Treatment advice of small molecule antiviral drugs for elderly COVID-19 / 中国药理学通报

COVID-19 has been prevalent for three years. The virulence of SARS-CoV-2 is weaken as it mutates continuously. However, elderly patients, especially those with underlying diseases, are still at high risk of developing severe infections. With the continuous study of the molecular structure and pathogenic mechanism of SARS-CoV-2, antiviral drugs for COVID-19 have been successively marketed, and these anti-SARS-CoV-2 drugs can effectively reduce the severe rate and mortality of elderly patients. This article reviews the mechanism, clinical medication regimens, drug interactions and adverse reactions of five small molecule antiviral drugs currently approved for marketing in China, so as to provide advice for the clinical rational use of anti-SARS-CoV-2 in the elderly.

Effect evaluation of reproductive health program with IEC method among junior middle school students in Shanghai / 上海预防医学

[ Objective ] To evaluate the feasibility and effectiveness of a reproductive health program with information, education and communication ( IEC ) method among junior middle school students. [ Methods] Investigation was performed by the stratified cluster sampling method.Self-report questionnaires were given to junior middle school students who were randomly included from 16 classes of 8 middle schools in Shanghai.A 3-month intervention with IEC method was conducted, which included activities as issuing intervention material, hand-copying paper competition, debating discussion, knowledge contest, scene plays and so on.Participant teachers had been trained for organization of these activities. After that, a second investigation by questionnaire was made both in intervention and control groups and then effect evaluation was done on the basis of the results of the two investigations before and after intervention. [ Results] Compared with control group, knowledge scores of intervention group were improved significantly after the intervention.The general knowledge scores were increased from 42.3 to 61.3 (P<0.01), which was apparently higher than that in control group.At posttest, the intervention group students'premarital sex attitude was more conservative compared with the control group, with the scores being increased from 66.7 to 73.9 (P<0.01).The intervention group students'scores of having premarital sex and safe sex intention were increased from 77.7, 68.7 to 89.9, 87.3 (P all<0.01).There were about four fifths students who expressed that they were satisfied with the intervention content, format, time schedule, and benefited from it. [ Conclusion ] The school-based IEC method can change the premarital sexual behavior intention in some students and some others tend to have safe sex.Short-term effect of intervention is apparent, but the long-term impact of the intervention on the students still needs to be further investigated and confirmed.

Membrane cholesterol mediates the endocannabinoids-anandamide affection on HepG2 cells / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To study the effect of anandamide (AEA) on necrosis in HepG2 cells and to explore the role of AEA in progression of liver cancer.</p><p><b>METHODS</b>Localization of the fatty acid hydrolytic enzyme (FAAH), cannabinoid receptors 1(CB1) and cannabinoid receptors 2 (CB2) proteins was detected in L02 and HepG2 cells using immunofluorescence. L02 and HepG2 cells were treated with different concentrations of AEA and methyl-beta-cyclodextrin, and the rates of cells necrosis were examined by PI stain. Meanwhile, the expression levels of FAAH, CB1 and CB2 receptor proteins, as well as P38 mitogen-activated protein kinase (p-P38 MAPK) and c-Jun-NH2-terminal kinase (p-JNK) proteins, were analyzed by Western blot.</p><p><b>RESULTS</b>The FAAH, CB1 and CB2 receptor proteins were observed both in cytoplasm and on membrane in L02 and HepG2 cells. The expression level of FAAH protein was higher in HepG2 than in L02 cells. The expression level of CB1 receptor protein was very low in both L02 and HepG2 cells. The expression level of CB2 receptor protein was high in both L02 and HepG2 cells. AEA treatment induced necrosis in HepG2 cells but not in L02 cells. Methyl-beta-cyclodextrin treatment prevented necrosis in HepG2 cells (t = 3.702; 5.274; 3.503, P less than 0.05). The expression patterns of FAAH, CB1 and CB2 receptor protein in L02 and HepG2 cells were confirmed by western blot, which were consistent with the immunofluorescence results. AEA treatment increased the levels of p-P38MAPK and p-JNK proteins in a dose-dependent manner in HepG2 cells (F = 11.908; 26.054, P less than 0.05) and the increase can be partially by prevented by MCD (t = 2.801; t = 12.829, P less than 0.05).</p><p><b>CONCLUSION</b>AEA treatment induces necrosis in HepG2 cells via CB1 and CB2 receptors and lipid rafts.</p>

Impact of PI3K /Akt /mdm2 signaling pathway on the sensitivity of gastric cancer cell line SGC7901 to doxorubicin / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To explore whether PI3K/Akt/mdm2 signalling pathway affect the sensitivity of gastric cancer cell line SGC7901 cells to doxorubicin.</p><p><b>METHODS</b>Gastric cancer cell line SGC7901 cells were exposed to doxorubicin and specific PI3K inhibitor wortmannin. Cell apoptosis was detected using flow cytometry. PI3K activity was detected by radioactive immunoprecipitation-kinase assay. Western blotting was employed to evaluate the expressions of PI3K-p85, pAkt-S473, Akt, pmdm2-S166 and p53.</p><p><b>RESULTS</b>The level of apoptosis in gastric cancer SGC7901 cells treated with doxorubicin was gradually increasing. wortmannin enhanced its effects significantly. PI3K activity and the expression of pAkt-S473 increased in a time-dependent manner, pmdm2-S166, p53 were also increased wortmannin inhibited phosphorylation of mdm2 and improved the p53 expression.</p><p><b>CONCLUSION</b>PI3K/Akt/mdm2 signalling pathway can be activated by doxorubicin and suppress apoptosis by promoting phosphorylation of mdm2. PI3K inhibitor wortmannin can enhance sensitivity of gastric cancer cells to chemotherapy.</p>

Effects of guanine-quadruplexes formation induced by adriamycin on telomeric extension reaction mediated by telomerase of Tca8113 cells / 华西口腔医学杂志

<p><b>OBJECTIVE</b>To study the effects of adramycin to disturb telomeric extention reaction mediated by telomerase of Tca8113 cells by inducing oligonucleotides that contain telomeric repeats to form guanine-quadruplex (G4) structures.</p><p><b>METHODS</b>In the presence of adriamycin, d(TTAGGG)4, d(TTAGAG)4, d(TTAGGG)5 and d(TTAGGGT) were analyzed by electrophoretic mobility shift assay. The mobility of d(TTAGGG)3, d(TTAGGG)4 and d(TrAGGG)5 in native polyacrylamide electrophoresis were observed. Methylation protection experiments were performed to investigate the effects of adriamycin on methylation of guanine in d(TTAGGG)4 and d(TTAGAG)4. The traditional telomeric repeats amplification protocol (TRAP) and modified TRAP-G4 assays were, respectively, used to analyze the different characteristcs of adriamycin's inhibiting telomeric extension mediated by telomerase of Tca8113 cells.</p><p><b>RESULTS</b>At 5.00 microg/mL of adriamycin, conversion of some of linear d(TrAGGG)4 and d(TrAGGG)5to the new, high-mobility bands formed by complex with special second structures were found in the mobility shift assay. Adriamycin at 1.25 microg/mL protected the G in d(TIAGGG)4 from methylating. Adriamycin at 2.50 microg/mL or 1.25 microg/mL partially inhibited the telomeric extension lengthened by telomerase of Tca8113 cells in TRAP assay, but completely did so in TRAP-G4 assay.</p><p><b>CONCLUSION</b>Adriamycin is able to disturb telomeric extention mediated by telomerase of Tca8113 cells by inducing oligonucleotides that contain telomeric repeats to form intra-molecular G4 structures.</p>

Synthetic radiation-inducible promoter mediated CDglyTK gene in treatment of Tca8113 cells / 华西口腔医学杂志

<p><b>OBJECTIVE</b>To observe the therapeutic effect of CDglyTK gene mediated by synthetic radiation-inducible promoter in the treatment of Tca8113 cells.</p><p><b>METHODS</b>CDglyTK gene in pCEA-CDglyTK was subcloned into pcDNA3.1 (+) to construct plasmid pcDNA3.1 (+)-CDglyTK, and then the synthetic radiation-inducible promoter in pMD18 -T -E was inserted into pcDNA3.1 (+) -CDglyTK to construct plasmid pcDNA3.1 (+ )/E -CDglyTK. The recombinant plasmid was transfected into Tca8113 cells by lipofectamine, and then exposed to 3 Gy irradiation. Cytotoxicity was evaluated by MTT. The expression of CDglyTK gene was detected by RT-PCR. The apoptosis and proliferation were examined by flow cytomtery.</p><p><b>RESULTS</b>The plasmid pcDNA3.1 (+)/E-CDglyTK was constructed successfully. The comparative survival rate of Tca8113 cells was markedly decreased by induction irradiation. Up-regulation of CDglyTK expression was found in Tca8113 cells exposed to irradiation. The apoptosis index (AI) of Tca8113 cells exposed to irradiation was higher than that of Tca8113 cells without irradiation, the other way round, the proliferation index (PI) of Tca8113 cells exposed to irradiation was lower than that of Tca8113 cells without irradiation.</p><p><b>CONCLUSION</b>The synthetic radiation-inducible promoter can be served as a molecular switch to improve the expression of CDglyTK gene in Tca8113 cells, and low dose induction radiation can significantly improve the therapeutic efficiency.</p>

Effects of apoptosis of Tca8113 cells induced by adriamycin on telomerase and telomere repeat binding factor proteins / 中华口腔医学杂志

<p><b>OBJECTIVE</b>To investigate the role of telomerase and telomere repeat binding factors (TRF) in apoptosis.</p><p><b>METHODS</b>The proliferative activity of Tca8113 cells was assessed by methyl thiazolyl tetrazolium (MTT) assay. After Tca8113 cells were treated with adriamycin at 5 mg/L, apoptotic morphology was observed under microscope with Giemsa staining and apoptosis examined by flow cytometry; analysis of telomerase activity was performed by TRAP-enzyme-linked immunosorbent assay; expression and expression level of TRF proteins were detected with immunohistochemical staining and immunofluorescence label assay, respectively.</p><p><b>RESULTS</b>After Tca8113 cells were treated with adriamycin at 5 mg/L for 5 days and 7 days, the cells apoptosis was found. Telomerase activity dropped in time-dependent manner. Expression of TRF proteins appeared in nucleus of the cells. No statistical difference in expression levels of TRF was observed between the treated and untreated cells.</p><p><b>CONCLUSIONS</b>Tca8113 cells apoptosis induced by adriamycin decreased telomerase activity, but did not influence the expression level of TRF proteins.</p>

Radiation-inducible promoters-mediated cdglytk gene in the treatment of buccal carcinoma in golden hamster / 中华口腔医学杂志

<p><b>OBJECTIVE</b>To observe the therapeutic effect of CDglyTK gene mediated by radiation-inducible promoters in the treatment of buccal carcinoma in Golden Hamster.</p><p><b>METHODS</b>Animal models of buccal carcinoma in golden hamster were established by painting 0.5% dimethyl-benzanthracene. The plasmids pcDNA (+) 3.1/E-CDglyTK were transfected into tumors by lipofectamine. 24 h later, the tumors were exposed to 3 Gy irradiation. Animals were monitored at regular intervals for volume of tumors. CDglyTK mRNA was assayed by RT-PCR. Apoptosis and proliferating cell nuclear antigen were detected respectively by in situ end-labeling and immunohistochemical methods.</p><p><b>RESULTS</b>Compared with control groups, the tumor was suppressed obviously by CDglyTK gene therapy combined with 3 Gy induction radiation. The expression of CDglyTK gene could be detected by RT-PCR in the transfected tumor, and up-regulation of CDglyTK expression was found in tumor exposed to radiation (P < 0.05). There was significant difference in apoptosis index or proliferation index between tumor without irradiation and tumor with irradiation (P < 0.05).</p><p><b>CONCLUSIONS</b>The radiation-inducible promoter can be served as a molecular switch to regulate the expression of CDglyTK gene in buccal carcinoma in golden hamster, and low dose induction radiation can significantly improve the therapeutic effects.</p>