ObjectiveTo establish a highly specific and sensitive triplex TaqMan quantitative PCR method for the detection of mouse hepatitis virus (MHV), mouse parvovirus (MPV), and reovirus type 3 (Reo-3) in laboratory mice. MethodsRecombinant plasmid standards were constructed using conserved genomic regions of the three target viruses. Specific primers and probes were designed, and a triplex TaqMan quantitative PCR (qPCR) system was optimized through preliminary experiments. Sensitivity was evaluated using 10-fold serial dilutions (10¹-10⁷ copies/μL). Intra- and inter-assay repeatability were assessed via reproducibility experiments. ResultsA strong linear correlation (R ²>0.99) was observed between copy number and Ct value at template concentrations of 10²-10⁷ copies/μL. Intra-assay and inter-assay coefficients of variation (CV) were both below 5%. No cross-reactivity was observed with mouse cytomegalovirus (MCMV), vesicular stomatitis virus (VSV), Sendai virus (SeV), or pneumonia virus of mice (PVM). ConclusionThe triplex TaqMan quantitative PCR method established in this study has high sensitivity, good repeatability, and strong specificity. It enables the rapid, specific, and sensitive detection of MHV, MPV, and Reo-3, allows the simultaneous detection of multiple pathogens in a single tube, and can be applied to the detection of clinical samples from laboratory animals and to epidemiological investigations, thereby providing effective scientific and technological support for the prevention and control of viral infection and the interruption of transmission.

Purpose: This study aimed to evaluate the safety and efficacy of gemcitabine and cisplatin (GP) regimen in combination with immune checkpoint inhibitor sintilimab as neoadjuvant therapy for muscle-invasive bladder cancer (MIBC) patients and the feasibility of the following selective bladder sparing surgery. Materials and Methods: Patients with histopathologically confirmed urothelial carcinoma without distant metastases (T2-4a, N ≤ 1, M0, American Joint Committee of Cancer 8th) and with adequate organ function will be enrolled. The therapeutic regimen was sintilimab 200 mg once on day 8, gemcitabine 1,000 mg/m2 and cisplatin 35 mg/m2 once on days 1 and 8, every 21 days for four cycles. The primary endpoint was pathologic complete response (pCR, pT0N0) rate. The secondary end points were ypT < 2 rate, R0 resection rate, event-free survival, and safety. Results: From May 4, 2020, to May 20, 2023, 55 patients were enrolled. Forty-six patients were evaluated for efficacy. Among the 42 patients who underwent surgery, 16 patients (38.0%) achieved pCR. Thirty-three patients (78.6%) achieved pT < 2. With a median follow-up of 15.7 months, the 1-year event-free survival was 91.3%. Notwithstanding the poor pathological baseline characteristic of a high T3-T4a proportion (39.1%), a promising bladder preservation (including 22 patients transurethral resection of bladder tumor, 5 patients partial cystectomy, and 4 surveillances) rate was achieved (67.4%). The most common grade ≥ 3 treatment-related adverse events was neutropenia (n=15, 27.3%), which was related to chemotherapy. There were no grade 3 immune-related adverse events. Conclusion Neoadjuvant GP plus sintilimab is a promising regimen for MIBC patients, with relatively high pT < 2 rate and triggering the emerging roles for the multi-disciplinary team decision-making for bladder sparing surgery.

Objective To develop a Discharge Readiness Scale for Patients with PICC and to test its reliability and validity.Methods Based on the Meleis transition theory,the initial version of the scale was formed through literature review,qualitative interview,expert enquiry and pre-investigation.Patients with PICC in 2 tertiary hospital in Hangzhou from December 2023 to March 2024 were selected to investigate the reliability and validity of the scale by convenience sampling.Results 352 questionnaires were distributed,and 340 valid questionnaires were recovered,and the validity recovery rate was 96.59％.The exploratory factor analysis extracted 4 common factors with the cumulative variance contribution rate of 80.568％.The confirmatory factor analysis showed that all evaluation indicators of the scale were within ideal standards.The item-level content validity index ranged from 0.79 to 1.00,and the scale-level content validity index was 0.96.The total Cronbach's α coefficient of the scale was 0.963,and the split-half reliability coefficient was 0.889.The Chinese version of the Discharge Readiness Scale was used as a calibration tool,and the correlation coefficients between the total score of the scale and the scores of each dimension and the calibration tool were 0.788 and 0.567～0.878,respectively(all P＜0.05).The Discharge Readiness Scale for Patients with PICC included 26 items in 4 dimensions including physiological and psychological status,knowledge of catheter maintenance,catheter maintenance capability and social support.Conclusion The Discharge Readiness Scale for Patients with PICC has good reliability and validity,and can be used to evaluate the level of discharge readiness among patients with PICC.

BACKGROUND:The progression of chronic oral diseases is closely related to the continuous inflammatory response.Anti-inflammatory peptides are expected to become a substitute for traditional anti-inflammatory drugs due to their rich sources,easy absorption by the body and low side effects.OBJECTIVE:To review the types,anti-inflammatory mechanisms,and their application in oral related diseases.METHODS:CNKI,PubMed,and Web of Science were retrieved,with"polypeptide,anti-inflammatory,immunomodulation,oral inflammatory diseases"as Chinese and English search terms.111 articles related to the classification of anti-inflammatory peptides,anti-inflammatory mechanisms,and application of oral inflammatory diseases were selected for review.RESULTS AND CONCLUSION:(1)Anti-inflammatory peptides are abundant in nature,which can be extracted from plants,animals,and microorganisms.In addition to naturally occurring peptides and protein hydrolysates,peptides synthesized by chemical modification,computer simulation design,and genetic recombination technology can also exert anti-inflammatory effects.The composition,position,and properties of amino acids affect their anti-inflammatory activity.(2)Because the anti-inflammatory mechanism of anti-inflammatory peptides is still unclear,the activity verification is mostly cell experiments,and there is a lack of animal models,clinical trials and other further studies.(3)In the treatment of oral inflammatory diseases(including periodontitis,oral mucositis,caries,pulpitis,suppurative osteomyelitis of the jaw,and peri-implantitis),anti-inflammatory peptides can inhibit the release of inflammatory factors such as interleukin 6,interleukin 1β,and tumor necrosis factor α in oral tissues,regulate inflammatory responses,improve the chronic inflammatory environment,reduce tissue destruction and structural loss,and promote bone tissue regeneration,providing new ideas for the treatment of oral inflammatory diseases.

Objectives:To evaluate the biomechanics of a new self-developed bidirectional self-locking fusion device in vitro.Methods:12 fresh L3/4 segment specimens of domestic pig spine were randomly divided into 3 groups for modeling:normal group(intact state),traditional fusion device group(traditional fusion device and unilateral screw fixation)and new fusion device group(bidirectional self-locking fusion device implanted).On the basis of MTS multi-freedom(spinal motion)simulation test system and 500N preload,8N·m load was applied to experimental specimens in each direction successively,and the stability test was carried out according to the order of forward bending,backward extension,left bending,right bending,and left and right rotation.The ranges of motion(ROMs)were compared between groups under different working conditions.The axial fatigue compression test was carried out with TT-JQ-01-106 fatigue testing machine to evaluate the stability and reliability of the self-developed fusion device.Results:Compared with the normal group,the ROM of the new fusion device group was significantly decreased under the conditions of flexion(1.67°±0.65° vs 3.46°±0.16°),extension(1.43°±1.01° vs 1.89°±0.29°),left bending(2.07°±1.32° vs 3.08°±0.09°),right bending(1.38°±0.72° vs 4.37°±0.20°),and right rotation(2.17°±1.51° vs 5.10°±0.37°)(all P＜0.05);Under the left rotation condition,the ROM value(3.10°±1.57°)of the new device group was significantly higher than that of the normal group(2.36°±0.62°)(P＜0.05).Compared with the conventional fusion device group,the ROM of the new fusion device group was significantly increased under flexion(1.67°±0.65° vs 1.22°±0.76°)and left rotation(3.10°±1.57° vs 1.57°±0.91°)conditions(P＜0.05);However,there were no significant differences in ROMs between the two groups under extension(1.43°±1.01° vs 1.06°±0.50°),left bending(2.07°±1.32° vs 1.87°±1.41°),right bending(1.38°±0.72° vs 1.45°±0.69°),and right rotation(2.17°±1.51° vs 1.69°±1.07°)conditions(P＞0.05).The ROM values of the normal group under flexion,extension,left bending,right bending,and right rotation conditions were significantly higher than those of the other two groups(P＜0.05).Fatigue test:after 5 million load-displacement cycles,visual inspection revealed no deformation or cracks in the fusion device,and the axial compression fatigue test was passed.Conclusions:The bidirectional self-locking fusion device can obviously improve the stability of the fusion segment,which is equivalent to the stability of using the posterior single-sided nail rod system,meanwhile,it has good fatigue resistance,therefore its mechanical properties can meet the needs of clinical applications.

Objective To develop a Discharge Readiness Scale for Patients with PICC and to test its reliability and validity.Methods Based on the Meleis transition theory,the initial version of the scale was formed through literature review,qualitative interview,expert enquiry and pre-investigation.Patients with PICC in 2 tertiary hospital in Hangzhou from December 2023 to March 2024 were selected to investigate the reliability and validity of the scale by convenience sampling.Results 352 questionnaires were distributed,and 340 valid questionnaires were recovered,and the validity recovery rate was 96.59％.The exploratory factor analysis extracted 4 common factors with the cumulative variance contribution rate of 80.568％.The confirmatory factor analysis showed that all evaluation indicators of the scale were within ideal standards.The item-level content validity index ranged from 0.79 to 1.00,and the scale-level content validity index was 0.96.The total Cronbach's α coefficient of the scale was 0.963,and the split-half reliability coefficient was 0.889.The Chinese version of the Discharge Readiness Scale was used as a calibration tool,and the correlation coefficients between the total score of the scale and the scores of each dimension and the calibration tool were 0.788 and 0.567～0.878,respectively(all P＜0.05).The Discharge Readiness Scale for Patients with PICC included 26 items in 4 dimensions including physiological and psychological status,knowledge of catheter maintenance,catheter maintenance capability and social support.Conclusion The Discharge Readiness Scale for Patients with PICC has good reliability and validity,and can be used to evaluate the level of discharge readiness among patients with PICC.

Objective:To construct a communication skills assessment scale for newly employed nurses, verify the reliability and validity of the scale in a simulated setting, and develop and construct an effective and structured assessment tool for the communication skills of newly employed nurses.Methods:The Chinese version of the Liverpool Communication Skills Assessment Scale was modified and two rounds of expert consultation were conducted to construct the communication skills assessment scale for newly employed nurses. A total of 194 newly employed nurses at a tertiary hospital between 2024 and 2025 were selected using convenience sampling. Data were collected using a teacher-based evaluation method during simulated communication scenarios. The performance of the nurses was scored, and the reliability and validity of the scale were analyzed.Results:The final version of the scale consisted of 4 dimensions and 11 items, and can be used in both workplace-based and simulation-based evaluations. The expert authority coefficients of both rounds were greater than 0.70. The Kendall's W coordination coefficients for the two rounds of consultation were 0.278 and 0.309 for workplace-based evaluations and 0.256 and 0.295 for simulation-based evaluations. The coefficients of variation for the 11 items in both application scenarios were <0.250. The total Cronbach's alpha coefficient of the scale was 0.805 and the total split-half reliability coefficient was 0.814. In the two application scenarios, the item-level content validity index ranged from 0.769 to 1.000 (all >0.750). The scale-level content validity index was 0.916 and 0.909 (>0.900), respectively, in the workplace-based and simulation-based evaluations. The exploratory factor analysis extracted a total of four common factors, with a cumulative variance contribution of 69.09%, and all item loadings on their corresponding factors exceeded 0.500. Conclusions:The communication skills assessment scale for newly employed nurses has moderate and validated content and number of items. The scale demonstrates high reliability and validity in simulation-based evaluations, and can be used as an effective tool for assessing the communication skills of newly employed nurses.

This study innovatively incorporates on-site teaching with the International Museum of Stomatology into the curriculum to establish an immersive and intuitive teaching mode,promoting education from both theoretical and practical dimensions.The teaching effect is comprehensively evaluated to explore the pathway to optimization.Multi-dimensional questionnaires are designed to collect feedback data from students on teaching satisfaction,knowledge mastery,professional identity,and humanistic literacy perception,fol-lowed by in-depth quantitative and qualitative analyses.The results demonstrate that this teaching mode significantly enhances literacy,playing a critical role in helping stomatological students fully understand professional knowledge and humanistic connotations while sub-stantially improving their professional identity.This teaching mode gives a direction for innovative stomatological education,holds sig-nificant importance for cultivating stomatological professionals with both clinical skills and humanistic literacy,possessing substantial potential for promotion,application,and further refinement.

Objective:To analyze the incidence of pregnancy complications and maternal-neonatal outcomes in women with a history of preconception metabolic and bariatric surgery (MBS).Methods:This study was a retrospective cohort study. Pregnant women with singleton pregnancy who delivered in Nanjing Drum Tower Hospital, the Affiliated Hospital of Nanjing University Medical School, from September 2019 to December 2024 were selected as the observation subjects. After propensity score matching, 42 women in the MBS group and 157 women in the control group were finally included. The general clinical characteristics, pregnancy status and maternal-neonatal outcomes of the two groups were compared and analyzed.Results:(1) There were no statistically significant differences in the age, proportion of preconception obesity, chronic hypertension, preconception diabetes and primipara between the MBS group and the control group (all P>0.05). The median interval between surgery and pregnancy of pregnant women in the MBS group was 14.0 months (6.0, 27.5 months). Twenty-nine pregnant women (69%, 29/42) were pregnant after 1 year of surgery, and 13 pregnant women (31%, 13/42) were pregnant within 1 year. (2) The levels of hemoglobin, serum iron and triglyceride in the MBS group were significantly lower than those in the control group in the second and third trimester (all P<0.05), but there were no statistically significant differences in the levels of low-density lipoprotein cholesterol, high-density lipoprotein cholesterol, total cholesterol and albumin between the two groups (all P>0.05). (3) Compared with the control group, the incidence of gestational diabetes mellitus in MBS group [21.7% (34/157) vs 7.1% (3/42)] and the proportion of large for gestational age [23.6% (37/157) vs 2.4% (1/42)] were lower; the incidence of anemia [6.4% (10/157) vs 33.3% (14/42)], fetal growth restriction [7.0% (11/157) vs 23.8% (10/42)] and small for gestational age [3.8% (6/157) vs 19.0% (8/42)] were higher; the differences were statistically significant (all P<0.05). There were no significant differences in the cesarean section rate, premature rupture of membranes rate, postpartum hemorrhage ≥1 000 ml rate, gestational age at delivery and preterm birth rate between the two groups (all P>0.05). The neonatal birth weight of the MBS group was significantly lower than that of the control group [(3 044±523) vs (3 256±491) g, P=0.016], but the proportion of neonates with 1-minute Apgar score<7 and the rate of neonatal intensive care unit admission were not statistically significant (all P>0.05). Conclusions:Women who got pregnant after MBS had lower neonatal weight, decreased incidence of gestational diabetes mellitus and large for gestational age, but higher incidence of small for gestational age and anemia in late pregnancy. It is necessary to focus on the nutritional management of pregnant women with MBS before pregnancy, improve anemia, and strengthen the ultrasound follow-up of fetal growth to optimize the perinatal outcome.

To evaluate the immunogenicity and immunoprotective effects of a tRNA thiouridylase TgMnmA deletion strain of Toxoplasma gondii(T.gondii)on ICR mice,we constructed a mouse model immunized with RH△MnmA.Mice were immunized with 10 RH△MnmA tachyzoites by in-traperitoneal injection.After 30 d,indirect ELISA was used to detect the specific IgG antibody and its subtypes of immunized mice.Spleen lymphocyte suspension was prepared,and the splenic lym-phocyte subsets were analyzed by flow cytometry.Moreover,the relative expression level of cyto-kine mRNA was detected by real-time fluorescence quantitative PCR.After 30 d of immunization,mice were intraperitoneally inoculated with RH△ku80 tachyzoites.At 5 d post infection,the para-site load in the ascites,heart,liver and brain of mice was measured,and the survival of mice within 30 d after infection was observed and recorded.The results showed that compared with the control PBS group,RH△MnmA immunized group produced higher level of IgG and IgG2a antibodies,higher mRNA relative expression level of cytokines IL-2,IL-4,IL-6,IL-10,IL-12 and IFN-γ,and the number of CD4+and CD8a+in spleen lymphocytes also increased significantly.Mean-while,for the attack of RH△ku80 strain,the immune group can effectively reduce the parasite load in the ascites and some tissues,inhibit the reproduction of parasites In vivo,and significantly improve the survival rate of mice.The results of this study showed that the TgMnmA deletion strain of T.gondii can induce strong humoral and cellular immune responses in mice,and provide good immune protection against the infection of RHΔku80 strain,which has the potential to be-come a potentially promising live attenuated vaccine candidate against T.gondii.