1.Effects and safety of gonadotrophin-releasing hormone agonist combined with estradiol patch and oral medroxyprogesterone acetate on endometriosis
Yiqin WANG ; Shaofen ZHANG ; Xun CHEN ; Jin ZHU ; Keqin HUA ; Weigua HU
Chinese Journal of Obstetrics and Gynecology 2009;44(7):504-508
Objective To evaluate effects and safety of gonadotrophin-releasing hormone agonist (GnRH-a) combined with transdermal estradiol and medroxyprogesterone acetate in the treatment of endometriosis. Methods From January I st, 2007 to July 31 st, 2007, 28 endometriosis patients underwent laparnscopic or transabdominal surgery in Obstetrics and Gynecology Hospital affiliated to Fudan University were randomly divided into group A and group B. 14 patients in group A received 3.6 mg goserelin once every 4 weeks, 12 weeks in all 14 patients in group B received goserelin and added 1/2 piece of half-hydrate estradiol every week and 6 mg oral medroxyprogesterone acetate per day, 12 weeks in all. Serum estradiol (E2 ), follicle stimulating hormone(FSH), bone gla protein levels, visual analogue scale (VAS) of pain, bone mineral density of lumbar spine, vaginal exfoliate cell spurs and the form of Kupperman were compared in patients before and after treatment. Results (1 ) After treatment, the level of FSH and E2levels were (5.0 ± 2. 6 ) U/L and (29 ± 17 ) pmol/L in group A and (3.0 ± 1.5 ) U/L, and (87 ± 53 ) pmol/L in group B, which were significantly lower than those before treatment [FSH (17. 0 ± 12. 2) U/L, and E2 (184 ± 194) pmol/L in group A and FSH :(15.3±13.6)U/L and E2: (281±242) pmol/L in group B, P < 0. 01]. On the seventh day after three-month GnRH-a treatment, it was observed that the level of E2 was higher and FSH was lower in group B than the level of E2 and FSH of group A (P < 0. 01 ). (2 ) After treatment, the basal vaginal exfoliate cell proportion in group A [(66. 2 ± 29. 0) %] was significantly lower than that in group B [(11.8 ± 28. 0) %, P < 0. 01] ; while patients in group A owned a lower proportion of the middle [(29. 1 ± 23.1 ) %], superficial layers [(4. 0 ± 5.5 ) %] and esinophilic cells [(2. 3 ± 2. 6)%]than patients group B [middle layer: (73. 0 ± 25.2)% ; superficial layer: (15. 2 ± 10. 9)% ; esinophilic cells: (10. 8 ± 7.9 ) % ; P < 0. 01]. (3) Before the treatment, patients' VAS scores of total, pelvic pain, dysmenorrheal and dyspareunia were 7.43±3. 20,2. 35 ± 1.82, 4. 93 ± 1.98 and 0. 14±0. 53 in group A and were 7.71±2. 02, 2. 57 ± 1.60, 4. 86 ± 1.56 and 0. 29 ± 1.07 in group B; after treatment, the scores above were changed to 0. 14±0. 36,0. 07±0. 27,0. 07±0. 27and 0 in group A and 0. 36±0. 50, 0. 29±0. 47, 0. 07±0. 27 and 0 in group B, which were all significantly lower than those before treatment separately (P <0. 01 ). When menstruation recovered, the scores were 0. 21±0. 43, 0. 07±0. 27, 0. 14 ± 0. 36, and 0 in group A and 0. 50±0. 65, 0. 29±0. 47, 0. 21±0. 43 and 0 in group B, which were also significantly lower than those before treatment (P < 0.01 ), however, no statistical difference was found between groups at any time spot(P > 0. 05). (4) In group A, the bone density after treatment [(0. 96 ± 0. 06 ) g/cm2] was lower than that before treatment [(0. 99 ± 0. 06 ) g/cm2, P < 0.01 )]. In group B, the index was (0. 98 ± 0. 09) g/cm2, which was lower than that before treatment [(0. 99 ± 0. 10 ) g/cm2, P = 0. 201]. No statistical difference was found between groups(P > 0. 05 ). The bone loss rate were (- 2. 77 ± 1.97 ) % in group A and (- 0. 93 ± 2. 86 ) % in group B (P = 0. 058 ). Before treatment, the bone gla protein was (13±3) μg/L in group A and (13±6) μg/L in group B. After treatment, the bone gla protein levels was (17±6)μg/L in group A, which was higher than that before treatment (P < 0. 01 ), the level was (16±6)μg/L in group B, which was higher than that before treatment, however showed no statistical difference(P =0. 053). No difference was found in bone gla protein before and after treatment between two groups (P>0. 05). (5) The form of Kupperman after treatment were 15±7 in group A and 11±6 in group B, which did not show significant difference (P > 0. 05 ). The incidence of flash and sweat were 93% (13/14)in group A, which was significantly higher than that 57% (8/14) in group B(P <0.01 ). Conclusion The add-back therapy that consists of an estradiol patch and oral medroxyprogesterone acetate is effective and safe treatment for endometriosis.
2.Studv on the role of renin angiotensin system in pathogenesis of nonalcoholic fatty liver disease
Xiaona HU ; Dongmei SHI ; Fang LIU ; Zhanjuan GAO ; Haifen MA ; Yiqin HUANG ; Shuming YIN ; Zhijun BAO
Chinese Journal of Digestion 2012;32(6):395-399
Objective To investigate the role of renin angiotensin system (RAS) in pathogenesis of nonalcoholic fatty liver disease (NAFLD).Methods Twenty-four Wistar rats were evenly divided into control group and model group.The rats of control group were fed with normal diet,and model group were with high-fat diet.Rats were killed at the eighth week and serum liver function,blood lipid,glucose and insulin were tested.The liver tissues were stained with HE and Picro acid-Sirius red for pathological observation.The liver tissue concentration of angiotensin Ⅱ was determined by ELISA method and the expression of TGF-β1 in liver tissue was examined by immunohistochemistry.Results After eight weeks high fat feeding,weight,liver index,liver function,blood lipids and serum insulin of model group were significantly higher than those of control group (weight:(463.50±22.72) g vs.(404.29±10.32) g; liver index:(3.75±0.21) g vs.(2.66±0.15) g; ALT:(79.8±8.6) U/L vs.(58.8±11.6) U/L; AST:(200.01±51.72) U/L vs.(150.30±37.27) U/L; total cholesterol:(3.67±0.48) mmol/L vs.(1.50±0.23) mmol/L; triglycerides:(2.06±0.40) mmol/L vs.(0.71±0.34) mmol/L; insulin:(17.37±2.89) pmol/L vs.(11.08±2.12) pmol/L),and all the differences were statistically significant (P<0.01).The histopathological results of model group indicated liver steatosis,inflammatory reaction in part of lobule and portal area and significant fibrosis in part of liver tissue.The liver tissue angiotonin Ⅱ concentration of model group [(32.80 ± 2.81)pg/ml] was higher than that of control group [(22.83 ± 1.75) pg/ml,t =9.559,P<0.01].The immunohistochemistry results showed that the expression of TGF-β1 of model group was obviously higher than that of control group (Z=-2.540,P =0.011 ).Spearman correlation analysis revealed that the increasing degree of angiotensin Ⅱ concentration was positively correlated with liver steatosis scores (r=0.644,P=0.002) and the expression of TGF-β1 (r=0.470,P=0.037).Conclusion The concentration of angiotensin Ⅱ and TGF-β1 increased in the livers of model rats,which indicated that RAS may participate in the pathogenesis and progress of NAFLD.
3.Exploring the prediction model of chronic renal failure based on serum proteomics
Lei HE ; Yawei CHENG ; Ping LIAO ; Heng HU ; Yaming JIN ; Fufeng LI ; Wenjing WANG ; Peng QIAN ; Yiqin WANG
Basic & Clinical Medicine 2010;30(3):263-267
Objective To Screen serum protein markers related to CRF and establish a diagnosis model,exploring and discussing its significance in serodiagnosis by comparing differences of serum protein spectrum expression between patients with chronic renal failure (CRF) and control group.Methods The trial included 62 CRF patients and 28 control ones.Serum samples were tested by surface enhanced laser desorption/ionization-time of flight-mass spectrometry (SELDI-TOF-MS).The data were analyzed to screen serum proteomic biomarkers.By bioinformatics analysis,decision classification tree models were to be established and tested.Results A total of 19 effective protein peaks were significantly different between CRF and normal control (P<0.001) at m/z range of 1 500 to 30 000,among which 18 showed low expression and 1 showed high expression in CRF.CRF and normal control were obviously different in the clustering;By bioinformatics analysis,a CRF-normal controls of the diagnostic decision tree model was developed,which was 87.8% in with prediction accuracy rate of 87.8% sensitivity of 87.1% and a specificity of 89.3%.Condusion Diagnostic decision tree model provides a more accurate prediction and solid experimental evidence for early clinical diagnosis.
4.Effect of cognitive-behavioral therapy on fear of hypoglycemia and self-management behavior in patients with steroid diabetes mellitus
Xiaoqian ZHAO ; Yan HU ; Yiqin PU ; Lei FAN
Chinese Journal of Practical Nursing 2021;37(31):2425-2431
Objective:To explore the effect of cognitive-behavioral therapy on fear of hypoglycemia and self-management behavior of patients with steroid diabetes mellitus.Methods:A total of 76 patients with steroid diabetes mellitus from August 2018 to October 2019 in the First Affilated Hospital of Nanjing Medical University were selected and divided into experimental group and control group according to the admission time, and each group contained 38 cases. The control group recieved routine therapy and nursing care, on the basis of this, the experimental group was given cognitive-behavioral therapy for 8 weeks. Before and after 8 weeks of intervention, the effects were assessed by Chinese version Hypoglycemia Fear Surgey-Worry Scale(CHFSⅡ-WS) and Sumary of Diabetes Self Care Activities (SDSCA), respectively.Results:There was no significant difference in the score of CHFS Ⅱ-WS, SDSCA before intervention between the two groups( P>0.05). After 8 weeks of intervention, the scores of worry & fear and total CHFS Ⅱ-WS scores were (6.61±1.66), (8.75±1.73) points in the experimental group and (7.60±1.44), (9.69±1.77) points in the control group, the differences were statistically significant ( t values were 2.678, 2.247, P<0.05); in addition, the scores of diet, exercise, blood sugar testing, medications in SDSCA were (20.36±1.91), (11.86±2.27), (7.19±1.56), (6.86±0.99) points in the experimental group, and (18.51±1.50), (9.89±2.29), (5.71±1.45), (5.31±1.18) points in the control group, the differences were statistically significant ( t values were 3.644-5.983, P<0.05). Conclusions:Cognitive-behavioral therapy can effectively alliviate fear of hypoglycemia and promote self-management behavior of steroid diabetes mellitus patients.
5.Construction of a quality evaluation index system for specialty care of knee ligament injuries based on a three-dimensional quality structure model
Yiqin YANG ; Sanlian HU ; Xiaomei WEI ; Dan HE ; Weihua HOU ; Haiyan WANG ; Tangyu CHEN ; Qi LIANG
Chinese Journal of Practical Nursing 2024;40(9):692-700
Objective:To construct a nursing quality evaluation index system for knee ligament injury to provide a basis for standardizing the nursing practice and improving the nursing quality of knee ligament injury.Methods:Based on the three-dimensional quality structure model of "structure-process-outcome" proposed by Donabedian, the quality evaluation index system for knee ligament injury specialties was constructed through literature review, brainstorming, and Delphi expert consultation from April to June 2023.Results:Sixteen experts were included in the inquiry. The effective recovery rate of the two rounds of expert correspondence questionnaires was 16/16, the expert authority coefficient was 0.95, and the Kendell harmony coefficients of the expert correspondence were 0.116 and 0.122, respectively (both P<0.05). The final constructed knee ligament injury specialty care quality evaluation index system contained 3 primary indicators (structural quality, process quality and outcome quality), 16 secondary indicators, and 69 tertiary indicators.Conclusions:The specialized nursing quality evaluation index system for knee ligament injury constructed in this study is scientific and reliable, which can provide a basis for the evaluation and assessment of the nursing quality of knee ligament injury specialties and promote the continuous improvement of their nursing quality.
6.Hei Xiaoyaosan Regulates Fas/FasL/Caspase-8/Caspase-3 Signaling Pathway to Inhibit Neuronal Apoptosis in AD Rats
Huping WANG ; Yiqin CHEN ; Jiao YANG ; Yunyun HU ; Yujie LYU ; Zhipeng MENG
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(17):18-26
ObjectiveTo explore the effect and mechanism of Hei Xiaoyaosan in regulating the tumor necrosis factor receptor superfamily member 6 (Fas)/Fas ligand (FasL)/cysteine protease-8 (Caspase-8)/cysteine protease-3 (Caspase-3) signaling pathway to intervene in neuronal apoptosis and prevent Alzheimer's disease (AD). MethodNinety SPF-grade SD male rats of 4 months old were selected and randomly grouped as follows: 10 rats in the blank group, 10 rats in the sham group (bilateral hippocampus injected with 1 μL normal saline), and 70 rats in the modeling group [bilater hippocampus injected with 1 μL amyloid-beta protein 1-42 (Aβ1-42) solution for the modeling of AD]. Fifty successfully modeled rats were selected and randomly assigned into model, donepezil hydrochloride (0.45 mg·kg-1), and high-, medium-, and low-dose (15.30, 7.65, 3.82 g·kg-1) Hei Xiaoyaosan groups. Rats were administrated with corresponding agents by gavage once a day for 42 days. Terminal-deoxynucleoitidyl transferase-mediated nick end labeling (TUNEL) was employed to observe the apoptosis of neurons in the cortex and hippocampus, and immunohistochemistry (IHC) was used to detect the expression of B-cell lymphoma-2 (Bcl-2) and Bcl-2-associated X protein (Bax) in the hippocampus. Real-time fluorescence quantitative polymerase chain reaction(Real-time PCR) was employed to determine the mRNA levels of Fas, FasL, and Fas-associated protein with death domain (Fadd). Western blot was used to determine the protein levels of Fas, FasL, Fadd, Caspase-3, cleved Caspase-3, Caspase-8, and cleved Caspase-8. ResultCompared with the blank group and sham group, the model group showed increased apoptosis rate in the cortex and hippocampus (P<0.01), elevated Bax level (P<0.01), lowered Bcl-2 level (P<0.01), up-regulated mRNA levels of Fas, FasL, and Fadd in the hippocampus (P<0.01), and up-regulated protein levels of Fas, FasL, Fadd, cleaved Caspase-3, and cleaved Caspase-8 (P<0.01). Compared with the model group, donepezil hydrochloride and Hei Xiaoyaosan at high and medium doses decreased the apoptosis rate in the cortex and hippocampus (P<0.05, P<0.01), lowered the Bax level (P<0.01), elevated the Bcl-2 level (P<0.01), and down-regulated the mRNA levels of Fas, FasL, and Fadd and the protein levels of Fas, FasL, Fadd, cleaved Caspase-3, and cleaved Caspase-8 (P<0.05, P<0.01) in the hippocampus. Low-dose Hei Xiaoyaosan decreased the apoptosis rate in the cortex and hippocampus (P<0.05, P<0.01), lowered the Bcl-2 level (P<0.01), and down-regulated the mRNA levels of FasL and Fadd (P<0.05) and the protein levels of Fas, FasL, Fadd, cleaved Caspase-3, and cleaved Caspase-8 (P<0.05) in the hippocampus. ConclusionHei Xiaoyaosan can protect neurons in the cortex and hippocampus of AD rats by inhibiting the apoptosis mediated by the Fas/FasL/Caspase-8/Caspase-3 signaling pathway.
7.Hei Xiaoyaosan Affects Synaptic Plasticity of Hippocampal Neurons in APP/PS1 Double Transgenic Mice of AD via cAMP/PKA/NMDAR Signaling Pathway
Yunyun HU ; Yujie LYU ; Zhipeng MENG ; Jiao YANG ; Yiqin CHEN ; Huping WANG
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(17):27-34
ObjectiveTo explore the effect and mechanism of Hei Xiaoyaosan in modulating the synaptic plasticity in APP/PS1 mice by regulating the cyclic adenosine monophosphate (cAMP)/protein kinase A (PKA)/N-methyl-D-aspartate receptor (NMDAR) signaling pathway. MethodTwelve 4-month-old male C57BL/6J mice were selected as the blank control group, and 60 4-month-old male APP/PS1 double transgenic mice were randomized into model, KW-6002 (adenosine receptor antagonist, 3 mg·kg-1), and high-, medium-, and low-dose (22.10, 11.05, 5.53 g·kg-1, respectively) Hei Xiaoyaosan groups, with 12 mice in each group. Mice were administrated with corresponding drugs for 90 days. Transmission electron microscopy was employed to observe the synaptic ultrastructure of hippocampal neurons, and Golgi staining was used to observe the dendritic spine density of neurons in hippocampal CA1 region. Western blot was employed to measure the protein levels of cAMP, PKA, N-methyl-D-aspartate receptors 1, 2A, and 2B (NR1, NR2A, and NR2B, respectively), postsynaptic density protein 95 (PSD95), and synapsin 1 (SYN1). Real-time fluorescence quantitative polymerase chain reaction(Real-time PCR) was performed to determine the mRNA levels of cAMP, PKA, and NR1. Enzyme-linked immunosorbent assay was employed to determine the content of interleukin-12 (IL-12) and interleukin-4 (IL-4) in the hippocampus. ResultCompared with the blank group, the model group showed blurred boundaries between presynaptic membrane and postsynaptic membrane in hippocampal CA1 region, reduced and scattered synaptic vesicles, and decreased density of postsynaptic membrane, and irregular, disarranged, and loosened dendritic spines of neurons in hippocampal CA1 region (P<0.01). In addition, the model group presented down-regulated protein levels of cAMP, PKA, NR1, NR2A, NR2B, PSD95, and SYN1 and mRNA levels of cAMP, PKA, and NR1, elevated IL-12 level, and lowered IL-4 level in the hippocampus (P<0.01). Compared with the model group, the drug intervention groups showed clear and intact boundaries between presynaptic membrane and postsynaptic membrane in hippocampal CA1 region, increased synaptic vesicles with dense arrangement, increased density of postsynaptic membrane, and improved morphology, arrangement, and density of neuronal dendritic spines (P<0.05, P<0.01). In addition, the drug interventions up-regulated the protein levels of cAMP, PKA, NR1, NR2A, NR2B, PSD95, and SYN1 (P<0.05,P<0.01) and mRNA levels of cAMP, PKA, and NR1 (P<0.01), lowered the IL-12 level (P<0.01), and elevated the IL-4 level (P<0.01) in the hippocampus. ConclusionHei Xiaoyaosan can improve the structure and morphology of hippocampal neurons in APP/PS1 mice by activating the cAMP/PKA/NMDAR signaling pathway and repairing synaptic plasticity.
8.Hei Xiaoyaosan Regulates RAS/RAF/MEK/ERK Signaling Pathway to Ameliorate Oxidative Stress in Rat Model of AD
Huping WANG ; Yujie LYU ; Yunyun HU ; Zhipeng MENG ; Jiao YANG ; Yiqin CHEN
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(17):35-42
ObjectiveTo investigate the role and mechanism of Hei Xiaoyaosan in intervening in oxidative stress in the rat model of Alzheimer's disease (AD) via modulating the rat sarcoma (RAS)/rapidly accelerating fibrosarcoma (RAF)/mitogen-activated protein kinase kinase (MEK)/extracellular signal-regulated kinase (ERK) signaling pathway. MethodOne hundred 4-month-old SPF-grade Wistar male rats were randomly grouped as follows: 10 in the blank group, 10 in the sham group (bilateral hippocampus injected with 1 μL normal saline), and 80 in the modeling group [bilateral hippocampus injected with 1 μL amyloid beta protein 1-42 (Aβ1-42) solution for the modeling of AD]. Fifty rats qualified for modeling were selected and randomized into the model, donepezil hydrochloride (0.5 mg·kg-1), and high-, medium-, and low-dose (15.30, 7.65, 3.82 g·kg-1, respectively) Hei Xiaoyaosan groups. The rats were administrated with corresponding drugs by gavage once a day for 42 consecutive days. At the end of gavage, Morris water maze test was performed to examine the learning and memory abilities of the rats, and Nissl staining was used to observe the pathological changes of neurons in CA3 region of the hippocampus. The immunofluorescence assay was used to observe Aβ deposition and tau phosphorylation. Western blot was employed to determine the protein levels of RAS, RAF, phosphorylated (p)-RAF, MEK, p-MEK, ERK, and p-ERK in the hippocampal tissue. Biochemical methods were used to determine the levels of reactive oxygen species (ROS), malondialdehyde (MDA), and superoxide dismutase (SOD) in the hippocampal tissue. ResultCompared with the sham group, the model group showed prolonged escape latency (P<0.01), shortened swimming distance in the target quadrant (P<0.01), reduced and uneven stained Nissl bodies, enhanced fluorescence intensity of Aβ and p-tau (P<0.01), up-regulated protein levels of RAS, p-RAF, p-MEK, and p-ERK in the hippocampal tissue (P<0.01), increased ROS and MDA content (P<0.01), and decreased SOD activity (P<0.01) on day 5. Compared with the model group, donepezil hydrochloride and high-, medium-, and low-dose Hei Xiaoyaosan shortened the escape latency (P<0.01), increased the swimming distance in the target quadrant (P<0.01), improved the arrangement, morphology, and structures of neurons and the number and distribution of Nissl bodies, decreased the fluorescence intensity of Aβ and p-tau (P<0.01), up-regulated the protein levels of RAS, p-RAF, p-MEK, and p-ERK (P<0.05, P<0.01), decreased the ROS and MDA content (P<0.01), and increased the SOD activity (P<0.01) on day 5. ConclusionHei Xiaoyaosan may ameliorate oxidative stress, reduce Aβ and p-tau levels, and inhibit hippocampal neuronal damage by regulating the RAS/RAF/MEK/ERK signaling pathway, thus improving learning and memory abilities.
9.Hei Xiaoyaosan in Treating Alzheimer's Disease: A Review
Yujie LYU ; Yunyun HU ; Zhipeng MENG ; Yiqin CHEN ; Jiao YANG ; Huping WANG
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(17):43-50
Alzheimer's disease (AD) is a neurodegenerative disease that predominantly affects the elderly. It belongs to the category of dementia in traditional Chinese medicine (TCM), with the onset and progression closely associated with the functions of the kidney, liver, and spleen. The classic TCM formula Hei Xiaoyaosan, which regulates the three Yin of liver, spleen, and kidney, shows broad prospects in treating neurodegenerative diseases. This article reviews the experimental studies reported in the past decade to summarize the mechanisms of Hei Xiaoyaosan and its active components in intervening in AD. Hei Xiaoyaosan can treat AD via multiple targets, levels, and aspects comprehensively. The clinical studies have demonstrated that Hei Xiaoyaosan alone or in combination with other therapies has a definite therapeutic effect on AD. Specifically, it can ameliorate the cognitive impairment, mitigate oxidative stress, and inhibit the overexpression of soluble apoptotic factors in AD patients. This review aims to provide a theoretical basis for the treatment of AD with Hei Xiaoyaosan and explore new research directions. Moreover, it gives new insights into the clinical application of Hei Xiaoyaosan and the development of products with both medicinal and edible values.
10.Analysis and evaluation of hepatitis B test results of blood nucleic acid testing under different screening modes
Yiqin HU ; Jihong HUANG ; Min WANG ; Fangjun FENG ; Jinhui LIU ; Jie DONG
Chinese Journal of Blood Transfusion 2024;37(9):1030-1035
【Objective】 To evaluate the effectiveness of Roche Cobas s 201 in detecting HBV by analyzing its blood nucleic acid testing (NAT) results. 【Methods】 The results were grouped according to the enzyme-linked immunosorbent assay (ELISA) and NAT minipool test (MP), NAT individual test (ID) and repeated NAT ID test (rID), and categorized into 4 groups as ELISA+ /NAT(ID)+ , ELISA+ /NAT(rID)+ , ELISA-/NAT(ID)+ and ELISA-/NAT(rID)+ . The data were statistically analyzed to explore whether there was a difference in the detection of reactive results by repeated NAT, and the correlation between cycle threshold (Ct) and nucleic acid detection rate for NAT-reactive samples with different ELISA results. The true infection status of blood donors was further analyzed by supplementary tests, including NAT systems and chemiluminescence serological marker assays using other methodologies. 【Results】 A total of 1 691 groups of 766 293 blood donor samples were HBV NAT(MP)+ , of which 1 418 groups(83.86%) were detected with reactive results (1 418 HBV NAT+ , 7 090 NAT-), and there were still 273 groups (16.14%) that remained undetected after repeated testing[a total of 1 638 NAT-, Ct(MP): 39.49±3.62]. Of the HBV NAT+ , 881(62.13%) were ELISA+ /NAT(ID)+ , 19(1.34%) were ELISA+ /NAT(rID)+ , 451(31.81%) were ELISA-/NAT(ID)+ , and 67(4.72%) were ELISA-/NAT(rID)+ . For samples with different ELISA results, difference was found in the detection of HBV by repeated NAT (P<0.05). There was no difference in Ct(ID) values between groups ELISA+ /NAT(rID)+ and ELISA-/ NAT(ID)+ , and groups ELISA+ /NAT(rID)+ and ELISA-/ NAT(rID)+ (P>0.05), but there were significant differences between other groups compared pairwise (P<0.05). Supplementary tests were performed on 228 ELISA-/ NAT(MP)+ (ID)- samples, 56 (24.56%) were reactive by chemiluminescent detection of HBsAg+ and 7 (3.07%) by other NAT systems. Among the remaining 221 NAT- samples/donors (96.93%), 53 (23.98%) HBsAg+ donors were likely to have chronic infection, 40 (18.10%) anti-HBe+ and/or anti-HBc+ donors might have previous infections, and the remaining 128 (57.92%) donors who were non-reactive were NAT (MP) pseudo-reactive, with significant differences in anti-HBs levels \'between groups (P<0.05). 【Conclusion】 Repeated NAT has differential detection of donor samples with different reactivity categories or different serologic results, especially within a certain interval, and repeated NAT for ELISA- samples can significantly improve the detection rate. Ct values can assist in assessing the stability and accuracy of the NAT system. For ELISA-/NAT(MP)+ (ID)- donors, the combination of other highly sensitive assays can reduce the risk of viral residuals and safeguard clinical blood safety.