BACKGROUND:Flap transplantation technique is a commonly used surgical procedure for the treatment of severe tissue defects,but postoperative flap necrosis is easily triggered by ischemia-reperfusion injury.Therefore,it is still an important research topic to improve the survival rate of transplanted flaps. OBJECTIVE:To review the pathogenesis and latest treatment progress of flap ischemia-reperfusion injury. METHODS:CNKI,WanFang Database and PubMed database were searched for relevant literature published from 2014 to 2024.The search terms used were"flap,ischemia-reperfusion injury,inflammatory response,oxidative stress,Ca2+overload,apoptosis,mesenchymal stem cells,platelet-rich plasma,signaling pathways,shock wave,pretreatment"in Chinese and English.After elimination of irrelevant literature,poor quality and obsolete literature,77 documents were finally included for review. RESULTS AND CONCLUSION:Flap ischemia/reperfusion injury may be related to pathological factors such as inflammatory response,oxidative stress response,Ca2+overload,and apoptosis,which can cause apoptosis of vascular endothelial cells,vascular damage and microcirculation disorders in the flap,and eventually lead to flap necrosis.Studies have found that mesenchymal stem cell transplantation,platelet-rich plasma,signaling pathway modulators,shock waves,and pretreatment can alleviate flap ischemia/reperfusion injuries from different aspects and to varying degrees,and reduce the necrosis rate and necrosis area of the grafted flap.Although there are many therapeutic methods for skin flap ischemia/reperfusion injury,a unified and effective therapeutic method has not yet been developed in the clinic,and the advantages and disadvantages of various therapeutic methods have not yet been compared.Most of the studies remain in the stage of animal experiments,rarely involving clinical observations.Therefore,a lot of research is required in the future to gradually move from animal experiments to the clinic in order to better serve the clinic.

OBJECTIVE To optimize the water extraction technology for Kaixin granules. METHODS UPLC-MS/MS method was established for the simultaneous determination of ginsenoside Rg1， ginsenoside Re， ginsenoside Rb1， tenuifolin， polygalaxanthone Ⅲ and 3， 6′-disinapoyl-sucrose. An orthogonal test was designed with extraction times， extraction duration， and the volume of added water as factors. Using the contents of the aforementioned six indicator components and the extract yield as evaluation indexes， analytic hierarchy process-entropy weight method was employed to determine the combined weights of each indicator. Subsequently， process optimization and validation were conducted by integrating grey relational analysis （GRA） and back propagation （BP） neural network. RESULTS The water extraction technology optimized by the orthogonal test and GRA was 10- fold water for the first decoction and 8-fold water for the subsequent two， extracting 3 times，extracting for 1 h each time； the average comprehensive score of the validation experiment was 91.10 （RSD＝0.31%， n＝3）. The water extraction technology optimized by BP neural network was extracting 3 times with 10-fold water added each time， extracting for 1.5 h each time； the average comprehensive score of the validation experiment was 95.89 （RSD＝0.73%， n＝3）. Considering practical production requirements， the optimal water extraction technology was extraction performed three times， with 10-fold water for the first decoction and 8-fold water for the subsequent two extractions， with an extraction time of 1 h each. CONCLUSIONS The optimized water extraction technology for Kaixin granules is stable and feasible.

The enterotoxigenic Escherichia coli (ETEC) infection is a major factor restricting the development of animal husbandry. However, the abuse of antibiotics will lead to the antibiotic residues and emergence of antibiotic-resistant bacteria. The existing vaccines face challenges in stimulating intestinal immunity, demonstrating limited prevention effects. Therefore, it is indispensable to develop a new vaccine that is safe and suitable as a feed additive to activate intestinal immunity. This study constructed yeast-cell microcapsules (YCM) carrying the DNA vaccine against ETEC by genetic engineering. Furthermore, animal experiments were carried out to explore the regulatory effects of feeding YCM on the intestinal immune system and intestinal microbiota. Saccharomyces cerevisiae was selected as the oral delivery vehicle (microcapsules) of the DNA vaccine. The codon-optimized nucleic acid sequence of K88, the main antigen of mammal-derived ETEC, was synthesized, and the yeast shuttle vector containing the corresponding DNA vaccine expression cassette was constructed by DNA recombination. The recombinant strain of YCM was prepared by transforming JMY1. Additionally, the characteristics of the YCM strain and its feasibility as an oral vaccine were comprehensively evaluated by the fluorescence reporter assay, gastrointestinal fluid tolerance assay, intestinal epithelial cell adhesion assay, intestinal retention assessment, antiserum detection, and intestinal microbiota detection. The experimental results showed that the DNA vaccine expression cassette was expressed in mammals, and the recombinant strain of YCM could tolerate up to 8 hours of gastrointestinal fluid digestion and had good adhesion to intestinal epithelial cells. The results of mouse feeding experiments indicated that the recombinant strain of YCM could stay in the intestinal tract for at least two weeks, and the DNA vaccine expression cassette carried by YCM entered the intestinal immune system and triggered an immune response to induce the production of specific antibodies. Moreover, feeding YCM recombinant bacteria also improved the abundance of gut microbiota in mice, demonstrating a positive effect in regulating intestinal flora. In summary, we prepared the recombinant strain of YCM carrying the DNA vaccine against ETEC and comprehensively evaluated its characteristics and feasibility as an oral vaccine. Feeding the recombinant YCM could induce specific immune responses and regulate intestinal microbiota. The findings provide a reference for the immunoprevention of ETEC-related animal diseases.
Animals ; Enterotoxigenic Escherichia coli/genetics* ; Saccharomyces cerevisiae/metabolism* ; Vaccines, DNA/genetics* ; Mice ; Escherichia coli Infections/immunology* ; Escherichia coli Vaccines/genetics* ; Capsules ; Mice, Inbred BALB C ; Female

BACKGROUND:In recent years,it has been found that some traditional Chinese medicine monomers can alleviate oxidative stress and apoptosis of the skin flap,promote vascular regeneration of the skin flap and prevent skin flap necrosis by activating autophagy. OBJECTIVE:To review the research progress of traditional Chinese medicine monomer regulating autophagy in preventing flap necrosis. METHODS:The Chinese and English key words were"traditional Chinese medicine(TCM),autophagy,skin flaps".The first author searched the relevant articles published in CNKI and PubMed databases from January 2010 to October 2022.A total of 196 articles were retrieved in the preliminary screening and then screened according to the inclusion and exclusion criteria.The quality assessment was conducted by reading the literature titles and abstracts.Finally,55 articles were summarized. RESULTS AND CONCLUSION:(1)The regulation of autophagy is mediated by AMPK/mTOR,PI3K/AKT and other signaling pathways.Activation of autophagy can alleviate the oxidative stress and apoptosis of the flap,promote the regeneration of blood vessels in the flap,and prevent flap necrosis.(2)Terpenoids(Betulinic acid,Andrographolide,Notoginseng Triterpenes,Catalpa),phenolic compounds(Resveratrol,Curcumin,Gastrodin),phenolic acids(Salvianolic acid B)and steroid compounds(Pseudoginsenoside F11)in traditional Chinese medicine monomers can alleviate oxidative stress and apoptosis of skin flap by regulating related signaling pathways to activate autophagy,promote skin flap angiogenesis and promote skin flap survival.(3)Studying the research progress of traditional Chinese medicine monomer to prevent flap necrosis by regulating autophagy can provide a reference and theoretical basis for traditional Chinese medicine to prevent flap necrosis and promote flap healing in the clinic.

BACKGROUND:Mesenchymal stem cells have great potential in the treatment of ischemia-reperfusion injury of skin flaps.However,their defects and the decline of their role in the treatment of ischemia-reperfusion injury of skin flaps restrict their wide application. OBJECTIVE:To review the strategies for improving the treatment of ischemia-reperfusion injury of skin flaps with mesenchymal stem cells,and provide a reference for its further theoretical research and clinical application. METHODS:Relevant documents included in CNKI,WanFang and PubMed were searched.The Chinese and English search terms were"mesenchymal stem cell,ischemia-reperfusion adjustment of skin flap,mesenchymal stem cells,stem cells,skin flap,ischemia-reperfusion injury,pretreatment,gene modification,biomaterial packaging,joint application".The relevant documents since 2007 were retrieved,and the documents with little relationship between the research content and the article theme,poor quality and outdated content were eliminated through reading the article,and finally 75 documents were included for summary. RESULTS AND CONCLUSION:(1)Mesenchymal stem cells can inhibit inflammatory reactions,resist oxidative stress and induce angiogenesis,which has great potential in the treatment of skin flap ischemia-reperfusion injury.(2)Although mesenchymal stem cells have shown great potential in the treatment of skin flap ischemia-reperfusion injury,their shortcomings in treatment have limited their widespread clinical application.Through pre-treatment(cytokines,hypoxia,drugs,and other pre-treatment mesenchymal stem cells),gene-modified mesenchymal stem cells,biomaterial encapsulation of mesenchymal stem cells,as well as the combined use of mesenchymal stem cells and other drugs or therapeutic methods,can not only overcome the shortcomings of mesenchymal stem cells in treatment,but also improve their therapeutic effectiveness in skin flap ischemia-reperfusion injury.(3)Therefore,further improving the effectiveness of mesenchymal stem cells in treating skin flap ischemia-reperfusion injury and exploring its therapeutic potential are of great significance for the research of mesenchymal stem cells and the treatment of skin flap ischemia-reperfusion injury.

BACKGROUND:Mesenchymal stem cells are used in flap ischemia-reperfusion injury due to their antioxidant and inflammatory inhibition,and angiogenesis induction. OBJECTIVE:To review the mechanism and latest treatment progress of mesenchymal stem cells in the treatment of flap ischemia-reperfusion injury,and to provide a basis for further theoretical research and clinical rational application. METHODS:We searched the relevant articles indexed in CNKI,WanFang and PubMed databases.Chinese and English search terms were"mesenchymal stem cells;flap ischemia reperfusion injury;conditioned medium;exosomes;oxidative stress;inflammatory reactions;angiogenesis".Relevant literature since 2010 was searched,and 74 articles were finally included after excluding the literature that had little to do with the topic of the article,poor quality and outdated content. RESULTS AND CONCLUSION:(1)Mesenchymal stem cells play significant roles in antioxidation,inhibition of inflammation and induction of angiogenesis and have great potential in the treatment of flap ischemia-reperfusion injury.(2)However,the defects of mesenchymal stem cells themselves and the decline of therapeutic effect in recent years have put the development and application of mesenchymal stem cells into a bottleneck period,and the research on the plasticity of mesenchymal stem cells conditioned medium and its exosomes and mesenchymal stem cells came into being,and the therapeutic effect was significantly better than the use of mesenchymal stem cells alone.(3)Therefore,a more comprehensive understanding of the mechanism of action and the latest treatment progress of mesenchymal stem cells in the treatment of flap ischemia-reperfusion injury is of great significance for the research of mesenchymal stem cells and the treatment of flap ischemia-reperfusion injury.

Objective:To predict and analyze the T/B combined epitope of EM10 protein in Echinococcus multilocularis, and identify the expressed products of the biosynthetic EM10 multi epitopes. Methods:The gene-related information of EM10 protein was obtained through NCBI GenBank public database. Bioinformatics technique was used to predict and analyze the T/B binding epitopes of EM10 protein. The prokaryotic expession recombinant plasmid pET30a-EM10 (epitope) was synthesized, and transformed into host bacteria Ecoli. BL21 (DE3). The expression of EM10 recombinant multi-epitope protein was identified by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blotting after induced expression by isopropyl thiogalactopyranoside (IPTG). Results:The total length of EM10 gene was 1 759 bp (GenBank registration number: U05573), and its protein amino acid sequence (GenBank registration number: AAA50580.1) was 559 amino acids. By using Phyre software for homology modeling, the tertiary structure of EM10 protein was obtained, and the T/B combined epitope of EM10 protein was successfully predicted, the dominant epitope was located at 46 - 61, 133 - 183, 239 - 255 and 442 - 475 amino acid sites. The (GGGGS)n linker sequence was used to connect the epitopes to form an EM10 recombinant multi-epitope protein with a total of 206 amino acid. The size of the DNA fragment was 618 bp and the relative molecular weight of the protein was 22.66 × 10 3. The prokaryotic expession recombinant plasmid was validated by enzyme digestion, the results showed that the plasmid size was between 5 000 and 6 000 bp, which was consistent with the length of the constructed plasmid (5 854 bp). SDS-PAGE showed that the target protein was expressed in the supernatant induced by IPTG at 37 ℃ and the effect was the best. The relative molecular weight of the protein was 22.66 × 10 3 by Western blotting, which was consistent with the constructed plasmid. Conclusions:The combined epitope of EM10 T/B is successfully designed and predicted using bioinformatics technology. A prokaryotic expression recombinant plasmid is constructed, the expression of EM10 recombinant multi-epitope protein is verified through experiments, providing an experimental basis for the construction of an EM10 dominant epitope diagnostic kit.

Objective To evaluate the efficacy of transanal opening of intersphincteric space(TROPIS)for the treatment of complex anal fistula.Methods 41 patients with complex anal fistula were randomly divided into two groups by random envelope method:21 patients in the treatment group were treated with TROPIS;20 patients in the control group were treated with traditional low incision and high thread drawing surgery.The curative effect,postoperative pain score,wound healing time,treatment failure rate,anal sphincter function and postoperative complications were compared between the two groups.Results The visual analoguescale scores(VAS)of anal pain in the treatment group and the control group 8 hours,24 hours,3 days,7 days and the first defecation after operation were(1.81±1.12)vs.(5.00±1.49),(1.10±1.14)vs.(4.35±1.42),(0.86±1.01)vs.(4.35±1.27),(0.81±1.08)vs(4.25±1.41),(3.05±1.56)vs(6.70±1.17),respectively,with significant differences(P＜0.01).The VAS of anal pain in the treatment group and the control group 14 days after operation were(0.67±1.07)vs(0.80±1.11),respectively,with no significant difference(P＞0.05).1 case in the treatment group(4.76％)and 1 case in the control group(5.00％)had wound infection after surgery,there was no significant difference in the rate of wound infection between the two groups(P＞0.05).2 cases(9.52％)in the treatment group and 3 cases(15.00％)in the control group underwent postoperative catheterization,there was no significant difference in urinary retention between the two groups(P＞0.05).In the treatment group,16 cases were cured,3 cases were markedly effective,and the total effective rate was 90.48％,while in the control group,16 cases were cured,2 cases were markedly effective,and the total effective rate was 90.00％,there was no significant difference between the two groups(P＞0.05).The wound healing time of the treatment group and the control group were(37.31± 3.42)days vs(48.13±4.08)days,respectively,with a statistically significant difference(P＜0.01).12 months after operation,2 patients in the treatment group and 1 patient in the control group lost the follow-up.12 months after operation,the ST Marks anal incontinence scores in the treatment group and the control group were(0.53±1.07)and(1.74±2.77),respectively,with a statistically significant difference(P＜0.05).There were 6 patients(31.58％)in the treatment group and 4 patients(21.05％)in the control group who failed in treatment,there was no significant difference in the rate of treatment failure between the two groups(P＞0.05).Conclusion The TROPIS is reliable in treating anal fistula,and has the advantages of less pain,quick recovery and less damage to anal function.

【Objective】 To investigate the preoperative epidemiological factors affecting the surgical outcomes of transurethral resection of the prostate (TURP) in patients with benign prostatic hyperplasia (BPH) and to evaluate the clinical predictive value. 【Methods】 Patients undergoing TURP at our hospital during Dec.2018 and Sep.2021 whose pathological examination suggestive of BPH were involved. Before operation, the clinical data, International Prostate Symptom Score (IPSS), quality of life (QoL) score, overactive bladder (OAB) score, prostate-specific antigen (PSA), and urological ultrasound results were collected. After operation, all patients were followed up with telephone call or face-to-face interview, and the IPSS, QoL score and OAB score were examined. Patients were divided into effective and ineffective groups according to the efficacy rubrics. The epidemiological factors affecting the efficacy of TURP were identified with univariate analysis, the independent influencing factors were screened with binary logistic regression, and the diagnostic value of each independent influencing factor was evaluated using receiver operating characteristic (ROC) curve. 【Results】 OAB score (OR=0.749, 95%CI: 0.627-0.895, P=0.001), IPSS-V/S (OR=4.919, 95%CI: 1.617-14.963, P=0.005), history of urinary retention (OR=7.513, 95%CI: 2.289-24.656, P=0.001), and history of urinary incontinence (OR=2.656, 95%CI: 1.015-6.950, P=0.047) were independent influencing factors for poor postoperative outcomes. ROC curve revealed that the area under the ROC curve (AUC) showed that AUC of OAB score, IPSS-V/S, history of urinary retention, and history of urinary incontinence were 0.784 (95%CI: 0.718-0.841), 0.686 (95%CI: 0.614-0.751), 0.713 (95%CI: 0.643-0.777), and 0.723 (95%CI: 0.654-0.786), respectively. ROC curve of the regression model showed that the AUC was 0.888 (95%CI: 0.834-0.930), and the sensitivity and specificity were 93.53% and 67.35%, respectively. 【Conclusion】 Preoperative OAB score, IPSS-V/S, history of urinary retention, and history of urinary incontinence were independent epidemiological factors of poor outcomes after TURP in BPH patients. The diagnostic efficacy ranked from the highest to the lowest as regression model >OAB score>history of urinary incontinence >history of urinary retention>IPSS-V/S.

Flap surgery is an important method to repair large open wounds in trauma and plastic surgery. However， flap necrosis is a common issue. Good blood circulation that provides nutrition is a necessary condition for the survival of the flap. Promoting revascularization， establishing blood circulation， and restoring blood perfusion as soon as possible after flap transplantation is crucial for flap survival. However， revascularization/blood reperfusion can cause flap ischemia-reperfusion injury （FIRI） after flap repair. If FIRI is not treated correctly and timely， it can cause flap necrosis and graft failure eventually. Previous studies have shown that the signaling pathways related to cell proliferation and apoptosis， such as the Notch signaling pathway， mitogen-activated protein kinase （MAPK） signaling pathway， and phosphatidylinositol 3-kinase/protein kinase B （PI3K/Akt） signaling pathway， and inflammation-related pathways， such as nuclear factor E₂-related factor 2 （Nrf2） signaling pathway， and nuclear factor-κB （NF-κB） signaling pathway can promote angiogenesis， improve local blood supply， inhibit the production and expression of inflammatory mediators， reduce oxidative stress and apoptosis， relieve ischemia-reperfusion injury （IRI）， and promote rapid healing after flap repair. By consulting a large number of modern medical research literature， this study reveals that both Chinese medicine monomers and Chinese medicine compounds can promote revascularization， restore blood perfusion， and accelerate healing after flap repair by regulating the expression of key protein molecules in the above-mentioned signaling pathways. This study summarized the research status in China and abroad， aiming to provide references for the in-depth study and clinical application of Chinese medicine to promote the healing of skin flaps after repair.