Objective To explore the construction of α-1,3-galactosyltransferase (GGTA1) gene-knockout (GTKO) Diannan miniature pigs and the kidney xenotransplantation from pigs to rhesus macaques, and to assess the effectiveness of GTKO pigs. Methods The GTKO Diannan miniature pigs were constructed using the CRISPR/Cas9 gene-editing system and somatic cell cloning technology. The phenotype of GTKO pigs was verified through polymerase chain reaction, Sanger sequencing and immunofluorescence staining. Flow cytometry was used to detect antigen-antibody (IgM) binding and complement-dependent cytotoxicity. Kidney xenotransplantation was performed from GTKO pigs to rhesus macaques. The humoral immunity, cellular immunity, coagulation and physiological indicators of the recipient monkeys were monitored. The function and pathological changes of the transplanted kidneys were analyzed using ultrasonography, hematoxylin-eosin staining, immunohistochemical staining and immunofluorescence staining. Results Single-guide RNA (sgRNA) targeting exon 4 of the GGTA1 gene in Diannan miniature pigs was designed. The pGL3-GGTA1-sgRNA1-GFP vector was transfected into fetal fibroblasts of Diannan miniature pigs. After puromycin selection, two cell clones, C59# and C89#, were identified as GGTA1 gene-knockout clones. These clones were expanded to form cell lines, which were used as donor cells for somatic cell nuclear transfer. The reconstructed embryos were transferred into the oviducts of trihybrid surrogate sows, resulting in 13 fetal pigs. Among them, fetuses F04 and F11 exhibited biallelic mutations in the GGTA1 gene, and F04 had a normal karyotype. Using this GTKO fetal pig for recloning and transferring the reconstructed embryos into the oviducts of trihybrid surrogate sows, seven surviving piglets were obtained, all of which did not express α-Gal epitope. The binding of IgM from the serum of rhesus monkey 20# to GTKO pig PBMC was reduced, and the survival rate of GTKO pig PBMC in the complement-dependent cytotoxicity assay was higher than that of wild-type pig. GTKO pig kidneys were harvested and perfused until completely white. After the left kidney of the recipient monkey was removed, the pig kidney was heterotopically transplanted. Following vascular anastomosis and blood flow restoration, the pig kidney rapidly turned pink without hyperacute rejection (HAR). Urine appeared in the ureter 6 minutes later, indicating successful kidney transplantation. The right kidney of the recipient was then removed. Seven days after transplantation, the transplanted kidney had good blood flow, the recipient monkey's serum creatinine level was stable, and serum potassium and cystatin C levels were effectively controlled, although they increased 10 days after transplantation. Seven days after transplantation, the levels of white blood cells, lymphocytes, monocytes and eosinophils in the recipient monkey increased, while platelet count and fibrinogen levels decreased. The activated partial thromboplastin time, thrombin time and prothrombin time remained relatively stable but later showed an upward trend. The recipient monkey survived for 10 days. At autopsy, the transplanted kidney was found to be congested, swollen and necrotic, with a small amount of IgG deposition in the renal tissue, and a large amount of IgM, complement C3c and C4d deposition, as well as CD68⁺ macrophage infiltration. Conclusions The kidneys of GTKO Diannan miniature pigs may maintain normal renal function for a certain period in rhesus macaques and effectively overcome HAR, confirming the effectiveness of GTKO pigs for xenotransplantation.

Objective To observe the effects of tuina on P2Y12/RhoA/ROCK2 pathway and c-Fos protein expression of microglia in spinal dorsal horn of sciatic nerve chronic compressive injury(CCI)rats;To explore the mechanism of tuina in treating lumbar disc herniation.Methods A CCI model of right sciatic nerve was used to simulate neuropathic pain in lumbar disc herniation.24 male SD rats were randomly divided into blank group,model group and tuina group,with 8 rats in each group.After 4 days of modeling,massage intervention was used to in the tuina group for 14 days.The paw withdrawal threshold(PWT)and paw withdrawal latency(PWL)of rats before and on the 4th,10th,and 17th day after modeling were observed,immunofluorescence staining was used to detect the expression of Iba1 and P2Y12 protein in right spinal dorsal horn,Western blot was used to detect the expressions of RhoA and ROCK2 protein in right spinal dorsal horn,immunohistochemistry staining was used to detect the number of c-Fos positive cells in right spinal dorsal horn.Results Compared with the blank group,the PWT and PWL of the model group were significantly reduced on the 4th,10th,and 17th day after modeling(P<0.001),while the expressions of Iba1,P2Y12,RhoA and ROCK2 protein in right spinal dorsal horn significantly increased(P<0.001,P<0.05),the number of c-Fos positive cells significantly increased(P<0.001).Compared with the model group,the PWT and PWL of the tuina group significantly increased on the 10th and 17th day after modeling(P<0.05,P<0.01,P<0.001),and the expressions of Iba1,P2Y12,RhoA and ROCK2 protein in right spinal dorsal horn significantly decreased(P<0.001,P<0.01,P<0.05),and the number of c-Fos positive cells significantly reduced(P<0.001).Conclusion Tuina may inhibit the activation of microglia by regulating P2Y12/RhoA/ROCK2 pathway and c-Fos protein expression in spinal dorsal horn,reduce neuronal excitability,and exert analgesic effects on lumbar disc herniation.

ObjectiveTo explore the possible mechanism of Tuina at Weizhong （BL 40） for relieving sciatica from the perspective of hippocampal synaptic plasticity. MethodsSixty SD rats were randomly divided into sham operation group， model group， Tuina group， MK-801 group， MK-801 plus Tuina group， 12 rats in each group. After lateral ventricular cannulation， rats model with chronic compression injury of the right sciatic nerve were prepared in all groups except the sham operation group. On day 4 after modelling， rats in the Tuina group start Tuina at Weizhong （BL 40） for 10 mins once a day for a total of 14 days； rats in the MK-801 group started injecting with 0.25 μg/μl of the N-methyl-D-aspartate receptor 2B （NR2B） blocker， dizocycline （MK-801）， 0.5 μl of which was administered daily in the lateral ventricle for 14 days. Rats in the MK-801 plus Tuina group underwent Tuina after 30 mins when completing MK-801 injection in the lateral ventricle， in the same way as above； rats in the model group and the sham operation group did not undergo any intervention. Spontaneous pain behaviour scores and paw withdraw thresholds （PWTs） were examined on day 1 （base value） before modelling and on day 4， 10， 14 and 18 after modelling； and on day 19， the brain tissues of the rats in each group were sampled and the number and morphology of the Nysted-positive cells in the hippocampal CA3 region were observed using Nysted staining； and the number of synapses， the thickness of postsynaptic dense material， the length of active band and the curvature of synaptic interface in hippocampal CA3 region were observed by transmission electron microscopy； and the expression of synapse-associated proteins NR2B and postsynaptic density protein-95 （PSD95） in hippocampal CA3 was detected by immunofluorescence staining and immunoblotting. ResultsCompared with the same time in the sham operation group， spontaneous pain scores significantly increased and PWTs decreased on day 4， 10， 14， and 18 after modelling in the model group （P＜0.05）； compared with the model group， spontaneous pain scores in Tuina group of rats significantly decreased on day 10， 14， and 18 after modelling， and PWTs significantly increased on day 14 and 18 after modelling （P＜0.05）. Compared with Tuina group， spontaneous pain scores increased on day 10， 14， and 18 of modelling， and PWTs decreased at days 14 and 18 of modelling in the MK-801 plus Tuina group had higher spontaneous pain scores on days 10， 14， and 18 after modelling and lower PWTs on days 14 and 18 after modelling （P＜0.05）. Compared with the sham operation group， the neuronal arrangement in the hippocampal CA3 region of the rats in the model group was disordered， with decreased number of Nysted-positive cells and synapses， reduced thickness of postsynaptic densities， length of active bands， and curvature of synaptic interfaces， wider synaptic gaps， and decreased immunofluorescent positive expression of NR2B and PSD95 as well as the expression of immunoblotting proteins in hippocampal CA3 region （P＜0.05）. Compared with the model group， more dense arranged nerve cells， the number of Nysted-positive cells， the number of synapses， the thickness of postsynaptic dense material， the length of active bands increased， the synaptic gap became significantly narrower， and the positive expression of immunofluorescence and immunoblotting protein expression of NR2B， PSD95 increased in the rat hippocampal CA3 region of Tuina group （P＜0.05）. Compared with Tuina group， the neuronal morphology of the hippocampal CA3 region in MK-801 plus Tuina group was severely damaged， and the number of Nystrom's-positive cells， the number of synapses， the thickness of post-synaptic densities， the length of active bands， and the curvature of synaptic interfaces reduced， the synaptic gaps became wider， and the immunofluorescent positive expression of NR2B， PSD95， and the expression of immunostained proteins decreased （P＜0.05）. ConclusionTuina at "Weizhong" （BL 40） showed significant analgesic effect， and one of the possible mechanisms concluded as significantly increasing the levels of NR2B and PSD95 protein expression in hippocampal CA3 region and thus modulating the synaptic plasticity of the hippocampus.

Chronic heart failure is the terminal stage of progressive aggravation of various cardio-vascular diseases,and as an independent risk factor for heart failure,senility can increase the preva-lence rate.Ventricular remodeling is the main pathological and physiological basis of chronic heart failure,and in addition to changes in the activities of inflammatory reactions and related cytokines,factors such as metabolic disorders of myocardium,regulation of myocardial extracellular vesicles,and imbalance of intestinal microbiota also play important roles.This paper provided an overview of the research progress on ventricular remodeling mechanism in elderly patients with heart failure.

Objective: To investigate the analgesic mechanism of Tuina (Chinese therapeutic massage) by observing the effect of the N-methyl-D-aspartate receptor subunit 2B (NR2B)/postsynaptic density-95 (PSD-95) pathway on the dendritic structure of spinal cord dorsal horn in rats with lumbar disc herniation. Methods: Fifty Sprague-Dawley rats were randomly divided into a blank group, a model group, a Tuina group, a blocker agent group, and a blocker agent + Tuina group. The sciatic nerve chronic constriction injury (CCI) model was prepared by the sciatic nerve ligation method. From the 4th day after modeling, rats in the Tuina group and the blocker agent + Tuina group were subject to daily Tuina intervention, and those in the blocker agent group and the blocker agent + Tuina group were daily intrathecally injected with NR2B blocker agent (MK-801). The spontaneous pain score was used to observe the pain behavior of all rats. The expression levels of NR2B and downstream PSD-95 were measured by immunohistochemistry, and the dendritic structure changes were observed by Golgi staining for rat spinal cord dorsal horn after 14 d of continuous intervention. Results: Compared with the blank group, the degree of rat spontaneous pain after CCI was elevated in both the model and the Tuina groups (P<0.01) and was reduced in the Tuina group after the Tuina intervention compared with the model group (P<0.05). Compared with the model group, the rat spontaneous pain level after blocking NR2B was reduced in both the blocker agent group and the blocker agent + Tuina group (P<0.05). The NR2B and PSD-95 protein levels were significantly higher in the model group compared with the blank group (P<0.01); the total number of dendritic branches was increased (P<0.01), and the total dendritic length became longer (P<0.01) in the spinal cord dorsal horn. The rat NR2B and PSD-95 protein levels were significantly decreased in the Tuina group compared with the model group (P<0.01); the total dendritic branch number was reduced (P<0.01) and the total length was shortened (P<0.01) in the spinal cord dorsal horn. After blocking NR2B, the expression levels of NR2B and downstream PSD-95 protein were significantly lower in both the blocker agent group and the blocker agent + Tuina group compared to the model group (P<0.01). The total branch number was significantly reduced (P<0.01), and the total length was significantly shortened (P<0.01) of the dendrites in the spinal cord dorsal horn. Conclusion: Tuina may exert an analgesic effect by remodeling the dendritic structure in the spinal cord dorsal horn in rats with lumbar disc herniation, and its mechanism may be related to the inhibition of NR2B/PSD-95 signaling pathway.

Objective:To explore the efficacy of ixazomib combination treatment therapies for multiple myeloma (MM), and the influencing factors of prognosis.Methods:The clinical data of 80 MM patients admitted to Yancheng Third People's Hospital from January 2020 to January 2022 were retrospectively analyzed. All patients received 3 courses of ixazomib combination treatment therapies (28 d was 1 course). All combination treatment therapies included ID group (ixazomib + dexamethasone, 11 cases) and ID + immunomodulator group (ixazomib + dexamethasone + lenalidomide/thalidomide, 50 cases), ID + other chemotherapy drugs group (ixazzomib + dexamethasone + doxorubicin liposome/cyclophosphamide/bendamustine, 19 cases). The clinical efficacy of patients in different treatment regimens was compared, and the prognosis was followed up and recorded. The clinical characteristics between the survival and the dead patients were compared. Cox proportional risk model was used to make multivariate analysis of the overall survival of MM patients receiving ixazomib combination therapies.Results:The treatment was effective in 9 cases (81.82%) of the ID group, 32 cases (64.00%) of the ID + immunomodulator group, and 9 cases (47.37%) of the ID + other chemotherapy drugs group. There was no statistically significant difference in the effectiveness rate of 3 ixazomib combination regimens ( χ2 = 0.62, P = 0.432). All patients were followed up for 5 to 20 months, with an average follow-up time of (15±4) months. There were statistically significant differences in immunoglobulin type, Durie-Salmon stage, early treatment line and therapeutic efficacy between the survival group (49 cases) and the death group (31 cases) (all P < 0.05). Multivariate Cox regression analysis showed that the clinical effectiveness (effectiveness vs. ineffectiveness: OR = 0.242, 95% CI 0.103-0.567, P = 0.001) and the previous first-line treatment (the first-line vs. the other lines: OR = 0.577, 95% CI 0.452-0.736, P < 0.001) were independent protective factors for the overall survival of MM. Conclusions:The 3 ixazomib combination therapies have a certain efficacy in the treatment of MM; ID regimen has the best clinical efficacy and survival. The clinical effectiveness and the previous first-line treatment are independent protective factors for the survival of MM.

OBJECTIVE To review the application and development of automatic intelligent drug cabinet in hospitals at home and abroad. METHODS The relevant research articles published from 2010 to 2022 were retrieved from CNKI, Wanfang, VIP, PubMed and other databases. RESULTS The application of automatic intelligent drug cabinet could shorten the execution time of inpatient medical orders, improve the work efficiency of pharmacists and doctors and nurses, improve the quality of drug management in wards and promote the transformation of hospital pharmaceutical care. CONCLUSION The construction of decentralized dispensing mode with automated intelligent drug cabinets as the core has important theoretical guiding significance and broad application prospects for the automation, informatization, intelligence and whole process management of drugs in medical institutions, and also helps to improve the level of medical pharmacy service management.

Chronic heart failure is the terminal stage of progressive aggravation of various cardio-vascular diseases,and as an independent risk factor for heart failure,senility can increase the preva-lence rate.Ventricular remodeling is the main pathological and physiological basis of chronic heart failure,and in addition to changes in the activities of inflammatory reactions and related cytokines,factors such as metabolic disorders of myocardium,regulation of myocardial extracellular vesicles,and imbalance of intestinal microbiota also play important roles.This paper provided an overview of the research progress on ventricular remodeling mechanism in elderly patients with heart failure.

Objective:To evaluate the therapeutic effect of radiofrequency ablation in the treatment of severe hemangioma with thrombocytopenia and coagulation abnormality (Kasabach-Merritt phenomenon, KMP).Methods:From April 2018 to April 2021, 11 cases of KMP in our department were retrospectively analyzed. The patients underwent MRI, color ultrasound and hematology examination. Radiofrequency ablation was performed for all patients under general anesthesia. The comprehensive, multi-level and multi-point ablation was accurately implemented under the whole-process intraoperative monitoring of color ultrasound. Postoperative ice pad was applied for the treatment area, and dressing change was carried out every day.Results:All 11 cases of KMP were treated with radiofrequency ablation, including 5 males and 6 females, aged 1 d to 2 months, with a median age of 15 d. Intraoperative color Doppler ultrasound monitoring showed that the tumor was completely ablated in the single operation. All patients were followed up for 0.5 to 2 years. The tumor was cured without recurrence. The local function was unaffected. The local scar formation was mild and the appearance was satisfactory.Conclusions:Radiofrequency ablation has great clinical value in the treatment of KMP. It makes up the shortcomings and avoids the disadvantages of traditional surgery and other conservative treatment methods. Good clinical results can be achieved by radiofrequence ablation with satifactory function and appearance.

Objective:To investigate the outcomes of therapeutic radiofrequency ablation (RFA) in unilateral intra-articular venous malformations (IAVMs) of the knee.Methods:The clinical data of 15 patients with intravascular venous malformation in the Third Affiliated Hospital of Zhengzhou University from February 2018 to May 2019 were retrospectively analyzed. There were 7 males and 8 females with an average age of 16 years (range, 5—25 years). All patients had unilateral knee involvement, ranging from localized lesions to diffuse extensive lesions. The radiofrequency ablation electrode needle with a diameter of 0.5 mm was selected and the power was adjusted to 25 W. Under the guidance of color ultrasound monitoring, the whole operation was carried out. The RFA electrode needle was inserted into the deepest site of the lesion and extracted by 1-mm graded movements until a point 1 mm from the surface of the lesion. Each ablation point was ablated for 10 to 30 seconds and multi-point ablation was done until the lesions were completely eliminated. Patients were followed up for 6 months to 1 year. Treatment outcomes were evaluated by functional activity, including extension, flexion, abduction, and adduction at the knee joint, and MRI findings, including intensity, shape, boundary, and degree of aggressiveness in the intra- and extraarticular cavities.Results:Included 15 patients; 12 had VMs in the knee joint cavity, and 3 had VMs involving both the intraarticular and extraarticular cavities (1 from the joint cavity to the calf, 1 from the joint cavity to the lower femur, and 1 from the intraarticular to the extraarticular cavity). All subjects underwent RFA, and the lesions were completely ablated and eradicated in a single operation. No recurrence was reported in the 6 months to 1 year follow-up. The knee joint remained symmetrical with the contralateral knee joint, without scarring, after the RFA.Conclusions:Radiofrequency ablation for the treatment of intra-articular venous malformation of the knee joint is a valuable minimally invasive surgical method with complete ablation, no complications, good prognosis and aesthetic appearance.