Objective To explore the experiences of family caregivers in dealing with the care-resistance behaviors in elderly patients with dementia,to improve the quality of care for elderly patients with dementia,and to enhance the ability of family caregivers to cope with resistance behaviors.Methods Utilizing purposeful sampling,we conducted in-depth interviews with 16 family caregivers of elderly patients with dementia between February 2024 and May 2024.The participants were recruited from a tertiary-grade hospital and the surrounding community in Zhengzhou,Henan Province.The interview data were subjected to thematic analysis using the Colaizzi 7-step method.Results A total of 19 interviews were conducted in this study,with each caregiver being interviewed for 30-44 minutes.The total transcription was about 82,000 words,and 4 themes and 11 sub-themes were identified:the considerable physical and psychological burden(persistent negative emotions,gradually increasing physical burden),realization of self-worth(sense of filial responsibility,care benefit feeling),subjective adjustment of coping strategies(actively seeking external support,resorting to restrictions and protection,passively coping out of helplessness),and facing multiple coping challenges(difficulty in establishing effective communication,poor sleep quality,lack of caregiving skills,unmet diverse support needs).Conclusion Family caregivers experience a heavier burden of care and develop various negative emotions when dealing with resistance behaviors.Different caregivers gradually adapt and develop different coping strategies in the care process.However,there are still some coping dilemmas that need to be resolved.Healthcare professionals should pay timely attention to caregivers'experiences in dealing with resistance behaviors,correct their attitudes towards care-resistance behaviors,strengthen health education on managing care-resistance behaviors,and improve their ability to cope with care-resistance behaviors.

Objective To explore the experiences of family caregivers in dealing with the care-resistance behaviors in elderly patients with dementia,to improve the quality of care for elderly patients with dementia,and to enhance the ability of family caregivers to cope with resistance behaviors.Methods Utilizing purposeful sampling,we conducted in-depth interviews with 16 family caregivers of elderly patients with dementia between February 2024 and May 2024.The participants were recruited from a tertiary-grade hospital and the surrounding community in Zhengzhou,Henan Province.The interview data were subjected to thematic analysis using the Colaizzi 7-step method.Results A total of 19 interviews were conducted in this study,with each caregiver being interviewed for 30-44 minutes.The total transcription was about 82,000 words,and 4 themes and 11 sub-themes were identified:the considerable physical and psychological burden(persistent negative emotions,gradually increasing physical burden),realization of self-worth(sense of filial responsibility,care benefit feeling),subjective adjustment of coping strategies(actively seeking external support,resorting to restrictions and protection,passively coping out of helplessness),and facing multiple coping challenges(difficulty in establishing effective communication,poor sleep quality,lack of caregiving skills,unmet diverse support needs).Conclusion Family caregivers experience a heavier burden of care and develop various negative emotions when dealing with resistance behaviors.Different caregivers gradually adapt and develop different coping strategies in the care process.However,there are still some coping dilemmas that need to be resolved.Healthcare professionals should pay timely attention to caregivers'experiences in dealing with resistance behaviors,correct their attitudes towards care-resistance behaviors,strengthen health education on managing care-resistance behaviors,and improve their ability to cope with care-resistance behaviors.

Renal interstitial fibrosis (RIF) is the crucial pathway in chronic kidney disease (CKD) leading to the end-stage renal failure. However, the underlying mechanism of Shen Qi Wan (SQW) on RIF is not fully understood. In the current study, we investigated the role of Aquaporin 1 (AQP1) in SQW on tubular epithelial-to-mesenchymal transition (EMT). A RIF mouse model induced by adenine and a TGF-β1-stimulated HK-2 cell model were etablished to explore the involvement of AQP 1 in the protective effect of SQW on EMT in vitro and in vivo. Subsequently, the molecular mechanism of SQW on EMT was explored in HK-2 cells with AQP1 knockdown. The results indicated that SQW alleviated kidney injury and renal collagen deposition in the kidneys of mice induced by adenine, increased the protein expression of E-cadherin and AQP1 expression, and decreased the expression of vimentin and α-smooth muscle actin (α-SMA). Similarly, treatmement with SQW-containing serum significantly halted EMT process in TGF-β1 stimulated HK-2 cells. The expression of snail and slug was significantly upregulated in HK-2 cells after knockdown of AQP1. AQP1 knockdown also increased the mRNA expression of vimentin and α-SMA, and decreased the expression of E-cadherin. The protein expression of vimentin increased, while the expression of E-cadherin and CK-18 significantly decreased after AQP1 knockdown in HK-2 cells. These results revealed that AQP1 knockdown promoted EMT. Furthermore, AQP1 knockdown abolished the protective effect of SQW-containing serum on EMT in HK-2 cells. In sum, SQW attentuates EMT process in RIF through upregulation of the expression of AQP1.
Drugs, Chinese Herbal/pharmacology* ; Humans ; Animals ; Mice ; Male ; Cell Line ; Rats ; Kidney/physiology* ; Fibrosis/drug therapy* ; Renal Insufficiency, Chronic/drug therapy* ; Adenine ; Epithelial-Mesenchymal Transition ; Aquaporin 1/metabolism*

Objective:To investigate the immune characteristics of SARS-CoV-2 membrane (M) protein, especially the possibility of inducing antibody-dependent enhancement effect (ADE).Methods:Full-length SARS-CoV-2 M protein was prepared by prokaryotic expression system and purified. BALB/c mice were immunized subcutaneously three times (on day 1, day 14 and day 21) by purified M protein. Serum samples were collected before immunization and after each immunization. The specificity of immune sera against M protein was identified by Western blot, and the antibody titers were detected by ELISA and neutralization test. In the presence of anti-M protein serum, the proliferation of SARS-CoV-2 in dendritic cells, nature killer cells, T and B cells was detected in vitro. Results:The immune sera from BALB/c mice immunized with purified full-length M protein of SARS-CoV-2 specifically recognized viral M protein. The titer of anti-whole virus antibody in immune sera was about 1∶400, but the antibody could not neutralize live virus. Moreover, the antibody could not help the virus to infect and proliferate in the various types of immune cells with Fc receptor (FcR).Conclusions:Non-neutralizing antibody induced by M protein could not cause ADE through FcR pathway.

Objective:To analyze the characteristics of immune cross-reaction between herpes simplex virus type 1 (HSV-1) and HSV-2 in terms of serology and clinical protection aiming to provide data for the control and prevention of diseases caused by the two viruses.Methods:An attenuated HSV-1 strain M3 was used to immunize BALB/c mice. Specific immune responses indicated by the production of neutralizing antibodies were detected. Wild-type HSV-1 and HSV-2 strains were respectively used to infect the mice through different ways 28 d after the immunization to observe the protective immunity in the M3-immunized mice against HSV-1/2 infection.Results:M3 strain could not induce specific neutralizing antibodies against HSV-2. Therefore, viral loads in tissues of the immunized mice increased significantly following different modes of HSV-2 exposure. However, no obvious abnormal clinical manifestations were found and the histopathological damage was only slight inflammatory reaction. In contrast, HSV-1-specific neutralizing antibodies were elicited in the M3-immunizaed mice with significant protective effects against HSV-1 infection.Conclusions:The immune response induced by attenuated HSV-1 strain M3 in mice exhibited immune-protective effects characterized by production of neutralizing antibodies and inhibition of virus proliferation in vivo against wild-type HSV-1 infection. For HSV-2, instead of neutralizing virus in form of antibodies, it featured by more of clinical cross-immunoprotective abilities to control virus growth.

Objective: To prepare monoclonal antibodies against pneumonia serotype 33F polysaccharides (Pn33Fps) and hepatitis B virus (HBV) surface proteins (HBs) by using the conjugate of Pn33Fps and HBs as antigen. Methods: The conjugate of Pn33Fps and HBs was used as antigen to immunize mice with different immune doses, different immune procedures and different immune sites. Mouse spleen cells with higher antibody level after immunization were isolated and fused with SP2/0 myeloma cells. The hybridoma cells were screened specifically with Pn33Fps or HBs to prepare corresponding monoclonal antibodies. Results: Serum antibodies against Pn33Fps and HBs were induced by immunizing mice with the conjugate. Monoclonal cell lines capable of continuously expressing antibodies against Pn33Fps or HBs were obtained. It has been proved that the recovery rates of samples of Pn33Fps and HBs prepared in three batches tested with ascites monoclonal antibodies prepared by these two monoclonal cell lines were between 95% and 105%. Conclusions Monoclonal antibodies against Pn33Fps and HBs could be prepared simultaneously by immunizing mice with the conjugate of Pn33Fps and HBs and used for the quantitative detection of Pn33Fps and HBs.

Objective To investigate the molecular characteristics of immune response signaling molecules induced by transfection of coxsackievirus B2 ( CVB2 ) structural proteins into epithelial cells. Methods Recombinant eukaryotic expression plasmids containing the coding regions of CVB2 structural proteins VP1-VP4 were constructed and then transfected into 16HBE cells. Culture supernatants and cell ly-sates of the transfected 16HBE cells were collected. Expression of signaling molecules involved in innate im-mune responses in transfected 16HBE cells at mRNA level was detected by RT-Q-PCR. The proliferation of T cells co-cultured with culture supernatants and cell lysates of the transfected 16HBE cells was analyzed by ELISPOT. Results Expression of innate immunity-related signaling molecules such as TGF-β-activated ki-nase ( TAK) , NF-κB-inducing kinase ( NIK) , IκB kinase α ( IKKα) and IFN-β at mRNA level was up-regulated in 16HBE cells transfected with CVB2 structural proteins VP1-VP4. Both culture supernatants and cell lysates of the transfected 16HBE cells enhanced the proliferation of T cells. Conclusions CVB2 struc-tural proteins VP1-VP4 could enhance the expression of innate immunity-related signaling molecules to var-ying degrees and promote the activation of adaptive immunity.

Objective To observe and analyze the pathological changes in BALB/c mice infected with herpes simplex virus typeⅡ (HSV-2) through nasal and genital inoculation. Methods Six-week old female BALB/c mice were divided into two groups, experimental and control groups. In the experimental group, the mice were infected with HSV-2 (104 CCID50/20μl per mouse) through nasal and genital tract in-oculation. Accordingly, the mice in the control group were injected with equal volume of PBS. Tissue speci-mens were collected from lung, nervous system and reproductive system for pathological analysis and viral load detection at different time points after infection. Lat gene expression in mouse trigeminal and sacral gan-glia was detected through in situ hybridization. In addition, the proliferation of viruses isolated form trigemi-nal and sacral ganglia of the infected mice was observed in vitro. Results Weight loss and histopathological lesions were observed in the mice of the experimental group 6 d after infection. Major pathological changes in the HSV-2-infected mice through nasal tract inoculation involved the lung and central nervous system( CNS) , including alveolar wall congestion, cerebrovascular cuff response and lymphocyte infiltration. How-ever, the major lesions in the infected mice through genital tract inoculation were found in the reproductive ducts, such as sacral ganglion necrosis, eosinophilia in the vagina and uterus, and ovarian congestion. Re-sults of the viral load detection in tissues and organs of the infected mice were consistent with the pathological changes. The mice infected through nasal tract inoculation had significantly higher viral loads in the nerves and lungs than those by genital tract inoculation, but lower viral loads in the genital tracts and sacral ganglia. Positive expression of lat gene at mRNA level was detected in the trigeminal and sacral ganglia of mice with HSV-2 latency 28 d after infection. In addition, both of the tissue fragments from trigeminal and sacral ganglia had cytopathic effects ( CPEs) on Vero cells. Enhanced expression of lat gene at mRNA level and much severer CPEs were induced by genital tract inoculation than by nasal tract inoculation. Conclu-sions HSV-2 could infect and cause histopathological damages in BALB/c mice through both nasal and genital tracts. In addition, the locations of the pathological lesions were closely related to the mode of infection.

Objective To investigate the influences of herpes simplex virus 1 and 2 ( HSV1 and HSV2) infection on the expression of signaling molecules associated with innate immune response in respira-tory and vaginal epithelial cells for bettering understanding of HSV infection and pathological characteristics in the primary infection site, namely mucosal epithelial tissues. Methods KMB17 and VK2 cells were in-fected with HSV. Changes in cell morphology and inner structure after HSV infection were observed under optical microscope and scanning electron microscope, respectively. Viral proliferation in KMB17 and VK2 cells was detected by plaque assay, microcytopathic assay and real-time quantitative PCR. Expression of sig-naling molecules associated with innate immune response in virus-infected KMB17 and VK2 cells were ana-lyzed by real-time quantitative PCR. Results Both HSV1 and HSV2 could infect KMB17 and VK2 cells, and cause damage to cell morphology and inner structure after 12 hours. Both of the two viruses formed simi-lar plaque on the single layer of KMB17 and VK2 cells, although HSV2 proliferated slower than HSV1. There were differences in the expression of signaling molecules associated with innate immune response in-duced by the two viruses in KMB17 and VK2 cells. Conclusion Both HSV1 and HSV2 could infect and proliferate in epithelial cells ( KMB17 and VK2 cells) . Although there were slight differences in viral prolif-eration between them, significant differences in the expression of signaling molecules associated with innate immune response induced by the two viruses were observed.

Objective To provide a comprehensive reference index for different mouse models of herpes simplex virus 1 (HSV-1) infection by investigating the related clinical manifestations, pathological features and characteristics of viral distribution in tissues and organs of BALB/c mice infected with different HSV-1 strains by using different strategies.Methods Acute infection models were established by challenging BALB/c mice at age three or six weeks with HSV-1 17+ and McKrae strains via intranasal and corneal administrations.Correspondingly, chronic infection models were established with BALB/c mice through subcutaneous and foot pad injections.Results Although all experimental mice showed trichiasis and roachback, there were differences in weight and fatality rate among different groups.Results of the quantitative PCR detection indicated that the proliferation of HSV-1 in the nervous tissues (brain, spinal cord, trigeminal ganglion) varied among different groups.The pathological examination indicated that in the acute infection groups, significant pathological changes only occurred in the brain tissues, while in the chronic infection groups, pathological injuries only occurred in the trigeminal ganglia.Although a key index latency-associated transcript (LAT) was not detected in the trigeminal nerve tissues of mice in the chronic infection groups, co-culturing the tissues with Vero cells resulted in infectious lesions in the cells.Conclusion This study indicates that there are significant differences in weight and fatality rate among different BALB/c mouse models of HSV-1 infection.Varied replication dynamics of HSV-1 were observed in different tissues or organs of the BALB/c mice in different groups.Therefore, different indexes should be adopted to evaluate different HSV-1 infection models.