Post-translational modification by ubiquitin and ubiquitin-like modifiers(Ubls) is one of the most important mechanisms regulating a wide range of cellular processes in eukaryotes.Previous research showed that,through covalently modification by ubiquitin or ubls,the substrate proteins can be regulated in many different ways like stability,subcellular localization,enzymatic activity,protein-protein interaction and so on.Therefore,we believe,that ubiquitin and ubls play very important roles in cellular and biological processes by modifying plenty of proteins.To better understand the ubiquitin and ubls system,proteomic approaches have been developed to purify and identify more protein substrates.Large-scale idendification of ubiquitin/ubls-modification sites by mass spectrometry is particularly important for understanding the molecular mechanism and function of ubiquitin/ubls modification.Upto the present,more and more scientists are getting interested and participating in proteomics research of ubiquitin/Ubl modifications.This review summarizes the rencent results in this field.

The event-related potential (ERP) is considered as one of the most effective methods to study and analyze objectively human mental activity based on nerve electrophysiology. At present, ERP is not only used in the study of lie detection, but also in the clinical medicine for the cognitive assessment on patients with cerebrovascular disease, dementia or traumatic brain injury and auxiliary diagnosis of mental illness. With the further development of ERP detection technology, it would have a wider application prospect in the field of forensic medicine.
Evoked Potentials/physiology* ; Forensic Medicine/trends* ; Humans

Objective To investigate the efficacy of zoledronic acid in the treatment of senile unstable femoral intertrochanteric fractures with anatomical locking plate. Methods 67 patients were randomly divided into two groups. Five days after the operation, group A received one intravenous injection of 5 mg zoledronic acid, while patients in group B did not receive the injection. The two groups were compared in terms of hospitalization time, complications, limb weight-bearing time, fracture healing time, hip function score after operation, preoperative and postoperative serum calcium and serum ALP, bone mineral density of proximal femur before operation and 1 year after operation. Results There were no statistically significant differences between the two groups in age, type of fracture, hospital stay, partial weight-bearing time, fracture healing time, hip function at 1 month and 1 year after operation, preoperative bone mineral density and blood calcium. But the differences were statistically different in hip function at 3 months after operation , averaged bone mineral density of proximal femur and serum ALP 1 year after operation. Moreover, 5 patients in group A developed muscle pain or fever after intravenous injection of zoledronic acid. Conclusion The locking plate combined with zoledronic acid injection in treatment of elderly patients with unstable femoral intertrochanteric fracture could inhibit bone loss, increase bone mineral density, and accelerate limb function recovery after operation. On the other hand, Zoledronic acid has a high incidence of adverse reaction.

On 19 December 1997, SilkAir Flight MI 185, a Boeing B737-300 airliner crashed into the Musi River near Palembang, Southern Sumatra, enroute from Jakarta, Indonesia to Singapore. All 104 passengers and crew onboard were killed. Of the human remains recovered, 6 positive identifications were made, including that of one Singaporean. Two of the identifications were by dental records, 2 by fingerprints, 1 by age estimation and 1 by personal effects. This paper describes the crash victim identification of Flight MI 185. The authors were part of an Indonesia- Singapore forensic team deployed for 3 weeks in Palembang to assist the Indonesian authorities in human remains identification.
Accidents, Aviation ; Child, Preschool ; Dental Records ; Dermatoglyphics ; Female ; Forensic Dentistry ; Humans ; Indonesia ; Male

In this study, we explored the mechanism of Huganning tablet (HGNP) in the treatment of nonalcoholic fatty liver disease (NAFLD) based on network pharmacology and computer-aided drug design. Firstly, the potential ingredients and targets of HGNP were identified from TCMSP database, Swiss Target Prediction database, Chinese pharmacopoeia (2015) and literatures, and then the targets of HGNP intersected with NAFLD disease targets that obtained in GeneCards database to acquired potential targets. The bioconductor bioinformatics package of R software was used for gene ontology (GO) enrichment and Kyoto encyclopedia of genes and genomes (KEGG) enrichment analysis. The network of “potential ingredient-key target-pathway” was formed in Cytoscape software to study the interactions between potential ingredients of HGNP, key targets, pathways and NAFLD. Based on the results of network pharmacology, the molecular docking analysis of the key targets and potential active ingredients in HGNP tablets with top degree in the network was conducted using Discovery Studio 2020 software, followed by molecular dynamics simulations, binding free energy calculation, drug-likeness properties analysis and ADMET (absorption, distribution, metabolism, excretion and toxicity) properties prediction. In vitro, HepG2 cells were used to establish steatosis model, and the effects of five key compounds on hepatocyte steatosis were analyzed by oil red O staining and triglyceride (TG) content determination. The results showed that 141 ingredients and 151 potential targets were obtained. A total of 2 526 items and 151 pathways were identified by GO and KEGG enrichment analysis. The molecular docking suggested that five components, isorhamnetin, salvianolic acid B, emodin, resveratrol and rhein, exhibited strong binding ability with key targets [retinoic acid receptor RXR-alpha (RXRA), tumor necrosis factor (TNF), glycogen synthase kinase-3 beta (GSK3B), serine/threonine-protein kinase 1 (AKT1)]. It was further verified that isorhamnetin and salvianolic acid B bind to key targets with good structural stability and binding affinity based on molecular dynamics simulations and binding free energy calculations. The drug-likeness properties, pharmacokinetic properties and toxicity of five key compounds were more comprehensively analyzed through drug-likeness properties analysis and ADMET properties prediction. In vitro, all five compounds, isorhamnetin, salvianolic acid B, emodin, resveratrol, and rhein, improved hepatocyte steatosis of HepG2 cells, confirming the reliability of the present study. In conclusion, based on network pharmacology, computer-aided drug design and in vitro validation, this study investigated the mechanism of HGNP for the treatment of NAFLD at multiple levels and provided a basis for its clinical application.

OBJECTIVETo investigate the mechanism of tazarotene against active psoriasis vulgaris.

METHODSA randomized, controlled trial was conducted in 43 patients with active psoriasis vulgaris, who were divided into tazarotene and control groups. Promyelocytic leukemia (PML) mRNA in active psoriatic lesions before and 14 days after tazarotene treatment was detected by in situ hybridization.

RESULTSPML mRNA expression was detected not only in the basal layer (86.96%), but also in the suprabasal layers of the epidermis in the manner of focal expression (78.26%). After tazarotene treatment, virtually no PML mRNA expression could be detected in the psoriatic lesions (8.69% in the basal layer and 4.35% in the suprabasal layers). PML mRNA expression in the control group underwent no obvious changes during the observation.

CONCLUSIONSTazarotene may inhibit abnormal proliferation of keratinocytes through down-regulating PML gene expression in active psoriatic epidermis.

Adolescent ; Adult ; Double-Blind Method ; Down-Regulation ; drug effects ; genetics ; Epidermis ; drug effects ; metabolism ; pathology ; Female ; Gene Expression ; drug effects ; Humans ; In Situ Hybridization ; Keratolytic Agents ; administration & dosage ; therapeutic use ; Male ; Middle Aged ; Neoplasm Proteins ; genetics ; Nicotinic Acids ; administration & dosage ; therapeutic use ; Nuclear Proteins ; genetics ; Promyelocytic Leukemia Protein ; Psoriasis ; drug therapy ; genetics ; RNA, Messenger ; biosynthesis ; genetics ; Transcription Factors ; genetics ; Tumor Suppressor Proteins ; genetics

OBJECTIVETo observe the gene silencing and disruption of WNT pathway mediated by the specific shRNA targeted against beta-catenin and its effect on cell proliferation of the human colon cancer cell line Colo205.

METHODSThe shRNA plasmid vectors against beta-catenin were constructed and transfected into Colo205 cells with Lipofectamine 2000. The expression of beta-catenin was detected by RT-PCR and Western blot. Immunofluorescence staining was also performed to detect the beta-catenin protein expression in cells. The cell proliferation inhibition was determined by MTT assay and soft agar colony formation assay.

RESULTSThe shRNA vectors targeted against beta-catenin were successfully constructed and efficiently suppressed the expression of beta-catenin mRNA and protein(P<0.05). The expression inhibition rates were 47.89% and 45.26% at the mRNA and protein level respectively. Immunofluorescence microscopy also demonstrated the inhibition of beta-catenin protein induced by these specific shRNAs. The MTT assay indicated that the specific shRNA resulted in significant inhibition of cell growth on the culture plates in time-dependent manner. At 72 h post-transfection, the cell viability of CAT group was 48.5%, which was significantly different as compared with that of blank control group's 91.3%(P<0.05). In the soft agar, there were 9, 46, 43 cell colonies in the CAT, blank, and negative control groups respectively, which were significantly different(P<0.05).

CONCLUSIONSThe specific shRNAs targeted against beta-catenin has a gene silencing effect and blocks the WNT signaling pathway, which can inhibit the growth of Colo205 cells.

Cell Line, Tumor ; Cell Proliferation ; Colonic Neoplasms ; genetics ; metabolism ; Humans ; In Vitro Techniques ; RNA Interference ; Signal Transduction ; Transfection ; Wnt Proteins ; metabolism ; beta Catenin ; genetics

OBJECTIVETo identify the localization of hair follicles stem cell (HFSC) in different stages of hair and explore the differentiating capacity of HFSC into epidermis in vitro.

METHODSHFSC were detected by K19 immunostaining in normal human skin. Then, the isolated HFSC through enzyme digestion were seeded on dermal equivalent (DE) and cultured between the air-liquid interfaces for 14 days. Skin-equivalents was harvested and used for evaluation.

RESULTSHFSC mainly located in outer root sheet in hair follicle and human anagen hair follicles containing two distinct reservoirs for K19-positive cells located in the bulge and bulb of the follicle. These two reservoirs fused in line of outer root sheets during the catagen-telogen transition phase and individualized again in the newly forming anagen hair follicle. Based on DE, growing HFSC built a multilayered and confined epidermis.

CONCLUSIONHFSC located in outer root sheets can promote hair cycle and differentiate into epidermis in vitro.

Cell Differentiation ; physiology ; Cells, Cultured ; Epidermis ; cytology ; Hair Follicle ; cytology ; Humans ; Stem Cells ; cytology

OBJECTIVETo study the effects of diallyl disulfide (DADS) on apoptosis of human leukemia K562 cells and possible mechanisms.

METHODSThe morphologic changes of leukemia K562 cells after DADS treatment were observed by Hoechst 33258 staining. Cell apoptosis rates after different concentrations and different durations of DADS treatment were determined by flow cytometry. Fas, FasL and caspase-8 mRNA expression was estimated by reverse transcription-polymerase chain reaction (RT-PCR) 48 hrs after DADS treatment.

RESULTSThe characteristics of apoptosis in K562 cells induced by DADS were observed. After 24 hrs of DADS treatment, the apoptosis rate of K562 cells increased from (11.60 ± 0.83)% at the concentration of 10 mg/L to (37.94 ± 0.87)% at the concentration of 40 mg/L. The apoptosis rate of K562 cells increased after 40 mg/L DADS with the increasing time from (37.94 ± 0.87)% (24 hrs) to (47.02 ± 0.66)% (72 hrs). Expression of Fas and caspase-8 mRNA increased, while FasL mRNA expression decreased significantly 48 hrs after DADS treatment compared with the control group (P<0.05).

CONCLUSIONSDADS can induce apoptosis of human leukemia K562 cells in a time- and concentration-dependent manner, possibly through increasing Fas and caspase-8 expression and decreasing FasL expression.

Allyl Compounds ; pharmacology ; Antineoplastic Agents ; pharmacology ; Apoptosis ; drug effects ; Bisbenzimidazole ; Caspase 8 ; genetics ; Disulfides ; pharmacology ; Fas Ligand Protein ; genetics ; Flow Cytometry ; Humans ; K562 Cells ; RNA, Messenger ; analysis ; fas Receptor ; genetics

Antigen presenting is the initial step of the immune responses. In order to verify that human bronchial epithelial cells (HBECs) can express antigen presentation molecules, which can be modulated by intrapulmonary regulatory peptides, the present study was designed to examine the expressions of human leukocyte antigen DR (HLA-DR), CD80 and CD86 in resting or ozone-stressed HBECs by using immunocytochemistry and flow cytometry analysis. The results showed that HBECs expressed HLA-DR, CD80 and the expressions of HLA-DR and CD80 molecules were down-regulated under ozone stress. While VIP, P3513 and CGRP upregulated the expression of HLA-DR in resting or ozone-stressed HBECs, they had different effects on CD80 expression. VIP did not influence the expression of CD80 under resting state, but increased the expression of CD80 under ozone stress. CGRP decreased CD80 expression in resting HBECs, but increased CD80 expression in ozone-stressed HBECs. P3513 increased CD80 expression in resting HBECs, but decreased CD80 expression in ozone-stressed HBECs. The expression of CD86 was absent in resting or ozone-stressed HBECs. The results obtained demonstrate that HBECs have the capability to act as antigen presenting cells and the expression of HLA-DR and costimulatory molecules can be modulated by intrapulmonary regulatory peptides.
Antigen-Presenting Cells ; metabolism ; B7-1 Antigen ; metabolism ; B7-2 Antigen ; metabolism ; Bronchi ; cytology ; Calcitonin Gene-Related Peptide ; pharmacology ; Epithelial Cells ; metabolism ; HLA-DR Antigens ; metabolism ; Humans ; Ozone ; adverse effects ; Vasoactive Intestinal Peptide ; pharmacology