ObjectiveTo systematically compare the differential regulation of the maternal-fetal interface cell lineages and communication networks in the CBA/J×DBA/2 mouse model of recurrent pregnancy loss （RPL） by the two classic therapeutic methods-tonifying the kidney to stabilize the fetus and invigorating the spleen to stabilize the fetus （Shoutai Wan， Juyuan Jian）-of traditional Chinese medicine （TCM） at the single-cell resolution and clarify their modern scientific connotations. MethodsFemale non-pregnant CBA/J mice were caged with male BALB/c （blank group） and DBA/2 （modeling group） mice separately. Pregnant mice in the modeling group were randomly grouped as follows： high/low-dose Shoutai Wan， high/low-dose Juyuan Jian， model （RPL）， and positive control （dydrogesterone）， with 10 mice in each group. Starting from the day after the detection of the vaginal plug， mice were administrated with drugs or an equal volume of normal saline by gavage for 10 consecutive days. After the intervention， the following indicators were measured. ① Macroscopic evaluation： general conditions， uterine wet weight， embryo loss rate， four coagulation parameters ［prothrombin time （PT）， activated partial thromboplastin time （APTT）， fibrinogen （FIB）， and thrombin time （TT）］， and peripheral blood estradiol （E₂） and progesterone （Pg） levels. The decidua with embryos was stained with hematoxylin-eosin （HE） and evaluated by transmission electron microscopy （TEM）. The expression of B-cell lymphoma-2 （Bcl-2）， vascular endothelial growth factor （VEGF）， angiotensin Ⅱ （AngⅡ）， matrix metalloproteinase-2 （MMP-2）， interleukin-6 （IL-6）， leukemia inhibitory factor （LIF）， CXC chemokine ligand 12 （CXCL12）， and microtubule-associated protein 1 light chain 3 homolog （LC3）Ⅰ/Ⅱ was quantified by Western blot. ② Mechanism analysis at the single-cell level： The decidua with embryos from the blank， model， high-dose Shoutai Wan， and high-dose Juyuan Jian groups （6 mice per group， with 3 single-cell samples per group， totaling 24 mice） were analyzed by the BD Rhapsody™ platform， and the whole-cell atlas was drawn by uniform manifold approximation and projection （UMAP） dimensionality reduction clustering combined with the single-cell mouse cell atlas （scMCA）. The differentially expressed genes （DEGs） and cell interaction networks were analyzed via Gene Ontology （GO）， Kyoto Encyclopedia of Genes and Genomes （KEGG）， and CellChat， and the protein-protein interaction （PPI） map of subtype cells was constructed. The CytoTRACE pseudo-temporal analysis was performed to explore the developmental trajectories of core immune cells （natural killer cells， NK cells） from maternal and fetal sources. Results① Pathological and Western blot results indicated that compared with the blank group， the RPL group showed an increase in the embryo loss rate （P<0.01）， down-regulated expression of Bcl-2， LIF， MMP-2， and Vegf in the decidua with embryos （P<0.05）， up-regulated protein levels of CXCL-12， AngⅡ， and IL-6 （P<0.05）， blocked angiogenesis， apoptosis-inflammation imbalance， and coagulation dysfunction. Both prescriptions dose-dependently reduced the abortion rate and restored the angiogenesis-inflammation balance， and Shoutai pill showed superior performance in restoring the E₂ level to the Pg level （P<0.05）. ② Single-cell transcriptome analysis indicated that compared with the blank group， the RPL group showed differences in multiple key cell populations such as decidual cells， trophoblast cells， endothelial cells， erythroblasts， NK cells， and macrophages at the maternal-fetal interface. Immunity and angiogenesis were the key links in RPL. Compared with the RPL group， high-dose Shoutai Wan reversed the changes of NK cells in the embryonic layer （upregulating the mRNA levels of 17 genes and downregulating the mRNA levels of 29 genes） and macrophages （upregulating the mRNA levels of 117 genes and downregulating the mRNA levels of 53 genes） through the regulation of gene expression. High-dose Shoutai pill regulated the immune cells to affect unfolded proteins， cell adhesion， and programmed cell death， thereby promoting decidualization and angiogenesis and modulating embryo-membrane development. High-dose Juyuan Jian regulated the key subgroups of NK cells （up-regulating the mRNA levels of 9 genes and down-regulating the mRNA levels of 17 genes） and macrophages （up-regulating the mRNA levels of 110 genes and down-regulating the mRNA levels of 81 genes）， which affected decidual inflammation and apoptosis and intervened in glycolysis. ③ The pseudo-temporal analysis and communication network indicated that the communication frequency of the RPL group decreased. High-dose Shoutai Wan restored maternal-fetal tolerance through pathways such as NKG2D， CDH5， GDF， and FASLG. High-dose Juyuan Jian enhanced the IL-6/LIFR/JAK/signal transducer and activator of transcription 3 （STAT3） and desmosome/SEMA6/tumor necrosis factor-like weak inducer of apoptosis （TWEAK） signaling to improve endometrial receptivity. The RPL group showed an increased proportion of toxic dNK7， a decreased proportion of reparative dNK4， and blocked embryo fNK1. High-dose Shoutai Wan down-regulated dNK7 and up-regulated dNK4. High-dose Juyuan Jian inhibited the terminal differentiation of dNK7 and up-regulated LILRB1， thus restoring the balance of cytotoxicity and repair. ConclusionBoth the kidney-tonifying and spleen-invigorating methods are effective in treating RPL. NK and macrophages are the key immune cells in the interaction between the embryo and the membrane. The kidney-tonifying method （Shoutai Wan） has an advantage in regulating the phenotypes of unfolded protein， cell adhesion， and programmed cell death， and shows expression characteristics closer to the physiological state in the regulation of NKG2D and CDH5 signals. The spleen-invigorating method （Juyuan Jian） has an advantage in regulating epithelial-mesenchymal transition （EMT）， angiogenesis， and glycolysis and shows higher communication intensity in the IL-6 and LIFR pathways.

OBJECTIVES: To assess the biomechanical performance of a novel bridging plate for treating Rockwood III acromioclavicular joint dislocation. METHODS: A novel bridging plate structure was designed based on CT data from a patient with Rockwood type III acromioclavicular joint dislocation, and a finite element model of the bridging plate-acromioclavicular joint interaction was constructed. The stress and deformation characteristics and biomechanical compatibility of the plate under post-reduction, normal loading, and impact loading conditions were analyzed to evaluate its fixation mechanism and clinical advantages. RESULTS: The stiffness of the bridging system was 27.78 N/mm, close to that of acromioclavicular joint ligaments (26.05 N/mm) and meeting the requirements for flexible deformation. Under normal loading, the maximum stress in the bridging system was 88.29 MPa to sustain physiological activities; under impact loading, the maximum stress reached 480 MPa, and the cable underwent plastic deformation to dissipate energy and effectively buffer local stress concentrations, thereby reducing the risk of rigid bone fractures. The high-stress regions in the bone primarily occurred at the edges of the C1-C4 screw holes. The maximum bone stress was 0.762 MPa under normal loading and 5.963 MPa under impact loading, accounting for 2.86% and 1.66% of the corresponding bolt stresses, respectively. CONCLUSIONS The novel bridging plate is better adapted to biomechanical characteristics of the acromioclavicular joint compared to traditional internal fixation. This fixation system provides sufficient stability while allowing physiological micromotion to facilitate postoperative rehabilitation. Significant flexible deformation can occur at the connection between the fixation ring and the cable, and brittle materials should not be used in this region. The issue of stress concentration at the C1-C4 screw holes requires special attention in its clinical application.
Acromioclavicular Joint/surgery* ; Humans ; Bone Plates ; Biomechanical Phenomena ; Finite Element Analysis ; Joint Dislocations/surgery* ; Fracture Fixation, Internal/methods*

Objective:To analyze the acupoint selection law of acupuncture for autism spectrum disorder (ASD) using data mining techniques.Methods:Literature related to acupuncture for ASD was retrieved from the CNKI, SinoMed, VIP, Wanfang, and PubMed databases from the establishment of the databases to April 1, 2022, and then a database of acupuncture prescriptions was established. The frequency analysis of acupoint use was performed using Microsoft Excel 2019; the Apriori algorithm was used to analyze the association law of acupoints/acupoint areas; SPSS 26.0 was used to perform intergroup cluster analysis.Results:A total of 97 relevant articles with 97 acupuncture prescriptions and 98 acupoints/acupoint areas were included. The most frequently used acupoint was Shenmen (HT 7). The acupoint area of Jin's three-needle therapy and the Governor Vessel acupoints are commonly used. The most frequently occurring part of the acupoint/acupoint area was the head, and the most commonly used specific acupoint was the rendezvous acupoint. Association rule analysis yielded 40 groups of acupoints/acupoint areas, and the most commonly used combination was Laogong (PC 8) and Shenmen (HT 7). Four categories were extracted among high-frequency acupoints/acupoint areas by cluster analysis.Conclusion:Acupuncture treatment for ASD mainly selects the head acupoints, mainly selecting the acupoint area of Jin's three-needle therapy and the Governor Vessel acupoints, and paying attention to the use of specific acupoints.

Objective To study the comparability of total prostate specific antigen ( tPSA) meas-urement by four domestic chemiluminescence immunoassays ( DCI) and electrochemiluminescence immuno-assay ( ECI) . Methods A total of 45 serum samples that requested tPSA tests were selected. Four DCIs ( Snibe MAGLUMI 4000, Mindray CL-2000i, Autobio A2000, HYBIOME AE180) and ECI ( Roche Cobas e601) were used to measure tPSA. The precisions of the methods were evaluated. The four DCIs were com-pared with Roche ECI respectively, and the comparability of the test results was analyzed. Wilcoxon signed rank test and Spearman correlation analysis were used to analyze the data. Results The precisions of five methods were good. The tPSA levels measured by Roche Cobas e601, Snibe MAGLUMI 4000, Mindray CL-2000i, Autobio A2000, and HYBIOME AE180 were 14.11(9.92, 36.09), 12.00(8.56, 27.23), 12.10 (8. 60, 29.87), 13.35(9.51, 32.85) and 14.50(9.88, 40.06) μg/L, respectively. The correlation coeffi-cients of Roche with Snibe MAGLUMI 4000, Mindray CL-2000i, and Autobio A2000 were 0.992, 0.989, 0. 957 and 0.983, respectively (all P<0.001). Assuming the tPSA medical decision point for regression equation was 4.0μg/L, the proportional biases of Snibe MAGLUMI 4000, Mindray CL-2000i, Autobio A2000, and HYBIOME AE180 compared with Roche were -10. 88％, -18. 07％, 0. 23％ and 22. 31％, respectively. Conclusion The comparability of tPSA test results is different between 4 DCIs and Roche ECI, which pro-vides some references for clinical application and standardization of the DCI test results.

Objective To validate the performance of 4 domestic chemiluminescence immunoassay (CLIA) systems on 8 tumor markers quantitative assay kits. Methods Four domestic CLIA systems were randomly marked as A, B, C, D and 8 tumor markers, including carbohydrate antigen (CA)125, CA15-3, CA19-9, ferritin (Fer), alpha-fetoprotein (AFP), carcinoembryonic antigen (CEA), prostate-specific an-tigen (PSA) and free PSA (fPSA) were determined. According to the standard of Clinical and Laboratory Standards Institute (CLSI), the precision, methodological comparison and analytical measure range of 4 systems were validated. Clinical serum samples were obtained from patients in Suzhou Hospital. According to the CLSI EP9-A3 protocol, imported equipment was used as the reference system. The biases of medical de-cision points were assumed, and Pearson correlation analysis and Spearman correlation analysis were used to analyze the data. Results The precision verification of CA125 and PSA on A, CA125 and AFP on B, CA125, CEA, AFP and PSA on C, and all 8 tumor markers on D could meet the laboratory quality control requirements. The correlations of the test results between A-D and the imported equipment were significant (all P<0.05) with the correlation coefficients 0.79-0.99, 0.47-0.99, 0.90-0.98 and 0.78-1.00, respec-tively, and the number of acceptable tests at the level of medical decision was 5, 2, 5, 4. All tests were certified to meet the analytical measure range validation. Conclusions The detection performance of 4 do-mestic CLIA systems for all 8 tumor markers are different. The performance of domestic CLIA systems should be tested when choosing one that can meet laboratory quality control requirements.

Objective: To establish primary immune thrombocytopenia (ITP) animal model induced by anti-platelet membrane glycoprotein GPⅠbα antibodies AN51 and R300. Methods: Twenty guinea pigs (6-8 week) were divided into 4 groups. Five guinea pigs in each group were intravenously injected with different doses of AN51 (0.05, 0.1, 0.2 μg/g) and 0.2 μg/g IgG as control. The whole blood was collected from inner angular venous plexus. Platelets number was determined by an automated cell counter and Swiss-Jim method. Then, the similar protocol was used to establish ITP nude mice model by intraperitoneal injection of different concentrations of anti-platelet GPⅠbα antibody R300, respectively. Results: ①Five minutes after intravenous injection of AN51 at 0.05, 0.1 and 0.2 μg/g, the platelet counts of guinea pigs reduced about 0-5%, 50%-60% and 70%-80% compared to the control group, respectively. The difference was statistically significant (P<0.01) . ②Six hours after intraperitoneal injection of R300 at 0.05, 0.1, 0.2 μg/g, the platelet counts of nude mice decreased about 20%-30%, 60%-70% and 80%-90% compared to the control group, respectively. The difference was statistically significant (P<0.01) . The nude mice, injected 0.2 μg/g R300 once a day for 2 weeks, showed typical ITP clinical manifestations including large number of petechiaes or ecchymoses on limbs, head and abdomen. Conclusion AN51 at 0.2 μg/g and R300 at 0.2 μg/g could establish stable ITP model in guinea pigs and nude mice respectively.

Objective To compose and evaluate the measurement uncertainty of two kinds of chemiluminescence detection system using different methods.Methods The measurement uncertainty was composed by 4 different methods:(1) U 1％ was composed of within-run CV(CVw ％),between-run CV(CVB ％)and bias (CVBias ％);(2) U2％ was composed of CVB ％ and uncertainty of calibration (CVcal ％);(3) U3％ was composed of CVW％,CVs％ and CVcal％;(4) U4％ was composed of CVW％,CVB％,CVBias％ and CVcal％.The measurement uncertainty of Architect i2000SR system (Abbott,USA) and DXI800 system (Beckman,USA) was assessed.Pearson correlation analysis,Spearman correlation analysis,Paried t test and Mann-Whitney u test were performed to analyze the data.Results For Architect i2000SR system,U1％,U2％,U3％ and U4％ were significantly correlated (r=0.727-0.988,all P＜0.05),U3％ and U2％ were significantly different (t =6.88,P＜0.05),U4％ and U1％ were significantly different (t =6.21,P＜0.05).For DXI800 system,U1％,U2％,U3％ and U4％ were also significantly correlated (r =0.608-0.975,all P＜0.05),no significant difference was found between U3％ and U2％ (z=-1.33,P＞0.05),or between U4％ and U 1％ (z =-1.04,P＞ 0.05);the expanded measurement uncertainty was correlated with CVW％,CVB％,CVBias％(rs=0.653-0.912,all P＜0.05),but not with CVcal％(rs=0.548,P＞0.05).Conclusions For Architect i2000SR system,the fourth method is more proper to compose the measurement uncertainty (U4％).For DXI800 system,the first method is more appropriate (U1％).According to the contribution of different components to the measurement uncertainty,the measurement quality could be improved by reducing the imprecision and bias.

Objective To compare the differences of quality of life ( QOL) between self-rating report and guardian′ report through evaluating and describing the QOL of minors with malignant bone tumors. Methods Totally 68 cases of minors with malignant bone tumors hospitalized in Peking Union Medical College Hospital and their parents were selected from October 2012 to October 2015, and were investigated by PedsQL 4.0 scale and PedsQL 3. 0 scale to collect the QOL of minors evaluated by patients and their parents. Results The overall QOL of minors with malignant bone tumors was at a low level. The results of QOL of parents′evaluation were consistent with the results of self-evaluation of minors ( P>0.05) . However, the scores of self-evaluation in physiology, pain, nausea and vomiting, operational anxiety, anxiety of treatment and worries were higher than those of results of parents′ evaluation (P<0.05), and the score of school function of self-evaluation was lower than that of parents′ evaluation ( P<0. 05 ) . Conclusions As for the QOL that evaluated by minors and parents, when the minors have difficulties in self-evaluation, the parents′ report has some reference value. As a individual, the QOL from minors themselves and from parents may be different, so in clinical work, we should provide comprehensive personized and high-quality nursing based on the fully combination of minors self-evaluation and parents′report.

This study aimed to modify the mixed and purified culture of rat retinal ganglion cells (RGCs) in vitro. The retinae of 1-3 day old Sprague-Dawley (SD) rats were separated bluntly into two layers: inner layer and outer layer, under a surgical microscope. Retinal cells isolated from different layers (inner layer, outer layer and whole retinal tissue) by using enzyme dissociation method were cultured in F12/DMEM medium containing 15% FBS. After 3-day culture, the RGCs in the retinal cells obtained from mixed culture of inner, outer, and whole retinal tissue were identified by immunocytochemical staining of Thy-1.1, and the rate of RGCs to retinal cells (RGCs%) was calculated. Two monoclonal antibodies, anti-macrophages/granulocytes (OX-41) against rat macrophage and antibody against rat Thy-1.1 (OX-7), were used to purify RGCs by either a conventional or modified two-stepped immunopanning procedure (purification in situ). Purified RGCs were seeded at different cell density and cultured in F12/DMEM medium containing 15% FBS. Immunocytochemical staining for Thy-1.1, MTT, and PI-Hoechst33342 fluorescence imaging were used to identify the purity and the viability of RGCs in purified culture of RGCs. The results showed: (1) Immunocytochemistry of different retinal tissue layers culture revealed that the RGCs% was (19.9±1.2)%, (0.5±0.2)%, and (6.2±1.7)% respectively in the mixed culture of inner, outer, and whole retinal tissue, with differences being significant (P<0.05); (2) fluorescent double staining of Hoechst33342 and PI indicated that with the same RGCs%, RGCs obtained from purification in situ grew well with more neurite outgrowth than those by the conventional two-stepped immunopanning method; (3) the viability of purified RGCs seeded at high density was increased and the cells developed complex intercellular networks. The viability of RGCs was declined with the decreasing seeding density, and most cells presented round or oval in shape with thin neurites. It was concluded that: (1) RGCs% in the inner layer retina was higher than that in the outer layer retina; (2) RGCs obtained by in situ purification had more neurite outgrowth and lower mortality than those by conventional two-stepped immunopanning procedure; (3) the viability of purified RGCs could be increased by increasing cell seeding density to some extent.

This study aimed to modify the mixed and purified culture of rat retinal ganglion cells (RGCs) in vitro.The retinae of 1-3 day old Sprague-Dawley (SD) rats were separated bluntly into two layers:inner layer and outer layer,under a surgical microscope.Retinal cells isolated from different layers (inner layer,outer layer and whole retinal tissue) by using enzyme dissociation method were cultured in F12/DMEM medium containing 15％ FBS.After 3-day culture,the RGCs in the retinal cells obtained from mixed culture of inner,outer,and whole retinal tissue were identified by immunocytochemical staining of Thy-1.1,and the rate of RGCs to retinal cells (RGCs％) was calculated.Two monoclonal antibodies,anti-macrophages/granulocytes (OX-41) against rat macrophage and antibody against rat Thy-1.1 (OX-7),were used to purify RGCs by either a conventional or modified two-stepped immunopanning procedure (purification in situ).Purified RGCs were seeded at different cell density and cultured in F12/DMEM medium containing 15％ FBS.Immunocytochemical staining for Thy-1.1,MTT,and PI-Hoechst33342 fluorescence imaging were used to identify the purity and the viability of RGCs in purified culture of RGCs.The results showed:(1) Immunocytochemistry of different retinal tissue layers culture revealed that the RGCs％ was (19.9±1.2)％,(0.5±0.2)％,and (6.2±1.7)％ respectively in the mixed culture of inner,outer,and whole retinal tissue,with differences being significant (P＜0.05); (2)fluorescent double staining of Hoechst33342 and PI indicated that with the same RGCs％,RGCs obtained from purification in situ grew well with more neurite outgrowth than those by the conventional two-stepped immunopanning method; (3) the viability of purified RGCs seeded at high density was increased and the cells developed complex intercellular networks.The viability of RGCs was declined with the decreasing seeding density,and most cells presented round or oval in shape with thin neurites.It was concluded that:(1) RGCs％ in the inner layer retina was higher than that in the outer layer retina;(2) RGCs obtained by in situ purification had more neurite outgrowth and lower mortality than those by conventional two-stepped immunopanning procedure; (3) the viability of purified RGCs could be increased by increasing cell seeding density to some extent.