OBJECTIVE To investigate the clinical efficacy of pramipexole combined with levodopa-benserazide in the treatment of Parkinson’s disease （PD）. METHODS A total of 108 PD patients treated at the Fifth People’s Hospital of Wuhu City from January 1， 2021， to February 28， 2023， were randomly divided into observation group and control group， with 54 cases in each group. Patients in the control group were administered levodopa-benserazide （initial dose of 62.5 mg per dose）， three times daily； after one month， the dose was increased to 250 mg per dose， four times daily. Patients in the observation group received the same treatment as the control group， with the addition of pramipexole （initial dose of 0.25 mg per dose） orally twice daily on an empty stomach； after 14 days， the dose was increased to 0.25 mg per dose， three times daily. Both groups were treated for 3 months. The short-term efficacy， safety and long-term prognosis of the two groups were compared. RESULTS After treatment， the observation group had significantly lower scores on the Unified Parkinson’s Disease Rating Scale part Ⅲ （UPDRS-Ⅲ）， the Hamilton Depression Scale （HAMD）， the Hamilton Anxiety Scale （HAMA）， and the Parkinson’s Disease Questionnaire-39（ PDQ- 39） compared to the control group； in contrast， the observation group had higher scores on the Montreal Cognitive Assessment （MoCA） scale， the Mini-mental State Examination （MMSE） scale， the Mattis Dementia Rating Scale （DRS）， and the Quality of Life （QOL） scale （P＜0.05）. Both groups showed a significant reduction in UPDRS-Ⅲ and PDQ-39 scores， and a significant increase in DRS scores compared to baseline （P＜0.05）. However， only the observation group showed a significant increase in MoCA scale， MMSE scale， and QOL scores， and a significant decrease in HAMD and HAMA scores compared to baseline （P＜ 0.05）. The total incidence of adverse drug reactions in both groups was not significantly different （P＞0.05）. The 12 months follow-up results showed that the incidence of dementia and mortality rates in the observation group were significantly lower than the control group （P＜0.05）. CONCLUSIONS Pramipexole combined with levodopa-benserazide significantly improves motor function， cognitive function， quality of life and symptoms of depression and anxiety in PD patients， and may reduce the long-term risk of dementia and mortality in these patients.

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 develop GTKO (α-1,3-galactosyltransferase gene-knockout, GTKO)/hCD55 (human CD55) gene-edited xenotransplant donor pigs and verify their function. Methods In this study, CRISPR (clustered regularly interspaced short palindromic repeats)/Cas9 (CRISPR-associated nuclease 9), PiggyBac transposon technology and somatic cell nuclear transfer technology were used to construct GTKO/hCD55 gene-edited Diannan miniature pigs. The phenotype and function of GTKO/hCD55 pigs were analyzed by Sanger sequencing, real-time fluorescence quantitative PCR, flow cytometry, immunofluorescence, bisulfite sequencing, antigen-antibody binding assays, and complement-dependent cytotoxicity assays. Results After transfection of PX458 and PiggyBac gene editing vectors into wild-type fetal pig fibroblasts, 48 single-cell colonies were obtained through puromycin drug screening. Two single-cell colonies were selected for somatic cell nuclear transfer, resulting in two fetal pigs at 33 days of gestation. The GGTA1(α-1,3-galactosyltransferase) genotypes of fetal pig F01 were -17 bp and wild type (WT), while the GGTA1 genotypes of fetal pig F02 were -26 bp/+2 bp and -3 bp. The hCD55 mRNA expression levels of both fetal pigs were significantly higher than those of WT pigs (P＜0.01). The fetal pig F02 was selected as the donor cell source for recloning, 11 surviving piglets were obtained, all identified as GTKO/hCD55 gene-edited pigs. These pigs showed absence of α-Gal antigen expression, but weak or no expression of hCD55 was observed. Methylation analysis of the hCD55 gene's CpG island showed hypermethylation in kidney tissue lacking hCD55 expression, whereas it was not methylated or partially methylated in kidney tissue expressing hCD55. Moreover, codon optimization of the CpG island of the hCD55 gene to reduce CG content could achieve stable expression of the hCD55 gene. In addition, antigen-antibody binding experiment showed that the amount of human IgM binding to GTKO/hCD55 gene-edited pig fibroblasts was significantly lower than that of WT pigs (P＜0.01). Complement-dependent cytotoxicity experiment showed that the survival rate of fibroblasts in GTKO/hCD55 pigs was significantly higher than that in WT pigs (P＜0.01). Conclusion This study demonstrates the successful generation of GTKO/hCD55 gene-edited xenotransplant donor pigs. Methylation-induced gene silencing of the hCD55 gene can be effectively avoided by reducing the CG content of the CpG island through codon optimization. This study provides a reference for the development of xenotransplant donor pigs and guides subsequent research on xenotransplantation.

Abstract To clarify the relationship between sports participation and adolescent suicide risk, as well as its underlying mechanisms for preventing suicidal behaviors, the study employs a literature review approach to analyze, synthesize, and summarize relevant research. The findings reveal that physical exercise can reduce suicide rates among middle school students, with high intensity aerobic exercise performed 5-7 days per week demonstrating the most significant effect. However, daily physical activity and muscle strengthening exercises show no significant correlation with suicide related behaviors. The mechanisms through which physical activity reduces the risk of suicide among adolescents include emotional release leading to self help skills, helping adolescents build social connections, and improving physical and brain function. It is recommended that families and schools help adolescents discover the joy of participating in sports, assist in forming sports teams, and ensure at least 60 minutes of moderate to vigorous physical activity daily to reduce the risk of suicidal behavior among adolescents.

Objective To observe the clinical efficacy of acupuncture at Front Mu point in the treatment of ulcerative colitis.Methods Sixty patients with ulcerative colitis treated at the Traditional Chinese Medicine Hospital in Kunming from August 2022 to June 2023 were collected.Using a random number table method,30 cases were assigned to each of the control group and the combined group.The treatment method involved administering oral mesalazine to the control group for a continuous period of 8 weeks,while the combined group received both oral mesalazine and acupuncture at front Mu points.The clinical efficacy,colonoscopy results score(Baron score),and colonic mucosal healing score(Geboes)before and after treatment were compared.Follow-up was conducted at 3 months to calculate the recurrence rate in the combination and control groups.Results The total effective rate in the combination group was higher than that in the control group(P<0.05),with rates of 93.33%and 67.67%,respectively.After treatment,the disease activity index,Baron score,and Geboes score decreased compared to before treatment(P<0.05),and the combination group had a lower disease activity index,Baron score,and Geboes score than the control group after treatment(P<0.05).Comparing the recurrence rates at 3 months post-treatment,the combination group was lower than the control group.Conclusion Acupuncture at Front Mu Point can significantly improve the clinical symptoms of ulcerative colitis,reduce the recurrence rate compared to patients in the control group,and is safe and reliable without serious adverse reactions.

Objective·To investigate the efficacy and safety of hypertonic dextrose prolotherapy(DPT)in the treatment of postherpetic neuralgia.Methods·Seventy-eight patients with postherpetic neuralgia who visited the Department of Pain of The Affiliated Hospital of Xuzhou Medical University from June 2019 to December 2022 were selected.The patients were randomly assigned to a control group and a research group in a 1∶1 ratio,with 39 patients in each group.The control group was treated with traditional analgesic solution,while the research group was treated with traditional analgesic solution combined with DPT.Visual analog scale(VAS)was used to evaluate the patients'pain level before and after treatment,flow cytometry was used to measure the patients'T-cell subsets,and enzyme-linked immunosorbent assay(ELISA)was used to measure the levels of C-reactive protein(CRP),interleukin-6(IL-6),and IL-10 cytokines.The VAS scores were compared between the two groups of patients before and at 1,2,4,8,and 12 weeks after treatment.CD4+/CD8+,CRP,IL-6,IL-10 levels,and the incidence of adverse reactions before and 2 weeks after treatment were compared between the two groups.Results·There was no statistically significant difference in sex ratio,age,and disease duration between the two groups of patients.The VAS scores of the two groups of patients at 1,2,4,8,and 12 weeks after treatment were significantly lower than those before treatment,and the differences were statistically significant(all P<0.05).The VAS scores of the research group at 1,2,4,8,and 12 weeks after treatment were significantly lower than those of the control group(all P<0.05).There was no statistically significant difference in basal CD4+/CD8+,CRP,IL-6 and IL-10 levels between the two groups of patients.IL-6 and CRP levels in the research group were significantly lower after treatment than those in the control group,and the differences were statistically significant(all P=0.000).CD4+/CD8+ and IL-10 levels were significantly higher in the research group than those in the control group after treatment,and the difference was statistically significant(all P=0.000).No adverse reactions such as local nerve damage,epidural hematoma,infection,pneumothorax or allergy occurred in both groups of patients during the treatment.Conclusion·DPT can significantly reduce the pain of PHN patients,improve patients'T lymphocyte subpopulations and cytokine expression,and can be safely applied to the clinic.

With the advent of an aging society,Alzheimer's disease(AD)has gradually become a major ailment affecting the elderly.AD is a neurodegenerative disorder associated with cognitive impairments.In AD patients,brain network connections are disrupted,and their topological properties are also affected,leading to the disintegration of anatomical and functional connections.Anatomical connections can be tracked and evaluated using structural magnetic imaging(MRI)and diffusion tensor imaging(DTI),while functional connections are detected through functional MRI to assess their connectivity status.This review incorporates the findings of previous scholars and summarizes the current research of AD.It mainly discusses the imaging characteristics of large-scale brain network changes in AD patients,so as to provide researchers with scientific and objective imaging markers for AD prediction and early diagnosis,as well as future research.

BACKGROUND:High tibial osteotomy results in massive blood loss during the perioperative period.Tranexamic acid can effectively reduce perioperative blood loss.However,the method of tranexamic acid application has not been unified. OBJECTIVE:To investigate the effect and safety of different methods of tranexamic acid on perioperative blood loss in the high tibial osteotomy. METHODS:A total of 160 patients who underwent primary unilateral high tibial osteotomy in the Binzhou Medical University Hospital from January 2019 to December 2021,including 69 males and 91 females,were randomly divided into four groups(n=40 per group).Among them,40 patients were given an intravenous infusion of saline containing 2 g tranexamic acid 10 minutes before tourniquet release(venous group);40 patients were given an intravenous infusion of 1 g tranexamic acid and 1 g tranexamic acid was injected through a drainage tube after the closure of the incision(combined group);40 patients were given 2 g tranexamic acid infusion into drainage tube after the closure of the incision(perfusion group);an additional 40 patients were given an intravenous infusion of the same amount of normal saline(blank group).The general information was compared among the four groups of patients.The hemoglobin,hematocrit,intraoperative blood loss,drainage volume,blood transfusion rate,incision complication,and the incidence of deep vein thrombosis were recorded on days 1,3 and 5 after operation in the four groups.The total blood loss and hidden blood loss were calculated. RESULTS AND CONCLUSION:(1)There was no statistically significant difference in general information among the four groups.(2)No significant difference was found in intraoperative blood loss among the four groups.(3)The maximum decreased values of hemoglobin and hematocrit on days 1,3 and 5 after operation,drainage volume,total blood loss and hidden blood loss were all ranked as the combined group

BACKGROUND:With the aging of the global population,the incidence rate of osteoporosis is also increasing.It is very important to further understand its pathogenesis and propose new therapeutic targets.Recent studies have shown that ferroptosis is closely related to the pathogenesis of some bone diseases,such as inflammatory arthritis,osteoporosis and osteoarthritis. OBJECTIVE:To summarize the previous studies on the mechanism of ferroptosis in osteoporosis,so as to provide new therapeutic ideas and potential therapeutic targets for osteoporosis. METHODS:The first author used the computer to search the documents published from 2000 to 2022 in CNKI,WanFang,VIP,PubMed and Web of Science with the key words of"ferroptosis,osteoporosis,osteoblasts,osteoclasts,iron chelators,reactive oxygen species,nuclear factor erythroid 2-related factor 2,heme oxygenase-1,glutathione peroxidase 4,review"in Chinese and English.A total of 70 articles were finally included according to the inclusion criteria. RESULTS AND CONCLUSION:Ferroptosis is significantly different from necrosis,apoptosis and autophagy.In terms of cell morphology and function,it does not have the morphological characteristics of typical necrosis,nor does it have the characteristics of traditional apoptosis,such as cell contraction,chromatin condensation,the formation of apoptotic bodies and the disintegration of cytoskeleton.Contrary to autophagy,ferroptosis does not form a classical closed bilayer membrane structure(autophagic vacuole).Morphologically,ferroptosis is mainly manifested by obvious contraction of mitochondria,increased membrane density,and reduction or disappearance of mitochondrial cristae,which are different from other cell death modes.Iron overload can destroy bone homeostasis by significantly inhibiting osteogenic differentiation and stimulating osteoclast formation,leading to osteoporosis.Iron overload interferes with the differentiation of stem cells to osteoblasts,leading to a weakened osteoblast function and further imbalance of bone metabolism in the body,which eventually leads to osteoporosis.Stimulated by iron overload,osteoclast bone resorption is enhanced and bone loss exceeds new bone formation.Iron chelators have been proved to have osteoprotective effects by inhibiting osteoclast activity and stimulating osteogenic differentiation of osteoblasts.Its potential mechanism is related to inhibiting osteoclast differentiation and promoting osteoblast differentiation.Antioxidants can prevent reactive oxygen species production and inhibit bone absorption,thus improving bone metabolism and effectively preventing osteoporosis.

Pharmacological perturbation studies based on protein-level signatures are fundamental for drug dis-covery.In the present study,we used a mass spectrometry(MS)-based proteomic platform to profile the whole proteome of the breast cancer MCF7 cell line under stress induced by 78 bioactive compounds.The integrated analysis of perturbed signal abundance revealed the connectivity between phenotypic behaviors and molecular features in cancer cells.Our data showed functional relevance in exploring the novel pharmacological activity of phenolic xanthohumol,as well as the noncanonical targets of clinically approved tamoxifen,lovastatin,and their derivatives.Furthermore,the rational design of synergistic inhibition using a combination of histone methyltransferase and topoisomerase was identified based on their complementary drug fingerprints.This study provides rich resources for the proteomic landscape of drug responses for precision therapeutic medicine.