Hydrophobicity of the C-terminal GPI-anchor Attachment Signals Determines ER-associated Degradation Pathway of Precursor Proteins / 中国生物化学与分子生物学报

More than 150 glycosylphosphatidylinositol (GPI)-anchored proteins (GPI-APs) are expressed in mammalian cells and involved in various physiological processes such as immune recognition, cell communication and signal transduction. GPI is transferred to proteins in the endoplasmic reticulum (ER). When GPI-anchoring is impaired, precursor proteins are thought to be degraded through ER-associated degradation (ERAD). However, the mechanism of their degradation in ERAD remains unclear. To investigate the impact of ERAD pathways on degradation of GPI precursor proteins, we used series of knockout (KO) human embryonic kidney 293 (HEK293) cells defective in PIGS gene, which encodes a GPI transamidase complex subunit, combined with KO in HRD1 (PIGS-HRD1-KO) or GP78 (PIGSGP78-KO), which encodes the E3 ubiquitin ligases for the ERAD pathways. We compared the stability of 16 GPI precursor proteins in the ERAD-deficient cells with the parental PIGS-KO cells. Western blotting data showed that the GPI precursor proteins were stabilized in either PIGS-HRD1-KO (I

Aerosol transmission of coronavirus in hospital / 上海预防医学

Coronavirus is human-to-human transmissible and often leads to nosocomial outbreak in the early stage, which poses a great threat to healthcare workers.Due to confined space, crowed population and some aerosol-generating medical procedures, hospital is vulnerable to coronavirus transmission and nosocomial infection.We review the mechanism and risk factors of coronavirus aerosol in hospital and summarize the evidence of aerosol transmission of three coronaviruses and the impact of air flow.Furthermore, we evaluate the effectiveness of personal protection in the above-mentioned scenarios for the prevention and control of nosocomial infection.

Status of microbiological reports at partial provincial level annual academic conference of healthcare-associated infection from 2015 to 2017 / 中国感染控制杂志

Objective To understand the present status of special reports relating to microbiology at China provincial-level annual academic conference of healthcare-associated infection (HAI), and provide basis for the training and capacity-building of HAI management staff.Methods The conference arrangements for annual academic conference of provincial HAI quality control centers and HAI management societies from January 2015 to May 2017 were collected, and the special reports on microbiology were summarized and analyzed.Results A total of 35 annual academic conferences held by 17 provinces/municipalities directly under the central government/autonomous regions were included in the study, most conferences (42.31％) were concentrated on the fourth quarter. 15 annual academic conferences were related to microbiology report, accounting for 5.91％ of the total reports, reporting time accounted for 4.81％ of total time. There were various forms of microbiological thematic reports, subject reports, literature exchange, and interactive exchange accounted for 68.96％, 24.14％, and 6.90％ respectively. The proportion of topics related to microbiology reports increased to a certain extent, but the proportion of reporting time decreased.Conclusion At present, the proportion and reporting time of microbiology reports in China provincial-level annual academic conference of HAI is relatively low, it is necessary to increase the number of microbiology reports in the future annual conference of HAI.

Effect of hTERT antisense oligodeoxynucleotide on telomerase activity in bladder cancer cells in vitro / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the effect of hTERT antisense oligodeoxynucleotide on telomerase activity in bladder cancer cells.</p><p><b>METHODS</b>Antisense phosphorothioate oligodeoxynucleotide (AS PS-ODN) was synthesized and purified. Telomerase activity was measured by telomerase PCR ELISA kit. hTERT mRNA expression was detected by reverse transcription-polymerase chain reaction (RT-PCR), and hTERT protein by immunohistochemistry and flow cytometry.</p><p><b>RESULTS</b>Telomerase activity was decreased in T24 cells 48 h after treatment with AS PS-ODN, and was significantly inhibited at 72 h.</p><p><b>CONCLUSION</b>AS PS-ODN can significantly inhibit telomerase activity by down-regulating hTERT mRNA and protein expression in bladder cancer cells.</p>

Comparisons of conventional and normalized calculation of quantitative real-time RT-PCR detecting PML-RAR alpha fusion gene in patients with acute promyelocytic leukemia / 中华血液学杂志

<p><b>OBJECTIVE</b>To optimize the calculation of quantitative real time RT-PCR (Q-RT-PCR) of PML-RARalpha in patients with acute promyelocytic leukemia (APL) for molecular monitoring of minimal residual disease (MRD).</p><p><b>METHODS</b>By using both regular reverse transcription polymerase chain reaction (RT-PCR) and Q-RT-PCR, the expression levels of PML-RARalpha transcripts were measured before and after treatment. The conventional Q-RT-PCR calculation was directly compared the post-treatment transcript level with the respective pre-treatment one (DoseN) in the individual patient while the standardized calculation was based on the calculation of standardized pre-treatment DoseN of all patients.</p><p><b>RESULTS</b>In 181 samples from 31 patients, the results of log-reduction of PML-RARa after induction, at the end of consolidation and during maintenance by conventional method were (1.9 +/- 1.9), (4.8 +/- 1.3) and (5.7 +/- 0.4), respectively, while by standardized method were (2.0 +/- 1.9), (4.9 +/- 1.4) and (5.7 +/- 0.1), respectively. Of notice, the result was with significant less variation of the latter methods during maintenance therapy. Moreover, with defined criteria of molecular response (3.0-4.9 log-reduction as minor and > or = 5.0 log-reduction as major molecular response), the standardized method was validated in clinical settings.</p><p><b>CONCLUSION</b>The standardized method is superior to the conventional method for calculation of Q-RT-PCR results. The new method can reduce the individual variation in monitoring the MRD and is feasible even for patients with unavailable pre-treatment samples.</p>

Inhibition of telomerase with hTERT antisense enhances TNF-alpha-induced apoptosis in prostate cancer cells PC3 / 中华男科学杂志

<p><b>OBJECTIVE</b>To investigate the inhibiting effect of telomerase with hTERT antisense on TNF-alpha-induced apoptosis in prostate cancer cells PC3.</p><p><b>METHODS</b>Antisense phosphorothioate oligodeoxynucleotide (AS PS-ODN) was synthesized and purified. Telomerase activity was measured by telomeric repeat amplification protocol (TRAP) and telomerase PCR-ELISA Kit, cell viability was determined by MTT assay, and cell apoptosis was observed by morphological method and determined by flow cytometry.</p><p><b>RESULTS</b>AS PS-ODN could significantly inhibit the telomerase activity and increase the susceptibility of TNF-alpha-induced apoptosis of PC3 cells.</p><p><b>CONCLUSION</b>Inhibition of telomerase with hTERT antisense can enhance TNF-alpha-induced apoptosis in prostate cancer cells.</p>

Inhibition of telomerase with human telomerase reverse transcriptase antisense enhances tumor necrosis factor-alpha-induced apoptosis in bladder cancer cells / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Telomerase activity is found in 85%-90% of all human cancers but not in their adjacent normal cells. Human telomerase reverse transcriptase (hTERT) is an essential component in the telomerase complex that plays an important role in telomerase activity. This study investigated the effect of the telomerase inhibition with an hTERT antisense oligodeoxynucleotide (ODN) in bladder cancer cells (T24) on tumor necrosis factor-alpha (TNF-alpha)-induced apoptosis.</p><p><b>METHODS</b>Antisense phosphorothioate oligodeoxynucleotide (AS PS-ODN) was synthesized and purified. Telomerase activity was measured by polymerase chain reaction enzyme-linked immunoassay (PCR-ELISA). hTERT mRNA expression was measured by reverse transcription polymerase chain reaction (RT-PCR) assay and a gel-image system. hTERT protein was detected by immunochemistry and flow cytometry. Cell viability was measured by the 3-(4, 5-dimethylthiazol-2-yl)-2, 5-Diphenyltetrazolium (MTT) assay. Cell apoptosis was observed by a morphological method and determined by flow cytometry.</p><p><b>RESULTS</b>AS PS-ODN significantly inhibited telomerase activity and decreased the levels of hTERT mRNA which preceded the decline in the telomerase activity. AS PS-ODN significantly reduced the percentage of positive cells expressing hTERT protein following the decline of hTERT mRNA levels. There was no difference seen in the telomerase activity, hTERT mRNA expression or the protein levels between the sense phosphorothioate oligodeoxynucleotide (SPS-ODN) and the control group. AS PS-ODN treatment significantly decreased the cell viability and enhanced the apoptotic rate of T24 cells in response to TNF-alpha while there was no difference in cell viability and apoptotic rate between the S PS-ODN and the control group.</p><p><b>CONCLUSIONS</b>AS PS-ODN can significantly inhibit telomerase activity by downregulating the hTERT mRNA and protein expression. Treatment with AS PS-ODN may be a potential and most promising strategy for bladder cancer with telomerase activity.</p>

Inhibition of telomerase with human telomerase reverse transcriptase antisense increases the sensitivity of tumor necrosis factor-alpha-induced apoptosis in prostate cancer cells / 亚洲男科学杂志(英文版)

<p><b>AIM</b>To investigate the effect of inhibition of telomerase with human telomerase reverse transcriptase (hTERT) antisense on tumor necrosis factor-alpha (TNF-alpha)-induced apoptosis in prostate cancer cells (PC3).</p><p><b>METHODS</b>Antisense phosphorothioate oligodeoxynucleotide (AS PS-ODN) was synthesized and purified. Telomerase activity was measured using the telomeric repeat amplification protocol (TRAP) and polymerase chain reaction enzyme-linked immunoassay (PCR-ELISA). hTERT mRNA was measured by reverse transcription PCR (RT-PCR) assay and gel-image system. hTERT protein was detected by immunochemistry and flow cytometry. Cell viability was detected by 3-(4,5-dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium (MTT) assay. Cell apoptosis was observed by morphological method and determined by flow cytometry.</p><p><b>RESULTS</b>The telomerase activity decreased with time after hTERT AS PS-ODN treatment. The levels of hTERT mRNA decreased with time after hTERT AS PS-ODN treatment, which appeared before the decline of the telomerase activity. The percentage of positive cells of hTERT protein declined with time after hTERT AS PS-ODN treatment, which appeared after the decline of hTERT mRNA. There was no difference in telomerase activity, hTERT mRNA and protein levels between hTERT sense phosphorothioate oligodeoxynucleotide (S PS-ODN) and the control group. The cell viability decreased with time after hTERT AS PS-ODN combined with TNF-alphatreatment. The percentage of apoptosis increased with time after hTERT AS PS-ODN combined with TNF-alpha treatment. There was no difference in cell viability and the percentage of apoptosis between hTERT S PS-ODN and the control group.</p><p><b>CONCLUSION</b>hTERT AS PS-ODN can significantly inhibit telomerase activity by downregulating the hTERT mRNA and protein expression, and inhibition of telomerase with hTERT antisense can enhance TNF-alpha-induced apoptosis of PC3 cells.</p>