AIM To establish a UPLC-MS/MS method for the simultaneouscontent determination of notoginsenosides R1,Fc,Fe,ginsenosides Re,Rg1,Rf,F3,Rg2,Ra2,Rb1,Ro,Rc,F1,Ra1,Rb2,Rb3,Rd,F2,Rg5,chikusetsusaponin Ⅳ a,20(S)-ginsenoside Rg3 and 20(R)-ginsenoside Rg3 in Dengzhan Shengmai Capsules.METHODS The analysis was performed on a 35 ℃ thermostatic Accucore Phenyl Hexyl column(2.1 mm×100 mm,2.6 μm),with the mobile phase comprising of 0.1％formic acid-acetonitrile flowing at 0.4 mL/min in a gradient elution manner,and heated electrospray ionization source was adopted in negative ion scanning with parallel reaction monitoring mode.Subsequently,cluster analysis,principal component analysis and partial least squares discriminant analysis were adopted.RESULTS Twenty-two saponins showed good linear relationships within their own ranges(r ≥ 0.996 4),whose average recoveries were 98.5％-101.6％with the RSDs of 1.3％-5.2％.Ten batches of samples were clustered into four types,three principal components demonstrated the accumulative variance contribution rate of 90.265％,ginsenosides Rb1,Rg2,Rb2,Rd,Rg1,Ro,Rf,Re,Rg5,Rc were taken as potential quality markers.CONCLUSION This simple,efficient,accurate and sensitive method can be used for the quality control of Dengzhan Shengmai Capsules.

Objective To observe the value of shear wave elastography(SWE)for evaluating therapeutic effect of ultrasound-guided drug injection for muscle injury.Methods Eighty patients with unilateral muscle injury were retrospectively included,including 40 cases underwent ultrasound-guided drug injection(group A)and 40 cases underwent electromagnetic wave physiotherapy plus external application of Yunnan Baiyao Gao(group B).Pain intensity was assessed using visual analogue scale(VAS)before treatment and 3 weeks after the final treatment,while the Young modulus(E)value of the injured muscle was measured before treatment and 1,2 and 3 weeks after final treatment,and the improvements of VAS scores and E values were compared between groups.Results The total effective rate in group A(35/40,87.50%)was higher than that in group B(21/40,52.50%;P＜0.05).Before treatment,no significant difference of VAS score was found between group A(8.07±0.83)and group B(7.88±0.85)(P＞0.05).After treatment,VAS scores decreased in both groups(both P＜0.05),which in group A(2.30±1.07)was more obviously than that in group B(4.80±0.82)(P＜0.05).After treatment,E values of injury muscles increased significantly in both groups(P＜0.05),while group A had a greater increase in overall magnitude and overall rate than group B(P＜0.05).Conclusion Ultrasound-guided drug injection therapy had significant therapeutic effect for muscle injuries,which could be dynamically monitored with SWE.

Objective:To explore the role of nuclear receptor-binding SET-domain protein 1(NSD1)in the pathogenesis of nonobstructive azoospermia(NOA)by regulating the expressions of relevant genes.Methods:We detected the expression of NSD1 in the testis tissue of 7 male patients with obstructive azoospermia(OA)and 18 with NOA by qPCR and immunofluorescence assay,and determined the modification level of H3K36me2 in the testes of two groups of patients by immunofluorescence staining,Western blot and immunoprecipitation(IP).We examined the difference in the enrichment of H3K36me2 in the testis tissue by chromatin IP-based sequencing(ChIP-Seq),analyzed the genomic distribution and target genes using bioinformatics,and verified the expression levels of the target genes in the testes of the two groups of patients by qPCR.Results:Compared with the patients with OA,those with NOA showed dramatically decreased mRNA and protein expressions of NSD1(P=0.000 8).The binding of NSD1 to H3K36me2 was observed in the testis tissue of both the two groups of patients,while the modification level of H3K36me2 was evidently reduced in the NOA males.H3K36me2 was distributed mainly in the intergenic region in the testes of the two groups of patients,but the enrich-ment of H3K36me2 was obviously decreased in the NOA group.The differentially H3K36me2-enriched genes were involved in various biological processes,including tissue development,and cell morphogenesis.Results of ChIP-Seq and qPCR showed significantly down-regulated expressions of the target genes KIT,SPO11 and ACRV1 in the testis tissue of the NOA males compared with those in the OA patients(P＜0.01).Conclusion:The levels of NSD1 and H3K36me2 are decreased in testis tissue of the NOA patient,H3K36me2 is highly enriched in the spermatogenesis-related key genes KIT,SPO11 and ACRV1,and the down-regulated expression of NSD1 impairs spermatogenesis.

Objective:To explore the relationship between retinol-binding protein (RBP) levels and disease severity in patients with transthyretin cardiac amyloidosis (ATTR-CA), as well as its impact on therapeutic response to tafamidis.Methods:This retrospective study utilized data from the China National Rare Disease Registry System and included ATTR-CA patients treated with tafamidis between January 2018 and September 2022. Patients were stratified into two groups based on baseline RBP levels: the normal RBP group (≥36 mg/L) and the reduced RBP group (<36 mg/L). Baseline characteristics and clinical data after one year of treatment were collected and compared between the groups. Within the reduced RBP group, patients were further subclassified by changes in RBP levels after treatment (ΔRBP=post-treatment RBP-baseline RBP) into ΔRBP>0 and ΔRBP<0 subgroups. Worsening of global longitudinal strain (GLS) after treatment was defined as the primary outcome, logistic regression analysis was used to identify risk factors influencing therapeutic response to tafamidis in ATTR-CA patients.Results:A total of 52 ATTR-CA patients were included (aged (58.5±12.0) years, 46 males (88%)). Among 39 patients who completed one-year tafamidis treatment, no statistically significant difference was observed in RBP levels post-treatment versus baseline ((27.0±14.3) mg/L vs. (25.9±15.4) mg/L, P=0.261). Compared to the normal RBP group, the reduced RBP group had significantly higher estimated glomerular filtration rate-adjusted N-terminal pro-B-type natriuretic peptide levels (2 316.0 (1 161.5, 6 027.8) ng/L vs. 806.2 (349.5, 1 735.8) ng/L), higher left ventricular mass index ((164.4±46.5) g/m2 vs. (123.9±31.8) g/m2), and lower left ventricular ejection fraction ((50.8±11.3)% vs. (58.8±6.2)%) (all P<0.05). Among 31 patients in the reduced RBP group who completed one-year tafamidis treatment, 23 were classified as ΔRBP>0 and 8 as ΔRBP<0. The ΔRBP<0 group exhibited greater GLS worsening than the ΔRBP>0 group (0.7 (-0.1, 1.4)% vs. -0.4 (-1.4, 0.2)%, P=0.027). Multivariate logistic regression analysis revealed that ΔRBP<0 was an independent risk factor for GLS worsening ( OR=8.584, 95%CI 1.186-62.150, P=0.033) in ATTR-CA patients. Conclusion:ATTR-CA patients with reduced RBP levels exhibit more severe left ventricular structural and functional impairment compared to those with normal RBP levels. Decline in RBP during treatment (ΔRBP<0) is associated with poorer response to tafamidis treatment. Monitoring RBP dynamics may assist clinicians in assessing disease severity and therapeutic response in ATTR-CA patients.

Sacroiliac complex injuries are commonly seen in high-energy pelvic fractures. The injuries make a big difference in treatment patterns due to the diverse injury types, posing considerable challenges in formulating optimal treatment strategies, and hence are persistent clinical difficulties in orthopedic trauma. The clinical management of sacroiliac complex injuries presents several key challenges such as a non-negligible rate of missed diagnoses in associated vascular and visceral injuries, absence of standardized protocols for surgical approaches and reduction-fixation strategies across different injury patterns, and ongoing controversies regarding surgical indications and optimal timing for patients combined with concomitant lumbosacral plexus injuries. Currently, no systematic clinical guidelines are available for the diagnosis and treatment of sacroiliac complex injuries both domestically and internationally. To this end, the Pelvic and Acetabular Surgery Group, Orthopedic Branch, China International Exchange and Promotive Association for Medical and Health Care and Orthopedic Physician Branch, Chinese Medical Doctor Association organized a panel of domestic experts in the field to develop the Clinical guideline for the diagnosis and treatment of sacroiliac complex injuries ( version 2025), based on evidence-based medicine and adhering to the principles of scientific rigor, clinical applicability, and innovation. These guidelines provided 11 recommendations covering diagnosis, therapeutic principles and techniques, management protocols for lumbosacral plexus injuries, outcome evaluation, and postoperative rehabilitation pathways, etc., aiming to standardize the clinical management of sacroiliac complex injuries.

Objective:To determine the prevalence of GJB2 gene c. 109G>A (p.V37I) variant among infants with congenital hearing loss and analyze the initial audiological characteristics of children harboring the variant, compare the audiometric difference among individuals with various genotypes, and explore genetic and audiological manifestations of the affected families. Methods:One hundred twenty six infants diagnosed with congenital hearing loss at the Neonate Screening Center of Ningbo City from June 2021 to December 2024 were selected as the study subjects. The neonates, in addition with members from 16 of their families, had undergone genetic screening for variants of 208 hotspot sites within 24 deafness-associated genes. For cases identified with monoallelic variants and concurrent hearing loss, the full GJB2 gene was sequenced. Meanwhile, a retrospective analysis was carried out on 23 children whom were confirmed to have hearing loss and the c. 109G>A variant by whole exome sequencing from March 2022 to December 2024. And 102 children who were excluded to have hearing loss and pathogenic variants by whole exome sequencing were selected as normal controls. Audiological features of individuals harboring the c. 109G>A variant were compared. This study has been approved by the Medical Ethics Committee of The Affiliated Women and Children′s Hospital of Ningbo University (Ethics No.: EC2023-009). Results:For the 126 infants with congenital hearing loss, prospective screening has identified 58 (46.03%) to harbor the c. 109G>A variant. These included 38 homozygotes and 16 compound heterozygotes. Retrospective review of the 23 c. 109G>A positive children has identified 15 as homozygotes and 8 as compound heterozygotes. Genetic testing of the 16 pedigrees has identified 7 homozygotes and 1 compound heterozygote. For the homozygotes combined ( n=53), 96.2% exhibited bilateral symmetric hearing loss, with 78.3% showing high-frequency sloping patterns, and 98.1% having a hearing threshold ranging from 20 to 65 dB. For the compound heterozygotes combined ( n=24), 95.8% showed symmetric loss, with 59.4% having high-frequency sloping, and 97.9% had a hearing threshold ranging from 20 to 65 dB. Both groups showed significantly elevated ABR/PTA thresholds compared with the normal controls ( P=0.000). The compound heterozygous group had higher ABR thresholds (43.3 ± 15.0 dB nHL) compared with the homozygous group (39.1±12.0 dB nHL, P=0.005). Conclusion:Infants harboring the GJB2 c. 109G>A variant primarily manifest as mild-to-moderate, symmetric, high-frequency sloping hearing loss. Nearly one-third of affected children have thresholds between 20 to 35 dB nHL, suggesting that ABR > 35 dB nHL alone may underestimate the hearing impairment in this population. Compared with homozygotes, compound heterozygotes with the the GJB2 c. 109G>A variant can confer a more severe hearing loss.

Objective:To investigate the clinical and pathological characteristics, prognostic factors, and treatment outcomes of bladder adenocarcinoma.Methods:The data of 177 bladder adenocarcinoma patients treated at Renji Hospital, Shanghai Jiaotong University School of Medicine from January 2003 to December 2023, and 2 687 bladder adenocarcinoma patients from the SEER database (2000—2021) were reviewed retrospectively. The clinicopathological and prognostic characteristics were compared between the two cohorts. Patients with urachal adenocarcinoma or primary bladder adenocarcinoma were included, while metastatic bladder adenocarcinoma from other sites and urothelial carcinoma with glandular components were excluded. The Chi-square test was used for comparison of categorical data, and propensity score matching (1∶1) was applied to match baseline data between the Renji and SEER cohorts. Kaplan-Meier survival curves were generated, and log-rank tests were used for comparisons. Univariate and multivariate Cox regression analyses were performed using the survival R package, with P-values calculated via Wald tests. Results:The proportion of localized bladder adenocarcinoma was significantly higher in the Renji cohort than in the SEER cohort [61.0% (108/177) vs. 19.4% (521/2 687), P<0.001], and mucinous adenocarcinoma was more common in Renji cohort [33.3% (59/177) vs. 22.6% (607/2 687), P<0.001]. After matching for baseline factors, including SEER stage and pathological grade, survival analysis revealed that the Renji cohort patients had slightly better survival compared to the SEER cohort [median survival: 55.4 (24.1, 196.2) months vs. 39.2 (13.6, 137.4)months, P=0.033]. Multivariate Cox analysis identified SEER stage [Renji cohort: HR=3.83 (95% CI 1.62-9.07), P=0.002; SEER cohort: HR=3.67 (95% CI 3.13-4.31), P<0.001] and pathological grade [Renji cohort: HR = 2.76 (95% CI 1.54-4.95), P=0.001; SEER cohort: HR=1.46 (95% CI 1.29-1.65), P<0.001] as independent prognostic factors. In the Renji cohort, no significant differences were observed in the median progression-free survival [40.1 (19.5, 91.6) months vs. 40.9 (12.8, not reached)months, P=0.976] and overall survival [79.3 (37.1, 195.8) months vs. 53.9 (16.4, 129.5)months, P=0.374] between patients receiving adjuvant chemotherapy and those not receiving it. However, among patients with lymph node-positive bladder adenocarcinoma, adjuvant chemotherapy significantly improved both progression-free survival [40.1 (38.2, 75.4) months vs. 12.2 (3.1, 12.2)months, P=0.004] and overall survival [68.2 (46.2, 84.5)months vs. 28.1 (4.3, 28.3)months, P=0.006]. Conclusions:Bladder adenocarcinoma is rare and associated with poor prognosis. Compared to the SEER cohort, Renji cohort patients had more localized disease, with no significant differences in other features. SEER stage and pathological grade were independent prognostic factors in both cohorts. Lymph node-positive bladder adenocarcinoma patients in the Renji cohort benefited significantly from adjuvant chemotherapy.

Osteoporosis is a common bone disease， whose incidence is still on the rise， posing great challenges to patients and society. This review mainly studies the pathogenesis of osteoporosis from the aspects of oxidative stress， inflammatory response， and glucolipotoxicity-induced injury and clarifies the efficacy and mechanism of some active ingredients of traditional Chinese medicine against osteoporosis through the integration of in vitro and in vivo experiments. The experimental results suggest that some active ingredients can improve bone resorption markers and maintain bone homeostasis by modulating inflammation， oxidative stress， etc. These active ingredients regulate osteoporosis through the receptor activator of nuclear transcription factor-κB （NF-κB） ligand （RANKL） pathway， osteoprotegerin （OPG） pathway， Wnt/β-catenin pathway， NF-κB pathway， mitogen-activated protein kinase （MAPK） pathway， adenosine monophosphate （AMP）-activated protein kinase （AMPK）/mammalian target of rapamycin （mTOR） pathway， and oxidative stress pathway. This review provides ideas for the progress of the prevention and treatment of osteoporosis with the active ingredients of traditional Chinese medicine， aiming to provide new potential lead compounds and reference for the development of innovative drugs and clinical therapy for the treatment of osteoporosis.

OBJECTIVE: To determine the prevalence of GJB2 gene c.109G>A (p.V37I) variant among infants with congenital hearing loss and analyze the initial audiological characteristics of children harboring the variant, compare the audiometric difference among individuals with various genotypes, and explore genetic and audiological manifestations of the affected families. METHODS: One hundred twenty six infants diagnosed with congenital hearing loss at the Neonate Screening Center of Ningbo City from June 2021 to December 2024 were selected as the study subjects. The neonates, in addition with members from 16 of their families, had undergone genetic screening for variants of 208 hotspot sites within 24 deafness-associated genes. For cases identified with monoallelic variants and concurrent hearing loss, the full GJB2 gene was sequenced. Meanwhile, a retrospective analysis was carried out on 23 children whom were confirmed to have hearing loss and the c.109G>A variant by whole exome sequencing from March 2022 to December 2024. And 102 children who were excluded to have hearing loss and pathogenic variants by whole exome sequencing were selected as normal controls. Audiological features of individuals harboring the c.109G>A variant were compared. This study has been approved by the Medical Ethics Committee of The Affiliated Women and Children's Hospital of Ningbo University (Ethics No.: EC2023-009). RESULTS: For the 126 infants with congenital hearing loss, prospective screening has identified 58 (46.03%) to harbor the c.109G>A variant. These included 38 homozygotes and 16 compound heterozygotes. Retrospective review of the 23 c.109G>A positive children has identified 15 as homozygotes and 8 as compound heterozygotes. Genetic testing of the 16 pedigrees has identified 7 homozygotes and 1 compound heterozygote. For the homozygotes combined (n = 53), 96.2% exhibited bilateral symmetric hearing loss, with 78.3% showing high-frequency sloping patterns, and 98.1% having a hearing threshold ranging from 20 to 65 dB. For the compound heterozygotes combined (n = 24), 95.8% showed symmetric loss, with 59.4% having high-frequency sloping, and 97.9% had a hearing threshold ranging from 20 to 65 dB. Both groups showed significantly elevated ABR/PTA thresholds compared with the normal controls (P = 0.000). The compound heterozygous group had higher ABR thresholds (43.3 ± 15.0 dB nHL) compared with the homozygous group (39.1 ± 12.0 dB nHL, P = 0.005). CONCLUSION Infants harboring the GJB2 c.109G>A variant primarily manifest as mild-to-moderate, symmetric, high-frequency sloping hearing loss. Nearly one-third of affected children have thresholds between 20 to 35 dB nHL, suggesting that ABR > 35 dB nHL alone may underestimate the hearing impairment in this population. Compared with homozygotes, compound heterozygotes with the the GJB2 c.109G>A variant can confer a more severe hearing loss.
Humans ; Connexin 26/genetics* ; Female ; Male ; Infant, Newborn ; Infant ; Hearing Loss/genetics* ; Retrospective Studies ; Child, Preschool ; Child ; Genotype ; Connexins/genetics* ; Mutation

Objective:To evaluate the clinical efficacy and prognosis of mechanical lithotripsy, laser lithotripsy under direct peroral cholangioscopy, and their combination in the treatment of difficult common bile duct (CBD) stones.Methods:Clinical data of 345 patients with difficult CBD stones treated at the Department of Hepatobiliary Surgery, Shandong Provincial Third Hospital, Shandong University, between January 2020 and December 2024 were retrospectively analyzed, including 176 males and 169 females, aged (71.2±14.2) years. Patients were categorized into three groups based on the lithotripsy technique used: mechanical lithotripsy group ( n=275), laser lithotripsy group under direct peroral cholangioscopy ( n=34), and combined lithotripsy group ( n=35). Operative time, hospitalization costs, stone clearance rate, and postoperative complications were recorded. Follow-ups were conducted through outpatient visits and telephone reviews to monitor stone recurrence. Propensity score matching (PSM) at a 1: 3 nearest-neighbor ratio with a caliper of 0.02 was performed, using lithotripsy method as the dependent variable, and age, sex, stone size, and bile duct diameter as independent variables, resulting in well-balanced mechanical and laser lithotripsy groups. Kaplan-Meier survival analysis was used to assess recurrence-free survival, with comparisons performed using the log-rank test. Results:Before PSM, there were significant differences in age, sex, stone length, and bile duct diameter between the groups (all P<0.05). After PSM, 40 patients were included in the mechanical lithotripsy group, 34 in the laser group, and 35 in the combined group, with no significant differences in baseline or preoperative clinical characteristics (all P>0.05). The combined group had a significantly longer operative time compared to the mechanical group [71.0 (66.0, 92.0) min vs. 50.5 (40.4, 56.5) min, Z=-5.02, P<0.001] and the laser group [71.0 (66.0, 92.0) min vs. 53.0 (26.5, 73.5) min, Z=-2.61, P=0.001]. The laser group also had a longer operative time than the mechanical group [53.0 (26.5, 73.5) min vs. 50.5 (40.4, 56.5) min, Z=-2.27, P=0.023]. Hospitalization costs were significantly higher in the combined group compared to the mechanical group [43 000(33 000, 50 000) yuan vs. 30 000(26 000, 37 000) yuan; Z=-3.43, P<0.001]. The single-session stone clearance rates were 80.0% (32/40) for the mechanical group, 85.3% (29/34) for the laser group, and 62.9% (22/35) for the combined group. Postoperative complication rates were 20.0% (8/40), 11.7% (4/34), and 11.4% (4/35), respectively, with no statistically significant differences among the three groups (all P>0.05). There were also no significant differences in cumulative recurrence-free survival among the groups ( χ2=0.06, P=0.970). Conclusions:For endoscopic management of difficult CBD stones, combined lithotripsy is associated with longer operative time and higher hospitalization costs compared to mechanical and laser lithotripsy alone. Laser lithotripsy also requires more operative time than mechanical lithotripsy. However, the three lithotripsy strategies show no significant differences in postoperative complications or cumulative recurrence-free survival.