Melatonin alleviates alcoholic liver disease via EGFR-BRG1-TERT axis regulation

Chronic alcohol consumption causes liver steatosis, cell death, and inflammation. Melatonin (MLT) is reported to alleviate alcoholic liver disease (ALD)-induced injury. However, its direct regulating targets in hepatocytes are not fully understood. In the current study, a cell-based screening model and a chronic ethanol-fed mice ALD model were used to test the protective mechanisms of MLT. MLT ameliorated ethanol-induced hepatocyte injury in both cell and animal models (optimal doses of 10 μmol/L and 5 mg/kg, respectively), including lowered liver steatosis, cell death, and inflammation. RNA-seq analysis and loss-of-function studies in AML-12 cells revealed that telomerase reverse transcriptase (TERT) was a key downstream effector of MLT. Biophysical assay found that epidermal growth factor receptor (EGFR) on the hepatocyte surface was a direct binding and regulating target of MLT. Liver specific knock-down of Tert or Egfr in the ALD mice model impaired MLT-mediated liver protection, partly through the regulation of nuclear brahma-related gene-1 (BRG1). Long-term administration (90 days) of MLT in healthy mice did not cause evident adverse effect. In conclusion, MLT is an efficacious and safe agent for ALD alleviation. Its direct regulating target in hepatocytes is EGFR and downstream BRG1-TERT axis. MLT might be used as a complimentary agent for alcoholics.

Clinical and dermoscopic manifestations of nail damage in 12 cases of hand, foot and mouth disease / 中华皮肤科杂志

Objective:To investigate clinical and dermoscopic manifestations of nail damage secondary to hand, foot and mouth disease.Methods:Clinical data were collected from 12 patients with nail damage secondary to hand, foot and mouth disease in Department of Dermatology, Shandong Provincial Hospital from June to November 2015, and characteristics of nail damage were analyzed.Results:The clinical manifestations of nail damage following hand, foot and mouth disease included dryness of periungual skin (12 cases) , nail plate cavities (11 cases) , nail fractures (11cases) and Beau′s lines (4 cases) . Dermoscopy showed nail discoloration (12 cases) , nail delamination (12 cases) , horizontal streaks (10 cases) , longitudinal streaks (8 cases) , brown background of the nail matrix (11 cases) , telangiectasia (7 cases) , periungual desquamation (12 cases) and red background (10 cases) .Conclusion:The clinical manifestations of nail damage following hand, foot and mouth disease are mainly dryness of periungual skin, nail plate cavities and fractures, and its dermoscopic manifestations include nail discoloration, delamination, horizontal and longitudinal streaks, brown background of the nail matrix, telangiectasia, periungual desquamation and red background.

Sedative efficacy of remimazolam in outpatients undergoing curettage / 中华麻醉学杂志

Objective:To evaluate the sedative efficacy of remimazolam in outpatients undergoing curettage.Methods:Eighty outpatients, of American Society of Anesthesiologists physical status Ⅰor Ⅱ, aged ≥18 yr, undergoing curettage, were divided into 2 groups according to the random number table method: remimazolam group (group R, n=41) and propofol group (group P, n=39). Anesthesia was induced with alfentanil 10 μg/kg injected intravenously in two groups, and in addition remimazolam 7 mg was intravenously injected in group R, and propofol 1.5 mg/kg was intravenously injected in group P. If the Modified Observer′s Assessment of Alertness/Sedation score ≥3 or the patient could not tolerate the surgical procedure, remimazolam 2.5 mg was given for rescue sedation in group R and propofol 0.5 mg/kg was given for rescue sedation in group P, and alfentanil 1 μg/kg was given as rescue analgesic.The onset time of sedatives, time to eye opening, emergence time and time to discharge were recorded.The success of sedation, intraoperative BIS value, requirement for rescue medications, and occurrence of adverse reactions were recorded. Results:The intraoperative BIS values were maintained at 61-72 and 40-64 in group R and group P, respectively.The success rate of sedation was 95% in group R and 100% in group P, and there was no significant difference between the two groups ( P>0.05). Compared with group P, the rate of rescue sedative agents used and rate of rescue alfentanil given were significantly increased, the onset time of sedative agents was significantly prolonged, the incidence of postoperative nausea and vomiting was increased, and the incidence of respiratory depression and injection pain was decreased in group R ( P<0.05 ). Conclusion:Remimazolam (given according to the instructions) is safer when used for outpatient curettage, but it can only maintain a light sedation status, and the sedative efficacy is inferior to propofol.

Progress in localization of pulmonary nodules / 国际外科学杂志

With the development of CT and the popularization of health examination, the detection rate of small pulmonary nodules has been improved. Some small pulmonary nodules could be malignant nodules. Surgical resection is the preferred treatment. Therefore, it is an important task for thoracic surgeons to accurately locate pulmonary nodules during surgery and remove nodules accurately on the premise of maximum protection of lung function. At present, the core of preoperative auxiliary localization of pulmonary nodules is the implantation of markers. The commonly used clinical localization methods include hook wire localization, microcoil localization, methylene blue puncture injection localization and biological glue localization. In this paper, the development status, application scope, advantages and disadvantages of existing localization methods are briefly reviewed, which can provide references for clinical application and follow-up research.

Association of advanced oxidation protein products in seminal plasma with teratospermia and outcome parameters of fertilization / 南方医科大学学报

OBJECTIVE@#To study the association of the level of advanced oxidation protein products (AOPPs) in seminal plasma with teratospermia and the outcome parameters of fertilization (IVF).@*METHODS@#We conducted a cross-sectional study among 272 male patients receiving assisted reproduction treatment in the Center for Reproductive Medicine of our hospital between October, 2018 and March, 2019. The levels of seminal AOPPs and reactive oxygen species (ROS), demographic data, sperm parameters and IVF outcome parameters were analyzed for all the patients. According to the percentage of sperms with normal morphology, the patients were divided before IVF into teratozoospermia group and normal sperm morphology group, and those in teratozoospermia group were further divided into 3 subgroups with mild, moderate and severe teratozoospermia. The patients were also divided on the day oocyte retrieval into 2 groups with fertilizing rates lower (group Ⅰ) and higher (group Ⅱ) than the median rate.@*RESULTS@#We found a significant negative correlation of seminal AOPP level before treatment with the percentage of normal sperm morphology (=0.003) and seminal ROS level (=0.013). The seminal levels of AOPPs (= 0.027) and ROS (=0.036) were significantly elevated in patients with teratospermia, and seminal AOPP level was significantly higher in severe teratospermia group than in mild (=0.019) and moderate (=0.015) teratospermia groups. The seminal levels of AOPPs (=0.003) and ROS (=0.017) on the day of oocyte retrieval were negatively correlated with the fertilization rate in IVF cycles, and the levels of AOPPs (=0.049) and ROS (=0.036) were significantly higher in group Ⅰ than in group Ⅱ.@*CONCLUSIONS@#An elevated level of seminal AOPPs may indicate an increased risk of severe teratospermia and a lower fertilization rate in IVF.

Occupational, educational and residential factors are associated with the outcomes of in vitro fertilization-embryo transfer / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the associations of occupational, educational and residential factors with the outcomes of in vitro fertilization -embryo transfer (IVF-ET).</p><p><b>METHODS</b>A retrospective analysis was conducted among 3998 patients undergoing the first treatment with IVF-ET and 3064 perinatal infants in our center between January, 2010 and June, 2014. The relation of the patients' occupation, educational background, and residence registry with IVF-ET outcomes was analyzed.</p><p><b>RESULTS</b>The risk of abortion in the first trimester was significantly higher in female patients engaged in agriculture (OR=4.319, P=0.047). Occupation and educational background of the patients and their spouse were related with the incidence of ectopic pregnancy following IVF-ET. The total incidence rate of perinatal birth defects was 2.1% in this cohort, and was the highest in Shenzhen (5.5%) but this difference was not statistically significant (P>0.05).</p><p><b>CONCLUSION</b>The patients' occupation and educational background are related with the outcomes of IVF-ET, suggesting the importance of health education and prenatal examination.</p>

Synthesis, biological activity and molecular docking research of N-{(4-oxo-thiochroman-3-yl)phenyl-methyl}acetamide derivatives as α-glucosidase inhibitors / 药学学报

In order to develop potent antidiabetic agents that have inhibitory effect to a-glucosidase, twelve β-acetamido ketone derivatives such as N-{[(substituted-4-oxo-thiochroman-3-yl)phenyl]-methyl}acetamide are designed and synthesized through one-pot Dakin-West reaction. Their chemical structures are confirmed by 1H NMR, 13C NMR, IR and HR-MS. In vitro α-glucosidase inhibition assays of compounds 4a-41 were carried out using glucose oxidase method. The result indicated that most of them possess inhibitory activity in vitro. Compound 4k showed the most potent inhibitory activity with 87.3% inhibition of α-glucosidase at the concentration of 5.39 mmol x L(-1). The structure-activity relationship of these β-acetamido ketone derivatives was discussed preliminarily. Moreover, the molecular docking method was used to study the interaction mode of compound 4k and α-glucosidase. Our results will be helpful for designing of α-glucosidase inhibitors in the future.

Construction of HaCaT cell lines stably expressing the human GJB6 gene by using a Tet-On lentiviral vector and their identification / 中华皮肤科杂志

Objective To construct HaCaT cell lines stably expressing the wild type human GJB6 gene or its mutant by using a Tet-On lentiviral vector, and to lay an experimental foundation for studies on pathogenesis of hidrotic ectodermal dysplasia. Methods The wild-type human GJB6 gene and its mutant (A88V)were amplified by PCR, and then inserted into the Tet-on lentivirus plasmid to construct recombinant lentivirus vectors. The recombinants were identified by gene sequencing and enzymatic digestion. Cultured HaCaT cells were classified into three groups to be transfected with a negative control lentiviral vector (NC group), the lentivirus vector expressing the wild-type human GJB6 gene (WT group), or the lentivirus vector expressing the mutant human GJB6 gene (MU group). Puromycin was used to select HaCaT cell clones stably expressing the GJB6 gene which encodes the connexin 30 (Cx30)protein. The selected HaCaT cell clones were cultured with or without tetracycline for 48 hours, thereafter, real-time PCR(RT-PCR) was performed to detect GJB6 gene mRNA expression, Western-blot analysis to measure expressions of Cx30 and FLAG-tag proteins, and cell counting kit 8 (CCK8)assay to evaluate cellular proliferative activity. Results Enzymatic digestion and gene sequencing showed that recombinant lentivirus plasmids were successfully constructed. RT-PCR showed evidently increased mRNA expression of the GJB6 gene in stably transfected HaCaT cells. Moreover, the expression abundance of the GJB6 gene was 112.369 times higher in the WT group induced by tetracycline than in that without tetracycline treatment (P 0.05). Conclusion HaCaT cell lines which stably express the wild-type GJB6 gene or its mutant(A88V)are successfully constructed.

Mutation analysis of GJB2 gene in a family with Vohwinkel syndrome / 中华皮肤科杂志

ObjectiveTo detect the mutation of GJB2 gene in a Chinese family with Vohwinkel syndrome.MethodsClinical data were collected from 5 patients with Vohwinkel syndrome in a family,and blood samples were obtained from the 5 patients and 4 unaffected individuals in the family as well as from 100 normal human controls.Genomic DNA was extracted and subjected to PCR for the amplification of the entire encoding and flanking sequences of GJB2 gene(1015 bp) followed by bidirectional sequencing with the ABI PRISM 3730 automatic DNA sequencer.Finally,sequence alignment was carried out by using the software Sequencher 4.10.1 Demo.ResultsA heterozygous missense mutation 196G→C in the GJB2 gene,which resulted in the substitution of aspartic acid by histidine at codon 66 (D66H) in the first extracellular domain of the protein,was observed in all the patients of this family,but in none of the 4 unaffected individuals in this family or the 100 normal human controls.ConclusionThe D66H missense mutation in the GJB2 gene may contribute to the occurrence of Vohwinkel syndrome in Chinese Han population.

Effects of 5-aza-dc on the expression of insulin-like growth factor binding protein 7 (IGFBP7) in and proliferation of melanoma cells / 中华皮肤科杂志

Objective To study the effects of 5-aza-dc on the expression of IGFBP7 in and proliferation of melanoma cell lines A375 and M14.Methods Reverse transcription-PCR and immunocytochemistry were performed to detect the mRNA and protein expression of IGFBP7 in A375 cells and M14 cells after treatment with 5-aza-dc of 10μmol/L for 48 hours,and MTT assay to measure the proliferation of both cell lines treated with 4 different concentrations (2.5,5,10,20μmol/L) of 5-aza-dc for various durations.Results The treatment with 5-aza-dc restored IGFBP7 expression at both mRNA and protein levels.The four concentrations of 5-aza-dc inhibited the proliferation of A375 and M14 cells in a dose-dependent (F=561.12,271.43,respectively,both P<0.01) and time-dependent (F=141.35,549.33,respectively,both P<0.01) manner.Conclusions DNA methylation may be involved in the modulation of aberrant IGFBP7 gene expression in melanoma,and 5-aza-dc could inhibit the proliferation of A375 and M14 cells.

In vitro anti-proliferation effect of a histone deacetylase inhibitor,chidamide,on a malignant melanoma cell line,A375 / 中华皮肤科杂志

Objective To investigate the in vitro anti-proliferation effect of a histone deacetylase inhibitor,chidamide,on a cutaneous malignant melanoma cell line,A375.Methods Cultured A375 cells were treated with different concentrations of chidamide(5,10,50,100,500 μmol/L)and aichostatin A (TSA)(0.1,0.25,0.5,1.0 μmol/L),respectively,for various durations(24,48,72,96,120 hours).Subsequently,cell proliferation,apoptosis and cell cycle were detected by MTT assay,annexin Vfluorescein isothiocyanate and propidium iodide double staining,and DNA ploid analysis,respectively.Results The proliferation of A375 cells was inhibited in a dose-dependent manner by chidamide of 5-500μmol/L and TSA of 0.1-1 μmol/L,and in a time-dependent manner from 0 to 120 hours after the beginning of trealment with ehidamide of 5-500μmol/L and TSA of 0.25-1μmol/L.The 48-hour 50% growth inhibition concentration(IC50)of ehidamide and TSA on A375 cells was about 250 μmol/L and 0.7μmol/L,respectively.After 48-hour treatment,the apoptosis mte was 80.27%±3.06%,79.53%±5.70%,83.13%±6.90%in A375 cells treated with chidamide of 62.5,125,250 μmol/L,respectively,16.27%±2.46%,28.83%±2.55%,83.40%±8.65%in those treated with TSA of 0.175,0.35,0.7 μmol/L,respectively,10.43%±0.96%in ontreated cells;a statistical increase was noticed in chidamide-treated cells and TSA-treated cells vs.untreated cells(all P＜0.001).A positive correlation was observed between the apoptosis rate and concentrations of TSA(r=0.955,P=0.000).Cell cycle analysis indicated that treatment with chidamide induced cell cycle arrest in G0/G1 phase,with the cell proportion in G0/G1 phase being 76.30%±6.06%,82.79%±0.74%,88.91%±5.29%in A375 cells treated with chidamide of 62.5,125,250μmol/L,respectively,versus 38.73%±3.36%in untreated cells.While after 48-hour treatment with TSA of 0.35 and 0.7 μmol/L,the proportion of cells in G2/M phases was 25.15%±2.71%and 58.71%±3.45%,respectively,compared to 15.73%±0.23%in untreated cells(P＜0.01).Conclusion Chidamide and TSA could induce cell cycle arrest and apoptosis,as well as inhibit the growth of A375 ceils in vitro.

Gene diagnosis in a family with hidrotic ectodermal dysplasia / 中华皮肤科杂志

Objective To confirm the diagnosis and to localize the pathogenic gene of ectodermal dysplasia in a family SUffering from only hair and nail abnormalities.MethodsBlood samples were collected from 7 affected patients and 15 unafiected individuals in the family.Genomic DNA was extracted from blood samples by routine phenol-chloroform methods.The whole coding regions of candidate genes K16,K17,K6a,K6b and GJB6 were amplified by PCR followed by direct sequencing.Then,the gene mutation was further confirmed at mRNA level by RT-PCR.ResultsA heterozygous missense mutation 3 1G→A in the GJB6 gene.which leads to the substitution of glycine by arginine at codon 11(G11R)on the N-terminal of the protein,was detected in all the patients.but in none of the 15 normal individuals in this family.The mutation was also confirmed in the CDNA originating from the proband's skin biopsy.Conelusionn A missense mutation G31A.which has been shown previously to cause hidrotic ectodermal dysplasia(HED),is localized in the GJB6 gene of patients in this family.

Silencing BAO-1 gene by eukaryotic expression vector containing short hairpin RNA in mouse melanoma B16F10 cells / 中华皮肤科杂志

Objective To construct the eukaryotic expression plasmids of short hairpin RNA (shRNA) specific for mouse Bcl-2-assoeiated athanogene 1 (BAG-1) and to observe their inhibitory effects on the expression of BAG-1 gene in mouse melanoma B16FI0 ceils. Methods Plasmids named pRNAT-U6.1/Neo-BAG-1, were designed and constructed to target the mouse BAG-1 mRNA coding region. LipofectaminTM 2000 was used to transfect plasmids into BI6F10 cells. Negative plasmid-transfected and tmtransfected B16F10 cells served as negative and blank controls respectively. Forty-eight hours following transfection, G418 was used to select the resistant cells. The mRNA and protein expression of BAG-1 gene was measured by reverse transcription-PCR and Western blot respectively about 1 month after the transfection. Results The eukaryotic expression plasmids, pRNAT-U6.1/Neo-BAG-1, were constructed, and verified by restriction enzyme digestion and DNA sequencing. The transfection rate in B16F10 cells was 20% -30%. Compared with the blank control, the mRNA and protein expression of BAG-1 in BI6FI0 cells was significantly inhibited by BAG-1 shRNA (both P<0.05), and the inhibition rates were (77±4)% and (62 ±2)%, respectively. Conclusions These results indicate that the eukaryotic expression vectors containing shRNA against BAG-1 gene, pRNAT-U6.1/Neo-BAG-1, are successfully constructed, and can significantly inhibit the expression of BAG-1 gene in mouse melanoma B16F10 cells.