Secondary organizing pneumonia after infection is a pathological condition characterized by connective tissue filling and obstructing the alveoli and bronchioles, in which following an infection in the lung, the inflammatory response is not controlled in a timely and effective manner. The pathogenesis and treatment of this condition can be interpreted through the Sanjiao membranous tube theory and the concept of stagnation within the pulmonary micro-membrane. Sanjiao is conceptualized as a four-way membranous tube that internally connects with the zangfu organs and externally with the skin and muscles, enabling the circulation of energy and fluids throughout the body. It also maintains communication with the zangfu micro-membranes. Within the lungs, the pulmonary micro-membrane is distributed and connected to the upper jiao membranous tube, facilitating the movement of qi and fluids and supporting nutrient distribution. External pathogens may invade the Sanjiao membranous system through the external membranous tube, travel internally along this system, and transform into latent pathogens that settle within the pulmonary micro-membrane. These latent pathogens can subsequently transform into heat or dampness, leading to the depletion of lung qi and impairing the lung′s ability to regulate and transport body fluids. Consequently, fluids may seep into the pulmonary micro-membrane, where they are transformed into dampness, turbidity, and phlegm. The accumulation of damp-turbidity and phlegm obstructs the flow of qi and blood, resulting in blood stasis in the pulmonary collaterals. This stagnation occurring within both the pulmonary micro-membrane and its associated collaterals underlies the development of secondary organizing pneumonia after infection. In severe cases, this condition may progress to pulmonary interstitial fibrosis. The therapeutic approach emphasizes expelling latent pathogens, regulating and dredging the pulmonary micro-membrane, tonifying the healthy qi, and supporting health. Regulating and dredging the pulmonary micro-membrane is a crucial step, with a focus on promoting the flow of lung qi, resolving dampness and phlegm, and activating blood circulation to remove stasis.

OBJECTIVE: To observe the effects of acupoint catgut embedding (ACE) on gut microbiota and fecal short-chain fatty acids (SCFAs) levels in patients with Parkinson's disease (PD) with constipation. METHODS: A total of 80 PD patients with constipation were randomly divided into an observation group and a control group, 40 cases in each group. Additionally, 40 healthy individuals were recruited as a healthy control group. The control group received conventional Western medical treatment for PD combined with polyethylene glycol (PEG), once daily for eight weeks. The observation group received additional ACE treatment at bilateral Tianshu (ST25), Zusanli (ST36), and Shangjuxu (ST37), once every two weeks for eight weeks. The healthy control group received no intervention. The spontaneous bowel movements (SBMs) per week and patient assessment of constipation quality of life (PAC-QOL) scores were assessed at baseline and after treatment in the two groups. Fecal samples were collected at the end of treatment for the observation and the control groups and at baseline for the healthy control group. Gut microbiota composition and diversity were analyzed using 16S rRNA method, and SCFA levels were measured using high-performance liquid chromatography (HPLC). RESULTS: Compared before treatment, the observation group showed a significant increase in SBMs (P<0.01), and PAC-QOL scores including physical discomfort, psychosocial discomfort, worry and concern, and total score were significantly reduced (P<0.01) after treatment; the control group also showed a reduction in PAC-QOL total score after treatment (P<0.01). After treatment, the observation group had significantly more SBMs (P<0.01), and lower PAC-QOL physical discomfort, psychosocial discomfort, worry and concern scores, and total score (P<0.01), and higher PAC-QOL satisfaction score (P<0.01) than the control group. Compared with the healthy control group, the control group showed decreased Chao1 and Ace indices (P<0.01). Compared with the healthy control group, the relative abundance of Prevotella and Roseburia was increased (P<0.05), while that of Enterobacter and Ruminococcus torques (six species in total) was decreased (P<0.05) in the control group. Compared with the control group, the observation group had increased relative abundance of Dialister, Parabacteroides, and Ruminococcus torques (P<0.05), and decreased relative abundance of Prevotella and Eubacterium ruminantium (P<0.05). Compared with the healthy control group, the control group had increased fecal SCFA levels (P<0.05); compared with the control group, the observation group had reduced fecal SCFA levels (P<0.05). Compared with the healthy control group, acetic acid, propionic acid, and butyric acid levels were elevated in the control group (P<0.05); compared with the control group, acetic acid, propionic acid, and butyric acid levels were decreased in the observation group (P<0.05). CONCLUSION ACE could increase spontaneous bowel movements and improve the quality of life in PD patients with constipation, which may be related to the regulation of gut microbiota composition and SCFA levels.
Humans ; Constipation/metabolism* ; Male ; Gastrointestinal Microbiome ; Acupuncture Points ; Female ; Middle Aged ; Parkinson Disease/complications* ; Aged ; Fatty Acids, Volatile/metabolism* ; Catgut ; Feces/microbiology* ; Acupuncture Therapy ; Quality of Life ; Adult

Objective:To construct a sizeable naive human Fab phage display antibody library to screen high-affinity specific antibodies in vitro. Methods:Total RNA was extracted from peripheral blood mononuclear cells (PBMCs) of 126 healthy individuals, subsequently reverse-transcribed into cDNA, and used as a template. PCR amplification was performed to obtain the V H from IgG, IgM and light chain κ, λ, separately, with the initial PCR products serving as templates for a second round of PCR. Overlap extension PCR was employed to generate fragments of the κ and λ light chains. These fragments were ligated with the phage vector pNC3, which harbors the variable region 1 of the heavy chain, to construct a recombinant phage plasmid. This plasmid was then electroporated into competent Escherichia Coli TG1 cells to establish a naive human Fab phage display antibody library. One hundred clones were randomly selected for identification and sequencing, and antibody gene polymorphisms were analyzed using the IMGT database and MAFFT software. Recombinant α-hemolysin from Staphylococcus aureus was utilized to screen Fab antibody fragments through biopanning of the antibody library, followed by random selection of phage ELISA-identified clones. The positive clones (antigen A450∶blank control A450≥2.1) were sequenced. Results:Two large naive Fab phage display antibody libraries were successfully constructed, in which the capacity of κ and λ chain antibody libraries were 1.25×10 11 and 1.54×10 11, respectively. The titers for two antibody libraries were 6.04×10 13 CFU/ml and 3.50×10 13 CFU/ml. The positive transformation insertion rates for κ and λ chain antibody libraries were 96% (96/100) and 100% (100/100), respectively. Sequence analysis revealed that all antibody sequences were unique. The amino acid sequences in the skeletal region were relatively conserved. In contrast, significant variations in the length of the complementarity determining region (CDR) were found, and the diversity of amino acid sequence of the complementary determining region was high, especially the CDR3. Analysis using the IMGT database indicated that the sequences exhibited a broad distribution across variable-diversity-joining gene families. After six rounds of panning, specific phage antibodies enrichment targeting α-hemolysin were achieved. A total of 142 monoclonal antibodies were sequenced, yielding 8 distinct Fab antibody sequences. Conclusion:This study successfully constructed two naive human Fab phage display antibody libraries with large capacity and good diversity, which can be used for screening human antibodies for serum epidemiology.

Objective:To analyze the clinical characteristics and treatment outcomes of children with anti-3-hydroxy-3-methylglutaryl-coenzyme A reductase (anti-HMGCR) antibody-mediated necrotizing myopathy, and to explore early identification and management strategies to provide reference for clinical diagnosis and treatment.Methods:A retrospective analysis was conducted on the clinical data and treatment outcomes of 10 pediatric patients with anti-HMGCR antibody-mediated necrotizing myopathy admitted to the Department of Rheumatology, Children′s Hospital of Fudan University from December 2020 to December 2024. Statistical description was performed using SPSS 22.0.Results:Among the 10 patients, the male-to-female ratio was 1:4, the age of onset was (7.2±4.0) years, and the disease duration at diagnosis was (22.2±19.6) months. None had a history of statin exposure. Six patients presented with muscle weakness, and4 were diagnosed due to asymptomatic elevation of creatine kinase (CK); 4 had dermatomyositis-like rashes. All patients showed significantly elevated CK levels [median 3 291(1 969, 8 776)U/L] and underwent muscle biopsy. Histopathological findings revealed myofiber degeneration, necrosis, and regeneration in all cases, with inflammatory infiltration in 9 cases, MHC-Ⅰ positivity in all, and C5b-9 positivity in 9 cases. The median follow-up duration was (15.7±6.3) months. At the last follow-up, muscle strength was normal or nearly normal, and the CK median value had decreased to 977.5 (211.0, 3 536.0) U/L.Conclusion:For patients with suspected idiopathic inflammatory myopathy and significantly elevated CK, muscle-specific antibody testing-including anti-HMGCR-and muscle biopsy should be performed promptly regardless of the presence of skin rash, to ensure accurate diagnosis and guide treatment, thereby avoiding misdiagnosis or missed diagnosis.

This study investigates the feasibility of the VP8*protein as a subunit vaccine target for porcine rotavirus A(PoRVA),a major causative agent of diarrhea in piglets.The VP8* genes of PoRVA P[13]and P[23]genotype strains were amplified by RT-PCR.These genes were then liga-ted into the pET-28a(+)vector,yielding recombinant plasmids pET-28a-XJWF1-VP8*-P[23]and pET-28a-ShXYW13-VP8*-P[13].These plasmids were subsequently transformed into BL21(DE3)competent cells.The VP8*protein,induced by IPTG,was purified using affinity chroma-tography,and its expression and purification were verified by SDS-PAGE and Western blot.The purified VP8* protein was used to immunize mice,and serum samples were collected after three immunizations.Cross-neutralization assays were conducted to evaluate the ability of the VP8*protein immune serum to neutralize different genotype strains.The results demonstrated the ex-pression of soluble VP8*protein,with SDS-PAGE and Western blot analyses showing that the purified VP8*protein existed in both monomeric(27 kDa)and homodimeric(54 kDa)forms.ELISA results indicated that high levels of antibodies were produced in mice immunized with VP 8*-P[13]and VP8*-P[23]after three immunizations.Serum cross-neutralization assays revealed that the neutralizing titers of PoRVA VP8*-P[13]and VP8*-P[23]immune sera against homol-ogous genotype strains ranged from 1∶4 800 to 1∶19 200,significantly higher than those against heterologous genotype strains(1∶1 200).This suggests that the VP8*protein of different geno-type strains exhibits both antigenic conservation and distinct variability.The data obtained in this study provide a solid foundation for further exploration of the antigenic structure of the PoRVA VP8* protein and the development of novel subunit vaccines.

Objective·To investigate the genetic etiology of a rare and complex case clinically suspected to be methylmalonic acidemia(MMA),but with negative whole exome sequencing(WES)results,using a multi-omics sequencing approach.Methods·DNA and RNA samples were extracted from the peripheral blood of the proband and both parents.Targeted MMA-related gene Panel sequencing and WES were first performed.Subsequently,RNA sequencing(RNA-seq)and whole genome sequencing(WGS)were conducted to comprehensively analyze the child's genetic variants,their origins and potential inheritance patterns.Results·No pathogenic variants associated with the patient's phenotype were identified through the MMA Panel or standard WES analysis.Extended analysis of WES suggested the possibility of uniparental disomy(UPD)of chromosome 4.WGS revealed a homozygous splice-site variant(c.-66+2T>C)in the non-coding region of the metabolism of cobalamin associated A(MMAA)gene.The variant was located in the 5'untranslated region(5'UTR),specifically at the second base downstream of the splice donor site of exon 1(reference sequence:NM_172250).In genomic coordinates(hg19),the variant was located at base 146540561 on chromosome 4(chr4:146540561).Sanger sequencing confirmed that the mother was heterozygous for this variant,while the father did not carry it.RNA-seq showed no detectable expression of the MMAA gene on chromosome 4 in the patient.This was further confirmed by reverse transcription real time quantitative PCR,indicating nearly absent mRNA expression,suggesting that the non-coding splice-site variant affected transcriptional expression.Conclusion·A homozygous splice-site variant(c.-66+2T>C)in the non-coding region of the MMAA gene—outside the coverage of WES—is likely the pathogenic cause in this case,presumably resulting from maternal UPD of chromosome 4.

Systemic sclerosis is an autoimmune rheumatic disease that often leads to multisystem diseases,frequently resulting in pulmonary interstitial fibrosis.According to the theory of sanjiao(triple energizers)membranous channels,sanjiao connects the five zang-organs and six fu-viscera internally and the skin,muscles,and bones externally.It serves as a four-way membranous channel that connects internal organs and external structures,linking with the micromembranes of organs and blood vessels.The pathogenesis of pulmonary interstitial fibrosis secondary to systemic sclerosis involves external cold obstructing the skin and interstitial layers,impairing the defense qi and defense yang,which originate in the essence of the kidney.This leads to weak defensive qi and kidney deficiency,causing stagnation in sanjiao's energy flow and disruption of water and gasification and loss of fluid,resulting in accumulation of dampness,phlegm,and blood stasis.These obstructive factors spread along sanjiao's membranous channels,leading to multiorgan micromembrane involvement and systemic damage.The lungs,which are in direct contact with the external environment,are particularly susceptible to invasion by external pathogens.When combined with stagnation of dampness,phlegm,and blood in the lungs,this leads to secondary pulmonary fibrosis,resulting in lung dysfunction.Continuous stagnation of sanjiao exacerbates the overall condition of the patient,leading to a mixed cold-heat imbalance.Treatment focuses on"unblocking,transforming,and regulating"to restore sanjiao function,promote qi and fluid circulation,invigorate blood,and adjust the cold-heat imbalance,ultimately restoring the overall condition of the patient.

Objective To observe the efficacy and safety of Yifei Sanjie Decoction in the treatment of multiple pulmonary nodules.Methods A prospective randomized controlled clinical study was conducted to select 189 patients with multiple pulmonary nodules who saught medical attention at the Pulmonary Nodule Diagnosis and Treatment Center of Dongfang Hospital,Beijing University of Chinese Medicine and the Department of Thoracic Surgery of Beijing Shijitan Hospital Affiliated to Capital Medical University from January 2023 to March 2025.According to the random number table method,126 cases were randomly divided into the trial group and 63 cases in the blank control group at a ratio of 2∶1.The trial group was treated with modified Yifei Sanjie Decoction,and the blank control group was only followed up without intervention.The course of treatment was 3 months.The patients in the two groups were reexamined with lung CT after 3 months.The efficacy was evaluated by the area reduction rate of the major pulmonary nodules and the cumulative multiple pulmonary nodules in patients with multiple pulmonary nodules combined with the average diameter and malignant signs.According to the property of the major pulmonary nodules,we divided the patients into ground glass,solid,and mixed ground glass subgroups to evaluate the efficacy of different types of pulmonary nodules.We evaluated the change of malignant risk of pulmonary nodules according to the change of Mayo score.We evaluated the safety of the treatment medicine by blood routine,urine routine,and liver and kidney function.Results A total of 175 patients completed the study,117 in the trial group and 58 in the blank control group.The total effective rates of the major pulmonary nodules and the cumulative multiple pulmonary nodules in the trial group were 41.03%and 42.74%,respectively,the total effective rate of nodule number change was 29.91%which were significantly higher than those in the blank control group(P<0.05).In the trial group,the total effective rates of the major pulmonary nodules in the ground glass(72 cases),solid(28 cases),and mixed ground glass(17 cases)subgroups were 40.28%,32.14%,and 58.82%,respectively,the total effective rates of the cumulative multiple pulmonary nodules were 38.89%,42.86%,and 58.82%,respectively,which were significantly higher than those in corresponding subgroups of ground glass(37 cases),solid(14 cases),and mixed ground glass(7 cases)of the blank control(P<0.05).After treatment,the average diameter,area,and Mayo score of the major pulmonary nodules and the cumulative multiple pulmonary nodules in the trial group were significantly lower than those in the blank control group and before treatment(P<0.05),the average diameter,area,and Mayo score of the major pulmonary nodules and the cumulative multiple pulmonary nodules in the ground glass,solid,and mixed ground glass subgroups were lower than those in corresponding subgroups of the blank control group,and significantly lower than those before treatment(P<0.05).The number of nodules and malignant signs in the trial group were lower than before,while those in the blank control group were higher than before.There were no serious adverse events in the two groups during the study.Conclusion Yifei Sanjie Decoction can effectively treat the major pulmonary nodules and the cumulative multiple pulmonary nodules in patients with multiple pulmonary nodules,reduce the average diameter and area of nodules,reduce the Mayo score,and reduce the malignant signs and number of nodules.In the ground glass,solid,and mixed ground glass groups,the curative effect is well,and the safety is high,it can be used for the clinical treatment of multiple pulmonary nodules.

Objective To investigate the relationship between triglyceride glucose body mass index(TyG-BMI),high density lipoprotein cholesterol to Apolipoprotein A ratio(HAR),glycemic risk index(GRI)and diabetic retinopathy(DR).Methods A total of 159 patients with type 2 diabetes mellitus(T2DM)complicated with DR(the DR group)were divided into the non-proliferative retinopathy group(NPDR group,66 cases)and the proliferative retinopathy group(PDR group,93 cases)according to DR international clinical grading criteria,and 159 T2DM patients without DR were selected as the control group.Clinical information and baseline laboratory test results were recorded,and TyG-BMI,HAR and GRI were calculated.Multivariate Logistic regression analysis was conducted to analyze the related factors of the incidence of DR,and receiver operating characteristic(ROC)curve was used to analyze the diagnostic value of TyG-BMI,HAR and GRI for DR.Results The duration of T2DM,the proportion of hypertension,diabetic nephropathy and diabetic foot,levels of HbA1c,homeostasis model insulin resistance index(HOMA-IR),total cholesterol(TC),C-reactive protein(CRP),TyG-BMI,HAR and GRI were higher in the DR group than those in the control group(P<0.05).TyG-BMI,HAR and GRI were higher in the PDR group than those in the NPDR group(P<0.05).Multivariate Logistic regression analysis showed that the longer course of T2DM disease[OR(95%CI):2.781(1.398-5.534)],high TyG-BMI[2.036(1.169-3.546)],high HAR[1.890(1.090-3.280)]and high GRI[1.836(1.065-3.167)]were risk factors for DR(P<0.05).ROC curve analysis results showed that,the area under the curve(AUC)of combined TyG-BMI,HAR and GRI diagnosis of DR was higher than that of single diagnosis[0.940(0.908-0.964),0.864(0.821-0.900),0.796(0.747-0.839)and 0.836(0.790-0.875),all P<0.05].Conclusion The increased TyG-BMI,HAR and GRI in T2DM patients is associated with the onset and severity of DR,and which can be used to assess the risk of DR.