x 10(7) sperm, P < 0.05). After RT-PCR, clear bands representing SRY, LEP, and RPS23 in sperm cDNA were observed on agarose gel electrophoresis. Finally, no bands corresponding to 18S RNA were found in RNA samples from the sperm swim-up group. Our findings suggest that small amounts of sperm RNA can be efficiently extracted from frozen straw semen samples using the swim-up technique.

Molecular identification and genetic variation of varieties of Styphnolobium japonicum (Fabaceae) using SRAP markers.

Thirty-four Styphnolobium japonicum varieties were analyzed using sequence-related amplified polymorphism (SRAP) markers, to investigate genetic variation and test the effectiveness of SRAP markers in DNA fingerprint establishment. Twelve primer pairs were selected from 120 primer combinations for their reproducibility and high polymorphism. We found a total of 430 amplified fragments, of which 415 fragments were considered polymorphic with an average of 34.58 polymorphic fragments for each primer combination. The percentage of polymorphic fragments was 96.60%, and four primer pairs showed 100% polymorphism. Moreover, simple matched coefficients ranged between 0.68 and 0.89, with an average of 0.785, indicating that the genetic variation among varieties was relatively low. This could be because of the narrow genetic basis of the selected breeding material. Based on the similarity coefficient value of 0.76, the varieties were divided into four major groups. In addition, abundant and clear SRAP fingerprints were obtained and could be used to establish DNA fingerprints. In the DNA fingerprints, each variety had its unique pattern that could be easily distinguished from others. The results demonstrated that 34 varieties of S. japonicum had a relatively narrow genetic variation. Hence, a broadening of the genetic basis of breeding material is necessary. We conclude that establishment of DNA fingerprint is feasible by means of SRAP markers.

Large-scale tissue-specific and temporal gene expression profiles in Pengze crucian carp.

In the present study, the tissue-specific and temporal gene expression profiles of four catalogues of gonadal development-related genes (sex differentiation-related, steroid receptor, steroidogenic, and structural genes) were detected in nine tissues and during 11 successive developmental stages in the Pengze crucian carp (Pcc) (a triploid mono-female gynogenic fish). The results showed that these target genes exhibited overlapping distributions in various tissues, with the exception of Pcc-vasa and Pcc-cyp17a1. Gene expression profiling of the developmental stages showed that all of the target genes simultaneously reached peak expression levels at 40 and 48 days post hatching (dph). Both 40 and 48 dph appeared to be two key time points associated with the process of Pcc gonadal development. These data will provide a clear understanding of gene expression patterns associated with the gonadal development-related genes of this gynogenic teleost.

Karyotype studies on Lycoris radiata populations from China.

Lycoris radiata is an important medicinal and ornamental plant of China. In the present study, somatic chromosome counts and karyotype analyses, which are important aspects of plant phylogeny and evolution, were performed in 466 individuals from 25 L. radiata populations by root tip squash method. Chromosome counts revealed that 10 populations were diploid (2n = 2x = 22) and 15 were triploid (2n = 3x = 33). Except for one diploid population containing some triploid plants, the remaining 24 populations showed a single cytotype. Karyotype analysis showed that the karyotypes of L. radiata varied in different populations and even within the same population. However, based on the Stebbins' system, the karyotype of all the populations could be classified in 4A classes. The cluster analysis and ordination methods demonstrated that the L. radiata populations grouped in two major clusters. Previous research has shown that the triploid strain of L. radiata is a genetically identical species. However, the cluster analysis revealed that the triploid strains clustered in two groups instead of one, which indicates that these strains may not be identical species, genetically. This study is expected to improve the understanding of the genetic diversity in L. radiata and provide a basis for future studies on species differentiation, speciation, and taxonomy.

Development of microsatellite loci for Taxus wallichiana var. wallichiana (Taxaceae) and cross-amplification in Taxaceae.

Nine polymorphic microsatellite loci were isolated and characterized in Taxus wallichiana var. wallichiana, an endangered species in China. The number of alleles per locus ranged from 2 to 20. Observed and expected heterozygosities varied from 0.0260 to 0.5325 and 0.3603 to 0.9231, respectively. Positive cross-amplification of the 9 loci was observed in 2 other varieties of T. wallichiana and 4 other Taxaceae species. These loci will be of value for studying population genetic structures and for genetic resource conservation in T. wallichiana and other Taxus species.

Modification research on in wall of capillary copper tube with Norland optical adhesive 68 in a double stereo PCR microfluidic chip.

In this study, a Norland optical adhesive 68 (NOA68) film, approximately 2.2 µm thick, was manufactured using ultraviolet solidified NOA68 in apparatus manufacturing film on the inwall of a capillary copper pipe, developed in our laboratory. The roughness of the inwall of capillary copper pipe was improved from Ra = 0.766 to 0.204 µm and the contact angle was improved from approximately 96° to 55°, increasing hydrophilicity. Polymerase chain reaction experiments indicated that the ratio of work pressure in the microfluidic chip before modification to that after modification was 2.71/1, indicating that the extension efficiency increased. Our results provide a basis for the construction of a microform chip based on function integration.