Objective: To investigate the regulatory effect of Jieduan Niwan Formula (JDNWF) drug-containing serum on AMPK-mediated mitochondrial quality control in D-GalN-induced HepG2 cells. Methods: Twenty male Wistar rats were randomly divided into blank control and JDNWF-containing serum groups, 10 rats per group. The JDNWF-containing serum group was gavaged with JDNWF (21.7 g/kg), whereas the blank control group was gavaged with saline. Blood was collected to prepare JDNWF-containing and blank control serum. Cell viability, mitochondrial damage indicators, and MQC pathway protein expression levels were evaluated to determine the optimal volume fraction of JDNWF. HepG2 cells were divided into control, D-GalN, DMSO, AMPK inhibitor, JDNWF drug-containing serum, and JDNWF drug-containing serum plus AMPK inhibitor groups, and corresponding drug interventions were administered to each group. Cells were collected after the interventions, and the CCK-8 assay was used to measure cell viability, the 2′-7′-dichlorodihydrofluorescein diacetate fluorescent probe was used to detect reactive oxygen species (ROS) levels, JC-1 was used to detect mitochondrial membrane potential, thiobarbituric acid was used to measure malondialdehyde (MDA) levels, WST-8 was used to measure superoxide dismutase (SOD) activity, and western blotting was used to detect the expression levels of mitochondrial quality control-related proteins, including p-AMPK, AMPK, PGC-1α, NRF1, TFAM, MFN2, and DRP1. Results: 5% JDNWF drug-containing serum most significantly restored cell viability, mitochondrial damage markers, and MQC pathway protein expression in the model group. Therefore, it was chosen for intervention in subsequent experiments. Compared to the control group, the cell viability of the D-GalN, DMSO, and AMPK inhibitor groups was significantly reduced (P<0.01). In contrast, the heterogeneity of mitochondrial membrane potential, ROS, and MDA levels was significantly increased (P<0.01), and SOD activity was significantly decreased (P<0.01). The p-AMPK, PGC-1α, NRF1, TFAM, MFN2, and DRP1 protein expression levels were significantly decreased (P<0.01). After JDNWF drug-containing serum intervention, compared to the DMSO group, cell viability significantly increased (P<0.01), mitochondrial membrane potential heterogeneity, ROS, and MDA levels significantly decreased (P<0.01), SOD activity significantly increased (P<0.01), and p-AMPK, PGC-1α, NRF1, TFAM, and MFN2 protein expression levels significantly increased (P<0.01), whereas DRP1 protein expression significantly decreased (P<0.01). Compared to the JDNWF drug-containing serum group, the cell viability in the JDNWF plus AMPK inhibitor group significantly decreased (P<0.01), mitochondrial membrane potential heterogeneity and ROS levels significantly increased (P<0.01), MDA levels significantly increased (P<0.05), SOD activity significantly decreased (P<0.05), p-AMPK, PGC-1α, NRF1, and TFAM protein expression levels significantly decreased (P<0.01), MFN2 protein expression significantly decreased (P<0.05), and DRP1 protein expression significantly increased (P<0.01). Conclusion JDNWF drug-containing serum may restore mitochondrial function and improve D-GalN-induced HepG2 cell injury by regulating AMPK-mediated mitochondrial quality control.

The use of nitrification inhibitors has been suggested as a strategy to decrease cadmium(Cd)accumulation in crops.However,the most efficient nitrification inhibitor for mitigating crop Cd accumulation remains to be elucidated,and whether and how changes in soil microbial structure are involved in this process also remains unclear.To address these questions,this study applied three commercial nitrification inhibitors,namely,dicyandiamide(DCD),3,4-dimethylpyrazole phosphate(DMPP),and nitrapyrin(NP),to pakchoi.The results showed that both DCD and DMPP(but not NP)could efficiently decrease Cd concentrations in pakchoi in urea-and ammonium-fertilized soils.In addition,among the three tested nitrification inhibitors,DMPP was the most efficient in decreasing the Cd concentration in pakchoi.The nitrification inhibitors decreased pakchoi Cd concentrations by suppressing acidification-induced Cd availability and reshaping the soil microbial structure;the most effective nitrification inhibitor was DMPP.Ammonia oxidation generates the most protons during nitrification and is inhibited by nitrification inhibitors.Changes in environmental factors and predatory bacterial abundance caused by the nitrification inhibitors changed the soil microbial structure and increased the potential participants in plant Cd accumulation.In summary,our study identified DMPP as the most efficient nitrification inhibitor for mitigating crop Cd contamination and observed that the soil microbial structural changes caused by the nitrification inhibitors contributed to decreasing Cd concentration in pakchoi.

BACKGROUND: 3D-printing is widely used in regenerative medicine and is expected to achieve vaginal morphological restoration and true functional reconstruction. Mesenchymal stem cells-derived exosomes (MSCs-Exos) were applyed in the regeneration of various tissues. The current study aimed to explore the effctive of MSCs-Exos in vaginal reconstruction. METHODS: In this work, hydrogel was designed using decellularized extracellular matrix (dECM) and gelatin methacrylate (GelMA) and silk fibroin (SF). The biological scaffolds were constructed using desktop-stereolithography.The physicochemical properties of the hydrogels were evaluated; Some experiments have been conducted to evaluate exosomes’ effect of promotion vaginal reconstruction and to explore the mechanism in this process. RESULTS: It was observed that the sustained release property of exosomes in the hydrogel both in vitro and in vitro.The results revealed that 3D scaffold encapsulating exosomes expressed significant effects on the vascularization and musule regeneration of the regenerative vagina tissue. Also, MSCs-Exos strongly promoted vascularization in the vaginal reconstruction of rats, which may through the PI3K/AKT signaling pathway. CONCLUSION The use of exosome-hydrogel composites improved the epithelial regeneration of vaginal tissue, increased angiogenesis, and promoted smooth muscle tissue regeneration. 3D-printed, lumenal scaffold encapsulating exosomes might be used as a cell-free alternative treatment strategy for vaginal reconstruction.

Objective:To explore the value of proton density fat fraction(PDFF) based on histogram analysis for quantification hepatic steatosis and fibrosis in rabbit model and the interference of hepatic fibrosis to the evaluation of hepatic steatosis with PDFF.Methods:From March to November 2020, 135 New Zealand white rabbits were randomly divided into control group ( n=30) and experimental group ( n=105) using a random number table. The volume ratio of CCl 4 and olive oil was 1∶1 to prepare 50% CCl 4 oil solution, and experimental rabbits were subcutaneously injected with the oil solution. An equal dose of normal saline was subcutaneously injected for control group rabbits. At the end of the 4 th, 8 th, and 12 th week, 35 in the experimental group and 10 rabbits in the control group were randomly selected to conduct the mDixon-Quant scanning, and histogram analysis of PDFF was analyzed including volume, mean, median, standard deviation, 25 th, 50 th, 75 th, 90 th quantile, skewness, kurtosis, entropy and inhomogeneity. After the examination, the rabbits were sacrificed and the liver percentage of steatosis (PSH) and fibrosis (POF) were recorded by semi-quantitative analysis. Spearman correlation analysis was used to correlate PDFF with PSH and POF. Multiple linear regression analysis was used to determine independent PDFF histogram parameters for evaluating PSH and POF. A receiver operator characteristic (ROC) curve was used to assess the diagnostic accuracy of PDFF for discriminating mild from moderate-severe hepatic steatosis and mild from moderate-severe hepatic fibrosis with median of PSH or POF for dichotomy, and DeLong test was used to compare the area under the curve (AUC). With the correction of hepatic fibrosis, correlation coefficient and AUC were compared of PDFF for discrimination mild from moderate-severe hepatic steatosis. Results:The PDFF mean, median, standard deviation, 75 th, 90 th showed correlation with PSH ( r=0.558, 0.522, 0.319, 0.723, 0.646, -0.589, all P<0.05). The entropy and 75 th were independent parameters for evaluating PSH (β=2.347, -5.960, P=0.018, 0.001). The PDFF 75 th was the optimal parameter for discriminating mild from moderate-severe hepatic steatosis with AUC=0.915 ( P=0.001). The PDFF volume, mean, median, standard deviation, 75 th, 90 th, entropy showed correlation with POF ( r=0.355, 0.393, 0.376, 0.298, 0.485, 0.426, -0.681, all P<0.05). The entropy, standard deviation and volume (β=-11.041, 1.356, 0.190, P=0.001, 0.026, 0.016) were independent parameters for evaluation of hepatic fibrosis, and the entropy was the optimal parameter for hepatic fibrosis (AUC=0.771, P=0.001). The correlation between PSH and PDFF 75 th was less pronounced when fibrosis was present ( r=0.512, P=0.001) than when fibrosis was absent ( r=0.751, P=0.002). The PDFF 75 th showed a significant difference in discriminating mild hepatic steatosis from moderate-severe hepatic steatosis after correction of POF (AUC=0.895, 0.950, Z=2.970, P=0.025). Conclusions:PDFF based on histogram analysis provided a noninvasive, accurate estimation of quantification for hepatic steatosis and fibrosis. Hepatic fibrosis reduced the correlation between hepatic steatosis and PDFF and the presence of hepatic fibrosis can confound the quantification of hepatic steatosis with PDFF.

Objective: To investigate the prevalence of adolescents dietary behavior in Shanghai, and to explore emotional influence on dietary behavior. Methods: A total of 7 456 students from 10 junior and 6 senior high schools in Shanghai were selected to participate in the questionnaire survey with the stratified random cluster sampling method. The survey included general information, eating behavior, PHQ-2 and GAD-7. Results: During the past week, the proportion of adolescents in Shanghai reported consumption of sugar sweetened beverages, sweet desserts, frequent fried food and fast food (≥4 times/week) were 13.26%, 16.90%, 6.99 % and 13.01%, respectively. The proportion of students reported consumption of fruits, vegetables, milk and breakfast every day were 56.96%, 73.00%, 65.03% and 76.11%, respectively. There were significant differences by sex and educational stages(both P <0.05). Adolescents with depression or anxiety have a higher incidence of unhealthy eating behaviors than those without depression or anxiety( P <0.01). After adjusting for gender, school, accommodation, grades, pocket money and social class, depression and anxiety increase the risk of various unhealthy eating behaviors in adolescents( P <0.05). Compared with those without anxiety, the risks of sugar sweetened beverages consumption (≥1 time/d) among adolescents with mild and moderate to severe anxiety were 1.42 times (95% CI =1.20-1.67) and 2.51 times (95% CI =2.09-3.01), the risks of insufficient fruits consumption (<1 time/d) were 1.30 times (95% CI =1.16-1.45) and 1.28 times (95% CI =1.11-1.47), the risks of insufficient vegetable consumption (<1 time/d) were 1.35 times (95% CI =1.20-1.52) and 1.41 times(95% CI =1.21-1.65), the risks of insufficient milk consumption (<1 time/d) were 1.29 times (95% CI =1.15-1.44) and 1.20 times(95% CI =1.04-1.39), and the risks of breakfast skipping were 1.75 times (95% CI =1.54-1.99) and 2.97 times (95% CI =2.55-3.46) among adolescents with mild and moderate to severe anxiety. Conclusion The proportion of unhealthy eating behaviors among adolescents in Shanghai is still high. Early education and intervention for students eating behaviors should be carried out, and attention should be paid to the occurrence of adolescents negative emotions, so as to reduce the risk of unhealthy eating behaviors among adolescents through the promotion of mental health.

The importance of structural variants(SVs)for human phenotypes and diseases is now recognized.Although a variety of SV detection platforms and strategies that vary in sensitivity and specificity have been developed,few benchmarking procedures are available to confidently assess their performances in biological and clinical research.To facilitate the validation and application of these SV detection approaches,we established an Asian reference material by characterizing the genome of an Epstein-Barr virus(EBV)-immortalized B lymphocyte line along with identified benchmark regions and high-confidence SV calls.We established a high-confidence SV callset with 8938 SVs by integrating four alignment-based SV callers,including 109x Pacific Biosciences(PacBio)continuous long reads(CLRs),22 x PacBio circular consensus sequencing(CCS)reads,104x Oxford Nanopore Technologies(ONT)long reads,and 114×Bionano optical mapping plat-form,and one de novo assembly-based SV caller using CCS reads.A total of 544 randomly selected SVs were validated by PCR amplification and Sanger sequencing,demonstrating the robustness of our SV calls.Combining trio-binning-based haplotype assemblies,we established an SV benchmark for identifying false negatives and false positives by constructing the continuous high-confidence regions(CHCRs),which covered 1.46 gigabase pairs(Gb)and 6882 SVs supported by at least one diploid haplotype assembly.Establishing high-confidence SV calls for a benchmark sample that has been characterized by multiple technologies provides a valuable resource for investigating SVs in human biology,disease,and clinical research.

Objective:To explore the relationship between rs2010963, rs3025039 and rs699947 gene polymorphism of vascular endothelial growth factor(VEGF) gene and bronchopulmonary dysplasia(BPD) in Mongolian premature infants.Methods:A case-control design was used to collect 50 cases of Mongolian premature infants who were hospitalized at the Affiliated Hospital of Inner Mongolia Medical University and diagnosed with BPD from January 2016 to December 2020 as the observation group, while 56 cases of non-BPD premature infants of the same nationality and time period were selected as the control group.Using PCR method to detect the genotype and allele distribution of the VEGF gene rs2010963, rs3025039 and rs699947 locus.Combining clinical data to analyze whether the above gene loci were related to the onset of premature infants with Mongolian BPD in our area.Results:Through genetic testing, it was found that CC, CA and AA genotypes can be detected at the rs699947 site of VEGF gene in premature infants in both the observation group and the control group.The frequencies of the three genotypes in the observation group were 16.0%, 24.0%, and 60.0%, respectively; the frequency of the C allele was 28.0%, the frequency of the A allele was 72.0%, and the frequency of the three genotypes in the control group was 32.1.%, 32.1% and 35.7%, respectively.The frequency of C allele was 48.2%, the frequency of G allele was 51.8%, and the allele and genotype frequencies of this locus between the observation group and the control group were significant differences from those of the control group( P<0.05). Conclusion:The polymorphism of VEGF gene rs699947 locus is associated with the occurrence and development of BPD in Mongolian premature infants, and allele A may be a susceptible factor.

Objective: To learn the sequence characteristics of C2-V4 of HIV-1 env genes and the epitope variation of representative broadly monoclonal neutralizing antibody in Fuxin, so as to provide evidence for the HIV-1 variation trend and the biological characteristics of V3 loop. Methods : The whole blood samples of 112 HIV-1 cases in Fuxin Health Service Center from 2018 to 2019 were collected and the DNA was extracted. The C2-V4 of env genes were amplified by nested-PCR and the PCR products were subjected to sequencing. The bioinformatics analysis was carried out using MEGA software, and the V3-tip motifs, co-receptors, net charge and characteristic amino acids were analyzed using HIV Database. Results: Totally 101 effective gene sequences were obtained, and 5 types of V3-tip motifs were found. Among them, 77 pieces of GPGQ ( 76.24% ) were found in CRF01_AE, CRF07_BC, CRF65_cpx and G subtypes; 19 pieces of GPGR ( 18.81% ) were found in CRF01_AE, CRF07_BC and B subtypes; 3 pieces of GPGH, 1 piece of GPGK and 1 piece of GPGA were only found in CRF01_ AE subtype. The co-receptor was mainly CCR5 ( 84, 83.17% ) . The net charge numbers of V3 loops in CRF01_ AE, CRF07_ BC, B, CRF65_cpx and G were 3.28±1.17, 3.22±0.92, 4.25±0.83, 2.50±0.50 and 3, respectively. The mutation rates of neutralizing antibodies binding b12 and VRC01 were 0-9.90%. The deletion rates of N-glycosylation sites of 295 and 332 were 18.81% and 14.85%, without the loss of both sites. Conclusions The HIV-1 strains in Fuxin from 2018 to 2019 are macrophage-tropic and non-syncytium-inducing, with GPGQ as the main type of V3-tip motif, CCR5 as the main co-receptor, slow replication and low ability to escape neutralizing antibodies.

OBJECTIVE To evaluate the embryo toxicity of Shuanghuanglian (SHL) by the combination of a human placental barrier model and embryonic stem (ES) cell test model.METHODS A human placental barrier model was set up by placenta slice culture and Ussing chamber.SHL 0.2,0.4,0.8,1.6,3.2,6.4 and 12.8 g· L-1 was added into the maternal side of the human placental model,respectively.All the media was collected respectively from the matemal side and fetal side 60 min later and taken as the SHL containing medium.ES cells (D3 line) and embryonic fibroblast cells (BALB/c 3T3) were cultured with the SHL containing medium respectively from the maternal side and the fetal side for 10 d.Cell viability was detected by MTT assay,and 50％ survival inhibitory ratio of ES and 3T3 cells by SHL was calculated.ES cells were incubated with the SHL containing medium from the matemal side or fetal side when they differentiated to cardiac myoblasts using hanging drop-suspension-attachment method.Messenger RNA of myosin heavy chain genes (β-MHC) was detected by Q-PCR for differentiation ratio,and 50％ differentiation inhibitory ratio of ES cells by SHL was calculated.A statistics formula was used for prediction of SHL embryotoxicity potential.RESULTS The IC50 of SHL in the matemal side of the human placental model for 3T3 cell survival,ES cell survival and ES differentiation was 1.97,0.84 and 0.48 g· L-1,respectively.According to the criteria for embryo toxicity evaluation,SHL had weak embryo toxicity.However,the IC50 of SHL in the fetal side of the human placental model for 3T3 cell survival,ES cell survival and ES differentiation was 3.19,2.57 and 0.95 g· L-1,respectively.According to the criteria for embryo toxicity evaluation,the supernatant containing SHL that went through the placental barrier had no embryo toxicity.CONCLUSION SHL is safe in the test concentration range during pregnancy.It is more scientific to evaluate embryo toxicity of drugs by ES cell test with the samples obtained through the placental barrier during pregnancy.

Objective: To investigate hepatitis C virus(HCV)genotyping and the serum HCV-RNA concentration in patients infected with different HCV genotypes and to provide information for evaluation of disease condition and anti-viral treatment efficacy. Methods: A total of 60 anti-HCV positive serum samples were collected before antiviral treatment. RT-PCR was performed for the 5′ non-cording region and was followed by nucleotide sequencing for HCV genotyping. Meanwhile, serum HCV-RNA concentration was detected by quantitative PCR. SPSS21.0 and Graphpad Prism 5.0 software were used for data analysis. Analysis of variance (ANOVA) was used for comparison among multi-groups and the t-test was used for comparison between two groups. Results: The frequencies of HCV genotypes 1b, 3a, 1a and 2a were 48.3% (29/60), 23.3% (14/60), 16.7% (10/60) and 10% (6/60), respectively. And, there is one subtype 2c was detected in this study. The mean serum viral concentration with standard deviation of HCV in genotype 1a, 1b, 2a, and 3 a were 5.46±1.19, 6.22±0.78, 5.47±0.65, and 5.38±0.98 log₁₀ (IU/ml) respectively. Conclusions The infection rate of HCV genotype 1 was significantly higher than that of genotype 2 and 3 (P<0.01). The statistical analysis showed the serum HCV-RNA concentration in patients with subtype 1b was significantly higher than that of the subtype group 1 a, 2 a, and 3 a (P<0.05). The study of the relationship between HCV genotypes and the serum HCV-RNA concentration may contribute to anti-viral treatment prescription for hepatitis C patients.