ObjectiveTo investigate the effect and underlying mechanism of the Wulao Qisun prescription on pathological new bone formation in ankylosing spondylitis （AS）. MethodsSynovial fibroblasts were isolated from the hip joints of AS patients and observed under a microscope to assess cell morphology. The cells were identified using immunofluorescence staining. The isolated AS fibroblasts were divided into blank group， low drug-containing serum group， medium drug-containing serum group， high drug-containing serum group， and positive drug group. After drug intervention， cell proliferation was measured using the cell counting kit-8 （CCK-8） assay to observe fibroblast growth and determine the optimal intervention time. Alkaline phosphatase （ALP） activity was measured using the alkaline phosphatase assay. Protein expression of osteocalcin （OCN）， osteopontin （OPN）， and runt-related transcription factor 2 （Runx2） was detected by Western blot. The mRNA expression levels of Wnt5a， β-catenin， and Dickkopf-1 （DKK-1） were measured by real-time quantitative polymerase chain reaction （Real-time PCR）. ResultsCompared with the blank group， each drug-containing serum group of Wulao Qisun prescription and the positive drug group inhibited the proliferation of AS fibroblasts and reduced ALP expression （P<0.01）. Compared with the blank group， the low drug-containing serum group of Wulao Qisun prescription downregulated β-catenin mRNA expression （P<0.05）. The medium and high drug-containing serum groups and the positive drug group significantly downregulated Wnt5a and β-catenin mRNA expression （P<0.05， P<0.01）， with the positive drug group showing the most pronounced effect （P<0.01）. The high drug-containing serum group and the positive drug group significantly upregulated DKK-1 mRNA expression （P<0.01）. Compared with the blank group， the low drug-containing serum group of Wulao Qisun prescription inhibited the expression of OPN and Runx2 proteins （P<0.05， P<0.01）， while the medium and high drug-containing serum groups and the positive drug group inhibited the expression of OCN， OPN， and Runx2 proteins （P<0.05， P<0.01）. ConclusionThe Wulao Qisun prescription can inhibit the proliferation and osteogenic differentiation of AS fibroblasts， thereby delaying the formation of pathological new bone in AS. The possible mechanism involves the regulation of Wnt/β-catenin-related gene expression， further inhibiting the transcription of downstream target genes.

Objective To explore the the correlation between cystatin C(Cys C),beta-2 microglobulin(β2-MG)and ischemic cerebral small vessel disease(CSVD)and its subgroups.Methods Totally 234 patients with CSVD were assigned to the study group,and 92 elderly people with no abnormal findings in head MRI were selected as controls.The CSVD patients were further divided into the subgroups of lacunar infarction(LI),white matter lesion(WML)and LI+WML.Each group was compared risk factors include the blood level of Cys C and β2-MG.Results There were statistically significant differences between CSVD group and control group in cystatin C(Cys C)and β2-MG(P<0.05).Cystatin C(Cys C)and β2-MG there were statistically significant differences between WML group and control group(P<0.05),and also between WML+LI group and control group(P<0.05).Logistic regression analysis and comparison across subgroups showed Cys C and β2-MG to be the common risk factors for WML group and WML+LI group inpatients with ischemic cerebral small vessel disease.Conclusion Cys C and β2-MG are the common risk factors for WML group and WML+LI group inpatients with ischemic cerebral small vessel disease.The risk factors vary across different CSVD subgroups.

ObjectiveTo investigate the effect and mechanism of Yiqi Wenyang Huwei decoction （YWHD） on airway inflammation in bronchial asthma （BA） rats based on transforming growth factor-β₁ （TGF-β₁）/SMAD family member 3 （Smad3） signaling pathway. MethodSixty male SD rats were randomly divided into a normal group， a model group， a dexamethasone （DEX） group， and low-， medium-， and high-dose YWHD groups， with 10 rats in each group. The BA model was induced by intraperitoneal injection of 1 mL of ovalbumin （OVA）-aluminum hydroxide suspension for sensitization， followed by nebulization with 2% OVA. One hour before daily OVA nebulization， the control group was treated with saline， the DEX group with DEX solution at 0.2 g·L^-1， and the low-， medium-， and high-dose YWHD groups with YWHD at 1， 2， 4 g·mL^-1， respectively. General conditions and lung function were observed. Bronchoalveolar lavage fluid （BALF） and serum were collected to count inflammatory cells in BALF and measure immunoglobulin E （IgE） levels in serum and inflammatory cytokines in BALF using enzyme-linked immunosorbent assay （ELISA）. Pathological changes in lung tissues， collagen deposition， and airway mucus secretion were observed by hematoxylin-eosin （HE）， Masson， and periodic acid-Schiff （PAS） staining. TGF-β₁/Smad3-related mRNA and protein levels in lung tissues were determined by Real-time fluorescent quantitative polymerase chain translation （Real-time PCR） and Western blot analysis. ResultCompared with the normal group， the model group showed increased total airway resistance （RL） and decreased dynamic compliance （Cdyn） （P<0.05， P<0.01）， elevated serum IgE levels， increased inflammatory cell counts， and higher inflammatory cytokine levels in BALF （P<0.01）. Additionally， there was significant inflammatory cell infiltration， collagen deposition， and mucus secretion in lung tissues. The levels of TGF-β₁， α-smooth muscle actin （α-SMA）， and Smad3 phosphorylation in lung tissues were significantly increased （P<0.01）. Compared with the model group， the DEX group and high-dose YWHD group exhibited significantly reduced RL （P<0.01）， improved lung dynamic compliance （P<0.05）， and lower serum IgE levels， inflammatory cell counts， and inflammatory cytokine levels in BALF （P<0.05）. Moreover， these treatments alleviated pathological damage in lung tissues and reduced the levels of TGF-β₁， α-SMA， and Smad3 phosphorylation （P<0.01）. ConclusionYWHD reduces airway inflammation， improves pathological damage， and mitigates airway remodeling in bronchial asthma rats， possibly by downregulating TGF-β₁， α-SMA protein levels， and Smad3 phosphorylation.

Objective:To provide references for the application of finite element model in the study of cervical vertebra by statistically analysing the frequency, numerical value, properties, and boundary setting of the finite element model and the corresponding material features as well as boundary settings in the literature.Methods:The literature on cervical vertebra-related finite element models was collected from CNKI, Wanfang, VIP, PubMed, Web of Science, and Embase databases from January 2013 to December 2023. The quality assessment was followed by manual screening. The data sources, application classification, material properties (Young’s modulus and Poisson’s ratio), and boundary conditions of cervical vertebra, cervical intervertebral, and cervical ligaments were statistically analyzed.Results:A total of 102 papers were included. The finite element models of the cervical vertebra were derived from medical image reconstruction modeling techniques, predominantly CT plain scan and magnetic resonance imaging. Among the 102 cervical vertebra models, the C3-C7 (lower cervical segment) model appeared with the highest frequency (19). The Young’s modulus of the cortical bone, cancellous bone, and posterior structure of cervical vertebrae were set at about 12 000 or 10 000, 440, and 3 600 MPa, respectively, and the Poisson’s ratios were mainly set at about 0.29 or 0.30, 0.29, and 0.29. The Young’s modulus of the cervical intervertebral disc endplate, nucleus pulposus, and annulus fibrosus were concentrated around 500 or 2 000, 1, and 100 MPa, respectively, and the Poisson’s ratios were set at about 0.40, 0.50, and 0.40, respectively. The Young’s modulus of the anterior longitudinal ligament, posterior longitudinal ligament, transverse ligament, ligamentum flavum, interspinous ligament, capsular ligament, and articular cartilage of the cervical spine were set around 30, 20, 20, 6-10, 4-8, 10 or 20, 10 MPa, and the Poisson’s ratios were set at aoubt 0.30, 0.30, 0.30, 0.30, 0.30, 0.40, and 0.30, respectively. The Young’s modulus of the upper cervical interdental ligament, lamina, cruciate ligament, nuchal ligament, and pterygoid ligament were set at about 10, 10, 10 or 20, 20, and 5 MPa, respectively, and the Poisson’s ratios were set at about 0.30. Head weight settings were more common at 50, 74, and 100 N.Conclusions:The finite element model of the cervical vertebra has great value in the study of cervical spondylosis, but further optimization is still needed in the assignment of material properties, mesh division, and model verification to improve the accuracy and clinical applicability of the model.

Objective To explore the relationship between lipoprotein(a)[Lp(a)],fibrinogen(Fib)levels and vascular calcification and severity of autogenous arteriovenous fistula(AVF)in maintenance hemodialysis(MHD)patients.Methods A total of 112 MHD patients who visited Suining Central Hospital from November 2021 to November 2023 were selected as the study subjects.According to the degree of AVF vascular calcification,these patients were divided into a non calcified group(n=45)and a calcified group(n=67),in which the calcified group was divided into a mild calcification group(n=19),a moderate calcification group(n=28)and a severe calcification group(n=20).The general information and Lp(a)and Fib levels between non calcified group and calcified group were compared.Multivariate logistic regression model was used to determine the independent risk predictors of AVF vascular calcification in MHD patients.Clinical data of patients with different degrees of calcification were compared.Generalized mixed effects model was used to analyze the relationship between Lp(a),Fib levels and the degree of AVF vascular calcification in MHD patients.Restricted cubic spline model was used to analyze the dose-response relationship between Lp(a)and Fib and severe vascular calcification in AVF.The interaction between Lp(a)and Fib on the severity of vascular calcification in AVF was analyzed.Results Dialysis time,P,PTH,Scr,Hb,BMP-2,FGF-21,Lp(a)and Fib levels of patients in the calcification group were increased(t=17.420,9.640,4.863,6.646,2.158,12.046,13.290,2.395,6.674,all P<0.05),while Ca level was decreased(t=2.820,P=0.006),the differences were statistically significant,respectively.The results of multivariate analysis showed that dialysis time(OR:3.130,95%CI:1.652～5.931),P(OR:4.760,95%CI:2.103～7.133),PTH(OR:3.314,95%CI:1.062～6.045),Scr(OR:2.288,95%CI:1.168～4.481),Hb(OR:4.616,95%CI:2.384～7.949),BMP-2(OR:5.527,95%CI:2.598～9.212),FGF-21(OR:6.242,95%CI:1.201～11.184),Lp(a)(OR:5.509,95%CI:2.787～10.886)and Fib(OR:6.159,95%CI:2.125～12.140)were all independent risk factors for AVF vascular calcification in MHD patients(all P<0.05).There were statistically significant in dialysis time,Ca,PTH,Scr,Hb,BMP-2,FGF-21,Lp(a)and Fib levels among the three groups(F=2.028～6.324,all P<0.05).Patients in the severe calcification group had higher dialysis time,PTH,Scr,Hb,BMP-2,FGF-21,Lp(a)and Fib levels than those in the mild calcification group(t=2.204～11.064)and the moderate calcification group(t=2.025～3.197),while Ca level was reduced(t=3.121,2.471),with statisitcally significant differences(all P<0.05),respectively.The results of the generalized mixed effects model showed that the levels of Lp(a)and Fib were related to the degree of AVF vascular calcification in MHD patients.The results of the restricted cubic spline model showed a non-linear dose-response relationship between Lp(a)and Fib levels and severe vascular calcification in AVF.There was an interactive effect between serum Lp(a)and Fib levels on the severity vascular calcification in AVF(OR=6.324,2.534,all P<0.05).Conclusion Lp(a)and Fib have a certain correlation with the degree of vascular calcification in patients,which may be influencing factors of autogenous AVF vascular calcification in MHD patients.

Objective:To analyze the proportions of innate lymphoid cells (ILCs) subgroups during the process of Mycobacterium tuberculosis (MTB) infection, and to explore the molecular mechanism regulating the differentiation of ILCs during MTB infection. Methods:From March to October 2022, 31 patients with active pulmonary tuberculosis (ATB) and 17 patients who had recovered from pulmonary tuberculosis were enrolled from Shenzhen Third People′s Hospital. Additionally, 30 healthy controls were recruited from the physical examination department. Peripheral blood mononuclear cells (PBMCs) were extracted from all subjects, and the proportions of ILC, ILC1, ILC2 and ILC3 in lymphocytes of PBMCs from different populations were analyzed using flow cytometry. PBMCs from 18 healthy controls were induced in vitro with MTB H37Rv lysate or live Bacillus Calmette-Guérin (BCG) bacteria, and the differentiation of ILC subgroups was analyzed using flow cytometry. CD14 + cells from PBMCs of 29 healthy controls were isolated using magnetic bead sorting technology, and the cells were divided into three groups, including control group, CD14 - group, and CD14 + complement group. The CD14 + complement group was supplemented with CD14 + cells into CD14 - PBMCs through a Transwell chamber, and induced in vitro with H37Rv lysate. The differentiation of ILC subgroups was analyzed using flow cytometry. Statistical analyses were performed using Mann-Whitney U test, Kruskal-Wallis test, and Wilcoxon signed-rank test. Results:The proportions of ILCs in lymphocytes in healthy controls, ATB and recovered tuberculosis groups showed no statistically significant differences ( H=0.07, P=0.965). The proportion of ILC1 in lymphocytes in the peripheral blood of patients with ATB was lower than that in the healthy control group ( U=271.00), and the proportion of ILC2 was higher than that in the healthy control group ( U=299.00). The proportion of ILC1 in the peripheral blood of recovered tuberculosis patients was lower than that in the healthy control group ( U=123.00), and the proportion of ILC3 in the recovered tuberculosis group was higher than those in ATB and healthy control groups ( U=78.00 and 102.50, respectively). All differences were statistically significant (all P<0.05). Compared with the control group, the proportions of ILC ( W=-116.00 and -145.00, respectively) and ILC2 ( W=-149.00 and -155.00, respectively) in lymphocytes decreased after PBMCs induced by H37Rv lysate or BCG live bacteria (all P<0.05). The proportion of ILC1 showed no significant change after induction by H37Rv lysate ( W=-67.00, P=0.154), but decreased after induction by BCG ( W=-121.00, P=0.007) with statistical significance. There was no significant difference in the proportions of ILC1 in lymphocytes among control group, CD14 - group, and CD14 + complement group before and after induction by H37Rv lysate ( W=-159.00, 43.00 and -37.00, respectively, all P>0.05). The proportions of ILC2 in lymphocytes decreased after induction ( W=-435.00, -383.00 and -405.00, respectively) among the three groups, and the differences were all statistically significant (all P<0.001). The proportions of ILC3 in lymphocytes in the control group and CD14 + complement group decreased ( W=-250.00 and -262.00, respectively), and the differences were statistically significant (all P<0.05), while the proportion of ILC3 in lymphocytes in the CD14 - group did not change before and after induction, and the difference was not statistically significant ( W=-172.00, P=0.051). Conclusions:MTB infection induces an imbalance in the differentiation of ILCs subgroups, and the removal of CD14 + cells inhibits MTB-induced ILC3 differentiation without significantly affecting the differentiation of ILC1 and ILC2.

Objective To investigate the correlation between neuropeptide S receptor(NPSR)single nucleotide polymorphism(SNP)(rs323917,rs323920,rs323922,rs2530547,rs324957)and primary insomnia(PI).Methods A total of 157 patients diagnosed with PI in the outpatient department of Center of Sleep Medicine,Gansu Provincial People's Hospital from December 2016 to May 2019 were selected as PI group,and 133 healthy physical examination subjects during the same period were selected as control group.Venous blood samples were collected and DNA,polymerase chain reaction(PCR)amplification and agarose gel electrophoresis were extracted.rs323917,rs323920,rs323922,rs2530547,rs324957 single nucleotide loci were genotypized by target site sequencing.Meanwhile,standard polysomnosis monitoring was performed to analyze the correlation between gene polymorphism and PI.Results There were no significant differences in the genotype distribution of NPSR SNP sites(rs323917,rs323920,rs323922,rs2530547)and allele frequency and rs324957 allele frequency between two groups(P＞0.05).There was significant difference in genotype distribution of rs324957(P=0.034).There was no significant difference in the frequency of different haplotypes between two groups(P＞0.05).Conclusion The expression of rs324957 SNP genotypes in NPSR may be related to PI,and AG genotype is dominant.

Objective:To prepare a 177Lu labeled human epidermal growth factor receptor 2 (HER2) affibody 177Lu-1, 4, 7-triazacyclononane-1, 4, 7-triacetic acid (NOTA)-maleimide (Mal)-cysteine (Cys)-ZHER 2: 342 ( 177Lu-NOTA-MZHER2 for short), and investigate its labeling process and anti-tumor properties. Methods:Two kinds of buffer systems (sodium acetate buffer system and sodium ascorbate buffer system) were investigated. The effects of pH value, precursor mass and reaction temperature on 177Lu labeling NOTA-MZHER2 were compared to obtain optimal labeling conditions. The radiochemical purity of labeled product was determined by instant thin-layer chromatography (ITLC), and its stabilities in PBS and plasma were observed. Human ovarian cancer cell line SKOV-3 was selected for cell internalization and cytotoxicity test to evaluate cell uptake and killing effect of 177Lu-NOTA-MZHER2. SKOV-3 tumor-bearing mice( n=3) were injected with 177Lu-NOTA-MZHER2, and microSPECT/CT imaging was performed. Another 40 tumor-bearing mice were divided into 22.2 MBq group (tail vein injection with probe of 22.2 MBq), control group (tail vein injection with PBS), low-dose group (tumor injection with probe of 3.7 MBq) and high-dose group (tumor injection with probe of 7.4 MBq). Tumor volume and mass of tumor-bearing mice were monitored after injection, and the anti-tumor effect and toxicity of probe were evaluated. Repeated measurement analysis of variance (Bonferroni method) was used to analyze the data. Results:The optimal labeling condition was 70-80 ℃ for 30 min in the system of sodium acetate buffer solution with pH=4 and precursor mass of 50 μg. Under these conditions, the labeling rate of 177Lu-NOTA-MZHER2 was (99.3±0.4)% and radiochemical purity was >99%. After 12 d in PBS and plasma, the radiochemical purities were (95.0±1.5)% and (95.0±2.1)%. Results of cell experiment showed that the internalization of 177Lu-NOTA-MZHER2 accounted for (29.02±3.50)% of the total uptake, and the survival rate of SKOV-3 cells was (48±6)% with the probe concerntration of 6×10 -3 Bq/L. SPECT imaging showed that 177Lu-NOTA-MZHER2 was still concentrated at the tumor site 96 h after injection with a dose of 18.5 MBq. Relative tumor volume (RTV) of tumor-bearing mice in 22.2 MBq group, high-dose group and low-dose group was significantly different from that in control group ( F=21.75, P<0.001). Twenty days after injection, RTV and relative body mass of the tumor-bearing mice in high-dose group were (140±7)% and (80±9)%, respectively. Compared with control group, high-dose group had obvious anti-tumor effect (both P<0.001). Conclusion:177Lu-NOTA-MZHER2 is successfully prepared, which is simple and efficient, and the probe has good anti-tumor effect.

Objective:To radiolabel hyperbranched polymer (HG)-modified liquid metal nanodroplet (LMND)@HG with 177Lu, and explore the radiotherapy sensitization effect on anti-breast cancer therapy. Methods:The ultrasonication method was used to prepare LMND@HG, and then 177LuCl 3 was mixed with LMND@HG to label 177Lu by alloying reactions. The labeling rate, plasma stability and cytotoxicity of 177Lu-LMND@HG were detected. Xenograft mouse model of breast cancer was constructed, and the tumor inhibition test was performed by an intratumoral injection. The tumor progression was monitored by in vivo imaging system. The mechanism of tumor inhibition was verified by immunohistochemistry and immunofluorescence assays. One-way analysis of variance, repeated measures analysis of variance, and the least significant difference t test were used to analyze the data. Results:177Lu was successfully labeled to LMND@HG with a high labeling efficiency >95%. The product did not require further purification and the plasma radiochemical purity was still higher than 95% after 5 d. The cytotoxicity test showed that a dose of 888 kBq (40 mg/L) 177Lu-LMND@HG had obvious toxicity to 4T1 cells, which was significantly lower than 177LuCl 3 (cell viabilities: (16.48±7.81)% vs (85.77±8.87)%; F=77.81, t=11.73, P<0.001) and LMND@HG ((46.53±5.75)%; t=6.20, P<0.001). The biological distribution results showed that 177Lu-LMND@HG was mainly distributed in tumor tissue 5 d after intratumoral injection. The results of the tumor inhibition experiment showed that 1.48 MBq 177Lu-LMND@HG could significantly inhibit the tumor growth compared with the 177LuCl 3 (tumor volume: (222.66±97.70) vs (789.13±245.04) mm 3;F=18.55, t=4.29, P=0.005). In vivo optical imaging of small animals showed that 1.48 MBq and 3.70 MBq 177Lu-LMND@HG both significantly inhibited the tumor growth. Immunofluorescence and immunohistochemical results showed that 177Lu-LMND@HG caused double-stranded DNA break, and suppressed the tumor growth by inhibiting cell proliferation and angiogenesis. Conclusions:A novel 177Lu-liquid metal-based reactive oxygen species (ROS) radiation sensitizer is successfully prepared in this study. The preparation method is efficient and convenient, and the product has high stability. 177Lu-LMND@HG shows an obvious radiotherapy sensitization effect on breast tumor-bearing mice.

Objective:To analyze the proportions of innate lymphoid cells (ILCs) subgroups during the process of Mycobacterium tuberculosis (MTB) infection, and to explore the molecular mechanism regulating the differentiation of ILCs during MTB infection. Methods:From March to October 2022, 31 patients with active pulmonary tuberculosis (ATB) and 17 patients who had recovered from pulmonary tuberculosis were enrolled from Shenzhen Third People′s Hospital. Additionally, 30 healthy controls were recruited from the physical examination department. Peripheral blood mononuclear cells (PBMCs) were extracted from all subjects, and the proportions of ILC, ILC1, ILC2 and ILC3 in lymphocytes of PBMCs from different populations were analyzed using flow cytometry. PBMCs from 18 healthy controls were induced in vitro with MTB H37Rv lysate or live Bacillus Calmette-Guérin (BCG) bacteria, and the differentiation of ILC subgroups was analyzed using flow cytometry. CD14 + cells from PBMCs of 29 healthy controls were isolated using magnetic bead sorting technology, and the cells were divided into three groups, including control group, CD14 - group, and CD14 + complement group. The CD14 + complement group was supplemented with CD14 + cells into CD14 - PBMCs through a Transwell chamber, and induced in vitro with H37Rv lysate. The differentiation of ILC subgroups was analyzed using flow cytometry. Statistical analyses were performed using Mann-Whitney U test, Kruskal-Wallis test, and Wilcoxon signed-rank test. Results:The proportions of ILCs in lymphocytes in healthy controls, ATB and recovered tuberculosis groups showed no statistically significant differences ( H=0.07, P=0.965). The proportion of ILC1 in lymphocytes in the peripheral blood of patients with ATB was lower than that in the healthy control group ( U=271.00), and the proportion of ILC2 was higher than that in the healthy control group ( U=299.00). The proportion of ILC1 in the peripheral blood of recovered tuberculosis patients was lower than that in the healthy control group ( U=123.00), and the proportion of ILC3 in the recovered tuberculosis group was higher than those in ATB and healthy control groups ( U=78.00 and 102.50, respectively). All differences were statistically significant (all P<0.05). Compared with the control group, the proportions of ILC ( W=-116.00 and -145.00, respectively) and ILC2 ( W=-149.00 and -155.00, respectively) in lymphocytes decreased after PBMCs induced by H37Rv lysate or BCG live bacteria (all P<0.05). The proportion of ILC1 showed no significant change after induction by H37Rv lysate ( W=-67.00, P=0.154), but decreased after induction by BCG ( W=-121.00, P=0.007) with statistical significance. There was no significant difference in the proportions of ILC1 in lymphocytes among control group, CD14 - group, and CD14 + complement group before and after induction by H37Rv lysate ( W=-159.00, 43.00 and -37.00, respectively, all P>0.05). The proportions of ILC2 in lymphocytes decreased after induction ( W=-435.00, -383.00 and -405.00, respectively) among the three groups, and the differences were all statistically significant (all P<0.001). The proportions of ILC3 in lymphocytes in the control group and CD14 + complement group decreased ( W=-250.00 and -262.00, respectively), and the differences were statistically significant (all P<0.05), while the proportion of ILC3 in lymphocytes in the CD14 - group did not change before and after induction, and the difference was not statistically significant ( W=-172.00, P=0.051). Conclusions:MTB infection induces an imbalance in the differentiation of ILCs subgroups, and the removal of CD14 + cells inhibits MTB-induced ILC3 differentiation without significantly affecting the differentiation of ILC1 and ILC2.