Objective: To propose an improved method for constructing the internal quality control (IQC) framework for ELISA assays and validate its efficacy by statistically analyzing IQC data from nine blood center laboratories. Methods: 1) IQC data was collected from nine blood centers and analyzed using a domestic HBsAg ELISA detection kit as an example. 2) Differences between IQC values across batches within Blood Center 1 were assessed. 3) Statistical analyses were performed on batch usage, number of batches used, days of use, number of QC points, batch-specific means, and coefficients of variation (CV) across all nine centers. 4) Using the improved construction method for IQC framework, provisional and permanent frames were established for batches within Blood Center 1 and Blood Center 9, followed by outlier determination. Results: 1) Statistically significant differences were observed in IQC data between batches within Blood Center 1 (P<0.01). It is recommended that both the control material/reagents and the control chart framework be replaced simultaneously. 2) There were substantial differences among 9 blood centers regarding the control material/reagent lot numbers used, the number of QC runs per batch, and the QC values for identical lots. Therefore, individual laboratories should establish their own IQC chart frameworks. 3) The improved IQC framework construction method for ELISA assays is as follows: provisional frames are established via frame-shifting, using the pre-experimental mean and cumulative coefficient of variation (CV) from the preceding batch. For batches used >20 days with >20 QC points, permanent frames are constructed by aggregating in-control data accumulated over ≥20 days with ≥20 points to calculate cumulative mean and standard deviation. The provisional and permanent frames constructed by this method identified all 26 extreme outliers across Blood Centers 1 and 9 as out-of-control. Among the 218 general outliers, 10 were classified as normal by the provisional frames, while the remainder were designated as warnings or out-of-control. This method effectively monitors assay stability. Conclusion: Based on the statistical analysis of IQC practices across blood centers of varying scales, combined with the inherent characteristics of ELISA assays and the batch-to-batch instability of reagents/QC materials, it is recommended to reconstruct QC charts upon lot changes. The proposed method—utilizing frame-shifting for provisional frames and establishing permanent frames based on cumulative data—is applicable to blood center laboratories of differing sizes and effectively monitors the stability of the ELISA assay process.

Background and purpose:Internal mammary sentinel lymph node biopsy(IMSLNB)is a minimally invasive diagnostic technique for regional lymph nodes in breast cancer,which can provide accurate lymph staging and guide adjuvant treatment decision,but its clinical application has been controversial.The purpose of this study was to investigate the prognosis of IMSLNB in early breast cancer.Methods:In this study,a retrospective cohort of 7 949 patients with breast cancer from January 1,2016 to December 31,2021 was analyzed.After applying propensity score matching,the patients were divided into IMSLNB group and no-IMSLNB group,and the regional recurrence-free survival(RRFS),local recurrence-free survival(LRFS),distant metastasis-free survival(DMFS),disease-free survival(DFS)and overall survival(OS)of the two groups were compared.This study was approved by the Clinical Research Ethics Committee of the Affiliated Tumor Hospital of Shandong First Medical University(approval number:SDTHEC20130324).Results:A total of 990 patients were included in the final analysis(330 in the IMSLNB group and 660 in the no-IMSLNB group).IMSLN metastasis was found in 54 patients in the IMSLNB group,including 47 patients with axillary lymph node(ALN)metastasis and 7 patients with IMSLN metastasis only.The IMSLN transfer rate was 16.4%.The median follow-up of 41 months showed that the IMSLNB group demonstrated better 3-year DFS[98.4%vs 94.2%,hazard ratio(HR)=0.509;95%CI:0.312-0.828,P=0.007]and 5-year DFS(92.5%vs 87.3%,HR=0.214,95%CI:0.206-0.222,P=0.011)compared with no-IMSLNB group.However,no significant differences were observed in 3-year OS(99.1%vs 99.4%,HR=0.618,95%CI:0.231-1.655,P=0.338)or 5-year OS(98.5%vs 99.1%,HR=0.52,95%CI:0.51-0.53,P=0.392)between the two groups.The 3-year RRFS in the IMSLNB group was better compared with the no-IMSLNB group(99.09%vs 97.73%,HR=0.066;95%CI:0.061-0.071,P=0.048),while no significant differences were observed in 3-year LRFS(99.70%vs 98.19%,HR=0.209;95%CI:0.201-0.217,P=0.130)or DMFS(95.76%vs 96.06%,HR=0.865,95%CI:0.858-0.872,P=0.820)between the two groups.The exploratory subgroup analysis of DFS revealed that patients in the following subgroups could significantly benefit from IM-SLNB(P＜0.05):diagnosis age(≤50 years),premenopausal status,BMI(≤24),lymphovascular invasion(LVI,present),tumor location(lateral),molecular subtype[hormone receptor positive(HR+)/human epidermal growth factor receptor 2 negative(HER2-)],histological type(invasive ductal carcinoma),and axillary lymph node status(positive).Conclusion:IMSLNB can provide more accurate regional lymph node staging for early breast cancer,help optimize adjuvant radiotherapy strategies,and improve patients'DFS and RRFS.It can be promoted as a minimally invasive staging technique for regional lymph nodes.

Background and purpose:Internal mammary sentinel lymph node biopsy(IMSLNB)is a minimally invasive diagnostic technique for regional lymph nodes in breast cancer,which can provide accurate lymph staging and guide adjuvant treatment decision,but its clinical application has been controversial.The purpose of this study was to investigate the prognosis of IMSLNB in early breast cancer.Methods:In this study,a retrospective cohort of 7 949 patients with breast cancer from January 1,2016 to December 31,2021 was analyzed.After applying propensity score matching,the patients were divided into IMSLNB group and no-IMSLNB group,and the regional recurrence-free survival(RRFS),local recurrence-free survival(LRFS),distant metastasis-free survival(DMFS),disease-free survival(DFS)and overall survival(OS)of the two groups were compared.This study was approved by the Clinical Research Ethics Committee of the Affiliated Tumor Hospital of Shandong First Medical University(approval number:SDTHEC20130324).Results:A total of 990 patients were included in the final analysis(330 in the IMSLNB group and 660 in the no-IMSLNB group).IMSLN metastasis was found in 54 patients in the IMSLNB group,including 47 patients with axillary lymph node(ALN)metastasis and 7 patients with IMSLN metastasis only.The IMSLN transfer rate was 16.4%.The median follow-up of 41 months showed that the IMSLNB group demonstrated better 3-year DFS[98.4%vs 94.2%,hazard ratio(HR)=0.509;95%CI:0.312-0.828,P=0.007]and 5-year DFS(92.5%vs 87.3%,HR=0.214,95%CI:0.206-0.222,P=0.011)compared with no-IMSLNB group.However,no significant differences were observed in 3-year OS(99.1%vs 99.4%,HR=0.618,95%CI:0.231-1.655,P=0.338)or 5-year OS(98.5%vs 99.1%,HR=0.52,95%CI:0.51-0.53,P=0.392)between the two groups.The 3-year RRFS in the IMSLNB group was better compared with the no-IMSLNB group(99.09%vs 97.73%,HR=0.066;95%CI:0.061-0.071,P=0.048),while no significant differences were observed in 3-year LRFS(99.70%vs 98.19%,HR=0.209;95%CI:0.201-0.217,P=0.130)or DMFS(95.76%vs 96.06%,HR=0.865,95%CI:0.858-0.872,P=0.820)between the two groups.The exploratory subgroup analysis of DFS revealed that patients in the following subgroups could significantly benefit from IM-SLNB(P＜0.05):diagnosis age(≤50 years),premenopausal status,BMI(≤24),lymphovascular invasion(LVI,present),tumor location(lateral),molecular subtype[hormone receptor positive(HR+)/human epidermal growth factor receptor 2 negative(HER2-)],histological type(invasive ductal carcinoma),and axillary lymph node status(positive).Conclusion:IMSLNB can provide more accurate regional lymph node staging for early breast cancer,help optimize adjuvant radiotherapy strategies,and improve patients'DFS and RRFS.It can be promoted as a minimally invasive staging technique for regional lymph nodes.

Objective:To summarize the clinical and genetic characteristics of late-onset facioscapulohumeral muscular dystrophy type 1 (FSHD1) patients, and to compare the differences between late-onset and classic-onset FSHD1 patients.Methods:A retrospective analysis was conducted on the clinical and genetic data of genetically confirmed late-onset FSHD1 patients (age at onset30 years) between January 2007 and June 2024 from the Department of Neurology of Peking University First Hospital and the First Affiliated Hospital of Fujian Medical University. Classic-onset FSHD1 patients (10 yearsage at onset≤30 years) were matched 1∶1 according to sex and disease duration for comparison. The demographic information, the number of D4Z4 repeat units, the distal D4Z4 methylation levels, FSHD Clinical Score (CS), Clinical Severity Score (CSS), and Age-Corrected Clinical Severity Score (ACSS) of these patients were collected. Survival analysis was performed to compare the outcome of lower extremity involvement between late-onset and classic-onset FSHD1 patients. The correlation of the number of D4Z4 repeat units and D4Z4 methylation level with CS and ACSS was analyzed in late-onset FSHD1 patients.Results:A total of 61 patients with late-onset FSHD1 were enrolled, 33 (54.1%) of whom are female, with an age of 54.0 (46.0, 62.0) years and a disease duration of 14.0 (5.5, 22.5) years. Compared to classic-onset FSHD1 patients, late-onset patients exhibited significantly lower CS [7.0 (5.6, 8.4) vs 6.0 (4.4, 7.7), U=1 416.000, P=0.013], CSS [3.0 (2.8, 3.3) vs 3.0 (2.0, 4.0), U=2 352.000, P=0.010], and ACSS [189.2 (137.1, 241.3) vs 96.8 (61.3, 132.2), U=3 225.500, P0.001], and higher proportion of patients with limb girdle involvement but no facial muscle involvement [18.0% (11/61) vs 6.6% (4/61), χ2=3.725, P=0.054]. Kaplan-Meier survival analysis showed that the onset age of lower extremity involvement in late-onset patients (45 years, 95% CI 42-48 years) was significantly higher than that in classic-onset patients (24 years, 95% CI 21-27 years, χ2=61.012, P0.001). The duration from symptom onset to lower extremity involvement in late-onset patients (15 years, 95% CI 10-20 years) was significantly longer than that in classic-onset patients (8 years, 95% CI 3-13 years, χ2=9.105, P=0.003). Late-onset FSHD1 patients carried higher average distal D4Z4 methylation levels compared to those with classic-onset FSHD1 [46.68% (40.79%,52.57%) vs 41.02% (34.03%,48.00%), U=1 378.500, P=0.014]. Among late-onset FSHD1 patients, cytosine-phosphate-guanine 6 (CpG6) methylation levels were significantly negatively correlated with ACSS ( r=-0.278, P=0.025); the number of D4Z4 repeat units were significantly negatively correlated with ACSS ( r=-0.272, P=0.034);CpG6 methylation levels were significantly negatively correlated with CS ( r=-0.441, P=0.003), while no correlation was found between number of D4Z4 repeat units and CS ( r=-0.161, P=0.310). Conclusions:Compared with classic-onset FSHD1 patients, late-onset FSHD1 patients are associated with a higher degree of distal D4Z4 methylation, along with a milder muscle weakness phenotype, slower disease progression and a higher proportion of cases without facial muscle involvement. The age at onset can be used as a marker of the severity and prognosis in FSHD1.

Background and purpose:With the progressive development of breast cancer surgery toward more individualized and minimally invasive approaches,sentinel lymph node biopsy(SLNB)has replaced axillary lymph node dissection(ALND)as the standard method for axillary management in certain early-stage breast cancer patients.However,there is ongoing debate in clinical practice regarding whether regional lymph node irradiation(RNI)is necessary for patients with sentinel lymph node(SLN)positive status who have not undergone ALND.This study aimed to analyze the clinicopathological features and survival prognosis of patients with SLN-positive status who did not undergo ALND,evaluate the clinical application value of RNI,and provide evidence to support clinical treatment decisions for this group of patients.Methods:This single-center retrospective study screened breast cancer patients who underwent SLNB at Shandong Cancer Hospital from September 1,2014,to August 31,2023.All patients signed informed consent for treatment.Based on whether postoperative radiotherapy included regional lymph node irradiation(internal mammary and/or axillary and/or supra-/infra-clavicular fields),patients were divided into the RNI group and the no-RNI group for follow-up.Additionally,patients were further divided into multiple subgroups based on factors such as the type of breast surgery,tumor molecular subtype,and histological grade,to compare the clinical value of RNI among subgroups.The primary endpoint was locoregional recurrence-free survival(LRRFS),and the secondary endpoints included invasive disease-free survival(iDFS)and overall survival(OS).The Strengthening the Reporting of Observational Studies in Epidemiology(STROBE)checklist was followed for this study.Results:Clinical data of 8 328 breast cancer patients'were screened for this study,and after applying inclusion and exclusion criteria,356 patients were included in the analysis,with 186 in the RNI group and 170 in the no-RNI group.There were no significant differences between the two groups in terms of age,body mass index(BMI),menopausal status,tumor location,pathological type,histological grade,vascular invasion,estrogen receptor(ER)and progesterone receptor(PR)status,and human epidermal growth factor receptor 2(HER-2)expression(P＞0.05).However,the number of positive SLNs,T stage,and the proportion of patients undergoing total mastectomy(TM)were significantly higher in the RNI group than in the no-RNI group(P=0.006,P=0.043,P＜0.001).After a median follow-up of 38 months,no recurrence or metastasis was observed in the RNI group,while the recurrence and metastasis rate in the no-RNI group was 3.5%(6/170).Of these,4 cases had local regional recurrence,and 2 had distant metastasis.The RNI group showed superior iDFS compared to the no-RNI group(P=0.017),however there was no statistically significant difference in LRRFS and OS(P=0.051 and P=0.356).Exploratory subgroup analysis indicated that patients with tumor diameter＞2 cm(P=0.033)and triple-negative molecular(TNBC)(P=0.020)might benefit from RNI treatment in terms of LRRFS.Conclusion:For certain high-risk patients,such as those with larger tumor diameter,TNBC,or high non-SLN metastatic risk,RNI still plays an important role in reducing the risk of recurrence and metastasis in breast cancer.In clinical practice,an individualized RNI strategy should be developed based on the patient's residual lymph node tumor load,biological behavior of the tumor,and surgical method.

Background and purpose:With the progressive development of breast cancer surgery toward more individualized and minimally invasive approaches,sentinel lymph node biopsy(SLNB)has replaced axillary lymph node dissection(ALND)as the standard method for axillary management in certain early-stage breast cancer patients.However,there is ongoing debate in clinical practice regarding whether regional lymph node irradiation(RNI)is necessary for patients with sentinel lymph node(SLN)positive status who have not undergone ALND.This study aimed to analyze the clinicopathological features and survival prognosis of patients with SLN-positive status who did not undergo ALND,evaluate the clinical application value of RNI,and provide evidence to support clinical treatment decisions for this group of patients.Methods:This single-center retrospective study screened breast cancer patients who underwent SLNB at Shandong Cancer Hospital from September 1,2014,to August 31,2023.All patients signed informed consent for treatment.Based on whether postoperative radiotherapy included regional lymph node irradiation(internal mammary and/or axillary and/or supra-/infra-clavicular fields),patients were divided into the RNI group and the no-RNI group for follow-up.Additionally,patients were further divided into multiple subgroups based on factors such as the type of breast surgery,tumor molecular subtype,and histological grade,to compare the clinical value of RNI among subgroups.The primary endpoint was locoregional recurrence-free survival(LRRFS),and the secondary endpoints included invasive disease-free survival(iDFS)and overall survival(OS).The Strengthening the Reporting of Observational Studies in Epidemiology(STROBE)checklist was followed for this study.Results:Clinical data of 8 328 breast cancer patients'were screened for this study,and after applying inclusion and exclusion criteria,356 patients were included in the analysis,with 186 in the RNI group and 170 in the no-RNI group.There were no significant differences between the two groups in terms of age,body mass index(BMI),menopausal status,tumor location,pathological type,histological grade,vascular invasion,estrogen receptor(ER)and progesterone receptor(PR)status,and human epidermal growth factor receptor 2(HER-2)expression(P＞0.05).However,the number of positive SLNs,T stage,and the proportion of patients undergoing total mastectomy(TM)were significantly higher in the RNI group than in the no-RNI group(P=0.006,P=0.043,P＜0.001).After a median follow-up of 38 months,no recurrence or metastasis was observed in the RNI group,while the recurrence and metastasis rate in the no-RNI group was 3.5%(6/170).Of these,4 cases had local regional recurrence,and 2 had distant metastasis.The RNI group showed superior iDFS compared to the no-RNI group(P=0.017),however there was no statistically significant difference in LRRFS and OS(P=0.051 and P=0.356).Exploratory subgroup analysis indicated that patients with tumor diameter＞2 cm(P=0.033)and triple-negative molecular(TNBC)(P=0.020)might benefit from RNI treatment in terms of LRRFS.Conclusion:For certain high-risk patients,such as those with larger tumor diameter,TNBC,or high non-SLN metastatic risk,RNI still plays an important role in reducing the risk of recurrence and metastasis in breast cancer.In clinical practice,an individualized RNI strategy should be developed based on the patient's residual lymph node tumor load,biological behavior of the tumor,and surgical method.

Objective:To summarize the clinical and genetic characteristics of late-onset facioscapulohumeral muscular dystrophy type 1 (FSHD1) patients, and to compare the differences between late-onset and classic-onset FSHD1 patients.Methods:A retrospective analysis was conducted on the clinical and genetic data of genetically confirmed late-onset FSHD1 patients (age at onset30 years) between January 2007 and June 2024 from the Department of Neurology of Peking University First Hospital and the First Affiliated Hospital of Fujian Medical University. Classic-onset FSHD1 patients (10 yearsage at onset≤30 years) were matched 1∶1 according to sex and disease duration for comparison. The demographic information, the number of D4Z4 repeat units, the distal D4Z4 methylation levels, FSHD Clinical Score (CS), Clinical Severity Score (CSS), and Age-Corrected Clinical Severity Score (ACSS) of these patients were collected. Survival analysis was performed to compare the outcome of lower extremity involvement between late-onset and classic-onset FSHD1 patients. The correlation of the number of D4Z4 repeat units and D4Z4 methylation level with CS and ACSS was analyzed in late-onset FSHD1 patients.Results:A total of 61 patients with late-onset FSHD1 were enrolled, 33 (54.1%) of whom are female, with an age of 54.0 (46.0, 62.0) years and a disease duration of 14.0 (5.5, 22.5) years. Compared to classic-onset FSHD1 patients, late-onset patients exhibited significantly lower CS [7.0 (5.6, 8.4) vs 6.0 (4.4, 7.7), U=1 416.000, P=0.013], CSS [3.0 (2.8, 3.3) vs 3.0 (2.0, 4.0), U=2 352.000, P=0.010], and ACSS [189.2 (137.1, 241.3) vs 96.8 (61.3, 132.2), U=3 225.500, P0.001], and higher proportion of patients with limb girdle involvement but no facial muscle involvement [18.0% (11/61) vs 6.6% (4/61), χ2=3.725, P=0.054]. Kaplan-Meier survival analysis showed that the onset age of lower extremity involvement in late-onset patients (45 years, 95% CI 42-48 years) was significantly higher than that in classic-onset patients (24 years, 95% CI 21-27 years, χ2=61.012, P0.001). The duration from symptom onset to lower extremity involvement in late-onset patients (15 years, 95% CI 10-20 years) was significantly longer than that in classic-onset patients (8 years, 95% CI 3-13 years, χ2=9.105, P=0.003). Late-onset FSHD1 patients carried higher average distal D4Z4 methylation levels compared to those with classic-onset FSHD1 [46.68% (40.79%,52.57%) vs 41.02% (34.03%,48.00%), U=1 378.500, P=0.014]. Among late-onset FSHD1 patients, cytosine-phosphate-guanine 6 (CpG6) methylation levels were significantly negatively correlated with ACSS ( r=-0.278, P=0.025); the number of D4Z4 repeat units were significantly negatively correlated with ACSS ( r=-0.272, P=0.034);CpG6 methylation levels were significantly negatively correlated with CS ( r=-0.441, P=0.003), while no correlation was found between number of D4Z4 repeat units and CS ( r=-0.161, P=0.310). Conclusions:Compared with classic-onset FSHD1 patients, late-onset FSHD1 patients are associated with a higher degree of distal D4Z4 methylation, along with a milder muscle weakness phenotype, slower disease progression and a higher proportion of cases without facial muscle involvement. The age at onset can be used as a marker of the severity and prognosis in FSHD1.

Objective:To develope a titanium specimen with good osteogenic activity through fabrication of a composite hydroxyapatite coating on ordered micro-/nanotextured titanium surface.Methods:An ordered micro-/nanotextured structure was prepared on the surface of titanium (the control), and then hydroxyapatite was deposited on the as-prepared ordered micro-/nanotextured structure by alternative loop immersion method. The ordered micro-/nanotextured structures before and after hydroxyapatite deposition were denoted as HA and MN, respectively. Surface morphology was observed using a scanning electron microscope. Bone marrow mesenchymal stem cells (BMMSC) were seeded on the surface of three different materials. Cell morphology was observed with a scanning electron microscope. Cell adhesion and cell proliferation were evaluated using 4', 6-diamidino-2-phenylindole staining and cell counting kit-8 assay, respectively. Extracellular matrix mineralization and the expression levels of osteogenesis-related genes were evaluated by alizarin red staining and real-time quantitative PCR, respectively. Each group has three samples in every experiment.Results:After alternative loop immersing, the MN's original microholes (20 μm in diameter) were retained, and the uniform petal-like hydroxyapatite was deposited on the MN's original titania nanotubes (70 nm in diameter). Compared with the control, BMMSC on MN and HA elongated further and intersected along the micron structure with noticeable pseudopodia and pseudoplates, and the trend was more pronounced especially on HA. The number of early adherent cells on HA was remarkably larger than that on the control and MN at each time point ( P<0.05). On day 1, the A value of cell proliferation on HA was significantly higher than that on the control and MN ( P<0.05). The A value of cell proliferation on HA was significantly lower than that on the control and MN on day 3 ( P<0.05). On day 7, the A value of cell proliferation on HA was significantly lower than that on MN ( P<0.05), but there was no statistically significant difference in the A value of cell proliferation between HA and the control on day 7 ( P>0.05). The Avalue of extracellular matrix mineralization on HA (0.607±0.011) was significantly higher than that on the control and MN (0.268±0.025 and 0.522±0.022, respectively) ( t=-0.25, P<0.001; t=-0.34, P<0.001). The expression levels of bone related genes on HA were significantly higher than those on the control and MN ( P<0.05). Conclusions:HA could promote the BMMSC adhesion and osteogenic differentiation, support BMMSC proliferation, and demonstrate good osteogenic activity.

Objective:To explore the epidemiology and clinical characteristics of hospitalized patients with acute poisoning at the First Affiliated Hospital of Nanchang University in Jiangxi Province.Methods:Based on the electronic medical records management platform of the First Affiliated Hospital of Nanchang University, a retrospective analysis was conducted on hospitalized patients with acute poisoning based on ICD-10 disease codes from 2014 to 2021. The gender, age, residence, poison type, etiology, route of exposure, illness severity, diagnostic and therapeutic processes, hospital stay duration, and prognosis of patients were collected.Results:A total of 1 953 cases of acute poisoning were included, comprising 871 males (44.6%) and 1 082 females (55.4%). The age was (42.7±20.0) years, with 113 fatalities (5.8%). Urban residents accounted for 706 cases (36.1%) and rural residents for 1 247 cases (63.9%). The highest incidence occurred in the 41-60 age group. Suicide was the primary cause, constituting 1 318 cases (67.5%), followed by accidental ingestion or contact, with overdose being less frequent. Oral poison way was the predominant route (1 681 cases, 86.1%), followed by inhalation and contact. Pesticide poisoning was the predominant type of poisoning in this study, with 847 cases (43.4%), followed by drug poisoning and foodborne poisoning. Pesticide poisoning predominated among rural residents (79.2%), while drug poisoning was more prevalent among urban residents (53.5%).Conclusions:In this study, patients with acute poisoning exhibited predominantly female gender, hailed from rural areas, and primary etiology of poisoning was pesticide ingestion for suicidal intent. Targeted education and control measures should be carried out for the above high-risk areas, populations and poisons.