ObjectiveTo optimize the extraction method for polysaccharides from turnip（Brassica rapa）， and analyze and evaluate the primary structure of the isolated and purified turnip polysaccharide fraction（BP-1） and its hypoglycemic effects in diabetic zebrafish. MethodsTaking polysaccharide yield as the evaluation index， a semi-bionic extraction method was employed. Single-factor experiments and Box-Behnken response surface methodology were used to investigate three factors of solid-to-liquid ratio， extraction time and extraction temperature， in order to optimize the extraction process. BP-1 was isolated and purified using the Sevage method and DEAE-52 cellulose column chromatography. Structural characterization of the turnip polysaccharides was performed using ultraviolet-visible spectrophotometry（UV）， gas chromatography-mass spectrometry（GC-MS）， Congo red assay， and Fourier-transform infrared spectroscopy（FT-IR） to determine purity， monosaccharide composition， triple-helix structure， and functional groups. The microstructure of the polysaccharides was observed using scanning electron microscopy（SEM） and atomic force microscopy（AFM）. Zebrafish were divided into the blank group（adding E3 medium）， and BP-1-1， BP-1-10， BP-1-50， BP-1-200， BP-1-1 000 groups（adding BP-1 solutions at concentrations of 1， 10， 50， 200， 1 000 mg·L^-1， respectively）， and zebrafish embryos were subjected to a 96-hour exposure experiment. The maximum tolerated concentration of BP-1 in zebrafish was determined by evaluating its effects on phenotype， survival rate， malformation rate， and heart rate. Experimental animals were randomly divided into the blank group， model group， BP-1-10 group（10 mg·L^-1）， BP-1-50 group（50 mg·L^-1）， and BP-1-200 group（200 mg·L^-1）. The blank group was cultured in E3 medium， the model and treatment groups were induced to establish a diabetic model in 4 day-post-fertilization（dpf） zebrafish embryos using 10 g·L^-1 of glucose combined with 500 µmol·L^-1 of alloxan. The treatment groups received corresponding doses of BP-1 solution， while the blank and model groups received an equal volume of saline. Glucose and insulin（INS） levels were measured using enzyme-linked immunosorbent assay（ELISA） kits， the effects on the liver were observed by hematoxylin-eosin（HE） histopathological sections. The mRNA expression levels of glucagon（Glucagon）， insulin（Insa）， and phosphoenolpyruvate carboxykinase 1（PCK1） were detected with real-time fluorescence quantitative polymerase chain reaction（Real-time PCR）. ResultsThe optimized extraction conditions were determined as follows：solid-to-liquid ratio of 1∶40（g·mL^-1）， extraction time of 66 min， and extraction temperature of 79 ℃. Under these conditions， the yield of turnip polysaccharides was （10.34±0.96）%. UV analysis indicated that BP-1 contained no proteins or nucleic acids， GC-MS analysis revealed that BP-1 consisted of six monosaccharides（arabinose， rhamnose， ribose， mannose， galactose and glucose）. Congo red assay indicated that the molecular conformation did not exhibit a triple-helix structure， FT-IR analysis showed the presence of α-glycosidic bonds and uronic acids， SEM analysis revealed an irregular flaky structure with a flat and smooth surface， AFM analysis suggested that the aggregated structure might be formed by the entanglement of molecular chains and intramolecular hydrogen bonding. The maximum tolerated concentration of BP-1 in zebrafish over 96 h was determined to be 200 mg·L^-1. Pharmacodynamic results showed that， compared with the blank group， the model group exhibited significantly increased glucose levels and significantly decreased INS levels（P<0.01）. Compared with the model group， the BP-1-50 group significantly reduced glucose levels and increased INS levels（P<0.05）. Histopathological examination of liver tissue revealed that various doses of BP-1 had a certain reparative effect on damaged liver tissue. The liver tissue structure in the BP-1-200 group was nearly normal， with hepatocytes appearing plump. Real-time PCR results showed that， compared with the blank group， the model group exhibited significantly upregulated mRNA expressions of Glucagon and PCK1， and significantly downregulated mRNA expression of Insa（P<0.01）. Compared with the model group， the BP-1-50 and BP-1-200 groups showed significantly downregulated mRNA expressions of Glucagon and PCK1， and significantly upregulated mRNA expression of Insa（P<0.01）. ConclusionThe semi-bionic extraction method for turnip polysaccharides yields a high extraction rate， is simple to operate， has low costs， making it suitable for large-scale industrial production. BP-1 consists of six monosaccharides， contains α-glycosidic bonds and uronic acids， exhibits hypoglycemic activity， and provides a certain protective effect on the liver of alloxan-induced diabetic model zebrafish.

ObjectiveTo optimize the extraction method for polysaccharides from turnip（Brassica rapa）， and analyze and evaluate the primary structure of the isolated and purified turnip polysaccharide fraction（BP-1） and its hypoglycemic effects in diabetic zebrafish. MethodsTaking polysaccharide yield as the evaluation index， a semi-bionic extraction method was employed. Single-factor experiments and Box-Behnken response surface methodology were used to investigate three factors of solid-to-liquid ratio， extraction time and extraction temperature， in order to optimize the extraction process. BP-1 was isolated and purified using the Sevage method and DEAE-52 cellulose column chromatography. Structural characterization of the turnip polysaccharides was performed using ultraviolet-visible spectrophotometry（UV）， gas chromatography-mass spectrometry（GC-MS）， Congo red assay， and Fourier-transform infrared spectroscopy（FT-IR） to determine purity， monosaccharide composition， triple-helix structure， and functional groups. The microstructure of the polysaccharides was observed using scanning electron microscopy（SEM） and atomic force microscopy（AFM）. Zebrafish were divided into the blank group（adding E3 medium）， and BP-1-1， BP-1-10， BP-1-50， BP-1-200， BP-1-1 000 groups（adding BP-1 solutions at concentrations of 1， 10， 50， 200， 1 000 mg·L^-1， respectively）， and zebrafish embryos were subjected to a 96-hour exposure experiment. The maximum tolerated concentration of BP-1 in zebrafish was determined by evaluating its effects on phenotype， survival rate， malformation rate， and heart rate. Experimental animals were randomly divided into the blank group， model group， BP-1-10 group（10 mg·L^-1）， BP-1-50 group（50 mg·L^-1）， and BP-1-200 group（200 mg·L^-1）. The blank group was cultured in E3 medium， the model and treatment groups were induced to establish a diabetic model in 4 day-post-fertilization（dpf） zebrafish embryos using 10 g·L^-1 of glucose combined with 500 µmol·L^-1 of alloxan. The treatment groups received corresponding doses of BP-1 solution， while the blank and model groups received an equal volume of saline. Glucose and insulin（INS） levels were measured using enzyme-linked immunosorbent assay（ELISA） kits， the effects on the liver were observed by hematoxylin-eosin（HE） histopathological sections. The mRNA expression levels of glucagon（Glucagon）， insulin（Insa）， and phosphoenolpyruvate carboxykinase 1（PCK1） were detected with real-time fluorescence quantitative polymerase chain reaction（Real-time PCR）. ResultsThe optimized extraction conditions were determined as follows：solid-to-liquid ratio of 1∶40（g·mL^-1）， extraction time of 66 min， and extraction temperature of 79 ℃. Under these conditions， the yield of turnip polysaccharides was （10.34±0.96）%. UV analysis indicated that BP-1 contained no proteins or nucleic acids， GC-MS analysis revealed that BP-1 consisted of six monosaccharides（arabinose， rhamnose， ribose， mannose， galactose and glucose）. Congo red assay indicated that the molecular conformation did not exhibit a triple-helix structure， FT-IR analysis showed the presence of α-glycosidic bonds and uronic acids， SEM analysis revealed an irregular flaky structure with a flat and smooth surface， AFM analysis suggested that the aggregated structure might be formed by the entanglement of molecular chains and intramolecular hydrogen bonding. The maximum tolerated concentration of BP-1 in zebrafish over 96 h was determined to be 200 mg·L^-1. Pharmacodynamic results showed that， compared with the blank group， the model group exhibited significantly increased glucose levels and significantly decreased INS levels（P<0.01）. Compared with the model group， the BP-1-50 group significantly reduced glucose levels and increased INS levels（P<0.05）. Histopathological examination of liver tissue revealed that various doses of BP-1 had a certain reparative effect on damaged liver tissue. The liver tissue structure in the BP-1-200 group was nearly normal， with hepatocytes appearing plump. Real-time PCR results showed that， compared with the blank group， the model group exhibited significantly upregulated mRNA expressions of Glucagon and PCK1， and significantly downregulated mRNA expression of Insa（P<0.01）. Compared with the model group， the BP-1-50 and BP-1-200 groups showed significantly downregulated mRNA expressions of Glucagon and PCK1， and significantly upregulated mRNA expression of Insa（P<0.01）. ConclusionThe semi-bionic extraction method for turnip polysaccharides yields a high extraction rate， is simple to operate， has low costs， making it suitable for large-scale industrial production. BP-1 consists of six monosaccharides， contains α-glycosidic bonds and uronic acids， exhibits hypoglycemic activity， and provides a certain protective effect on the liver of alloxan-induced diabetic model zebrafish.

OBJECTIVE To investigate the effects of Brassica rapa polysaccharide （BRP） on the Toll-like receptor 4 （TLR4）/ myeloid differentiation factor 88 （MyD88）/nuclear factor-κB （NF-κB）， AMP-activated protein kinase （AMPK）/sterol regulatory element-binding protein-1c （SREBP-1c） pathways， intestinal microbiota and liver metabolism of mice with alcoholic liver injury， and preliminarily elucidate its mechanism for improving alcoholic liver injury. METHODS Seventy-two mice were randomly divided into blank group （normal saline）， model group （normal saline）， bifendate group （positive control， 300 mg/kg） and BRP low-， medium- and high-dose groups （75， 150 and 300 mg/kg）. They were given relevant medicine intragastrically， once a day， for consecutive 9 d. After the last administration， mice in all groups except the blank group were gavaged with white liquor to establish an alcoholic liver injury model. The levels of alanine aminotransferase and aspartate aminotransferase in serum， total cholesterol， triglycerides， low-density lipoprotein cholesterol， interleukin-6， interleukin-1β， tumor necrosis factor- α and lipopolysaccharide， as well as protein expressions of TLR4， MyD88， NF-κB p65， phosphorylated NF-κB p65 （p-NF-κB p65）， AMPK， phosphorylated AMPK （p-AMPK）， and SREBP-1c were all detected； pathological morphological changes of liver tissue and colon were observed. 16S rRNA was used to detect the changes of intestinal microbiota in mice， and metabolomics 2022B02058） technology was used to detect the changes of liver metabolites. RESULTS Compared with model group， the above biochemical indicators and the protein expressions of TLR4， MyD88， p-NF-κB p65， and SREBP-1c in liver tissues were all significantly decreased （P＜0.05 or P＜0.01）， while the protein expression of p-AMPK was significantly increased （P＜0.05 or P＜0.01）. Pathological damage to liver and colon tissues was significantly improved. Medium dose of BRP could increase the relative abundance of Akkermansia， norank_f_Muribaculaceae and Lachnospiraceae_NK4A136_group in the intestinal contents of mice to a certain extent， and decrease the relative abundance of Lactobacillus and Escherichia-Shigella. A total of 9 differential metabolites were identified by metabolomics， including homogentisic acid， myristyl lysophosphatidylcholine， which were involved in pathways such as tyrosine metabolism. CONCLUSIONS BRP can regulate the relative abundance of beneficial flora， reduce the relative abundance of harmful flora， improve the structure of intestinal colonies， reduce the entry of pro-inflammatory mediator lipopolysaccharides into liver tissue， affect metabolic pathways such as tyrosine metabolism and the expression of TLR4/MyD88/NF- κB and AMPK/SREBP-1c signaling pathways in the liver， and ultimately improve alcoholic liver injury.

ObjectiveTo analyze the pharmacodynamic activity of Bupleuri Radix processed with Trionyx sinensis blood in the treatment of Yin deficiency and study the spectrum-effect relationship of this medicine. MethodsHigh performance liquid chromatography was employed to establish the fingerprints of 15 batches of Bupleuri Radix processed with Trionyx sinensis blood， and the similarity was evaluated according to the SOP of Similarity Evaluation System of Chromatographic Fingerprint of TCM （version 2012）. A mouse model of Yin deficiency induced by thyroxine was established. The relationship between the active components and the effect on Yin deficiency was explored by grey correlation analysis and partial least squares method based on the changes in the serum levels of triiodothyronine （T3）， thyroxine （T4）， cyclic adenosine phosphate （cAMP）， and cyclic guanosine phosphate （cGMP）. The components screened out based on the spectrum-effect relationship were used for retrieval of the targets from the Traditional Chinese Medicine Systems Pharmacology and Analysis Database （TCMSP）， The Encyclopedia of Traditional Chinese Medicine （ETCM）， and Integrative Pharmacology-based Research Platform of Traditional Chinese Medicine （TCMIP）. Furthermore， the Online Mendelian Inheritance in Man （OMIM）， GeneCards， TTD， DisGeNET， and Drugbank were employed to establish the active component-target against Yin deficiency network of Bupleuri Radix processed with Trionyx sinensis blood. Gene Ontology （GO） and Kyoto Encyclopedia of Genes and Genomes （KEGG） pathway enrichment analyses were carried out for the core targets. Real-time PCR was conducted to verify the predicted key pathways and mechanisms. ResultsThe fingerprints of the 15 batches of Bupleuri Radix processed with Trionyx sinensis blood showed the similarities of 0.976-0.999 with the control fingerprint. Compared with the model group， the drug administration group showed elevated levels of T3 and T4 and lowered levels of cAMP， cGMP and cAMP/cGMP. The results of grey correlation analysis showed that active components in terms of the correlations followed the trend of saikosaponin B₁ > saikosaponin B₂ > saikosaponin C > saikosaponin D > saikosaponin A. The partial least squares analysis showed that saikosaponins A， D， B₁， and B₂ had higher VIP values. Network pharmacology predicted a total of 30 common targets， which were enriched in 276 GO terns and 115 KEGG pathways. The results of Real-time PCR showed that the model group had lower mRNA levels of Caspase-9， kinase insert domain receptor （KDR）， and mammalian target of rapamycin （mTOR） and higher mRNA level of mouse double minute 2 homolog （MDM2） than the blank group and the drug administration group. ConclusionBupleuri Radix processed with Trionyx sinensis blood has therapeutic effect on Yin deficiency syndrome， which provides a new idea for studying Bupleuri Radix processed with Trionyx sinensis blood.

Vesical vaginal fistula is the most common urogenital fistula. This article reportd a case treating by transurethral intravesical suturing combined with vaginal anterior wall suturing. The patient was followed up for 6 months after surgery, showing no symptoms, normal urine flow, and no recurrence of fistula. This surgical approach is simple to perform, minimally invasive, and has a quick recovery, which suggests the feasibility of this procedure. However, large sample studies are needed to verify its efficacy and safety.

Vesical vaginal fistula is the most common urogenital fistula. This article reportd a case treating by transurethral intravesical suturing combined with vaginal anterior wall suturing. The patient was followed up for 6 months after surgery, showing no symptoms, normal urine flow, and no recurrence of fistula. This surgical approach is simple to perform, minimally invasive, and has a quick recovery, which suggests the feasibility of this procedure. However, large sample studies are needed to verify its efficacy and safety.

【Objective】 To investigate the current status of incision sites to obtain intact specimens in laparoscopic nephrectomy by urologists in China, so as to provide reference for the standardized procedure. 【Methods】 During Jun.20, 2021 and Jul.4, 2021, more than 20 000 urologists in a WeChat group were surveyed with a questionnaire. The general data, incision sites and related complications were statistically analyzed. 【Results】 A total of 601 valid questionnaires were collected, covering urologists from 31 provinces, autonomous regions and municipalities. Surgical approaches: 68 urologists chose trans-abdominal approach, 432 chose posterior abdominal space approach, 101 chose both surgical approaches. Incision sites: 97 urologists chose lumbar transverse incision, 202 chose dorsal oblique incision of the waist, 119 chose ventral oblique incision, 93 chose the paramedian incision, 112 chose the lower abdominal oblique incision (Gibson), 11 chose the transverse lower abdominal incision (Pfannenstiel), 7 chose the median incision of the lower abdomen, 2 chose the median incision in the upper abdomen, 15 chose axillary midline direct incision; 399 chose to cut off the muscles, and 202 chose not to. Complications: 232 urologists reported pain after 2 weeks, 369 reported no pain; 325 reported numbness after 2 weeks, 276 reported no numbness; 66 reported incisional hernia, 535 reported no hernia. 【Conclusions】 Chinese urologists tend to choose retroperitoneoscopic nephrectomy and waist incision to obtain intact specimens. Transperitoneal laparoscopic nephrectomy has a variety of incisions for intact specimens. There is no standardized incision sites to obtain intact specimens.

Objective:To summarize the efficacy and safety of transurethral partial cystectomy combined with intravesical suturation in the treatment of bladder cancer.Methods:The clinical data of 28 patients with bladder cancer who were admitted in Zhengzhou People's Hospital from July 2020 to October 2021 were analyzed retrospectively. There were 22 males and 6 females, with the average age of (68.8±8.9) years old. Twenty cases were with single bladder tumor, and 8 cases were with multiple bladder tumors. The diameter of single bladder tumor measured by CT was less than 4 cm, with a median tumor diameter of 2.3(0.9, 2.8)cm. All 28 patients underwent transurethral partial cystectomy combined with intravesical suturation for the first time. Intraoperative intravesical instillation was conducted with 1 g gemcitabine. Bladder hot reperfusion treatment was performed with gemcitabine 2 g at 45℃ immediately after operation. The patients underwent reexamination with cystoscopy every 3 months after operation.Results:All the 28 patients successfully completed the operation. The average operation time was (128.3±16.3)min. No obturator nerve reflection was induced during operation. Muscularis tissue was found in all tumor pathological specimens after operation, among which 4 cases showed muscularis infiltration. Pathology of tumor basal and peripheral tissues showed tumor cell infiltration in 3 cases (1 case full-thickness infiltration, namely the incision margin was positive). The median continuous irrigation time of bladder after operation was 24.0(20.9, 25.0)h. The median time of indwelling catheter after operation was 6.5(6.0, 7.0)days, and one case developed severe bladder spasm. All patients were followed up, with a median follow-up of 7.5(4.0, 10.8) months.Postoperative recurrence occurred in 3 cases, all of which were ectopic recurrence in bladder.The 1-year recurrence rate was 10.7% (3/28).Conclusions:In the first surgical treatment of bladder cancer, transurethral partial cystectomy combined with intravesical suturation can provide accurate pathological staging, and low positive surgical margin rate. Immediate suturation of bladder wound during operation and immediate bladder perfusion chemotherapy can provide short bladder irrigation time, with few adverse reactions and low incidence of complications.

Central nervous system (CNS) injuries, including stroke, traumatic brain injury, and spinal cord injury, are essential causes of death and long-term disability and are difficult to cure, mainly due to the limited neuron regeneration and the glial scar formation. Herein, we apply extracellular vesicles (EVs) secreted by M2 microglia to improve the differentiation of neural stem cells (NSCs) at the injured site, and simultaneously modify them with the injured vascular targeting peptide (DA7R) and the stem cell recruiting factor (SDF-1) on their surface via copper-free click chemistry to recruit NSCs, inducing their neuronal differentiation, and serving as the nanocarriers at the injured site (Dual-EV). Results prove that the Dual-EV could target human umbilical vascular endothelial cells (HUVECs), recruit NSCs, and promote the neuronal differentiation of NSCs in vitro. Furthermore, 10 miRNAs are found to be upregulated in Dual-M2-EVs compared to Dual-M0-EVs via bioinformatic analysis, and further NSC differentiation experiment by flow cytometry reveals that among these miRNAs, miR30b-3p, miR-222-3p, miR-129-5p, and miR-155-5p may exert effect of inducing NSC to differentiate into neurons. In vivo experiments show that Dual-EV nanocarriers achieve improved accumulation in the ischemic area of stroke model mice, potentiate NSCs recruitment, and increase neurogenesis. This work provides new insights for the treatment of neuronal regeneration after CNS injuries as well as endogenous stem cells, and the click chemistry EV/peptide/chemokine and related nanocarriers for improving human health.

OBJECTIVE To conduct the pharmacoeconomic evaluation of empagliflozin in the treatment of heart failure with reduced ejection fraction （HFrEF），and to provide evidence-based reference for rational drug use and medical and healthy decision-making. METHODS A Markov model was used to perform a cost-effectiveness analysis of the regimen of empagliflozin in the treatment of HFrEF ，and to evaluate the cost and effectiveness of standard treatment plan plus empagliflozin （empagliflozin group）vs. standard treatment plan （standard treatment group ）. Clinical parameters were obtained from the EMPEROR-Reduced study；cost and utility data came from the published literatures. The cycle of the model was 1 month and the simulation time was 20 years. Single-factor sensitivity analysis and probability sensitivity analysis were performed to validate the results of cost-effectiveness analysis. RESULTS Compared with the standard treatment group ，each additional quality-adjusted life year in the empagliflozin group cost 37 995.94 yuan more ，which was less than China ’s 1 time GDP per capita in 2020（72 447 yuan）. The results of single factor sensitivity analysis showed that steady-state hospitalization rate of 2 groups was the most important factor affecting the incremental cost-effectiveness ratio . The results of probability sensitivity analysis showed that when the willingness-to-pay threshold （WTP）was 1 time GDP per capita in 2020（72 447 yuan），the probability of empagliflozin group with cost-effectiveness advantage was 58.8％；when the WTP was 3 times GDP per capita in 2020（217 341 yuan），the probability of empagliflozin group with cost-effectiveness advantage was 63.8％. CONCLUSIONS Compared with standard treatment plan alone，standard treatment plan plus empagliflozin is more cost-effective in the treatment of HFrEF. However ，the economic probability is not high.