In recent years, the targeted protein degradation technology has developed quickly, with proteolysis-targeting chimera (PROTAC) as the best-known strategy through exploring the ubiquitin-proteasome system. A number of new targeted protein degradation strategies have been emerging to expand the scope of protein degradation technology, including lysosome-targeting chimeras (LYTACs), autophagy-targeting chimeras (AUTACs), autophagosome-tethering compounds (ATTECs) and chimeras based on chaperone-mediated autophagy (CMA). The emerging methodologies have explored another important protein degradation system in eukaryotes-lysosomal systems, such as the endosome-lysosome pathway and the autophagy-lysosome pathway. This review summaries the mechanisms and features of different strategies for targeted protein degradation, with a special emphasis on the new targeted protein degradation technologies, such as their current status, advantages and limitations.

Objective To recommend the safety guidelines for workplace violence on health care sector according to the incidents of violence status on medical workplace.Methods A pilot study was conducted using a two-round Delphi method to study out the safety guidelines for hospital violence.Results In two subsequent rounds,the group discussed and screened out 50 entries from 51 items in the six modules as safety guidelines for hospital violence.Conclusions Establishment of safety guidelines for hospital violence on health care sector using Delphi method requires further clinical validation.

Adult ; Carotid Body Tumor ; complications ; Female ; Glomus Jugulare Tumor ; complications ; Humans

Objective To investigate the clinical significance of anti-nuclear envelope protein antibody (gp210),anti-soluble acid resistant nucleoprotein (sp100) and anti-mitochondrial antibody M2 subtype (AMA-M2) in sjogren syndrome (SS) and primary biliary cirrhosis (PBC).Methods A total of 241 hospitalized patients diagnosed with connective tissue disease (CTD) were recruited.Anti-gp210,anti-sp100 and AMA-M2 were detected by indirect immunofluorescence.Results (1) Positive rate of AMA-M2,anti-sp100 and antigp210 in 241 cases CTD patient were 10.4％ (25/241),3.3％ (8/241) and 2.9％ (7/241) respectively.(2) There were 16 cases with SS,5 cases with SS-PBC overlap syndrome and 17 cases with PBC in 241 patients with CTD.Distinction among groups of PBC,SS,SS overlapping PBC of positive incidence of AMA-M2 antibody (x2 =6.584,P =0.03) and anti-gp210 (x2 =8.735,P ＜ 0.01) were significantly different,while there was no apparent difference about positive rate of anti-sp100 among the three groups (x2 =3.343,P =0.18).(3) Positive expression of either antibody of anti-gp210 or anti-sp100 in the three groups of SS,SS overlapping PBC,PBC were 3 cases,4 cases,4 cases respectively.The positive rates of any of three autoantibodies in three groups of were 8 cases,5 cases,13 cases respectively.(4) There were significant difference in terms of serum ALB(t =3.858,P＜0.000 1),TSB(t =5.473,P＜0.000 1),ALT(t =2.235,P=0.026),AKP(t =3.141,P =0.002) and γ-GT (t =2.317,P =0.021) in liver damaged patients of all CTD between AMA-M2 positive and negative patients (P ＜ 0.05).However,serum TSB in anti-sp100 positive and negative patients were differed (t =7.892,P ＜ 0.000 1).Serum AKP was different between anti-gp210 positive and negative patients (t =2.451,P =0.015).Conclusion Positive rate of anti-gp210,anti-sp100 and AMA-M2 are the highest in patients with SS overlap of the PBC among CTD patients.Combined detection can improve the sensitivity of diagnosis.Antisp100 and anti-gp210 are valuable for the diagnosis of SS-PBC overlaps syndrome with negative AMA-M2.

The expression of N-myc down-regulated gene 1 (NDRG1) has previously been reported to be involved in the proliferation, differentiation, invasion and metastasis of cancer cells, but its role in cervical cancer is still unclear. This study aimed to investigate the expression of NDRG1gene in human cervical cancer and its effect on aggressive tumor behaviors. The NDRG1 expression in cervical tissues and cells was detected by RT-PCR. Specific expression plasmid pEGFP-N1-NDRG1-GFP was used to enhance the expression of NDRG1 in human cervical cancer cell lines. The mRNA and protein level of NDRG1 was assessed by RT-PCR and Western blotting, respectively. Its effects on cell proliferation, migration, invasion, cell cycle and apoptosis were detected by MTT, transwell migration assay and flow cytometry (FCM), respectively. The results showed that the expression of NDRG1 in cervical cancer tissues and cells was significantly lower than in normal cervical tissues (P<0.001). After transfection with pEGFP-N1-NDRG1-GFP, the mRNA and protein expression of NDRG1 was up-regulated in Siha cells, which suppressed cell proliferation (P<0.001), induced cell cycle arrest (P<0.05), reduced invasion and migration of Siha cells (P<0.05), but caused no cell apoptosis. Moreover, vascular endothelial growth factor (VEGF), a tumor-induced angiogenesis factor, was markedly reduced and E-cadherin, a cell adhesion molecule, was increased in the cells transfected with pEGFP-N1-NDRG1-GFP. It was concluded that up-regulated NDRG1 may play a role in the suppression of malignant cell growth, invasion and metastasis of human cervical cancer.

Objective To investigate the expression of microRNA 27a (miR-27a) and relationship with drug resistance in human ovarian cancer A2780/Taxol cells.Methods A stem-loop-mediated real-time PCR was used to detect miR-27a expression in A2780 and A2780/Taxol cells.The cells were transfected with the mimics or inhibitors of miR-27a or negative control RNA ( NC) by lipofectamine 2000.The expressions of MDR1 gene,P-glycoprotein (P-gp) and homeodomain-interacting protein kinase 2 (HIPK2) protein levels were measured by real-time PCR and western blot respectively.Methyl thiazolyl tetrazolium (MTT) assay was used to analyze drug sensitivity.Apoptosis analysis was measured by fluorescence activated cell sorter ( FACS).Results (1) miR-27a was an average of 2.2-fold higher expression level in A2780/Taxol cells than that in A2780 cells,with a significant difference between the two groups (P <0.05).(2) A2780/Taxol cells transfection with inhibitors of miR-27a showed that the levels of MDR1 mRNA was decreased by 39%,P-gp protein level[(26 ±5)%]decreased than that in the NC group[(43 ±7)%],HIPK2 protein level[(30 ±6)%]increased than that in the NC group[(19 ±4)%],the 50% inhibitionconcentration (0.5 (μmol/L) was less than that in the NC group (6.8 μmol/L),apoptosis rate[(32.5 ± 3.6) %]was higher than that in the NC group[(5.6 ±2.1) %],and there were significant differences between two groups (all P < 0.05 ).( 3 ) Transfection of A2780 cells with mimics of miR-27a led to increase MDR1 mRNA expression by 121% as compared with one transfection with NC (P<0.05).Conclusion The expression of miR-27a is upregulated in A2780/Taxol cells,which may regulate MDR1 and P-gp expression by targeting HIPK2.

Cell Transformation, Neoplastic ; drug effects ; genetics ; Dimethyl Sulfoxide ; pharmacology ; Electroporation ; methods ; HL-60 Cells ; Humans ; Membrane Proteins ; genetics ; Neoplasm Proteins ; genetics ; RNA Interference ; RNA, Small Interfering ; genetics ; Stromal Interaction Molecule 1 ; Transfection

Background Presbyopia is one of primary causes affecting the visual and life qualities of the agings,and its mechanism is associated with the oxidative damage of lens epithelial cells with ageing.SS31 is a mitochondria-targeted antioxidant peptide.To study the effect of SS31 on oxidative damage of lens epithelial cells has an important significance for the prevention and treatment of presbyopia.Objective This study was to investigate the effect of SS31 on in vitro oxidative damaged human lens epithelial cells.Methods Human lens epithelial cell line (HLEB-3) was cultured using DMEM with low glucose and 10％ fetal bovine serum(FBS).The cell model of oxidative damage was established by adding 200 μmol/L tea-butyl hydropeoxide (t-BHP) into DMEM for 18 hours.The cells were divided into blank control group,t-BHP model group,10 nmol/L SS31 +t-BHP group,100 nmol/L SS31 +t-BHP group,1 μmol/L SS31 +t-BHP group,10 μmol/L SS31 +t-BHP group and 100 pμmol/L t-BHP group,and then MTT assay was used to detect the survival rate of the cells and evaluate the optimal SS31 concentration for sequential study.The cells then were divided into blank control group,t-BHP model group and 1 μmol/L SS31 +t-BHP co-culture group.The change of mitochondrial membrane potential of the cells was tested by JC-1 dye and flow cytometry.Reactive oxygen species (ROS) level in the mitochondria was determined using MitoSOX staining.Results The cell survival rate in the t-BHP model group was (53.42±2.52)％,and that in the blank control group was 100％.The cell survival rate was considerably increased in various concentrations of SS31 groups,showing a significant difference among different groups (F=58.349,P＜0.01).A highest survival rate was (82.13 ±3.15) ％ in the 1 μmol/L SS31 +t-BHP co-culture group,which was statistically significant in comparison with the t-BHP model group (t =28.710,P＜0.05).JC-1 dye and flow cytometry assay showed that the ratio between red and green fluorescence intensity was 7.07 ±0.06 in the blank control group,4.46±0.14 in the t-BHP model group and 5.76±0.26 in the 1 μmol/L SS31 +tBHP co-culture group,showing significant difference among the 3 groups (F=172.332,P＜0.01).The ratios between red and green fluorescence intensity in the blank control group and 1 μmol/L SS31 +t-BHP co-culture group were higher than that in the t-BHP model (t =2.609,1.303,both at P＜0.001).ROS fluorescence cells were much more in the t-BHP model group compared with blank control group and 1 μmol/L SS31 + t-BHP co-culture group.Conclusions SS31 can protect HLEB-3 cells from oxidative stress.SS31 may serve as a potential new approach to the treatment of presbyopia and other age-related diseases of lens.

RNA-Sequencing (RNA-Seq) is a newly-developed method in transcriptome research, it can afford more accurate transcription information and be more quickly by using Next-generation Sequencing (NGS) technology. RNA-Seq has been widely used in various biological fields. Genuine traditional Chinese medicines (TCM), with good quality and therapeutic effect, were always praised highly and used by famous physicians. The geo-herbalism formation of TCM is based on the product of the gene expression at specific space and time. So it has been a research hotspot to analyze the mechanism of biosynthesis through RNA-Seq in the study on the secondary metabolism of medicinal plant. This article mainly illustrates the RNA-Seq and its advantages, it also discusses the potential application in genuine TCM, and it can provide useful information for other researchers.
Drugs, Chinese Herbal ; Gene Expression Profiling ; High-Throughput Nucleotide Sequencing ; Medicine, Chinese Traditional ; Plants, Medicinal ; genetics ; RNA ; Sequence Analysis, RNA ; Transcriptome

The effects of dietary supplementation with Clostridium butyricum on growth performance and humoral immune response in Miichthys miiuy were evaluated. One hundred and fifty Miichthys miiuy weighing approximately 200-260 g were divided into five groups and reared in 15 tanks with closed circuiting culture system. The animals were fed 5 diets: basal diet only (control) or supplemented of the basal diet with C. butyricum at doses of 10(3) (CB1), 10(5) (CB2), 10(7) (CB3) or 10(9) (CB4) CFU/g. Compared with the control, the serum phenoloxidase activity was significantly increased by the supplementation (P<0.05), acid phosphatases activity was increased significantly (P<0.05) at the doses of 10(9) CFU/g. Serum lysozyme activity peaked at dose of 10(7) CFU/g and in the skin mucus at dose of 10(9) CFU/g. Immunoglobulin M level in the serum and skin mucus was increased except at dose of 10(3) CFU/g (P<0.05). The growth at the dose of 10(9) CFU/g was higher than that of the control (P<0.05). It is concluded that supplementation of C. butyricum can mediate the humoral immune responses and improve the growth performance in Miichthys miiuy.
Animal Feed ; microbiology ; Animals ; Antibody Formation ; physiology ; Clostridium butyricum ; immunology ; Dietary Supplements ; microbiology ; Fishes ; growth & development ; immunology ; Probiotics ; administration & dosage