ObjectiveTo investigate the mechanism by which Feixin decoction treats hypoxic pulmonary hypertension （HPH） by regulating the peroxisome proliferator-activated receptor gamma （PPARγ）/nuclear factor-kappa B （NF-κB）/NOD-like receptor pyrin domain containing 3 （NLRP3） signaling pathway. MethodsForty-eight male SD rats were randomly allocated into normal， hypoxia， and low-， medium- and high-dose （5.85， 11.7， 23.4 g·kg^-1， respectively） Feixin decoction groups， with 8 rats in each group. Except the normal group， the remaining five groups were placed in a hypoxia chamber with an oxygen concentration of （10.0±0.5）% for 8 h per day， 28 days， and administrated with corresponding drugs during the modeling process. After 4 weeks of treatment， echocardiographic parameters ［pulmonary artery acceleration time （PAT）， pulmonary artery ejection time （PET）， right ventricular anterior wall thickness （RVAWd）， and tricuspid annular plane systolic excursion （TAPSE）］ were measured for each group. The right ventricular systolic pressure （RVSP） was measured by the right heart catheterization method， and the right ventricular hypertrophy index （RVHI） was calculated by weighing the heart. The pathological changes in pulmonary arterioles were observed by hematoxylin-eosin staining. The co-localization of α-smooth muscle actin （α-SMA） with NLRP3， N-terminal gasdermin D （N-GSDMD）， and cysteinyl aspartate-specific proteinase-1 （Caspase-1） in pulmonary arteries was detected by immunofluorescence. The protein levels of PPARγ， NF-κB， NLRP3， apoptosis-associated speck-like protein containing a CARD （ASC）， N-GSDMD， interleukin-1β （IL-1β）， interleukin-18（IL-18）， and cleaved Caspase-1 in the lung tissue was determined by Western blot. The ultrastructural changes in pulmonary artery smooth muscle cells （PASMCs） were observed by transmission electron microscopy. ResultsCompared with the normal group， the hypoxia group showed increased RVSP and RVHI （P<0.01）， decreased right heart function （P<0.01）， increased pulmonary vascular remodeling （P<0.01）， increased co-localization of α-SMA with NLRP3， N-GSDMD， and Caspase-1 in pulmonary arterioles （P<0.01）， up-regulated protein levels of NF-κB， NLRP3， ASC， N-GSDMD， IL-1β， IL-18， and cleaved Caspase-1 in the lung tissue （P<0.05， P<0.01）， a down-regulated protein level of PPARγ （P<0.05， P<0.01）， and pyroptosis in PASMCs. Compared with the hypoxia group， Feixin decoction reduced RVSP and RVHI， improved the right heart function and ameliorated pulmonary vascular remodeling （P<0.05， P<0.01）， decreased the co-localization of α-SMA with NLRP3， N-GSDMD， and Caspase-1 （P<0.05， P<0.01）， down-regulated the protein levels of NF-κB， NLRP3， ASC， N-GSDMD， IL-1β， IL-18， and cleaved Caspase-1 in the lung tissue （P<0.05， P<0.01）， up-regulated the protein level of PPARγ （P<0.05， P<0.01）， and alleviated pyroptosis in PASMCs. ConclusionFeixin decoction can ameliorate pulmonary vascular remodeling and right heart dysfunction in chronically induced HPH rats by regulating pyroptosis in PASMCs through the PPARγ/NF-κB/NLRP3 pathway.

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.

Objective To investigate the effect and mechanism of Feixin Decoction(Astragali Radix,Pericae Semen,Carthami Flos,Descurainiae Semen Lepidii Semen,Paeoniae Radix Rubra,etc.)on monocrotaline-induced pulmonary arterial hypertension(PAH)rats based on peroxisome proliferator-activated receptor-γ/nuclear factor-κB(PPAR-γ/NF-κB)signaling pathway.Methods Forty-eight male SD rats were randomly divided into normal group,model group,Sildenafil group(0.025 g·kg-1)and low-,medium-and high-dose of Feixin Decoction groups(11.7,23.4,46.8 g·kg-1).PAH rat model was established by single intraperitoneal injection of monocrotaline solution(60 mg·kg-1).After 1 hour of modeling,the rats were given intragastric administration once a day for 28 days.Hemodynamic and echocardiographic parameters including right ventricular systolic pressure(RVSP),mean pulmonary artery pressure(mPAP),right ventricular hypertrophy index(RVHI),pulmonary artery acceleration time(PAAT),pulmonary artery ejection time(PET),tricuspid annular plane systolic excursion(TAPSE),right ventricular internal diameter(RVIDd)and right ventricular anterior wall thickness(RVAWT)were measured in each group.The pathological changes of pulmonary arterioles were observed by HE staining.The expression level of α-smooth muscle actin(α-SMA)in rat pulmonary artery was detected by immunofluorescence.The levels of plasma interleukin-1β(IL-1β),IL-6 and tumor necrosis factor-α(TNF-α)were detected by ELISA.The expression levels of PPAR-γ/NF-κB signaling pathway-related proteins were detected by immunohistochemistry and Western Blot.Results Compared with the normal group,the RVSP,mPAP,RVHI,RVIDd and RVAWT of the model group were significantly increased(P＜0.01).PAAT,PAAT/PET and TAPSE were significantly decreased(P＜0.01).The wall of pulmonary arterioles was significantly thickened,and the percentage of wall thickness of pulmonary arterioles to vascular diameter and the percentage of vascular wall area to total cross-sectional area of pulmonary arterioles were significantly increased(P＜0.01).The positive expression rate of α-SMA protein in pulmonary artery was significantly increased(P＜0.01).The levels of plasma IL-1β,IL-6 and TNF-α were significantly increased(P＜0.01).The positive expression rate of PPAR-γ protein in lung tissue was significantly decreased(P＜0.01),and the positive expression rate of NF-κB protein was significantly increased(P＜0.01).The protein expressions of PPAR-γ and IκB-α in lung tissue were significantly down-regulated(P＜0.01).The protein expression ratio of p-NF-κB/NF-κB was significantly increased(P＜0.01).Compared with the model group,RVSP,mPAP,RVHI,RVIDd and RVAWT in each administration group were significantly decreased(P＜0.05,P＜0.01),while PAAT,PAAT/PET and TAPSE were significantly increased(P＜0.05,P＜0.01).The thickness of the vascular wall was significantly reduced,and the percentage of the wall thickness of the pulmonary arterioles to the diameter of the blood vessels and the percentage of the vascular wall area to the total cross-sectional area of the small arteries were significantly reduced(P＜0.05,P＜0.01).The positive expression rate of α-SMA protein in pulmonary artery was significantly decreased(P＜0.05,P＜0.01).The plasma levels of IL-1β,IL-6 and TNF-α were significantly decreased(P＜0.05,P＜0.01).The positive expression rate of PPAR-γ protein in lung tissue was significantly increased(P＜0.05,P＜0.01),and the positive expression rate of NF-κB protein was significantly decreased(P＜0.05,P＜0.01).The protein expression of PPAR-γ in lung tissue was significantly up-regulated(P＜0.05,P＜0.01),and the protein expression ratio of p-NF-κB/NF-κB was significantly decreased(P＜0.01).The protein expression of IκB-α in the lung tissue of rats in the high-dose group of Feixin Decoction was significantly up-regulated(P＜0.01).Conclusion Feixin Decoction can improve pulmonary artery pressure,right ventricular dysfunction and pulmonary vascular remodeling in PAH rats induced by monocrotaline.The mechanism may be related to the regulation of PPAR-γ/NF-κB signaling pathway to inhibit inflammatory response.

Pulmonary hypertension(PH)is a progressive pulmonary vascular disease that can lead to right heart failure and death.In recent years,the incidence of PH has been increasing year by year and there is a lack of effective treatment.TCM can play an important synergistic role in the treatment of PH.Pulmonary vascular remodeling is a core pathological feature of PH,which is closely related to the physiological structure and pathological changes of the collaterals.Based on the collateral disease theory,this article described the key pathogenesis of PH in TCM and Western medicine,including the lesions of the pulmonary and cardiovascular complexes and pulmonary vascular remodeling,analyzed the physiology of the"collateral-vessel"in PH,sorting out the pathological correlation,and explored TCM targeting pulmonary vascular remodeling in the identification and treatment of PH,so as to provide a new way of thinking for the clinical treatment of PH.

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.

Objective To monitor heart-related parameters of hypoxic pulmonary hypertension(PH)mouse models induced by hypoxia alone and hypoxia combined with vascular endothelial growth factor receptor inhibitor SU5416 using echocardiography,and to construct the prediction equation of right ventricular systolic pressure(RVSP).Twenty-four C57BL/6J male mice were randomly divided into simple hypoxia group(group A),hypoxia combined with SU5416 group(group B),control group(group C),each group 8 mice.Hypoxic PH models were constructed with hypoxia alone and hypoxia combined with SU5416 in group A and group B,respectively.Echocardiography was performed before and during modeling(2,3,4 weeks after interventions),and the relevant parameters were obtained.RVSP was measured using right heart catheterization after the last echocardiography.The changes of ultrasonic parameters were observed,the correlations of ultrasonic parameters 4 weeks after intervention with RVSP were observed,and linear equations for predicting RVSP were established.Results With time going,during modeling,pulmonary artery diameter(PAD),PAD/aorta diameter(AOD)and right ventricle anterior wall thickness(RVAWT)increased,while heart rate,pulmonary artery acceleration time(PAAT),PAAT/pulmonary artery ejection time(PAET)and tricuspid annular plane systolic excursion(TAPSE)decreased in group A and B(all P＜0.05).Three and 4 weeks after interventions,PAET,PAAT/PAET and TAPSE in group B decreased compared with those in group A(all P＜0.05).Four weeks after interventions,RVSP in group A and B were highly correlated with PAD/AOD,RVAWT,PAAT,PAAT/PAET and TAPSE(all P＜0.05).The linear regression equations of PAAT/PAET and TAPSE for predicting RVSP in simple hypoxic PH mice models included RVSP=-161.7 ×(PAAT/PAET)+63.85,as well as RVSP=-36.53 ×TAPSE+71.55,while of predicting RVSP in hypoxia combined with VEGFR-2 inhibitor PH mouse models were as follows:RVSP=-266.4 ×(PAAT/PAET)+91.59,RVSP=-69.14 × TAPSE+116.5.Conclusion Four weeks after inerventions,the phenotypes of hypoxic PH mouse models induced by hypoxia alone and hypoxia combined with SU5416 became obvious.Prediction equations of RVSP established based on PAAT/PAET and TAPSE obtained with echocardiography could provide references for relevant research.

Chronic obstructive pulmonary disease (COPD) is a common disease with high prevalence,high mortality and high disability.A large number of studies have confirmed that lung rehabilitation is an effective treatment for patients with COPD in addition to oxygen therapy and drug therapy.Respiratory muscle training is one of the basic methods of COPD lung rehabilitation,so do a good job in COPD patients with respiratory muscle training to improve the quality of life of patients,delay the disease process is of great significance.This paper focuses on the application progress of respiratory muscle training in COPD lung rehabilitation.

Objective To investigate the clinical efficacy of paraspinal muscle gap approach in the short segment internal fixation of thoracolumbar fractures.Methods Forty-five patients with thoraco-lumbar fractures but without spinal decompression underwent posterior short segment internal fixation,in-cluding 20 cases via paraspinal muscle gap approach(group A)and 25 cases via traditional posterior mid-line approach.Operative time,intraoperative blood loss,postoperative drainage,visual analogue scale (VAS)score,correction rate of Cobb angle and correction rate of vertebral collapse were recorded and the surgical results were compared between the groups.Results All patients got bony union without internal fixation loosening,broken nails or broken rods.There were significant differences in operative time [(88 ± 17)min vs(105 ±14)min],blood loss [(121 ±24)ml vs(230 ±31 )ml]and postoperative drainage [(66 ±28)ml vs(250 ±45)ml]between group A and B respectively(P<0.05).The differences in cor-rection rate of the Cobb angle [(82.3 ±1.58)% vs(83.5 ±3.71)%],correction rate of the vertebral collapse [(88.22 ±3.18)%vs(87.19 ±2.16)%]and postoperative VAS score were not significant be-tween group A and B respectively(P>0.05 ).Conclusion Internal fixation via paraspinal muscle gap approach has the advantages of less trauma,simple approach,short operative time,less blood loss,quick recovery after surgery.It is consistent with the modern concept of minimal invasion and worthy of sprea-ding.

As one of the main members of the Rho GDI dissociation inhibitory factors,D4-GDI inhibits the dissociation of Rho protein and GDP,which is also involved in a wide range of celluar functions,such as cell contraction,adhesion,migration,proliferation and apoptosis.Recently,accumulating evidence has been suggested that D4-GDI is involved in the pathogenesis of several pulmonary diseases,such as lung cancer.Intervention of D4-GDI expression may improve the pathological changes and prognosis of these diseases.