AIM:To observe the characteristics of estrous cycle of female ICR mice and ovariectomized mice that had reached sexual maturity stage .METHODS:ICR mice were randomly divided into 3 groups, normal group, sham operation group and ovariectomized group .After ovariectomy and sham operating , at the 8th day, the mice were taken vagi-nal smears twice daily (09:00 and 16:00) for 12 consecutive days.The feature of vulva in the mice and vaginal smear cy-tology, occurrence rate of regular estrous cycle , duration of estrous cycle and each stage duration of estrous cycle were re -corded and observed .RESULTS:The characteristics of estrous cycle in normal group and sham operation group were simi-lar.Nearly 85%of the ICR mice in sexual maturity stage showed regular estrous cycle .The majority manifested 6-day cy-cle (83%), and only a small number exhibited 5-day cycle (17%).Compared with 5-day group, the duration of oestrus was significantly increased in 6-day group (P<0.05).The number and kind of the cells in vaginal smear were a little dif-ferent between 5-day and 6-day groups.The ovariectomized mice which showed irregular estrous cycle (76%) were charac-terized by continuous diestrus .CONCLUSION:Female ICR mice have the strain characteristics of estrus cycle .

Objective:To analyze phylogenetic structure and molecular characteristics of H5N6 avian influenza virus (AIVs) isolated from live poultry market (LPM).Methods:Oropharyngeal and cloacal swabs from poultry, and environmental samples were collected from LPM in Urumqi in December 2018, AIVs were isolated and identified by inoculation of chicken embryo, hemagglutination test and RT-PCR, the viral whole genome was amplified with the universal primers of influenza A virus, and then sequenced, pairwise sequence alignments, phylogenetic and molecular characteristics analysis were performed by BLAST, Clustal W, MEGA-X and DNAStar software.Results:Six strains of H5N6 AIVs were isolated from poultry samples, the identity between the viral genes was high (99.4%-100.0%), so the isolates were the same source. BLAST analysis revealed that the viral NP sequence had the highest identity (99.7%) with H5N6 AIVs isolated from poultry in Suzhou, while the sequence of the remaining 7 viral genes had the highest identity (99.0%-100.0%) with H5N6 AIVs isolated from environment in Guangdong during 2017 to 2018. Phylogenetic analysis showed that the viral HA belonged to Clade 2.3.4.4C, and the viral HA, NA, PB1, PA, NP, and MP were all clustered together with H5N6 AIVs isolated from mink in Eastern China in 2018, while the PB2 and NS were clustered together with H5N6 AIVs isolated from environment in Guangdong from 2017 to 2018. The HA cleavage site contained multiple basic amino acid residues, which was highly pathogenic AIVs (HPAIVs). S137A and T160A mutations of HA could increase binding to human-type receptor SAα2, 6-Gal. Additionally, the viral multiple mutations, including 59-69 deletion in NA, the L89V, G309D, R477G, I495V, I504V, D391E, and A661E in PB2, as well as the P42S, D92E, and 80-84 deletion in NS1, could enhance the viral virulence and pathogenicity to mammals. Conclusions:The 6 strains of H5N6 HPAIVs isolated from LPM have relatively close genetic relationship with H5N6 AIVs isolated from mink in Eastern China and environment in Guangdong during 2017 to 2018, the viral multiple mutations could increase its pathogenicity to mammals, which could pose a potential risk to public health.

Objective:To study immunosuppression mediated by the porcine FcγRⅢ in porcine reproductive and respiratory syndrome virus ( PRRSV ) infection to pulmonary alveolar macrophages ( PAMS ).Methods: In this study pulmonary alveolar macrophages cells were treated with containing 200 TCID50 PRRSV,lipopolysaccharide (LPS) (100 ng/ml) and purified mouse anti-pig FcγRⅢIgG (550 μg/ml) separately,simultaneously,PAM cells treated with purified mouse anti-pig FcγRⅢIgG (550 μg/ml) was infected by 200 TCID50 PRRSV ,untreated PAM cells as the control group.Each group were post-cultured 12,24,36,48,60,72 h, the cells and the supernatant were collected.The dynamic variation of PRRSV RNA copies in inoculation group were detected by using real-time fluorescence quantitative PCR method.mRNA level of IFN-αand TNF-αin each group were detected by using relative fluorescence quantitative PCR.Results:The result showed that mRNA level of IFN-αwas improved during PRRSV infection to PAMS 12-24 h,and mRNA level of IFN-αwas inhibited during 36-72 h,then mRNA level of IFN-αrecovered normally; mRNA level of TNF-αwas increased slightly post-infection 12-72 h.IFN-αand TNF-αmRNA levels of PAM cells treated with LPS were both up-regu-lated,using the purified mouse anti-pig FcγRⅢ IgG to treat the PAM cells,selective activation of porcine FcγRⅢ in the PAM cells down-regulated significantly mRNA levels of IFN-αand TNF-α.PRRSV infection assay mediated by selective activation FcγRⅢof the PAM cells inhibited antiviral cytokine ( IFN-αand TNF-α) mRNA levels.Conclusion:The results show selective activation of FcγRⅢinhibited significantly mRNA levels of the antiviral cytokine IFN-αand TNF-αof host cells,and innate antiviral immune response to PRRSV infection.

Objective To investigate the value of stretched-exponential model of DWI in differential diagnosis of benign and malignant breast lesions.Methods Totally 58 patients with 63 breast lesions (33 benign,30 malignant lesions) were enrolled.All the patients underwent multiple b value DWI and dynamic contrast enhancement MRI (DCE-MRI) scans.The values of ADC,DDC and water molecular diffusion heterogeneity index (α) were calculated,and the time signal intensity curve (TIC) was obtained.All the parameters were compared between benign and malignant breast lesions.The diagnostic performance of different parameters was evaluated with ROC curve.Results ADC,DDC and α value of malignant lesions was (1.01±0.19)×10-3 mm2/s,(0.89±0.23)×10-3 mm2/s and 0.75±0.09,while of benign lesions was (1.41±0.27)× 10-3 mm2/s,(1.49±0.29)× 10-3mm2/s and 0.87±0.07,respectively.All 3 parameters in malignant lesions were lower than those in benign lesions (all P＜0.01).Taking 1.22 × 10-3 mm2/s as the optimal threshold,the area under the curve (AUC) of DDC was the largest as 0.958,and the corresponding diagnostic sensitivity and specificity was 96.67％ and 81.82％,respectively.AUC value was 0.976 by combining DDC with TIC,and the corresponding diagnostic sensitivity and specificity was 93.33％ and 93.94％,respectively.Conclusion The stretched-exponential model DWI can differentiate breast lesions,and diagnostic performance of combination of DDC and TIC is better than ADC or DCE.

Objective:To explore the potential categories of resourcefulness in primary caregivers of stroke patients and analyze the differences in the characteristics of different types of caregivers, so as to provide basis for clinical construction of effective intervention plans.Methods:The 308 primary caregivers of stroke patients who met the research standards in the Department of Neurology of General Hospital of Ningxia Medical University, People′s Hospital of Ningxia Hui Autonomous Region, the First People′s Hospital of Yinchuan from December 2020 to August 2021 were selected as the research objects by convenient sampling method.The general information questionnaire, Resourcefulness Scale and Social Support Revalued Scale were used for cross-sectional investigation.Results:The resourcefulness of the primary caregivers of stroke patients were divided into 2 potential categories: low resourcefulness group (62.7%, 193/308) and high resourcefulness group (37.3%, 115/308). Uni-variate analysis showed that there were differences between potential categories: age and education level of stroke patients, whether there was long-term medication history, number of other chronic diseases, had there been any complications due to illness, age, education level and occupation of caregivers, average daily care duration, family relations, and social support ( χ2 values were -3.34 - 23.62, all P<0.05). Logistic regression analysis showed that social support and education level of caregivers were the influencing factors of caregiver resourcefulness ( P<0.05). Conclusions:The resourcefulness in the primary caregivers of stroke patients has certain characteristics. Clinically, targeted intervention measures should be formulated according to the different types of caregiver resourcefulness characteristics of caregivers and their influencing factors to improve the caregiver resourcefulness of stroke patients.

Objective To explore the research context,hotspots and development direction of eye acupuncture therapy through analysis on relevant literature about eye acupuncture therapy based on visualization analysis on knowledge map.Methods The literature related to eye acupuncture therapy was retrieved from CNKI,Wanfang Data,VIP and CBM from the establishment of the databases to 2022.NoteExpress 3.7.0.9296 was used to screen the titles.WPS 11.1.0.13703 was used to analyze the time of publications and the source journals,and CiteSpace 6.1.R6 was used to analyze the authors,institutions and keywords included in the literature.Results A total of 964 articles were included after screening.The total number of publications showed an increasing trend of fluctuation.187 journals were involved,and the journals with the highest number of articles were Liaoning Journal of Traditional Chinese Medicine,Chinese Acupuncture and Moxibustion,Practical Journal of Internal Medicine of Traditional Chinese Medicine;805 high-yield authors such as Wang Pengqin,Wang Jian and Wang Zhe were included.369 institutions including Liaoning University of Traditional Chinese Medicine,Guangzhou University of Traditional Chinese Medicine and Tianjin University of Traditional Chinese Medicine were included.A total of 637 keywords such as stroke,cerebral ischemia-reperfusion injury,and clinical research were included.Conclusion The research of eye acupuncture therapy has gradually developed from theoretical interpretation,case report,curative effect observation and experience of famous doctors to basic research and higher-level clinical research of dominant diseases.Post-stroke mental disorders and brain diseases excluding cerebrovascular diseases are the research trends in this field.

OBJECTIVE: To discover the relationship of transcriptional levels and promoter methylation status of CHFR gene in human nasopharyngeal carcinoma,to discuss the significance and epigenetic mechanism of CHFR inactivation in NPC, and to evaluate the feasibility of detecting methylated CHFR in nasopharyngeal swab as a means for diagnosis of NPC. METHOD: Transcriptional levels of CHFR was evaluated by RT-PCR. Methylation specific PCR was used to detect the methylation status of CHFR in NPC cells, normal nasopharyngeal epithelia, primary tumors and their paired nasopharyngeal swabs. Detailed methylation status was confirmed by bisulfite sequencing. NPC cells were treated by the methyltransferase inhibitor 5-aza-dC and the reactivation of CHFR was evaluated by RT-PCR. RESULT: CHFR transcription was inactivated in NPC. The methylation frequency in NPC primary tumors and their paired swabs were 65.5% and 63.8%, respectively, with a 86.2% concordance. Bisulfite sequencing revealed a dense methylation in NPC cells and primary tumors, but all the normal nasopharyngeal epithelia were unmethylated. CHFR expression were restored after 5-aza-dC treatment. CONCLUSION CHFR is epigenetically inactivated by promoter methylation in NPC. Detecting methylated CHFR can be served as a useful non-invasive means for diagnosis of NPC.
Aged ; Carcinoma ; Cell Cycle Proteins ; genetics ; DNA Methylation ; Epigenesis, Genetic ; Female ; Gene Silencing ; Humans ; Male ; Middle Aged ; Nasopharyngeal Carcinoma ; Nasopharyngeal Neoplasms ; genetics ; pathology ; Neoplasm Proteins ; genetics ; Neoplasm Staging ; Poly-ADP-Ribose Binding Proteins ; Promoter Regions, Genetic ; Ubiquitin-Protein Ligases

Objective:To explore the relationship between psychological distress and intimacy in stroke patients and the mediating effect of different dyadic coping style, to provide theoretical guidance for improving the intimate relationship of stroke patients.Methods:A convenient sampling method was used to select 203 patients with stroke who visited the General Hospital of Ningxia Medical University, the People's Hospital of Ningxia Hui Autonomous Region and the Department of Neurology of Yinchuan First People's Hospital from December 2020 to July 2021 as the survey objects. The general information questionnaire, Kessler Psychological Distress Scale, Quality of Relationship Index and Dyadic Coping Inventory were used to investigate patients.Results:The scores of psychological distress, intimate relationship, positive dyadic coping and negative dyadic coping were 19.33 ± 7.46, 32.75 ± 6.79, 86.25 ± 13.22, and 16.46 ± 4.08, respectively. Correlation analysis showed that psychological distress was negatively correlated with intimate relationship and positive dyadic coping ( r=-0.195, -0.204, both P<0.01), psychological distress was positively correlated with negative dyadic coping ( r=0.229, P<0.01). The mediating effect of positive dyadic coping and negative dyadic coping on psychological distress and intimate relationship was 35.87% and 53.26%, respectively. Conclusions:The psychological distress indirectly affects the intimate relationship of stroke patients through positive dyadic coping and negative dyadic coping. Health care workers can guide patients to actively cope with the disease and encourage couples to strengthen communication, so as to enhance the intimacy of couples and promote their physical and mental health development.

Objective:To construct a conceptual framework of burnout among primary caregivers of stroke patients based on grounded theory.Methods:According to the characteristics of age, gender, relationship with patients, length of care and other characteristics of purpose sampling method, from December 2021 to March 2022, 31 primary caregivers of stroke patients were selected for semi-structured interviews in two second-class hospitals, a first-class hospitals and a community health service station in Wuzhong city and Yinchuan city. The data were analyzed in procedural grounded theory method.Results:A total of 29 primary categories and 9 secondary categories were formed. The conceptual framework of burnout among primary caregivers of stroke patients was finally established, which consisted of four main categories (role burnout, physical burnout, psychological burnout and social burnout) and a core category (burnout of primary caregivers of stroke patients).Conclusions:The conceptual framework of burnout among primary caregivers of stroke patients constructed in this study elaborates the connotation of burnout of primary caregivers of stroke patients from four aspects, which can provide a basis for the development of assessment tools and the formulation of intervention measures in the future.

Objective To construct c-Met CAR-T cells secreting PD-1 antibodies to reduce immune inhibitory effect of tumor cells and enhance the efficacy of CAR-T cell therapy against pancreatic cancer.Methods Kaplan-Meier Plotter,GEPIA,and Timer 2.0 bioinformatics databases were used to analyze c-Met expression in pancreatic cancer and its correlation with survival and immune infiltration status.In clinical samples of pancreatic cancer and pancreatic cancer Aspc-1 cells,c-Met and PD-L1 expressions were detected using immunohistochemistry or flow cytometry.Using gene editing technology,PD-1 secretory antibodies and HIS tags were linked to second-generation c-Met CAR molecules to construct PD-1/c-Met CAR plasmids,which were then packaged into lentiviruses for infection of activated T cells.The positive rate and cell subset distribution of CAR-T cells were analyzed with flow cytometry,and secretory PD-1 antibodies in cell supernatants were detected using Western blotting.The target cell killing efficiency and proliferative activity of the modified CAR-T cells were evaluated after activation,and cytokine secretion was analyzed using ELISA.Results The expression of c-Met was significantly higher in pancreatic cancer than in normal tissues,and its expression level was negatively correlated with the patients'survival and positively correlated with immune cell infiltration.The clinical samples of pancreatic cancer tissues expressed significantly higher levels of c-Met and PD-L1 than the adjacent tissues,and 90.7%and 57.7%of Aspc-1 cells were positive for c-Met and PD-L1,respectively.The constructed PD-1/c-Met CAR-T cells were capable of secreting PD-1 antibodies and showed a significantly higher killing efficiency against tumor cells than c-Met CAR-T cells at an effector-to-target ratio of 20:1,with also a higher proliferative activity after target cell stimulation and higher levels of IL-2 and TNF-α secretin.Conclusion PD-1/c-Met CAR-T cells have higher killing efficiency against pancreatic cancer cells with also higher proliferative activity than c-Met CAR-T cells.