To verify whether chaperones can promote the soluble expression of PlpE in Escherichia coli and whether the expressed protein is active,prokaryotic expression and Western blot detection were performed.The results showed that:The PlpE prokaryotic expression vector pET-32a(+)-plpE was expressed as inclusion body,and the expression form was not changed by changing the concentration of inducer,induction time and temperature.The companion proteins pG-KJE8,pGro7,pKJE7 and pG-Tf2 were co-expressed with pET-32a(+)-plpE in Eschierichia coli expres-sion system,respectively.When the final concentration of IPTG of 0.5 mmol/L,L-arabinose of 0.5 g/L or tetracycline of 5.0 μg/L were added as inducers and induced at 37 ℃ for 8 h,the results showed that the molecular companion pGro7 could change the expression of rp-PlpE from inclu-sion body to soluble expression.pG-KJE8,pKJE7 and pG-Tf2 had no effect on the expression of rp-PlpE.The soluble rp-PlpE can react specifically with the positive serum of Mannheimia haemolyti-ca.Therefore,the study showed that the co-expression of the chaperone protein pGro7 can make the rp-PlpE protein express in a soluble form,and the purified protein exhibits reactogenicity.These findings lay the foundation for the establishment of a subunit vaccine and serological diagno-sis methods for Mannheimia haemolytica.

To verify whether chaperones can promote the soluble expression of PlpE in Escherichia coli and whether the expressed protein is active,prokaryotic expression and Western blot detection were performed.The results showed that:The PlpE prokaryotic expression vector pET-32a(+)-plpE was expressed as inclusion body,and the expression form was not changed by changing the concentration of inducer,induction time and temperature.The companion proteins pG-KJE8,pGro7,pKJE7 and pG-Tf2 were co-expressed with pET-32a(+)-plpE in Eschierichia coli expres-sion system,respectively.When the final concentration of IPTG of 0.5 mmol/L,L-arabinose of 0.5 g/L or tetracycline of 5.0 μg/L were added as inducers and induced at 37 ℃ for 8 h,the results showed that the molecular companion pGro7 could change the expression of rp-PlpE from inclu-sion body to soluble expression.pG-KJE8,pKJE7 and pG-Tf2 had no effect on the expression of rp-PlpE.The soluble rp-PlpE can react specifically with the positive serum of Mannheimia haemolyti-ca.Therefore,the study showed that the co-expression of the chaperone protein pGro7 can make the rp-PlpE protein express in a soluble form,and the purified protein exhibits reactogenicity.These findings lay the foundation for the establishment of a subunit vaccine and serological diagno-sis methods for Mannheimia haemolytica.

Objective:To explore the relationship between childhood obesity, gut microbiota, and short chain fatty acids(SCFAs) based on 16S rRNA sequencing.Methods:A total of 162 obese children admitted to our hospital and 162 age-matched healthy children with normal body weight from the same period were selected as the control group. Baseline data were collected. Gut microbiota composition and abundance were analyzed using 16S rRNA sequencing, and fecal SCFA levels were measured by gas chromatography-mass spectrometry. Then, multiple linear regression, multivariable logistic regression, Spearman correlation, and restricted cubic spline models were applied to explore the relationships among baseline data, childhood obesity, gut microbiota, and their metabolites. Results:The α diversity(ACE, Chao1, Shannon index) of intestinal flora in the obese group was significantly lower than that in the control group( P<0.05). The abundance of Firmicutes, Actinobacteria, Acinetobacter, Ruminococcus, Clostridium, and Prevotella increased significantly, while the abundance of Bacteroidetes, Bifidobacterium, Akkermansia, Coprococcus, Streptobacter, and Eubacterium decreased significantly( P<0.05). The levels of butyrate, propionate, acetate, and total SCFAs in the obese group decreased significantly( P<0.05). Multivariate linear regression analysis showed that parental obesity and snack consumption frequency were positively related to the abundance of Firmicutes and Actinobacteria in children′s gut, but negatively related to the abundance of Proteobacteria. As for Bacteroidetes, butyrate and propionate levels, they were negatively linked to exercise time, eating rate, and birth weight, and also positively related to Firmicutes. The results of correlation analysis showed that Firmicutes, Actinobacteria, Anaerostipes, Acinetobacter, Ruminococcus, Clostridium, and Prevotella were significantly positively correlated with childhood obesity, while Bacteroidetes, Bifidobacterium, Akkermansia, butyrate, propionate, acetate, and total SCFAs were significantly negatively correlated with childhood obesity. Nonlinear analysis showed that butyrate, propionate, acetate, and Firmicutes/ Bacteroidetes were nonlinearly correlated with the occurrence of obesity( P<0.05). Conclusion:Childhood obesity is closely related to reduced gut microbiota diversity, altered abundances of specific microbial taxa, and reduced levels of SCFA. Modulating gut microbiota may provide novel microbial targets for obesity prevention and treatment.

Objective:To explore the relationship between childhood obesity, gut microbiota, and short chain fatty acids(SCFAs) based on 16S rRNA sequencing.Methods:A total of 162 obese children admitted to our hospital and 162 age-matched healthy children with normal body weight from the same period were selected as the control group. Baseline data were collected. Gut microbiota composition and abundance were analyzed using 16S rRNA sequencing, and fecal SCFA levels were measured by gas chromatography-mass spectrometry. Then, multiple linear regression, multivariable logistic regression, Spearman correlation, and restricted cubic spline models were applied to explore the relationships among baseline data, childhood obesity, gut microbiota, and their metabolites. Results:The α diversity(ACE, Chao1, Shannon index) of intestinal flora in the obese group was significantly lower than that in the control group( P<0.05). The abundance of Firmicutes, Actinobacteria, Acinetobacter, Ruminococcus, Clostridium, and Prevotella increased significantly, while the abundance of Bacteroidetes, Bifidobacterium, Akkermansia, Coprococcus, Streptobacter, and Eubacterium decreased significantly( P<0.05). The levels of butyrate, propionate, acetate, and total SCFAs in the obese group decreased significantly( P<0.05). Multivariate linear regression analysis showed that parental obesity and snack consumption frequency were positively related to the abundance of Firmicutes and Actinobacteria in children′s gut, but negatively related to the abundance of Proteobacteria. As for Bacteroidetes, butyrate and propionate levels, they were negatively linked to exercise time, eating rate, and birth weight, and also positively related to Firmicutes. The results of correlation analysis showed that Firmicutes, Actinobacteria, Anaerostipes, Acinetobacter, Ruminococcus, Clostridium, and Prevotella were significantly positively correlated with childhood obesity, while Bacteroidetes, Bifidobacterium, Akkermansia, butyrate, propionate, acetate, and total SCFAs were significantly negatively correlated with childhood obesity. Nonlinear analysis showed that butyrate, propionate, acetate, and Firmicutes/ Bacteroidetes were nonlinearly correlated with the occurrence of obesity( P<0.05). Conclusion:Childhood obesity is closely related to reduced gut microbiota diversity, altered abundances of specific microbial taxa, and reduced levels of SCFA. Modulating gut microbiota may provide novel microbial targets for obesity prevention and treatment.

Environmental toxicants have been linked to aging and age-related diseases. The emerging evidence has shown that the enhancement of detoxification gene expression is a common transcriptome marker of long-lived mice, Drosophila melanogaster, and Caenorhabditis elegans. Meanwhile, the resistance to toxicants was increased in long-lived animals. Here, we show that farnesoid X receptor (FXR) agonist obeticholic acid (OCA), a marketed drug for the treatment of cholestasis, may extend the lifespan and healthspan both in C. elegans and chemical-induced early senescent mice. Furthermore, OCA increased the resistance of worms to toxicants and activated the expression of detoxification genes in both mice and C. elegans. The longevity effects of OCA were attenuated in Fxr ^-/- mice and Fxr homologous nhr-8 and daf-12 mutant C. elegans. In addition, metabolome analysis revealed that OCA increased the endogenous agonist levels of the pregnane X receptor (PXR), a major nuclear receptor for detoxification regulation, in the liver of mice. Together, our findings suggest that OCA has the potential to lengthen lifespan and healthspan by activating nuclear receptor-mediated detoxification functions, thus, targeting FXR may offer to promote longevity.

Plant-derived nanovesicles have gained attention given their similarity to mammalian exosomes and advantages such as low cost,sustainability,and tissue targeting.Thus,they hold promise for disease treatment and drug delivery.In this study,we proposed a time-efficient method,PEG 8000 combined with sucrose density gradient centrifugation to prepare ginger-derived nanovesicles(GDNVs).Subse-quently,curcumin(CUR)was loaded onto GDNV by ultrasonic incubation.The optimum conditions for ginger-derived nanovesicles loaded with curcumin(CG)were ultrasound time of 3 min,a carrier-to-drug ratio(GDNV:CUR)of 1∶1.The study achieved a high loading capacity(94.027％±0.094％)and encapsu-lation efficiency(89.300％±0.344％).Finally,the drugs'in vivo distribution and anti-colitis activity were investigated in mice.CG was primarily distributed in the colon after oral administration.Compared to CUR and GDNV,CG was superior in improving disease activity,colon length,liver and spleen coefficients,myeloperoxidase activity,and biochemical factor levels in ulcerative colitis(UC)mice.In addition,CG plays a protective role against UC by modulating serum metabolite levels and gut flora.In summary,our study demonstrated that GDNV can be used for CUR delivery with enhanced therapeutic potential.

Objective To compare and analyze the efficacy and risks of percutaneous CT/ultrasound-guided bipolar radiofrequency ablation(RFA)under local anesthesia with robotic-assisted laparoscopic partial nephrectomy(RAPN)for the treatment of sporadic small renal mass.Methods A retrospective study was conducted on 93 consecutive patients with T1a stage small renal mass during Mar.2019 and Oct.2021.Among them,51 underwent RAPN,and 42 underwent RFA.General information,tumor characteristics,perioperative and follow-up data were collected and statistically analyzed.Results There were no significant differences in general information and tumor characteristics between the two groups(P＞0.05).The operation time[(96.0±20.0)min vs.(113.5±24.1)min,P＜0.001],hospital stay[(3.5±0.8)day vs.(6.9±1.8)day,P＜0.001],and hospital costs[(2.4±0.7)ten thousand yuan vs.(6.6±0.4)ten thousand yuan,P＜0.001]were significantly decreased in the RFA group than in the RAPN group.There were no significant differences in the incidence of perioperative complications and long-term disease-free survival rate between the two groups(P＞0.05).However,the difference between one-year postoperative estimated glomerular filtration rate(eGFR)and preoperative eGFR was significantly lower in the RFA group than in the RAPN group[-2.3(-4.7-1.3)mL/(min·1.73 m2)vs.-5.0(-9.1 2.8)mL/(min·1.73 m2),P=0.003],and the reduction of one-year postoperative creatinine and preoperative creatinine was slightly lower in the RFA group than in the RAPN group[4.0(-0.2-5.5)μmol/L vs.4.5(1.8-9.2)μmol/L,P=0.122].Conclusion RFA can achieve comparable disease-free survival rate as RAPN in the treatment of T1a renal tumor,and can effectively preserve renal function,reduce medical costs,save medical resources,and lower the incidence of perioperative complications.

Objective:To explore the effectiveness and safety of percutaneous radiofrequency ablation for renal tumors, guided by both ultrasound and CT, under local anesthesia.Methods:A retrospective analysis was conducted on the clinical data of 40 patients with renal tumors admitted to Nanjing Drum Tower Hospital between January 2018 and December 2022. This treatment involved ultrasound/CT dual-guided radiofrequency ablation under local anesthesia. The cohort included 33 males and 7 females, with an average age of (61.5±11.9) years old and a body mass index (BMI) of (24.79±3.37) kg/m 2. The tumors were located in the left kidney in 20 cases and the right kidney in 16 cases, with 4 cases involving bilateral renal tumors. There were 44 tumors in 40 patients, with the maximum tumor diameter ranging from 1.0 to 4.0 cm [mean (2.3 ± 0.7) cm]. Distribution by kidney pole was as follows: 15 cases at the upper pole, 21 at the middle pole, and 8 at the lower pole. Of the tumors, 23 were exophytic, 5 were endophytic, and 16 exhibited mixed features. There were 2 patients with multiple metastases before surgery(including 1 patient with bilateral renal tumor). Preoperative serum creatinine level was 68.0(56.5, 87.5)μmol/L, and the estimated glomerular filtration rate (eGFR) was 114.2 (79.6, 132.4) ml/(min·1.73 m 2). All patients underwent renal biopsy before or during radiofrequency ablation. Percutaneous radiofrequency ablation surgery was performed using ultrasound and CT dual guidance on all patients, ensuring complete tumor destruction during the procedure as confirmed by dual positioning. Patients with bilateral tumors underwent two separate surgeries, spaced one month apart. Postoperatively, closely monitor the patient's vital signs and conduct long-term follow-ups to record any recurrence and metastasis. Results:In this series, all 40 procedures (involving 44 renal units) were successfully completed under local anesthesia without any need for blood transfusion, conversion to open surgery, or perioperative deaths. The average radiofrequency ablation time was (9.5 ± 3.6) min. Tumor characteristics included predominantly exophytic growths (23 cases, 52.3%), with 31 cases (70.5%) located more than 7 mm from the collecting system and 28 cases (63.6%) positioned posteriorly. Thirteen cases (29.5%) were entirely outside the polar line. The average R. E.N.A.L. nephrometry score was 6.1±0.2. Pathological examination revealed 34 cases of clear cell carcinoma, 2 of papillary renal cell carcinoma, 4 of unclassified renal cell carcinoma, and 4 benign renal tumors. In this cohort, two patients with bilateral renal tumors exhibited benign tumors on one side, while two other patients had malignant tumors in both kidneys. All 40 malignant tumors identified in 38 cases were classified at stage cT 1a.Postoperative serum creatinine level was 71.5 (59.0, 94.3) μmol/L, showing no statistically significant change from preoperative levels ( P > 0.05). Similarly, the eGFR post-operation was 107.4 (79.7, 132.2) ml/(min·1.73 m 2), which also did not differ significantly from preoperative values ( P > 0.05). There were no postoperative complications of Clavien-Dindo grade ≥Ⅱ, except for one case of severe pain (score 7-10). The follow-up period ranged from 15 to 70 months. Among the 38 cases, 36 patients did not have distant metastasis before surgery. There were 2 patients (5.5%) with local recurrence within 60 months after surgery. Among them, one case relapsed 6 months after radiofrequency ablation and was treated with partial nephrectomy. The patient was followed up for 60 months after the second treatment, and no local recurrence occurred. Another patient relapsed 41 months after surgery and was treated with radiofrequency ablation again. The patient was followed up for 12 months after the second treatment, and no local recurrence occurred. Two patients with distant metastasis before surgery were treated with targeted therapy plus immunotherapy for 12 months after surgery. One case had local recurrence 8 months after surgery and was treated with partial nephrectomy. The patient was followed up for 60 months after partial nephrectomy and no local recurrence occurred. Another patient with bilateral renal tumors developed left kidney recurrence 34 months after radiofrequency ablation, and underwent left partial nephrectomy. The right kidney recurred 42 months after radiofrequency ablation and underwent radiofrequency ablation again. After the second right renal radiofrequency ablation, no local recurrence occurred during 12 months of follow-up. Conclusions:Ultrasound/CT dual-guided percutaneous radiofrequency ablation, performed under local anesthesia for treating renal tumors, has minimal impact on the patient's renal function. The procedure boasts a low complication rate, with no postoperative severe complications. Additionally, the postoperative tumor control is effective, making it a safe and minimally invasive surgical option.

Plant-derived nanovesicles have gained attention given their similarity to mammalian exosomes and advantages such as low cost, sustainability, and tissue targeting. Thus, they hold promise for disease treatment and drug delivery. In this study, we proposed a time-efficient method, PEG 8000 combined with sucrose density gradient centrifugation to prepare ginger-derived nanovesicles (GDNVs). Subsequently, curcumin (CUR) was loaded onto GDNV by ultrasonic incubation. The optimum conditions for ginger-derived nanovesicles loaded with curcumin (CG) were ultrasound time of 3 min, a carrier-to-drug ratio (GDNV:CUR) of 1:1. The study achieved a high loading capacity (94.027% ± 0.094%) and encapsulation efficiency (89.300% ± 0.344%). Finally, the drugs' in vivo distribution and anti-colitis activity were investigated in mice. CG was primarily distributed in the colon after oral administration. Compared to CUR and GDNV, CG was superior in improving disease activity, colon length, liver and spleen coefficients, myeloperoxidase activity, and biochemical factor levels in ulcerative colitis (UC) mice. In addition, CG plays a protective role against UC by modulating serum metabolite levels and gut flora. In summary, our study demonstrated that GDNV can be used for CUR delivery with enhanced therapeutic potential.

【Objective】 To evaluate the efficacy of ultrasound-guided transperineal puncture and drainage in the treatment of pelvic lymphatic cyst. 【Methods】 A total of 26 patients with pelvic lymphocele who failed with conservative treatment received transperineal puncture and drainage guided by rectal ultrasound. 【Results】 All operations were successful without serious complications. The symptoms in 24 patients relieved within 48 hours after catheter drainage, and 2 patients had the catheter removed after continuous drainage for 3 weeks. Of the 26 patients, 19(73.1%) were cured and 7(26.9%) were relieved. The total effective rate was 100%. 【Conclusion】 Transperineal drainage guided by rectal ultrasound is a safe and effective treatment for pelvic lymphocele.