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Continuous exposure of oocytes to elevated concentrations of insulin compromises embryonic developmental competence. However, the effects of insulin on oogenesis from fetal germ cells are unknown. The objective of this study was to assess the effect of continuous insulin exposure, with or without FSH, on oogenesis and follicular development. A simple and efficient method was established that could be used to obtain oocytes from pre-meiotic germ cells in 12.5 days post-coitum (dpc) fetal mouse ovaries using a three-dimensional culture system with serum-free medium. Mouse 12.5 dpc fetal ovaries were cultured for 14 days with or without insulin/FSH. Low (0.2–1 μg/ml) or high (5–20 μg/ml) doses of insulin retarded oocyte growth in vitro. Insulin at 5 lg/ml led to significant oocyte growth retardation (P < 0.05), while FSH alleviated the deleterious effect of insulin. Most importantly, the proportion of secondary follicles at 12 days post-culture in the presence of insulin was reduced significantly compared with controls (P < 0.05). Expression levels of genes specific for ovarian cells, e.g. Cx37, Cx43, Scp3, Bax and FSHR, were significantly reduced when exposed to insulin during oogenesis (P < 0.05). The data suggest that insulin has a profound detrimental effect on oogenesis and folliculogenesis in vitro. [ABSTRACT FROM AUTHOR]

A systematic investigation of D3 C32 fullerene and its derivatives C32Xn (X = H and Cl) has been performed using B3LYP/6-31G(d) method based on the density functional theory. The geometry structures, reaction energies, relative stabilities, and electronic properties have been studied. By investigating the possible C32Xn (X = H and Cl) molecules, C32H2 and C32Cl2 behave more thermodynamically accessible with respect to other derivatives. The frontier molecular orbitals and electronic density of states calculations of C32X2 system indicate that H and Cl passivation have less contribution to the electronic structures, but significantly improve the stability of D3 C32 fullerene. Finally, the 13C NMR chemical shifts of C32H2 and C32Cl2 have been simulated to provide helpful information for further experiment identification. [ABSTRACT FROM AUTHOR]

Frequency-selective surfaces (FSSs) with two sharp sidebands have been investigated, which are constructed by cascading and shunting two substrate integrated waveguide cavities (SIWCs) of different sizes with periodic slots at top and bottom metal surfaces. The lower and upper sharp sidebands are induced by two transmission nulls that are generated by coupling between the slot aperture resonance and the resonances in the two cavities. Samples of the shunted SIWC-FSS with different size cavities were fabricated. Measured results are in good agreement with the predicted ones and its performance, such as high selectivity and passband bandwidth, is stable at oblique incidence. [ABSTRACT FROM AUTHOR]

Planar shock compression effects on void formation and cracking in Zr41Ti14Cu12.5Ni10Be22.5 bulk metallic glass (BMG) are studied in this paper. Cracking was found to be a result of void linkage in some direction deviation from the maximum shear stress plane. Changing the state of the stress inside the BMG sample led to formation of different void distribution. Nucleation of the microvoids was possibly initiated by release of excess free volume under shock wave compression. [ABSTRACT FROM AUTHOR]

Structural relaxations of a Pd[sub 39]Ni[sub 10]Cu[sub 30]P[sub 21] bulk metallic glass (BMG) under ambient pressure and at a high pressure are compared by means of temperature-modulated differential scanning calorimetry (TMDSC) measurements. The structural relaxation is evaluated by the enthalpy recovery involved upon passing through the glass transition in the irreversible TMDSC curves. It is found that the isothermal relaxation of the BMG under ambient pressure leads to a lower-enthalpy state than that under a high pressure. The onset temperature for the enthalpy recovery process is found to exhibit a linear dependence on the recovery enthalpy. [ABSTRACT FROM AUTHOR]

Short-range structure of a Zr41Ti14Cu12.5Ni10Be22.5 bulk metallic glass prepared by shock-wave treatment was investigated by x-ray diffraction using synchrotron radiation with a wavelength of 0.112 71 Å. The radial distribution function was obtained from S(Q) with a large Q value up to 20 Å-1. The oscillation in S(Q) of the glass definitely persists up to Q∼14 Å-1. The shoulder on the high Q side of the second peak is observed in the bulk glass. It is found that the glass has higher coordination numbers in the range of r∼2.4–5.6 Å and lower numbers in the range of r∼5.6–9.5 Å than those for a water-quenched glass while the shell distances are similar in both glasses prepared by shock-wave and water-quenching methods. In the shock-waved glass, atomic configurations in the first, fourth, or fifth coordination shells are modified, i.e., atoms are packed denser in the first two coordination shells and less (or more free volume) in the third and fourth coordination shells as compared to those for the water-quenched glass. This atomic configuration for shock-waved glass might have different physical properties. © 2004 American Institute of Physics. [ABSTRACT FROM AUTHOR]

Summary: The paper reports the length–weight relationships of 3 species from the Poyang Lake, the largest freshwater lake freely connect to the Yangtze River, China. A total of 1,207 specimens were collected seasonally (January, April, September and November, 2010) using various types of fishing gear nets (mesh size: 1–12 cm) and stationary lift nets (mesh size: 0.7–4.4 cm). For each species, the sample size, length range, weight range, LWR, 95% confidence intervals of a and b, and coefficient of correlation were determined. Length‐weight relationships for 1 species were unknown to FishBase, and new maximum lengths are recorded for 2 species. [ABSTRACT FROM AUTHOR]

Here the authors report that by a high pressure synthesis method, superconductivity with an onset transition temperature at 52 K was discovered in a fluorine-doped quaternary iron-arsenide compound Pr[O1-xFx]FeAs, with the zero resistivity and Meissner transition appeared at 44 K and 50 K respectively. As the first non-cuprate compound that superconducts above 50 K, this discovery places these iron arsenide compounds to the second high temperature superconducting family explicitly. [ABSTRACT FROM AUTHOR]

Without any perturbations on the surface of the patch resonator or orthogonal feed lines, a new microstrip dual-mode triangular-patch bandpass filter with low insertion loss and one transmission zero is presented. The degenerate modes are realised by use of the input/output spur-lines. It has been verified by simulation and measurement. [ABSTRACT FROM AUTHOR]

A new dual-spiral-shaped slot resonator in a coplanar waveguide (CPW-DSR) is studied. Also, its equivalent circuit model is introduced and used for multi-resonant performance analysis. Finally, a bandstop filter using periodically loaded CPW-DSRs is presented and verified by simulations and measurement. [ABSTRACT FROM AUTHOR]

Replies to the criticism on reversible phase transition between amorphous and crystalline in Zr[sub 41.2]Ti[sub 13.8]Cu[sub 12.5]Ni[sub 10]Be[sub 22.5] under high pressure at room temperature. Adoption of large scattering angle in the study; Characteristics of the transition behavior of the bulk metallic glass; Influence of photon energy on absorption.

The pressure-dependent acoustic velocities of a Pd[sub 39]Ni[sub 10]Cu[sub 30]P[sub 21] bulk metallic glass (BMG) have been measured up to 0.5 GPa by using an ultrasonic technique with the pulse echo overlap method. The elastic constants, the Debye temperature, and their pressure dependence are obtained. The isothermal equation of state (EOS) of the BMG is established in terms of the Murnaghan form. The atomic configurations of the BMG are discussed by comparing the elastic constants and the EOS with those of its metallic component and of other amorphous materials. © 2000 American Institute of Physics. [ABSTRACT FROM AUTHOR]

Acoustic velocities of Pd[sub 39]Ni[sub 10]Cu[sub 30]P[sub 21] bulk metallic glass (BMG) are measured by an ultrasonic technique upon annealing. The elastic constants and the Debye temperature are obtained. A large softening of the transverse phonon is exhibited in the as-quenched BMG relative to its crystallized state. Upon crystallization, the shear modulus and the Debye temperature increase by ∼30% and ∼12%, respectively; however, the density increases by only ∼0.6%. Some anomalous acoustic and elastic behaviors are observed near the glass transition temperature and in the supercooled liquid region of the BMG. The anomalies are explained with regard to the structural changes. © 2000 American Institute of Physics. [ABSTRACT FROM AUTHOR]

The potential benefits of supplementing pyrroloquinoline quinone disodium (PQQ·Na2) in the diet of broiler chicks were explored. We first examined the effect of different levels of dietary PQQ·Na2 on growth performance, carcass characteristics, and plasma biochemical parameters (trial 1). A total of 490 1-day-old male Arbor Acres (AA) broiler chicks were randomly divided into 5 dietary groups supplemented with 0, 0.05, 0.1, 0.2, or 0.4 mg PQQ·Na2/kg feed. As the 0.2 mg/kg PQQ·Na2 supplement gave the best performance, we then investigated whether this level of PQQ·Na2 influenced the redox status of plasma samples and mitochondrial-related metabolism (trial 2). A total of 120 1-day-old male AA chicks were randomly divided into 2 groups supplemented with 0 or 0.2 mg PQQ·Na2/kg diet. In trial 1, birds fed a diet containing 0.2 mg PQQ·Na2/kg showed lower feed conversion ratio compared with those fed the control diet in the overall study (d 1 to 42, P = 0.039). Breast muscle yield (d 42) increased quadratically in response to dietary PQQ·Na2 supplementation (P = 0.021). Analysis of plasma biochemical parameters revealed that feeding broiler chicks with ≤ 0.4 mg/kg PQQ·Na2 did not cause adverse health effects. In trial 2, birds fed 0.2 mg/kg PQQ·Na2 again showed improved feed efficiency than the control birds in the grower and overall phases (P = 0.038 and 0.016, respectively). In addition, dietary PQQ·Na2 supplementation resulted in a higher anti-oxidative capacity (P = 0.001), lower redox potential (P = 0.008), and higher hepatic citrate synthase activity (P = 0.002). In contrast, no difference in hepatic mitochondrial DNA copy number was observed between the 2 experimental groups (P > 0.1). These results indicate that PQQ·Na2 is a potentially effective feed additive for improving feed efficiency, stimulating breast muscle development, and maintaining redox status in broiler chicks. Enhancement of mitochondria efficiency, rather than modulating mitochondria numbers, may underlie the growth-promoting effect of PQQ·Na2. [ABSTRACT FROM AUTHOR]

Ameloblastoma (AM) is a benign odontogenic tumor with unknown etiology. It is prone to recurrence and has a potential for malignant transformation. Patients often show high rates of relapse after curettage, or suffer from structural and functional damage of jaw after partial resection. Whole-genome sequencing data revealed that BRAF mutations and SMO mutations were common and likely to be mutually exclusive in AM. It was also reported that BRAF inhibitors were effective in several patients carrying BRAF V600E mutation. However, reliable preclinical models are urgently needed for exploring targeted therapy as it's so difficult to conduct large clinical trials in this tumor. Patient-derived cell models in vitro and xenograft models in vivo are frequently used preclinical models. In fact, benign tumor cells generally showed a finite proliferative capacity in two-dimensional culture, and most likely, they could exhibit altered cellular phenotype after immortalization. Moreover, this benign tumor presented low chances of subcutaneous engraftment in nude mice. Accordingly, humanized mouse xenograft model needs more exploration. Yet, it is worth mentioning that a three-dimensional organoid model presents a high potential in culturing stem-cell-like epithelial cells in AM, and it would further be used in recapitulating corresponding tumors and developing targeted medicines. In this paper, we review research progress in preclinical models and the genetic variations of AM, and raise drug screening prospect of the current organoid models, which may pave the way for the possible personalized medicine in AM.

Objective: To explore the characteristics of viral infections in children with diarrhea in Beijing from 2018 to 2022. Methods: Real-time PCR and enzyme-linked immunosorbent assay were used to detect viral nucleic acid of Norovirus (NoV), Sappovirus (SaV), Astrovirus (AstV), Enteric Adenovirus (AdV) or antigen of Rotavirus (RV) in 748 stool samples collected from Beijing Capital Institute of Pediatrics from January 2018 to December 2021. Subsequently, the reverse transcription PCR or PCR method was used to amplify the target gene of the positive samples after the initial screening, followed by sequencing, genotyping and evolution analysis, so as to obtain the characteristics of these viruses. Phylogenetic analysis was performed using Mega 6.0. Results: From 2018 to 2021, the overall detection rate of the above five common viruses was 37.6%(281/748)in children under 5 years old in Beijing. NoV, Enteric AdV and RV were still the top three diarrhea-related viruses, followed by AstV and SaV, accounting for 41.6%, 29.2%, 27.8%, 8.9% and 7.5%, respectively. The detection rate of co-infections with two or three diarrhea-related viruses was 4.7% (35/748). From the perspective of annual distribution, the detection rate of Enteric AdV was the highest in 2021, while NoV was predominant in the other 4 years. From the perspective of genetic characteristics, NoV was predominant by GⅡ.4, and after the first detection of GⅡ.4[P16] in 2020, it occupied the first two gene groups together with GⅡ.4[P31]. Although the predominant RV was G9P[8], the rare epidemic strain G8P[8] was first detected in 2021. The predominant genotypes of Enteric AdV and AstV were Ad41 and HAstV-1. SaV was sporadic spread with a low detection rate. Conclusion: Among the diarrhea-related viruses infected children under 5 years of age in Beijing, the predominant strains of NoV and RV have changed and new sub-genotypes have been detected for the first time, while the predominant strains of AstV and Enteric AdV are relatively stable.

Dairy products are the major source of odd- and branched-chain fatty acids (OBCFAs), a group of nutrients with emerging health benefits. The animal diet is known to influence milk fat OBCFAs of dairy cows; however, little is known about the effects of physiological factors. The objective of this study was to investigate the effects of parity and lactation stage on OBCFAs in milk fat of dairy cows. Holstein dairy cows (n = 157) were selected according to parity (first, second, third, or greater) and days in milk (DIM) (≤21 DIM, 21 < DIM ≤ 100, 100 < DIM ≤ 200, >200 DIM). All cows were fed the same total mixed ration for three weeks. Milk samples were collected during the last three days of each lactation stage for fatty acid (FA) analyses via gas chromatography. Results showed that first- and second-parity cows displayed significantly higher proportions and yields of iso-14:0, iso-15:0, iso-16:0, total iso-FA, and total branched-chain FA (P < 0.05) compared with other parities. The proportions of C17:0 and C17:1 cis-9 were also greater in first-parity cows (P < 0.05), while the yields of C17:0 and C17:1 cis-9 were similar among different parities (P > 0.05). The proportions of total OBCFAs were greater in first- and second-parity cows (P < 0.05), whereas the highest yield was observed in second-parity cows. Lactation dairy cows in ≤ 21 DIM group displayed lower proportions of iso-13:0, anteiso-13:0, C13:0, iso-14:0, C15:0, iso-16:0, total iso-FA, and total OBCFAs compared with that of the other groups (P < 0.05), and also lower yields of iso-14:0 and iso-16:0 (P < 0.05). In contrast, C17:0 and C17:1 cis-9 proportions and yields were higher in dairy cows with ≤ 21 DIM (P < 0.05). Iso-17:0 and anteiso-17:0 were not affected by lactation stage (P > 0.05). Taken together, our data showed that both parity and lactation stage have considerable effects on milk fat OBCFAs of dairy cows. In summary, first- and second-parity cows had higher milk OBCFAs compared with later parity cows, and OBCFAs with medium chain lengths were lower in dairy cows with ≤ 21 DIM, while C17:0 and C17:1 cis-9 were higher. These findings show that milk OBCFA contents are differentially modulated by physiological state. They will be useful in future studies that seek to alter OBCFA composition of Holstein dairy cow milk fats., (Copyright © 2022 The Authors. Published by Elsevier B.V. All rights reserved.)

Objective: To investigate the effect of berberine on programmed necrosis of hepatocytes induced by metabolic-associated fatty liver disease (MAFLD) in mice and its related molecular mechanism. Methods: Twenty male C57BL/6N mice were randomly divided into four groups (n=5 in each group): control group (S), fatty liver group (H), berberine group(B), nuclear factor erythroid 2-related factor 2 inhibitor group (Nrf2), and all-trans-retinoic acid (ATRA) group (A). Serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), lactate dehydrogenase (LDH), triglycerides (TG), total cholesterol (TC), tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β) concentrations were detected at the end of week 12 to calculate fatty liver index (liver mass/body mass ratio). Liver tissue was stained with HE, Masson and Oil Red O, and SAF score was used to evaluate the degree of liver injury. The expression levels of hepatic programmed necrosis-related proteins, namely receptor-interacting protein kinase 3 (RIPK3), phosphorylated mixed series protease-like domain (p-MLKL) and Nrf2 were detected by Western blot method. One-way ANOVA was used for intragroup comparisons and LSD-t tests were used for intergroup comparisons. Results: Compared with S group, H group serum ALT, AST, LDH, TG, TC, TNF-α, IL-1β levels and fatty liver index were significantly increased. The liver tissue was filled with vacuolar-like changes and inflammatory cell infiltration. Numerous red lipid droplets were observed with oil red O staining. Collagen fiber hyperplasia was evident with Masson staining. SAF scores (6.60 ± 0.55 and 0.80 ± 0.45) were significantly increased. The expressions of RIPK3 and p-MLKL were up-regulated. Nrf2 level was relatively increased, and the differences were statistically significant ( P < 0.05). Compared with H group, berberine intervention group liver biochemical indexes, lipid levels, pro-inflammatory mediator expression, fatty liver index, and SAF score were significantly reduced, and the expression of RIPK3 and p-MLKL were down-regulated, while Nrf2 levels were further increased, and the differences were statistically significant (P<0.05). Compared with B group, treatment with Nrf2 inhibitor had antagonized the protective effect of berberine on fatty liver. Serum ALT, AST, LDH, TG, TC and TNF-α, IL-1β levels, fatty liver index, and SAF scores were significantly increased and the expressions of RIPK3 and p-MLKL were relatively increased, and the differences were statistically significant ( P < 0.05). Conclusion: Berberine can significantly improve the metabolic-associated fatty liver disease injury in mice, and its mechanism is related to activation of Nrf2 and inhibition of programmed necrosis of hepatocytes.

Objective: To analyze the distribution and genetic characteristics of sporadic adult diarrhea virus in Chaoyang District, Beijing. Methods: Fecal samples from 177 adult patients with sporadic diarrhea were collected from 4 enteric outpatient clinics in Chaoyang District, Beijing from May to December 2019. Nucleic acid detection of Norovirus, Sappovirus, Rotavirus, Enteric Adenovirus and Astrovirus in the samples was performed by real-time quantitative PCR. The positive samples were amplified by RT-PCR/PCR and sequenced. The phylogenetic analysis was performed by neighbor-Joining (NJ) methods of Mega 6.0 software. Results: There were 60 of 177 (33.90%) adult sporadic diarrhea samples positive for enteric viral pathogens. Among them, 47 cases were infected with single virus, including 29 cases of Norovirus, 9 cases of Sappovirus, 8 cases of Astrovirus and 1 case of Enteric Adenovirus, in addition with 13 cases of multiple infections. None of rotavirus was detected. Partial sequences were successfully obtained for analysis, including 16 cases of GI Norovirus (7 subtypes and GI.3[P13] predominant), 10 cases of GII Norovirus (5 subtypes and GII.6[P7] predominant), 12 cases of Sappovirus (4 subtypes and GI.2 predominant), and 7 cases of Astrovirus (2 subtypes and AST-1 predominant). Conclusion: Norovirus, Astrovirus and Sappovirus are main pathogens among sporadic adult diarrhea in Beijing in 2019, and and different pathogenic gene subtypes show diverse characteristics.

Objective: To investigate the research trend and scope of prevention of central venous catheter-related bloodstream infection (CRBSI) in burn patients. Methods: The scoping review method was adopted. Pre-retrieval was carried out with search terms of ", , " and "central venous catheter, infection, catheter-related bloodstream infection, burn". On the basis of pre-retrieval, different retrieval formulas were formulated to retrieve researches related to central venous CRBSI in burn patients in China National Knowledge Internet , Wanfang Database , VIP Database , PubMed , Embase , CINAHL , and Cochrane Library from the establishment of each database to August 2020. Data were extracted from the included literature, including the first author, research publication time, research country, research type, diagnosis basis and intervention measures of central venous CRBSI, research sample selection, incidence related to infection, and research conclusion. Results: A total of 20 randomized controlled trials, quasi-experimental studies, case-control studies, cohort studies, and implementation researches published in 1990-2020 were included in this study with the first authors from China, the United States of America, or Argentina. The diagnostic bases for central venous CRBSI in burn patients were not uniform in the included literature, including adopting the Guidelines of American Centers for Disease Control and Prevention, Diagnostic Criteria for Nosocomial Infection , and other diagnostic criteria without specifying the source. The intervention measures included the use of new materials such as antibiotics coated catheter and ethanol impregnated port protectors, multidisciplinary cooperation, and comprehensive preventive measures. The sample size in the included literature was small, and the sample selection was different, including the number of patients and the the number of placement of central venous catheter. The outcome indicators for infection in the included literature were diversified. The incidence per 1 000 days of central venous CRBSI was 20.41‰-29.1‰ of patients in control group in China, the incidence per 1 000 days of central venous CRBSI was mostly <16.6‰ in control group in foreign countries, and the incidence of central venous CRBSI was decreased to varying degrees after implementing the corresponding intervention measures. Related research conclusions showed that new materials, multidisciplinary cooperation, and comprehensive preventive measures had good effects on prevention of central venous CRBSI in burn patients. Conclusions: The researches on prevention of central venous CRBSI in burn patients in China start early and the research types are diversified. The diagnostic criteria of central venous CRBSI in burn patients are not uniform, intervention measures have shifted from standardizing relevant operational measures to exploring the prevention effects of new materials, multidisciplinary cooperation, and multiple measures, and the latter has good effects on preventing central venous CRBSI in burn patients.

Objective: To explore the influencing factors on perinatal mortality of pregnant women with HIV infection to reduce the mother-to-child transmission in Sichuan province. Methods: In this study, 4 786 perinatal infants of the HIV-infected pregnant women were included. Related data on perinatal epidemiology was reported by all the 183 medical and health care institutions where the HIV prevention of mother-to-child transmission program was initiated in 2005-2016. Univariate χ (2) test and multivariate logistic regression methods were used to analyze the perinatal mortality outcomes and influencing factors. Results: The overall perinatal mortality rate was 25.7‰ (123/4 786) among HIV-infected pregnant women, with annual downwarding trend (trend χ (2)=32.220, P =0.000). Perinatal mortality rate appeared the highest ( χ (2)=4.130, P =0.042), with more fetal deaths and stillbirths and less early neonatal death within 7 days in Liangshan county ( χ (2)=29.626, P =0.000). Results from the multivariate logistic regression analysis showed that fewer pregnant numbers would contribute to the, lower perinatal mortality rate (1-2 pregnancies OR =0.417, 95 %CI : 0.184-0.943; 3-4 pregnancies OR =0.447, 95 %CI : 0.223-0.895). Perinatal deaths were more likely to be prevented if LPV/r protease inhibitor-based triple antiviral therapy was provided ( OR =0.530, 95 %CI : 0.285- 0.986) or delivery was taken place in the hospital (hospital of municipal-level and above OR =0.222, 95 %CI :0.098-0.499; county-level hospital OR =0.282, 95 %CI : 0.166-0.480; township-level hospital OR =0.134, 95 %CI : 0.031-0.586) among HIV-infected pregnant women. However, premature delivery or neonatal asphyxia would increase the risk of perinatal mortality (premature delivery OR =8.285, 95 %CI : 5.073-13.533; neonatal asphyxia OR =9.624, 95 %CI : 4.625-20.028). Conclusions: The perinatal mortality rate of HIV-infected pregnant women appeared significantly higher than that in the province or the whole country. Strategies involving LPV/r-based triple antiviral therapy, promotion of hospital delivery, reducing the incidence rates of premature deliveries and neonatal asphyxia, should be strengthened.

The Wilderness Medical Society updated and published the Wilderness Medical Society practice guidelines for the prevention and treatment of frostbite in July 2019. The guidelines provide evidence-based recommendations for the prevention and treatment of frostbite, mainly including pathophysiology, classification, prevention, and treatment of frostbite. This paper focuses on the interpretation of the guidelines, aiming at making clinical medical staff understand the new progress of frostbite treatment and providing reference for clinical practice.

Objective: To quantitate the association between birth weight and phenotypes of physical indicators in adulthood, i.e. BMI and waist circumference (WC) and to what degree genetic or environmental factors affect birth weight-obesity association. Methods: A total of 6 623 gender matched twin pairs aged 25 to 79 years were recruited through the Chinese National Twin Registry. The twins reported their own birth weight, current height and weight, and WC using a self-administered questionnaire. BMI was calculated according to the self-reports of body height and weight. Within twin-pair design was used to quantitate the association between birth weight and phenotypes related to obesity while bivariate structural equation models were used to decompose the phenotype correlation. Results: After adjusted for multiple factors, twin-pair analyses within monozygotic (MZ) showed that, on average, a 1.0 kg increase in birth weight corresponded to an increase of 0.33 kg/m(2) in BMI and 0.95 cm in WC in adulthood ( P <0.001). Bivariate structural equation models showed significant positive unique environmental correlation between birth weight and the two obesity-related phenotypes. Conclusion: The study supported the role of twin-specific supply line factors on relationship between birth weight and physical indicators in adulthood.

Objective: To explore chromobox protein homolog 2 (CBX2) expressions in relation to clinical features of patients and elucidate its role in the progression of hepatocellular carcinoma. Methods: Using the Cancer Genome Atlas (TCGA) database, R language was used to analyze the distribution of differentially expressed mRNA in hepatocellular carcinoma. The different expression of CBX2 in HCC and adjacent tissues and its relationship with survival and clinical characteristics of patients were further analyzed. The expression of CBX2 in liver tissues, liver cancer tissue, and L02, HepG2 and SMMC-7721 cell lines was detected by real time-PCR and western blot. The expression of CBX2 was interfered by siRNA in hepatoma cell line. MTT, colony formation, transwell assays, and flow cytometry were used to identify the proliferation, apoptosis, invasion and clone-formation ability of HepG2 and SMMC-7721 cells after CBX2 down-regulation. According to the different data, t-test, ANOVA, chi-square test, and COX regression model were used for statistical analysis. Survival curve was plotted through Kaplan-Meier method. Results: TCGA public database analysis showed that the expression of CBX2 mRNA in hepatocellular carcinoma tissues (7.296 ± 1.6115) was significantly higher than normal liver tissues (4.706 ± 0.940) ( P = 0.000). In addition, the overall survival time of patients with low CBX2 mRNA expression was significantly longer than that of patients with high CBX2 mRNA expression [(5.971 ± 0.411) years vs. (4.650 ± 0.503) years, P = 0.001]. The expression level of CBX2 mRNA was correlated with the pathological TNM stage ( P = 0.025) and differentiation degree ( P < 0.001) of liver cancer. COX regression analysis showed that CBX2 mRNA expression was an independent predictor of patient survival ( P = 0.013). siRNA was transfected and compared with the blank control group. The transgenic ability of HepG2 and SMMC-77221 cells decreased significantly at 72h ( P < 0.05) and 96h ( P < 0.05), and the apoptosis rate (11.430% ± 0.215%) was higher than blank control group (6.6 00% ± 0.170%) ( P = 0.003). The number of invasive cells ((both P < 0.05) and relative colony forming cells ((both P < 0.001) were significantly decreased. In 20 cases of tissue samples, the expression of CBX2 protein (relative expression level 3.020 ± 0.269) in liver cancer was higher than that in adjacent tissues (relative expression level 0.886±0.065) ( P < 0.001). The overall survival time of patients with low CBX2 expression in liver cancer was longer than that of patients with high expression [(3.670 + 0.576) years vs. (0.834 + 0.153) years, P = 0.004]. Conclusion: An evident high expression of CBX2 is an independent poor prognostic factor in hepatoma. Down-regulation of CBX2 expression can inhibit the progression of liver cancer. Therefore, CBX2 may be a prognostic biomarker and a new target for HCC treatment.

Objective: To analyze the clinical and genetic features of immunodeficiency, centromeric instability, and facial anomalies (ICF) syndrome with a case report and literature review. Methods: The clinical data and genetic test of a girl diagnosed with ICF syndrome in the Department of Nephrology and Immunology in Qingdao Women and Children's Hospital in December 2016 were extracted and analyzed. "ICF syndrome" "immunodeficiency, centromeric instability and facial anomalies syndrome" "ICF syndrome and DNMT3B" were used as key words to search Chinese databases and Pubmed for literature until March 2018, and the literature was reviewed. Results: A female patient aged 22 months old with ocular hypertelorism and low-set ears was admitted due to recurrent infection over one year. Laboratory tests showed humoral immune deficiency with IgG<1.34 g/L, IgA<0.060 g/L, and IgM<0.179 g/L, but normal cellular immunity (total T lymphocyte 0.503, hepler T lymphocyte 0.328, cytotoxic T lymphocyte 0.166, natural killer cell 0.184, total B lymphocyte 0.276). Whole-exome sequencing revealed a de novo heterozygous splice site mutation c.922-2A>G in intron 8, and a de novo heterozygous missense mutation c.2477G>A in exon 23 of DNMT3B gene. Chromosome karyotype analysis showed 46, XX, with 64 out of 100 karyotypes showing centromere instability in chromosome 1. Five papers were found which were all in English, with total of 29 patients. Forty-three mutations were reported, including 34 missense, 2 deletion, 1 insertion, 6 splice site mutations. Eleven patients had complex heterozygosis mutations. All patients had centromere instability, humoral immune deficiency and facial dysplasia which were mainly ocular hypertelorism and low-set ears. Most patients had language and motor development delay, and a few were combined with mental retardation. Conclusions: ICF syndrome is a rare autosomal recessive primary immunodeficiency with classic clinical triad manifestations. De novo mutation of DNMT3B gene is one of etiologies according to genetic test.

In the current study, we used heat stress (HS) as an oxidative stress model to examine the effects of hydroxy-selenomethionine (HMSeBA), an organic selenium source, on selenium's bioavailability, antioxidant status, and performance when fed to dairy cows. Eight mid-lactation Holstein dairy cows (141 ± 27 d in milk, 35.3 ± 2.8 kg of milk/d, parity 2 or 3) were individually housed in environmental chambers and randomly assigned to 1 of 2 treatments: inorganic Se supplementation (sodium selenite; SS; 0.3 mg of Se/kg of dry matter; n = 4) or HMSeBA supplementation (0.3 mg of Se/kg of dry matter; n = 4). The trial was divided into 3 continuous periods: a covariate period (9 d), a thermal neutral (TN) period (28 d), and a HS period (9 d). During the covariate and TN periods, all cows were housed in TN conditions (20°C, 55% humidity). During HS, all cows were exposed to cyclical HS conditions (32-36°C, 40% humidity). All cows were fed SS during the covariate period, and dietary treatments were implemented during the TN and HS periods. During HS, cows fed HMSeBA had increased Se concentrations in serum and milk, and total Se milk-to-serum concentration ratio compared with SS controls. Superoxide dismutase activity did not differ between Se sources, but we noted a treatment by day interaction in glutathione peroxidase activity as HS progressively reduced it in SS controls, whereas it was maintained in HMSeBA cows. Supplementation with HMSeBA increased total antioxidant capacity and decreased malondialdehyde, hydrogen peroxide, and nitric oxide serum concentrations compared with SS-fed controls. We found no treatment effects on rectal temperature, respiratory rate, or dry matter intake. Supplementing HMSeBA tended to increase milk yield and decrease milk fat percentage. No other milk composition parameters differed between treatments. We observed no treatment effects detected on blood biochemistry, except for a lower alanine aminotransferase activity in HMSeBA-fed cows. These results demonstrate that HMSeBA supplementation decreases some parameters of HS-induced oxidative stress., (Copyright © 2019 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

Objectives: The difficulty in proliferation and availability and the rapid loss functions of primary human hepatocytes highlight the need to develop an alternative, preferably renewable source of human induced hepatocytes in regenerative medicine. Liver organoids generated on a multiple-cell microenvironment in a 3-dimensional (3D) system can provide a highly efficient solution to this issue., Methods: Human hepatocytes were induced from fibroblasts by the lentiviral expression of FOXA3, HNF1A, and HNF4A. Together with these induced hepatocytes, human umbilical vein endothelial cells and mesenchymal stem cells in a 3D system were used to produce liver organoids. Liver-related gene and protein expression of liver organoids and induced hepatocytes were tested using a 2-dimensional (2D) system., Results: Liver organoids notably increased the expression of hepatic transcription factors, marker genes, transporter genes, and liver metabolism enzyme genes, while it decreased the specific gene expression of fibroblasts. Liver organoids expressed comparable liver-specific proteins, such as ALB, AAT, and HNF4A in the 3D system., Conclusion: Direct reprogramming in multiple-cell microenvironments in 3D systems is more controllable and efficient than cell reprogramming in 2D systems. Liver organoids have the potential for use in disease modeling, pharmaceutical applications, and cellular transplantation., (Copyright © 2018 Elsevier Inc. All rights reserved.)

Objective: To observe the characteristics of the interstitial fluid (ISF) drainage in the Alzheimer's disease (AD) rats through magnetic resonance imaging (MRI) tracer gadolinium-diethylene triamine pentacetic acid (Gd-DTPA)spread in the brain extracellular space (ECS) and to discuss the role of aquaporin-4 (Aqp4) in the AD., Methods: Wild type SD rats (300-350 g) and Aqp4 gene knock out (Aqp4 -/- ) SD rats (300-350g) were divided into Sham group, AD group, Aqp4 -/- -Sham group and Aqp4 -/- -AD group. Sham group and Aqp4 -/- -Sham group were injected with saline by intraperitoneal each day for 6 weeks, and the AD group and Aqp4 -/- -AD group were injected with D-galactose by intraperitoneal each day for 6 weeks. MRI tracer Gd-DTPA (10 mmol/L, 2 μL) was injected into the hippocampus of the rats. MRI scan was performed at the end of 0.5 h, 1.5 h, 1 h, 2 h, and 3 h to observe the dynamic distribution of the Gd-DTPA in the hippocampus and the diffusion rate D * , clearance rate k' and half-life t 1/2 measured., Results: The diffusion rate D* in Sham group was (2.66±0.36)×10 -6 mm2/s, the diffusion rate D * in AD group was (2.72±0.62)×10-6 mm 2 /s, the diffusion rate D * in Aqp4 -/- -Sham group was (2.75±0.47)×10 -6 mm 2 /s, the diffusion rate D * in Aqp4 -/- -AD group was (2.802±0.55)×10 -6 mm 2 /s, and there was no statistically significant difference in the four groups (One-Way ANOVA, P>0.05).The clearance rate k' in Sham group was (4.57±0.14)×10 -4 /s, the clearance rate k' in AD group was (3.68±0.22)×10 -4 /s, the clearance rate k' in Aqp4 -/- -Sham group was (3.17±0.16)×10 -4 /s, the clearance rate k' in Aqp4 -/- -AD group was (2.59±0.19)×10 -4 /s, and there was significant difference in the four groups (One-Way ANOVA, P<0.05). The half-life t 1/2 in Sham group was (0.67±0.12) h, the half-life t 1/2 in AD group was (0.88±0.08) h, the half-life t 1/2 in Aqp4 -/- -Sham group was (1.12±0.15) h, the half-life t 1/2 in Aqp4 -/- -AD group was (1.58±0.11) h, and there was significance difference in the four groups(one-way ANOVA,P<0.05)., Conclusion: The ISF drainage is slow after AD and the loss of Aqp4 in the AD makes the ISF drainage obviously slow down, Aqp4 plays an important role in AD to remove the metabolism of waste out of the brain.

The tissue kinetics of cyadox, an antibacterial agent used in food animals, and its major metabolites in pigs, chickens, and carp were investigated followed by a complete dietary exposure assessment to evaluate the food safety of cyadox. Cyadox and its major metabolites, bisdeoxycyadox (Cy1), 4-desoxycyadox (Cy2), N-(quinoxaline-2-methyl)-cyanide acetyl hydrazine (Cy4), quinoxaline-2-carboxylic acid (Cy6), and 2-hydromethyl-3-hydroxy-quinoxaline (Cy12), were simultaneously quantitated with a high-performance liquid chromatography-ultraviolet (HPLC-UV) method. Pigs, chickens, and carp were fed with 150 mg/kg cyadox in feed for consecutive 60, 40, and 30 days, respectively. The residue amount of cyadox and its major metabolites in liver, kidney, muscle, and fat (skin) tissues was determined. Cy2 was below the limit of quantitation even at the withdrawal time of 6 hr, cyadox, Cy4, Cy6, and Cy12 could be detected at 6-24 hr with low level less than 50 μg/kg. By contrast, Cy1 persisted for 3 days in the kidney of pigs and chickens, and in the liver of carp. Based on these residue depletion data and previous toxicology results, the global estimated chronic dietary exposure assessment of cyadox for general population was conducted, indicating a zero withdrawal time (WDT) may be appropriate for cyadox in food animals when used in feed for prolonged administration. These results provide analytical techniques and safety standards suitable for residue monitoring of cyadox in food animals., (© 2017 John Wiley & Sons Ltd.)

PHACE syndrome is a syndrome of multiple organ and multisystem abnormalities associated with infantile segmental hemangioma, characterized by abnormal posterior fossa development, infant hemangioma, aortic abnormalities, aortic coarctation and heart defects, eye anomalies and other symptoms. The incidence of the disease is low, but there exist life-threatening factors. Once clinically diagnosed, it should be highly valued and multidisciplinary consultation must be conducted. This article reviews the diagnostic criteria of PHACE syndrome and its associated facial segmental hemangioma, as well as the treatment and prognosis of brain abnormalities.

Objective: To evaluate the effect of breast massage at different time in the early period on maternal lactation after cesarean section. Methods: 80 women delivered by cesarean section were randomly selected from maternity ward of a hospital in Shandong province during Jan. 2013 to Jan. 2015; which were divided into four groups, with 20 patients in each. Three groups received 3 times of breast massage every 24 hoursbeginning from 2, 12 and 24 h after cesarean section, respectively. The control group didn't receive any breast massage. The starting time and status of lactation were observed and recorded after cesarean section. 5 ml venous blood sample was drawn from each patient respectively at 2 h before cesarean, 6, 12, 24, 48 and 72 h after cesarean to test the level of serum prolactin. The lactation status of each group was compared. Results: The P (50) ( P (25)- P (75)) of starting time of lactation of the three massage groups and control group were 3 (2-6) h, 4 (2-8) h, 4 (3-12) h and 4 (2-12) h, respectively, whose differences showed no statistical significance ( H =3.32, P= 0.345).The number of delivered women with adequate lactation 24 hours after cesarean was 10 in the group who received massage beginning from 2 h after cesarean; while the number was only 2 in the control group. The number of delivered women with adequate lactation 48 hours after cesarean was 18 in the group who received massage beginning from 2 h after cesarean; while the number was 8 in the control group. The differences showed statistical significances ( P values were 0.021 and 0.008, respectively). The serum prolactin level in the group of delivered women who received massage from 2 h after cesarean was separately (195.9±78.5), (176.0±96.5), (216.4±110.0), (190.0±56.8) and (184.8±69.6) μg/L at 2, 12, 24, 48 and 72 h after cesarean, which were significantly higher than those in the control group (which were (128.8±40.6), (127.3±66.8), (162.2±58.8), (145.1±64.7) and (141.7±49.3) μg/L, respectively) ( P= 0.007). Conclusion: Breast massage beginning from 2 hours after cesarean section can effectively improve the lactation status of delivered women.

The aim of this paper is to study the correlation between the cardiac output index value and the prognosis of patients with heart disease with the help of Pulse-induced Contour Cardiac Output (PICCO) and computed tomography (CT). Three hundred eighteen patients with pathologically confirmed heart disease were selected as study subjects. The Sime ns Av anto 1.6T type superconducting magnetic resonance meter and GE Light Speed Pro 48 slice spiral CT were used for the phased array coil scanning and enhanced scanning, respectively and to analyze the relationship between the cardiac output index value and prognosis in patients with heart disease with the help of PICCO and CT. The accuracy rates of PICCO and CT in determining the cardiac output index of heart disease were 93.2% and 76.1%, respectively, which were significantly different (P less than 0.05). The coincidence rate between the results of the cardiac output index and pathological results of heart disease from PICCO and CT were 57.6%and 54.7%, respectively. According to the χ2 test results, the diagnostic results of PICCO were consistent with the pathological results of the heart disease group (Kappa = 0.039, 0.496, P less than 0.05). The coincidence rate for the five-year survival rate of PICCO/CT with the total survival rate and the results of the patients were 56.7% and 43.8%, respectively. The coincidence rate of PICCO and CT diagnosis results with the cardiac output index values, as well as prognosis results, was high. In judging the value of the cardiac output index and the prognosis of patients with heart disease, PICCO was superior to CT. The diagnostic results of PICCO have a strong correlation with the pathological results, therefore, PICCO can be used to accurately determine the cardiac output index and the prognoses of patients with heart disease.

Rectal cancer is a commonly observed tumor in clinics, and epithelial-mesenchymal transition (EMT) is very important for tumor invasion and metastasis. We established a rectal cancer HCT-116 cell hypoxia model and detected cell proliferation, invasion, and EMT-related protein expression in this model, aiming to analyze the effect of hypoxia on rectal cancer cell EMT. Rectal cancer cell line HCT-116 was cultured in normoxic, hypoxic, or anaerobic environment, and hypoxia-inducible factor-1α (HIF-1α) mRNA expression was detected in the cells by real-time PCR. Cell proliferation was tested by MTT assay; cell invasion was determined by transwell assay, and HIF-1α, epithelial-cadherin, and Snail protein levels were evaluated by western blot analysis. HIF-1α mRNA level significantly increased in the anaerobic group compared to that in the normoxic and hypoxic groups (P < 0.05). HCT-116 cell proliferation in the anaerobic group was obviously higher than that in the other two groups, with the hypoxic group showing stronger proliferative ability than the normoxic group (P < 0.05). Compared to the normoxic group, the HCT-116 cells demonstrated enhanced cell invasion and migration in hypoxic and anaerobic groups. HIF-1α and Snail expressions were upregulated, whereas epithelial-cadherin expression had declined in the hypoxic and anaerobic groups, compared to those in the normal control (P < 0.05). Therefore, hypoxia promoted rectal cancer cell progress by increasing HIF-1α to induce EMT.

Objective: To investigate the contents and features of drug-induced liver injury (DILI) database called LiverTox, as well as 37 herbal preparations included in this database. Methods: Firstly, the source and contents of LiverTox were briefly introduced, including the clinical features, types, severity, and causality assessment scale of DILI. Secondly, detailed information of 37 herbal preparations included in the class of "Herbals and Dietary Supplements" were extracted, including drug name, origin, efficacy, constituents, type of liver injury, and manifestations, to perform a preliminary statistical analysis. Finally, a comparative analysis was performed between such information and current knowledge of Chinese herbal medicine-induced liver injury in China. Results: LiverTox was a DILI database with open access and rich information and provided practical information on treatment, typing, causality assessment, and treatment. Among the 37 herbal preparations, 28 had the risk of liver injury. The most common indication was weight loss, followed by arthritis and constipation. The latency of hepatotoxicity ranged from 4 weeks to 6 months. Compared with the current knowledge in China, there were differences in the varieties and indications for herbal preparations with hepatotoxicity included in LiverTox, and many herbals with acknowledged hepatotoxicity in China were not included. Conclusion: LiverTox database is concise and practical, but there are certain differences between the herbal preparations included in this database and current knowledge in China.

Breast cancer is among the most common causes of cancer-related death in women worldwide. Previous studies have demonstrated an association between prolonged estrogen exposure and increased risk of breast cancer. Uridine 5'-diphospho-glucuronosyltransferase 1-1 (UGT1A1) plays a significant role in the detoxification of estrogens. Two major genetic polymorphisms have been identified in the UGT1A1 locus. UGT1A1*28 has been previously linked to increased risk of breast cancer. The aim of this study was to elucidate the possible correlation between UGT1A1*6, a single nucleotide polymorphism causing a Gly71Arg substitution, and breast cancer susceptibility. Forty-six women diagnosed with breast cancer, 15 patients with gastrointestinal cancer, and 13 healthy women were recruited to this study. The genotype in the polymorphic UGT1A1 locus was determined by DNA sequencing. The frequency of each genotype was compared among the three groups. The frequency of the UGT1A1*6 allele was significantly higher in breast cancer and gastrointestinal cancer patients than that in healthy females (both P < 0.05). No significant associations were observed between the UGT1A1*6 polymorphism and estrogen receptor, progesterone receptor, HER-2 expression status, menstrual status, or metastasis (all P > 0.05). Therefore, the UGT1A1*6 polymorphism was deduced to be a risk factor for breast cancer in women of Han Chinese ethnicity. UGT1A1 may serve as a therapeutic target for the prevention and treatment of breast cancer and other estrogen-related diseases., Competing Interests: The authors declare no conflict of interest.

The objective of the present study was to investigate whether hepatic fibrosis difference of supersonic shear imaging (SSI) value existed in patients with different etiological chronic hepatic diseases. Retrospective analysis was used to study chronic hepatitis. All the subjects were diagnosed by shear wave elastography and percutaneous liver biopsy. The shear moduli were analyzed to check whether any difference existed between groups. For the chronic hepatitis B, autoimmune hepatitis and fatty hepatitis, the shear moduli in S0 stage were (8.50±3.1)kPa, (9.41±2.5)kPa, (8.97±3.8)kPa; the shear moduli in S1 stage were (9.54±3.0)kPa, (10.42±5.1)kPa, (9.51±4.6)kPa; the shear moduli in S2 stage were (11.77±4.8) kPa, (13.25±5.6)kPa, (11.03±6.0)kPa; the shear moduli in S3 stage were (14.96±6.1)kPa, (19.03±7.8) kPa, (15.38±7.8)kPa; the shear moduli in S4 stage were (20.36±7.5)kPa, (24.99±9.5)kPa, (19.53±5.6)kPa. Shear wave elastography could measure the different etiological chronic hepatic diseases.

Phenol, also known as carbolic acid or phenic acid, is a priority pollutant in aquatic ecosystems. The present study has investigated metabolic activities and transcription profiles of cytochrome P450 enzymes in Chironomus kiinensis under phenol stress. Exposure of C. kiinensis larvae to three sublethal doses of phenol (1, 10 and 100 µM) inhibited cytochrome P450 enzyme activity during the 96 h exposure period. The P450 activity measured after the 24 h exposure to phenol stress could be used to assess the level (low or high) of phenol contamination in the environment. To investigate the potential of cytochrome P450 genes as molecular biomarkers to monitor phenol contamination, the cDNA of ten CYP6 genes from the transcriptome of C. kiinensis were identified and sequenced. The open reading frames of the CYP6 genes ranged from 1266 to 1587 bp, encoding deduced polypeptides composed of between 421 and 528 amino acids, with predicted molecular masses from 49.01 to 61.94 kDa and isoelectric points (PI) from 6.01 to 8.89. Among the CYP6 genes, the mRNA expression levels of the CYP6EW3, CYP6EV9, CYP6FV1 and CYP6FV2 genes significantly altered in response to phenol exposure; therefore, these genes could potentially serve as biomarkers in the environment. This study shows that P450 activity combined with one or multiple CYP6 genes could be used to monitor phenol pollution.

This study aims to investigate mechanism of YiQi ChuTan Recipe (YCR) for inhibiting epithelial mesenchymal transition (EMT) of A549 cells under hypoxia. Flow cytometry was used to optimize YCR dosage by measuring A549 apoptosis, which were subjected to different treatments, including normal condition, hypoxia, hypoxia+YCR. Cell morphology and expression of EMT were measured with differential interference contrast microscopy, real-time PCR and western blot. Optimized condition of 4 mg/ml YCR and 2% O2 for 72 h was used to establish hypoxia. Under hypoxic condition, morphology of A549 cells changed from oblate fusi-form to elongated spindle. E-cadherin expression decreased while vimentin and fibronectin increased. EMT-related genes expression were significantly increased in hypoxia group compared to control group (P<0.05). After treatment with YCR, mesenchymal cells obviously decreased and EMT-related genes expression was significantly decreased (P<0.05). Changes of E-cadherin, vimentin and fibronection were significantly attenuated by YCR when compared to hypoxia group. Expression of proteins GRP78, SRC, MAPK, smad2/3 were significantly increased in hypoxia group compared to control group, but was significantly inhibited by YCR treatment. In conclusion, A549 cells underwent EMT under hypoxia while YCR reversed the EMT through GRP78, smad2/3 and SRC/MAPK signal pathway.

4-Coumarate: coenzyme A ligase (4CL) catalyzes the conversion of hydroxycinnamates into corresponding CoA esters for biosynthesis of flavonoids and lignin. It has been widely studied in seed plants; however, it is poorly characterized in ferns. In this study, we identified 4CLs genes in ferns D. fragrans (L.) Schott (Df4CL gene) by rapid amplification of cDNA ends (RACE), and then investigated the expressions of the genes by real-time PCR, and determined total flavonoids and lignin contents. The results showed that four members of the 4CL genes were found from this species, which named Df4CL 1, 2, 3, and 4 genes. Their full-length cDNA was sequenced. Also, our analyses showed that the amino acid sequences derived from these cDNAs exhibited similar conserved regions (Box I and Box II), and substrate-binding regions compared to 4CLs isolated from seed plants.At the same time, the developmental and stress-induced gene expression patterns showed that the changes on the expression levels of Df4CL genes affected the levels of flavonoids and lignin. In conclusion, we identified 4CLs genes in ferns D. fragrans and analyzed the expressions of these genes, and finally explored the relationship between the expressions of 4CLs and syntheses of flavonoids and lignin.