Objective: This study employed next-generation sequencing (NGS) for large-scale genotyping of 42 blood group systems in blood donors to evaluate its applicability in multi-system blood group identification and rare blood type repository construction, while exploring the complementary value of third-generation sequencing (TGS) in haplotype phasing and variant capture in regions with insufficient sequencing depth for ABO ambiguous samples. Methods: A total of 562 regular blood donors (median donation frequency 11 times) from a blood center were enrolled. NGS was used for genotyping of 42 blood group systems, with a focused analysis on 12 systems essential for rare blood type repository construction: MNS, Lutheran, Kell, Duffy, Kidd, Diego, Yt, Colton, Gerbich, Ok, H, and I. TGS was employed to resolve haplotype phasing (i. e., determining whether different mutation sites are located on the same chromosome) in 12 ABO samples with discrepancies between forward and reverse typing. Serological and genotyping results were compared for MNS, Duffy, Kidd, Lewis, and ABO systems. Results: NGS-based genotyping revealed that Fy (a-b+) in the Duffy system accounted for 0.87% (4/458), and Di (a+b+) in the Diego system accounted for 10.62% (31/292). The Lutheran, Kell, Yt, Colton, H, Ok, I, and Gerbich systems exhibited near-uniform phenotype distributions. Among the 12 ABO ambiguous samples, NGS yielded multiple possible genotype combinations in 5 cases due to inability to phase alleles, whereas TGS uniquely resolved the genotypes through long-read sequencing. In one case, NGS detected only 2 mutation sites due to insufficient sequencing depth, while TGS identified all 11 sites and assigned the A1 phenotype. Concordance rates between serology and genotyping were: Duffy 96.55% (140/145), ABO 96.43% (513/532), Kidd 94.17% (97/103), MNS 85.07% (57/67), and Lewis 68.18% (75/110). In the MNS system, 7 of 8 samples serologically typed as M+N+ but genotyped as M-N+ carried GYPB variants. Lewis discrepancies predominantly featured genotype Le (a-b+) with serological phenotypes of Le (a+b-), Le (a+b+), or Le (a-b-). The NGS platform completed sequencing of 192 samples within 2 weeks. Conclusion: The tiered genotyping strategy combining NGS, TGS, and serology enables multi-system blood group identification in large-scale blood donor populations. TGS provides complementary value in phasing ambiguous ABO samples and detecting variants in regions with insufficient sequencing depth. This study provides a technical framework and baseline frequency data for the expansion of a local rare blood type repository. However, standardization and cost-effectiveness of this strategy require further validation with expanded sample sizes, and serological confirmation should be retained for secretion status-related systems such as Lewis.

Objective To compare postoperative major adverse events in patients with triple-vessel disease undergoing complete versus incomplete revascularization during coronary artery bypass grafting (CABG). Methods A retrospective analysis was conducted on patient data from Tianjin Chest Hospital who underwent primary isolated CABG surgery between 2019 and 2020. Patients were divided into a complete revascularization group and an incomplete revascularization group based on the revascularization status. Inverse probability of treatment weighting (IPTW) was used for risk adjustment. Results A total of 1 419 patients were included in the study, with 1 086 (76.5%) undergoing complete revascularization. IPTW analysis showed that complete revascularization could reduce the incidence of major adverse cardiovascular and cerebrovascular events [HR=0.596, 95%CI (0.404, 0.880), P=0.010] and angina [HR=0.560, 95%CI (0.377, 0.823), P=0.004]. Conclusion In patients with multivessel coronary artery disease, complete revascularization may be associated with improved patient outcomes.

Objective:To construct a rat model of osteoporosis induced by lead exposure, simulate the effects of lead exposure on the skeletal system of rats, and provide reliable basic data support for subsequent research.Methods:In July 2021, 40 eight-week-old SPF-grade SD rats were selected and randomly divided into 5 experimental groups according to body weight stratification randomization: blank control group, positive control group, low-dose, medium-dose and high-dose groups, with 8 rats in each group. Continuous intragastric administration (1 ml/100 g) for 60 days was performed respectively with ultrapure water, 0.4 g/L prednisone acetate, 2.0 g/L, 4.0 g/L, and 8.0 g/L lead acetate (PbAc). During the experiment, the general condition and weight changes of the rats were recorded in detail. After model establishment, biological samples of rats were collected. The levels of blood lead, serum calcium, phosphorus, tartrate-resistant acid phosphatase (TRAP) and bone alkaline phosphatase (BALP) in rats were determined. Additionally, micro-computed tomography (Micro-CT) was used to perform 3D reconstruction of the rat femurs and examine ultrastructural changes. One-way analysis of variance was used to compare multiple groups of data, and LSD or SNK methods were used for pairwise comparisons between groups.Results:During the experiment, there were no obvious abnormalities in feeding and drinking, behavioral activities, and fur color of rats in each group, and the body weight maintained normal and gentle growth. Compared with the blank control group, the levels of blood lead, serum calcium, phosphorus, TRAP and BALP in each dose group of PbAc were significantly increased ( P<0.05), while the femoral bone mineral density, bone volume/total volume and trabecular thickness were significantly decreased ( P<0.05). Compared with the blank control group, the trabecular separation of the femur in the high-dose group was significantly increased, and the trabecular number was significantly decreased ( P<0.05). Compared with the blank control group, the bone mass loss of the femur in each dose group of PbAc was severe in rats. Conclusion:PbAc can cause osteoporosis in rats, and osteoporosis assessment indicators such as bone resorption and bone formation markers in rats, femoral bone mineral density and its ultrastructure can all evaluate the success of model construction.

Objective:To construct a rat model of osteoporosis induced by lead exposure, simulate the effects of lead exposure on the skeletal system of rats, and provide reliable basic data support for subsequent research.Methods:In July 2021, 40 eight-week-old SPF-grade SD rats were selected and randomly divided into 5 experimental groups according to body weight stratification randomization: blank control group, positive control group, low-dose, medium-dose and high-dose groups, with 8 rats in each group. Continuous intragastric administration (1 ml/100 g) for 60 days was performed respectively with ultrapure water, 0.4 g/L prednisone acetate, 2.0 g/L, 4.0 g/L, and 8.0 g/L lead acetate (PbAc). During the experiment, the general condition and weight changes of the rats were recorded in detail. After model establishment, biological samples of rats were collected. The levels of blood lead, serum calcium, phosphorus, tartrate-resistant acid phosphatase (TRAP) and bone alkaline phosphatase (BALP) in rats were determined. Additionally, micro-computed tomography (Micro-CT) was used to perform 3D reconstruction of the rat femurs and examine ultrastructural changes. One-way analysis of variance was used to compare multiple groups of data, and LSD or SNK methods were used for pairwise comparisons between groups.Results:During the experiment, there were no obvious abnormalities in feeding and drinking, behavioral activities, and fur color of rats in each group, and the body weight maintained normal and gentle growth. Compared with the blank control group, the levels of blood lead, serum calcium, phosphorus, TRAP and BALP in each dose group of PbAc were significantly increased ( P<0.05), while the femoral bone mineral density, bone volume/total volume and trabecular thickness were significantly decreased ( P<0.05). Compared with the blank control group, the trabecular separation of the femur in the high-dose group was significantly increased, and the trabecular number was significantly decreased ( P<0.05). Compared with the blank control group, the bone mass loss of the femur in each dose group of PbAc was severe in rats. Conclusion:PbAc can cause osteoporosis in rats, and osteoporosis assessment indicators such as bone resorption and bone formation markers in rats, femoral bone mineral density and its ultrastructure can all evaluate the success of model construction.

Objective To evaluate the clinical efficacy of kissing-stent implantation in the treatment of chronic iliac-vena cava occlusion.Methods The clinical data of 22 patients with chronic iliac-vena cava occlusion,who received kissing-stent implantation,were retrospectively analyzed.The surgical success rate and the procedure-related complications were recorded,the postoperative 3-,6-and 12-month stent patency rates were calculated,and the postoperative 6-month Villalta score was compared with its preoperative value.Results The technical success rate of kissing-stent implantation was 100％.No procedure-related surgical complications occurred.The postoperative 3-,6-and 12-month stent patency rates were 95.5％,90.9％and 86.1％respectively.The postoperative 6-month Villalta score was(12.14±2.80)points,which was remarkably lower than preoperative(20.91±3.16)points,the difference was statistically significant(P＜0.05).Conclusion The implantation of kissing-stent can successfully reconstruct iliac-vena cava with satisfactory short-term efficacy for chronic iliac-vena cava occlusion.

Objective:To investigate the clinical characteristics and prognosis of patients with cutaneous intravascular large B-cell lymphoma (IVLBCL).Methods:The data of 30 cutaneous IVLBCL published between January 1989 and May 2019 in China were systematically reviewed. The clinical manifestation, biochemical and imaging characteristics and diagnostic features of patients were summarized, and then the survival of different groups was also analyzed.Results:The median onset age was 61.5 years old (25.0-83.0 years old), and there were 22 (73.3%) females. All 30 patients presented with cutaneous lesions. Initial symptoms showed cutaneous lesions in 16 (53.3%) patients; and B symptom, respiratory symptoms or central nervous system (CNS) occurred in 14 (46.7%) patients with late cutaneous lesions. Cutaneous lesions were heterogeneous, and 76.7% (23/30) lesions located in lower abdomen and proximal limbs. And 76.2% (16/21) were positive in image examination, and 78.3% (18/23) had two or more extranodal organs invasion. The median time from onset to visit was 2.5 months (0.4-24.0 months), and clinical misdiagnosis rate was 56.7%(17/30). All IVLBCL patients were confirmed by biopsy, including 6 cases (27.3%, 6/22) of bone marrow involvement, 1 case (3.3%) of hemophagocytic syndrome-associated variant, and 29 cases (96.7%) of classical variant. Finally, 81.8% (18/22) patients received anthracycline-based combined chemotherapy. Compared with non-chemotherapy group, the median OS time of chemotherapy group was prolonged [11.0 months (2.0-60.0 months) vs. 2.0 months (0.7-24.0 months), P = 0.002]. Patients with CNS symptoms had shorter median OS time compared with patients without CNS symptoms [2.0 months (0.7-6.0 months) vs. 11.0 months (1.0-60.0 months), P < 0.01]. The median OS time in the group of cutaneous lesions as initial symptom combined with other symptoms was longer than that in group of late cutaneous lesions and other symptoms as initial symptom [unreached (2.0-60.0 months) vs. 3.0 months (1.5-24.0 months), P = 0.032]. Conclusions:Cutaneous IVLBCL is a rare disease with atypical clinical characteristics in China. Prompt attention and biopsy in time will be helpful for early diagnosis. Accompanied with CNS symptoms suggests poor prognosis; and timely chemotherapy can improve the prognosis of the patients.

Objective:To investigate the treatment, safety and prognosis of advanced non-small cell lung cancer patients with leptomeningeal metastasis and performance status score more than 3.Methods:The clinical data of 6 NSCLC patients with leptomeningeal metastasis admitted to the People's Hospital of Wuhan University from November 2016 to September 2018 were analyzed retrospectively. The curative effect and adverse reactions were observed, and the prognosis was analyzed.Results:There were 5 females and 1 male among 6 patients. The median age was 57 years old (46-74 years old). All 6 patients were diagnosed as stage Ⅳ lung adenocarcinoma. There were 3 patients with epidermal growth factor receptor (EGFR) exon 21 mutation, 2 patients with exon 19 mutation and one with anaplastic lymphoma kinase (ALK) fusion mutation. The time window of leptomeningeal metastasis occurred after the progression of adenocarcinoma of lung: 3 cases was more than 12 months, the other 3 cases was less than 12 months, and the average was 20.3 month. Performance status score was more than 3 when leptomeningeal metastasis occurred. The brain magnetic resonance imaging of 6 patients showed linear enhancement of leptomeningeal, cancer cells were found in cerebrospinal fluid in one case, 4 cases were treated with a combination of bevacizumab and EGFR-tyrosine kinase inhibitor (EGFR-TKI), 1 case was treated with oral administration of EGFR-TKI, 1 case was treated with oral administration of EGFR-TKI combined with temozolomide. The median overall survival (mOS) was 9 months (2-13 months), and the median progression free survival was 6 months (2-11 months).Conclusion:Lung adenocarcinoma may be prone to leptomeningeal metastasis; for NSCLC patients with leptomeningeal metastasis and performance status score more than 3, a combination of EGFR-TKI and bevacizumab has good tolerance, high safety and considerable curative effect.

Objective To summarize the clinical characteristics of hepatic lymphoepithelioma-like carcinoma,and to explore the diagnosis and treatment strategies of hepatic lymphoepithelioma-like carcinoma.Methods A retrospectively analysis on 13 patients with liver lymphoepithelioma-like carcinoma in Union Hospital,Tongji Medical College,Huazhong University of Science and Technology,between March 2005 and May 2019 were carried out.Results 8 cases were male,5 were female,median age was 45years (27 to 68 y).There were 8 cases of intrahepatic cholangiocytic lymphoepithelioma-like carcinoma,4cases of hepatocytic lymphoepithelioma-like carcinoma,and 1 case of mixed hepatocytic and cholangiocytic lymphoepithelioma-like carcinoma.All patients received partial hepatectomy and postoperative comprehensive treatment.The patients were followed from 6 months to 7 years.Only one patient died,and the other patients were all in a tumor-free state.Conclusion Primary hepatic lymphoepithelioma-like carcinoma is a rare liver cancer.It is confirmed mainly by pathological examination and immunohistochemistry.With surgery as the main treatment,prognosis is usually fair.

Objective: To investigate the salvage strategy and efficacy for patients with locally recurrent esophageal squamous cell carcinoma after definitive radiochemotherapy. Methods: A total of 126 patients who met the inclusion criteria were enrolled in this study and divided into the salvage surgery, salvage radiochemotherapy and best supportive care. Results: Fifty-eight of 126 patients received salvage esophagectomy, 52 underwent salvage radiochemotherapy and the remaining 16 patients received best supportive care. The 1-, 3-, 5-year overall survival rates of patients receiving salvage therapy were 51%, 16% and 4% for the three groups, whereas all patients in the best supportive care group died within 12.0 months (P<0.001). The 1-, 3-, 5-year survival rates in the salvage surgery and salvage radiochemotherapy groups were 48%, 20% and 7%, and 51%, 11% and 3%, respectively (P=0.473). Multivariate analysis by Cox proportional hazard model showed that T staging of recurrent tumors and salvage regimen were the independent prognostic factors in patients with locally recurrent esophageal cancer (both P<0.001). Postoperative infection occurred in 16% of the patients in the salvage surgery group, and the incidence of esophagotracheal fistula and mediastinoesophageal fistula was 10% and 6% in the salvage radiochemotherapy group. Conclusions A survival benefit can be elicited by salvage surgery or salvage radiochemotherapy in patients with locally recurrent esophageal cancer after definitive radiochemotherapy. Nevertheless, extensive attention should be paid to the management of postoperative complications in clinical practice.

Objective To observe the effect of continuous pumped dexmedetomidine on hemodynamics, awakening quality and stress response in double -lumen tube extubation patients.Methods 60 patients with thoracic surgery were divided into saline group(group N,n =30)and Dex group(group D,n =30).Both two groups were continuously pumped NS and dexmedetomidine respectively 20min before anesthesia.MAP,HR and SpO2 at different time points during the surgery were recorded.Extracted 5ml arterial blood at the time point of T0 and T3,centrifugated and stored at -80 ℃ for testing renin levels.The extubation time,restlessness incidence,orientation recovery time, extubation quality score were recorded.Results The values of MAP in group D at time points of T0,T1,T2,T3,T4 were (97 ±15)mmHg,(102 ±12)mmHg,(103 ±11)mmHg,(98 ±12)mmHg,(96 ±13)mmHg.The values of HR in group D at different time points were (69 ±9)times/min,(80 ±6)times/min,(90 ±7)times/min,(74 ±5)times/min,(73 ±6)times/min.The values of SpO2 in group D at different time points were (95 ±3)%,(98 ±2)%,(98 ± 2)%,(95 ±3)%,(96 ±2)%.The values of MAP in group N at time points of T0,T1,T2,T3,T4 were (96 ± 15)mmHg,(112 ±16)mmHg,(120 ±12)mmHg,(117 ±14)mmHg,(102 ±15)mmHg.The values of HR in group N at different time points were (68 ±7)times/min,(84 ±8)times/min,(105 ±6)times/min,(89 ±6)times/min, (80 ±7)times/min.The values of SpO2 in group N at different time points were (96 ±2)%,(98 ±2)%,(97 ± 3)%,(93 ±3)%,(92 ±4)%.The values of MAP,HR,SpO2 at different time points in group D were significantly lower than those in group N,the differences were statistically significant (the t values of MAP at different time points were tT1 =2.74,tT2 =5.72,tT3 =25.63,tT4 =1.66,all P <0.05;the t values of HR at different time points were tT1 =2.20,tT2 =8.91,tT3 =10.52,tT4 =4.16,all P <0.05;the t values of SpO2 at different time points were tT1 =0.00,tT2 =1.52,tT3 =2.58,tT4 =4.9,PT3 <0.05,PT4 <0.05).There was significant difference between the two groups in orientation recovery time[(3.3 ±2.2)min vs (5.2 ±2.3)min,P =0.0018].There was significant difference between the two groups in agitation score[(1.3 ±0.9)vs (2.4 ±1.2)].There was significant difference between the two groups in extubation quality score[(2.1 ±1.2)vs (3.2 ±1.4),P =0.0018].There was statistical difference between the two groups in T3 adrenal hormones[(N:120 ±25.3)pmol/L,(D:93.4 ±23.2)pmol/L,P =0.00].The dose of sufentanil (60.5 ±9.2)μg in group N was higher than (40.4 ±10.2)μg in group D (P =0.001).The dose of remifentanil (3.3 ±0.8)mg in group N was higher than (2.4 ±0.9)mg in group D (P =0.001).Conclusion Continuous pumped dexmedetomidine can effectively improve awakening quality of patients with double -lumen tube.