Aim To explore the mechanism of rhein in the treatment of IgA nephropathy in rats by impro-ving spleen immune function based on mTORC1 path-way.Methods Thirty-two SD rats were randomly di-vided into control group,model group(IgAN),rhein group(RH)and rapamycin group.The deposition of IgA in mesangial region and pathological changes of spleen and kidney were observed,and the ratio of Th/Ts in spleen was determined by immunohistochemical method.The protein expressions of p-mTOR,p-p70S6k,TGF-β1 and TNF-α in spleen were detected by Western blot.Results Compared with the control group,IgAN group significantly increased urinary sludge red blood cell count,24 hour urinary total pro-tein,serum creatinine,blood urea nitrogen,serum IgA,Th/Ts,p-mtor,p-P70S6K and spleen TNF-αprotein expression levels(P＜0.01),and spleen TGF-β1 protein expression(P＜0.05).IgA deposi-tion in mesangial area of the kidney increased signifi-cantly,and pathological changes were observed in spleen and kidney.Compared with IgAN group,urine sludge red blood cell count,serum IgA,Th/Ts,p-mTOR,p-P70S6K and TNF-α protein expression lev-els in RH and RAPA groups were significantly reduced(P＜0.01).24 hour urinary total protein,blood urea nitrogen,serum creatinine and spleen TGF-β1 protein decreased(P＜0.05),the deposition of IgA in the mesangial region of the kidney decreased,and the pathological changes of spleen and kidney were allevia-ted.There was no significant difference between RH group and RAPA group.Conclusions RH can allevi-ate IgA nephropathy by inhibiting the activity of m-TORC1 pathway in the spleen of IgA nephropathy rats,thereby inhibiting the differentiation of splenic lympho-cytes and reducing the secretion of IgA.

Aim To investigate the effects of dimethyl-arginine dimethylamino hydrolase 1(DDAH1)on high glucose-induced mitochondrial dysfunction and mitoph-agy in vascular endothelial cells.Methods JC-1 stai-ning was used to detect mitochondrial membrane poten-tial.DCFH-DA fluorescent probe was employed to measure reactive oxygen species(ROS)levels.Ho-echst staining was used to assess cell apoptosis.Real-time PCR was conducted to detect DDAH1 mRNA lev-els.Western blot was performed to analyze the expres-sion of DDAH1,LC3-Ⅰ and LC3-Ⅱ proteins.Mitochon-drial probe Mitotracker and autophagosome marker pro-tein LC3 were used in cell immunofluorescence co-lo-calization to assess mitochondrial autophagy,and high-performance liquid chromatography was utilized to measure the levels of asymmetric dimethylarginine(ADMA)in cell supernatant.Results High glucose treatment for 48 h significantly reduced mitochondrial membrane potential,increased ROS production,and promoted apoptosis in human umbilical vein endothelial cells(HUVECs).High glucose downregulated the ex-pression of LC3-Ⅱ/LC3-Ⅰ proteins,reduced the co-lo-calization of Mitotracker and LC3,and inhibited mito-chondrial autophagy.Autophagy inhibitors 3-MA or CQ exacerbated high glucose-induced mitochondrial dam-age and apoptosis in HUVECs,while autophagy activa-tor RAPA alleviated these effects.High glucose signifi-cantly downregulated DDAH1 protein expression in HUVECs and increased ADMA levels in cell superna-tant.DDAH1 siRNA inhibited mitochondrial autoph-agy,reduced mitochondrial membrane potential,and promoted apoptosis,whereas DDAH1 overexpression enhanced mitochondrial autophagy and alleviated high glucose-induced apoptosis in HUVECs.Conclusion High glucose-induced endothelial mitochondrial dys-function is associated with the suppression of DDAH1 expression,the increase of ADMA levels,and thereduc-tion of mitochondrial autophagy.

Coronary artery calcification often appears a variety of complex lesions,increasing coronary intervention of the difficulty of treatment,especially the severe calcification lesions,usually cannot be fully dilated,resulting in a reduced success rate of surgery,an increased rate of acute stent thrombosis and restenosis,and even a serious impact on the prognosis of patients.Intravascular lithotripsy(IVL)is increasingly used in calcified lesions.There is more and more evidence of using in stable angina pectoris and unstable angina pectoris,but its use in acute ST-segment elevation myocardial infarction is limited,and only a few cases have been reported abroad.Moreover,the consensus of Chinese experts in the diagnosis and treatment of coronary artery calcification in 2021 edition lists thrombotic lesions as contraindications of shock wave balloon.This case is the first time in China to report the use of shock wave balloon in patients with acute ST elevation myocardial infarction complicated with thrombus.In this case,the patient with acute ST elevation myocardial infarction complicated with thrombus was severely under expanded stent after stent implantation,and obtain good curative effect using shockwave balloon at selected time in hospital after intensive anticoagulant therapy.

Transfer-RNA derived small RNA(tsRNA),a re-cently discovered class of non-coding RNA,is produced by ma-ture tRNA or tRNA precursor through the mediation of specific endonucleases.By regulating gene expression at the transcrip-tional and post transcriptional levels and acting as an epigenetic regulator,tsRNA plays an important role in the physiological and pathological processes of many organisms.Therefore,it has gradually become a research hotspot in biomedicine and attracted widespread attention.Moreover,there is increasing evidence that tsRNA is involved in the occurrence and development of many neuropsychiatric diseases through participating in stress re-sponse,cell proliferation and apoptosis,neural development,synaptic plasticity,neuroinflammation and immune regulation,epigenetic regulation,RNA processing,and protein translation regulation.This article mainly discusses the generation,classifi-cation and biological functions of tsRNA,and elaborates on the role and possible mechanisms of tsRNA in neurodevelopment and neuropsychiatric disorders,thereby further revealing the poten-tial of tsRNA as a reliable biomarker and therapeutic target for neuropsychiatric disorders.

AIM To investigate the effects of Wuzi Yanzong Pills on subacute aging-induced testicular trauma and AMPK/mTOR pathway in rats.METHODS Rat models of subacute aging were induced by 8-week subcutaneous injection of D-gal(200 mg/kg)into the rat neck,followed by their random assignment into the model group,the metformin group(0.3 g/kg)and the low-dose and high-dose Wuzi Yanzong Pills groups(0.54,2.16 g/kg),with 9 rats in each group,in contrast to the 8 intact rats of the normal group.And the corresponding drug was given by gavage for 4 weeks after modeling,after which the rats had their levels of serum T,FSH,SOD,LH,8-OHdG and MDA detected by ELISA;their pathological changes of testes observed by HE and β-galactosidase staining;and their expressions of aging-related proteins(p16,p21,p53),testicle secretory function-related proteins(CYP11A1,HSD17B3,STAR),autophagy-related proteins(p62,ATG5,Beclin-1,LC3B)and AMPK/mTOR pathway-related protein detected by Western blot and immunohistochemistry.RESULTS Compared with the normal group,the model group displayed morphologically smaller testes and body surface;dry hair;overall atrophic testicular tissue structure;increased testicular protein expressions of γ-H2AX,p16,p21 andβ-galactosidase(P＜0.05,P＜0.01);and decreased p53 protein expression(P＜0.05);suggesting the modeling success.Compared with the model group,the high-dose Wuzi Yanzong Pills group shared decreased levels of serum 8-OHdG,MDA,FSH and LH(P＜0.01);increased levels of SOD and T(P＜0.01);improved sperm damage and almost morphologically normal testicular tissue;decreased testicular protein expressions of p16,p21,p53,p62 and p-mTOR(P＜0.01);and increased protein expressions of CYP11A1,HSD17B3,STAR,ATG5,Beclin-1,LC3B and p-AMPK(P＜0.01).CONCLUSION Wuzi Yanzong Pills can improve the subacute aging-induced testicular injury and testicular function in rats by reducing their oxidative stress through improving their autophagy level and testicle antioxidant capacity.

AIM To study the effects of total flavonoids of Dracocephalum Moldavica L.(TFDM)on reducing the inflammatory response of RAW264.7 macrophages induced by ox-LDL via the nuclear factor κB(NF-κB)/NOD-like receptor 3(NLRP3)signaling pathway.METHODS The RAW264.7 macrophages cultured in vitro were divided into the normal group,the model group(50 μg/mL ox-LDL),the TFDM group(100 μg/mL TFDM+50 μg/mL ox-LDL),the NF-κB inhibitor group(10 μmol/L Bay11-7821+50 μg/mL ox-LDL)and the TFDM+NF-κB inhibitor group(100 μg/mL TFDM+10 μmol/L Bay11-7821+50 μg/mL ox-LDL).The cells had their viability assessed by CCK-8 method;their ROS expression detected by the ROS kit;their mRNA expressions of NF-κB p65,NLRP3,Caspase-1,IL-18 and IL-1β detected by RT-qPCR;their protein expressions of NF-κB p65,IκBα,NLRP3,pro-Caspase-1,Caspase-1,IL-18 and IL-1β by Western blot;their protein expressions of NF-κB p65 and NLRP3 detected using immunofluorescence method.RESULTS Compared with the normal group,the model group showed increased ROS expression(P＜0.01);increased mRNA expressions of NF-κB p65,NLRP3,Caspase-1,IL-18 and IL-1β(P＜0.05,P＜0.01);decreased protein expressions of IκBα and cytoplasmic NF-κB p65(P＜0.01);increased protein expressions of nuclear NF-κB p65,NLRP3,Caspase-1,IL-1 β and IL-18(P＜0.01);and increased fluorescence intensity of NF-κB p65 and NLRP3(P＜0.01).Compared with the model group,the groups intervened with either TFDM or TFDM+inhibitor displayed decreased ROS expression(P＜0.01);the groups administrated with TFDM or NF-κB inhibitor,or TFDM+inhibitor showed decreased mRNA expressions of NF-κB p65,NLRP3,Caspase-1,IL-18 and IL-1β(P＜0.05,P＜0.01),increased protein expressions of IκBα and cytoplasmic NF-κB p65(P＜0.05,P＜0.01),decreased protein expressions of nuclear NF-κB p65,NLRP3,Caspase-1,IL-1β and IL-18(P＜0.05,P＜0.01),and decreased fluorescence intensity of NF-κB p65 and NLRP3(P＜0.01).There existed no significant group difference between the TFDM group and the NF-κB inhibitor group(P＞0.05).The TFDM+inhibitor group demonstrated decreased mRNA expressions of IL-1βand IL-18(P＜0.05),increased IκBα protein expression(P＜0.05),decreased protein expressions of nuclear NF-κB p65,NLRP3,Caspase-1,IL-1 β and IL-18(P＜0.05),and decreased fluorescence intensity of NLRP3 protein(P＜0.05).CONCLUSION TFDM can inhibit the ox-LDL-induced inflammatory response of RAW264.7 macrophages,and the mechansism may be associated with the reduced ROS expression and inflammatory factors due to the inhibited activation of the NF-κB/NLRP3 signaling pathway.

To investigate the anti-atherosclerotic effects of dietary hesperidin and naringin on ather-osclerosis in rabbits.Twenty-four adult healthy male New Zealand Large White rabbits were ran-domly assigned to four groups based on one factor:control(Con),model(Mod),hesperidin(HP),and naringenin(NG),each containing six rabbits and housed in solitary cages.During the experi-mental period,the control group received normal feed;the model group was fed high-fat for 30 d to establish the atherosclerosis,(AS)model and continued to be fed high-fat feed;the hesperidin and naringenin groups were fed high-fat for 30 d and then underwent AS modeling,and at a later stage,150 mg of hesperidin(or naringenin)was added to the diets at a rate of 1.5 mg per kg of di-et,respectively,for 30 d.Relative body quality,blood lipids,oxidation,and inflammatory factor lev-els were all assessed.At the end of the test period,the test rabbits were sacrificed to obtain the common carotid artery for histopathological section HE staining,Oil red O staining,and Sirius red staining in order to observe vascular tissue structure,lipid and inflammatory cell infiltration,and e-lastic plate and elastic fiber damage.The results indicated that the relative body quality of the mod-el,HP and NG groups increased rapidly,and the difference with the control group within the same period was highly significant(P＜0.01);The HP and NG interventions significantly decreased to-tal cholesterol(TC)and low-density lipoprotein cholesterol(LDL-C)levels(P＜0.01),while also leading to a significant increase in high-density lipoprotein cholesterol(HDL-C)levels(P＜0.01);MDA levels exhibited a significant decrease(P＜0.01),while SOD levels showed a notable in-crease(P＜0.01)following the HP and NG interventions;After HP and NG interventions,TNF-α and IL-1 were regressed(P＜0.01)and IL-10 was significantly elevated(P＜0.01);Compared with the control group,the model group showed approximately 41.8 and 2.5 fold thickening of the maximal endo-medial membrane,respectively,and the HP and NG interventions showed approxi-mately 13 and 15 fold thickening of the maximal endo-medial membrane,and both of the maximal medial membranes thickened by approximately 1.4 fold;Staining of pathological sections showed that HP and NG intervened to significantly reduce inflammatory cell infiltration,deposition of lipid components,slight thickening of the endothelium,and intact fibroblastic components.The results showed that the addition of HP and NG to diets could regulate blood lipids and exert anti-inflam-matory and antioxidant effects by inhibiting the expression of inflammatory factors and oxidizing factors,showing significant anti-AS effects,and the two effects were comparable.

Objective To investigate the effects of ischemia time and storage periods on RNA quality in fresh-frozen breast cancer(BC)and esophageal cancer(EC)tissue samples in order to establish evidence-based protocols for biobank sample management.Methods The tumor(T)and paired normal(N)tissue samples from 6 cases of BC and 6 cases of EC were collected and cryopreserved in Biobank,Shantou Central Hospital.Mirror paraffin-embedded tissues were simultaneously prepared into sections for morphological analysis.The samples were divided into two groups of<15 min and 15-30 min according to ischemia time,and RNA quality was analyzed at 4 storage periods of 8-10 months(T1),14-16 months(T2),26-28 months(T3)and 38-40 months(T4).Results In 96 analyzed samples,93.8%(90/96)exhibited high quality(RIN≥6),with 89.6%(43/48)in BC and 97.9%(47/48)in EC.Significant differences in RIN were observed between BC group and EC group(8.050 vs.8.600,P=0.009).In EC group,RIN value was significantly negatively correlated with RNA yield(P<0.001).Moreover,RIN values of tumor-normal pairs exhibited markedly significant differences(7.550 vs.9.000,P<0.001).In contrast,no significant difference was detected in BC group(8.200 vs.7.700,P=0.348).Statistical analysis showed that RIN value was positively correlated with 28S/18S(P<0.001),but had no correlation with tumor content(P=0.676)and necrotic content(P=0.055).Neither ischemia time(<15 min vs.15-30 min:8.200 vs.8.300,P=0.932)nor storage periods(T1-T4:8.400,7.700,8.450,8.600,P=0.163)compromised RNA quality.Conclusion Organ origin and tissue type could influence RNA quality of fresh-frozen tissue samples.However,limited ischemia time(≤30 min)and long-term storage period(38-40 months)do not adversely affect RNA quality in fresh-frozen breast cancer and esophageal cancer tissue samples.

In view of the characteristics of H5N6 subtype avian influenza virus(AIV)that it has both high pathogenicity and the risk of cross-species transmission,posing a serious threat to the poultry farming industry and public health security,in order to effectively prevent and control the spread of H5N6 avian influenza,a rapid,sensitive and specific detection technolo-gy was established in this study.The specific monoclonal antibodies against the neuraminidase N6 protein of avian influenza A virus subtype H5N6 were obtained through hybridoma and monoclonal antibody technology.These antibodies were coupled and labeled with carboxyl-functionalized fluorescent quantum dots,along with previously prepared specific antibodies against the hemagglutinin H5 protein.A rapid fluorescence immunochromatographic detection method for the H5N6 subtype of avian influ-enza virus was established according to the principle of double-antibody sandwich immunochromatography.This method a-chieved a detection sensitivity of 1 ng/mL for recombinant hemagglutinin H5 subtype protein and 0.1 ng/mL for recombinant neuraminidase N6 subtype protein.Moreover,the method exhibited no cross-reactivity with other influenza subtypes or patho-gens,such as Newcastle disease(ND),infectious bronchitis(IB),and infectious laryngotracheitis(ILT),thus demonstrating good specificity.The method effectively identified the highly pathogenic avian influenza virus H5 subtype and directly distin-guished the H5N6 subtype with good accuracy.The fluorescent quantum dot immunochromatographic typing detection method established herein met the sensitivity,specificity,and accuracy requirements for H5N6 subtype detection,and can be further used for rapid detection of the H5 and H5N6 subtypes of avian influenza virus.

Aim To explore the mechanism of rhein in the treatment of IgA nephropathy in rats by impro-ving spleen immune function based on mTORC1 path-way.Methods Thirty-two SD rats were randomly di-vided into control group,model group(IgAN),rhein group(RH)and rapamycin group.The deposition of IgA in mesangial region and pathological changes of spleen and kidney were observed,and the ratio of Th/Ts in spleen was determined by immunohistochemical method.The protein expressions of p-mTOR,p-p70S6k,TGF-β1 and TNF-α in spleen were detected by Western blot.Results Compared with the control group,IgAN group significantly increased urinary sludge red blood cell count,24 hour urinary total pro-tein,serum creatinine,blood urea nitrogen,serum IgA,Th/Ts,p-mtor,p-P70S6K and spleen TNF-αprotein expression levels(P＜0.01),and spleen TGF-β1 protein expression(P＜0.05).IgA deposi-tion in mesangial area of the kidney increased signifi-cantly,and pathological changes were observed in spleen and kidney.Compared with IgAN group,urine sludge red blood cell count,serum IgA,Th/Ts,p-mTOR,p-P70S6K and TNF-α protein expression lev-els in RH and RAPA groups were significantly reduced(P＜0.01).24 hour urinary total protein,blood urea nitrogen,serum creatinine and spleen TGF-β1 protein decreased(P＜0.05),the deposition of IgA in the mesangial region of the kidney decreased,and the pathological changes of spleen and kidney were allevia-ted.There was no significant difference between RH group and RAPA group.Conclusions RH can allevi-ate IgA nephropathy by inhibiting the activity of m-TORC1 pathway in the spleen of IgA nephropathy rats,thereby inhibiting the differentiation of splenic lympho-cytes and reducing the secretion of IgA.