ObjectiveTo investigate the intervention mechanism of the traditional Chinese medicine Number 2 Feibi recipe （N2FBR） in idiopathic pulmonary fibrosis （IPF）， focusing on its effects on endoplasmic reticulum （ER） stress， apoptosis， stemness maintenance， and regenerative capacity of alveolar type Ⅱ epithelial cells （AT2 cells）， and to validate the modern translational pathway of the theory of "deficiency of Zong Qi leading to pulmonary atelectasis and atrophy". MethodsA mouse model of pulmonary fibrosis was induced by bleomycin （BLM）. Mice were randomly divided into blank control， model， low-， and high-dose N2FBR intervention groups （9.1， 18.2 g·kg^-1）， and prednisolone intervention group （6.5 mg·kg^-1）. Pulmonary histopathological changes and collagen deposition were evaluated using hematoxylin-eosin （HE） and Masson's trichrome staining. Hydroxyproline （HYP） content was measured by the alkaline hydrolysis method. Lung coefficient and pulmonary function parameters were evaluated. The mRNA expression levels of fibrosis-related factors， including collagen type Ⅰ alpha 1 chain （ColIa1）， alpha-smooth muscle actin （α-SMA）， and tissue inhibitor of metalloproteinase 1 （Timp1）， were detected by real-time polymerase chain reaction （Real-time PCR）. Cell apoptosis was assessed using the terminal deoxynucleotidyl transferase dUTP nick-end labeling （TUNEL） assay. Apoptosis of AT2 cells was further evaluated by double immunofluorescence staining for surfactant protein C （SPC） and cysteine-aspartic protease-3 （Caspase-3）. Endoplasmic reticulum （ER） stress in AT2 cells was examined by double staining for SPC and protein kinase R-like endoplasmic reticulum kinase （PERK）. Ultrastructural changes of ER and lamellar bodies in AT2 cells were observed by transmission electron microscopy （TEM）. The expression levels of key proteins involved in ER stress and apoptosis pathways， including PERK， activating transcription factor 4 （ATF4）， and Caspase-3， were detected by Western blot. Double immunofluorescence staining of SPC and Ki-67 antigen （Ki-67） was performed to evaluate the proliferative capacity of AT2 cells. Lineage tracing technology （labeling AT2 cells with GFP） combined with Krt8 labeling was used to evaluate intermediate differentiation states， and morphological transformation of AT2 cells into alveolar type Ⅰ epithelial cells （AT1） was observed. ResultsBLM-induced mice exhibited significant structural disruption of lung tissue， increased collagen deposition， elevated lung coefficient， decreased pulmonary function， and upregulation of fibrosis-related factors （P<0.01）. High-dose N2FBR treatment significantly ameliorated lung tissue damage and dysfunction， significantly reduced HYP content （P<0.01）， and significantly downregulated ColIa1， α-SMA， and Timp1 expression （P<0.01）. Apoptosis analysis showed increased TUNEL-positive and Caspase-3-positive AT2 cells in the model group， which was significantly reduced by high-dose N2FBR treatment. TEM revealed swollen ER structures in AT2 cells of the model group， which tended to return to normal following treatment. PERK protein staining analysis showed evident ER stress in AT2 cells of the model group， which were markedly alleviated in the treatment group. The expression levels of ER stress-related proteins PERK and ATF4， as well as the apoptosis-related protein Caspase-3， were elevated in the model group and significantly reduced after treatment. TEM also revealed disrupted lamellar body structures in the model group， which tended to recover in the treatment group. Regarding the proliferative capacity of AT2 cells， the proportion of Ki-67⁺SPC⁺ AT2 cells significantly increased in the treatment group （P<0.01）. Lineage tracing showed that the proportion of keratin 8-positive green fluorescent protein-positive （Krt8⁺GFP⁺） cells increased in the model group， indicating differentiation arrest. This proportion was significantly reduced in the treatment group， and the morphology of GFP⁺ cells exhibited a flattened， extended shape， suggesting restored differentiation toward AT1 cells. ConclusionN2FBR alleviates ER stress in AT2 cells， reduces AT2 cell apoptosis， restores lamellar body structure and function， enhances proliferation activity， and alleviates differentiation arrest to promote differentiation into AT1 cells， thereby repairing the alveolar epithelium and effectively blocking the progression of pulmonary fibrosis. Its traditional Chinese medicine mechanism of "replenishing Zong Qi， harmonizing Qi and blood， and unblocking pulmonary meridians" closely aligns with the modern regulatory pathway of AT2 stem cells， providing a novel theoretical basis and experimental evidence for the intervention of IPF with traditional Chinese medicine.

ObjectiveTo investigate the intervention mechanism of the traditional Chinese medicine Number 2 Feibi recipe （N2FBR） in idiopathic pulmonary fibrosis （IPF）， focusing on its effects on endoplasmic reticulum （ER） stress， apoptosis， stemness maintenance， and regenerative capacity of alveolar type Ⅱ epithelial cells （AT2 cells）， and to validate the modern translational pathway of the theory of "deficiency of Zong Qi leading to pulmonary atelectasis and atrophy". MethodsA mouse model of pulmonary fibrosis was induced by bleomycin （BLM）. Mice were randomly divided into blank control， model， low-， and high-dose N2FBR intervention groups （9.1， 18.2 g·kg^-1）， and prednisolone intervention group （6.5 mg·kg^-1）. Pulmonary histopathological changes and collagen deposition were evaluated using hematoxylin-eosin （HE） and Masson's trichrome staining. Hydroxyproline （HYP） content was measured by the alkaline hydrolysis method. Lung coefficient and pulmonary function parameters were evaluated. The mRNA expression levels of fibrosis-related factors， including collagen type Ⅰ alpha 1 chain （ColIa1）， alpha-smooth muscle actin （α-SMA）， and tissue inhibitor of metalloproteinase 1 （Timp1）， were detected by real-time polymerase chain reaction （Real-time PCR）. Cell apoptosis was assessed using the terminal deoxynucleotidyl transferase dUTP nick-end labeling （TUNEL） assay. Apoptosis of AT2 cells was further evaluated by double immunofluorescence staining for surfactant protein C （SPC） and cysteine-aspartic protease-3 （Caspase-3）. Endoplasmic reticulum （ER） stress in AT2 cells was examined by double staining for SPC and protein kinase R-like endoplasmic reticulum kinase （PERK）. Ultrastructural changes of ER and lamellar bodies in AT2 cells were observed by transmission electron microscopy （TEM）. The expression levels of key proteins involved in ER stress and apoptosis pathways， including PERK， activating transcription factor 4 （ATF4）， and Caspase-3， were detected by Western blot. Double immunofluorescence staining of SPC and Ki-67 antigen （Ki-67） was performed to evaluate the proliferative capacity of AT2 cells. Lineage tracing technology （labeling AT2 cells with GFP） combined with Krt8 labeling was used to evaluate intermediate differentiation states， and morphological transformation of AT2 cells into alveolar type Ⅰ epithelial cells （AT1） was observed. ResultsBLM-induced mice exhibited significant structural disruption of lung tissue， increased collagen deposition， elevated lung coefficient， decreased pulmonary function， and upregulation of fibrosis-related factors （P<0.01）. High-dose N2FBR treatment significantly ameliorated lung tissue damage and dysfunction， significantly reduced HYP content （P<0.01）， and significantly downregulated ColIa1， α-SMA， and Timp1 expression （P<0.01）. Apoptosis analysis showed increased TUNEL-positive and Caspase-3-positive AT2 cells in the model group， which was significantly reduced by high-dose N2FBR treatment. TEM revealed swollen ER structures in AT2 cells of the model group， which tended to return to normal following treatment. PERK protein staining analysis showed evident ER stress in AT2 cells of the model group， which were markedly alleviated in the treatment group. The expression levels of ER stress-related proteins PERK and ATF4， as well as the apoptosis-related protein Caspase-3， were elevated in the model group and significantly reduced after treatment. TEM also revealed disrupted lamellar body structures in the model group， which tended to recover in the treatment group. Regarding the proliferative capacity of AT2 cells， the proportion of Ki-67⁺SPC⁺ AT2 cells significantly increased in the treatment group （P<0.01）. Lineage tracing showed that the proportion of keratin 8-positive green fluorescent protein-positive （Krt8⁺GFP⁺） cells increased in the model group， indicating differentiation arrest. This proportion was significantly reduced in the treatment group， and the morphology of GFP⁺ cells exhibited a flattened， extended shape， suggesting restored differentiation toward AT1 cells. ConclusionN2FBR alleviates ER stress in AT2 cells， reduces AT2 cell apoptosis， restores lamellar body structure and function， enhances proliferation activity， and alleviates differentiation arrest to promote differentiation into AT1 cells， thereby repairing the alveolar epithelium and effectively blocking the progression of pulmonary fibrosis. Its traditional Chinese medicine mechanism of "replenishing Zong Qi， harmonizing Qi and blood， and unblocking pulmonary meridians" closely aligns with the modern regulatory pathway of AT2 stem cells， providing a novel theoretical basis and experimental evidence for the intervention of IPF with traditional Chinese medicine.

BackgroundPrenatal depression has an important impact on maternal health and pregnancy outcomes. Previous studies have shown that maternal prenatal depression is associated with social support， and social support is related to fear of childbirth. However， there is limited research on the relationship among maternal prenatal depression， social support and fear of childbirth， and no studies have specifically explored the influence of social support and fear of childbirth on prenatal depression in primiparous women. ObjectiveTo investigate the current status of prenatal depression among primiparous women， and to analyze the correlation between social support and fear of childbirth， and to further explore the influence of social support and fear of childbirth on prenatal depression in this population， so as to provide references for improving their mental health. MethodsA total of 380 primiparous women admitted to the inpatient department of Chengdu Wenjiang District People's Hospital from December 2022 to September 2023 were enrolled as study subjects. A self-made questionnaire， Edinburgh Postnatal Depression Scale （EPDS）， Social Support Rating Scale （SSRS） and Childbirth Attitudes Questionnaire （CAQ） were used to conduct the survey. Pearson correlation analysis was employed to examine the relationships between scale scores. Multiple linear regression analysis was conducted to identify influencing factors of prenatal depression. ResultsA total of 380 questionnaires were distributed， with 372 （97.89%） valid responses collected. Among the participants， 222 cases （59.68%） were identified with prenatal depression. Pearson correlation analysis revealed that EPDS score was negatively correlated with SSRS score （r=-0.283， P<0.01） and positively correlated with CAQ score （r=0.341， P<0.01）. Multiple linear regression analysis indicated that social support （β=-0.166， P<0.01） and fear of childbirth （β=0.269， P<0.01） were influencing factors of prenatal depression in primiparous women. ConclusionThe prevalence of prenatal depression among primiparous women is concerning， with depression levels showing significant associations with both social support and fear of childbirth.

Objective To identify potential serum metabolic biomarkers in colorectal cancer(CRC)patients using untargeted metabolomics and to evaluate their diagnostic and staging value.Methods Serum samples from 100 healthy controls and 100 CRC patients were analyzed by ultra-performance liquid chromatography-mass spectrometry(UPLC-MS).After data normalization,differential metabolites were screened using multivariate statistical analyses(PCA,OPLS-DA)and subjected to pathway enrichment analysis.Diagnostic performance was assessed via univari-ate and multivariate regression,while Mfuzz clustering was applied to analyze stage-related metabolites(Ⅰ-Ⅳ).Results A total of 432 metabolites were identified with 59 showing significant alterations.Starch and sucrose me-tabolism and glycerophospholipid metabolism pathways were significantly enriched.A three-metabolite panel(4,8-dimethylnonanoyl carnitine,9,13-dihydroxy-4-megastigmen-3-one 9-glucoside and C17 sphingosine-1-phosphate)achieved a diagnostic AUC of 0.907,while L-Carnitine and L-Norleucine showed an AUC of 0.776 in staging anal-ysis.Conclusions Specific serum metabolite panel exhibit high diagnostic accuracy,and dysregulated metabolic pathways are associated with CRC progression,suggesting their potential value as biomarkers.

Objective To observe the clinical efficacy of dynamic joint mobilization combined with core stability training in the treatment of nonspecific low back pain.Methods 60 patients with nonspecific low back pain were randomly assigned into either the treatment group or control group.Each group had 30 patients.The treatment group received a therapeutic regimen combining dynamic joint mobilization and core stability training,while the control group only received core stability training.Both groups were evaluated for therapeutic effectiveness using the Visual Analogue Scale(VAS)for pain,the Oswestry Disability Index(ODI),and the range of motion(ROM)of the lumbar spine before treatment,and at the 1st,3rd,and 6th weeks after treatment.At the conclusion of the treatment,a thorough assessment of the overall therapeutic efficacy was performed.Results At the 1st,3rd,and 6th weeks post-treatment,both groups showed statistically significant differences in VAS scores,ODI scores,and ROM scores over time(P＜0.05).The improvements in these indices were significantly greater in the treatment group compared to the control group(P＜0.05).The treatment group had considerably higher therapeutic effective-ness compared to the control group(P＜0.05).Conclusions Dynamic joint mobilization combined with core stabil-ity training is effective in treating nonspecific low back pain.It can help with pain relief,lumbar and back function restoration,and lumbar and back mobility improvement.This approach is worthy of clinical application and promotion.

Objective:To explore the clinical phenotype and genetic etiology of a child with Developmental epileptic encephalopathy type 104 (DEE 104).Methods:A child who had presented at the Children′s Medical Center of the Affiliated Hospital of Guangdong Medical University in February 2021 for recurrent seizures over 1 month was selected as the study subject. Clinical data of the child was collected. Peripheral blood samples of the child and his parents were collected and subjected to whole exome sequencing (WES). Candidate variant was verified by Sanger sequencing.Results:The child, a five-month-old male, had presented with frequent focal seizures with severe developmental retardation from infancy. Physical examination showed emaciation, microcephaly, oblique palpebral fissures, Stahl′s ears, and hypotonia in the limbs. Electroencephalogram revealed multi-focal sharp waves, slow waves and slow spinal waves. Cranial magnetic resonance imaging revealed enlargement of bilateral lateral ventricles and the third ventricle, along with widening of brain sulci, fissure and cisterna. WES revealed that he had harbored a heterozygous c. 2401C>T (p.His801Tyr) missense variant of the ATP6V0A1 gene. Sanger sequencing showed that both of his parents were of the wild type. Based on the guidelines from the American College of Medical Genetics and Genomics (ACMG), the variant was predicted to be likely pathogenic (PS2+ PM2_Supporting+ PP3). The proband was diagnosed with DEE 104. Early treatment with sodium valproate has failed, but the child had become seizure free after the addition of levetiracetam and topiramate. He still had abnormal EEG discharges and severe psychomotor retardation. Combining our case and a review of literature, DEE104 is mainly caused by de novo heterozygous variants of the ATP6V0A1 gene with an autosomal dominant inheritance. The patients may show refractory epilepsy and severe global developmental delay from infancy. Conclusion:The c. 2401C>T (p.His801Tyr) variant probably underlay the DEE104 in this child.

Objective:To explore the clinical features and genetic basis for a child with Intellectual developmental disorder (IDD) and epilepsy.Methods:A child who was admitted to the Children′s Medical Center of the Affiliated Hospital of Guangdong Medical University in February 2021 was selected as the study subject. Clinical data of the child was collected. Peripheral blood samples of the child and her parents were collected and subjected to whole exome sequencing (WES). Candidate variant was verified by Sanger sequencing.Results:The patient, a 3-month-and-27-day female infant, had developed the symptoms in the neonatal period, which included severe developmental delay, respiratory difficulties and pauses, increased muscle tone of four limbs, feeding difficulty, and seizures. Cerebral MRI revealed bilateral cerebellar hypoplasia, and video EEG showed slightly increased sharp waves emanating predominantly from the right parietal, occipital, and posterior temporal regions. WES revealed that she has harbored a missense c. 3196G>A (p.Glu1066Lys) variant of the CLTC gene, which was confirmed to be de novo by Sanger sequencing. Based on the guideline from the American College of Medical Genetics and Genomics (ACMG), the variant was classified as likely pathogenic (PS2+ PM2_Supporting+ PP3). Conclusion:The c. 3196G>A (p.Glu1066Lys) missense variant of the CLTC gene probably underlay the pathogenesis in this child. Above finding has facilitated her diagnosis and treatment.

Objective:To explore the influencing factors of early postoperative complications after radical resection of congenital choledochal cyst (CCC) in a single center and provide some clinical basis and guidance for reducing postoperative complications.Methods:Case control study.Clinical data of 124 children (29 boys and 95 girls) with CCC diagnosed and radically treated at the Affiliated Hospital of Zunyi Medical University from September 2010 to October 2019 were analyzed.According to postoperative complications (bile leakage, gastrointestinal anastomotic fistula, bleeding, incision dehiscence, cholangitis, abdominal infection, pancreatitis, and lymphatic fistula), these children were divided into the complication group (group A) and non-complication group (group B). Age, laboratory indicators[preoperative white blood cell (WBC) count, hemoglobin, glutamic pyruvic transaminase, prealbumin, and postoperative albumin], and clinical factors, such as operation method, operation time, intraoperative blood loss, cyst type, cyst diameter, hepatic duct diameter, abdominal operation history, biliary sludge and calculus, hepatic duct anatomic variation, and pancreaticobiliary maljunction were statistically analyzed between the two groups.The t-test was performed for normal distribution of the measurement data, and the non-parametric rank sum test for non-normal distribution.Multivariate analysis was made using Logistic regression. Results:Among the 124 children, 25(20.16%) had complications, and 99(79.84%) had no complications.Bile leakage occurred in 14 children (11.29%), of whom 7 received operation again and 7 received conservative treatment.Gastrointestinal anastomotic fistula occurred in 2 children (1.61%), of whom 1 was re-operated and 1 was cured conservatively.One child (0.81%) was complicated with bleeding and cured by re-operation.Two children (1.61%) were complicated with incision dehiscence, of whom 1 was cured by re-operation and 1 was cured by conservative treatment.Cholangitis in 2 children (1.61%), abdominal infection in 2 children (1.61%), pancreatitis in 1 child (0.81%), and lymphatic fistula in 1 child (0.81%) were all conservatively cured.No significant difference was found in non-normal distribution indicators-age and WBC count-between the two groups (all P>0.05). Blood loss volume and cyst diameter were significantly different between the two groups (all P<0.05). Postoperative albumin[(27.84±4.62) g/L vs.(32.45±3.72) g/L] meeting the normal distribution showed a statistically significant difference between the two groups ( t=5.254, P<0.05). Logistic multivariate regression analysis suggested that preoperative anemia ( OR=7.922, 95% CI: 1.468-42.757) and biliary sludge and calculus ( OR=1.295, 95% CI: 1.075-4.359) were independent risk factors for postoperative complications; postoperative albumin ( OR=0.055, 95% CI: 0.012-0.244) was a protective factor for postoperative complications, and the differences were statistically significant (all P<0.05). Conclusions:The larger the cyst diameter, the more the intraoperative bleeding, and the higher the risk of operation.Treating anemia before operation, clearing sludge in the hepatic duct during operation, reducing bleeding, and strengthening the monitoring of albumin and hemoglobin during the perioperative period can prevent and reduce early complications after radical resection of CCC in children.

Objective:To explore rehabilitation effect of hyperbaric oxygen chamber treatment on patients with sequelae of brain injury.Methods:From January 2023 to January 2024,a total of 80 patients with sequelae of brain injury were selected from The First Affiliated Hospital of Xinjiang Medical University,and they were randomly divided into control group and observation group,with 40 patients in each group.The control group received conventional treatment and intervention,while the observation group was treated with hyperbaric oxygen chamber on the basis of the conventional treatment and intervention of control group.The levels of serum oxidative stress indicators,neurotrophic factor indicator,as well as neurological function,cognitive function,motor function and quality of life of the two groups were evaluated.Results:After treatment,the levels of serum oxidative stress indicators such as neutrophil gelatinase-related lipocalin(NGAL),matrix metalloproteinase-9(MMP-9),reactive oxygen species(ROS)of observation group were significantly higher than those of control group(t=5.578,11.065,3.706,P<0.05),respectively,and the ubiquitin carboxy-terminal hydrolase L1(UCH-L1)of neurotrophic factor,neuron-specific enolase(NSE)and serum calcium-binding protein(S100-β)of observation group were significantly lower than those of control group,while cerebral extraction rate of oxygen(CERO2)of serum oxidative stress indicators of observation group was significant higher than that of control group,and the differences of them were significantly(t=5.170,4.277,4.130,2.260,P<0.05),respectively.After treatment,the score of National Institutes of Health Stroke Scale(NIHSS)of observation group was lower than that of control group,and the score of Mini-Mental State Examination(MMSE)of observation group was significantly higher than that of control group,and the differences of them were statistical significance(t=15.874,7.820,P<0.05),respectively.In additions,the scores of Loewenstein Occupational Therapy Cognitive Assessment(LOTCA),Simplified Fugl-Meyer Assessment(FMA)upper and lower limbs of observation group were significantly higher than those of control group,and the differences of them between two groups were significant(t=14.051,4.766,8.622,P<0.05),respectively.The scores of physiological,psychological,social and environment factor of World Health Oganization Quality of Life Scale(WHOQOL-BREF)in observation group were higher than those in control group,and the differences of them between two groups were statistically significant(t=19.968,21.979,18.077,19.695,P<0.05).Conclusion:The application of hyperbaric oxygen chamber in patients with sequelae of brain injury is beneficial to adjust the levels of serum oxidative stress indicator and neurotrophic factor indicator,which can improve the deficit of neurological function,and promote the recovery of cognitive and motor functions,and improve the quality of life.

OBJECTIVE: To explore the clinical characteristics and genetic etiology of three children with Menkes disease. METHODS: Three children who had presented at the Children's Medical Center, the Affiliated Hospital of Guangdong Medical University from January 2020 to July 2022 were selected as the study subjects. Clinical data of the children were reviewed. Genomic DNA was extracted from peripheral blood samples of the children, their parents and sister of child 1. Whole exome sequencing (WES) was carried out. Candidate variants were verified by Sanger sequencing, copy number variation sequencing (CNV-seq), and bioinformatic analysis. RESULTS: Child 1 was a 1-year-and-4-month male, and children 2 and 3 were monozygotic twin males aged 1-year-and-10-month. The clinical manifestations of the three children have included developmental delay and seizures. WES showed that child 1 has harbored a c.3294+1G>A variant of the ATP7A gene. Sanger sequencing confirmed that his parents and sister did not carry the same variant, suggesting that it was de novo. Children 2 and 3 had carried a c.77266650_77267178del copy number variation. CNV-seq results showed that their mother has carried the same variant. By searching the HGMD, OMIM and ClinVar databases, the c.3294+1G>A was known to be pathogenic. No carrier frequency has been recorded in the 1000 Genomes, ESP, ExAC and gnomAD databases. Based on the Standards and Guidelines for the Interpretation of Sequence Variants: A Joint Consensus Recommendation of the American College of Medical Genetics and Genomics (ACMG), the ATP7A gene c.3294+1G>A variant was predicted to be pathogenic. The c.77266650_77267178del variant has involved exons 8 to 9 of the ATP7A gene. ClinGen online system score for it was 1.8, which was also considered to be pathogenic. CONCLUSION The c.3294+1G>A and c.77266650_ 77267178del variants of the ATP7A gene probably underlay the Menkes disease in the three children. Above finding has enriched the mutational spectrum of Menkes disease and provided a basis for clinical diagnosis and genetic counseling.
Humans ; Male ; Computational Biology ; Copper-Transporting ATPases/genetics* ; DNA Copy Number Variations ; Exons ; Menkes Kinky Hair Syndrome/genetics* ; Mutation ; Peptide Fragments ; Seizures ; Infant