OBJECTIVE To explore the effect and mechanism of Zhiqiao gancao decoction （ZQGCD） on pathological angiogenesis of degenerative intervertebral disc. METHODS The rats were randomly divided into sham operation group （normal saline）， model group （normal saline）， hypoxia inducible factor-1α （HIF-1α） inhibitor （YC-1） group [2 mg/（kg·d）， tail vein injection]， and ZQGCD low-dose， medium-dose and high-dose groups [3.06， 6.12， 12.24 g/（kg·d）]， with 8 rats in each group. Except for sham operation group， lumbar disc degeneration model of rat was constructed in all other groups. After modeling， they were given relevant medicine once a day， for consecutive 3 weeks. After the last medication， pathological changes and angiogenesis of the intervertebral disc tissue in rats were observed； the levels of inflammatory factors [interleukin-1β （IL-1β）， IL-6， tumor necrosis factor-α （TNF-α）] and the expressions of angiogenesis-related proteins [HIF-1α， vascular endothelial growth factor （VEGF）， VEGF receptor 2 （VEGFR2）， angiotensin 1（Ang 1）， Ang 2] in the com intervertebral disc tissue in rats were all determined. In cell experiment， the primary nucleus pulposus cells were isolated and cultured from rats， and cellular degeneration was induced using 50 ng/mL TNF-α. The cells were divided into blank control group （10% blank control serum）， TNF-α group （10% blank control serum）， YC-1 group （10% blank control serum+0.2 mmol/L YC-1）， and 5%， 10%， 15% drug-containing serum group （5%， 10%， 15% drug-containing serum）. After 24 hours of intervention， the nucleus pulposus cells were co-cultured with HUVEC. The expressions of Collagen Ⅱ， matrix metalloproteinase-3 （MMP-3） in nucleus pulposus cells were detected. HUVEC proliferation， migration and tube forming ability were detected， and the expression levels of the HIF-1α/VEGF/Ang signal axis and angiogenesis- related proteins （add MMP-2， MMP-9） in HUVEC were detected. RESULTS Animal experiments had shown that compared with model group， the positive expression of CD31 in the intervertebral disc tissues of rats in each drug group was down-regulated （P＜ 0.05）， the levels of inflammatory factors and angiogenesis-related proteins were decreased significantly （P＜0.05）， and the pathological changes in the intervertebral disc were alleviated. Cell experiments had shown that compared with TNF-α group， the expression of Collagen Ⅱ in nucleus pulposus cells of all drug groups was significantly up-regulated （P＜0.05）， and the expression of MMP-3 was significantly down-regulated （P＜0.05）； the proliferation， migration and tubulogenesis of HUVEC were significantly weakened （P＜0.05）. The mRNA and protein expressions of HIF-1α， VEGF， Ang 2 as well as the expression of angiogenesis-related proteins （except for the expression of Ang 2 mRNA and HIF-1α， VEGFR2， Ang 2 protein in 5% drug- containing serum group） were significantly down-regulated （P＜0.05）. CONCLUSIONS ZQGCD may inhibit the HIF-1α/VEGF/ Ang signal axis to weaken the angiogenic ability of vascular endothelial cells， improve pathological angiogenesis in the intervertebral disc， and delay the degeneration of the intervertebral disc.

Objective:This study aims to investigate the therapeutic efficacy of staged endoscopic surgery for Shin-Ⅲ stage external auditory canal cholesteatoma. Methods:A retrospective analysis was conducted on the clinical data of 25 patients diagnosed with Shin-Ⅲ cholesteatoma of the external auditory canal, who were admitted to the Otology Center of the First People's Hospital of Foshan City from May 2020 to October 2024. All patients initially underwent endoscopic cholesteatoma removal. Based on the outcomes of the first-stage postoperative follow-up, patients were categorized into two groups: the repair type and the non-repair type. The non-repair type was further subdivided into simple and complex types. Of the total cases, 10 patients were of the repair type, with 9 requiring no further surgical intervention. The non-repair type comprised 15 patients, of which 8 were classified as simple type and underwent either tympanoplasty type Ⅰ or external auditory canal wall reconstruction during the second stage. The remaining 7 patients, identified as complex type, received open mastoidectomy or tympanotomy in the second stage, with or without ossicular chain reconstruction. Results:All patients were monitored for a minimum of six months postoperatively. The incidence of dry ear was observed in 22 patients, corresponding to a dry ear rate of 88.0%. Four cases experienced primary complications. Conclusion:Endoscopic phased operation for managing Shin-Ⅲ stage cholesteatoma of the external auditory canal can ensure that the surgical options match the severity of the lesions, reducing unnecessary surgical trauma and achieving good efficacy.
Humans ; Retrospective Studies ; Ear Canal/surgery* ; Endoscopy/methods* ; Cholesteatoma/surgery* ; Male ; Tympanoplasty ; Female ; Treatment Outcome ; Adult ; Middle Aged ; Cholesteatoma, Middle Ear/surgery* ; Mastoidectomy

ObjectiveTo investigate the possible mechanism by which Zhiqiao Gancao Decoction （枳壳甘草汤， ZGD） delays intervertebral disc degeneration （IDD） based on the Hippo-yes-associated protein （YAP）/transcriptional co-activator with PDZ-binding motif （TAZ） signaling pathway. MethodsA total of 50 SD rats were randomly divided into sham surgery group， model group， low-dose ZGD group， high-dose ZGD group， and high-dose ZGD + inhibitor group， with 10 rats in each group. In the sham surgery group， the rats were pierced in the skin and muscle at the Co6/7/8 segments of the tail with a 21G needle （depth approximately 2 mm） without damaging the intervertebral disc. In the other groups， rats were injected with a 21G needle at the Co6/7/8 segments of the tail to establish an IDD model by piercing the tail intervertebral disc 5 mm. One week after modeling， rats in the low-dose and high-dose ZGD groups were given 6.24 and 12.24 g/（kg·d） of the decoction via gastric gavage， respectively. The high-dose ZGD + inhibitor group was given 12.24 g/（kg·d） of the decoction and an intraperitoneal injection of YAP/TAZ inhibitor Verteporfin 10 mg/kg. The sham surgery and model groups were given 5 ml/（kg·d） of normal saline via gavage. The gavage was given once a day， and the intraperitoneal injection was given every other day. After 4 weeks of continuous intervention， the pathological changes of the tail intervertebral discs were observed using HE staining， Oil Red O-Green staining， and Toluidine Blue staining. Immunohistochemistry was used to detect the expression of aggrecan and MMP3 in the nucleus pulposus. TUNEL fluorescence staining was performed to detect apoptosis in the nucleus pulposus， and the apoptosis rate was calculated. Western blot was used to detect the Hippo-YAP/TAZ signaling pathway， including YAP， phosphorylated YAP （p-YAP）， phosphorylated MST1/2 （p-MST1/2）， phosphorylated TAZ （p-TAZ） and apoptosis-related proteins， such as Cleaved Caspase 3， P53， Bcl-2 and Bax. ResultsCompared with sham surgery group， the rats in the model group showed significant degenerative changes in the intervertebral disc. The levels of aggrecan， Bcl-2， and YAP proteins in the nucleus pulposus decreased， while the levels of p-MST1/2， p-YAP， p-TAZ， P53， Bax， Cleaved Caspase 3， MMP3 proteins， and the apoptosis rate increased （P < 0.01）. Compared with the model group， the drug intervention groups showed partial recovery in intervertebral disc degeneration. The levels of aggrecan， Bcl-2， and YAP proteins increased， while the levels of p-MST1/2， p-YAP， p-TAZ， P53， Bax， Cleaved Caspase 3， MMP3 proteins， and the apoptosis rate decreased （P＜0.05 or P＜0.01）. The high-dose ZGD group showed more significant recovery in intervertebral disc degeneration compared to the low-dose ZGD group， with a decrease in the levels of p-MST1/2， p-YAP， p-TAZ， P53， Bax， Cleaved Caspase 3， MMP3 proteins， and apoptosis rate， and an increase in the levels of aggrecan， Bcl-2， and YAP proteins （P＜0.05 or P＜0.01）. Compared with the high-dose ZGD group， the high-dose ZGD + inhibitor group showed a reduced recovery in intervertebral disc degeneration， with an increase in the levels of p-MST1/2， p-YAP， p-TAZ， P53， Bax， Cleaved Caspase 3， MMP3 proteins， and apoptosis rate， and a decrease in the levels of aggrecan， Bcl-2， and YAP proteins （P＜0.05 or P＜0.01）. ConclusionZGD may delay intervertebral disc degeneration by inhibiting the phosphorylation of YAP in the nucleus pulposus， maintaining the function of the Hippo-YAP/TAZ signaling pathway， and reducing apoptosis of nucleus pulposus cells.

Objective:To analyze the different expression of neutrophil's protein from the patients with active pulmonary tuber-culosis(TB)and healthy controls(HC)from the perspective of proteomics,and to explore the diagnosis of the secreted proteins related to neutrophil activation in serum.Methods:Neutrophil protein in peripheral blood of 15 patients with TB and 15 HC was identified by label free quantitative proteomics,which were further validated in by ELISA and nephelometry.Results:A total of 358 differentially expressed proteins were identified in our study,54 differentially expressed proteins related to neutrophil activation or degranulation were focused,and seven proteins AHSG,AACT,C3,AAT,AAG,HP and A1BG were validated in 40 TB patients and 40 HC,and the results confirmed that the following proteins were highly expressed in the serum of the TB group:AACT,C3,AAT,AAG,HP and A1BG,while AHSG was lowly expressed,and only the validation results of AHSG were consistent with the trend of proteomics results.In addition,we further investigated the expression level of AHSG in the diagnosis of 24 patients with active pulmonary TB,24 patients with lung cancer,24 HC,and 16 patients with latent tuberculosis infection(LTBI),and found the expression of AHSG was down-regu-lated in TB group compared with the other groups.The sensitivity and specificity of AHSG in differentiating TB patients from HC patients,lung cancer patients,LTBI patients and non-TB patients in the test set were 83.33%,79.17%;78.00%,91.67%;75.00%,87.50%;75.00%,85.94%.Conclusion:Proteomic profiles of neutrophils from TB and HC are obtained from a proteomic perspective,and further validation shows that AHSG has a good differential diagnostic ability for HC,lung cancer and LTBI.

Objective To assess the efficacy of clear lens extraction combined with extended depth of focus(EDOF)intraocular lens(IOL)implantation in enhancing visual quality and concurrently opening peripheral angle structures in patients diagnosed with primary angle closure suspect(PACS)and presbyopia.Methods A total of 80 patients with PACS,presbyopia,and transparent lenses were enrolled and divided into two groups based on the type of implanted lens:a control group(n=46)and an experimental group(n=34).The control group underwent clear lens extraction followed by monofocal intraocular lens implantation,while the experimental group underwent clear lens extraction followed by extended depth of focus(EDOF)intraocular lens implantation.Preoperative and postoperative evaluations at 3 months included visual acuity,intraocular pressure,anterior segment structure,defocus curve,and Catquest 9SF-CN questionnaire scores.Adverse events were recorded to assess safety.Results Regarding the angular structure,after 3 months of treatment,results indicated significant increases in central ante-rior chamber depth,angle opening distance at 750 μ m,angle recess area at 750 μ m,trabecular-iris space area at 750 μm,and trabecular-iris angle at 750 μm in both groups compared to preoperative values(all P<0.01).However,no significant differences were observed between the two groups(all P>0.05).For the defocusing curve,visual acuity in the experimental group improved from+1.0 D to-4.0 D after 3 months of treatment(all P<0.05),while the control group showed improvements at+0.5 D,-0.5 D,-1.0 D,-2.0 D,-3.0 D,-3.5 D,and-4.0 D(all P<0.05).Postoperatively at 3 months,the experimental group exhibited superior visual acuity to the control group at-1.0 D to-4.0 D(all P<0.05).According to the Catquest 9SF-CN scale,scores in both groups significantly im-proved after 3 months of treatment(P<0.01),with the experimental group scoring higher than the control group(P<0.01).During the study,transient intraocular pressure elevation occurred in 7 patients(3 in the control group and 4 in the experimental group),but no serious adverse reactions were reported in either group.Conclusions In patients with pseudoexfoliation glaucoma combined with presbyopia,significant improvements in angle structures were observed following clear lens extraction.The implantation of an EDOF IOL notably enhanced postoperative visual quality compared to monofocal IOLs.

Objective:To investigate clinicopathological and molecular genetic characteristics of CD117-positive eosinophilic renal cell tumors (ERCTs) with unusual morphological and immunophenotypic features.Methods:Formalin-fixed, paraffin-embedded tissues from 10 cases (9 cases from Xiangya Hospital, Central South University and 1 case from Bishan Hospital of Chongqing Medical University) of diagnostically challenging CD117-positive ERCTs between January 2017 and October 2024 were collected. Histological reviews were performed on HE-stained sections, followed by immunostaining and whole-exome sequencing (WES).Results:The 10 patients were composed of 4 males and 6 females, with ages ranging from 29 to 57 years, median 49.5 (36.8, 51.8) years. The sizes of tumors ranged from 2.5 to 6.0 cm, median 4.8(2.9,5.2) cm. All 10 ERCTs were composed of variably eosinophilic cells and characterized by prominent morphological features including exclusively eosinophilic (2 cases), focal chromophobe-like (3 cases), prominent nested (2 cases), prominent flocculent cytoplasm (1 case), a collision of renal oncocytoma (RO)/chromophobe renal cell carcinoma (ChRCC) (1 case), and diffusely degenerative atypia (1 case). Immunohistochemically, a subset of 10 tumors variably expressed CK7 (7/10) and vimentin (3/10), while they were all positive for CD117 (10/10), PAX8 (10/10), SDHB (10/10), and FH (10/10) and negative for CAⅨ (10/10) and 2SC (10/10). The Ki-67 proliferation index ranged from 1% to 5%. WES identified a GNAS mutation in one case of the RO/ChRCC collision tumor, while no characteristic mutations of other renal cell tumor types were detected in the remaining 9 cases. The analysis of copy number variations revealed complex karyotypic alterations in 4 tumors, harboring various gain of chromosomes 4, 5, 7, 12, 13, 15, 16, 18, and 22, with 3 cases showing variable loss of chromosomes 1, 2, 6, 10, 13, and 17. These 4 cases were molecularly classified as eosinophilic ChRCC. The remaining 6 cases, including 2 cases with a normal diploid karyotype and 4 cases with slight karyotypic alterations, were molecularly classified as 5 ROs and 1 RO-dominant RO/ChRCC collision tumor. Finally, the original diagnosis was retained in 4 cases and revised in 6 cases.Conclusions:CD117-positive ERCTs with uncertain classification may exhibit various morphological overlaps, non-classic histological features, and aberrant immunophenotypes. Combined immunostaining of CK7, CD117, vimentin, SDHB, FH, and 2SC can greatly help discriminate among these tumors and their mimics. When the diagnosis is challenging based only on morphology and immunohistochemistry, molecular genetic tests may be useful.

Objective:To analyze the different expression of neutrophil's protein from the patients with active pulmonary tuber-culosis(TB)and healthy controls(HC)from the perspective of proteomics,and to explore the diagnosis of the secreted proteins related to neutrophil activation in serum.Methods:Neutrophil protein in peripheral blood of 15 patients with TB and 15 HC was identified by label free quantitative proteomics,which were further validated in by ELISA and nephelometry.Results:A total of 358 differentially expressed proteins were identified in our study,54 differentially expressed proteins related to neutrophil activation or degranulation were focused,and seven proteins AHSG,AACT,C3,AAT,AAG,HP and A1BG were validated in 40 TB patients and 40 HC,and the results confirmed that the following proteins were highly expressed in the serum of the TB group:AACT,C3,AAT,AAG,HP and A1BG,while AHSG was lowly expressed,and only the validation results of AHSG were consistent with the trend of proteomics results.In addition,we further investigated the expression level of AHSG in the diagnosis of 24 patients with active pulmonary TB,24 patients with lung cancer,24 HC,and 16 patients with latent tuberculosis infection(LTBI),and found the expression of AHSG was down-regu-lated in TB group compared with the other groups.The sensitivity and specificity of AHSG in differentiating TB patients from HC patients,lung cancer patients,LTBI patients and non-TB patients in the test set were 83.33%,79.17%;78.00%,91.67%;75.00%,87.50%;75.00%,85.94%.Conclusion:Proteomic profiles of neutrophils from TB and HC are obtained from a proteomic perspective,and further validation shows that AHSG has a good differential diagnostic ability for HC,lung cancer and LTBI.

Objective To assess the efficacy of clear lens extraction combined with extended depth of focus(EDOF)intraocular lens(IOL)implantation in enhancing visual quality and concurrently opening peripheral angle structures in patients diagnosed with primary angle closure suspect(PACS)and presbyopia.Methods A total of 80 patients with PACS,presbyopia,and transparent lenses were enrolled and divided into two groups based on the type of implanted lens:a control group(n=46)and an experimental group(n=34).The control group underwent clear lens extraction followed by monofocal intraocular lens implantation,while the experimental group underwent clear lens extraction followed by extended depth of focus(EDOF)intraocular lens implantation.Preoperative and postoperative evaluations at 3 months included visual acuity,intraocular pressure,anterior segment structure,defocus curve,and Catquest 9SF-CN questionnaire scores.Adverse events were recorded to assess safety.Results Regarding the angular structure,after 3 months of treatment,results indicated significant increases in central ante-rior chamber depth,angle opening distance at 750 μ m,angle recess area at 750 μ m,trabecular-iris space area at 750 μm,and trabecular-iris angle at 750 μm in both groups compared to preoperative values(all P<0.01).However,no significant differences were observed between the two groups(all P>0.05).For the defocusing curve,visual acuity in the experimental group improved from+1.0 D to-4.0 D after 3 months of treatment(all P<0.05),while the control group showed improvements at+0.5 D,-0.5 D,-1.0 D,-2.0 D,-3.0 D,-3.5 D,and-4.0 D(all P<0.05).Postoperatively at 3 months,the experimental group exhibited superior visual acuity to the control group at-1.0 D to-4.0 D(all P<0.05).According to the Catquest 9SF-CN scale,scores in both groups significantly im-proved after 3 months of treatment(P<0.01),with the experimental group scoring higher than the control group(P<0.01).During the study,transient intraocular pressure elevation occurred in 7 patients(3 in the control group and 4 in the experimental group),but no serious adverse reactions were reported in either group.Conclusions In patients with pseudoexfoliation glaucoma combined with presbyopia,significant improvements in angle structures were observed following clear lens extraction.The implantation of an EDOF IOL notably enhanced postoperative visual quality compared to monofocal IOLs.

Objective:To investigate clinicopathological and molecular genetic characteristics of CD117-positive eosinophilic renal cell tumors (ERCTs) with unusual morphological and immunophenotypic features.Methods:Formalin-fixed, paraffin-embedded tissues from 10 cases (9 cases from Xiangya Hospital, Central South University and 1 case from Bishan Hospital of Chongqing Medical University) of diagnostically challenging CD117-positive ERCTs between January 2017 and October 2024 were collected. Histological reviews were performed on HE-stained sections, followed by immunostaining and whole-exome sequencing (WES).Results:The 10 patients were composed of 4 males and 6 females, with ages ranging from 29 to 57 years, median 49.5 (36.8, 51.8) years. The sizes of tumors ranged from 2.5 to 6.0 cm, median 4.8(2.9,5.2) cm. All 10 ERCTs were composed of variably eosinophilic cells and characterized by prominent morphological features including exclusively eosinophilic (2 cases), focal chromophobe-like (3 cases), prominent nested (2 cases), prominent flocculent cytoplasm (1 case), a collision of renal oncocytoma (RO)/chromophobe renal cell carcinoma (ChRCC) (1 case), and diffusely degenerative atypia (1 case). Immunohistochemically, a subset of 10 tumors variably expressed CK7 (7/10) and vimentin (3/10), while they were all positive for CD117 (10/10), PAX8 (10/10), SDHB (10/10), and FH (10/10) and negative for CAⅨ (10/10) and 2SC (10/10). The Ki-67 proliferation index ranged from 1% to 5%. WES identified a GNAS mutation in one case of the RO/ChRCC collision tumor, while no characteristic mutations of other renal cell tumor types were detected in the remaining 9 cases. The analysis of copy number variations revealed complex karyotypic alterations in 4 tumors, harboring various gain of chromosomes 4, 5, 7, 12, 13, 15, 16, 18, and 22, with 3 cases showing variable loss of chromosomes 1, 2, 6, 10, 13, and 17. These 4 cases were molecularly classified as eosinophilic ChRCC. The remaining 6 cases, including 2 cases with a normal diploid karyotype and 4 cases with slight karyotypic alterations, were molecularly classified as 5 ROs and 1 RO-dominant RO/ChRCC collision tumor. Finally, the original diagnosis was retained in 4 cases and revised in 6 cases.Conclusions:CD117-positive ERCTs with uncertain classification may exhibit various morphological overlaps, non-classic histological features, and aberrant immunophenotypes. Combined immunostaining of CK7, CD117, vimentin, SDHB, FH, and 2SC can greatly help discriminate among these tumors and their mimics. When the diagnosis is challenging based only on morphology and immunohistochemistry, molecular genetic tests may be useful.

Objective To employ a mouse model of ABI3BP gene deletion for the detection of postnatal changes in body weight and glucose metabolism and establish a different method of creating a mouse model of low birth weight.Methods Heterozygote mice were mated to produce ABI3BP gene knockout homozygote(ABI3BP-/-)mice,heterozygote(ABI3BP+/)mice,and wild-type(WT)mice.Adult mice from all three groups were evaluated for glucose metabolism markers,including the fasting blood glucose level,glucose tolerance,and insulin tolerance.Additionally,body weight was measured at various postnatal time periods,and the weight ratio of critical organs in adulthood was calculated.Results The gene sequencing result of the polymerase chain reaction product of ABI3BP-/-mice showed that frameshift mutations occurred in the knockout region,with quantitative reverse-transcription polymerase chain reaction analysis demonstrating significantly reduced ABI3BP expression in ABI3BP-/-mice compared with that in WT mice.Notably,the birth weight of ABI3BP-/-mice(1.25±0.08 g)was markedly lower than that of WT mice(1.34±0.12 g)(P＜0.05).Conversely,the weight of adult(120 d)ABI3BP-/mice(27.70±1.93 g)was significantly higher than that of WT mice(23.64±1.34 g)(P＜0.01).The ratios of key organ weights to body weight were not significantly different between the groups(P＞0.05).Fasting blood glucose and insulin tolerance tests showed no significant variations between the groups.However,glucose tolerance tests indicated that ABI3BP-/-mice had lower blood glucose levels(15.68±7.04 mmol/L)than WT mice(23.01±5.75 mmol/L).Conclusions Deletion of the ABI3BP gene result in mice with low birth weight,poor growth recuperation,and inadequate glucose tolerance in adulthood,similar to the clinical growth traits of low-birth-weight human neonates.Therefore,this mouse model is a promising choice for the study of low birth weight.