Mannheimia varigena(M.varigena)is a significant pathogen causing bovine respiratory disease.Transferrin binding protein B(TbpB)is a lipoprotein directly exposed to the outer mem-brane of the cell,which is not only involved in the bacterial iron metabolism pathway,but also an important virulence factor.This study aims to lay the groundwork for developing novel subunit vaccines by conducting site-directed mutagenesis on M.varigena TbpB binding-related residues and evaluating their immunogenicity.Based on whole-genome sequencing of a bovine M.varigena i-solate,its iron uptake pathway was predicted.Key amino acid residues of M.varigena TbpB that play a role for binding to bovine transferrin(bTf)was identified by bioinformatics.We constructed two M.varigena TbpB mutants(Y205A and Y258A)and assessed their bTf binding activity and immunogenicity through dot blot assays and mouse immunization studies.Dot blot assays result showed that the Y258A mutation caused TbpB to lose its ability to bind to bTf.Mouse immuniza-tion studies showed that,compared to wild-type TbpB,the mutants of TbpB induced higher levels of specific antibodies.In challenge experiments,mice immunized with mutant TbpB exhibited high-er survival rates.These results demonstrate that site-directed mutagenesis can enhance the immu-nogenicity of TbpB.This study provides a novel approach for developing new subunit vaccines a-gainst M.varigena.

Objective To explore the therapeutic effect of precise disconnection of pargastric varices guided by endoscopic ultrasound in the treatment of esophagogastric variceal bleeding.Method A retrospective analysis was conducted on 20 patients with cirrhosis esophagogastric variceal bleeding treated with endoscopic ultrasound-guided precise disconnection of pargastric varices from January 1,2024 to December 31,2024.The efficacy was analyzed.Result All 20 patients successfully completed the precise disconnection of pargastric varices under the guidance of endoscopic ultrasound.The injection of tissue gel combined with the placement of spring coils(14 cases)and the injection of tissue gel alone(4 cases)successfully blocked the pargastric varices.All patients did not experience perforation,esophageal and cardia stenosis,massive bleeding,septicemia,or ectopic embolization.One patient who received tissue gel alone had slight bleeding from the pargastric varices after surgery and improved after 3 days of treatment to reduce portal vein pressure.Another one patient who received tissue gel alone had a low-grade fever and normal body temperature after 3 days of anti-infection treatment.Conclusion Precise disconnection of pargastric varices under the guidance of endoscopic ultrasound has a good therapeutic effect on esophagogastric variceal bleeding,with fewer complications such as ectopic embolization,massive bleeding,infection,and perforation.However,close follow-up observation is still needed to address the issue of pargastric varices.

Mannheimia varigena(M.varigena)is a significant pathogen causing bovine respiratory disease.Transferrin binding protein B(TbpB)is a lipoprotein directly exposed to the outer mem-brane of the cell,which is not only involved in the bacterial iron metabolism pathway,but also an important virulence factor.This study aims to lay the groundwork for developing novel subunit vaccines by conducting site-directed mutagenesis on M.varigena TbpB binding-related residues and evaluating their immunogenicity.Based on whole-genome sequencing of a bovine M.varigena i-solate,its iron uptake pathway was predicted.Key amino acid residues of M.varigena TbpB that play a role for binding to bovine transferrin(bTf)was identified by bioinformatics.We constructed two M.varigena TbpB mutants(Y205A and Y258A)and assessed their bTf binding activity and immunogenicity through dot blot assays and mouse immunization studies.Dot blot assays result showed that the Y258A mutation caused TbpB to lose its ability to bind to bTf.Mouse immuniza-tion studies showed that,compared to wild-type TbpB,the mutants of TbpB induced higher levels of specific antibodies.In challenge experiments,mice immunized with mutant TbpB exhibited high-er survival rates.These results demonstrate that site-directed mutagenesis can enhance the immu-nogenicity of TbpB.This study provides a novel approach for developing new subunit vaccines a-gainst M.varigena.

Objective To explore the therapeutic effect of precise disconnection of pargastric varices guided by endoscopic ultrasound in the treatment of esophagogastric variceal bleeding.Method A retrospective analysis was conducted on 20 patients with cirrhosis esophagogastric variceal bleeding treated with endoscopic ultrasound-guided precise disconnection of pargastric varices from January 1,2024 to December 31,2024.The efficacy was analyzed.Result All 20 patients successfully completed the precise disconnection of pargastric varices under the guidance of endoscopic ultrasound.The injection of tissue gel combined with the placement of spring coils(14 cases)and the injection of tissue gel alone(4 cases)successfully blocked the pargastric varices.All patients did not experience perforation,esophageal and cardia stenosis,massive bleeding,septicemia,or ectopic embolization.One patient who received tissue gel alone had slight bleeding from the pargastric varices after surgery and improved after 3 days of treatment to reduce portal vein pressure.Another one patient who received tissue gel alone had a low-grade fever and normal body temperature after 3 days of anti-infection treatment.Conclusion Precise disconnection of pargastric varices under the guidance of endoscopic ultrasound has a good therapeutic effect on esophagogastric variceal bleeding,with fewer complications such as ectopic embolization,massive bleeding,infection,and perforation.However,close follow-up observation is still needed to address the issue of pargastric varices.

Objective:To compare and analyze the differences in clinical and pathological features of uterine smooth muscle tumor of uncertain malignant potential (STUMP), common uterine leiomyoma (UL), and cellular uterine leiomyoma (CUL), and to identify biomarkers for predicting STUMP recurrence.Methods:A total of 72 cases of STUMP patients (STUMP group) treated at the First Affiliated Hospital of Nanjing Medical University and the First Affiliated Hospital of Soochow University were collected from June 2015 to March 2024. Additionally, 72 cases of UL and 72 cases of CUL (UL group and CUL group) in the same period were collected as controls. The clinical and pathological features of the three groups were compared, and the recurrence rates and related factors affecting STUMP recurrence were analyzed.Results:(1) Comparison of clinical and pathological features: there were statistically significant differences in age, history of myomectomy, and preoperative serum lactate dehydrogenase (LDH) levels among STUMP, UL, and CUL groups (all P<0.05). STUMP group were significantly older than UL group ( P<0.05). The proportions of STUMP group with a history of myomectomy and elevated preoperative serum LDH levels were significantly higher than those in UL and CUL groups (all P<0.05). On ultrasound, 16 cases of STUMP patients (22%, 16/72), 2 cases of UL patients (3%, 2/72), and 8 cases of CUL patients (11%, 18/72) had unclear fibroid borders, with significant differences between the three groups ( χ2=12.94, P=0.002), with STUMP group significantly higher than UL group ( P<0.05). Regarding immunohistochemistry, the proportion of p16 positivity, p53 mutations, and nuclear antigen associated with cell proliferation (Ki-67) >10% were significantly higher in STUMP group compared to UL group ( P<0.05). In terms of surgical approach, 52 cases of STUMP patients (72%, 52/72) underwent hysterectomy, compared to 27 cases of UL patients (38%, 27/72) and 38 cases of CUL patients (53%, 38/72), with a significant difference between the three groups ( χ2=17.89, P=0.001). The proportion of patients who underwent myomectomy was significantly lower in STUMP group compared to UL and CUL groups (both P<0.05). Among the 20 cases of STUMP patients who underwent myomectomy, 6 patients had a subsequent total hysterectomy after being diagnosed with STUMP. (2) Comparison of recurrence: the median follow-up time for the STUMP, UL, and CUL groups was 38, 12, and 29 months, respectively. During the follow-up period, 3 cases (6%, 3/53) in STUMP group, 4 cases (7%, 4/55) in UL group, and 8 cases (13%, 8/62) in CUL group had recurrence, with no significant differences between the three groups ( χ2=1.91, P=0.411). Among the 3 cases of STUMP patients with recurrence (in the pelvic cavity, liver, and abdominal wall), 2 cases had STUMP pathology on recurrence, and 1 case progressed to well-differentiated uterine leiomyosarcoma. (3) Related factors affecting STUMP recurrence: when comparing preoperative body mass index, serum LDH levels, and Ki-67 positivity ≥30% between recurrent and non-recurrent patients, significant differences were observed (all P<0.05). Univariate logistic regression analysis showed that Ki-67 positivity ≥30% was a significant risk factor for STUMP recurrence ( OR=24.67, 95% CI: 1.70-357.36, P=0.019). Conclusions:Factors such as age, history of myomectomy, preoperative serum LDH levels, and ultrasound findings of unclear fibroid borders are helpful for distinguishing STUMP from UL and CUL. Elevated preoperative serum LDH levels and Ki-67 positivity ≥30% have predictive value for STUMP recurrence. Active postoperative follow-up is essential, whether STUMP patients undergo myomectomy or hysterectomy.

Objective:To improve the quality standard of Xiaoer Shenzhu Jianpi Pills.Methods:The microscopic identification method was used to qualitatively identify Hordei Fructus Germinatus.TLC was used to qualitatively i-dentify of Atractylodis Macrocephalae Rhizoma,Glycyrrhizae Radix Et Rhizoma,Citri Reticulatae Pericapium.HPLC was adopted to determine the contents of hesperidin and ammonium glycyrrhizinate.Results:The microscop-ic identification of Hordei Fructus Germinatus had strong specificity and high viewing rate.The TLC identification spots of Atractylodis Macrocephalae Rhizoma,Glycyrrhizae Radix Et Rhizoma,Citri Reticulatae Pericapium were clear,with good separation and durability.The linear ranges of hesperidin and ammonium glycyrrhizinate were 0.060 0-1.501 0 μg and 0.020 1-0.502 2 μg,respectively(r≥0.999 8).The average recoveries(n=9)were 99％,96％and RSDs were 2.8％and 1.8％.The RSDs of precision,stability and repeatability of the two components were less than 3.0％.Conclusion:The newly established quality standard increases the quality control research of microscopic identification,thin layer chromatography and content determination.The experimental method is accurate and reliable,simple and fast,and has good applicability.It can be used for the quality control and evaluation of Xiaoer Shenzhu Jianpi Pills.

Background/Aims: Low educational attainment is a well-established risk factor for nonalcoholic fatty liver disease (NAFLD) in developed areas. However, the association between educational attainment and the risk of NAFLD is less clear in China. Methods: A cross-sectional study including over 200,000 Chinese adults across mainland China was conducted. Information on education level and lifestyle factors were obtained through standard questionnaires, while NAFLD and advanced fibrosis were diagnosed using validated formulas. Outcomes included the risk of NAFLD in the general population and high probability of fibrosis among patients with NAFLD. Logistic regression analysis was employed to estimate the risk of NAFLD and fibrosis across education levels. A causal mediation model was used to explore the potential mediators. Results: Comparing with those receiving primary school education, the multi-adjusted odds ratios (95% confidence intervals) for NAFLD were 1.28 (1.16 to 1.41) for men and 0.94 (0.89 to 0.99) for women with college education after accounting for body mass index. When considering waist circumference, the odds ratios (95% CIs) were 0.94 (0.86 to 1.04) for men and 0.88 (0.80 to 0.97) for women, respectively. The proportions mediated by general and central obesity were 51.00% and 68.04% for men, while for women the proportions were 48.58% and 32.58%, respectively. Furthermore, NAFLD patients with lower educational attainment showed an incremental increased risk of advanced fibrosis in both genders. Conclusions In China, a low education level was associated with a higher risk of prevalent NAFLD in women, as well as high probability of fibrosis in both genders.

ObjectiveTo investigate the acute effects of compound air pollution on children’s respiratory function. MethodsUsing panel group study design， 223 students in five classes of grade 4 from two primary schools （a， b） in Xuhui and Hongkou districts of Shanghai were randomly selected to measure pulmonary function and exhaled nitric oxide （FeNO）. The first three tests were carried out from May to June in 2020， and the fourth test was carried out from September to December in 2021. At the same time， the daily and hourly mean values of PM_2.5， PM₁₀， SO₂， NO₂， O₃ and CO was collected from the nearby air quality monitoring points of the two schools during the same period ， as well as meteorological monitoring data （temperature， humidity， wind speed and atmospheric pressure）. The linear mixed effect model was used to analyze the effects of air pollution on pulmonary function and respiratory inflammation in the summer. ResultsThe results of single pollutant model showed that PM_2.5， PM_10， SO₂ and NO₂ were positively correlated with FeNO， and the effect was reflected in lag0， lag1 and lag3 （P<0.05）. PM_2.5， PM₁₀ and NO₂ were negatively correlated with the changes of lung function FEF_25%， FEF_50%， FEF_75%， FeF_25%-75%， PEF， FVC， FEV1 and FEV1/FVC， and the effect was reflected in lag0 to lag3 days （P<0.05）. The results of the dual pollutant model showed that the concentration changes of SO₂ and NO₂ were significantly correlated with the decrease of FEV1 when combined with O₃ or PM_2.5 （P<0.01）， and the concentration changes of PM_2.5 was significantly correlated with the increase of FeNO when O₃， SO₂ and NO₂ were combined respectively （P<0.01）. The effects of the dual pollutant model were greater than the effect of PM_2.5 single pollutant model. ConclusionThe health effects of different air pollutants on children’s respiratory tract function indexes in summer are different. The combined effects of two pollutants on the lung function of children increased to different degrees. Although air pollution is light in summer， it still has an impact on children’s respiratory tract function index and inflammation index， and the combined effect of dual pollutants is more significant than that of single pollutant.

Objective:To explore the establishment of a Cox proportional hazard prediction model for the re-cent(within a week)delivery of pregnant women with threatened early preterm labor(28-33+6 weeks).Meth-ods:The clinical data of 293 patients with threatened early preterm labor at 28-33+6 weeks admitted to the De-partment of Obstetrics,Affiliated Hospital of North Sichuan Medical College from January 2021 to December 2022 were retrospectively collected.According to whether the patient gave birth recently(within a week)during hospi-talization,they were divided into delivery group(n=88)and non-delivery group(n=205).Demographic data,specialty conditions and clinical biochemical indicators were collected,and the differences of each observation in-dex between the two groups were compared.Cox multivariate analysis was used to screen variables,and then the Cox proportional hazard prediction model was constructed to calculate the hazard ratio(HR).The C-index was used to test the predictive ability of the model,and the nomogram was drawn to visualize the Cox proportional hazard prediction model.The calibration curve was used to test the consistency between the model prediction re-sults and the actual situation.Results:The average age of the delivery group was 30.2±5.0 years old,and the average prolonged pregnancy time was 61.3±47.5 hours.Compared to the group that gave birth and did not give birth,the proportion of patients with body mass index(BMI)at admission,parity＞1,gestational diabetes mellitus or diabetic pregnancy combined(GDM/PGDM),abnormal fetal orientation,regular contractions at admis-sion,vaginal cleanliness≥ Ⅲ,white blood cell count ≥10 ×109/L at admission and cervical length＜20 mm,the difference was statistically significant(P＜0.05).Multivariate Cox regression analysis:BMI＞30 kg/m2(HR 2.178,95％CI 1.208-3.928,P=0.010),parity＞1(HR 3.095,95％CI 1.759-5.447,P＜0.001),regular uterine contraction(HR3.447,95％CI 2.216-5.362,P＜0.001),cervical length＜20 mm(HR3.594,95％CI 2.289-5.646,P＜0.001)and white blood cell count≥10 ×109/L(HR2.124,95％CI 1.352-3.335,P=0.001)on admission were independent risk factors for recent(within a week)delivery of pregnant women with threatened early preterm birth.When the above 5 indicators were included in the prediction model,the C-index was 0.797(95％CI 0.750-0.844),indicating that the model had good prediction ability.The calibration chart shows that the prediction results of the model are in good agreement with the actual results.Conclusions:Preg-nant women with BMI＞30 kg/m2 at admission,parity＞1,regular uterine contraction at admission,cervical length＜20 mm at admission and white blood cell count≥10 ×109/L at admission are prone to premature delivery in the near future.Cox proportional risk prediction model based on the above risk factors has certain accuracy,which can be used to guide clinical workers to take corresponding intervention measures in advance to avoid or reduce the possibility of premature delivery in the near future and improve maternal and infant outcomes.