Objective: To investigate the genetic basis of RhD-negative phenotype in the blood donor population of Nantong City. Methods: RHD genotyping was performed on 386 randomly selected RhD-negative donor samples (from a total of 676 RhD-negative donors identified between January 20, 2023, and June 28, 2024) using polymerase chain reaction (PCR), and the inconclusive results were confirmed by nucleotide sequencing. Results: Ten RHD allele types were identified: The complete deletion variant RHD 01N.01 was predominant (64.25%, 248/386); followed by RHD 01EL.01 (19.69%, 76/386). RHD 01N.03, RHD 01N.04, RHD 01N.16 and RHD 01EL.32 were frequently observed., RHD 01EL.02, RHD 01EL.08, RHD 01EL.37 and RHD 01N.25 were rare, and two exon deletion variants remained uncharacterized. The phenotypic distribution of RhD-negative blood donors was ccee (55.44%)>Ccee(31.09%)>ccEe(5.96%)>CCee(5.44%)>CcEe(1.81%)>CcEE(0.26%), and the antigen distribution trend was e(99.74%)>c(94.56%)>C(38.60%)>E(8.03%). A correlation was observed between RHD genotypes and RhCE phenotypes. Conclusion: The Nantong blood donor population exhibits unique RHD gene polymorphisms. Integrating RhCE serological phenotyping with RHD genotyping is essential for ensuring transfusion safety.

Objective: To investigate the genetic basis of RhD-negative phenotype in the blood donor population of Nantong City. Methods: RHD genotyping was performed on 386 randomly selected RhD-negative donor samples (from a total of 676 RhD-negative donors identified between January 20, 2023, and June 28, 2024) using polymerase chain reaction (PCR), and the inconclusive results were confirmed by nucleotide sequencing. Results: Ten RHD allele types were identified: The complete deletion variant RHD 01N.01 was predominant (64.25%, 248/386); followed by RHD 01EL.01 (19.69%, 76/386). RHD 01N.03, RHD 01N.04, RHD 01N.16 and RHD 01EL.32 were frequently observed., RHD 01EL.02, RHD 01EL.08, RHD 01EL.37 and RHD 01N.25 were rare, and two exon deletion variants remained uncharacterized. The phenotypic distribution of RhD-negative blood donors was ccee (55.44%)>Ccee(31.09%)>ccEe(5.96%)>CCee(5.44%)>CcEe(1.81%)>CcEE(0.26%), and the antigen distribution trend was e(99.74%)>c(94.56%)>C(38.60%)>E(8.03%). A correlation was observed between RHD genotypes and RhCE phenotypes. Conclusion: The Nantong blood donor population exhibits unique RHD gene polymorphisms. Integrating RhCE serological phenotyping with RHD genotyping is essential for ensuring transfusion safety.

Objective:To investigate the clinical predictive value of Ki67 proliferation index combined with preoperative serum Ctn for postoperative biochemical cure of medullary thyroid carcinoma (MTC) .Methods:Clinical data were collected from Dec. 2008 to Dec. 2024 from 90 patients with surgically confirmed MTC at China-Japan Union Hospital of Jilin University. The optimal cut-off value for preoperative Ctn prediction of biochemical cure (171.18pg/mL) was determined by the ROC curve; the Ki67 proliferation index cut-off value was adopted from the international MTC grading system standard (5%). Patients were divided into three groups based on the above cutoff values: double-low group (Ki67 <5% and Ctn <171.18pg/mL, n=23), single-high group (Ki67 ≥5% and Ctn <171.18pg/mL or Ki67 <5% and Ctn ≥171.18pg/mL, n=49), and double-high group (Ki67 ≥5% and Ctn ≥171.18pg/mL, n=18). The Kaplan-Meier method (Log-Rank and Trend test) was used to compare the differences in biochemical cure rates between groups, and the Cox proportional risk model was used to analyze the risk factors affecting biochemical cure. Results:The correlation between preoperative Ctn and Ki67 proliferation index was not significant. The three groups differed significantly in gender, tumor distribution, tumor size, vascular invasion, N stage, TNM stage, and biochemical cure ( P<0.05), with the double-high group being significantly associated with larger tumors, later N stage and TNM stage, and lower biochemical cure ( P<0.001). Kaplan-Meier analysis showed that the biochemical cure rate in the double-high, single-high, and double-low groups showed a stepwise improvement.Cox univariate analysis showed that tumor size, N stage, TNM stage, preoperative Ctn, and Ki67 combined with Ctn were risk factors for failure to biochemically cure; multivariate analysis confirmed that the double-high group was an independent risk factor ( P<0.05). In the single-high group, the biochemical cure rate of patients in the low Ki67-high Ctn group was lower than that of the high Ki67-low Ctn group and more malignant. Ki67 had less effect on biochemical cure and disease-free survival at the low Ctn level, and Ki67 was an independent risk factor for failure to biochemically cure at the high Ctn level ( P=0.023) and was significantly associated with disease-free survival ( P=0.004) . Conclusions:Serum Ctn is more sensitive than Ki67 index in predicting biochemical cure after MTC, and the correlation between the two was weak. Ki67 proliferation index alone has limited prognostic value, but combines with preoperative Ctn significantly optimize the prognostic assessment of patients.The role of Ki67 index varied at different Ctn levels.

Calcitonin-negative medullary thyroid carcinoma(CNMTC)is a rare subtype of medullary thyroid carcinoma,characterized by normal serum calcitonin levels,which often leads to misdiagnosis or missed diagnosis.The pathogenesis of CNMTC remains unclear and may involve impaired secretion mechanisms or assay-related false negatives.Diagnostic approaches include ultrasound-guided fine needle aspiration cytology,serum CEA and ProGRP measurements,and RET gene testing.Surgical resection remains the mainstay of treatment,while neoadjuvant therapy may be considered in selected cases.This review summarizes recent advances in the understanding,diagnosis,treatment,and prognosis of CNMTC,aiming to provide clinical guidance for better management of this challenging condition.

Objective:To investigate the clinical predictive value of Ki67 proliferation index combined with preoperative serum Ctn for postoperative biochemical cure of medullary thyroid carcinoma (MTC) .Methods:Clinical data were collected from Dec. 2008 to Dec. 2024 from 90 patients with surgically confirmed MTC at China-Japan Union Hospital of Jilin University. The optimal cut-off value for preoperative Ctn prediction of biochemical cure (171.18pg/mL) was determined by the ROC curve; the Ki67 proliferation index cut-off value was adopted from the international MTC grading system standard (5%). Patients were divided into three groups based on the above cutoff values: double-low group (Ki67 <5% and Ctn <171.18pg/mL, n=23), single-high group (Ki67 ≥5% and Ctn <171.18pg/mL or Ki67 <5% and Ctn ≥171.18pg/mL, n=49), and double-high group (Ki67 ≥5% and Ctn ≥171.18pg/mL, n=18). The Kaplan-Meier method (Log-Rank and Trend test) was used to compare the differences in biochemical cure rates between groups, and the Cox proportional risk model was used to analyze the risk factors affecting biochemical cure. Results:The correlation between preoperative Ctn and Ki67 proliferation index was not significant. The three groups differed significantly in gender, tumor distribution, tumor size, vascular invasion, N stage, TNM stage, and biochemical cure ( P<0.05), with the double-high group being significantly associated with larger tumors, later N stage and TNM stage, and lower biochemical cure ( P<0.001). Kaplan-Meier analysis showed that the biochemical cure rate in the double-high, single-high, and double-low groups showed a stepwise improvement.Cox univariate analysis showed that tumor size, N stage, TNM stage, preoperative Ctn, and Ki67 combined with Ctn were risk factors for failure to biochemically cure; multivariate analysis confirmed that the double-high group was an independent risk factor ( P<0.05). In the single-high group, the biochemical cure rate of patients in the low Ki67-high Ctn group was lower than that of the high Ki67-low Ctn group and more malignant. Ki67 had less effect on biochemical cure and disease-free survival at the low Ctn level, and Ki67 was an independent risk factor for failure to biochemically cure at the high Ctn level ( P=0.023) and was significantly associated with disease-free survival ( P=0.004) . Conclusions:Serum Ctn is more sensitive than Ki67 index in predicting biochemical cure after MTC, and the correlation between the two was weak. Ki67 proliferation index alone has limited prognostic value, but combines with preoperative Ctn significantly optimize the prognostic assessment of patients.The role of Ki67 index varied at different Ctn levels.

Calcitonin-negative medullary thyroid carcinoma(CNMTC)is a rare subtype of medullary thyroid carcinoma,characterized by normal serum calcitonin levels,which often leads to misdiagnosis or missed diagnosis.The pathogenesis of CNMTC remains unclear and may involve impaired secretion mechanisms or assay-related false negatives.Diagnostic approaches include ultrasound-guided fine needle aspiration cytology,serum CEA and ProGRP measurements,and RET gene testing.Surgical resection remains the mainstay of treatment,while neoadjuvant therapy may be considered in selected cases.This review summarizes recent advances in the understanding,diagnosis,treatment,and prognosis of CNMTC,aiming to provide clinical guidance for better management of this challenging condition.

Objective To investigate the relationship between abnormal expression of serum endog-enous Apelin precursor peptide,Apela and sST2 and renal failure in elderly patients with CHF,and explore the predictive value of the two marker in predicting renal function deterioration.Methods A total of 210 elderly CHF patients admitted to our department from April 2022 to April 2024 were recruited,and divided into renal failure group(71 cases)and non-renal failure group(139 cases)according to having renal failure or not.Another 100 volunteers who taking out-patient health examination in our hospital during the same period were enrolled as the healthy control group.The expression levels of sST2 and endogenous Apela in the peripheral blood sam-ples of all subjects were detected by immunofluorescence assay and ELISA.The renal function indicators were measured with automatic biochemical analyzer.Pearson correlation analysis was used to analyze the correlation between endogenous Apela and sST2 expression and renal func-tion.ROC curves were plotted to evaluate the predictive value and AUC value of endogenous Apela and sST2 for renal failure in elderly CHF patients.Results The renal failure group had sig-nificantly higher expression levels of endogenous Apela and sST2,and elevated levels of 24-hour urine protein,creatinine and blood urea nitrogen than the non-renal failure group and healthy con-trol group(P＜0.05),and the expression levels of endogenous Apela and sST2 in peripheral blood samples were obviously higher in the non-renal failure group than the healthy control group(P＜0.05).Pearson correlation analysis showed that the expression levels of endogenous Apela and sST2 in the peripheral blood of elderly patients with CHF and renal failure was positively correla-ted with 24-hour urine protein,creatinine and blood urea nitrogen(r=0.346,r=0.752,r=0.565,P＜0.01;r=0.357,r=0.687,r=0.501,P＜0.01).The AUC value of sST2 in diagnosing renal failure in elderly CHF patients was 0.765(95％CI:0.658-0.874),with a sensitivity of 76.1％and a specificity of 79.9％,and the AUC value of endogenous Apela was 0.686(95％CI:0.563-0.809),with a sensitivity of 64.8％and a specificity of 68.3％.When the two markers combined together,the AUC value was 0.818(95％CI:0.712-0.919),the sensitivity was 88.7％,and the specificity was 69.1％.Conclusion High expression of endogenous Apela and sST2 in peripheral blood is closely associated with renal failure in elderly CHF patients,and the two markers have important value in predicting the severity of renal failure in elderly CHF patients.

Objective To investigate the relationship between abnormal expression of serum endog-enous Apelin precursor peptide,Apela and sST2 and renal failure in elderly patients with CHF,and explore the predictive value of the two marker in predicting renal function deterioration.Methods A total of 210 elderly CHF patients admitted to our department from April 2022 to April 2024 were recruited,and divided into renal failure group(71 cases)and non-renal failure group(139 cases)according to having renal failure or not.Another 100 volunteers who taking out-patient health examination in our hospital during the same period were enrolled as the healthy control group.The expression levels of sST2 and endogenous Apela in the peripheral blood sam-ples of all subjects were detected by immunofluorescence assay and ELISA.The renal function indicators were measured with automatic biochemical analyzer.Pearson correlation analysis was used to analyze the correlation between endogenous Apela and sST2 expression and renal func-tion.ROC curves were plotted to evaluate the predictive value and AUC value of endogenous Apela and sST2 for renal failure in elderly CHF patients.Results The renal failure group had sig-nificantly higher expression levels of endogenous Apela and sST2,and elevated levels of 24-hour urine protein,creatinine and blood urea nitrogen than the non-renal failure group and healthy con-trol group(P＜0.05),and the expression levels of endogenous Apela and sST2 in peripheral blood samples were obviously higher in the non-renal failure group than the healthy control group(P＜0.05).Pearson correlation analysis showed that the expression levels of endogenous Apela and sST2 in the peripheral blood of elderly patients with CHF and renal failure was positively correla-ted with 24-hour urine protein,creatinine and blood urea nitrogen(r=0.346,r=0.752,r=0.565,P＜0.01;r=0.357,r=0.687,r=0.501,P＜0.01).The AUC value of sST2 in diagnosing renal failure in elderly CHF patients was 0.765(95％CI:0.658-0.874),with a sensitivity of 76.1％and a specificity of 79.9％,and the AUC value of endogenous Apela was 0.686(95％CI:0.563-0.809),with a sensitivity of 64.8％and a specificity of 68.3％.When the two markers combined together,the AUC value was 0.818(95％CI:0.712-0.919),the sensitivity was 88.7％,and the specificity was 69.1％.Conclusion High expression of endogenous Apela and sST2 in peripheral blood is closely associated with renal failure in elderly CHF patients,and the two markers have important value in predicting the severity of renal failure in elderly CHF patients.

This paper summarized Professor LI Haisong's clinical experience in treating spermatorrhea based on the view of "spirit controlling essence and qi". It is emphasized that the heart spirit has the function of controlling the essence and qi of the human body， believing the frenetic stirring of heart spirit and the insecurity of essence gate are the core pathogenesis of spermatorrhea， LI advocates to regulate the heart spirit first and take into account of the zang-fu organs， essence and qi simultaneously for the treatment. Treatment should be performed according the clinical syndromes differentiatied. For those with heart spirit failing to nourish syndrome， it is recommended to supplement heart qi， nourish spirit and consolidate essence with self-made Yangxin Mijing Formula （养心秘精方）. In case of heart fire hyperactivity， the method of clearing heart heat and draining fire， calming spirit and consolidating essence should be used， and self-made Xiexin Gujing Formula （泻心固精方） is recommended. For heart-liver qi constraint， it is advised to soothe the liver and calm heart， calm the mind and regulate essence with self-made Jieyu Anshen Tiaojing Formula （解郁安神调精方） which is a modifcation to Chaihu Shugan Powder （柴胡疏肝散）. In terms of deficiency of both heart and liver， the treatment principle is supplementing spleen and nourishing heart， calming the mind and controlling essence， for which self-made Xinpi Tongtiao Shejing Formula （心脾同调摄精方） modified from Guipi Decoction （归脾汤） can be used. For deficiency of both heart and kidney， it is better to nourish the kidney and calm heart， calm the mind and consolidate essence with self-made Xinshen Liangzi Tianjing Formula （心肾两滋填精方） that modified from Shuilu Erxian Elixir （水陆二仙丹） and Wuzi Yanzong Pill （五子衍宗丸）. Prescriptions are used to treat the root by harmonizing the zang-fu organs， nourish the spirit by regulating qi and blood， and calm the mind by taking special medi-cinals， and they should be flexibly modified according to the disease.

Trials within cohorts （TwiCs） are design methods derived from randomized controlled trials （RCTS）. They have been widely used in chronic disease areas such as tumors and cardiovascular diseases. The basis of the TwiCs design is a prospective cohort of specific diseases. When RCTS need to be implemented， some patients meeting the inclusion and exclusion criteria are randomly sampled from the cohort to receive "trial interventions"， while the remaining patients in the cohort who meet the inclusion and exclusion criteria continue to receive conventional treatment as control groups. By comparing the efficacy differences between the intervention measures of the trial group and the control group， the efficacy of intervention measures was evaluated. Within the cohort， the same process could be repeated to carry out multiple RCTS， so as to evaluate different intervention measures or compare the efficacy of different doses or timing of interventions. Compared with classical RCTS， TwiCs make it easier to recruit patients from the cohort and have higher external validity， providing a new research paradigm for improving the efficiency and applicability of RCTS in clinical practice. However， TwiCs may also face the challenge of poor compliance of patients in the cohort. Researchers need to take effective measures to control these patients in the design and operation of TwiCs. This article focused on the methodological key points during the implementation of TwiCs， including multi-stage informed consent （patients are informed of consent at three stages： entering the cohort， entering the trial group， and after the trial）， randomization procedures （only random sampling of patients from the cohort to receive "trial interventions"）， sample size calculation， and statistical analysis methods. The article also compared the differences between TwiCs and traditional RCTS and illustrated TwiCs research design and analysis with examples， so as to provide new research ideas and methods for clinical researchers.