Analysis of special ehealth service for corona virus disease 2019 (COVID-19) pneumonia / 北京大学学报(医学版)

OBJECTIVE@#To analyze how governments, hospitals and information technology(IT) companies use Internet technology to provide online health services during the early stage of corona virus disease 2019 (COVID-19) epidemic in January 2020 in China, and then provide suggestions and coping strategies for the later stage and post-epidemic time.@*METHODS@#We searched for information on ehealth services related to the outbreak of COVID-19 in China. The sources of information were mainstream search engines such as Baidu and the popular interactive social platforms such as Webchat. The keywords were "Internet+pneumonia", "Internet clinic", "pneumonia online clinic" and so on. The time of information was from January 20 to February 3, 2020. The key information was extracted and encoded by two persons back-to-back. The coding information included: name of organization provider, launching time, location of provider, service items, user, health workers engaging in the service, and so on. The coded information was entered and analyzed with SPSS 24.0 and Excel.@*RESULTS@#There were totally 57 projects launched by local governments, hospitals and IT companies. Most of them were launched from January 24th to 27th, the hospital and government projects services regionally, especially in eastern provinces. In this study, 90.48% of the enterprises and 100.00% of the hospitals had online fever clinic and consultation services for COVID-19, 66.67% of the enterprises and 37.04% of the hospitals serviced derivative health problems. Only a few projects provided tele-medical consultation. There were individual projects that provided online health management for home quarantine people. Physicians were the main force of various projects. In some hospital projects, there were also nurses, pharmacists and professional technicians to provide featured consultation.@*CONCLUSION@#Ehealth is useful and helpful for the health care system to rapidly cope with health demand during instantaneous and post epidemic time. Regional distribution of ehealth is unbalanced. There are institutional and technical feasibilities for the emergency application of Internet technology. However, community health centers seldom provide ehealth or connect with tertiary hospitals with Internet. Therefore, all kinds of providers within healthcare system should promote emergence ehealth. Tele-medical diagnosis and referral should be developed by local governments during COVID-19. The application of "Internet+medical treatment" in community medical institutions and synergy among various institutions should be promoted.

ADAR1 up-regulates ZNF655 expression via RNA editing and enhances HBV replication in HepG2 cell line / 基础医学与临床

Objective To explore the effect of ADAR1 on ZNF655 and the regulation of ZNF655 on the expression of HBV. Methods Sanger sequencing was used to validate the 3′UTR region of ZNF655 in ADAR1. The expression of ADAR1 and ZNF655 mRNA as well as HBV RNA were detected by RT-qPCR. Dual luciferase report plasmid assay was used to detect the expression of luciferase. To detect the expression of ADAR1 and ZNF655 pro-tein by Western blot. HBsAg and HBeAg was detected by ELISA. Results The chr7:99575277 loci on ZNF655 3′UTR was homozygous in DNA level and hybrid in RNA level. On the 3′UTR editing site of ZNF655,the luciferase activity of the edited G allele was significantly higher than that of the normal A allele (P＜0.001). The expression of ZNF655 was upregulated by ADAR1 in the level of transcription and translation(P＜0.01).ZNF655 significantly promoted the expression of HBV. Conclusions The chr7:99575277 loci on ZNF655 3′UTR is edited by ADAR1, promoting the expression of ZNF655,which upregulated the expression of HBV.

Xuebijing Injection () increases early survival rate by alleviating pulmonary vasopermeability in rats subjected to severe burns / 中国结合医学杂志

<p><b>OBJECTIVE</b>To investigate the effects of Xuebijing Injection (, XBJ) on survival rate and pulmonary vasopermeability in a rat model of severe scald injury.</p><p><b>METHODS</b>Rats were divided into two experiments: experiment 1 was monitored for 12 h post-injury for survival analysis after severe burns; in experiment 2, rats were killed for determination of pulmonary vascular permeability and pro-inflflammatory mediators. In both experiments, rats were subject to third-degree 50% total body surface area (TBSA) burns or sham injury followed by XBJ or normal saline (NS) treatment. In addition, rat pulmonary microvascular endothelium cells (PMECs) were pretreated with either XBJ or phosphate buffer saline (PBS), and then subjected to sham serum or scald serum stimulation for 2 or 6 h, followed by transwell examination for the permeability of PMECs. Meanwhile, pro-inflflammatory mediators in PMECs culture supernatant were also investigated.</p><p><b>RESULTS</b>The average survival time in the scald+XBJ group was 582.1±21.2 min, which was signifificantly longer than that in the scald + NS group (345.8±25.4 min, P<0.01). Plasma levels of tumor necrosis factor-alpha (TNF-α), E-selectin, interleukin-6 (IL-6), vascular permeability and water content of lung tissues were signifificantly increased in animals after severe burns (P<0.01). However, administration of XBJ signifificantly decreased these levels in plasma and lung tissue. In in vitro cell experiments, XBJ markedly attenuated permeability in PMECs monolayer and reduced the levels of TNF-α, IL-6 and soluble E-selectin after stimulation with scald serum (P<0.01).</p><p><b>CONCLUSIONS</b>XBJ increases early survival rate by alleviating pulmonary vasopermeability and inhibiting pro-inflflammatory mediators in rats subjected to lethal scald injury. XBJ may be a potent drug in treatment of severe burns.</p>

Systematic reviews of ganciclovir versus acyclovir for herpes simplex virus keratitis / 国际眼科杂志(Guoji Yanke Zazhi)

To assess the efficiency and reliability safety of ganciclovir to herpes simplex virus keratitis. ●METHODS: All of the randomized controlled trials for the study of ganciclovir versus acyclovir in the treatment of herpes simplex virus keratitis were collected from Cochrane Library, EMbase, PubMed, Chinese Bio -medicine Database, China Journal Full-text Database, VlP Database and WanFang Database. Then the data were extracted and evaluated by two reviewers independently. Risk of bias assessment was evaluated by a tool recommended by Cochrane Library. Revman 5. 0 software was used for statistical analysis. ●RESULTS: Finally, 14 randomized controlled trials were included, 4820 patients totally. Subgroups were used according to the number of patients and diseased eyes as well as the difference of follow-up time. For and relapse rate, ganciclovir group was overmatch acyclovir group. There were statistical differences between the two groups [RR= 1. 22, 95%CI (1. 10-1. 36); OR = 4. 50, 95% CI (2. 02-10. 04); RR = 0. 23, 95% CI (0. 10-0. 52)]. Compared with acyclovir, ganciclovir had less side - effect. There were statistical differences between the two groups [RR = 0. 12, 95%CI (0. 03 - 0. 46)]. All of the side effects of the two groups can be relieved by themselves. ● CONCLUSlON: Current evidence suggests that the ganciclovir is more efficient and safe than acyclovir in the treatment of herpes simplex virus keratitis.

Peripheral benzodiazepine receptor agonist Ro5-4864 inhibits mitochondrial permeability transition in rat heart / 生理学报

Opening of mitochondrial permeability transition (MPT) pores leads to mitochondrial injury during oxidative stress. The peripheral benzodiazepine receptor (PBR) located at mitochondrial outer-membrane has been shown to be involved in several mitochondrial functions. In the present study, we used Ro5-4864, a PBR agonist, to test if activation of PBR could prevent MPT pore opening during Ca(2+) overloading. Cardiac mitochondria isolated from Sprague-Dawley rats were treated by 150 mmol/L Ca(2+) to induce MPT. Ro5-4864 (50, 100 and 200 micromol/L) was added into incubation buffer before adding 150 micromol/L Ca(2+). In additional group, atractyloside (ATR, 20 micromol/L), an opener of MPT pores was added 5 min before the addition of 100 micromol/L Ro5-4864. The change of absorbance at 520 nm was monitored with a spectrophotometer at 30 degrees C for 10 min. Western blot was used to detect cytochrome C loss. The mitochondrial membrane potential was monitored with the fluorescence dye JC-1. Ro5-4864 inhibited the decrease of absorbance at 520 nm compared to that in the untreated Ca(2+) group (P<0.01, P<0.05). In the presence of ATR, Ro5-4864 was not able to prevent MPT anymore. Opening of MPT pores by Ca(2+) decreased the content of cytochrome C in mitochondria, but increased cytochrome C content in cytosol. Ro5-4864 preserved cytochrome C content in mitochondria and led to less cytochrome C release to cytosol. ATR treatment reversed the protective effect of Ro5-4864 on cytochrome C content. Opening of MPT pores led to mitochondrial depolarization, whereas Ro5-4864 treatment maintained mitochondrial membrane potential. Thus, prevention of MPT by activation of PBR during calcium overloading maintains mitochondrial cytochrome C content and membrane potential. Activation of PBR during cardiac ischemia and reperfusion may be an alternative way for cardioprotection.

Preparation of Fe3O4-magnetic nanoparticles loaded with adriamycin and its reversal of multidrug resistance in vitro / 中国实验血液学杂志

To prepare Fe(3)O(4)-magnetic nanoparticles loaded with adriamycin and investigate the reversal role of drug-loaded nanoparticles in K562 and resistant cell line K562/A02, the drug-loaded nanoparticles were prepared by using mechanical absorption polymerization at different conditions of 4 degrees C or 37 degrees C for 24 or 48 hours. The survival of cells cultured with drug-loaded nanoparticles for 48 hours was detected by MTT assay, then the growth inhibition efficacy of cells was calculated. The results showed that the growth inhibition efficacy of both two cell lines was enhanced with increasing concentration of Fe(3)O(4)-magnetic nanoparticles. The inhibitory ratio of two cell lines obtained at 4 degrees C and for 48 hours was significantly better than that at 37 degrees C and 24 hours. In conclusion, Fe(3)O(4)-magnetic nanoparticles can load adriamycin by using mechanical absorption polymerization, but depended on proper temperature and time. Furthermore, drug-loaded nanoparticles showed an ability reversing multidrug resistance.

Sialographic, sialoendoscopic and irrigation fluid study in chronic obstructive parotitis / 中华口腔医学杂志

<p><b>OBJECTIVE</b>To study the sialographic changes and to compare the changes with sialoendoscopic and irrigation fluid findings in chronic obstructive parotitis (COP).</p><p><b>METHODS</b>This study involved 27 patients with a long history of parotid swelling. All patients were examined by X-ray, sialography, and were diagnosed as COP without sialolithiasis. Sialoendoscopy was used to observe the ductal system and irrigation treatment performed. The irrigated liquid was centrifuged and the deposits of fluid were stained and observed under microscopy. The sialographic changes were classified as previous studies and compared with sialoendoscopic and irrigation fluid findings.</p><p><b>RESULTS</b>The sialographic changes of COP in 27 patients included 9 cases with type I, 5 cases with type II, 9 cases with type III and 3 cases with type IV changes, 1 case was normal. Marked obstructive factors such as stricture of ductal system were revealed in 21 cases on the sialogram. Sialoendoscopic examination showed that the ductal system was filled with fiber-like substances and hyperaemia of ductal wall in all cases. While few and thin fiber-like substances were found in the COP with sialographic type I and type II changes, many thick wadding or mass fiber-like substances were revealed in COP with sialographic type III and IV changes. Microstones were found in 2 COP with sialographic type III changes which were stained and identified by microscopy. Foreign body (drug bar) was found in one COP with sialographic type I changes with sialoendoscopy. Irrigation fluid examination showed fiber-like substance was composed of desquamative duct epithelial cells, neutrophil, lymphocytes, acidophile. Some epithelial cells were found in two microliths.</p><p><b>CONCLUSIONS</b>The pathological basis of fiber-like substance on sialoendoscopy is desquamative duct epithelial cells. Fiber-like substance in the lumen of ductal system is considered as one of the obstructive factors in COP. Sialoendoscopic findings is related to sialographic changes.</p>

Study on the expression and mutation of human telomeric repeat binding factor (hTRF1) in 10 malignant hematopoietic cell lines / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

<p><b>OBJECTIVE</b>Detecting the expression and mutation of human telomeric repeat binding factor (hTRF1) in 10 malignant hematopoietic cell line cells on the base of determining its genomic structure and its four pseudogenes to clarify if hTRF1 mutation is one of the factors of the activation of telomerase.</p><p><b>METHODS</b>hTRF1cDNA sequences were obtained from GenBank, its genome structure and pseudogenes were forecasted by BLAST and other biology information programs and then testified by sequencing. Real-time RT-PCR was used to detect the expression of hTRF1mRNA in 10 cell line cells, including myelogenous leukemia cell lines K562, HL-60, U-937, NB4, THP-1, HEL and Dami; lymphoblastic leukemia cell lines 6T-CEM, Jurkat and Raji. Telomerase activities of cells were detected by using telomeric repeat amplification (TRAP)-ELISA protocol. PCR and sequencing were used to detect mutation of each exon of hTRF1 in 10 cell line cells.</p><p><b>RESULTS</b>hTRF1 gene, mapped to 8q13, was divided into 10 exons and spans 38.6 kb. Four processed pseudogenes of hTRF1 located on chromosome 13, 18, 21 and X respectively, was named as PsihTRF1-13, PsihTRF1-18, PsihTRF1-21 and PsihTRF1-X respectively. All cell line cells showed positive telomerase activity. The expression of hTRF1 was significantly lower in malignant hematopoietic cell lines cells (0.0338, 0.0108-0.0749) than in normal mononuclear cells (0.0493, 0.0369-0.128) (P=0.004). But no significant mutation was found in all exons of hTRF1 in 10 cell line cells. Four variants were found in part of intron 1, 2 and 8 of hTRF1. Their infection on gene function is unknown and needs further studies.</p><p><b>CONCLUSION</b>hTRF1 mutation is probably not one of the main factors for telomerase activation in malignant hematopoietic disease.</p>

Localization of human telomere repeat binding factor 1 in telomerase-positive and-negative cells and its expression during cell cycle / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To observe the distribution pattern of human telomere repeat binding factor 1(TRF1) in the telomerase-positive (HeLa) and telomerase-negative cells (WI38-2RA) and to investigate its expression level during the cell cycle.</p><p><b>METHODS</b>The full-length sequences of TRF1(TRF1FL) and its mutant with N and C terminus deletion (TRF1DeltaNC) were generated by PCR amplification, the resulting fragments were cloned into pEGFP-C2 mammalian expression vector. GFP-tagged proteins were verified by Western blotting with rabbit anti-TRF1 and mouse anti-GFP antibodies after cell transfection. Immunofluorescence staining were performed to detect the TRF1 localization in HeLa and WI38-2RA cells. Metaphase spreads from HeLa cells were also prepared to observe TRF1 localization in chromosomes. HeLa cells were arrested by thymidine and nocodazole at different cell stages. Cell cycles were analyzed by flow cytometry and TRF1 levels were evaluated by semi-quantitative Western blotting.</p><p><b>RESULTS</b>TRF1FL and TRF1PNC fragments were sized about 1.3 kb and 0.95 kb. GFP-tagged TRF1FL and TRF1DeltaNC proteins were 80 kD and 60 kD, respectively. In both HeLa and WI38-2RA cells, TRF1FL had a speckled distribution in the nuclei,however, TRF1FL did not coincide with promyelocytic leukemia (PML) nuclear body in HeLa cells while it exclusively did in WI38-2RA cells. Moreover, TRF1FL was exactly localized at the termini of metaphase spreads in HeLa cells. In contrast, TRF1PNC was diffusely distributed throughout the nuclei. Analysis by semi-quantitative Western blotting indicated that TRF1 levels increased with cell cycle progression, which reached the zenith at the M phase and went down to the nadir at G1/S point. The TRF1 level at M phase was about 3.9 times than that at G1/S point(t=12.92iP<0.01).</p><p><b>CONCLUSION</b>TRF1 has a different localization in telomerase-positive and telomerase-negative cells, which suggests TRF1 might exert different functions in these cells. TRF1 level is regulated with cell cycle.</p>

Telomerase activity of human bone marrow mesenchymal stem cells / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To investigate the telomerase activity in mesenchymal stem cells (hMSCs) from human bone marrow after their in vitro committed differentiation into adipocytes and cryopreservation.</p><p><b>METHODS</b>hMSCs were isolated from human bone marrow. The isolated hMSCs were induced to differentiate into adipocytes in vitro or cryopreserved. TRAP assay (telomerase repeat amplification protocol assay) was employed to detect telomerase activity in those hMSCs.</p><p><b>RESULTS</b>Telomerase activity (RTA) in hMSCs (n=19) was (1.46 +/-0.67)%, while that in hMSCs-derived adipocytes (n=3) was (11.80 +/-2.52)% (P<0.001). RTA of hMSCs-passage 1.3 (n=10) was (1.46+/-0.83)%, and that of hMSCs-passage 4-7(n=9) was (1.46 +/-0.47)% (P=0.99). Cryopreservation did not affect the telomerase activity in hMSCs, RTA of fresh hMSCs (n=13) was (1.41 +/-0.44)%, RTA of frozen hMSCs (n=6) was (1.57 +/-1.07)% (P=0.64).</p><p><b>CONCLUSION</b>hMSCs are telomerase-negative, but telomerase activity in hMSCs-derived adipocytes is upregulated.</p>

Isolation of Tara protein and its gene cloning / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To isolate and identify TRF1 immunoprecipitating protein complex and to clone the candidate gene.</p><p><b>METHODS</b>The co-immunoprecipitation assay was employed to isolate TRF1 protein complex and the immunoprecipitate was subjected to MALDI-TOF mass spectrometry for protein identification. The candidate gene was amplified by temperature-gradient PCR from human testis cDNA library and then cloned into pEGFP-C2 vector for eukaryotic expression. The amplified gene was verified by direct sequencing and GFP-tagged protein was confirmed by immunoblotting.</p><p><b>RESULTS</b>Tara protein with the size of 68 kD was identified from the TRF1 precipitate. The candidate gene amplified from cDNA library was about 1.7 kb as expected. Sequencing demonstrated the amplified fragment had 99.9% of homogenesis with Tara CDS sequence (gi:30474869). GFP-tagged fusion protein was about 100 kD. Tara was diffusely distributed in cytoplasm at interphase and in whole cells at mitotic phase.</p><p><b>CONCLUSION</b>Tara might be an interacting protein with TRF1. However, further investigation would be required to confirm if they were bona fide partners.</p>

Expression of human telomere repeat binding factor 1 (TRF1) in acute leukemia cells and its correlation with telomerase activities / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To study the expression of human telomere repeat binding factor 1 (TRF1) to investigate the correlation of telomerase activity with acute leukemia.</p><p><b>METHODS</b>Leukemic cells were collected from 30 cases of acute leukemia. Realtime quantitative PCR with fluorescence probe hybridization was used to measure expression of TRF1 and hTERT mRNA in leukemic cells.</p><p><b>RESULTS</b>TRF1 mRNA expression was 0.0126 (0.0127-0.0546) in acute non-lymphocytic leukemia (ANLL), which was lower than that in normal mononuclear cells [0.0457 (0.00839-0.262), P<0.001], but its expression in acute lymphoblastic leukemia (ALL) cells [0.0745 (1.92 x 10(-6)-0.193)] had no significant difference compared with that in normal mononuclear cells. TRF1 expression in ANLL cells was significantly lower than that in ALL cells (P=0.001). The expressions of TRF1 mRNA in AL cells and normal mononuclear cells had no significant correlation with expression of hTERT mRNA (r=-0.173, P=0.207).</p><p><b>CONCLUSION</b>The expression of TRF1 is lower in ANLL cells, which indicates TRF1 may have some effect on telomerase activity by regulating telomere length in ANLL cells.</p>

Expression of Pin1 in malignant hematopoietic cells and its relation with cell cycle / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To study the expression of peptidyl-prolyl cis/trans isomerase (PPIase or Pin1) in malignant hematopoietic cells and its relation with cell cycle.</p><p><b>METHODS</b>Realtime quantitative PCR with fluorescence probe hybridization was used to measure expression of Pin1 mRNA in malignant hematopoietic cell lines and normal mononuclear cells separated from bone marrow. HeLa cells were blocked with Thymidine and Nocodazole in different cell phases and then the expression of Pin1 mRNA and protein were detected by realtime-PCR and immunoblotting.</p><p><b>RESULTS</b>The expression of Pin1 in malignant hematopoietic cell lines was significantly higher than that in normal controls (0.339 +/-0.093 compared with 0.038 +/-0.005, P<0.01). Its expression in myeloid malignant hematopoietic cell lines was significantly higher than that in normal controls (0.388 +/-0.115 compared with 0.038 +/-0.005, P<0.01) and so was the malignant lymphocytic cell lines (0.226 +/-0.166 compared with 0.038 +/-0.005, P<0.01). The expression of Pin1 was closely correlated with cell cycle. It was the highest in G1 phase and the lowest in S phase (110.762 +/-16.737 compared with 4.080 +/-0.634, P<0.01).</p><p><b>CONCLUSION</b>Pin1 is overexpressed in malignant hematopoietic cell lines and its expression is different during cell cycle that is highest in G1 phase and lowest in S phase.</p>