OBJECTIVE: To investigate the current status of clinical genetics specialization development and the diagnostic and therapeutic capabilities for hereditary diseases across medical institutions in Shanghai, and to assess the necessity and feasibility of establishing training bases for clinical genetics specialists. METHODS: By employing a cross-sectional survey design, the Clinical Genetics Committee of Shanghai Medical Association has conducted questionnaire surveys from March to April 2025 across 54 healthcare institutions in Shanghai (including 33 tertiary hospitals and 21 secondary hospitals). The survey involved administrative departments and medical personnel from 15 clinical specialties. The survey has covered current genetic disease diagnosis and treatment practices, relevant and specialised disease types, genetic department establishment, testing capabilities, personnel teams, and training requirements. RESULTS: The results revealed that 78.0% of clinical departments surveyed had treated patients with hereditary disorders. Shanghai possesses diagnostic and therapeutic expertise for over 95% of hereditary diseases listed in its rare disease catalogue, reflecting both the practical clinical demand for such conditions and the city's overall diagnostic and therapeutic strengths in this field. Nevertheless, significant disparities exist in the development of genetics departments across different tiers of healthcare institutions. Resources for genetic testing capabilities (including molecular, cellular, and biochemical testing) are also unevenly distributed across different tiers of hospitals. The survey further revealed that only 26.0% of departments believe that their current physician structure fully meets the diagnostic and treatment demands. Over 90% of departments consider standard training for clinical genetic specialists necessary, with 74.0% expressing willingness to participate in establishing training bases. Based on above findings and thorough deliberation, the Clinical Genetics Committee of the Shanghai Medical Association proposes advancing specialist training and discipline development through establishing a standard training system. The committee has drafted a three-year training protocol featuring a "joint training"-centered model, recommending a pilot-first, dynamically optimized strategy for steadily advancing training base development. CONCLUSION Shanghai faces substantial demand for genetic disease diagnosis and treatment, yet exhibits shortcomings in clinical genetics specialization development, resource allocation, and talent pipeline cultivation. To establish a standard training system holds significant practical importance and is underpinned by a broad demand.
Humans ; China ; Surveys and Questionnaires ; Genetic Diseases, Inborn/genetics* ; Cross-Sectional Studies ; Genetics, Medical/education* ; Genetic Testing

Objective: To optimize the technology and process for recovering immune cells from discarded leukocyte filter through reverse flushing, and to investigate the effects of different reverse flushing solutions and physical conditions (temperature, soaking, and ultrasound) on the cell recovery rate. Additionally, to evaluate the feasibility and quality of isolating and culturing NK cells from the recovered cells. Methods: Normal saline, PBS, and PBS-EDTA were used as reverse flushing solutions to backwash the filter, and the white blood cell recovery rates were compared. The reverse flushing efficiency was optimized by adjusting the temperature of the reverse flushing solution, filter soaking treatment and ultrasonic treatment. The Ficoll separation method was employed to isolate peripheral blood mononuclear cells (PBMCs) from the elution solution and conduct in vitro expansion and culture. Lymphocyte subsets were identified and quantified using flow cytometry. Results: The physiological saline served as the backflush fluid was the most stable. The reverse temperature had a significant impact on the recovery rate of white blood cell subgroups. The recovery rates of lymphocytes and monocytes at 37℃ were significantly higher than those at 4℃ [(35.56±5.55)% vs (22.88±0.31)%, (32.42±4.82)% vs (8.00±1.6)%, t=5.581 and 5.758, respectively, both P<0.001)], and at 25℃ [(35.56±5.55)% vs (22.42±6.13)%, t=5.904, P<0.001; (32.4±4.82)% vs (21.11±1.23)%, t=2.629, P=0.030)]. The recovery rates of white blood cell subgroups after 37℃ immersion were higher than those after direct reverse flow at 37℃, but the difference was not statistically significant (P>0.05). The recovery rate of cells after ultrasonic immersion at 25℃ was further increased compared to that after immersion at 25℃, but the difference was not statistically significant (P>0.05). The purity of the NK cells obtained through sorting reached (94.2±4.5)%, and the viability was (94.8±0.9)%. After 15 days of culture, the cell count was expanded approximately 450 times, yielding more than 10 /mL highly active NK cells. Conclusion: By optimizing the backflushing process, immune cells can be efficiently recovered from discarded filters. The 37℃ backflow liquid has the best backflow effect; soaking and ultrasonic treatment can be used as auxiliary methods to improve recovery rates. The NK cells from this source have high purity, good activity, and strong expansion ability, and can be used as a reliable cell source for immunotherapy research, and in line with the circular economy concept of resource reuse.

AIM:To investigate the dynamic changes in ocular surface function after eyelid malignant tumor resection combined with Hughes flap reconstruction.

METHODS:A prospective self-controlled study was conducted. Patients who underwent eyelid malignant tumor resection combined with Hughes tarsoconjunctival flap reconstruction(all tumors occured in the lower eyelid)at Shanxi Eye Hospital from January 2021 to December 2023 were enrolled. The contralateral healthy eye served as control. Ocular surface examinations were performed preoperatively and at 3, 6, 9, and 12 mo postoperatively, including ocular surface disease index(OSDI), tear meniscus height(TMH), non-invasive average tear film break-up time(NiaBUT), tear film lipid layer grading, upper and lower eyelid meibomian gland dropout grading, corneal fluorescein staining, and eye redness index.

RESULTS:A total of 36 patients were enrolled. Two patients were lost to follow-up because they did not complete the scheduled follow-up period, leaving 34 patients with complete data for analysis. The cohort comprised 10 men(29%)and 24 women(71%), with a mean age of 70.08±10.92 y(range 33-81 y). Histopathological diagnoses included basal cell carcinoma(BCC)in 22 eyes(65%), sebaceous gland carcinoma(SGC)in 10 eyes(29%), and squamous cell carcinoma(SCC)in 2 eyes(6%). Tumor size ranged from 4 to 30 mm(mean 12.64±0.31 mm). According to the AJCC staging system, 14 eyes(41%), 18 eyes(53%), and 2 eyes(6%)were classified as T1b, T2b, and T3b, respectively. Preoperatively, OSDI scores were higher in surgical eyes than in contralateral healthy eyes(28.74±13.72 vs 17.47±3.02, P<0.05). OSDI scores in surgical eyes further increased at 6, 9, and 12 mo postoperatively, peaking at 9 mo(all P<0.05), whereas no significant postoperative changes were observed in contralateral healthy eyes. Compared with contralateral healthy eyes, TMH in surgical eyes increased significantly at 3 mo postoperatively and remained elevated thereafter despite a slight decline(all P<0.05). NiaBUT in surgical eyes was shorter than the preoperative value from 3 mo postoperatively onward and was lower than that in contralateral healthy eyes(all P<0.05). Eye redness index in surgical eyes were consistently higher than those in contralateral healthy eyes(all P<0.001). Tear film lipid layer grade and meibomian gland dropout grades in both the upper and lower eyelids were significantly worsened postoperatively compared with baseline(all P<0.001). Corneal fluorescein staining grades were higher in surgical eyes than in contralateral healthy eyes preoperatively(P<0.001)and were higher at 3, 6, 9, and 12 mo postoperatively than at baseline(all P<0.001).

CONCLUSION: After lower eyelid malignant tumor resection combined with Hughes flap reconstruction, evaporative dry eye is the predominant phenotype. Meibomian gland dropout in both upper and lower eyelids appears irreversible within 12 mo postoperatively, and corneal epithelial damage and ocular surface inflammation persist over time.

Background: Maturity-onset diabetes of the young (MODY) due to variants of hepatocyte nuclear factor 1-beta (HNF1β) (MODY5) has not been well studied in the Chinese population. This study aimed to estimate its prevalence and evaluate the application of a clinical screening method (Faguer score) in Chinese early-onset diabetes (EOD) patients. Methods: Among 679 EOD patients clinically diagnosed with type 2 diabetes mellitus (age at diagnosis ≤40 years), the exons of HNF1β were sequenced. Functional impact of rare variants was evaluated using a dual-luciferase reporter system. Faguer scores ≥8 prompted multiplex ligation-dependent probe amplification (MLPA) for large deletions. Pathogenicity of HNF1β variants was assessed following the American College of Medical Genetics and Genomics (ACMG) guidelines. Results: Two rare HNF1β missense mutations (E105K and G454R) were identified by sequencing in five patients, showing functional impact in vitro. Another patient was found to have a whole-gene deletion by MLPA in 22 patients with the Faguer score above 8. Following ACMG guidelines, six patients carrying pathogenic or likely pathogenic variant were diagnosed with MODY5. The estimated prevalence of MODY5 in Chinese EOD patients was approximately 0.9% or higher. Conclusion MODY5 is not uncommon in China. The Faguer score is helpful in deciding whether to perform MLPA analysis on patients with negative sequencing results.

As new evidence emerges, treatment strategies toward the functional cure of chronic hepatitis B are evolving. In 2019, a panel of national hepatologists published a Consensus Statement on the functional cure of chronic hepatitis B. Currently, an international group of hepatologists has been assembled to evaluate research since the publication of the original consensus, and to collaboratively develop the updated statements. The 2.0 Consensus was aimed to update the original consensus with the latest available studies, and provide a comprehensive overview of the current relevant scientific literatures regarding functional cure of hepatitis B, with a particular focus on issues that are not yet fully clarified. These cover the definition of functional cure of hepatitis B, its mechanisms and barriers, the effective strategies and treatment roadmap to achieve this endpoint, in particular new surrogate biomarkers used to measure efficacy or to predict response, and the appropriate approach to pursuing a functional cure in special populations, the development of emerging antivirals and immunomodulators with potential for curing hepatitis B. The statements are primarily intended to offer international guidance for clinicians in their practice to enhance the functional cure rate of chronic hepatitis B.

Objective:To investigate factors influencing frequent episodes (≥ 4 episodes within 1 year) in children with moderate-to-severe atopic dermatitis (AD) in China.Methods:A national multicenter cross-sectional study was conducted. Patients under the age of 18 years diagnosed with moderate-to-severe AD were enrolled at dermatology clinics in 18 medical institutions across 12 provinces and municipalities in China between June 12 and August 8, 2023. At the time of the visit, their guardians completed a structured questionnaire covering demographic characteristics, clinical features of AD, personal and family history, factors associated with frequent episodes of moderate-to-severe AD, compliance with treatment, and disease awareness. Statistical analyses included t tests, one-way analysis of variance, rank-sum tests, and chi-square tests, with multiple-response analysis applied for multiple-choice questions. Results:A total of 965 valid questionnaires were collected, and 965 children with moderate-to-severe AD were included. Among them, there were 531 males and 434 females, 678 (70.3%) were aged 2 - < 12 years, 837 (86.7%) were from urban areas, the age at onset was 2.47 ± 3.03 years, and the median frequency of AD episodes in the past year was 4 times. These children were divided into 2 groups based on the median episode frequency: < 4-episode group (439 cases, 45.5%) and ≥ 4-episode group (526 cases, 54.5%). Compared with the < 4-episode group, children in the ≥ 4-episode group showed younger ages at onset (2.22 ± 2.98 years vs. 2.76 ± 3.06 years, P = 0.006) and higher proportions of patients with comorbid allergic diseases in both the children themselves (82.9% [436/526] vs. 69.7% [306/439], χ2 = 23.42, P < 0.001) and their relatives (66.0% [347/526] vs. 57.4% [252/439], χ2 = 7.46, P = 0.006). Children in the ≥ 4- episode group also had higher monthly usage of moisturizers (150 [30, 300] g vs. 60 [6, 200] g) and daily frequency of moisturizer use, greater disease awareness, but more severe fear of medication use (all P < 0.05). The region and the human development index level were both significantly associated with the episode frequency (both P < 0.001), with the highest proportion of children from South China in the ≥ 4- episode group (36.3%, 191/526). Children in the ≥ 4-episode group also had a longer duration of topical glucocorticoid use than those in the < 4-episode group ( Z = -2.21, P = 0.027). External triggers associated with AD episodes mainly included heat exposure (50.36%, 486/965), hot water bathing (40.73%, 393/965), seafood (23.52%, 227/965), and dust mites (33.37%, 322/965) . Conclusion:In children with moderate-to-severe AD in China, factors influencing frequent episodes may include residence in southern or economically developed regions, earlier age at onset, having a personal or family history of allergic diseases, and fear of medication use.

Objective:Patients with metastatic urothelial carcinoma(mUC)usually receive platinum-based chemotherapy as the first-line treatment.Recently,antibody-drug conjugates(ADCs)combined with programmed death-1 antibody(PD-1 antibody)have shown promising efficacy and safety in the treatment of mUC.HER2 positivity is associated with poor prognosis,patients can benefit from anti-HER2 ADC therapy such as disitamab vedotin(RC48-ADC).This study aimed to evaluate the efficacy and safety of RC48-ADC combined with tislelizu-mab in treatment-na?ve patients with mUC.Methods:A retrospective analysis was performed on 70 mUC patients treated between July 2022 and December 2023,including 30 patients receiving RC48-ADC combined with tislelizumab(DT group)and 40 patients receiving gem-citabine plus cisplatin(GC group).Primary endpoints included objective response rate(ORR),disease control rate(DCR),progression-free survival(PFS),overall survival(OS),and treatment-related adverse events(TRAEs).Results:The ORR(73.3%vs.47.5%)and DCR(86.7%vs.62.5%)were significantly higher in the DT group compared to the GC group(P<0.05).The DT group also demonstrated longer PFS(10.98 months vs.7.67 months,P<0.005)and prolonged OS(median OS:not reached vs.11.34 months).The most common TRAEs included myel-osuppression,gastrointestinal and hepatobiliary toxicities,fatigue,alopecia,and rash.No grade≥3 TRAEs or treatment-related deaths were reported.Conclusions:RC48-ADC combined with tislelizumab demonstrated superior efficacy and favorable safety in treatment-na?ve pa-tients with mUC,supporting its potential as a first-line therapeutic option.

AIM To investigate the impact of varying dosages of Supplemented Wendan Decoction on the PI3K/Akt/FOXO1 glycolipid metabolic pathway in 3T3-L1 adipocytes.METHODS The CCK-8 assay was used to determine the concentration of Supplemented Wendan Decoction-medicated serum.The mature adipocytes differentiated from 3T3-L1 preadipocytes after induction were further divided into the blank control group,the model group,the rosiglitazone group(10 mg/L),and the Supplemented Wendan Decoction groups(5％,10％,and 20％),followed by the sample collections after 48 hours of treatment.Oil red O staining quantified lipid accumulation in 3T3-L1 adipocytes;extracellular glucose levels were measured using glucose oxidase(GOD)assay;RT-qPCR analyzed mRNA expressions of IRS-1,PI3K,Akt,GLUT4,IL-6,TNF-α and IL-1β;Western blot assessed protein expressions of INSR,IRS-1,PI3K-p85,Akt,FOXO1 and GLUT4.RESULTS No significant changes in cell viability(P＞0.05)were observed in 3T3-L1 preadipocytes exposed to serum containing supplemented Wendan Decoction at different concentrations for 24,48,or 72 hours.The 3T3-L1 preadipocytes held the capacity to differentiate into mature adipocytes within a 14-day induction period.Compared to the model group,all supplemented Wendan Decoction groups exhibited reduced lipid accumulation in adipocytes and downregulated mRNA expression of IRS-1,IL-6,TNF-α and IL-1β(P＜0.01);the low-dose group demonstrated increased mRNA expressions of PI3K and GLUT4(P＜0.05,P＜0.01),alongside elevated protein expressions of INSR,IRS-1,PI3K-p85,Akt and GLUT4(P＜0.05,P＜0.01);the medium-dose group showed enhanced GLUT4 mRNA expression,and upregulated protein expressions of INSR and FOXO1(P＜0.01).After 24 hours intervention,the high-dose Supplemented Wendan Decoction group exhibited increased glucose consumption in adipocytes(P＜0.01),and elevated protein expression of INSR,Akt and FOXO1(P＜0.05,P＜0.01).CONCLUSION Supplemented Wendan Decoction reduces lipid accumulation in adipocytes,regulates glucose and lipid metabolism,and promotes metabolic homeostasis through PI3K/Akt/FOXO1 signaling pathway.

Objective To investigate the role of additional platelet protein kinase C epsilon(PKCε)assay in risk stratification and additional diagnostic testing in elderly patients with acute chest pain and normal hypersensitive cardiac troponin I(hs-cTnI)level.Methods A prospective trial was conducted on 220 consecutive elderly patients with angina-like acute chest pain and normal hs-cTnI level in our hospital from April 2022 to April 2023.According to whether major adverse cardiovascular events(MACE)occurred during the 1-year follow-up period,they were assigned in-to MACE group(24 cases)and non-MACE group(196 cases).Their general clinical data and platelet PKCε expression were compared between the two groups.Kaplan-Meier survival curve was plotted to analyzed the event-free time between the platelet PKCε expression＜2.05％group and the≥2.05％group.Multivariate Cox regression analysis was used to investigate the impact of baseline data on the occurrence of MACE during the 1-year follow-up period.Decision tree model was employed to analyze the influencing factors for MACE.Results During 1-year follow-up,24 patients(10.91％)developed MACE.The platelet PKCε expression was significantly higher in the MACE group than the non-MACE group[9.25％(6.60％,15.25％)vs 1.70％(0.80％,3.25％),Z=-7.596,P＜0.01].Kaplan-Meier survival curve analysis showed that the patients in the PKCε＜2.05％group had no events within 360 d,while those in the ≥2.05％group had an average event-free time of 297.13±12.77 d,with a statistical difference(log rank Chi-square=27.051,P=0.000).Multivariate Cox regression analysis showed that highly suspicious angina,hs-cTnI and platelet PKCε were independent risk factors for MACE within 1 year of follow-up(HR=3.011,95％CI:1.049-8.641,P=0.040;HR=1.216,95％CI:1.044-1.418,P=0.012;HR=1.160,95％CI:1.104-1.218,P=0.000).In the decision tree model,the 1-year incidence of MACE was notably higher in the platelet PKCε≥2.05％group than the＜2.05％group(P=0.011).The probability of severe coronary artery stenosis or myocardial ischemia in the platelet PKCε≥2.05％group was remarkably higher than the＜2.05％group(P=0.006).Conclusion In elderly patients suffering from acute chest pain with normal hs-cTnI level,additional platelet PKCε detection appears to have a good prognostic gain.

Due to the poor ionization efficiency and the weak mass spectrometry(MS)intensity of weakly polar substances,direct analysis using the traditional electrospray ionization mass spectrometry(ESI-MS)is a big challenge.In this study,a novel rapid on-site detection method of four prohibited sex hormones in cosmetics was proposed using online derivatization strategy coupled with a miniature mass spectrometer.The target substances in the samples were extracted by a custom-made polyaniline/multi-walled carbon nanotube solid-phase microextraction(SPME)probe.The stirring speed was 200 r/min,the extraction temperature was 40℃,and the extraction time was 2 min.A pulled dual-channel θ borosilicate glass capillary emitter was used as the nano-ESI ion source.The SPME probe was inserted into the channel containing methanol in theθborosilicate glass capillary.When the spray voltage was applied,the four sex hormones were desorbed and formed spray microdroplets,which then collided with the hydroxylamine microdroplets generated from the other channel.The microdroplets of reaction product entered into the miniature mass spectrometer for direct analysis.The limits of detection(LOD)and limits of quantification(LOQ)for the four sex hormones were 10-20 ng/mL and 20-50 ng/mL,respectively.The recoveries were from 84.6%to 107.8%with the relative standard deviations(RSD)from 4.1%to 11.6%.Compared to detection without derivatization,the MS signals of the four target substances were increased by 3 to 15 times.This method was simple,rapid,highly efficient and sensitive,and suitable for on-site rapid analysis of weakly polar sex hormones in cosmetics.