ObjectiveTo investigate the therapeutic effects of Chaipo decoction on bronchial asthma in rats and its regulatory effects on the nucleotide-binding oligomerization domain-like receptor pyrin domain-containing protein 3 （NLRP3）/cysteinyl aspartate-specific protease-1 （Caspase-1）/Gasdermin D （GSDMD） pathway， aiming to elucidate its mechanism in ameliorating pyroptosis. MethodsSixty male Sprague-Dawley （SD） rats were randomly divided into six groups （n=10 per group）： normal control， asthma model， Chaipo decoction low-dose （5.0 g·kg^-1）， medium-dose （10.0 g·kg^-1）， high-dose （20.0 g·kg^-1）， and dexamethasone （1.0 mg·kg^-1）. The asthma model was established in all groups except the normal control group via ovalbumin （OVA） sensitization and challenge. Rats in the Chaipo decoction groups received intragastric administration of Chaipo decoction at the corresponding doses， while the dexamethasone group was treated with dexamethasone. The normal and model groups were administered equivalent volumes of saline. After 14 days of intervention， asthma symptom scores were assessed. Dynamic lung compliance （Cdyn）， lung resistance （RL）， and functional residual capacity （FRC） were measured using a small animal pulmonary function testing system. Lung tissue pathology was evaluated by hematoxylin-eosin （HE）， Masson's trichrome， and periodic acid-Schiff （PAS） staining. Levels of interleukin （IL）-6， IL-1β， and IL-18 in bronchoalveolar lavage fluid （BALF） were determined by enzyme-linked immunosorbent assay （ELISA）. Expression of NLRP3 and apoptosis-associated speck-like protein （ASC） in lung tissues was assessed by immunohistochemistry （IHC）. Protein levels of NLRP3， Caspase-1， GSDMD， and other pyroptosis-related proteins were measured by Western blot. ResultsCompared with the normal group， the model group exhibited significantly increased asthma symptom scores， inflammatory scores， collagen deposition， PAS scores， RL， FRC， levels of IL-6， IL-1β， and IL-18 in BALF， and expression levels of NLRP3， ASC， and other pyroptosis-related proteins in lung tissue （P0.01）， while Cdyn was significantly decreased （P0.01）. Compared with the model group， all doses of Chaipo decoction markedly improved asthma symptoms， with significantly reduced symptom scores （P0.05， P0.01）. Pulmonary function analysis showed that medium and high doses of Chaipo decoction significantly increased Cdyn （P0.05， P0.01） and decreased RL and FRC （P0.05， P0.01）. Histopathological evaluation indicated that high-dose Chaipo decoction significantly reduced inflammatory scores， collagen deposition， and goblet cell hyperplasia in lung tissue （P0.05， P0.01）. ELISA results showed that all doses of Chaipo decoction significantly decreased IL-6， IL-1β， and IL-18 levels in BALF （P0.05， P0.01）. IHC and Western blot analyses demonstrated that medium and high doses of Chaipo decoction markedly downregulated NLRP3， ASC， and other pyroptosis-related proteins in lung tissue （P0.05， P0.01）. ConclusionChaipo decoction effectively improves pulmonary function and pathological damage in asthmatic rats， potentially by inhibiting the NLRP3/Caspase-1/GSDMD pathway and reducing pyroptosis.

AIM:This study utilized CRISPR/Cas9 technology to create Retnlb floxp knock-in mice,followed by the application of the Cre-LoxP recombination system to generate intestinal epithelial-specific Retnlb gene knockout mice(Retnlb-CKO).This model was developed to investigate the pathogenic mechanisms of Retnlb in inflammatory bowel disease.METHODS:Female and male C57BL/6N mice,aged 8 weeks with the Retnlbflox/+genotype,were housed togeth-er for breeding.Offsprings were screened to identify those with the Retnlbflox/flox genotype.These mice were then crossed with Vil1-Cre transgenic mice,which express Cre recombinase specifically in intestinal epithelial cells,resulting in Retnlb-flox/+,Cre+mice.Subsequent crosses between Retnlbflox/+,Cre+mice and Retnlbflox/flox mice produced Retnlbflox/flox,Cre+mice(Retnlb-CKO).Six 8-week-old Retnlbflox/flox,Cre+mice and their littermate Retnlbflox/flox mice were selected for experiments.RT-qPCR and immunohistochemistry were used to assess Retnlb mRNA and protein levels in colonic epithelium.Phenotypic observa-tions included body length,weight,diet,and reproductive capability.Tissue-to-body weight ratios were calculated to ana-lyze growth and development.Intestinal barrier integrity and colonic expression of inflammatory factors were evaluated.RESULTS:The conditional gene knockout mouse model with specific deletion of Retnlb in intestinal epithelial cells was successfully established and validated through genetic identification,mRNA and protein analysis.Compared to Retnlbflox/flox mice,Retnlb-CKO mice exhibited no significant differences in body length,weight,diet,or reproductive capability.There were no differences in the ratios of heart,liver,spleen,lung,kidney,and colon weight to body weight,nor were there morphological differences in various tissues.However,the mRNA expression of tight junction proteins ZO-1,Occlu-din,and Claudin3 in colon tissues of Retnlb-CKO mice was significantly reduced(P<0.01).PAS staining and immunohis-tochemistry revealed a significant decrease in the number of goblet cells and lysozyme-positive cells in the colon tissues of Retnlb-CKO mice(P<0.01).HE staining showed no obvious pathological change in colon tissues of Retnlb-CKO mice.RT-qPCR further demonstrated a significant downregulation of pro-inflammatory factors NLRP3,interleukin-6(IL-6),IL-1β,and tumor necrosis factor-α(TNF-α)in colon tissues(P<0.01),along with significant downregulation of inflamma-tion signaling pathway proteins TLR4,MyD88,and NF-κB(P<0.01).CONCLUSION:A conditional colon epithelial cell Retnlb gene knockout mouse model was successfully constructed and validated.The absence of Retnlb in colon cells led to impaired intestinal barrier function,decreased mRNA expression of pro-inflammatory factors in colon tissue,and downregulation of mRNA expression of inflammatory pathway proteins TLR4,MyD88,and NF-κB.

This study was designed to explore the correlation between alterations in NLRP3 levels and Th17/Treg imbalance in asthmatic mice undergoing epithelial-mesenchymal transition(EMT).A murine model of asthma was established by intraperitoneal injection combined with nebulization of ovalbumin(OVA).Mice were randomly grouped into asthma model group and normal control group.The airway reactivity was detected with non-invasive lung function instrument.Hematoxylin and Eosin(HE)and Masson's trichrome staining were applied to evaluate the histopathological injury of lung tissue and the extent of lung fibrosis;RT-qPCR was applied to detect EMT-related biomarkers(Snail,E-Cadherin,N-Cadherin),the specific transcription factors of T cell subsets(RoRγt,Foxp3)and NLRP3 in lung tissue of mice;Western blot was used to detect the protein expression of E-cadherin,N-Cadherin and NLRP3 in lung tissue of mice.The Th17 and Treg cell populations in the spleen were enumerated via flow cytometry.Furthermore,the expression levels of NLRP3,IL-17 and IL-10 in bronchoalveolar lavage fluid(BALF)were analyzed by Giemsa staining.Compared with the control group,the asthma model group showed higher level of airway resistance,coupled with an obviously decrease in pulmonary ventilation compliance.Pathological alterations in lung tissue were evident,characterized by thickening of the airway epithelium,airway stenosis,infiltration of inflammatory cells,higher expression levels of N-Cadherin and NLRP3 proteins(P<0.05),lower expression level of E-Cadherin(P<0.001)and higher levels of marker genes(Snail and N-Cadherin)in lung tissue.Furthermore,model mice demonstrated higher level of NLRP3 in BALF(P<0.05),higher level of Th17 in spleen,and higher levels of retinoic acid orphan receptor(ROR)-γt mRNA(P<0.05)and Th17-related cytokines(IL-17)(P<0.01).Concurrently,model mice also showed an obviously decrease in the prevalence of Treg cells,Forkhead box Foxp3 mRNA(P<0.001),and Treg-related cytokine IL-10(P<0.05).The results of the Pearson correlation analysis indicated that the level of NLRP3 mRNA was positively correlated the ratio of RoR γt mRNA,but negatively correlated with Foxp3 mRNA in the lung tissue of asthmatic mice.Additionally,NLRP3 in BALF demonstrated a positive correlation with IL-17 and a negative correlation with IL-10.In conclusion,These findings suggest that NLRP3 may trigger bronchial EMT by exacerbating the immune imbalance of Th17/Treg cells.

Chronic refractory wound (CRW) is one of the most challengeable issues in clinic due to complex pathogenesis, long course of disease and poor prognosis. Experts need to conduct systematic summary for the diagnosis and treatment of CRW due to complex pathogenesis and poor prognosis, and standard guidelines for the diagnosis and treatment of CRW should be created. The Guideline forthe diagnosis and treatment of chronic refractory wounds in orthopedic trauma patients ( version 2023) was created by the expert group organized by the Chinese Association of Orthopedic Surgeons, Chinese Orthopedic Association, Chinese Society of Traumatology, and Trauma Orthopedics and Multiple Traumatology Group of Emergency Resuscitation Committee of Chinese Medical Doctor Association after the clinical problems were chosen based on demand-driven principles and principles of evidence-based medicine. The guideline systematically elaborated CRW from aspects of the epidemiology, diagnosis, treatment, postoperative management, complication prevention and comorbidity management, and rehabilitation and health education, and 9 recommendations were finally proposed to provide a reliable clinical reference for the diagnosis and treatment of CRW.

Objective: This study is designed to compare the changes of intestinal microflora and cytokine levels in patients with schizophrenia in different periods, analyze the correlation between intestinal microflora and disease symptoms, and explore the influence of intestinal microflora changes on the symptoms of patients with schizophrenia. Methods: 40 schizophrenic patients in different periods were included in the study, with their demographic data of age, body mass index(BMI), sex and course of disease collected. For each subject, serum was first collected for the levels of cytokines to be determined by an Meso Scale Discovery. The severity of schizophrenia was then assessed using negative and positive symptom scales. Finally macrofactor sequencing of intestinal flora was performed using MetaGeneMark. Results: Serum levels of interleukin-6(IL-6), interleukin-10(IL-10), interleukin-17(IL-17), interleukin-23(IL-23), and tumor necrosis factor-α(TNF-α) in the acute stage were higher than those in remission stage, and the results were statistically significant(P<0.05). Negative and positive symptom Scale(PANSS) positive factor scores were negatively correlated with TNF-α and IL-17(R=-0.312,P<0.05;R=-0.399,P<0.05); IL-23 was positively correlated with the negative factor score of the scale(R=-0.344,P<0.05). IL-6 was positively correlated with scale cognitive factor score(R=-0.339,P<0.05). IL-23 was positively correlated with the total score of the scale(R=-0.370,P<0.05). The microbial diversity detected in stool samples of patients with acute schizophrenia was lower than that of patients with remission, and certain difference was detected in the intestinal flora species composition between patients in the acute stage and in the remission stage. Conclusion The level of serum cytokines in the acute stage of schizophrenia is higher than that in the remission stage, and some cytokines levels are correlated with clinical symptoms.

Objective:To study the gene expression of nuclear factor erythroid-2-related factor 2 (Nrf2), glutathione-S-transferase (GST) and interleukin-1 β (IL-1β) in A549 cells exposed to hyperoxia and cell apoptosis after siRNA interference with Nrf2.Method:Normal A549 cell lines were assigned into normoxia+siRNA group, normoxia+control group, hyperoxia+siRNA group and hyperoxia+control group according to whether siRNA interference was used and the exposure environment (normoxia/hyperoxia). The hyperoxia environment contained 95%O 2 and 5%CO 2. The levels of mRNA expression of Nrf2, GST and IL-1β were detected using quantitative real-time polymerase chain reaction (qRT-PCR). Flow cytometry was used to examine cell apoptosis of the hyperoxia+control group and hyperoxia+siRNA group at different time points. Analysis of variance (ANOVA) was used to test the relative gene expression and apoptosis of A549 cells. Result:(1) Compared with the normoxia+control group, the expression of Nrf2 and GST in the hyperoxia+control group was significantly increased ( P<0.05), and the expression of IL-1β was significantly decreased ( P<0.05); the expression of Nrf2 and GST in the normoxia+siRNA group decreased significantly ( P<0.05), while the expression of IL-1β increased significantly ( P<0.05). (2) Compared with the normoxia+siRNA group, Nrf2 expression in the hyperoxia+siRNA group showed no significant changes ( P=0.230), GST expression increased slightly ( P=0.057), and IL-1β expression decreased slightly ( P=0.112). (3) Compared with the hyperoxia+control group, the expression of Nrf2 and GST in the hyperoxia+siRNA group decreased significantly ( P<0.05), and the expression of IL-1β increased significantly ( P=0.042). (4) Compared with the hyperoxia+control group, the apoptosis of A549 cells in the hyperoxia+siRNA group increased significantly at 24 h, 48 h and 72 h ( P<0.05). Conclusion:After interfering with Nrf2, siRNA may regulate the expression of GST and IL-1β, preventing oxidative stress, reducing inflammatory response and inhibiting apoptosis.

Objective:To explore the effects of operation timing on postoperative complications and operative duration in children undergoing non-emergency operation for femoral neck fracture.Methods:Fifty-nine children and adolescents with femoral neck fracture were retrospectively analyzed who had been admitted to Department of Pediatric Orthopedics, Zhengzhou Orthopedic Hospital Affiliated to Henan University from February, 2015 to September, 2019. They were 39 boys and 20 girls with a mean age of 11.7 (7.1, 13.7) years. By Delbet fracture classification, 7 cases were type Ⅰ, 27 cases type Ⅱ, 20 cases type Ⅲ, and 5 cases type Ⅳ. The patients were divided into 4 groups by the time from injury to operation (TFITO): 17 cases were assigned into group A with TFITO from 24 to 48 hours, 14 cases into group B with TFITO from 49 to 72 hours, 12 cases into group C with TFITO from 73 to 96 hours, and 16 cases into group D with TFITO>96 hours. The effects of TFITO on postoperative complications and operative duration were analyzed.Results:There were no significant differences between the 4 groups in their preoperative general data, showing they were comparable ( P>0.05). This cohort was followed up for 12 to 62 months (average, 20 months). The operation time was not included for this study in 6 cases whose associated injury had to be treated simultaneously. The median operation time for the other 53 patients was 80 (70, 105) min. The correlation coefficient between TFITO and operation time was 0.098 ( P=0.484). Postoperative complications occurred in 37.3% of the patients (22/59), including 14 cases of avascular necrosis of femoral head. For groups A, B, C and D, the incidences of complications were 47.1% (8/17), 50.0% (7/14), 25.0% (3/12) and 25.0% (4/16) while the incidences of avascular necrosis of femoral head 23.5% (4/17), 31.3% (5/16), 16.7% (2/12) and 18.8% (3/16), showing insignificant differences between the 4 groups in all the comparisons ( P>0.05). Conclusion:The time from injury to operation may not increase operative duration or postoperative complications such as avascular necrosis of femoral head in children undergoing non-emergency operation for femoral neck fracture.

Objective:To analyse the pathogenic bacteria distribution and clinical characteristics of late-onset sepsis (LOS) among premature infants with gestational age less than 34 weeks in Henan Province.Methods:The clinical data of 6 590 premature infants admitted to 17 medical institutions in Henan Province from January 2019 to December 2020 were retrospectively analyzed. The gestational age of infants was less than 34 weeks and was admitted to the neonatal ward within 7 days after birth. SPSS 19.0 statistical software was used for data analysis.Results:Among 6 590 premature infants LOS developed in 751 cases (11.40%), of whom the diagnosis was confirmed in 276 cases (36.75%) and 475 cases (63.25%) were diagnosed clinically. The fatality rate related to LOS was 13.58%. There were significant differences in the incidence of LOS and infection-related mortality among infants with different gestational ages and body weights ( χ2=388.894 and 13.572, χ2=472.282 and 9.257, P<0.05 or <0.01). Among 276 children with confirmed LOS, 286 strains of pathogenic bacteria were isolated. Gram-negative bacteria were most prevalent (178 strains), accounting for 62.24% of all infections, followed by fungi (58 strains, 20.28%). Klebsiella pneumoniae was most frequently detected Gram-negative bacteria (117 strains, 40.91%), among which 32.48% (38/117) was carbapenem-resistant Klebsiella pneumoniae. The proportion of diagnosed sepsis, the proportion of catheterization, and the infection-related mortality of infants with LOS in tertiary hospitals were all higher than those in secondary hospitals ( χ2=6.212, 5.313 and 4.435, all P<0.05). The proportion of exclusive breastfeeding in secondary hospitals was lower than that in tertiary hospitals ( χ2=19.216, P<0.05). The time of antibacterial drug use before infection in specialized hospitals was longer than that in general hospitals ( χ2=3.276, P<0.05). Conclusion:The incidence of LOS among preterm infants in Henan Province is high, which was mainly caused by Gram-negative bacteria. The clinical characteristics of LOS caused by different pathogens and in different health institutions are different, the prevention and control strategy should be developed accordingly to reduce the incidence LOS of preterm premature infants.

Objective To investigate the clinical result of treating Tillaux-Chaput fractures using the full ankle arthroscopy technique.Methods A retrospective analysis was made on 21 patients with Tillaux-Chaput fractures followed up after treatment by the full ankle arthroscopy technique from May 2013 to May 2015.There were 16 males and 5 females,with the age range of 6-55 years [(25.5 ± 12.8)years].Right ankle was involved in 12 patients and left ankle in 4 patients.Sixteen patients had single TillauxChaput fractures and 5 patients had combined proximal fibular fractures.Diagnosis of Tillaux-Chaput fractures was confirmed by X-rays in 18 patients an CT in 3 patients.Ankle arthroscopy was used through the anterolateral and anteriormedal approaches for closed reduction and internal fixation using one or two Herbert screws.Wound healing,bone union and visual analogue score (VAS) were detected postoperatively.Function assessment was performed using the American Orthopedic Foot and Ankle Society (AOFAS) ankle-hindfoot scoring system.Results All incisions healed by first intention,without evidence of nerve,vessel and tendon injuries.Period of follow-up was (15.3 ± 7.1) months (range,12-25 months).Fracture healed within 12 to 36 weeks [(22.6 ±4.6)weeks] after operation.VAS was (0.8 ± 0.3) points after operation,obviously lower than preoperative (8.3 ± 1.3) points (P ＜ 0.05).At the final follow-up,all patients regained normal ankle function and normal walking gait,without the presence of ankle pain and weight-bearing walking pain.AOFAS score was improved from preoperative (51.2 ± 12.5) points to postoperative (92.7 ± 16.5) points (P ＜ 0.05).Based on the AOFAS score,the results were excellent in 19 patients and good in 2 patients,with the excellent-good rate of 100％.Conclsion Full ankle arthroscopy by the anterolateral and anterionnedal approaches provides a precise and effective method for closed reduction and internal fixation of Tillaux-Chaput fractures and deserves clinical application.

Objective To investigate the clinical outcomes of using modified antegrade digital artery island flap for the treatment of the severely flexion contracture of the burned finger.Methods Between August,2013 to August,2015,21 patients (21 fingers) with severely flexion contracture of the burned finger were hospitalized for treatment.According to the Stren classification standard for the interphalangeal joint flexion contracture,all the patients were rated as type Ⅲ.The volar soft-tissue defect with exposed tendons,nerves,vessels or bone ranged from 1.0 cm × 2.0 cm to 2.5 cm × 4.0 cm after scar relaxation.The artery and the nerve defect were 1.5 to 4.5 cm and 2.0 to 4.2 cm,respectively.The wound were reconstructed with the modified antegrade digital artery island flap.The dorsal branches of the proper digital nerve of the flap were anastomosised with the proper digital nerve of the wound.The flap donor site was resurfaced with full-thickness skin grafting from inner aspect of the forearm.All the cases were called back for postoperative follow-up.Results All the reconstructed fingers and flaps survived completely without vascular problems.The donor skin graft survived and wound healed by first intention.All the patients were followed up with 11.5 months (range,6-22 months).The finger appearance was satisfactory.The texture and color of flaps in all cases were good.There was no pigmentation and contraction relapse.The contracted fingers received no cold intolerance.At the final examination,the average values of static 2-point discrimination were 5.2 mm (range,4.3-6.5 mm) of the flap.In the series,based on the Michigan Hand Outcome Questionnaire,18 patients were strongly satisfied with the injured finger appearance and 3 patients satisfied with the appearance.Conclusion The modified antegrade digital artery island flap,which is easy to raise with large flap size and can result with the good finger appearance and function,is an ideal technique for reconstruction of the severe flexion contracture of the burned finger.