Objective:To evaluate the changes of right atrial volume and function in patients with atrial septal defect（ASD）by four-dimensional ultrasound automatic quantitative technology，and to explore the predictive value and clinical significance for ASD with pulmonary arterial hypertension（PAH）.Methods:Sixty-one patients with ASD and 32 healthy volunteers（control group）who attended Central China Fuwai Hospital of Zhengzhou University，Fuwai Central China Cardiovascular Hospital from March 2023 to April 2024 were prospectively collected，and classified ASD patients into ASD without PAH group（non-PAH group， n=30）and ASD with PAH group（PAH group， n=31）according to whether or not they had PAH，and obtained right atrial reserve，ductal，systolic longitudinal and circumferential strains（RASr，RAScd，RASct，RASr-c，RAScd-c，RASct-c），right atrial minimal，maximal，and presystolic volumes（RAVmin，RAVmax，and RAVpreA），and calculated right atrial total，passive，and active ejection fraction（RAEF，RAPEF，RAAEF）. The ultrasound parameters and clinical data were compared between groups. Pearson's linear correlation was applied to analyse the correlation between the parameters（RASr，RAA，PASP，NT-proBNP）and mean pulmonary artery pressure（mPAP）. ROC curves were plotted and the area under the curve（AUC）was calculated to analyse the value of the four-dimensional strain parameters of the right atrium in independently and jointly predicting the combination of ASD with PAH. Results:①Compared with the control group，RAVmin，RAVmax，and RAVpreA were elevated in the non-PAH and PAH groups，and the absolute values of RAPEF and RAScd were decreased，whereas the absolute values of RAEF，RAAEF，RASr，RASct，RASr-c，and RASct-c were elevated in the non-PAH group，and decreased in the PAH group（all P<0.05）；compared with the non-PAH group，only RAVmin，RAVmax，and RAVpreA were elevated in the PAH group，and the rest of the above parameters were reduced，with statistically significant differences（all P<0.05）. ②The right atrial four-dimensional strain parameter RASr had the highest predictive value for ASD combined with PAH，with an AUC of 0.876，a specificity of 83.3%，and a sensitivity of 77.4%，respectively. ③RASr was negatively correlated with mPAP（ r=-0.591， P<0.001）and RAA，PASP and NT-proBNP were positively correlated with mPAP（ r=0.539，0.697，0.616；all P<0.001）. ④The predictive value of RASr combined with RAA，PASP，and NT-proBNP was superior for ASD combined with PAH（AUC=0.933）. Conclusions:Four-dimensional ultrasound automatic quantitative technology can effectively evaluate the changes of right atrial volume and function in patients with ASD. ASD patients without PAH have increased right atrial reserve and pump function，and decreased right atrial pipeline function. The right atrial function of ASD patients with PAH is significantly reduced，and the impairment is more severe. Among the right atrial four-dimensional strain parameters，RASr has the highest efficacy in predicting ASD combined with PAH. Moreover，the combination of RASr with RAA，PASP，and NT-proBNP has the best predictive efficacy for ASD combined with PAH，which holds significant clinical value.

Hemoglobin, the principal protein in red blood cells, is crucial for oxygen transport in the bloodstream. The quantification of hemoglobin concentration is indispensable in medical diagnostics and health management, which encompass the diagnosis of anemia and the screening of various blood disorders. Immunological methods, based on antigen-antibody interactions, are distinguished by their high sensitivity and accuracy. Consequently, it is necessary to develop hemoglobin-specific antibodies characterized by high specificity and affinity to enhance detection accuracy. In this study, we immunized a Bactrian camel (Camelus bactrianus) with human hemoglobin and subsequently constructed a nanobody library. Utilizing a solid-phase screening method, we selected nanobodies and evaluated the binding activity of the screened nanobodies to hemoglobin. Initially, human hemoglobin was used to immunize a Bactrian camel. Following four immunization sessions, blood was withdrawn from the jugular vein, and a nanobody library with a capacity of 2.85×10⁸ colony forming units (CFU) was generated. Subsequently, ten hemoglobin-specific nanobody sequences were identified through three rounds of adsorption-elution-enrichment assays, and these nanobodies were subjected to eukaryotic expression. Finally, enzyme-linked immunosorbent assay and biolayer interferometry were employed to evaluate the stability, binding activity, and specificity of these nanobodies. The results demonstrated that the nanobodies maintained robust binding activity within the temperature range of 20-40 ℃ and exhibited the highest binding activity at pH 7.0. Furthermore, the nanobodies were capable of tolerating a 10% methanol solution. Notably, among the nanobodies tested, VHH-12 displayed the highest binding activity to hemoglobin, with a half maximal effective concentration (EC₅₀) of 10.63 nmol/L and a equilibrium dissociation constant (K_D) of 2.94×10^-7 mol/L. VHH-12 exhibited no cross-reactivity with a panel of eight proteins, such as ovalbumin and bovine serum albumin, while demonstrating partial cross-reactivity with hemoglobin derived from porcine, goat, rabbit, and bovine sources. In this study, a hemoglobin-specific high-affinity nanobody was successfully isolated, demonstrating potential applications in disease diagnosis and health monitoring.
Animals ; Camelus/immunology* ; Single-Domain Antibodies/immunology* ; Hemoglobins/immunology* ; Humans ; Peptide Library

Guillain-Barré syndrome (GBS) is an autoimmune-mediated acute polyradiculoneuropathy. Its key feature is focal segmental demyelination along with inflammatory cell infiltration and axonal damage. In clinical practice, anti-ganglioside antibodies in the body fluid of GBS patients show heterogeneity, and conventional immunotherapy is ineffective in some GBS subtypes. Drawing on treatment options for other neuroimmune diseases, targeted regulation of antibody-complement system has now become an important direction in GBS treatment research. This paper examines the immune response mechanisms involving complement, antibodies and related inflammatory factors, explores new strategy in GBS immunotherapy, so as to provide theoretical basis and research entry points for GBS individualized treatment.

Objective:To evaluate the changes of right atrial volume and function in patients with atrial septal defect（ASD）by four-dimensional ultrasound automatic quantitative technology，and to explore the predictive value and clinical significance for ASD with pulmonary arterial hypertension（PAH）.Methods:Sixty-one patients with ASD and 32 healthy volunteers（control group）who attended Central China Fuwai Hospital of Zhengzhou University，Fuwai Central China Cardiovascular Hospital from March 2023 to April 2024 were prospectively collected，and classified ASD patients into ASD without PAH group（non-PAH group， n=30）and ASD with PAH group（PAH group， n=31）according to whether or not they had PAH，and obtained right atrial reserve，ductal，systolic longitudinal and circumferential strains（RASr，RAScd，RASct，RASr-c，RAScd-c，RASct-c），right atrial minimal，maximal，and presystolic volumes（RAVmin，RAVmax，and RAVpreA），and calculated right atrial total，passive，and active ejection fraction（RAEF，RAPEF，RAAEF）. The ultrasound parameters and clinical data were compared between groups. Pearson's linear correlation was applied to analyse the correlation between the parameters（RASr，RAA，PASP，NT-proBNP）and mean pulmonary artery pressure（mPAP）. ROC curves were plotted and the area under the curve（AUC）was calculated to analyse the value of the four-dimensional strain parameters of the right atrium in independently and jointly predicting the combination of ASD with PAH. Results:①Compared with the control group，RAVmin，RAVmax，and RAVpreA were elevated in the non-PAH and PAH groups，and the absolute values of RAPEF and RAScd were decreased，whereas the absolute values of RAEF，RAAEF，RASr，RASct，RASr-c，and RASct-c were elevated in the non-PAH group，and decreased in the PAH group（all P<0.05）；compared with the non-PAH group，only RAVmin，RAVmax，and RAVpreA were elevated in the PAH group，and the rest of the above parameters were reduced，with statistically significant differences（all P<0.05）. ②The right atrial four-dimensional strain parameter RASr had the highest predictive value for ASD combined with PAH，with an AUC of 0.876，a specificity of 83.3%，and a sensitivity of 77.4%，respectively. ③RASr was negatively correlated with mPAP（ r=-0.591， P<0.001）and RAA，PASP and NT-proBNP were positively correlated with mPAP（ r=0.539，0.697，0.616；all P<0.001）. ④The predictive value of RASr combined with RAA，PASP，and NT-proBNP was superior for ASD combined with PAH（AUC=0.933）. Conclusions:Four-dimensional ultrasound automatic quantitative technology can effectively evaluate the changes of right atrial volume and function in patients with ASD. ASD patients without PAH have increased right atrial reserve and pump function，and decreased right atrial pipeline function. The right atrial function of ASD patients with PAH is significantly reduced，and the impairment is more severe. Among the right atrial four-dimensional strain parameters，RASr has the highest efficacy in predicting ASD combined with PAH. Moreover，the combination of RASr with RAA，PASP，and NT-proBNP has the best predictive efficacy for ASD combined with PAH，which holds significant clinical value.

Guillain-Barré syndrome (GBS) is an autoimmune-mediated acute polyradiculoneuropathy. Its key feature is focal segmental demyelination along with inflammatory cell infiltration and axonal damage. In clinical practice, anti-ganglioside antibodies in the body fluid of GBS patients show heterogeneity, and conventional immunotherapy is ineffective in some GBS subtypes. Drawing on treatment options for other neuroimmune diseases, targeted regulation of antibody-complement system has now become an important direction in GBS treatment research. This paper examines the immune response mechanisms involving complement, antibodies and related inflammatory factors, explores new strategy in GBS immunotherapy, so as to provide theoretical basis and research entry points for GBS individualized treatment.

OBJECTIVE: To observe the effects of acupuncture combined with bloodletting on the expression of inflammatory factors in serum, the morphology of sensitized skin tissue and the mast cell degranulation in urticaria rats, and to explore the potential mechanism of this therapy for urticaria. METHODS: Among 42 SD rats of SPF grade, 6 rats were randomly collected for the preparation of sensitized antiserum; and the rest 36 rats were randomized into a blank group, a model group, a positive drug group, an acupuncture group, a bloodletting group and a combined treatment group (acupuncture + bloodletting), 6 rats in each one. The rat model of urticaria was established by passive cutaneous anaphylaxis. In the positive drug group, loratadine (1 mg•kg^-1) by gavage was administered once a day. In the acupuncture group, 1 h after gavage with 0.9% NaCl (1 mL), acupuncture was delivered at "Baihui" (GV 20), "Zhongwan" (CV 12), and bilateral "Quchi" (LI 11) and "Xuehai" (SP 10) for 15 min, once daily . In the bloodletting group, 1 h after gavage with 0.9% NaCl (1 mL), bloodletting was operated at "Dazhui" (GV 14) and bilateral "Geshu" (BL 17), around 0.1 mL of bleeding volume at each point, once daily. In the combined treatment group, 1 h after gavage with 0.9% NaCl (1 mL), the interventions as the acupuncture group and the bloodletting group were adopted, once daily. All the interventions started on day 6 of modeling, lasting 2 weeks. After intervention completion, antigenic stimulation was performed in the rats of each group. Using ELISA, the levels of serum immunoglobulin E (IgE), tryptase (TPS), interleukin-4 (IL-4), interleukin-5 (IL-5), tumor necrosis factor-α (TNF-α) were detected. The diameter of the blue spots of the sensitized skin on the back was measured with ruler in each rat. The morphology of sensitized skin tissue was observed using HE staining, and the degranulation of mast cells was observed using Toluidine blue staining. RESULTS: Compared with the blank group, in the model group, the levels of serum IgE, TPS, IL-4, IL-5 and TNF-α increased (P<0.01), the diameter of blue spot on the sensitized part of the rat back was larger (P<0.01), the degranulation rate of mast cells was elevated (P<0.01), and there were obvious inflammatory cell infiltration and edema in the dermis of sensitized skin tissue on the rat back. Compared with the model group, the serum levels of IgE, TPS, IL-4, IL-5 and TNF-α were reduced in the positive drug group, the acupuncture group, the bloodletting group and the combined treatment group (P<0.01, P<0.05); skin blue spot diameter was smaller in the positive drug group and the combined treatment group (P<0.05); the degranulation rate of mast cells decreased in the positive drug group, the acupuncture group, the bloodletting group and the combined treatment group (P<0.01); and the dermal edema, inflammatory infiltration were attenuated in the positive drug group, the acupuncture group, the bloodletting group and the combined treatment group. Compared with the acupuncture group and the bloodletting group, the serum levels of IgE, TPS, IL-4, IL-5 and TNF-α, as well as the degranulation rate of mast cells in the sensitized tissue were lower in the positive drug group and the combined treatment group (P<0.05). CONCLUSION Acupuncture combined with bloodletting effectively suppress mast cell degranulation in the sensitized skin tissue on the back of urticaria rats, and ameliorate the histopathological morphology. Its effect mechanism may be related to inhibiting the differentiation and proliferation of helper T cells 2 and regulating the humoral immune response.
Animals ; Rats ; Mast Cells/immunology* ; Acupuncture Therapy ; Rats, Sprague-Dawley ; Male ; Cell Degranulation ; Humans ; Bloodletting ; Urticaria/immunology* ; Female ; Acupuncture Points ; Tumor Necrosis Factor-alpha/blood* ; Interleukin-4/blood* ; Interleukin-5/blood* ; Combined Modality Therapy ; Immunoglobulin E/blood* ; Disease Models, Animal

Objective:To investigate the effect and mechanism of isorhynchophylline (IRN) on angiotensin Ⅱ(Ang Ⅱ)-induced cardiac hypertrophy.Methods:H9c2 cells were co-cultured with Ang Ⅱ and different concentrations of IRN (0, 5, 10, 25, 50 μmol/L). The cell surface area and mRNA levels of cardiac hypertrophy markers atrial natriuretic peptide (ANP), brain natriuretic peptide (BNP) and β-myosin heavy chain (β-MHC) were detected to elucidate the effect of IRN on myocardial hypertrophy and the most effective concentration. H9c2 cells were co-cultured with Ang Ⅱ and IRN (25 μmol/L) at different times (0, 6, 12, 24 h) to elucidate the most effective time of inhibition. The phosphorylation levels of the signaling pathway were detected, and the effects of IRN and Akt inhibitor MK2206 on the phosphorylation levels of the signaling pathway were further explored to elucidate the underlying mechanisms.Results:Compared with the control group, the surface area of H9c2 cells, and the mRNA expression of myocardial hypertrophy markers ANP, BNP and β-MHC were significantly increased (all P<0.05). Pretreated with different concentrations of IRN (5, 10, 25, 50 μmol/L) could inhibit the increase in cell surface area induced by AngⅡ (all P<0.05), especially at the concentration of 25 μmol/ L ( P<0.01). IRN could time-dependently inhibit AngⅡ-induced activation of ANP, BNP, β-MHC mRNA (all P<0.05). AngⅡ caused increased phosphorylation levels of Akt, GSK3β, mTOR and FOXO3a. IRN could block AngⅡ-induced phosphorylation of the Akt signaling pathway. Conclusion:IRN attenuates AngⅡ-induced cardiomyocyte hypertrophy by inhibiting the Akt signaling pathway.

Objective During the rapid expansion of postgraduate education,it is meeting more and more conflicts between enroll-ment scale and cultivation quality.The construction of a cultivation quality assurance system for postgraduates becomes an urgent task in the new situation.Methods The data was analyzed with SPSS 24.0,based on the anonymous review results of postgraduates'disserta-tions by a three-A stomatological hospital during 2022-2023.Results The results showed that the overall evaluation of anonymous dis-sertation opinions in 2022 was better than that in 2023,and the passing rate of anonymous dissertation evaluation of doctoral students was higher than that of master students.Irregular forms of composition,low research workload,and insufficient innovation were found in failed postgraduates'dissertations.Conclusion Strengthening the management of study style education and training process,and ex-ploring cultivating mode of overlapping subjects,are necessary to improve the construction of cultivation quality assurance system of postgraduates.

Transcription, Genetic

The automation of traditional Chinese medicine(TCM)pharmaceuticals has driven the development of process analysis from offline to online.Most of common online process analytical technologies are based on spectroscopy,making the identification and quantification of specific ingredients still a challenge.Herein,we developed a quality control(QC)system for monitoring TCM pharmaceuticals based on paper spray ionization miniature mass spectrometry(mini-MS).It enabled real-time online qualitative and quantitative detection of target ingredients in herbal extracts using mini-MS without chromatographic separation for the first time.Dynamic changes of alkaloids in Aconiti Lateralis Radix Praeparata(Fuzi)during decoction were used as examples,and the scientific principle of Fuzi compatibility was also investigated.Finally,the system was verified to work stably at the hourly level for pilot-scale extraction.This mini-MS based online analytical system is expected to be further developed for QC applications in a wider range of pharmaceutical processes.