Objective To establish a method for acquisition, perfusion, preservation and transportation of the genetically modified pig kidneys. Methods An eight genetically modified pig was utilized as experimental subject. Prior to kidneys procurement, the health status of the pig was assessed through hematology examination, and the vascular structure of the kidneys was examined using imaging techniques. Following kidneys acquisition, the pig kidneys were perfused and subsequently packaged into the cryogenic storage container labeled "For Organ Transportation Only" for interprovincial transport after communicating the transportation process with transportation department. To evaluate pathological damage to the pig kidneys, a serious of methods were employed such as hematoxylin-eosin (HE) staining, real-time fluorescent quantitative polymerase chain reaction (RT-qPCR), terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) fluorescence staining and enzyme-linked immune absorbent assay (ELISA). Results The preoperative examination of the eight genetically modified pig showed that the serum creatinine was 73.2 μmol/L, blood urea nitrogen was 2.8 mmol/L and hemoglobin was 116 g/L, all within the normal range, indicating normal renal function. CT angiography revealed no lesions in the pig kidneys, and no dilation, stenosis or premature branching of the blood vessels. The total time of obtaining the left and right kidneys from the eight genetically modified pig was (125 ± 10) min, with a blood loss of (20 ± 2) mL. The warm ischemia times were 3 min and 7 min, respectively. The perfusion and trimming times of the left and right kidneys were 36 min and 41 min, respectively. After perfusion, both kidneys were white and moist. The cold preservation and transportation time was 8 h. HE staining showed that some glomeruli were shrunk, and the lumens of the surrounding renal tubules were slightly depressed and swollen with partial inner membrane shedding and microvacuoles formed when the kidneys were preserved for 8 h. The level of cysteinyl aspartate-specific proteinase-3 messenger RNA in the kidneys tissue gradually increased with the extension of cold preservation time after 2 h (P<0.05). TUNEL fluorescence staining showed that only a small number of cells underwent apoptosis after 8 h of cold preservation, which was not significantly different from that at 0 h (P>0.05). ELISA results showed that the contents of lactate dehydrogenase (LDH) and creatinine in the preservation solution remained relatively stable, but the content of kidney injury molecule 1 (KIM-1) gradually increased with the extension of preservation time, suggesting that the pig kidneys had mild injury. Conclusions By establishing methods for acquisition, perfusion, preservation and transportation of the kidneys from genetically modified donor pig, it is possible to effectively and reliably use genetically modified pig kidneys for xenotransplantation.

[Objective] To establish a cryopreservation protocol for small-volume (≤1 mL) red blood cells (RBCs) and to evaluate the reactivity and stability of blood group antigens after cryopreservation, so as to explore its potential application in immunohematology reference laboratories. [Methods] Small-volume RBCs were cryopreserved for 120 days, followed by thawing and deglycerolization to restore the RBC components. The quality of the RBCs was assessed. Serum antibodies were serially diluted and reacted with RBCs before and after cryopreservation, and agglutination scores were recorded to quantitatively evaluate the reactivity and stability of blood group antigens such as Rh, Duffy, Lewis, Kidd, M, and H. Flow cytometry was used to analyze the percentage and mean fluorescence intensity of ABO antigen expression on RBCs before and after cryopreservation to assess the usability of cryopreserved RBCs in flow immunophenotyping and blood group subtype studies. [Results] The hemolysis rate of thawed and deglycerolized RBCs was (0.27±0.10)%, with a supernatant free hemoglobin level of (0.52±0.14) g/L, and the RBC recovery rate was (69.12±7.91)%. The direct antiglobulin test (DAT) was negative for all thawed RBCs. There was no difference in the reactivity of blood group antigens before and after cryopreservation, and no difference in the percentage and mean fluorescence intensity of A and B antigen expression on RBCs before and after cryopreservation. [Conclusion] The small-volume RBC cryopreservation protocol can be applied to immunohematology analysis in reference laboratories and is expected to be widely used in blood group identification, antibody screening, identification, and blood group-related research.

Objective To explore the effect of Huangqi Yanghe Decoction on wound healing of diabetic foot ulcer(DFU)rats based on phosphatidylinositol 3-kinase(PI3K)/protein kinase B(AKT)/nuclear factor-κB(NF-κB)signal pathway.Methods DFU rat model was constructed,and 48 rats successfully modeled were randomly divided into the model group,the Huangqi Yanghe Decoction low(8.5 g/kg)group,the Huangqi Yanghe Decoction high(17 g/kg)dose group and the Huangqi Yanghe Decoction high dose(17 g/kg)+LY294002(PI3K/AKT pathway inhibitor,0.3 mg/kg)group.There were 12 rats in each group.Another 12 rats were selected as the control group.Rats in each group were given corresponding drug intervention for 4 weeks.After the 14th and 28th day-administration,the general state and wound changes of rats were observed,and the wound healing rate was calculated.The fasting blood glucose(FBG)level of rats was measured,and the percutaneous partial pressure of oxygen(TcpO2)of tissue around the wound was detected.Serum levels of vascular endothelial growth factor(VEGF),hypoxia inducible factor-1α(HIF-1α),C-reactive protein(CRP)and interleukin(IL)-6 were determined by enzyme linked immunosorbent assay.Histopathological changes of the wound were observed by hematoxylin-eosin staining.Immunohistochemical staining was used to measure the microvascular density of rat wound tissue.The protein expression levels of PI3K,phosphorylated PI3K(p-PI3K),AKT,phosphorylated AKT(p-AKT),NF-κB p65,phosphorylated NF-κB p65(p-NF-κB p65)and NF-κB inhibitory protein α(IκB-α)in rat wound tissue were determined by Western blot assay.Results Rats in the control group had smooth hair color,normal diet,drinking water and excretion,more active,wound healing fast,less inflammatory reaction in wound tissue,and there were more new blood vessels.Fibroblasts and collagen matrix were abundant in granulation tissue.In the model group,the fur color of rats was dull and matte,and the activity was reduced.The symptoms of polydipsia,polyphagia and polyuria were appeared in the model group,the wound color was dark,and edema and ulcer appeared in the surrounding tissue,a large number of inflammatory cells infiltrated in the wound tissue,accompanied by tissue necrosis and exudation,fewer neovascularization and fibroblasts were observed.Wound healing rate,TcpO2 in wound surrounding tissue,serum VEGF,HIF-1α,microvascular density,p-PI3K,p-AKT and IκB-α protein expression levels in wound tissue were decreased,and FBG,serum CRP,IL-6,p-NF-κB p65 protein expression in wound tissue were increased(P＜0.05).Compared with the model group,the state of rats was gradually improved in the Huangqi Yanghe Decoction low and high dose groups,and the lesion degree of wound tissue was reduced successively,wound healing rate,TcpO2 in wound surrounding tissue,serum VEGF,HIF-1α,microvascular density,p-PI3K,p-AKT and IκB-α protein expression levels in wound tissue were increased in turn(P＜0.05).The FBG,serum CRP,IL-6 and p-NF-κB p65 protein expression in wound tissue were decreased in turn(P＜0.05).LY294002 could partially reverse the therapeutic effect of high-dose Huangqi Yanghe Decoction on DFU rats(P＜0.05).Conclusion Huangqi Yanghe Decoction can regulate PI3K/AKT/NF-κB pathway,inhibit inflammatory response in DFU rats,promote angiogenesis and thus promote wound healing.

Objective To compare the clinical effects of vancomycin, linezolid alone, and their separate combination with spleen polypeptide in the treatment of severe community-acquired pneumonia (CAP) in children with a history of recurrent respiratory infections. Methods A total of 296 children with severe CAP and a history of recurrent respiratory infections were selected as study subjects and randomly divided into four groups: vancomycin group (vancomycin alone), linezolid group (linezolid alone), combined group A (vancomycin combined with spleen polypeptide), and combined group B (linezolid combined with spleen polypeptide), with 74 patients in each group. The curative effects, recovery process (time to normalization of body temperature, time to improvement of cough, time to improvement of respiration, length of hospital stay), adverse reactions, and direct medical costs were compared among the four groups. Decision tree models and cost-effectiveness analysis were used to compare the economics of each treatment plan. Results The total effective rates in the linezolid group, combined group A, and combined group B were 83.78%, 86.49%, and 97.30%, respectively, which were all higher than 71.62% in the vancomycin group, and the combined group B had a higher effective rate than the linezolid group and combined group A (P < 0.05). The time to normalization of body temperature, improvement of respiration, improvement of cough, and length of hospital stay were shorter in the linezolid group, combined group A, and combined group B than in the vancomycin group, and were shorter in the combined group B compared to the linezolid group and combined group A (P < 0.05). There was no statistically significant difference in the occurrence of adverse reactions among the four groups (P>0.05). There were no statistically significant differences in treatment costs, examination/testing costs, and hospitalization costs among the four groups (P>0.05). The treatment costs and total costs were higher in the vancomycin group, combined group A, linezolid group, and combined group B in ascending order, with statistically significant between-group difference (P < 0.05). The cost-effectiveness ratios from high to low were combined group A (75.84), vancomycin group (73.22), linezolid group (32.87), and combined group B (32.69). Sensitivity analysis showed that the cost-effectiveness ratio analysis results were stable and reliable. Conclusion Among the four treatment options, the combination of linezolid and spleen polypeptide has the best therapeutic effect in severe CAP children with a history of recurrent respiratory infections, with the most significant clinical benefit and cost-effectiveness advantages.

Objective To analyze the effects of massage therapy on the inflammatory state and lung function of pediatric refractory Mycoplasma pneumonia (RMPP). Methods A total of 320 children with RMPP treated in Hebei Seventh People′s Hospital from September 2022 to August 2023 were selected in the study. According to different treatment methods, they were divided into two groups, with 160 cases in each group. The western medicine group was given conventional anti-inflammatory and other symptomatic treatment, while the massage group was given dialectical massage treatment based on the western medicine group. The clinical efficacy, duration of disappearance of clinical symptoms, pulmonary function indexes, serum inflammatory factors, immune function indexes and total incidence of adverse reactions were compared between the two groups. Results The total effective rate of massage group was higher than that of control group (P < 0.05). The disappearance time of fever, cough, sputum and lung moist rales in observation group was shorter than that in control group (P < 0.05). After treatment, the peak expiratory flow (PEF), maximum mid-expiratory flow (MMEF) and forced vital capacity (FVC) in the massage group were higher than those in the control group (P < 0.05). The levels of serum C-reactive protein (CRP), tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) in the massage group were lower than those in the control group after treatment (P < 0.05). After treatment, CD3⁺, CD4⁺, CD4⁺/CD8⁺ in the massage group were higher than those in the control group (P < 0.05), and CD8⁺ was lower than that in the control group (P < 0.05). There was nosignificant difference in the total incidence of adverse reactions between the massage group and the western medicine group (P>0.05). Conclusion Massage based on syndrome differentiation combined with western medicine can effectively relieve fever, cough and other symptoms of RMPP children, reduce inflammatory response, improve lung function and immune function, and has less adverse reactions.

Objective To investigate the effects of nicorandil combined with rosuvastatin calcium on monocyte-to-high density lipoprotein cholesterol ratio (MHR), systemic immune-inflammation index (SII), and cardiac function in patients with coronary slow flow (CSF). Methods A group case-control study was used to select 240 patients with CSF confirmed by coronary angiography, and they were randomly divided into observation group and control group, with 120 patients in each group. On the basis of conventional drug treatment, the control group was treated with rosuvastatin calcium, while the observation group was treated with nicorandil combined with rosuvastatin calcium for 6 months. Clinical efficacy, inflammatory markers[high-sensitivity C-reactive protein (hs-CRP), MHR, SII], corrected TIMI frame count (CTFC) of major coronary branches [left anterior descending branch (LAD), left circumflex branch (LCX), right coronary artery (RCA)], cardiac function indicators[left ventricular ejection fraction (LVEF), left ventricular end-diastolic diameter (LVEDD), global longitudinal strain (GLS)], and the occurrence of major adverse cardiovascular events (MACE) were compared between the two groups. Results The total effective rate in the observation groupwas significantly higher than that in the control group (95.00% versus 80.00%, P < 0.05). After treatment, the duration, severity, andfrequency of angina pectoris attacks were reduced or shortened in both groups, with more changes observed in the observation group (P < 0.05). Serum levels of hs-CRP, MHR, and SII were lower in both groups after 6 months of treatment, and the observation group with significantly was lower compared to the control group (P < 0.05). CTFC values of LAD, LCX, and RCA were also lower in both groups after 6 months, with the observation group showing significantly lower values (P < 0.05). The absolute values of LVEF and GLS increased in both groups after 6 months of treatment, and the observation group was higher than the control group (P < 0.05). However, no significant difference was observed in LVEDD and occurrence of MACE between the two groups (P>0.05). Conclusion The application of nicorandil combined with rosuvastatin calcium in the treatment of CSF patients can significantly improve the overall treatment efficacy, reduce hs-CRP, MHR, and SII levels, enhance cardiac function indicators such as LVEF, LVEDD, and GLS, and effectively alleviate the severity of angina pectoris attacks.

【Objective】 To analyze the value of plasmin-α2-plasmin inhibitor complex (PIC) and thrombin-antithrombin complex (TAT) for risk stratification of massive transfusion (MT) in patients with postpartum hemorrhage (PPH). 【Methods】 Clinical data and blood samples of patients with PPH in our hospital from January 2019 to December 2022 were retrospectively analyzed. MT (MT group, n=60) was defined as transfusion of red blood cells≥10 U within 24 h after delivery, and <10 U was defined as non-MT group (n=190). Plasma PIC and TAT levels were detected by chemiluminescence immunoassay at the onset of PPH. 【Results】 Compared with non-MT group, PPH patients in MT group had higher TAT [2.20 (1.20, 3.00) ng/mL vs 4.00 (2.20, 6.30) ng/mL, Z=-5.464, P<0.001] and PIC [0.99 (0.82, 1.13) μg/mL vs 1.11 (1.05, 1.55) μg/mL, Z=-7.228, P<0.001] level. The analysis of receiver operator characteristic curve showed that the area under the curve required for MT after TAT and PIC combined to predict PPH was 0.820 (95% CI: 0.756-0.886), and the positive likelihood ratio was 4.76 and the negative likelihood ratio was 0.35, which was significantly better than the two predictions alone. Multivariate logistics regression analysis showed that TAT level>3.25 ng/mL and PIC level>1.04 μg/mL were independent risk factors for MT after PPH. 【Conclusion】 Elevated TAT and PIC levels are independent predictors of MT in patients with PPH, and their combined predictive efficacy is better.

【Objective】 To estimate the prevalence, associated factors and patterns of multimorbidity of non-communicable diseases among adults in Shaanxi Province so as to provide evidence for the prevention and control of non-communicable diseases. 【Methods】 We used the data of adults aged 18 years and older collected in the baseline survey of Shaanxi Project in the Regional Ethnic Cohort Study in Northwest China. Multinomial logistic regression was used to explore the associated factors for multimorbidity. Exploratory factor analysis was used to extract patterns of multimorbidity. 【Results】 The prevalence of multimorbidity was 10.7% among the 44 442 participants. Age increase, being males, urban residence, and being overweight or obesity were positively associated with multimorbidity. Compared with women, men had a higher risk of multimorbidity. The OR and 95% CI was 1.25 (1.12-1.39). The risk of multimorbidity increased with age among adults. Compared with participants aged 18.0-34.9 years, the ORs and 95% CIs of those aged 35.0-44.9, 45.0-54.9, 55.0-64.9, and ≥65.0 years were 4.73 (3.47-6.46), 15.61 (11.60-21.00), 41.39 (30.76-55.70) and 90.04 (66.58-121.77), respectively. The primary multimorbidity patterns among adults in Shaanxi were cardiovascular-metabolic multimorbidity (5.4%), viscero-articular multimorbidity (1.0%), and respiratory multimorbidity (0.3%). 【Conclusion】 More than one in ten adults in Shaanxi Province had multimorbidity, and the predominant pattern of multimorbidity was cardiovascular-metabolic multimorbidity. The prevention and control of non-communicable diseases should be reinforced in middle-aged and older people, males, people living in the urban, and overweight or obese people. More attention should be paid to the prevention and control of cardiovascular-metabolic diseases.

【Objective】 To evaluate the dietary quality with the dietary balance index (DBI_16) and the association between dietary quality and bone mass among middle-aged and elderly people in Gansu Province so as to provide evidence for improving dietary quality and bone health status of Gansu population. 【Methods】 Based on the information of the type and quantity of food intake and the bone mass of middle-aged and elderly people aged 35 years and above collected by the Gansu Project in the Regional Ethnic Cohort Study in Northwest China, DBI_16 was used to evaluate the intake level of cereals, vegetables, fruits, milk, beans, fish and shrimp, eggs and other foods, and the degree of inadequate, excessive and unbalanced dietary intake of the participants. Multiple linear regression was used to evaluate the associations of three component indexes of DBI_16, high bound score (DBI_HBS), low bound score (DBI_LBS), diet quality distance (DBI_DQD), and seven single indexes of DBI_16 with bone mass. 【Results】 Analyses of the dietary and bone mass data of 11,840 participants showed that 44.8% of participants consumed excessive amounts of cereals compared to the dietary recommendation. 96.3%, 90.6%, 90.1%, 71.9%, 95.1% and 60.3% of participants’ intake of vegetables, fruits, milk, soybeans, fish and shrimp, and eggs, respectively, were inadequate. 47.7% participants consumed less than 10 types of food. 2.3% participants’ DBI_LBS levels were appropriate. 54.7% participants’ DBI_HBS levels were appropriate. Only 1.2% participants’ DBI_DQD reached a balanced level. The bone mass level in the study population was (2.5±0.6) kg [(2.8±0.5) kg for men and (2.3±0.5) kg for women]. After adjusting for sociodemographic characteristics, lifestyle, total dietary energy intake and body mass index, DBI_LBS and DBI_DQD were negatively associated with bone mass [β and 95% CI was -0.002 01 (-0.003 62--0.000 40) and -0.001 76 (-0.003 09--0.000 43), respectively]. 【Conclusion】 Dietary intake imbalance is common among middle-aged and elderly people in Gansu Province, and the more severe the dietary intake imbalance, the lower the bone mass level.

Objective:To investigate the role and mechanism of N 6-methyladenosine (m 6A) methyltransferase-like 3 (METTL3) in vascular calcification (VC) of chronic kidney disease (CKD) through apoptosis-associated protein. Methods:(1) Real-time fluorescence quantitative PCR was used to test METTL3 mRNA in serum of maintenance hemodialysis (MHD) patients. (2) Western blotting was used to detect the expression of METTL3 protein in high-phosphorus stimulated vascular smooth muscle cells (VSMCs), and immunofluorescence double lable was used to observe the distribution of METTL3 and Runt-related transcription factor 2 (Runx2). The METTL3 overexpressed and knockdown plasmids were constructed and transfected into VSMCs. Alizarin red staining was used to detect calcification degree. Western blotting was used to detect the expressions of osteogenic markers [Runx2, bone morphogenetic protein-2(BMP-2), collagen Ⅰ] and apoptosis- related proteins Bax and Bcl-2. (3) SD rats were randomly divided into control group, CKD-VC group and S-adenosylhomocysteine (SAH) intervention group. The calcification of thoracic aorta was evaluated by von Kossa staining, and the protein expressions of Runx2, Bax and Bcl-2 were detected by immunohistochemistry and Western blotting.Results:(1) METTL3 mRNA expression in MHD patients with VC was significantly lower than that in non-VC patients ( P<0.05), and was negatively correlated with coronary artery calcium score ( r=-0.65, P<0.001). (2) The expression of METTL3 in VSMCs stimulated by high phosphorus was decreased and showed a time dependence. Immunofluorescence double label showed that METTL3 and Runx2 were co-expressed in the nucleus. METTL3 was overexpressed in high-phosphorus induced VSMCs, and the expressions of Runx2, collagen I and BMP-2 were significantly decreased, accompanied by the decrease of calcified nodules and Bax/Bcl-2 ratio (all P<0.05). Conversely, METTL3 knockdown aggravated VSMCs calcification by inducing apoptosis. (3) Furthermore, METTL3 inhibitor SAH was administered in vivo, and it was found that inhibition of METTL3 expression significantly increased the calcification of rat thoracic aorta, and the Bax/Bcl-2 ratio and Runx2 expression were up-regulated. Conclusions:Serum METTL3 level is reduced in MHD patients with VC. In vivo and in vitro studies demonstrate that METTL3 inhibits VC in CKD by mediating the apoptosis-related protein Bax/Bcl-2.