A method was developed for determination of dilauryl thiodipropionate(DLTDP)in fried foods by coupling solid-phase extraction(SPE)pretreatment with reverse-phase liquid chromatography-tandem mass spectrometry(RPLC-MS/MS)detection.Samples were extracted with n-hexane as the solvent,purified using a neutral alumina SPE cartridge,and finally analyzed by RPLC-MS/MS.Quantitative analysis was performed using matrix-matched calibration curves combined with an external standard method under optimal experimental conditions.The results showed that DLTDP exhibited good linearity in the range of 2.0-50.0 μg/L,with a correlation coefficient(R2)≥0.999.The limit of detection(LOD)and the limit of quantification(LOQ)of the method were 0.15 mg/kg and 0.5 mg/kg,respectively.The mean recoveries at three fortification levels(0.5,1.0,and 200 mg/kg)in different samples ranged from 84.8%to 96.8%,with the relative standard deviations(RSDs)all less than 8.0%.The developed method was highly sensitive,accurate and reliable,and easy to operate,making it well suited for the routine quantitative analysis of DLTDP in fried foods.

A 31P-quantitative nuclear magnetic resonance(qNMR)method was established for simultaneous determination of sodium α-glycerophosphate,sodium β-glycerophosphate and phosphoric acid in concentrated divitamins and sodium phosphate syrup.The qNMR experimental conditions were optimized,including hexamethylphosphoramide as internal standard,20%deuterium oxide solution as solvent,zgig pulse sequence,delay time of 30 s,and scan number of 64.The 31P-NMR peaks atδ29.80 of hexamethylphosphoramide,δ0.69 of sodiumα-glycerophosphate,δ0.17 of sodiumβ-glycerophosphate andδ0.03 of phosphoric acid were chosen as the quantitative peaks(pH of the test solution was around 4.8).Method validation was performed in terms of precision(Relative standard deviation less than 0.6%),linearity(Correlative coefficient greater than 0.999),limit of detection(23.58 μg/mL for sodium glycerophosphate and 11.61 μg/mL for phosphoric acid)and limit of quantitation(78.60 μg/mL for sodium glycerophosphate and 38.70 μg/mL for phosphoric acid).The recoveries were 99.8%?103.2%,and relative standard deviations were 0.41%?1.98%.The results showed that the reliability of 31P-qNMR method were suitable for its intended use.Seven batches of concentrated divitamins and sodium phosphate syrups were tested by the established method,of which the total phosphorus content was consistent with that of colorimetry method,but the content of sodium glycerophosphate(Sum ofαtype andβtype)was relatively low,about 82%of the labeled amount.The content of phosphoric acid was high.This method simplified sample pretreatment and had high specificity,and was more suitable for determination and quality control of concentrated divitamins and sodium phosphate syrup.

OBJECTIVE: This study was aimed at investigating the carrier rate of, and molecular variation in, α- and β-globin gene mutations in Hunan Province. METHODS: We recruited 25,946 individuals attending premarital screening from 42 districts and counties in all 14 cities of Hunan Province. Hematological screening was performed, and molecular parameters were assessed. RESULTS: The overall carrier rate of thalassemia was 7.1%, including 4.83% for α-thalassemia, 2.15% for β-thalassemia, and 0.12% for both α- and β-thalassemia. The highest carrier rate of thalassemia was in Yongzhou (14.57%). The most abundant genotype of α-thalassemia and β-thalassemia was -α ^3.7/αα (50.23%) and β ^IVS-II-654/β ^N (28.23%), respectively. Four α-globin mutations [CD108 (ACC>AAC), CAP +29 (G>C), Hb Agrinio and Hb Cervantes] and six β-globin mutations [CAP +8 (C>T), IVS-II-848 (C>T), -56 (G>C), beta nt-77 (G>C), codon 20/21 (-TGGA) and Hb Knossos] had not previously been identified in China. Furthermore, this study provides the first report of the carrier rates of abnormal hemoglobin variants and α-globin triplication in Hunan Province, which were 0.49% and 1.99%, respectively. CONCLUSION Our study demonstrates the high complexity and diversity of thalassemia gene mutations in the Hunan population. The results should facilitate genetic counselling and the prevention of severe thalassemia in this region.
Humans ; beta-Thalassemia/genetics* ; alpha-Thalassemia/genetics* ; Hemoglobinopathies/genetics* ; China/epidemiology* ; High-Throughput Nucleotide Sequencing

OBJECTIVE: To investigate the expression of MC1R in esophageal squamous cell carcinoma and its correlation with the clinicopathological parameters. METHODS: We analyzed the expression of MC1R in esophageal cancer based on data from TCGA databse and examined its expression levels using RT-PCR and Western blotting in a human esophageal epithelial cell line BAr-T, human esophageal squamous cell carcinoma cell lines ECA109, KYSE30, KYSE150, KYSE510, TE-1, TE-13, and EC9706, a human gastric cancer cell line SGC7901 and 19 pairs of esophageal squamous cell carcinoma tissues and adjacent tissues.Immunohistochemistry was used to detect MC1R expression levels in 32 pairs of paraffin-embedded sections of esophageal squamous cell carcinoma and adjacent tissues, and the correlation of MC1R expression and the patients'clinicopathological characteristics was analyzed. RESULTS: Bioinformatics analysis showed that MC1R was significantly overexpressed in esophageal cancer tissues (P < 0.05).MC1R expression was also increased in 5 esophageal squamous cell carcinoma cell lines ECA109, KYSE30, KYSE510, TE-13, EC9706 and the gastric cancer cell line SGC7901 as compared with that in esophageal epithelial cells (P < 0.05).Immunohistochemistry revealed significantly increased MC1R expression in esophageal squamous cell carcinoma tissue sections in comparison with the adjacent tissue sections (P < 0.05).In patients with esophageal squamous cell carcinoma, a high MC1R expression was detected mainly in those with an old age, positive for middle-thoracic involvement, and with moderately differentiated tumor cells, and showed a correlation with T stage of tumor (P < 0.05), but not with the other clinicopathological parameters such as gender, age, degree of cell differentiation, primary tumor site, or TNM stage (P>0.05). CONCLUSION MC1R is highly expressed in esophageal squamous cell carcinoma and may serve as a molecular biomarker to assist in the diagnosis of esophageal squamous cell carcinoma.
Humans ; Esophageal Squamous Cell Carcinoma ; Esophageal Neoplasms/metabolism* ; Stomach Neoplasms ; Cell Line, Tumor ; Immunohistochemistry ; Cell Proliferation ; Gene Expression Regulation, Neoplastic

BACKGROUND: Recent studies suggest that a healthy diet helps to prevent the development of Alzheimer disease (AD). This study aimed to investigate whether spicy food consumption is associated with cognition and cerebrospinal fluid (CSF) biomarkers of AD in the Chinese population. METHODS: We enrolled 55 AD patients and 55 age- and gender-matched cognitively normal (CN) subjects in a case-control study, as well as a cohort of 131 participants without subjective cognitive decline (non-AD) in a cross-sectional study. Spicy food consumption was assessed using the Food Frequency Questionnaire (FFQ). Associations of FFQ scores with cognition and CSF biomarkers of AD were analyzed. RESULTS: In the case-control study, spicy food consumption was lower in AD patients than that in CNs (4.0 [4.0-8.0] vs. 8.0 [4.5-10.0], P < 0.001); FFQ scores were positively associated with Mini-Mental Status Examination scores in the total sample (r = 0.218, P = 0.014). In the cross-sectional study, the association between spicy food consumption and cognition levels was verified in non-AD subjects (r = 0.264, P = 0.0023). Moreover, higher FFQ scores were significantly associated with higher β-Amyloid (1-42) (Aβ42) levels and lower phospho-tau/Aβ42 and total tau/Aβ42 ratios in the CSF of non-AD subjects (P < 0.05). CONCLUSION Spicy food consumption is closely related to higher cognition levels and reversed AD biomarkers in the CSF, suggesting that a capsaicin-rich diet might have the potential to modify the cognitive status and cerebral pathologies associated with AD.
Alzheimer Disease ; Amyloid beta-Peptides ; Biomarkers ; Case-Control Studies ; Cognition ; Cross-Sectional Studies ; Humans ; Peptide Fragments ; tau Proteins

AlM:To investigate the effects of pirfenidone ( PFD) on the proliferation and transfomring growth factor-β1 ( TGF-β1 ) expression in vitro culture rat corneal stromal cells.METHODS: Corneal stromal cells from 8 to 10wk SD rats were isolated, cultured and treated with different concentrations of PFD 0mg/mL (control group), 0. 15mg/mL (experimental group▏), 0. 3mg/mL (experimental group‖), 1mg/mL (experimental group Ⅲ) for 48h. CCK-8 assay was performed to assess cell proliferation, while immunocytochemistry and Western Blot were used to detect the expression of ki-67 and TGF-β1 expression, respectively. RESULTS: Compared with control group, PFD significantly inhibited the proliferation in a dose -dependent manner ( all P < 0. 05 ), so was protein expression of ki-67. PFD significantly down-regulated the expression of TGF-β1 in a dose-dependent manner (P<0. 05).CONCLUSlON: Pirfenidone can significantly inhibit the proliferation of rat corneal stromal cell by down regulating TGF-β1 expression, therefore, it has potential prospect in lightening the corneal wound healing reaction.

OBJECTIVETo investigate the safety of assisted reproductive technology (ART) with donated sperm from the sperm bank and the differences in the pregnancy outcomes of different means of promoting pregnancy.

METHODSWe analyzed and compared the feedback data on promoting pregnancy with donated sperm from the sperm bank by artificial insemination by donor (AID), in vitro fertilization (IVF), and intracytoplasm sperm injection (ICSI).

RESULTSTotally, 13 723 tubes of sperm specimens were used for ART. The number of specimens used differed in different clinical reproductive centers, some using 1 tube and others using 2 tubes per cycle. The 13 723 tubes were used for a total of 7 743 cycles. Among the 7 123 cycles of AID, there were 1 415 clinical pregnancies (19.87%), 1 221 normal births (86.29%), 169 abortions (11.94%), 6 cases of birth defects (0.43%), 19 ectopic pregnancies (1.34%), and 0 sexually transmitted infection. Among the 571 cycles of IVF, there were 367 clinical pregnancies (64.27%), 330 normal births (89.92%), 35 abortions (9.54%), 0 birth defect, 2 ectopic pregnancies (0.54%), and 0 sexually transmitted infection. Among the 49 cycles of ICSI, there were 28 clinical pregnancies (57.14%), 25 normal births (89.29%), 3 abortions (10.71%), 0 birth defect, 0 ectopic pregnancy, and 0 sexually transmitted infection. There were statistically significant differences in the rate of clinical pregnancy among AID, IVF and ICSI (P < 0.05), but not between IVF and ICSI (P > 0.05), nor were there any significant differences in the rates of abortion, birth defects and ectopic pregnancy among AID, IVF and ICSI (P > 0.05).

CONCLUSIONNone of the recipients of the donated sperm from the sperm bank was infected with sexually transmitted diseases. AID, IVF and ICSI showed no significant differences from natural conception in the rates of abortion, birth defects and ectopic pregnancy. ART with donated sperm from the sperm bank is safe. IVF and ICSI are associated with a higher rate of pregnancy than AID, though the latter costs less than the former two.

Female ; Fertilization in Vitro ; Humans ; Male ; Pregnancy ; Pregnancy Outcome ; Pregnancy Rate ; Sperm Banks ; Sperm Injections, Intracytoplasmic ; Spermatozoa

OBJECTIVETo investigate the effect of high mobility group box-1 (high mobility group box B 1, HMGB1) on the invasive and metastatic abilities of gastric cancer cell line MGC-803 and analyze the possible mechanisms.

METHODSHMGB1 gene targeting siRNA was designed and synthesized, and HMGB1 siRNA oligonucleotides were transfected into the MGC-803 cells with Lipofectamine 2000. The invasive and migratory abilities were detected by transwell assay and scratch assay. The Matrigel matrix glue adhesive ability of MGC-803 cells was evaluated by MTT assay. NF-κB activity was detected by electrophoretic mobility shift assay. The mRNA and protein levels of HMGB1 and MMP-9 were determined by RT-PCR and Western blot, respectively.

RESULTSThe siRNA down-regulated the levels of HMGB1 mRNA and protein. Compared with that of the control group, the number of invasive (142.7 ± 3.4 /view vs. 303.5 ± 4.3/view) and migratory (293.7 ± 4.4/view vs. 445.5 ± 5.6/view) cells was significantly increased (P < 0.05) and the adhesive ability of MGC-803 cells to Matrigel was significantly elevated (33.4 ± 0.03% vs. 57.4 ± 4.2%, P < 0.05). In addition, silencing of HMGB1 gene significantly inhibited the activity of NF-κB and the relative expression folds of mRNA (0.2 ± 0.1 vs. 1.4 ± 0.4, P < 0.05)and protein (0.4 ± 0.1 vs. 2.3 ± 0.7, P < 0.05) of MMP-9.

CONCLUSIONSilencing of HMGB1 can effectively inhibit the invasion and migration of gastric cancer cells and this effect of HMGB1 may be partly due to its regulation of NF-κB and MMP-9 expressions.

Cell Adhesion ; Cell Line, Tumor ; Cell Movement ; Down-Regulation ; Gene Expression Regulation, Neoplastic ; HMGB1 Protein ; genetics ; metabolism ; Humans ; Matrix Metalloproteinase 9 ; genetics ; metabolism ; NF-kappa B ; genetics ; metabolism ; Neoplasm Invasiveness ; Neoplasm Metastasis ; RNA, Messenger ; metabolism ; RNA, Small Interfering ; genetics ; Stomach Neoplasms ; metabolism ; pathology ; Transfection

The application of extralevator abdomino-perineal excision (ELAPE) and total mesorectal excision has improved the prognosis of rectal cancer. However, compared with anterior resection for rectal cancer, the circumferential resection margin (CRM) positive rate and intraoperative perforation (IOP) rate are still high. The ELAPE can reduce the CRM positive rate and IOP rate, therefore reduce postoperative local recurrence rate and increase the survival rate of patients. The disadvantage of its trauma, longer operative time, and higher perineum complication in ELAPE is controversial. This review mainly discusses the key points of operative procedure, advantages and disadvantages, research status and development prospects of ELAPE.
Digestive System Surgical Procedures ; methods ; Humans ; Mesentery ; surgery ; Perineum ; surgery ; Prognosis ; Rectal Neoplasms ; surgery ; Rectum ; surgery

Objective To observe protective effects on rat serum cardiac enzymes and the antioxidant capacity of selenium and vitamin E.Methods According to body weight and 2 × 2 factorial design,eighty male Wistas rats were randomly divided into four groups:low selenium and low vitamin E group(feed containing 23.42％ of the low selenium yeast,excluding vitamin E),low selenium and adequate vitamin E group (feed containing 23.42％ of the low selenium yeast and vitamin E 160 mg/kg),adequate selenium and low vitamin E group(feed containing 46.84％ of the low selenium yeast and sodium seleni 0.25 mg/L in water,excluding vitamin E),adequate selenium and adequate vitamin E group(feed containing 46.84％ of the low selenium yeast,vitamin E 160 mg/kg and sodium selenite 0.25 mg/L in water),20 rats every group.Rats were feed with synthetic feed,and given intraperitoneal anesthesia after 26 weeks of feeding.Blood was collected to observe the impact of selenium and vitamin E on rat cardiac enzymes and myocardial antioxidant capacity and their interactions.Serum creatine kinase (CK) was measured using the continuous monitoring method,creatine kinase isozymes (CK-MB) and lactate dehydrogenase(LDH ) using the immune suppression method,the whole blood GSH-Px assay using the dithiobis nitrohenzoic acid(DTNB) method,serum superoxide dismutase(SOD) using the xanthine oxidase method,total antioxidant capacity (T-AOC) using the complex colorimetry method,the content of propylene glycol (MDA) using the thiobarbituric acid colorimetric method,and reactive oxygen species(ROS) using the colorimetric method.Results Group differences of serum CK,CK-MB,LDH,whole blood GSH-Px activity,serum T-AOC vitality,MDA and ROS content were statistically significant(F=9.797,17.041,48.399,3.744,224.900,49.384,5.045,all P＜ 0.05).Compared with the two low selenium groups and one adequate selenium group,the vitalities of CK,CK-MB,LDH and the contents of MDA[(1577.75 ± 451.87),(1239.15 ± 344.99),(884.25 ± 133.84)U/L,(5.688 ±1.169) × 103 nmol/L; (1474.21 ± 398.38),(1014.84 ± 215.40),(523.00 ± 98.05)U/L,(4.035 ± 0.487 ) × 103 nmol/L and (1180.10 ± 245.51),(948.75 ± 173.68),(676.70 ± 193.63)U/L,(3.406 ± 0.146) × 103 nmol/L]increased significantly in adequate selenium and adequate vitamin E group[( 1056.80 ± 250.98),(721.70 ±129.98),(404.65 ± 72.49)U/L,(3.010 ± 1.270) × 103 nmol/L,all P ＜ 0.05) ].The activity of GSH-Px was obviously increased in the two adequate selenium groups[ (96.611 ± 8.238) × 103,(103.024 ± 8.217) × 103 U/L,all P ＜ 0.05],compared with the two low selenium groups[ (60.356 ± 8.179) × 103,(63.117 ± 8.281) × 103 U/L].Selenium affected the activities of CK,CK-MB and LDH(F =27.09,31.58,29.66,all P＜ 0.01 ),and vitamin E affected the activities of CK-MB and LDH(F=18.9,11.2.all P＜ 0.01 ),but both selenium and vitamin E had no interactions on the activities of CK,CK-MB and LDH (F=0.02,0.001,2.22,all P＞0.05).Selenium affected the activity of GSH-Px and the content of MDA(F=6.74,95.68,all P＜ 0.05),vitamin E affected the activity of T-AOC,the contents of MDA and ROS(F=6.42,36.73,8.43,all P＜0.05),but selenium and vitamin E had interactions only on the content of MDA(F =13.82,P＜ 0.05).Conclusions Long-term selenium or vitamin E deficiency,can reduce the body's antioxidant capacity,leading to the occurrence of myocardial injury.Selenium and vitamin E can improve the body's oxidation capacity,playing a role in myocardial protection.