OBJECTIVE To determine the contents of 15 components in Aurantii Fructus Immaturus from different origins （Citrus aurantium， C. junos， C. aurantium Linn.， C. sinensis Osb.， C. sinensis）， screen differential components， and provide references for the quality evaluation of Aurantii Fructus Immaturus. METHODS HPLC method was adopted to determine the contents of synephrine， N-methyltyramine， 5，7-dihydroxychromone-7-neohesperidoside， neoponcirin， narirutin， naringin， hesperidin， neohesperidin， naringenin， hesperetin， sinensetin， nobiletin， tangeretin， 5-demethylnobiletin， and auraptene in 46 batches of Aurantii Fructus Immaturus from different origins. The determination was performed on Waters Symmetry C18 column with mobile phase consisted of acetonitrile-0.1% formic acid （gradient elution） at the flow rate of 1.0 mL/min； column temperature was set at 40 ℃ ， detection wavelength was 284 nm， and sample injection volume was 5 μL. The differences between different origins of Aurantii Fructus Immaturus were analyzed by cluster analysis， principal component analysis （PCA） and orthogonal partial least squares-discriminant analysis （OPLS-DA）， and differential components were screened. RESULTS The linear relationships of the aforementioned 15 components were all good within the tested mass concentration ranges （all r＞0.999 0）. The RSDs for precision， stability （24 h）， and repeatability tests were all less than 2.00%. The average recovery rate ranged from 91.1% to 103.9% （all RSDs＜3.10%）. Cluster analysis， PCA， and OPLS-DA revealed that C. sinensis Osb. and C. sinensis were clustered into one category， while C. aurantium，C. junos and C. aurantium Linn. were clustered into another category. The variable importance projection values for neohesperidin， auraptene， naringin， neoponcirin， tangeretin， hesperidin， sinensetin， and 5，7-dihydroxychromone-7-neohesperidoside were all greater than 1. CONCLUSIONS In this study， the contents of 15 components in Aurantii Fructus Immaturus from different origins are determined， and 8 differential components， including neohesperidin， auraptene， naringin， and neoponcirin， are screened out.

Establishment of rat models of liver transplantation provides an ideal animal model for resolving the problems of postoperative complications and perioperative treatment of liver transplantation. With in-depth study of the establishment of rat models of liver transplantation, classic "two-cuff" technique has been gradually employed. However, poor surgical field, vascular torsion, biliary tract injury and long anhepatic phase remain unresolved in the process of liver transplantation using traditional techniques. At present, the rat models of liver transplantation at home and abroad are modified mainly from the reconstruction of four vital anatomic structures including the suprahepatic inferior vena cava, portal vein, infrahepatic inferior vena cava and bile duct. Therefore, the latest progress in the reconstruction of the suprahepatic inferior vena cava, portal vein, infrahepatic inferior vena cava and bile duct was reviewed, aiming to provide reference for the establishment of rat models of liver transplantation and promote further development of liver transplantation techniques.

Objective To evaluate the role and potential mechanism of interleukin (IL)-18/IL-18 binding protein (BP) in mediating the killing effect of natural killer (NK)-92MI cells upon endothelial cells from α-1, 3- galactosyltransferase gene-knockout (GTKO) porcine models. Methods NK-92MI cells were divided into the NK, NK+IL-18, NK+GTKO, IL-18+NK+GTKO and IL-18+IL-18BP+NK+GTKO groups. The messenger ribonucleic acid (mRNA) levels of inflammation-related genes in NK-92MI cells were detected by quantitative real-time reverse transcription polymerase chain reaction (qRT-PCR). The killing effect of NK-92MI cells on endothelial cells from GTKO porcine models was evaluated by lactate dehydrogenase (LDH) assay. The apoptosis of endothelial cells from GTKO porcine models was assessed by terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) assay. The expression levels of proteins with killing effect and apoptosis-related proteins were determined by Western blot. Results Compared with the NK, NK+IL-18 and NK+GTKO groups, the expression levels of interferon (IFN)-γ, tumor necrosis factor (TNF)-α, IL-8, IL-3, IL-6 and granulocyte-macrophage colony stimulating factor (GM-CSF) mRNA were up-regulated in NK-92MI cells in the IL-18+NK+GTKO group, and the differences were statistically significant (all P < 0.05). Compared with the IL-18+NK+GTKO group, the expression levels of IFN-γ, TNF-α, IL-8, IL-3, IL-6 and GM-CSF mRNA were down-regulated in NK-92MI cells in the IL-18+IL-18BP+NK+GTKO group, and the differences were statistically significant (all P < 0.05). Compared with the NK+GTKO group, the expression levels of perforin, granzyme B and IFN-γ proteins in NK-92MI cells were up-regulated, the killing rate of NK-92MI cells against endothelial cells from GTKO porcine models was enhanced, the apoptosis rate of endothelial cells from GTKO porcine models was increased, and the ratios of B cell lymphoma-2 (Bcl-2)-associated X protein (Bax)/Bcl-2 and cleaved Caspase-3/Caspase-3 in endothelial cells from GTKO porcine models were elevated in the IL-18+NK+GTKO group, and the differences were statistically significant (all P < 0.05). Compared with the IL-18+NK+GTKO group, the expression levels of perforin, granzyme B and IFN-γ proteins were down-regulated, the killing rate of NK-92MI cells against endothelial cells from GTKO porcine models was decreased, the apoptosis rate of endothelial cells from GTKO porcine models was decreased, and the ratios of Bax/Bcl-2 and cleaved Caspase-3/Caspase-3 in endothelial cells from GTKO porcine models were declined in the IL-18+IL-18BP+NK+GTKO group, and the differences were statistically significant (all P < 0.05). Conclusions IL-18BP may block the expression of inflammation-related genes in NK-92MI cells induced by IL-18 and the killing effect of NK-92MI cells on endothelial cells from GTKO porcine models.

Purpose: This study attempted to detect the changes of cervical cancer screening rate and willingness among female migrants, and the associated socio-demographic factors in Shenzhen city. Materials and Methods: Two citywide surveys were conducted using a multistage random cluster sampling method in 2011 and 2014, respectively. Data on demographic characteristics, screening participation, and willingness to screen were collected. Logistic regression models were applied to detect possible associated socio-demographic characteristics, and their variations with survey years. Results: In total, 12,017 female migrants were enrolled, with a mean age (standard deviation) of 36.73 (6.55) years. From 2011 to 2014, the screening rate increased (25.8% vs. 35.1%, p < 0.001), while the willingness to screen remained stable (82.2% vs. 82.8%, p=0.46). Overall, socio-demographic characteristics of female migrants, including age, marital status, education, monthly income, employment, and medical insurance, were found to be positively associated with screening participation. Similar impacts in relation to willingness were observed except for age. However, these associations varied with survey years, mainly in the contributions of education and monthly income to screening participation, as well as age, monthly income, and medical insurance to willingness of being screened. Conclusion Identifying changes of associated socio-demographic factors precisely is warranted of necessity, which provides novel clues to adjust targeted actions regularly in promoting cervical cancer screening participation among female migrants in Shenzhen.

DNA methylation as an important aspect of epigenetics plays an important role in spermatogenesis and embryonic development. In recent years, researchers have found that male infertility, in particular abnormal semen quality, is related to abnormal DNA methylation. To further delineate the pathogenesis of male infertility and inspire new ideas for the treatment of male infertility, a comprehensive review over the correlation between abnormal methylation of imprinted genes, repetitive DNA elements and non-imprinted genes, semen quality (including sperm count, morphology, and vitality) and male infertility is provided.
DNA Methylation ; Humans ; Infertility, Male/genetics* ; Male ; Semen Analysis ; Sperm Count ; Spermatogenesis ; Spermatozoa/pathology*

4,6-α-glucosyltransferases (4,6-α-GTs), which converts amylose into α(1-6) bonds-containing α-glucan, possesses great application potential in enzymatic synthesis of dietary fiber. Primers were designed according to the conserved motifs existing in the amino acid sequence of 4,6-α-GTs, and used to amplify a putative GTFB-Like 4,6-α-GTs gene (named as gtf16) from the genomic DNA of Lactobacillus. The gtf16 gene was cloned into the plasmid pET15b, expressed in Escherichia coli BL21(DE3), followed by purification and characterization. The optimum pH and the optimum temperature of the purified enzyme were 5.0 and 40 °C, respectively. The biotransformation product of this enzyme was systematically characterized by thin-layer chromatography, NMR spectroscopy, and hydrolysis reaction. The Gtf16-catalyzed product shows a similar structure to that of the isomalto/malto-polysaccharide (IMMP), which is the amylose-derived product catalyzed by GtfB from Lactobacillus reuteri 121. Moreover, The Gtf16-catalyzed product contains up to 75% of α(1-6) bonds and has an average molecular weight of 23 793 Da. Furthermore, the content of the anti-digestive components was 88.22% upon hydrolysis with digestive enzymes.
Bacterial Proteins/genetics* ; Glucans ; Glucosyltransferases/genetics* ; Lactobacillus fermentum/enzymology*

Objective:To investigate whether adjuvant skin-marker positioning can decrease the set-up errors in overweight patients with thoracic and abdominal tumors.Methods:A total of 60 overweight patients with thoracic and abdominal tumors treated with radiotherapy in the First Affiliated of Fujian Medical University between January 2018 and December 2018 were randomly divided into two groups. In group A, conventional skin-marker positioning was adopted. In group B, conventional skin-marker positioning combined with adjuvant skin-marker position was employed. All patients were immobilized with thermoplastic positioning body membrane with head-body plate fixation. The set-up errors in the right-left, head-foot and dorsoventral directions were obtained from cone-beam CT (CBCT) scan system before radiation delivery. The set-up errors were statistically compared between two groups by using t-test. Results:In group A, the set-up errors in the right-left, head-foot and dorsoventral directions were (4.47±2.91) mm, (5.43±2.61) mm and (3.87±2.40) mm, significantly higher compared with (2.97±1.68) mm, (3.21±1.62) mm and (2.59±1.57) mm, respectively (all P<0.001). Conclusion:Adjuvant skin-marker positioning method can reduce the set-up errors and enhance the positioning repeatability in overweight patients with thoracic and abdominal tumors receiving radiotherapy.

Objective: To explore the association between screen time and autistic behavior in infants and young children, and to provide clues to the mechanism for further research. Methods: The primary caregivers of 22 586 children in the district of Longhua in Shenzhen were surveyed. Demographic data and screen time were collected using a selfdisigned questionnaire, and children’s autistic behavior was assessed using the child Autism Behavior Checklist(ABC).The chisquare test was used to analyze the correlation between demographic data and screen time, demographic data and autistic behavior, and screen time and autistic behavior. Unconditioned Logistic regression model was used to study the effect of electronic screen exposure on autistic behavior. Results: The amount of screen time spent in infancy increased with age. For children ages 0-,1- and 2-3 years, 60.1%, 35.0% and 20.2% respectively did not watch TV, and 74.0%, 52.6% and 26.8% respectively did not watch the new generation of electronic products. The positive rate of ABC scale screening was 5.3%, including 6.0% male and 4.4% female, OR(95%CI)=1.37 (1.23-1.54).There was correlations between screen time and autistic behavior in infants at all ages (P<0.05).Screen time increased the risk of autistic behavior in younger age groups than in older age groups. For 1 year olds with moderate screen exposure, increased screen exposure at 2 to 3 years of age was associated with an increased risk of autistic behaviors, while reduced screen exposure at 2 to 3 years of age was associated with a lower risk of autistic behaviors(OR=2.14, 2.77, P<0.05). The higher daily screen time at 0-3 years old was, the greater risk of autistic behaviors. Compared with the noncontact electronic screen group, the OR values of the daily TV screens in the ≥1 h/d group and the ＜1 h/d group were 2.01 and 2.45, respectively (P＜0.05).Compared with the non-contact electronic screen group, the OR values of the screens exposed to the new generation of electronic products in the ≥1 h/d group and the ＜1 h/d group were 2.01 and 2.33, respectively(P<0.05).The higher the time of single exposure to electronic screen between 0 and 3 years old, the greater the risk of autistic behaviors. The OR values were 2.50, 1.79 and 1.47 when ≥1 h/time, 0.5-1 h/time and 15-30 min/time compared with <15 min/time（P<0.05）. Conclusion Early exposure to electronic screens in infants, excessive total daily exposure to electronic screens, and excessive screen time each time are all likely to increase autistic behavior. Therefore, it is suggested that children under 2 years old should not be exposed to electronic screens every day. Children aged 2-3 years old who are exposed to electronic screens＜0.5 h/d and whose screen time <15 min might not significantly increase autistic behaviors.

OBJECTIVE： To provide scientific basis for the utilization and development of Miao medicine Oxalis corniculata by promoting the quality standard of it. METHODS： Total of 12 batches of O. corniculata were collected from Guizhou， Anhui and Henan， etc. Microscopic characteristics of 12 batches of O. corniculata powder were observed. According to the corresponding methods in 2015 edition of Chinese Pharmacopoeia （part Ⅳ）， TLC was used for qualitative identification [developing solvent was trichloromethane-methanol-formic acid （8 ∶ 1 ∶ 0.1， V/V/V）]， and the contents of moisture， total ash， acid insoluble ash and alcohol soluble extractive from 12 batches of O. corniculata were determined. The content of isovitexin was determined by HPLC. The determination was performed on Venusil XBP C18 （L） with mobile phase consisted of acetonitrile-0.1％ phosphoric acid solution  （15 ∶ 85， V/V） at the flow rate of 1 mL/min. The column temperature was 35 ℃， and the detection wavelength was set at 338 nm. The sample size was 10 μL. RESULTS： Microscopic observation showed that the powder was grayish brown to yellowish brown， with many non-glandular hairs and obvious fibrous pore. Results of TLC identification showed that the spots of the same color appeared in the corresponding positions of the test and the control chromatogram. The contents of moisture， total ash， acid insoluble ash and alcohol soluble extract from samples were 6.66％-12.13％， 9.16％-13.79％， 1.58％-4.63％ and 5.22％-15.79％， respectively. Results of HPLC method showed that the concentration of isovitexin showed a good linear relationship in the range of 5.20-78.3 μg/mL （r＝0.999 0）； RSDs of reproducibility （n＝9）， intermediate precision （n＝6） and stability （24 h， n＝6） tests were all lower than 2.0％； and the recovery rates were 97.54％-99.52％ （RSD＝0.74％， n＝6）； the contents of isovitexin in 12 batches of O. corniculata were 0.036％-0.144％ （n＝3）. CONCLUSIONS： Qualitative and quantitative identification methods of O. corniculate were established， which can be used as a reference for improving the quality standard of O. corniculata.

Objective To evaluate the diagnostic value of the early rheumatoid arthritis (RA) classification criteria for early RA and to assess the clinical efficacy of knee arthroscopic synovectomy for RA.Methods A total of 62 patients who had arthritic complaints with disease duration less than one year were treated with knee arthroscopic synovectomy.Patients were diagnosed as RA confirmed by changes of synoviomorphous under arthroscopy,synovial biopsy,immunological biochemical laboratory and MRI,and compared with the preoperative ERA diagnosis.The efficacy was assessed at the third months and more than 12 months after operation,including Health Assessment Questionnaire (HAQ),Lysholm score,laboratory parameters of erythrocyte sedi-mentation rate (ESR) and C-reactive protein (CRP).Results The sensitivity and specificity in the early RA classification criteria for RA diagnosis were 81.58％ and 91.67％.38 patients diagnosed as RA and the remaining 24 patients were significantly improved of HAQ,Lysholm score,ESR,CRP compared with preoperation,showed statistical difference (P ＜ 0.01).There were still statistical differences between the follow-up results of the group of patients with RA in different period after operation (P ＜ 0.01).Conclusion The early RA classification criteria is worthy of clinical application,because it has the advantages of simple,practical,and is beneficial to early diagnosis of RA.The early RA patients should be treated with knee arthroscopic synovectomy as far as possible.It can improve not only the knee function,but also the general symptoms,and it can interrupt the RA pathological process,effective maintain curative effect after operation.