Liver regeneration following injury aids the restoration of liver mass and the recovery of liver function. In the present study we investigated the contribution of megakaryocytic leukemia 1 (MKL1), a transcriptional modulator, to liver regeneration. We report that both MKL1 expression and its nuclear translocation correlated with hepatocyte proliferation in cell and animal models of liver regeneration and in liver failure patients. Mice with MKL1 deletion exhibited defective regenerative response in the liver. Transcriptomic analysis revealed that MKL1 interacted with E2F1 to program pro-regenerative transcription. MAPKAPK2 mediated phosphorylation primed MKL1 for its interaction with E2F1. Of interest, phospholipase d2 promoted MKL1 nuclear accumulation and liver regeneration by catalyzing production of phosphatidic acid (PA). PA administration stimulated hepatocyte proliferation and enhanced survival in a MKL1-dependent manner in a pre-clinical model of liver failure. Finally, PA levels was detected to be positively correlated with expression of pro-regenerative genes and inversely correlated with liver injury in liver failure patients. In conclusion, our data reveal a novel mechanism whereby MKL1 contributes to liver regeneration. Screening for small-molecule compounds boosting MKL1 activity may be considered as a reasonable approach to treat acute liver failure.

A series of new monobactam sulfonates is continuously synthesized and evaluated for their antimicrobial efficacies against Gram-negative bacteria. Compound 33a (IMBZ18G) is highly effective in vitro and in vivo against clinically intractable multi-drug-resistant (MDR) Gram-negative strains, with a highly druglike nature. The checkerboard assay reveals its significant synergistic effect with β-lactamase inhibitor avibactam, and the MIC values against MDR enterobacteria were reduced up to 4-512 folds. X-ray co-crystal and chemoproteomic assays indicate that the anti-MDR bacteria effect of 33a results from the dual inhibition of the common PBP3 and some class A and C β-lactamases. Accordingly, preclinical studies of 33a alone and 33a‒avibactam combination as potential innovative candidates are actively going on, in the treatment of β-lactamase-producing MDR Gram-negative bacterial infections.

Objective:To study the effects of gender on clinical outcomes of extremely low birth weight infants (ELBWI) and to analyze the risk factors of mortality.Methods:From January 2011 to December 2020, ELBWI (birth weight <1 000 g) admitted to the Neonatology Department of our hospital were retrospectively studied. The infants were assigned into the male group and the female group. Incidences of major complications, survival rate and mortality rate were compared between the two groups. The infants were also assigned into survival group and death group according to their clinical outcomes. Binary multivariate unconditional Logistic regression was used to analyze the risk factors of mortality in ELBWI.Results:A total of 637 ELBWI cases were included. 311 cases were in the male group with a survival rate of 57.9% (180/311) and 326 cases were in the female group with a survival rate of 57.4% (187/326). The incidences of neonatal respiratory distress syndrome (RDS), bronchopulmonary dysplasia (BPD), pulmonary hemorrhage and severe intraventricular hemorrhage (IVH) in the male group were significantly higher than the female group ( P<0.05). Significant increases of survival rate existed for both groups year by year ( P<0.01).No significant differences existed in survival rate, mortality rate of infants receiving proactive treatment and mortality rate of infants withdrawing treatment between the two groups ( P>0.05). Multivariate unconditional Logistic regression analysis showed that withdrawing treatment ( P<0.01) and pulmonary hemorrhage ( P<0.05) were associated with increased risks of mortality. Conclusions:Male ELBWI have higher risks of RDS, BPD and severe IVH than female ELBWI. Withdrawing treatment and pulmonary hemorrhage are common risk factors of mortality for both male and female ELBWI.

Objective:To investigate the effect of long non-coding RNA (lncRNA) tumor susceptibility candidate gene 9 (CASC9) on the proliferation and apoptosis of pancreatic cancer cell BxPC-3, and to identify the targeting relationship between miR-195-5p and CASC9.Methods:40 pairs of pancreatic cancer tissues and adjacent normal pancreas tissues resected by surgery and diagnosed by histopathology in Xiangyang Hospital of Integrated Traditional and Western Medicine from April 2017 to May 2018 were collected. Four pancreatic cancer cells (AsPC-1, HPAC, BxPC-3, PANC1) and normal pancreatic ductal epithelial cells HPDE6-C7 were used in experiments. The expression level of CASC9 in pancreatic cancer tissues and cell lines were detected by real-time quantitative PCR. The BxPC-3 cells were divided into si-CASC9 group (transfected with siRNA against CASC9), si-control group (transfected with siRNA that did not match CASC9), CACS9 group (transfected with CASC9 overexpressed plasmid), and CASC9/miR-195-5p group (co-transfected with CASC9 overexpressed plasmid and miR-195-5p mimics). Cell proliferation activity was detected by MTT assay. Western blot was used to detect the protein expression of Bax and Bcl-2. The targeting relationship between CASC9 and miR-195-5p was identified by bioinformatics analysis and luciferase assay.Results:The expression level of CASC9 in pancreatic cancer tissues was significantly higher than that in adjacent normal tissues (4.7±1.25 vs 2.15±0.82, P=0.04), and the expression levels of CASC9 in pancreatic cancer cell lines AsPC-1, HPAC, BxPC-3, and PANC1 cells were 1.43±0.12, 1.86±0.13, 2.03±0.14, and 1.73±0.15, respectively, which were significantly higher than that in HPDE6-C7 cells (1.00±0.10, P<0.001). The expression in BxPC-3 cells was the highest. The proliferation activity of cells in si-CASC9 group decreased significantly compared with that in si-control group (on day 3 0.57±0.05 vs 0.72±0.04, P=0.01; and on day 4 0.75±0.07 vs 0.95±0.07, P=0.02). Bax expression was up-regulated (1.39±0.13 vs 1.07±0.11, P=0.03), while Bcl-2 expression was significantly down-regulated (1.44±0.11 vs 1.71±0.12, P=0.04). The cell proliferation activity of CASC9/miR-195-5p group was significantly decreased compared with that of CASC9 group ( P<0.005). The expression level of Bax was significantly higher than that of CASC9 group (0.68±0.04 vs 0.56±0.03, P=0.01), and the expression level of Bcl-2 was significantly lower than that of CASC9 group (1.05±0.03 vs 1.47±0.04, P<0.001). Conclusions:miR-195-5p can reverse the effect of CASC9 on promoting proliferation and inhibiting apoptosis of pancreatic cancer cells by targeting lncRNA CASC9.

OBJECTIVE: To investigate the mechanism of calcium ion(Ca~(2+))/calcineurin(CaN) signaling pathway in bisphenol A(BPA)-induced interleukin-6(IL-6) secretion in macrophages. METHODS: Raw 264.7 cells in logarithmic growth phase were divided into control group, activator group, BPA low-, medium-and high-dose groups and inhibitor group. The cells in control group were treated with 0.10% dimethyl sulfoxide. The activator group was treated with lipopolysaccharide at mass concentration of 2 mg/L. The 3 BPA groups were treated with BPA at final concentrations of 1, 10, or 100 μmol/L. Two sets of verapamil or tacrolimus(FK506) groups were given verapamil at the final concentration of 10 or 30 μmol/L; or final concentration of FK506 at 250 or 500 nmol/L. Then the cells were treated with final concentration of 100 μmol/L BPA. The cells were collected at 4, 12, and 24 hours. The mRNA expression of IL-6 and CaN at 4 and 12 hours were detected by real-time fluorescence quantitative polymerase chain reaction. The relative expression of IL-6 and CaN protein was detected at 24 hours by enzyme-linked immunosorbent assay, and the intracellular Ca~(2+) level was detected at 4 hours using a single-tube multi-function detector. RESULTS: At 12 hours, the mRNA expression of IL-6 in the 100 μmol/L BPA group was higher than that in control group, activator group and the 1 and 10 μmol/L BPA groups(P<0.05), and higher than that in the 10 and 30 μmol/L verapamil groups, and in the 250 and 500 nmol/L FK506 groups(P<0.05). The mRNA expression of CaN in the 100 μmol/L BPA group was higher than that in control group, activator group and 1 and 10 μmol/L BPA groups(P<0.05), and higher than in 10 and 30 μmol/L verapamil groups(P<0.05). The relative expression of IL-6 protein in the 100 μmol/L BPA group was higher than that in control group, activator group and 1 and 10 μmol/L BPA groups(P<0.05). The relative expression of CaN protein in 100 μmol/L BPA group and 10 and 30 μmol/L verapamil groups were higher than that in control group and activator group(P<0.05). The relative expression of CaN protein in the 10 and 30 μmol/L verapamil groups were lower than that in the 100 μmol/L BPA group(P<0.05). The intracellular Ca~(2+) level in the 100 μmol/L BPA group was higher than that in control group and activator group(P<0.05). The intracellular Ca~(2+) level in the 10 μmol/L verapamil group was lower than that in the 100 μmol/L BPA group(P<0.05). CONCLUSION: BPA might promote the secretion of IL-6 through Ca~(2+)/CaN signaling pathway in macrophages.

Objective: To investigate the regulation of emodin on microRNA expressions in mouse model with sepsis by GeneChip microRNA array. Methods: Forty two c57 mice were randomly (random number) divided into 3 groups: sham operation group (sham group, n=14),sepsis group(n=14) and emodin group (n=14). The sepsis model of the mouse was subjected to cecal ligation and puncture (CLP). Mice in emodin group received intraperitoneal injection of emodin (40 mg/kg) half an hour before operation and every 12 hours after CLP. All mice were sacrificed 48 h after surgery and part of the ileum were removed for intestine tissue stained with hematoxylin eosin. The levels of tumor necrosis factor-α(TNF-α), interleukin-6 (IL-6) and intestinal fatty acid binding protein (I-FABP) in peripheral blood were measured by enzyme-linked immunosorbent assay (ELISA). Total RNA of ileum tissues was extracted from the sepsis group and emodin group, and then subjected to miRNA microarray. The results of microarray were further verified by quantitative real-time PCR (qRT-PCR). Target genes of differentially expressed miRNAs were predicted and subjected to gene ontology (GO) and KEGG pathway enrichment analysis. Data of multi-groups were analyzed by one way variance (ANOVA) and inter-group comparisons were made by SNK-q tests. Mann-Whitney U test was used when homogeneity of variance were not met. The value of P<0.05 was considered statistically significant. Results: Compared to the sepsis group, the levels of serum TNF-α, IL-6 and I-FABP in the emodin group were decreased significantly. MiRNA microarray showed that 23 miRNAs were differentially expressed in the emodin group as compared to the sepsis group, among which 17 miRNAs were up-regulated and 6 miRNAs were down-regulated. qRT-PCR results were consistent with miRNA array data. Target genes of 10 selected miRNAs were predicted and subjected to bioinformatic analysis. A total of 3 410 target genes were significantly enriched in 2 072 GO biological process terms, 246 GO cellular component items and 277 GO molecular function terms. Moreover, KEGG pathway analysis showed that these differential miRNAs were involved in the regulation of PI3K-Akt signaling pathway, MAPK signaling pathway, and TNF signaling pathway. Conclusions Emodin can regulate the expression of multiple microRNAs, and play an important role in protecting the intestinal mucosal barrier via a variety of targets and pathways.

Objective::To explore the effects of the biomimetic network membrane prepared by chitosan/gelatin/pectin on the proliferation and mineralization of mesenchymal stem cells (MSCs) , and to evaluate its feasibility of constructing tissue engineering bone.Methods:Chitosan, gelatin and pectin were made into a new biomimetic network membrane in a certain ratio by biomimetics.The experiment was divided into control group (MSCs+conventional medium) , material group (MSCs+network membrane+conventional medium) and material+OS group (MSCs+network membrane+OS medium) .The cell morphology was observed by inverted phase contrast microscope;the growth and secretion of extracellular matrix of the MSCs were observed under scanning electron microscope (SEM) .The proliferation of cells was determined by MTT assay (The MSCs were divided into negative control group and material group, and they were cultivated with blank medium and medium including materials) .The expression of calcium in MSCs was detected by Alizarin Red staining.Real-time polymerase chain reaction (RT-PCR) was used to determine the expression levels of osteocalcin (OC) mRNA and osteopontin (OPN) mRNA in the MSCs.Results:The network membrane was semitransparent thin film.The MSCs were short shuttle and clustered under inverted phase contrast microscope.After cultured for 7d, the MSCs were shuttle;after cultured for 14d, the number of MSCs was increased, with pseudo feet on the membrane;after cultured for21d, the MSCs clustered with a lot of neo-formed extracellular matrix.The MTT results showed that there was no significant difference in the proliferation level of MSCs between material group and negative control group (P＞0.05) .The Alizarin Red staining results showed that the MSCs in the network membrane were dyed orange red.The RT-PCR results showed that the expression levels of OC mRNA in the MSCs in material group and material+OS group were lower on the 7th and 14th days, but on the 21th day, the expression levels were significantly increased and reached the peak;the expression level of OC mRNA in the MSCs in material group was significantly increased on the 7th day, and the expression level reached the peak on the 14th day, then fell slightly on the 21th day;compared with control group, the expression levels of OC mRNA and OPN mRNA in the cells in material group and material+OS group at different time points were significantly increased (P＜0.01) , but there were no significant differences between material group and material+OS group (P＞0.05) .Conclusion:Chitosan/gelatin/pectin biomimetic network membrane has good biocompatibility, and MSCs can grow and proliferate well on the membrane.The membrane can induce the MSCs to express mineralization-related genes and proteins without inducers.

Objective: To investigate the clinical and histological features, diagnosis and differential diagnosis of myofibroma/myofibromatosis. Methods: The clinical data and pathology features of nine cases of myofibroma/myofibromatosis were collected from August 2011 to November 2016 in Affiliated Drum Tower Hospital, Nanjing University Medical School and Children′s Hospital of Nanjing Medical University. Immunohistochemistry(IHC), PDGFRB molecular analysis and ETV6-NTRK3 gene fusion were performed and relevant literature reviewed. Results: There were 7 males and 2 females, with age ranging from 3 days to 18 years (mean 5 years). The tumors were located in head and neck (eight cases) and trunk (one case). Clinically, the tumors presented as freely movable nodules. Microscopically, they appeared biphasic with alternating light- and dark-staining areas. The light-staining area consisted mainly of plump myoid spindle cells with eosinophilic cytoplasm arranged in nodules, short fascicles, or whorls.The dark-staining area was composed of round or polygonal cells with slightly hyperchromatic nuclei or small spindle cells arranged around a distinct hemangiopericytoma-like vascular pattern. IHC showed the tumor cells in the light-staining area were strongly positive for vimentin and SMA, while cells in dark-staining area were strongly positive for vimentin, and weakly for SMA. Tumor cells were negative for desmin, S-100 protein, h-Caldesmon, CD34 and STAT6. Analysis of PDGFRB mutations was performed in seven cases. Two cases showed 12 exon point mutation c. 1681 c>T(p.R561C), one case showed 14 exon point mutation c. 1998C>G (p.N666K). ETV6-NTRK3 gene fusion was not detected by fluorescence in situ hybridization in four patients under three years old. All cases were followed for 6 to 68 months, with two recurrences. Conclusions Myofibroma/myofibromatosis is an uncommon benign myofibroblastic tumor of infancy and childhood. The tumor can appear biphasic, and may show PDGFRB point mutation which is of potential diagnostic value.

Objective To investigate the effect of Danggui buxue decoction on ventricular remodeling and miR-34a expression after myocardial infarction in mice. Methods Mice myocardial infarction model were estab-lished by the left anterior descending coronary artery embolization. Then the mice were divided into Danggui buxue Decoction group and model group. Mice in Danggui buxue decoction group were given Danggui buxue decoction , and mice in the model group were given the equal volume of saline for 8 weeks. Cardiomyocyte apoptosis was detect-ed by TUNEL staining. Myocardial fibrosis was detected by Masson staining. The expression of miR-34a was detect-ed by quantitative PCR and the expression of Sirt1 was detected by Western blot assay. Results Compared with the model group ,the EF value of the mice was significantly improved (P < 0.05),the myocardial cell index decreased(P<0.05),the content of collagen fibers decreased(P<0.05),the expression of miR-34a decreased, but the expression of Sirt1 increased(P<0.05)in Danggui buxue Decoction group. Conclusion Danggui buxue decoction can effectively inhibit ventricular remodeling of mice following myocardial infarction ,which is associated with the suppression of miR-34a and the increase of Sirt1.

Objective To study the toxicokinetics of Cymermethrin and its metabolites in dog bile and provide evidence for forensic cases of identification of Cymermethrin poisoning. Methods 1/4LD50 doses of Cymermethrin were given to 6 male dogs by oral perfusion after the gallbladder fistula surgery on them,and their bile were collected at different time, in which Cymermethrin and its metabolites were extracted by Liquid-liquid extraction with dichloromethane and detected by HPLC-MS-MS. The qualitative analysis was based on retention time and MRM ions. The quantitative analysis was based on an internal standard method and calibration curve. Toxicokinetics equations of Cymermethrin and its metabolites in the bile were established from the c-t curves which were fitted by the WinNonlin toxicokinetics software meanwhie toxicokinetics parameters were obtained. Results The toxicokinetics of Cymermethrin met first-order dynamic equation. The Tmax of Cymermethrin(CYM), 3-phenoxybenzoic acid (3-PBA), 3-(2,2-Dichloroethenyl)-2,2-dimethyl-cyclopropanecarboxylate (DCVA) respectively were 1.52±0.30,1.29±0.04,0.93±0.41 h ; The Cmax of CYM, 3-PBA, DCVA respectively were 0.38±0.03,7.9±1.32,30.9±16.24 μg/mL ; The T1/2 of CYM, 3-PBA, DCVA respectively were3.93±0.71,1.36±0.11,4.49±2.81 h; Conclusion The toxicokinetics of Cymermethrin in dog bile met first-order dynamic equation ; The toxicokinetics model and parameters of Cymermethrin can provide evidence for forensic identification of Cymermethrin poisoning cases.