To investigate the therapeutic effect and related mechanisms of hordenine on ovalbumin (OVA)-induced allergic rhinitis (AR) in rats, HE and AB-PAS staining were used to detect the improvement of pathological damage to the nasal mucosa induced by hordenine. ELISA was employed to detect the effect of hordenine on OVA-sIgE in serum and IL-4 in the nasal mucosa supernatant of rats. IHC and Western blot experiments were undertaken to examine the effect of hordenine on Th1/Th2 cell balance. Bioinformatics analysis was performed to predict pathways, which were verified by in vivo and in vitro experiments. The experimental results showed that hordenine could alleviate the behavioral manifestations of OVA-induced AR rats, alleviate nasal mucosal pathological damage caused by AR, and reduce the secretion of OVA-sIgE and IL-4. In addition, hordenine could regulate the Th1/Th2 balance. Bioinformatics analysis results showed that the potential pathway of action of hordenine on AR was the phosphoinositide 3 kinase (PI3K)/protein kinase B (Akt) signaling pathway. The in vivo experimental results showed that the expression of PI3K and p-Akt proteins in the nasal mucosa of the model group rats was significantly increased (P < 0.01), and that the protein expression level was significantly decreased after the administration of hordenine, which was also confirmed by an in vitro experiment. This study suggests that hordenine may regulate Th1/Th2 cell balance through the PI3K/Akt signaling pathway, thereby exerting an alleviating effect on OVA-induced AR.

Objective To analyze the status of persistent human papillomavirus (HPV) infection and the distribution of viral subtypes in the Zhengzhou region. Methods Clinical data of 55239 patients who underwent HPV-DNA genotyping tests from 2018 to 2024 were collected. On the basis of HPV follow-up results, patients were divided into three groups: 849 cases of infection with the same HPV type, 255 cases of persistent infection with different HPV types, and 857 cases of transient HPV infection. Results Among the 9232 initially-screened patients who were positive with HPV, the high-risk HPV (HR-HPV) positive rate was 84.6% (7813/9232), and predominant subtypes were HPV-16 (20.8%), HPV-52 (17.2%), and HPV-53 (9.6%). The low-risk HPV (LR-HPV) positive rate was 15.4% (1419/9232), mainly HPV-81 (28.1%) and HPV-42 (27.0%). Among the 1961 follow-up cases, the rates of persistent and non-persistent infections with the same HPV type were 35.7% (701/1961) and 7.5% (148/1961), respectively. The rates of persistent and non-persistent infections with different HPV types were 10.9% (214/1961) and 2.1% (41/1961), respectively. Meanwhile, 43.7% (857/1961) tested negative upon follow-up. Among the 701 cases of persistent infection with the same HPV type, HR-HPV accounted for 85.7% (601/701), and LR-HPV accounted for 14.3% (100/701). Among the 214 cases of persistent infection with different HPV types, 89.7% (192/214) were high-risk transitions, and 10.3% (22/214) were low-risk transitions. Regardless of HPV types, HR-HPV was more likely to cause persistent infection than LR-HPV. In addition, over 60% of HR-HPV persistent infections resolved within 18 months, 30% developed into persistent infections, and only 15% of patients had persistent infections that last more than 24 months. Conclusion Persistent HPV infections are predominantly high-risk types. Most patients clear the infection within 18 months, approximately 30% develop persistent infections, and about 15% of patients have persistent infections that last more than 24 months. However, the HR-HPV persistent infection rates across different age groups slightly differ.

Objective To investigate the cell membrane-penetrating capacity of human cell-penetrating peptide hPP10 carrying human antioxidant protein Cu-Zn superoxide dismutase(Cu,Zn-SOD)and assess the antioxidant and anti-inflammatory activity of these fusion proteins.Methods The fusion protein hPP10-Cu,Zn-SOD was obtained by genetic engineering and identified by Western blotting.The membrane-penetrating ability of the fusion protein was evaluated by immunofluorescence assay,fluorescence colocalization assay and Western blotting,its SOD enzyme activity was detected using a commercial kit,and its effect on cell viability was assessed with MTT assay.In a HEK293 cell model of H2O2-induced oxidative stress,the effect of hPP10-Cu,Zn-SOD on cell apoptosis was analyzed with flow cytometry and RT-qPCR,and its antioxidant effect was assessed using reactive oxygen species(ROS)assay;its anti-inflammatory effect was evaluated in mouse model of TPA-induced ear inflammation by detecting expression of the inflammatory factors using RT-qPCR,Western blotting and immunohistochemistry.Results The fusion protein hPP10-Cu,Zn-SOD was successfully obtained.Immunofluorescence assay confirmed obvious membrane penetration of this fusion protein in HEK293 cells,localized both in the cell membrane and the cell nuclei after cell entry.hPP10-Cu,Zn-SOD at the concentration of 5 μmol/L exhibited strong antioxidant activity with minimal impact on cell viability at the concentration up to 10 μmol/L.The fusion protein obviously inhibited apoptosis and decreased intracellular ROS level in the oxidative stress cell model and significantly reduced mRNA and protein expression of the inflammatory factors in the mouse model of ear inflammation.Conclusion The fusion protein hPP10-Cu,Zn-SOD capable of penetrating the cell membrane possesses strong antioxidant and anti-inflammatory activities with only minimal cytotoxicity,demonstrating the value of hPP10 as an efficient drug delivery vector and the potential of hPP10-Cu,Zn-SOD in the development of skincare products

Objective To investigate the cell membrane-penetrating capacity of human cell-penetrating peptide hPP10 carrying human antioxidant protein Cu-Zn superoxide dismutase(Cu,Zn-SOD)and assess the antioxidant and anti-inflammatory activity of these fusion proteins.Methods The fusion protein hPP10-Cu,Zn-SOD was obtained by genetic engineering and identified by Western blotting.The membrane-penetrating ability of the fusion protein was evaluated by immunofluorescence assay,fluorescence colocalization assay and Western blotting,its SOD enzyme activity was detected using a commercial kit,and its effect on cell viability was assessed with MTT assay.In a HEK293 cell model of H2O2-induced oxidative stress,the effect of hPP10-Cu,Zn-SOD on cell apoptosis was analyzed with flow cytometry and RT-qPCR,and its antioxidant effect was assessed using reactive oxygen species(ROS)assay;its anti-inflammatory effect was evaluated in mouse model of TPA-induced ear inflammation by detecting expression of the inflammatory factors using RT-qPCR,Western blotting and immunohistochemistry.Results The fusion protein hPP10-Cu,Zn-SOD was successfully obtained.Immunofluorescence assay confirmed obvious membrane penetration of this fusion protein in HEK293 cells,localized both in the cell membrane and the cell nuclei after cell entry.hPP10-Cu,Zn-SOD at the concentration of 5 μmol/L exhibited strong antioxidant activity with minimal impact on cell viability at the concentration up to 10 μmol/L.The fusion protein obviously inhibited apoptosis and decreased intracellular ROS level in the oxidative stress cell model and significantly reduced mRNA and protein expression of the inflammatory factors in the mouse model of ear inflammation.Conclusion The fusion protein hPP10-Cu,Zn-SOD capable of penetrating the cell membrane possesses strong antioxidant and anti-inflammatory activities with only minimal cytotoxicity,demonstrating the value of hPP10 as an efficient drug delivery vector and the potential of hPP10-Cu,Zn-SOD in the development of skincare products

Objective To explore the clinical efficacy of repeated transcranial magnetic stimulation(rTMS)and acupuncture therapy in the treatment of chronic insomnia disorder(CID)patients.Methods A total of 80 patients with CID,who were treated at Nanchang First Hospital from January 2022 to December 2023,were selected for the study.The patients were randomly divided into control group and treatment group,with 40 cases in each group.The control group patients were treated with dexmedetomidine,while the treatment group patients received rTMS and acupuncture therapy in addition to control group.The treatment course was 4 weeks,and the sleep quality,sleep related indicators,and psychological condition improvement of both groups of patients were observed before and after treatment.Results After treatment,the Pittsburgh sleep quality index scores of both groups of patients decreased(P＜0.05);The sleep latency and number of awakenings were lower than before treatment(P＜0.05),and the total sleep time,sleep efficiency,and proportion of rapid eye movement sleep were higher than before treatment,treatment group showed more significant improvement than control group(P＜0.05).After treatment,the Hamilton anxiety and depression scale scores of both groups of patients decreased compared to before treatment,but there was no statistically significant difference in control group before and after treatment(P＞0.05).However,there was a statistically significant difference in treatment group before and after treatment(P＜0.05).Conclusion The combination of rTMS and acupuncture treatment can significantly improve the sleep quality of CID patients,while also reducing the accompanying symptoms of anxiety and depression.

Objective:To establish the "1+3" management module of high-value consumables in Operation Room and verify its application, so as to provide new ideas for cost management of consumables in Operation Room.Methods:The Operating Room team of Shenzhen Hospital of Southern Medical University designed a "1+3" management module in 2022, where "1" referred to the management process of high-value consumables in Operation Room, and "3" referred to the precise management of consumables in Operation Room warehouse, the management of closed-loop use of Operation Room consumables and adverse event management of consumables. Surgeries using high-value consumables in the Thoracic Surgery Department, Gastrointestinal Surgery Department, and Urology Department of the hospital were selected as the research objects. The surgeries using conventional consumables from January to June 2022 were set as the control group, and the surgeries implementing the "1+3" management module from July to December 2022 were set as the observation group. The number of consumables received by the itinerant nurses before the operation and the number of high-value consumables returned after the operation were compared between the two groups. And the number of missed and error charges for high-value consumables in the two groups were counted and compared.Results:The number of consumables received before operation in the control group was higher than that in the observation group, and the difference was statistically significant ( P<0.05). The number of high-value consumables returned in the observation group was less than that in the control group, and the difference was statistically significant ( P<0.01). The proportion of missed charges for consumables in the observation group was lower than that in the control group, and the difference was statistically significant ( P<0.01), but there was no statistically significant difference in the proportion of incorrect charges between the two groups ( P>0.05) . Conclusions:The "1+3" management module for high-value consumables in Operation Room makes the process of receiving, returning, and charging high-value consumables clear, with traceable data, achieving refined management of high-value consumables in Operation Room, reducing the number of high-value consumables returned to the warehouse and reducing the proportion of missed consumables, which is conducive to effective cost control in Operation Room.

Animal experiments play an important role in the process of biomedical research, and is a necessary way to transform basic medicine into clinical medicine. The standardization of animal experimental studies and reports determines the reliability and reproducibility of research results, and is also the key to transforming the results of animal experiments into clinical trials. In view of how to design and implement animal experiments, write animal experiment reports, and publish relevant academic papers in a more standardized way, LACM (Laboratory Animal and Comparative Medicine) has launched a new column of comparative medical research and reporting standards from 2023, focusing on the introduction and interpretation of international general norms related to laboratory animal and comparative medicine, such as ARRIVE 2.0 guidelines (Animal Research: Reporting of In Vivo Experiments). This article focuses on the development and application, basic content and priority of ARRIVE 2.0, as well as the scheme of implementing ARRIVE 2.0 guidelines in international biomedical journals, and explains the current situation and future plans of LACM following ARRIVE 2.0 guidelines. The research and report of animal experimental medicine following the ARRIVE 2.0 guidelines and other international norms is one of the important driving forces to promote the high-quality development of experimental animal science and biomedicine in China, and also a powerful means to implement the 3R principle and improve the welfare of laboratory animals. Through this article, we hope the majority of scientific researchers and editors will attach great importance and actively implement these international standards.

Objective:To investigate the screening values of immunocytochemical P16/Ki-67 double staining, P16 INK4α single staining and high-risk human papillomavirus (HR-HPV) testing for high-grade cervical lesions. Methods:The clinical data of 622 patients who underwent cervical thin-layer liquid-based cytology (TCT) and HR-HPV testing in General Hospital of Taiyuan Iron and Steel (Group) Co., Ltd. from March 2019 to July 2021 were retrospectively analyzed. The remaining cytological specimens were detected by P16/Ki-67 double staining and P16 INK4α single staining. Among them, 334 patients with TCT results suggesting atypical squamous cell of undetermined significance (ASCUS) and above and HPV-positive underwent colposcopy pathological biopsy. Using pathological results as reference, the positive predictive value, sensitivity, specificity and accuracy of P16/Ki-67 double staining, P16 INK4α single staining and HR-HPV testing for screening of high-grade squamous intraepithelial neoplasia (HSIL) and cervical cancer were compared. Results:Taking the results of histopathology as references, combined with the results of TCT, 31 of 622 patients were HSIL, of which 22 (71.0%) were positive for P16/Ki-67 double staining, 23 (74.2%) were positive for P16 INK4α single staining, and 25 (80.6%) were positive for HR-HPV testing; 4 cases were cervical cancer, and the positive rates of the three detection methods were all 100.0% (4/4). Among 622 patients, the positive rates of P16/Ki-67 double staining, P16 INK4α single staining and HR-HPV testing for screening of HSIL and cervical cancer were 13.99% (87/622), 25.40% (158/622) and 21.38% (133/622); the positive predictive values were 29.89%, 17.09% and 21.08%; the accuracies were 91.19%, 78.94% and 83.28%; the specificities were 89.77%, 77.98% and 82.46%; the sensitivities were 74.29%, 77.14% and 82.86%. The positive rate, positive predictive value, specificity and accuracy of P16/Ki-67 double staining were higher than those of P16 INK4α single staining and HR-HPV testing, and the differences were statistically significant ( z values were -5.062 and -3.418, 2.328 and 2.450, 5.436 and 3.570, 6.043 and 4.161, all P < 0.05); the sensitivity of HR-HPV testing was higher than that of P16/Ki-67 double staining and P16 INK4α single staining, but the differences were not statistically significant ( z values were -0.890 and 1.017, both P > 0.05). Conclusions:HR-HPV testing is more suitable for primary cervical lesion screening; P16/Ki-67 double staining can be used as a potential combined cell screening tool or an effective triage tool; P16 INK4α single staining has certain limitations.

To knock out β-lactoglobulin (BLG) gene and insert human lactoferrin (hLF) coding sequence at BLG locus of goat, the transcription activator-like effector nucleases (TALEN) mediated recombination was used to edit the BLG gene of goat fetal fibroblast, then as donor cells for somatic cell nuclear transfer. We designed a pair of specific plasmid TALEN-3-L/R for goat BLG exon III recognition sites, and BLC14-TK vector containing a negative selection gene HSV-TK, was used for the knock in of hLF gene. TALENs plasmids were transfected into the goat fetal fibroblast cells, and the cells were screened three days by 2 μg/mL puromycin. DNA cleavage activities of cells were verified by PCR amplification and DNA production sequencing. Then, targeting vector BLC14-TK and plasmids TALEN-3-L/R were co-transfected into goat fetal fibroblasts, both 700 μg/mL G418 and 2 μg/mL GCV were simultaneously used to screen G418-resistant cells. Detections of integration and recombination were implemented to obtain cells with hLF gene site-specific integration. We chose targeting cells as donor cells for somatic cell nuclear transfer. The mutagenicity of TALEN-3-L/R was between 25% and 30%. A total of 335 reconstructed embryos with 6 BLG-/hLF+ targeting cell lines were transferred into 16 recipient goats. There were 9 pregnancies confirmed by ultrasound on day 30 to 35 (pregnancy rate of 39.1%), and one of 50-day-old fetus with BLG-/hLF+ was achieved. These results provide the basis for hLF gene knock-in at BLG locus of goat and cultivating transgenic goat of low allergens and rich hLF in the milk.
Animals ; Animals, Genetically Modified ; genetics ; Female ; Fibroblasts ; Gene Knock-In Techniques ; Gene Knockout Techniques ; Goats ; genetics ; Humans ; Lactoferrin ; genetics ; Lactoglobulins ; genetics ; Milk ; chemistry ; Nuclear Transfer Techniques ; Plasmids ; Pregnancy ; Transfection

Objective To analyze the clinical characteristics of patients with L isteria septicemia for better clinical diagnosis and management of the disease .Methods A retrospective study was carried out in 13 patients with confirmed diagnosis of Listeria septicemia from July 2009 to November 2013 in West China Hospital of Sichuan University .The clinical features ,laboratory tests ,treatments and clinical outcomes were reviewed and analyzed . Results The vast majority of the 13 patients were immunocompromised or with critical organ dysfunction . The in vitro antimicrobial susceptibility testing indicated that penicillin ,ampicillin and levofloxacin were the most active agents against Listeria ,followed by imipenem ,erythromycin , ciprofloxacin and tetracycline .Only 33 .3% of the 13 Listeria isolates were sensitive to oxacillin .Eight patients were cured ,2 improved ,2 died after therapy .The remaining one patient gave up therapy .Conclusions The incidence of Listeria septicemia was associated with advanced age and presence of underlying diseases .Early etiology diagnosis and appropriate antibacterial therapy can improve the outcome of such patients .Actively treating underlying diseases helps reduce the mortality rate .