OBJECTIVE To construct an insulin-resistant(IR)small intestinal organoid model of mice and study the protective effect of flavanomarein(FM)on the intestinal mucosal barrier in the model.METHODS ①Small intestinal organoid models of C57BL/6J and db/db of mice were constructed.The expressions of Ki-67,E-cadherin(E-cad),lysozyme(Lyz)and mucin-2(Muc-2)in small intestinal organ-oids were detected by 3D immunofluorescence.RT-qPCR was used to detect the expressions of fibro-nectin(Fn),glucagon-like peptide-1(GLP-1)and peotide YY(PYY)mRNA while Western blotting was used to detect the expressions of Fn,GLP-1 and PYY protein.The Lyz secretion level was detected by ELISA.② Small intestinal organoids were divided into five groups:C57BL/6J mice 'small intestinal organ-oids as the normal control group,db/db mice' intestinal organoids as the IR model group,db/db mice small intestinal organoids with flavanomarein 25,50 and 100 μmol·L-1 intervention for 48 h as IR model+ FM groups.RT-qPCR was used to detect the expression of Lyz mRNA while Western blotting was used to detect the expression of Lyz protein.RESULTS ① On the 6th day of small intestinal organoid culture,a ring structure with a clear luminal structure was formed and an IR mouse small intestinal organoid model was established.3D Immunofluorescence detection showed that the established small intestinal organoids all expressed Ki-67,E-cad,Lyz and MUC-2.Compared with the normal control group,the expres-sion of Fn mRNA in the IR model group was significantly increased(P<0.05)while the expressions of GLP-1 and PYY mRNA were significantly decreased(P<0.05).Compared with the normal control group,the expression of Fn protein in the IR model group was significantly decreased(P<0.05)while the expressions of GLP-1 and PYY protein were significantly increased(P<0.05).ELISA results showed that compared with the normal control group,the secretion levels of Lyz in the IR model group were signifi-cantly decreased(P<0.01).② RT-qPCR results showed that compared with the normal control group,the expression of Lyz mRNA in the IR model group was significantly decreased(P<0.01).Compared with the IR model group,the expression of Lyz mRNA in the IR model+FM 50 and 100 μmol·L-1 groups was significantly increased(P<0.05,P<0.01).Western blotting results showed that compared with the normal control group,the expression of Lyz protein in the IR model group was significantly decreased(P<0.01).Compared with the IR model group,the expression of Lyz protein in the IR model+FM 50 and 100 μmol·L-1 groups was significantly increased(P<0.05,P<0.01).CONCLUSION The constructed IR mouse small intestinal organoid model provides a more complete in vitro research model for exploring the pathophysiological mechanism by which drug interventions help repair the intestinal mucosal barrier.FM may maintain the intestinal mucosal barrier by reversing the decrease in Lyz expression levels in IR mice,thereby improving IR.

Objective:To explore the clinical effect of using great toe fibular flaps of both feet on reconstruction of pulp defects of two neighbouring digits.Methods:A total of 14 digit pulp defects in 7 cases were repaired in Zhoukou Huaihai Hospital using great toe fibular flaps of both feet from August 2020 to January 2023. Of the 7 cases, there were 4 males and 3 females, with an average of 28 years old, ranging from 19 to 45 years old. Meanwhile, there were 4 cases in left hand and 3 cases in right hand. There were 3 cases of digit pulp defects in index and middle fingers, 2 in middle and ring fingers, and 2 in thumb and index fingers. The area of soft tissue defect in 1.2 cm×1.5 cm-3.0 cm×2.5 cm, and flap was 1.5 cm ×1.8 cm-3.2 cm×2.8 cm. Furthermore, 1 case underwent emergency surgery and 5 were repaired in elective surgery. The donor site of the flap was closed directly, and an intermediate-thickness skin graft was prepared from the medial plantar area for transfer in the case of high suture tension at the wound edge. After surgery, patients received postoperative by outpatient clinic and WeChat to observe the appearance, sensation, functional recovery and flap contracture of digits, as well as the movement of the great toes of both feet.Results:After the surgery, all flaps in the 7 cases survived smoothly and the donor sites healed. All patients entered scheduled follow-ups postoperatively for 6 months to 2 years, with an average of 9 months. The flap showed an aesthetic appearance and excellent sensation, with a TPD of 3-6 mm, and satisfactory digit function. The donor site of the great toe fibular flap left linear scars only, without abnormality in range of motion and gait in walking. In addition, there were 5 in excellent and 2 in good according to the Evaluation Trial Standards of Upper Limb Partial Functional of Hand Surgery of Chinese Medical Association.Conclusion:Application of great toe fibular flaps of both feet is an ideal option for the simultaneous repair of pulp defects of two neighbouring digits, which can achieve good reconstructive results.

Objective To explore the influence of extracellular matrix protein ABI-interactor 3-binding protein(ABI3BP)on vesicular stomatitis virus(VSV)genome replication and innate immune signaling pathway. Methods The small interfering RNA(siRNA)was transfected to knock down ABI3BP gene in human skin fibroblast BJ-5ta cells.VSV-green fluorescent protein(VSV-GFP)-infected cell model was established.The morphological changes and F-actin stress fiber formation were detected on ABI3BP knockdown cells by phalloidin immunofluorescence staining.The mRNA level of virus replication was detected by RT-qPCR in BJ-5ta cells after VSV-GFP infection;western blotting was performed to detect the changes in interferon regulatory factor 3(IRF3)andTANK-binding kinase 1(TBK1)phosphorylation levels. Results The VSV-GFP-infected BJ-5ta cell model was successfully established.Efficient knockdown of ABI3BP in BJ-5ta cells was achieved.Phalloidin immunofluorescence staining revealed structural rearrangement of intracellular F-actin after ABI3BP gene knockdown.Compared with the control group,the gene copy number of VSV-GFP in ABI3BP knockdown cells increased by 2.2-3.5 times(P＜0.01)and 2.2-4.0 times(P＜0.01)respectively when infected with VSV of multiplicity of infection 0.1 and 1.The expression of viral protein significantly increased in ABI3BP knockdown cells after virus infection.The activation of type-Ⅰ interferon pathway,as determined by phosphorylated IRF3 and phosphorylated TBK1,was significantly decreased in ABI3BP knockdown cells after VSV-GFP infection. Conclusions Extracellular matrix protein ABI3BP plays an important role in maintaining the formation and rearrangement of actin structure.ABI3BP gene deletion promotes RNA virus replication,and ABI3BP is an important molecule that maintains the integrity of type Ⅰ interferon pathway.

Objective To explore the value of quantitative chest CT in early diagnosis of chronic obstructive pulmonary disease(COPD).Methods The clinical data of 138 cases of COPD high-risk patients in Shanghai community and COPD high-risk respondents in outpatient clinic of Zhongshan Hospital,Fudan University from September 20,2013 to May 20,2014 were retrospectively analyzed.All high-risk participants underwent pulmonary function and chest CT examination at baseline and 1 year later.Chest CT images were imported into quantitative CT analysis software to collect quantitative CT data.These participants were divided into COPD group(n=40)and non-COPD group(n=98)based on their lung functions after 1 year.The differences in baseline lung function and quantitative CT measurements between the two groups were compared.Binary logistic regression was used to analyze the predictors of COPD in high-risk individuals after 1 year of follow-up,and the efficacy of the logistic regression model was evaluated by ROC curve.Results There were no significant differences in gender,body mass index(BMI),the percentage value of forced expiratory volume in 1 second predicted(FEV1%pred),airway wall area ratio(WA%),total airway count(TAC),and airway wall thickness(WT)between the two groups at baseline.Compared to the non-COPD group,the square root of the tracheal wall area at 10 mm from the inner circumference of the airway(Pi10),(3.43[3.30,3.54]vs 3.23[3.15,3.36],P＜0.001),and the percentage area of low attenuation regions below ﹣950 HU(LAA%﹣950),(2.06[0.32,6.29]vs 0.57[0.25,1.89],P=0.015)were significantly higher in the COPD group.The mean lung density(MLD)in the COPD group was lower than that in the non-COPD group([﹣799.89±35.62]vs[﹣783.60±43.52],P=0.038).Binary logistic regression analysis indicated that age and Pi10 were risk factors for COPD(P＜0.05),with an area under the ROC curve of 0.791(95%CI 0.714-0.868).Conclusions In the COPD high-risk population with normal lung function,patients with elevated Pi10 and LAA%﹣950 have a higher incidence of COPD one year later,suggesting that quantitative chest CT measurements such as Pi10 and LAA%﹣950 can assist clinicians in identifying early-stage COPD.

Objective To investigate the impact of corona virus disease 2019(COVID-19)vaccination on incidence of COVID-19 pneumonia and pulmonary function in early convalescence phase.Methods A total of 1 178 COVID-19 patients from Zhongshan hospital,Fudan University were enrolled from 7 Jan 2023 to 7 Feb 2023 and divided into two groups according to the incidence of COVID-19 pneumonia.All the patients were asked to finish the questionnaire on COVID-19 related-symptoms,and other clinical parameters in our study.Vaccination situation,Pulmonary functions including ventilation,diffusion function and fractional exhaled nitricoxide(FeNO)were collected.T-tests and Chi-squared tests were used to analyze the protective effect of vaccination on pulmonary function and occurrence of COVID-19 pneumonia between groups.Univariate analysis and multivariate analysis were performed to predict the significant prognostic factors for incidence of COVID-19 pneumonia.Results Lung function including ventilation and diffusion function was significantly declined in unvaccinated patients than vaccinated patients(P<0.05).Those unvaccinated patients and patients with chronic diseases were more likely to have COVID-19 pneumonia and severe related symptoms.Vaccinated with two or three doses,rather than one dose of COVID-19 vaccines could protect the patients from COVID-19 pneumonia(P<0.05).Those with chronic diseases were inclined to have severe symptoms,while vaccination could alleviate the symptoms of those patients and patients with no history.Conclusion COVID-19 infection can impair the lung function of patients in early convalescence phase.COVID-19 vaccination could protect the patients from COVID-19 pneumonia,ameliorate the related symptoms and attenuate lung function decline.

Aim To investigate the protective poten-tial of paeoniflorin 6-oxy-benzenesulfonate(CP-25)in preserving knee cartilage integrity in osteoarthritis mice through inhibition of GRK2 activity.Methods The posttraumatic osteoarthritis model was established fol-lowing DMM surgery.The experiment consisted of a sham operation group,a model group,a CP-25 admin-istration group,and a paroxetine positive control group.Intragastric administration commenced after the surgery.After 12 weeks of administration,the animals were euthanized.Micro-CT imaging was used to ob-serve the knee cartilage degeneration and abnormal bone remodeling,and the joint histopathology of mice was observed by staining with ferrubens solid green.Immunohistochemistry and immunofluorescence were used to detect the expression level of related molecules in cartilage tissue.Furthermore,Western blot was em-ployed to determine GRK2 and EP4 membrane protein expression levels as well as total protein levels of GRK2 and MMP13 following CP-25 treatment.Results Compared with the sham operation group,the articular cartilage in the model group was significantly degrad-ed,with the cartilage surface calcifying and osteo-phytes forming.CP-25 could significantly reduce the number of osteophytes and the thickness of the sub-chondral plate of articular cartilage,promote the regen-eration of the cartilage matrix,reduce the expression of cartilage matrix degradation protein,and have a signifi-cant protective effect on knee cartilage.Immunohisto-chemical and immunofluorescence results showed that compared with the model group,CP-25 treatment sig-nificantly decreased the expressions of GRK2,AD-AMTS5 and MMP13 in knee tissue and increased the expressions of Col Ⅱ and Aggrecan in knee tissue.The results of in vitro experiments showed that CP-25 ad-ministration could significantly reduce the expression levels of GRK2 membrane protein and total protein,in-crease the level of EP4 membrane protein,and de-crease the level of MMP13.Conclusions The ad-ministration of CP-25 can significantly promote the re-generation of articular cartilage matrix in OA mice,re-duce the degradation of cartilage matrix,and exhibit therapeutic effects on OA.The mechanism behind this is related to the inhibition of GRK2-mediated cartilage matrix metabolism.

Heme oxygenase-1(HO-1)is an inducible heme oxygenase and a catalytic enzyme for heme decomposition reactions,which can catalyze the heme decomposition into CO,biliverdin and Fe2+.HO-1 and its metabolites have anti-inflammatory,antioxidant and anti-apoptotic effects in human body,and play an important role in neurodegenerative diseases such as Alzheimer's disease,Parkinson's disease,amyotrophic lateral sclerosis,and Huntington's disease.This article will review the production,distribution,and gene structure of HO-1,the biological characteristics of its metabolites,and the role and mechanism of HO-1 in neurodegenerative diseases,in order to provide a theoretical basis for the clinical application of HO-1.

Objective:To explore the clinical efficacy of free superficial circumflex iliac artery perforator flap (SCIAPF) in reconstruction of soft tissue defects of hand and foot in children.Methods:From August 2021 to August 2023, free SCIAPFs were used to reconstruct soft tissue defects of hands and feet in 8 children at the Department of Hand and Foot Surgery, Zhoukou Huaihai Hospital. There were 6 boys and 2 girls aged between 2 and 7 years old. The sites of soft tissue defect were: 1 of metacarpophalangeal joint in the functional area of left thumb, 2 of dorsal right hand, 1 of dorsal left foot, 2 of medial malleolus of right foot, and 2 of right forefoot, and all were accompanied with tendon and bone exposure. The size of soft tissue defects ranged from 2.5 cm×3.0 cm to 6.0 cm×3.5 cm. After emergency debridement, the children received flap transfers, the flaps sized from 3.0 cm×3.5 cm to 7.0 cm×4.0 cm. Donor sites were closed by running intradermal suture. All children were included in the scheduled postoperative follow-up by visit of outpatient clinic, WeChat interviews and home visits to observe the appearance, colour, texture and sensation of the flaps, the functional recovery of the affected hands and feet, as well as the healing of donor sites.Results:After surgery, all 8 flaps survived beside 1 had a partial necrosis at the edge and healed with a scab after dressing change. Flap donor sites healed in one stage. Scheduled postoperative follow-up lasted from 7 to 22 months, with an average of 10 months. The flaps presented a slightly bloated appearance, soft texture with a restored protective sensation and satisfactory functional recovery of hands and feet. Linear scars were left at the donor sites. According to the Evaluation Trial Standards of Upper Limb Partial Functional of Hand Surgery of Chinese Medical Association, 1 child was of excellent and 2 children of good in hand function of the 3 children with hand injury. The Maryland Foot Score revealed 3 children were of excellent and 2 of good in foot function of the 5 children with foot injury.Conclusion:Free SCIAPF is an ideal option for reconstruction of soft tissue defects of hand and foot in children, with concealed donor site, convenient flap harvesting, fewer complications and satisfactory reconstructive effect.

AIM: To explore the key genes and molecular markers involved in the retinoblastoma development through bioinformatics.METHODS: The mRNA microarray datasets from the Gene Expression Omnibus(GEO)database were obtained, and the differentially expressed gene(DEG)between retinoblastoma cell lines and normal retinal pigment epithelial(RPE)cell lines were analyzed through gene ontology(GO)and KEGG enrichment analysis. To screen key genes, establish protein-protein interaction(PPI)network, and use receiver operating characteristic(ROC)curve to assess clinical diagnostic efficacy. The RNA expressions of key genes in retinoblastoma cell lines and normal RPE cell lines were compared by qRT-PCR.RESULTS: A total of 121 DEGs were obtained from the retinoblastoma dataset of GSE97508 and GSE110811. KEGG pathway analysis showed that DEG were enriched in phototransduction, cell cycle, and p53 signaling pathways. A total of 9 key genes, including MCM6, DTL, UBE2T, TOP2A, NUSAP1, CENPK, RRM2, RLBP1, and RHO, were obtained from the intersection of PPI network analysis and the top 30 DEG from each dataset. The differentially expressed 9 key genes were verified in GSE24673. ROC analysis showed that the area under the curve(AUC)for UBE2T, RRM2, and RHO was ≥80%, and there was a statistical significance(P>0.05). The mRNA level of UBE2T and RRM2 in retinoblastoma was significantly higher than APRE-19 cell line, while the mRNA level of RHO was significantly lower than that of ARPE-19 cell line.CONCLUSION: UBE2T, RRM2, and RHO may be served as potential molecular markers and potential therapeutic targets for retinoblastoma.

AIM: To evaluate the lacrimal gland parameters and their correlation with clinical examination in patients with thyroid-associated ophthalmopathy(TAO)using orbital magnetic resonance imaging(MRI).METHODS: A total of 38 patients(76 eyes)with TAO were selected as case group, and 26 patients(52 eyes)who matched the gender and age with case group and volunteered to accept examination were selected as normal control group. Patients in case group were categorized into active TAO group and inactive TAO group according to the modified clinical activity score(CAS). The exophthalmos was evaluated on T1WI after obtaining the MRI images, the longest lacrimal gland length, width, and the biggest area in axial and coronal images were evaluated on T2WI, and the maximum T2 value and mean T2 value of the lacrimal gland were recorded.RESULTS: There were no significant differences in age, gender and exophthalmos between active TAO and inactive TAO(P>0.05). The area of lacrimal gland was higher in active TAO than that in inactive TAO, and was higher in inactive TAO than that in control group in coronal and axial section(all P<0.01). The length of lacrimal gland in coronal and axial section was higher in the active TAO than that in the inactive TAO and the control group(all P<0.05). The width of lacrimal gland in coronal and axial section was higher in active TAO and inactive TAO than that in the control group(all P<0.05). The maximum T2 value in the active TAO was higher than that in the inactive TAO and control group, and the inactive TAO was higher than that in the control group(all P<0.05). The average T2 value in the active TAO was higher than that in the inactive TAO and control group(all P<0.05). CAS was positively correlated with lacrimal gland area in axial, coronal section and maximum T2 value(all P<0.01).CONCLUSION: The lacrimal gland is significantly enlarged in patients with TAO, especially in active TAO. The lacrimal gland area in axial, coronal section and maximum T2 value could be potentially utilized as valuable radiographic biomarkers for the activity of TAO.