1.Extraction,Separation and Hypoglycemic Activity Analysis of Polysaccharides from Brassica rapa
Mengyu HOU ; Ruina XU ; Qingsong LI ; Shaoxuan LI ; Xinying MA ; Yaohui YE
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(10):219-228
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.
2.Extraction,Separation and Hypoglycemic Activity Analysis of Polysaccharides from Brassica rapa
Mengyu HOU ; Ruina XU ; Qingsong LI ; Shaoxuan LI ; Xinying MA ; Yaohui YE
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(10):219-228
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.
3.Spectrum-effect Relationship of Bupleuri Radix Processed with Trionyx sinensis Blood for Yin Deficiency Based on Saponins
Mengyu HOU ; Xia ZHAO ; Zhiyu GUO ; Ting LIU ; Yuexing MA ; Yaohui YE
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(3):147-155
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.
4.Study on the mechanism of Brassica rapa polysaccharide in improving alcoholic liver injury of mice based on intestinal microbiota and metabolomics
Xinying MA ; Ruina XU ; Shaoxuan LI ; Ruiyin YE ; Yuexing MA ; Yaohui YE
China Pharmacy 2025;36(16):2005-2011
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.
5.Primary exploration of stage I anastomosis and T-tube fistulation in laparoscopic local resection of duodenal tumors
Lijie LUO ; Tao WANG ; Xinrui YE ; Xianzhe WANG ; Zhuoxuan ZHANG ; Zijing ZHANG ; Yaohui PENG ; Yan CHEN ; Haiping ZENG ; Haipeng TANG ; Jiantao LIN ; Weiqiang ZOU ; Wei WANG
Chinese Journal of Gastrointestinal Surgery 2025;28(2):198-202
Objective:To discuss the feasibility and safety of stage I anastomosis and T-tube fistulation in laparoscopic local resection of duodenal tumors.Methods:A descriptive case series study was used to retrospectively analyze the clinical diagnosis and treatment data of 14 patients with duodenal tumors who successfully underwent laparoscopic local resection of duodenal tumors + phase I anastomosis + T-tube ostomy in the Guangdong Provincial Hospital of Chinese Medicine and the First Affiliated Hospital of Guangzhou University of Chinese Medicine from October 2021 to March 2024. The resection and reconstruction steps of laparoscopic local resection of duodenal tumor + phase I anastomosis + T-tube ostomy are as follows: (1) after the safe margin is clear, the duodenal tumor is completely removed in full thickness, and the specimen bag is taken out and sent to frozen section to determine the nature of the tumor and the negative margin; (2) Perforate the anterior duodenal wall below the tumor plane, place a 16# T tube, and fix it with laparoscopic purse string suture. The abdominal wall is led out through the duodenum, and the duodenal T tube fistulation is performed; (3) The duodenum was continuously sutured in a full-thickness transverse shape, and the seromuscular layer was strengthened to form a phase I anastomosis. The nutritional improvement of patients after operation was mainly observed, and the intraoperative situation and postoperative complications were recorded.Results:No conversion to laparotomy, postoperative emergency reoperation, intraoperative and postoperative complications occurred in 14 patients with duodenal tumors who completed laparoscopic local resection of duodenal tumors + phase I anastomosis + T-tube ostomy. The operation time was (225.43 ± 56.54) min, and the intraoperative blood loss was (72.14 ± 74.65) ml. The patient recovered well after operation, and no severe postoperative abdominal bleeding occurred. Postoperative gastrointestinal angiography showed that the anastomotic stoma was unobstructed, and there were no stenosis, anastomotic leakage and other related complications. There was no significant difference in serum albumin [(37.09 ± 3.53) g/L vs. (37.52 ± 4) g/L] and hemoglobin [(100.79 ± 31.93) g/L vs. (103.07 ± 19.6) g/L] between before and 1 week after operation ( P > 0.05). Conclusion:Laparoscopic local resection of duodenal tumor + phase I anastomosis + T-tube fistulation can be used as one of the safe and feasible improved methods for local resection of duodenal tumor to effectively reduce the occurrence of related complications.
6.Primary exploration of stage I anastomosis and T-tube fistulation in laparoscopic local resection of duodenal tumors
Lijie LUO ; Tao WANG ; Xinrui YE ; Xianzhe WANG ; Zhuoxuan ZHANG ; Zijing ZHANG ; Yaohui PENG ; Yan CHEN ; Haiping ZENG ; Haipeng TANG ; Jiantao LIN ; Weiqiang ZOU ; Wei WANG
Chinese Journal of Gastrointestinal Surgery 2025;28(2):198-202
Objective:To discuss the feasibility and safety of stage I anastomosis and T-tube fistulation in laparoscopic local resection of duodenal tumors.Methods:A descriptive case series study was used to retrospectively analyze the clinical diagnosis and treatment data of 14 patients with duodenal tumors who successfully underwent laparoscopic local resection of duodenal tumors + phase I anastomosis + T-tube ostomy in the Guangdong Provincial Hospital of Chinese Medicine and the First Affiliated Hospital of Guangzhou University of Chinese Medicine from October 2021 to March 2024. The resection and reconstruction steps of laparoscopic local resection of duodenal tumor + phase I anastomosis + T-tube ostomy are as follows: (1) after the safe margin is clear, the duodenal tumor is completely removed in full thickness, and the specimen bag is taken out and sent to frozen section to determine the nature of the tumor and the negative margin; (2) Perforate the anterior duodenal wall below the tumor plane, place a 16# T tube, and fix it with laparoscopic purse string suture. The abdominal wall is led out through the duodenum, and the duodenal T tube fistulation is performed; (3) The duodenum was continuously sutured in a full-thickness transverse shape, and the seromuscular layer was strengthened to form a phase I anastomosis. The nutritional improvement of patients after operation was mainly observed, and the intraoperative situation and postoperative complications were recorded.Results:No conversion to laparotomy, postoperative emergency reoperation, intraoperative and postoperative complications occurred in 14 patients with duodenal tumors who completed laparoscopic local resection of duodenal tumors + phase I anastomosis + T-tube ostomy. The operation time was (225.43 ± 56.54) min, and the intraoperative blood loss was (72.14 ± 74.65) ml. The patient recovered well after operation, and no severe postoperative abdominal bleeding occurred. Postoperative gastrointestinal angiography showed that the anastomotic stoma was unobstructed, and there were no stenosis, anastomotic leakage and other related complications. There was no significant difference in serum albumin [(37.09 ± 3.53) g/L vs. (37.52 ± 4) g/L] and hemoglobin [(100.79 ± 31.93) g/L vs. (103.07 ± 19.6) g/L] between before and 1 week after operation ( P > 0.05). Conclusion:Laparoscopic local resection of duodenal tumor + phase I anastomosis + T-tube fistulation can be used as one of the safe and feasible improved methods for local resection of duodenal tumor to effectively reduce the occurrence of related complications.
7.Protective effect of Humanin on rotenone-induced dopamine neuron toxicity
Yaohui SHAN ; Qifu ZHANG ; Jin CHENG ; Feng YE ; Xi ZHANG ; Wenpei YU ; Xiaogang WANG ; Yuanpeng ZHAO ; Guorong DAN ; Mingliang CHEN ; Yan SAI
Journal of Army Medical University 2024;46(7):670-677
Objective To investigate the mechanism and protective effect of Humanin(HN)on rotenone(Rot)-induced toxic damage for dopamine neurons.Methods The Rot-poisened PC12 cell model was constructed,and the control group,the Rot poisening group,the HN pretreated Rot poisening group,and the HN treatment group were set up.ELISA was used to detect the content of HN inside and outside of Rot-infected cells,CCK-8 assay was used to detect cell viability,and ATP detection kit was used to detect the intracellular ATP content.Dichloro-dihydro-fluorescein diacetate(DCFH-DA)assay was used to detect the level of reactive oxygen species(ROS)in cells.Western blotting was performed to detect the expression level of mitochondrial autophagy regulatory proteins Pink1,Parkin,p62,LC3,mitochondrial biogenesis regulatory protein PGC1α,division/fusion regulatory proteins OPA1,MFN2,DRP1,p-DRP1 and antioxidant stress regulatory proteins Keap1 and Nrf2.HBAD-mcherry-EGFP-LC3 adenovirus transfected cells was used to observed the number of autophagosomes and autophagolysosomes.Results The results showed that the intracellular concentration of HN in PC12 in the Rot poisening group was significantly higher than that in the control group(P<0.05);Compared with the control group,the Rot poisening group had significantly decreased activity of PC12 cells,decreased ATP content and increased production of ROS.After the poisen of Rot in PC12 cells,the expression of Pink1 and p-Parkin,the ratio of LC3Ⅱ/LC3Ⅰ and the expression of p-DRP1 in mitochondrial fusion protein was increased,while the expression of p62,the expression of mitochondrial biogenesis protein PGC1 α,mitochondrial fusion proteins MFN2 and OPA1,and antioxidant stress proteins Keap1 and Nrf2 were decreased(all P<0.05).The number of autophagosomes and autophagolysosomes in PC12 cells in the Rot poisening group was higher than that in the control group(P<0.05),and HN pretreatment(20 μmol/L)could significantly improve the changes mentioned above caused by Rot poisening(P<0.05).Conclusion HN ameliorates Rot-induced toxic damage for dopamine neurons by inhibiting mitophagy and mitochondrial division and promoting mitochondrial biogenesis and fusion,and anti-oxidative stress.
8.Evaluation of A Liver Yin Deficiency Mouse Model Based on Untargeted Metabolomics
Xuening HUANG ; Xia ZHAO ; Mengyu HOU ; Zhiyu GUO ; Yaohui YE
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(5):118-125
ObjectiveBased on ultra performance liquid chromatography-quadrupole-time-of-flight tandem mass spectrometry(UPLC-Q-TOF-MS), to evaluate the establishment of a mouse model of liver Yin deficiency by thyroid tablet suspension combined with 10% carbon tetrachloride(CCl4) from the perspective of non-targeted metabolomics, in order to lay the foundation for the establishment of a traditional Chinese medicine(TCM) syndrome model. MethodA total of 24 mice were randomly divided into blank group and model group. The model group was given thyroid tablet suspension(0.003 2 g·kg-1) by gavage for 14 consecutive days, and 10% CCl4(5 mL·kg-1) was intraperitoneally injected once a week to establish a liver Yin deficiency model, while the blank group was injected with an equal amount of olive oil intraperitoneally and gavaged with an equal amount of distilled water, and was fed with normal feed. After the modeling was completed, 6 mice in each group were randomly selected, the levels of alanine aminotransferase(ALT), aspartate aminotransferase(AST), cyclic adenosine monophosphate(cAMP), cyclic guanosine monophosphate(cGMP), interleukin(IL)-6, IL-10, tumor necrosis factor-α(TNF-α)were measured in the mice serum, and malondialdehyde(MDA), superoxide dismutase(SOD), total protein(TP), hydroxyproline(HYP) and other indicators were measured in the mice liver. Liver tissue sections were taken for hematoxylin-eosin(HE) staining and observing pathological changes. The remaining 6 mice in each group were subjected to UPLC-Q-TOF-MS combined with principal component analysis(PCA) and orthogonal partial least squares-discriminant analysis(OPLS-DA) were used to screen differential metabolites in the liver Yin deficiency mouse model, Kyoto Encyclopedia of Genes and Genomes(KEGG) database was used to analyze the corresponding metabolic pathways of differential metabolites. ResultCompared with the blank group, mice in the model group showed liver Yin deficiency manifestations such as reduced body weight, fatigue and sleepiness, disheveled and lusterless hair, irritability. The levels of ALT, cAMP/cGMP, IL-6, AST, MDA, cAMP, TNF-α significantly increased(P<0.05, P<0.01), while the levels of SOD, IL-10 and cGMP significantly decreased(P<0.05, P<0.01), and the changes of HYP and TP were not statistically significant. Hepatic steatosis and distortion of the radial arrangement of the liver plate cells were seen in the section images of the model group, endogenous substances were clearly separated, and 252 differential metabolites were identified in the serum samples, which were mainly involved in the metabolic pathways of purine metabolism, steroid hormone biosynthesis and pyrimidine metabolism. A total of 229 differential metabolites were identified in the liver samples, mainly involving nucleotide metabolism, purine metabolism, steroid hormone biosynthesis, pyrimidine metabolism, antifolate resistance, insulin resistance, primary bile acid biosynthesis, prostate cancer, sulfur relay system, arachidonic acid metabolism and other metabolic pathways. ConclusionThe successful establishment of liver Yin deficiency model in mice by CCl4 combined with thyroid hormone is evaluated through the investigation of serum and liver metabolomics, combined with biochemical indicators, which provides a biological basis and experimental foundation for the Yin deficiency syndrome model of TCM.
9.Application of anterior esophageal wall full layer fixation and gastric tube guidance in total laparoscopic overlap method for intracorporeal esophagojejunostomy
Yan CHEN ; Xinrui YE ; Lijie LUO ; Zijing ZHANG ; Wenjun XIONG ; Haigang YANG ; Yaohui PENG ; Zeyu LIN ; Zhuoxuan ZHANG ; Wei WANG
Chinese Journal of Gastrointestinal Surgery 2024;27(10):1074-1079
Objective:To explore the application of anterior esophageal wall full layer fixation and gastric tube guidance in total laparoscopic overlap method for intracorporeal esophagojejunostomy.Methods:Overlap esophagojejunostomy with anterior esophageal wall full layer fixation and gastric tube guidance is suitable for patients with advanced gastric cancer (clinical stage: cT1b~4aN0~3M0) and esophageal invasion <3 cm, who underwent radical total gastrectomy+ overlap esophagojejunostomy. The main operation procedure was performed as follows: A titanium clip was used for fixation of the full anterior wall of esophagus before overlap esophagojejunostomy, and the side‐to‐side esophagojejunostomy was performed with the linear stapler under the guidance of gastric tube. Then the titanium clip was removed after confirming that the correct cavity was entered. Finally, the common outlet was closed by two barbed sutures. A descriptive case series study was conducted. The clinical data of patients who underwent laparoscopic radical gastrectomy and overlap esophagojejunostomy with anterior esophageal wall full layer fixation and gastric tube guidance in Guangdong Provincial Hospital of Chinese medicine and the First Affiliated Hospital of Guangzhou University of Chinese medicine from May 2021 to June 2023 were retrospectively analyzed.Results:A total of 42 patients were collected, and all of them were successfully completed laparoscopic total radical gastrectomy without conversion to laparotomy or perioperative death. The esophagojejunostomy time, operative time, intraoperative blood loss was 17(5‐25) minutes, (258.8±38.0) minutes and 50(20‐200) ml, respectively. The incidence of esophageal false lumen was 0%, and there were no intraoperative complications. The time of gastric tube removal, initial fluid diet intake and the duration of postoperative hospital were 2(1‐5) , 4(1‐8) and 8(4‐21) days, respectively. There were no postoperative anastomotic hemorrhage, anastomotic stenosis and other related complications. One patient (2.38%) developed a Clavien‐Dindo IIIb complication, which was abdominal hemorrhage after operation. The second surgical exploration confirmed that the patient was bleeding due to gastroduodenal artery rupture. After intraoperative suture hemostasis, fluid expansion, blood transfusion and other treatments, the patient was discharged on the 15th day after the operation. Three patients (7.14%) developed Clavien‐Dindo grade II complications, including anastomotic leakage, chylous leakage and pulmonary infection, and were discharged after conservative treatment such as anti‐infection and prolonged retention of drainage tube.Conclusions:Laparoscopic overlap method for intracorporeal esophagojejunostomy with anterior esophageal wall fixation and gastric tube guidance can shorten the time of esophagojejunostomy and prevent the occurrence of false lumen, and do not increase anastomose‐related complications.
10.Application of anterior esophageal wall full layer fixation and gastric tube guidance in total laparoscopic overlap method for intracorporeal esophagojejunostomy
Yan CHEN ; Xinrui YE ; Lijie LUO ; Zijing ZHANG ; Wenjun XIONG ; Haigang YANG ; Yaohui PENG ; Zeyu LIN ; Zhuoxuan ZHANG ; Wei WANG
Chinese Journal of Gastrointestinal Surgery 2024;27(10):1074-1079
Objective:To explore the application of anterior esophageal wall full layer fixation and gastric tube guidance in total laparoscopic overlap method for intracorporeal esophagojejunostomy.Methods:Overlap esophagojejunostomy with anterior esophageal wall full layer fixation and gastric tube guidance is suitable for patients with advanced gastric cancer (clinical stage: cT1b~4aN0~3M0) and esophageal invasion <3 cm, who underwent radical total gastrectomy+ overlap esophagojejunostomy. The main operation procedure was performed as follows: A titanium clip was used for fixation of the full anterior wall of esophagus before overlap esophagojejunostomy, and the side‐to‐side esophagojejunostomy was performed with the linear stapler under the guidance of gastric tube. Then the titanium clip was removed after confirming that the correct cavity was entered. Finally, the common outlet was closed by two barbed sutures. A descriptive case series study was conducted. The clinical data of patients who underwent laparoscopic radical gastrectomy and overlap esophagojejunostomy with anterior esophageal wall full layer fixation and gastric tube guidance in Guangdong Provincial Hospital of Chinese medicine and the First Affiliated Hospital of Guangzhou University of Chinese medicine from May 2021 to June 2023 were retrospectively analyzed.Results:A total of 42 patients were collected, and all of them were successfully completed laparoscopic total radical gastrectomy without conversion to laparotomy or perioperative death. The esophagojejunostomy time, operative time, intraoperative blood loss was 17(5‐25) minutes, (258.8±38.0) minutes and 50(20‐200) ml, respectively. The incidence of esophageal false lumen was 0%, and there were no intraoperative complications. The time of gastric tube removal, initial fluid diet intake and the duration of postoperative hospital were 2(1‐5) , 4(1‐8) and 8(4‐21) days, respectively. There were no postoperative anastomotic hemorrhage, anastomotic stenosis and other related complications. One patient (2.38%) developed a Clavien‐Dindo IIIb complication, which was abdominal hemorrhage after operation. The second surgical exploration confirmed that the patient was bleeding due to gastroduodenal artery rupture. After intraoperative suture hemostasis, fluid expansion, blood transfusion and other treatments, the patient was discharged on the 15th day after the operation. Three patients (7.14%) developed Clavien‐Dindo grade II complications, including anastomotic leakage, chylous leakage and pulmonary infection, and were discharged after conservative treatment such as anti‐infection and prolonged retention of drainage tube.Conclusions:Laparoscopic overlap method for intracorporeal esophagojejunostomy with anterior esophageal wall fixation and gastric tube guidance can shorten the time of esophagojejunostomy and prevent the occurrence of false lumen, and do not increase anastomose‐related complications.

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