Aim To investigate the role of metabolites of eicosapentaenoic acid (EPA) in promoting the transdifferentiation of pancreatic α cells to β cells. Methods Male C57BL/6J mice were injected intraperitoneally with 60 mg/kg streptozocin (STZ) for five consecutive days to establish a type 1 diabetes (T1DM) mouse model. After two weeks, they were randomly divided into model groups and 97% EPA diet intervention group, 75% fish oil (50% EPA +25% DHA) diet intervention group, and random blood glucose was detected every week; after the model expired, the regeneration of pancreatic β cells in mouse pancreas was observed by immunofluorescence staining. The islets of mice (obtained by crossing GCG

Aim To explore the possible mechanism of "component-target-pathway" of Radix Hedysari against target organ damage caused by radiotherapy and chemotherapy, and to verify the " dose-effect" relationship of the main active components. Methods TCMSP, Uniprot, Swiss Target Prediction, GeneCards, Cytoscape, Omicshare and other platforms were used for network pharmacology analysis. Autodock, Pymol and Ligplot were used for molecular docking. The water extract of Radix Hedysari was used for animal experiment verification. The contents of eight main components were determined by HPLC. Results Four active components, eight key targets and four key pathways of Radix Hedysari were identified to resist the damage of target organs caused by radiotherapy and chemotherapy. Molecular docking showed that formononetin and quercetin had good binding activity with HSP90AA1, naringenin and MAPK3, and ursolic acid and TP53. Animal experiments showed that gastrointestinal factors MTL and VIP increased significantly, liver and kidney factors Cr, BUN, AST and ALT decreased significantly, inflammatory factor IL-10 increased significantly and TNF-a decreased significantly. The content of ononm was the highest (2 . 884 8 µg • g "

Objective Recombinase-aided polymerase chain reaction(RAP)is a sensitive,single-tube,two-stage nucleic acid amplification method.This study aimed to develop an assay that can be used for the early diagnosis of three types of bacteremia caused by Staphylococcus aureus(SA),Pseudomonas aeruginosa(PA),and Acinetobacter baumannii(AB)in the bloodstream based on recombinant human mannan-binding lectin protein(M1 protein)-conjugated magnetic bead(M1 bead)enrichment of pathogens combined with RAP. Methods Recombinant plasmids were used to evaluate the assay sensitivity.Common blood influenza bacteria were used for the specific detection.Simulated and clinical plasma samples were enriched with M1 beads and then subjected to multiple recombinase-aided PCR(M-RAP)and quantitative PCR(qPCR)assays.Kappa analysis was used to evaluate the consistency between the two assays. Results The M-RAP method had sensitivity rates of 1,10,and 1 copies/μL for the detection of SA,PA,and AB plasmids,respectively,without cross-reaction to other bacterial species.The M-RAP assay obtained results for<10 CFU/mL pathogens in the blood within 4 h,with higher sensitivity than qPCR.M-RAP and qPCR for SA,PA,and AB yielded Kappa values of 0.839,0.815,and 0.856,respectively(P<0.05). Conclusion An M-RAP assay for SA,PA,and AB in blood samples utilizing M1 bead enrichment has been developed and can be potentially used for the early detection of bacteremia.

Objective To explore the biomarkers and potential mechanisms of chronic restraint stress-induced myocardial injury in hyperlipidemia ApoE-/-mice.Methods The hyperlipidemia combined with the chronic stress model was established by restraining the ApoE-/-mice.Proteomics and bioinformatics techniques were used to describe the characteristic molecular changes and related regulatory mechanisms of chronic stress-induced myocardial injury in hyperlipidemia mice and to explore potential diagnostic biomarkers.Results Proteomic analysis showed that there were 43 significantly up-regulated and 58 sig-nificantly down-regulated differentially expressed proteins in hyperlipidemia combined with the restraint stress group compared with the hyperlipidemia group.Among them,GBP2,TAOK3,TFR1 and UCP1 were biomarkers with great diagnostic potential.KEGG pathway enrichment analysis indicated that fer-roptosis was a significant pathway that accelerated the myocardial injury in hyperlipidemia combined with restraint stress-induced model.The mmu_circ_0001567/miR-7a/Tfr-1 and mmu_circ_0001042/miR-7a/Tfr-1 might be important circRNA-miRNA-mRNA regulatory networks related to ferroptosis in this model.Conclusion Chronic restraint stress may aggravate myocardial injury in hyperlipidemia mice via ferrop-tosis.Four potential biomarkers are selected for myocardial injury diagnosis,providing a new direction for sudden cardiac death(SCD)caused by hyperlipidemia combined with the restraint stress.

Objective To investigate the effect of Polyetheretherketone(PEEK)rod semi-rigid pedicle screw fixation sys-tem in lumbar spine non-fusion surgery.Methods A total of 74 patients with tow-level lumbar degenerative diseases who un-derwent surgery from March 2017 to December 2019 were divided into PEEK rod group and titanium rod group.In the PEEK rod group,there were 34 patients,including 13 males and 21 females,aged from 51 to 79 years old with an average of(62.4±6.8)years old;There were 1 patient of L1-L3 segments,7 patients of L2-L4 segments,20 patients of L3-L5 segments and 6 pa-tients of L4-S1 segments.In the titanium rod group,there were 40 patients,including 17 males and 23 females,aged from 52 to 81 years old with an average of(65.2±7.3)years old;There were 3 patient of L1-L3 segments,11 patients of L2-L4 segments,19 patients of L3-L5 segments and 7 patients of L4-S1 segments.The general conditions of operation,such as operation time,intraoperative blood loss,postoperative drainage was recorded.The visual analogue scale(VAS)for low back pain and Os-westry disability index(ODI)were compared in preoperatively and postoperatively(3 months,12 months and last follow-up)between two groups.The change of range of motion(ROM)was observed by flexion and extension x-ray of lumbar Results All patients successfully completed the operation.The follow-up time ranged from 22 to 34 months with an average of(26.8±5.6)months.The operative time(142.2±44.7)min and intraoperative blood loss(166.5±67.4)ml in PEEK group were lower than those in titanium group[(160.7±57.3)min、(212.8±85.4)ml](P＜0.05).There was no significant differences in postoperative drainage between the two groups(P＞0.05).At the final follow-up visit,in PEEK group and titanium group VAS of low back pain[(0.8±0.4)points vs(1.0±0.5)points],VAS for leg pain[(0.7±0.4)points vs(0.8±0.5)points]and ODI[(9.8±1.6)％vs(12.1±1.5)％]were compared with preoperative[(5.8±1.1)points vs(6.0±1.1)points],[(7.2±1.7)points vs(7.0±1.6)points],[(68.5±8.9)％vs(66.3±8.2)％]were significantly different(P＜0.05).There was no significant difference in VAS scores between the two groups at each postoperative time point(P＞0.05).At 3 months after surgery,there was no difference in ODI between the two groups(P＞0.05).There were significant differences in ODI between PEEK group and titanium rod group at 12 months[(15.5±2.1)％vs(18.4±2.4)％]and at the last follow-up[(9.8±1.6)％vs(12.1±1.5)％](P＜0.05).The total range of motion(ROM)of lumbar decreased in both groups after surgery.At 12 months after surgery and the last follow-up,the PEEK group compared with the titanium rod group,the total range of motion of lumbar was statistically significant(P＜0.05).The range of motion(ROM)of the fixed segments decreased in both groups after surgery.The ROM of the fixed segments in PEEK group decreased from(9.5±4.6)° to(4.1±1.9)° at the last follow-up(P＜0.05),which in the titanium rod group was de-creased from(9.8±4.3)°to(0.9±0.5)° at the last follow-up(P＜0.05).The range of motion(ROM)of upper adjacent segment increased in both groups,there was statistical significance in the ROM of upper adjacent segment between the two groups at 12 months after surgery and the last follow-up,(P＜0.05).There was no screw loosening and broken rods in both groups during the follow-up period.Conclusion The PEEK rod semi-rigid pedicle screw internal fixation system used in lumbar non-fusion surgery can retain part of the mobility of the fixed segment,showing comparable short-term clinical efficacy to titanium rod fu-sion.PEEK rod semi-rigid pedicle screw internal fixation system is a feasible choice for the treatment of lumbar spine degener-ative diseases,and its long-term efficacy needs further follow-up observation.

Aim To observe the effects of Compound Dihuang Granules on α-syn,VAPB and PTPIP51 in the substantia nigra of Parkinson's disease(PD)rats with Yin deficiency and wind syndrome,and to explore the possible mechanisms of their actions.Methods The 6-hydroxydopamine-induced PD model of rats was constructed.The model rats were randomly divided into the model group,madopar group,CLD group,CMD group and CHD group,while the NC group did not re-ceive any treatment and the SO group was injected with ascorbic acid,with 13 rats in each group.The neurobe-havioral changes of the rats were observed,and the ex-pressions of α-syn,VAPB and PTPIP51 in the sub-stantia nigra were detected by Western blot,RT-PCR and Immunohistochemistry;the histopathological and morphological changes of the substantia nigra tissue were observed by HE staining,the changes of mito-chondrial ultrastructure in the substantia nigra cells were observed by transmission electron microscopy,and the changes of ATP content in substantia nigra tis-sue in each group were detected by ELISA.Results Compared with the NC and SO groups,rats in the model group showed that the number of rotational cir-cles and pole-climbing time increased,the expression of α-syn increased,the expression of VAPB and PTPIP 5 1 decreased,the number of neuronal cells decreased,the neuronal cells became crumpling,and mitochondrial swelling,disappearance of the mitochon-drial cristae,a larger distance between the ER-mito-chondrial contacts were observed;the ATP content de-creased.Compared with the model group,rats in Mado-par group and CLD group,CMD group and CHD group showed that the number of rotational circles and pole-climbing time decreased,the expression of α-syn de-creased,the expression of VAPB and PTPIP51 in-creased,the degree of neuronal damage was reduced,the morphology of mitochondria was improved and the content of ATP increased,showing the change of the difference in quantitative efficacy;the relative efficacy of Madopar group and CHD group was better,and there was no statistically significant difference.Con-clusions Compound Dihuang granules attenuate the behavioral symptoms in PD rats and may play a thera-peutic role in PD by down-regulating the expression ofα-syn,up-regulating the expression of PTPIP51 and VAPB and improving mitochondrial function.

Aim To investigate the preventive and therapeutic effects of the mfat-1 gene therapy on exper-imental autoimmune encephalomyelitis in mice.Meth-ods mfat-1 gene therapy was used to render the host endogenous capability of producing ω-3 PUFAs,con-comitantly reduce the levels of ω-6 PUFAs,and change the proportion of ω-3/ω-6 PUFAs.Then,the levels of PUFAs in blood were analyzed by gas chromatography.The neurological deficits in mice were evaluated by neurological dysfunction score.HE staining and LFB staining of mouse spinal cord slices were used to ob-serve central nervous system inflammation infiltration and demyelinating lesions.Flow cytometry microsphere microarray technology was used to detect the content of cytokines in serum.Results The mfat-1 gene therapy could significantly raise the proportion of ω-3/ω-6 PU-FAs(P＜0.05),markedly delay the incubation period and peak period and reduce neurological dysfunction scores(P＜0.05),and improve inflammation and de-myelination of spinal cords(P＜0.05).It could also greatly increase the levels of IL-2,IFN-γ,IL-4 and IL-17 in serum(P＜0.05).Conclusion The pro-portion of ω-3/ω-6 PUFAs in blood circulation en-hanced by mfat-1 gene therapy can effectively prevent and treat experimental autoimmune encephalomyelitis in mice.

This study performed to determine whether OX40L overexpressed by recombinant rabies virus(LBNSE-OX40L)can enhance the innate immune response through activation of dendritic cells and thus activate early antibody production.Bone marrow dendritic cells(BMDCs)were extracted from the femur of Balb/c mice and cultured for 6 days,and the cultured BMDCs were infected with the parental virus LBNSE and the recombinant virus LBNSE-OX40L with the multiplicity of infections(MOI)=1.The effect of each virus on the maturation of BMDCs was analyzed by flow cytometry;ELISA was used to detect the expression of innate immunity-related cytokines such as interferon-α(IFN-α)and interleukin-12p40(IL-12p40)in the supernatants of the infected BMDCs.For in vivo study,Balb/c female mice were injected intramuscularly with 106 FFU of parental virus LBNSE and recombinant virus LBNSE-OX40L in both hind limbs,and the inguinal lymph nodes of mice were collected on day 6 after immunization,and the proportion of mature DCs was detected by flow cytometry.The serum was collected on day 6 after immunization,and the content of virus-neutralizing antibody(VNA)was detected by antiviral neutralizing antibody titration.Mouse serum was collected on day 6 after immunization,and virus neutralizing antibody content was measured by titration of antiviral neutralizing antibody,while IgG antibody in mouse serum was detected by ELISA.IgM antibody subclasses were detected by ELISA on days 2,4,and 6 after immunization.Compared with the parental virus LBNSE,the recombinant virus LBNSE-OX40L was able to activate more BMDCs in vitro and produce significantly higher levels of IFN-α and IL-12p40.Furthermore,the recombinant virus LBNSE-OX40L stimulated the maturation and differentiation of the DCs in vivo,which led to the rapid production of high levels of VNA and RABV-specific IgG and IgM antibodies.Taken together,LBNSE-OX40L activates dendritic cells to promote the body's innate immune response,and in turn enhances early antibody production,thus can be an early effective rabies vaccine candidate.

Objective:To compare the effects of transurethral resection of the prostate(TURP)and transurethral columnar bal-loon dilatation of the prostate(TUCBDP)in the treatment of BPH.Methods:This study included 218 BPH patients treated in Qin-huangdao Workers'Hospital from July 2021 to November 2022,109 by TURP and the other 109 by TUCBDP.We followed up the patients for 12 months,observed their postoperative recovery,complications,serum pain,inflammatory index,cytokine level,urodynamic index,symptom improvement and quality of life(QOL)and compared the data obtained between the two groups of patients.Results:At 12 months after surgery,the total effectiveness rate was significantly higher in the TUCBDP than in the TURP group(93.58%vs 84.40%,P<0.05),and the postoperative recovery was better in the former than in the latter(P<0.05).Compared with the baseline,the lev-els of serum prostaglandin E2(PGE2),substance P,tumor necrosis factor-alpha(TNF-α)and high sensitive C-reactive protein(hs-CRP)were remarkably increased in both of the groups on the first day after surgery(P<0.05),more significantly in the TURP than in the TUCBDP group(P<0.05),while the levels of serum PSA and E2 decreased and the T level elevated in all the patients at 3 months postoperatively(P<0.05),more significantly in the TUCBDP than in the TURP group(P<0.05).Before and at 3 and 12 months af-ter operation,the postvoid residual urine volume(PVR)and NIH-CPSI,IPSS and QOL scores showed a decreasing trend,while the maximum urinary flow rate(Qmax),maximum cystometric capacity(MCC)and maximum urethral closure pressure(MUCP)exhibited an increasing trend in both of the two groups,even more significantly in the TUCBDP than in the TURP group(P<0.05).Conclu-sion:TUCBDP is advantageous over TURP in promoting postoperative recovery,improving QOL,reducing postoperative pain,inflamma-tion and complications,regulating the levels of serum cytokines,and improving urodynamics and clinical symptoms in BPH patients.However,with the extension of postoperative time,the two strategies are basically comparable in improving the urodynamics,symptoms and QOL of the patients.

AIM To investigate the effects of Compound Dihuang Granules on biological function of dopaminergic neurons in rats with Parkinson's disease(PD)of Pattern of Yin-Deficiency with Stirring Wind.METHODS The PD animal model of Pattern of Yin-Deficiency with Stirring Wind established by intracerebral injection of 6-hydroxydopamine(6-OHDA)were randomly divided into the model group,the Midoba group(150 mg/kg)and the low,medium and high dose Compound Dihuang Granules groups(1.75,3.5 and 7 g/kg)for corresponding drug intervention,in contrast to the normal group,the sham operation group and the model group underwent 28-day normal saline administration.The rats had their general condition and neuroethology observed;their pathological changes of substantia nigra observed by HE staining;their mitochondrial structure of dopaminergic neurons in the damaged substantia nigra observed by transmission electron microscopy;and their expressions of DJ-1,IP3R,GRP75 and VDAC1 detected by immunohistochemistry,Western blot and RT-qPCR.RESULTS Compared with the normal group and sham operation group,the model group displayed increased rotational behavirors(P＜0.01),decreased swimming time score(P＜0.01),decreased hanging time(P＜0.01),decreased number of neurons in substantia nigra but more neurons with morphological damage,mitochondrial swelling and degeneration,mitochondrial crista disappearance,and decreased expressions of DJ-1,IP3R,GRP75 and VDA C1 mRNA and protein in the injured side(P＜0.01).Compared with the model group,the Midoba group and the Compound Dihuang Granules groups demonstrated less rotational behavirors(P＜0.01),higher swimming time score(P＜0.01),longer hanging time(P＜0.01),less damage to mitochondrial morphology and structure,and higher expressions of DJ-1,IP3R,GRP75 and VDA C1 mRNA and protein in the injured substantia nigra(P＜0.05,P＜0.01).The high-dose Compound Dihuang Granules presented equivalent efficacy to that of Midoba.CONCLUSION Compound Dihuang Granules may promote endoplasmic reticulum-mitochondria homeostasis,reduce mitochondrial damage and maintain the biological function of dopaminergic neurons by regulating the expressions of DJ-1,IP3R,GRP75 and VDAC1.