Objective:To establish an antibody expression system to reduce the antibody-dependent enhancement (ADE) effect of target antibody.Methods:Site-directed mutagenesis was used to mutate the 234 and 235 sites of the Fc region of the mammalian cell antibody expression vector-L234A and L235A to establish the antibody expression vector pFRT-IgG1κ-FcM. An antibody Wt-WNV with significant ADE effect obtained in previous work was selected and expressed by the pFRT-IgG1κ-FcM system to obtain mutant antibody FcM-WNV. The binding ability of FcM-WNV to target antigen West Nile virus envelope protein-DⅢ (WNV E-DⅢ) was detected by ELISA, and the its binding ability to human high-affinity IgG Fc receptor hFcγRⅠ (hCD64 ) was analyzed by flow cytometry. The neutralizing activity of FcM-WNV in vitro was detected by pseudovirus infection of host cells (BHK21 and K562). Results:The expression levels of FcM-WNV and Wt-WNV were comparable, and FcM-WNV could recognize and bind to WNVE-DIII in a concentration-dependent manner. Compared with Wt-WNV, the binding ability of FcM-WNV to hCD64 was significantly weakened, showing a significant decrease in fluorescence intensity. Consistent with the previous experimental results, Wt-WNV at a concentration of 5 μg/ml significantly enhanced the infection of K562 by WNV pseudovirus, while FcM-WNV at a concentration of 5 μg/ml could effectively block pseudovirus infection in both K562 and BHK21 cells.Conclusions:The established antibody expression system can effectively reduce the ADE effect of the target antibody.

Objective:To establish an in vivo infection model of West Nile virus (WNV) pseudovirus and evaluate the neutralizing activity of antibody WNV-XH1.Methods:A stable cell line that can package the WNV pseudovirus was established in the early stage to prepare the pseudovirus supernatant. The supernatant was concentrated and infected BHK21 cells to detect the titer of the pseudovirus. After intraperitoneal injection of the pseudovirus into C57BL/J mice, bioluminescence imaging was performed to observe the infection status of the pseudovirus in the mice. After simultaneous infection, blood was collected and ELISA was used to detect NS1 levels in mouse serum. The in vivo functional activity of antibody WNV-XH1 was evaluated using the established mouse infection model.Results:Fluorescence was detected in C57BL/J mice infected with WNV pseudovirus, and the NS1 levels in the peripheral blood serum of mice infected with pseudovirus were significantly higher than those of non infected mice (1.453±0.09vs0.305±0.018). After intravenous administration of WNV-XH1 antibody before the attack, the fluorescence signal in the mice decreased and the serum NS1 level decreased (0.384±0.015).Conclusions:A successful in vivo infection model of WNV pseudovirus was established, and it was confirmed that the antibody WNV-XH1 had a protective effect against WNV pseudovirus infection in vivo.

Objective:To investigate the status quo of psychological distress among parents of children with cancer and analyze its influencing factors.Methods:Totally 334 parents of pediatric oncology patients meeting the inclusion criteria were selected by convenience sampling from Hunan Children's Hospital between June and October 2022. A questionnaire survey was conducted using a general information questionnaire, Distress Thermometer, Social Support Rating Scale, and Mishel Uncertainty in Illness Scale-Family Member Form.Results:A total of 334 questionnaires were distributed, with 298 valid responses received, making the effective response rate 89.22%. The DT score of the 298 parents of pediatric oncology patients was (7.58±2.85). Multiple linear regression analysis revealed that parental gender, the impact of caregiving on income, economic pressure, uncertainty in illness, and social support were significant influencing factors of the parents' psychological distress ( P<0.05) . Conclusions:The psychological distress of parents of children with cancer is at a relatively high level and is influenced by multiple factors. Medical professionals can implement targeted interventions based on specific circumstances to alleviate the psychological distress of these parents.

The essence-qi-spirit theory is an important part of traditional Chinese medicine， whose steady state is the material and functional basis for the balance of yin and yang in the body， making the essence， qi and spirit integrated， and body and spirit harmonized. Based on this theory， it is proposed that essence and qi depletion， spirit dissipation and qi dispersion， disharmony between yin and yang is the main pathogenesis of sleep disorders. Therefore， the method of regulating and harmonzing yin and yang by essence gathering， qi nourishing and spirit storing can be used to treat sleep disorder. The biological clock system of the circadian rhythm of sleep is regulated by the molecular oscillation that is generated by the transcription of the biological clock gene， and is a clock gradually formed by orga-nisms constantly adapting to the laws of nature. As the material basis， power， and embodiment of sound and peaceful sleep， essence， qi and spirit can perceive and transmit natural signals， whose functions are similar to what is recognized by modern science that oscillation amplifies the rhythm signal， and synchronously regulates the expression signal of the biological clock gene， thereby forming a biological clock system with “input-oscillation-output” as the feedback cycle. It is believed that the regulation method of yin and yang by essence gathering， qi nourishing and spirit storing may comprehensively regulate the physiological activities through brain/ muscle aryl hydrocarbon receptor nuclear translocator-like protein 1 （BMAL1）/circadian locomotor output cycles kaput （CLOCK）-period protein （PER）/ cryptochrome （CRY） transcriptional feedback loop， thereby adapting to the natural environment changes， playing an active role in the treatment of sleep disorders， and provideing a new idea for traditional Chinese medicine to reshape the molecular regulation system of the endogenous biological clock to prevent and treat sleep disorders.

Patient experience is an important window for modern hospital management and medical service quality, and also an important point for humanistic hospital construction. Improving patient experience is of great significance for constructing harmonious doctor-patient relationship and improving patients’ satisfaction and sense of gain. Based on the perspective of patient experience improvement, this paper analyzed the core value of "patient demand first" of Mayo Clinic and the operational experience of Cleveland Medical Center Patient Experience Office, and applied them to the cultivation of humanistic hospital culture in the First Affiliated Hospital of Xi’an Jiaotong University. Based on the concept of "strengthening patient demand orientation", with the goal of "building a high-quality patient service system and significantly improving patient satisfaction", through building education platforms for medical ethics and medical humanism, exploring the medical humanism dissemination mode for medical staff, and carrying out the humanism literacy cultivation practice in the training camp for improving hospital service efficiency, the humanistic cultural atmosphere of the hospital has been further enhanced, and the patient satisfaction has been steadily improved. It provides reference for the cultivation of humanistic literacy and the construction of new culture in public hospitals in the context of high-quality development.

Objective:To conduct a meta-analysis to analyze the efficacy and adverse reactions of fractionated high dose rate brachytherapy (HDR-BT) as monotherapy for localized prostate cancer.Methods:Relevant databases were searched to collect the clinical trials on HDR-BT as monotherapy in patients with localized prostate cancer. Included studies were limited to full-text publications of fractionated HDR-BT as monotherapy with a median follow-up of at least 5 years, and adequate reporting of treatment outcomes and adverse reactions data. Stata 12.0 was used for data analysis.Results:According to the inclusion and exclusion criteria, a total of 11 clinical trials involving 2 683 patients with prostate cancer were included in this meta-analysis. The results of the meta-analysis showed that 5-year biochemical recurrence-free survival (bRFS) rate and overall survival (OS) rate were 94% (95% CI: 93% - 96%) and 96% (95% CI: 94% - 98%), respectively. Long-term (≥5 years) cancer-specific survival (CSS) rate and distant metastasis-free survival (DMFS) rate were 99% (95% CI: 98% - 100%) and 98% (95% CI: 98% - 99%), respectively. Long-term (≥5 years) late grade ≥3 grade gastrointestinal and genitourinary adverse reactions rates were 2% (95% CI: 1% - 3%) and 9% (95% CI: 6% - 13%), respectively. Conclusions:Fractionated HDR-BT as monotherapy is an effective treatment for patients with localized prostate cancer. Its long-term efficacy is encouraging, and the treatment is well tolerated and safe.

Objective:To establish an in vivo infection model of H5N1 pseudovirus and to detect the neutralizing activity of FHA3 antibody using this model. Methods:Based on the sequence information of hemagglutinin (HA) and neuraminidase (NA) of A/Anhui/1/2005/H5N1 strain, two recombinant plasmids of pcDNA3.1-HA5 and pcDNA3.1-NA1 were constructed. The two plasmids and plasmid pNL4-3.Luc.R-E- were co-transfected into 293T cells to prepare H5N1 pseudovirus supernatant. The morphology of pseudovirus particles in the supernatant was observed by electron microscopy. MDCK cells were infected with the pseudovirus supernatant and the virus titer was detected. BALB/c mice were injected with the pseudovirus supernatant by intraperitoneal injection and subjected to bioluminescence imaging at 2, 5, 8, and 12 d after infection to detect the pseudovirus infection in vivo. The functional activity of FHA3 antibody in vivo was evaluated using the established mouse infection model. Results:The recombinant plasmids pcDNA3.1-HA5 and pcDNA3.1-NA1 were correctly constructed and could be used to prepare pseudovirus supernatants of high titer by co-transfecting 293T cells with the plasmid pNL4-3.Luc.R-E-. The virus particles were round under electron microscope. H5N1 pseudovirus-infected mice exhibits strong fluorescence signals, which were attenuated by FHA3 treatment before challenge.Conclusions:The in vivo infection model of H5N1 pseudovirus was successfully constructed and FHA3 antibody was proved to be protective against the pseudovirus infection.

Objective:To learn about the application of medical ethics review in scientific research and prevention of endemic diseases.Methods:The method of retrospective analysis was used, original articles on field epidemiological investigation and clinical medicines published by Chinese Journal of Epidemiology from 2017 to 2020 were collected. Examination of medical ethics in national, provincial and municipal fund programs and nonfund projects was carried out. Statistical analysis was performed using Cochran-Armitage trend test and Cochran-Mantel-Haenszel (CMH) test.Results:A total of 638 articles were collected from 2017 - 2020, with 36 excluded and 602 remaining. The proportion of papers published after medical ethics review over the past four years was 56.85% (83/146), 62.50% (105/168), 59.87% (94/157), and 60.31% (79/131), respectively. There was no statistically significant difference in trend testing ( Z = 0.41, P > 0.05). There was no statistically significant difference in the proportion of papers produced by academic works, on-site investigations, and clinical medicine programs among different years (χ 2 = 0.01, 1.31, 1.92, P > 0.05). The proportion of papers published that supported by various fund programs that had undergone medical ethics review over the past four years was 60.55% (66/109), 62.28%(71/114), 62.38% (63/101), and 60.22% (56/93), respectively. The trend test showed no statistically significant difference( Z = - 0.03, P > 0.05). There was no statistically significant difference in the proportion of papers published that supported by national level projects, provincial and ministerial level projects, and municipal level projects among different years (χ 2 = 0.06, 0.02, 0.19, P > 0.05). The difference in the trend test of the output papers of research projects approved by the higher-level ethics committee and marked with approval numbers over the past 4 years was statistically significant ( Z = 2.85, P < 0.01); the difference was statistically significant when compared across years (χ 2 = 8.13, P < 0.01); the proportion of papers increased from 8.22% (12/146) in 2017 to 18.08% (25/131) in 2020 (χ 2 = 7.04, P = 0.008). There was no statistically significant difference ( Z = - 0.53, P > 0.05) in the proportion of papers that expressed their consent in terms of informed consent over the past 4 years; There was no statistically significant difference in comparison between different years (χ 2 = 0.28, P > 0.05). Conclusions:Medical ethics review has been taken seriously by the majority of researchers and is widely used in endemic scientific research and prevention projects.

Objective:To prepare and identify a broad-spectrum antibody FHA3 targeting influenza A virus hemagglutinin (HA).Methods:According to the single-chain antibody fragment (scFv) sequence, the heavy chain (VH) and light chain (VL) variable regions of FHA3 were amplified by PCR and a recombinant plasmid pFRT-IgG1κ-FHA3 was constructed by linking the expression vector pFRT-IgG1κ. FHA3 was expressed in the ExpiCHO system and purified by affinity purification. The binding activity of FHA3 to influenza A virus HA was detected by ELISA. The neutralizing activity of FHA3 was detected in vitro by infecting host cells with pseudovirus. Results:SDS-PAGE showed that high-purity FHA3 was obtained. FHA3 could bind to H1N1 HA, H2N2 HA, H3N2 HA, H5N1 HA, H7N9 HA and H9N2 HA in a concentration-dependent manner. FHA3 had good neutralizing activity in vitro that was it could effectively block the invasion of H5N1 and H7N9 pseudoviruses into target cells at a low concentration of 5 μg/ml and H1N1 pseudovirus at 0.012 5 μg/ml. Conclusions:A broad spectrum antibody targeting HA protein of influenza A virus with neutralizing activity in vitro was obtained.

Objective:To prepare and identify a functional antibody FNA1 targeting the neuraminidase (NA) of influenza A virus N1 subtype.Methods:According to single-chain antibody fragment (scFv) sequence, the heavy chain and light chain variable region sequences of FNA1 were synthesized, and the recombinant expression plasmid pFRT-IgG1κ-FNA1 was constructed by linking the expression vector pFRT-IgG1κ. The FNA1 antibody was expressed in ExpiCHO cells and purified using affinity purification technique. The binding ability of FNA1 to the target proteins, influenza A virus N1 subtype NA antigens, was detected by ELISA. Flow cytometry was performed to analyze the binding ability of FNA1 to the NA antigens expressed on the surface of cell membrane. The in vitro activity of FNA1 against NA was evaluated by infecting 293T cells with pseudovirus. Results:Protein electrophoresis showed that FNA1 with high purity was obtained. FNA1 specifically recognized and bound to N1 subtype NA antigens in a concentration-dependent manner. FNA1 could effectively block NA activity by binding to N1 subtype NA protein expressed on the surface of cell membrane, thus inhibiting the release of packaged pseudovirus from cell surface and further inhibiting target cell infection.Conclusions:An antibody FNA1 targeting influenza A virus N1 subtype NA with in vitro functional activity was obtained.