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 observe the clinical effect of CHEN Yinglong 's ziwu reinforcing and reducing acupuncture method on post-stroke swallowing dysfunction (PSD) in the pharyngeal phase, and explore its biomechanical mechanism. METHODS: A total of 72 patients with PSD in the pharyngeal phase were randomly divided into a ziwu reinforcing and reducing acupuncture group (36 cases, 2 cases dropped out and 1 case was eliminated) and a conventional acupuncture group (36 cases, 1 case dropped out and 1 case was eliminated). The patients in the conventional acupuncture group were treated with conventional acupuncture at Lianquan (CV 23) and bilateral Fengchi (GB 20), Wangu (GB 12), Tongli (HT 5), Zhaohai (KI 6), Hegu (LI 4), Taichong (LR 3). The patients in the ziwu reinforcing and reducing acupuncture group were treated with CHEN Yinglong 's ziwu reinforcing and reducing acupuncture method on the basis of the conventional acupuncture group's acupoints. Both groups were treated with acupuncture for 30 min each time, once a day, and rested for 1 day after 6 days of treatment, and the treatment lasted 4 weeks. Before and after treatment, the standardized swallowing assessment (SSA) score, Rosenbek penetration aspiration scale (PAS) grade, the shortening rate of mandibular-hyoid distance, swallowing quality of life (SWAL-QOL) score were evaluated in the two groups. The removal rate of nasogastric feeding tube was compared in the two groups, and the clinical efficacy and safety of the two groups were compared. RESULTS: After treatment, the SSA scores and PAS grade of the two groups were lower than those before treatment (P<0.001), and above indexes in the ziwu reinforcing and reducing acupuncture group were lower than those in the conventional acupuncture group (P<0.01, P<0.05). After treatment, the shortening rates of mandibular-hyoid distance and SWAL-QOL scores in the two groups were higher than those before treatment (P<0.001), and the shortening rate of mandibular-hyoid distance in the ziwu reinforcing and reducing acupuncture group was higher than that in the conventional acupuncture group (P<0.001). The removal rate of nasogastric feeding tube in the ziwu reinforcing and reducing acupuncture group was 87.9% (29/33), which was higher than 61.8% (21/34) in the conventional acupuncture group (P<0.05). The total effective rate of the ziwu reinforcing and reducing acupuncture group was 84.8% (28/33), which was higher than 61.8% (21/34) of the conventional acupuncture group (P<0.05). Neither group experienced serious adverse reactions. CONCLUSION CHEN Yinglong 's ziwu reinforcing and reducing acupuncture method can effectively improve swallowing dysfunction in the pharyngeal phase after stroke, reduce the risk of aspiration, and improve quality of life. Its therapeutic effect may be related to increasing the displacement amplitude of the hyoid bone.
Humans ; Male ; Female ; Middle Aged ; Aged ; Acupuncture Therapy ; Stroke/therapy* ; Acupuncture Points ; Deglutition Disorders/physiopathology* ; Deglutition ; Treatment Outcome ; Adult ; Pharynx/physiopathology*

ObjectiveTo investigate the mechanism of Baihuan Xiaoyao Decoction （Xiaoyaosan added with Lilii Bulbus and Albiziae Cortex） in alleviating depression-like behaviors of juvenile rats by regulating the polarization of microglia. MethodSixty juvenile SD rats were randomized into normal control， model， fluoxetine， and low-， medium-， and high-dose （5.36， 10.71， 21.42 g·kg^-1， respectively） Baihuan Xiaoyao decoction groups. The rat model of juvenile depression was established by chronic unpredictable mild stress （CUMS）. The sucrose preference test （SPT） was carried out to examine the sucrose preference of rats. Forced swimming test （FST） was carried out to measure the immobility time of rats. The open field test （OFT） was conducted to measure the total distance， the central distance， the number of horizontal crossings， and the frequency of rearing. Morris water maze （MWM） was used to measure the escape latency and the number of crossing the platform. The immunofluorescence assay was employed to detect the expression of inducible nitric oxide synthase （iNOS， the polarization marker of M1 microglia） and CD206 （the polarization marker of M2 microglia）. Real-time polymerase chain reaction was employed to determine the mRNA levels of iNOS， CD206， pro-inflammatory cytokines ［tumor necrosis factor （TNF）-α， interleukin （IL）-1β， and IL-6］ and anti-inflammatory cytokines （IL-4 and IL-10） in the hippocampus. Western blotting was employed to determine the protein levels of iNOS and CD206 in the hippocampus. The levels of IL-4 and IL-6 in the hippocampus were detected by enzyme-linked immunosorbent assay. ResultCompared with the normal control group， the model rats showed a reduction in sucrose preference （P<0.05）， an increase in immobility time （P<0.05）， decreased motor and exploratory behaviors （P<0.05）， and weakened learning and spatial memory （P<0.05）. In addition， the model rats showed up-regulated mRNA and protein levels of iNOS and mRNA levels of IL-1β， IL-6， and TNF-α （P<0.05）. Compared with the model group， Baihuan Xiaoyao decoction increased the sucrose preference value （P<0.05）， shortened the immobility time （P<0.01）， increased the motor and exploratory behaviors （P<0.05）， and improved the learning and spatial memory （P<0.01）. Furthermore， the decoction down-regulated the positive expression and protein level of iNOS， lowered the levels of TNF-α， IL-1β， and IL-6 （P<0.01）， promoted the positive expression of CD206， and elevated the levels of IL-4 and IL-10 （P<0.01） in the hippocampus of the high dose group. Moreover， the high-dose Baihuan Xiaoyao decoction group had higher sucrose preference value （P<0.01）， shorter immobility time （P<0.01）， longer central distance （P<0.01）， stronger learning and spatial memory （P<0.01）， higher positive expression and protein level of iNOS （P<0.01）， lower levels of TNF-α， IL-1β， and IL-6 （P<0.05， P<0.01）， lower positive expression and mRNA level of iNOS （P<0.05）， and higher levels of IL-4 and IL-10 （P<0.05， P<0.01） than the fluoxetine group. ConclusionBaihuan Xiaoyao decoction can improve the depression-like behavior of juvenile rats by inhibiting the M1 polarization and promoting the M2 polarization of microglia in the hippocampus.

Objective To investigate the correlation between the ratio of lesion volume to uterine volume(T/U),the expression levels of Ki-67 and p16 proteins in lesion tissue,and the recurrence risk of endometrial cancer.Methods A total of 150 patients diagnosed with endometrial carcinoma through pathological examination at Qiandongnan Prefecture People's Hospital were enrolled for follow-up observation.Among them,28 patients experienced recurrence after a 2-year follow-up period,while 122 patients remained recurrence-free.The expression differences of Ki-67 protein and p16 protein in T/U and lesion tissues during surgery were compared between the two groups.Furthermore,these indexes were analyzed based on different pathological features,and the variation in relapse-free survival time was assessed among patients with distinct T/U status as well as Ki-67 and p16 protein expressions.Results The T/U value and the positive expression rate of Ki-67 protein were significantly higher in the relapsed group compared to the non-relapsed group,while the positive expression rate of p16 protein was significantly lower in the relapsed group(P＜0.05).Additionally,patients with T/U ≥ 0.18 had a significantly higher proportion of stage Ⅲ patients and patients with low histological differentiation compared to those with T/U＜0.18(P＜0.05).Furthermore,patients with positive expression of Ki-67 protein exhibited a significantly higher proportion of stage Ⅲ patients,patients with low histological differentiation,and lymph node metastasis compared to those with negative expression of Ki-67 protein(P＜0.05).The proportion of stage Ⅲ patients exhibiting positive p16 protein expression,low histological differentiation,and lymph node metastasis was significantly lower compared to those with negative p16 protein expression(P＜0.05).Patients with endometrial cancer having a T/U ≥0.18 experienced shorter recurrence-free survival time 2 years post-surgery in comparison to patients with T/U＜0.18(x2=6.962,P=0.008).Patients displaying positive Ki-67 expression had a shorter recurrence-free survival time 2 years after surgery than those with negative Ki-67 expression(x2=4.815,P=0.028).The recurrence-free survival time 2 years after surgery for patients expressing p 16 protein positively was longer than that for patients ex-pressing it negatively(x2=4.279,P=0.039).The presence of FIGO stage Ⅲ,lymph node metastasis,depth of myographic invasion ≥1/2,T/U value ≥ 0.18,and positive expression of Ki-67 protein were identified as significant risk factors for postoperative recurrence in endometrial cancer(P＜0.05).Conversely,the positive expression of p16 protein was found to be a protective factor against recurrence in endometrial carcinoma following surgery(P＜0.05).Conclusion The expression of T/U,Ki-67 protein,and p16 protein in endometrial cancer patients is as-sociated with tumor progression and may augment the risk of postoperative recurrence.

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.

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.