ObjectiveTo investigate the potential mechanisms of Zhimu （Anemarrhena asphodeloides）-Huangbai （Phellodendron amurense） medicinal pair in alleviating postmenopausal osteoporosis （PMOP）. MethodsSixty unpregnant female SD rats were randomly divided into five groups， blank group， model group， low-dose Zhimu-Huangbai group， high-dose Zhimu-Huangbai group， and estradiol group， with 12 rats in each group. Except for the blank group， all other groups had their ovaries removed to create PMOP rat models， while the blank group only had the fat tissue around the ovaries removed. One week after the ovarian removal， the low-dose and high-dose Zhimu-Huangbai groups received concentrated solution of Zhimu and Huangbai with 1.8， 7.2 g/（kg·d） via gavage， the estradiol group received estradiol solution 0.09 mg/（kg·d） via gavage， and the blank group and the model group received 10 ml/（kg·d） of normal saline via gavage， once daily for 12 weeks. Before sampling， the body mass of the rats was recorded， and uterine tissue was taken to calculate the uterine index. The levels of interleukin-1β （IL-1β）， interleukin-6 （IL-6） and tumor necrosis factor-α （TNF-α） in serum were detected by ELISA； micro CT was used to examine the parameters of femoral microstructure， including bone volume/tissue volume （BV/TV）， trabecular number （Tb.N）， trabecular thickness （Tb.Th）， bone mineral density （BMD）， trabecular separation （Tb.Sp）， and cortical bone area （Ct.Ar）. HE staining was used to observe pathological changes in the femur； RT-qPCR was used to detect the mRNA expression of osteogenic-adipogenic differentiation-related factors in femoral tissue， including Runt-related transcription factor 2 （Runx2）， bone morphogenetic protein 2 （BMP-2）， peroxisome proliferator-activated receptor γ （PPARγ）， chemerin and chemokine-like receptor 1 （CMKLR1）. ResultsCompared with the blank group， the model group showed a significant increase in body mass， a significant decrease in the uterine index， BV/TV， Tb.N， Tb.Th and BMD， a significant increase in Tb.Sp， and serum IL-1β， IL-6， and TNF-α levels， a significant reduction of mRNA expression of Runx2 and BMP-2 in bone tissue， and a significant increased mRNA expression of PPARγ， chemerin， and CMKLR1 （P＜0.01）. HE staining revealed that the femoral tissue showed a reduction and sparsity of trabeculae， a significant enlargement of the medullary cavity， and a large number of fat cells. Compared to the model group， the low-dose， high-dose Zhimu-Huangbai groups， and estradiol group showed significant improvements in all the above-mentioned indicators （P＜0.05 or P＜0.01）. HE staining revealed a significant increase in trabeculae， more organized arrangement， and a marked reduction in fat cells. Compared to low-dose Zhimu-Huangbai group， the high-dose Zhimu-Huangbai group exhibited a significant increase in the uterine index and BMD， and a significant reduction in body mass and PPARγ and Chemerin mRNA expression （P＜0.05 or P＜0.01）. Compared to high-dose Zhimu-Huangbai group， the estradiol group showed a decrease in uterine index， BV/TV， Tb.N， Tb.Th， BMD， and BMP-2 mRNA expression， while the levels of IL-1β， TNF-α， and IL-6， as well as Tb.Sp and the mRNA expressions of PPARγ， chemerin， and CMKLR1 increased （P＜0.05 or P＜0.01）. ConclusionThe Zhimu-Huangbai medicinal pair can alleviate PMOP bone loss， and its mechanism of action is related to reducing the levels of inflammatory factors， correcting the disorder of osteogenic-adipogenic differentiation of bone marrow mesenchymal stem cells （BMSCs）， and promoting the differentiation of BMSCs into osteoblasts.

Objective To investigate and evaluate the nutrition status in serum 25-hydroxy vitamin D[25(OH)D]levels of 0～12 years old children in Xi'an.Methods A total of 2 670 patients aged 0～12 years old who underwent routine physical examinations in Children's Health Department of the Second Hospital of Air Force Medical University were selected from March 2020 to July 2023,and the 25(OH)D data of these patients were conducted in retrospective analysis.The nutritional status of vitamin D in these children of different genders,ages and seasons were also analyzed.Results ①This study included 2 670 children aged 0～12 years old in Xi'an,with the average level of serum 25(OH)D was 40.80±18.00 ng/ml.Among them,38 cases(1.42%)had serum 25(OH)D deficiency,333 cases(12.47%)had serum 25(OH)D insufficience,and 2 299 cases(86.11%)had sufficient serum 25(OH)D.②There was a statistically significant difference in serum 25(OH)D levels among different age groups(H=1 524.23,P=0.000).The group aged 1～<4 has the highest value of 52.51±13.57 ng/ml,while the group aged 8～12 has the lowest value of 21.65±6.75 ng/ml.③The levels of serum 25(OH)D in summer(39.44±17.46 ng/ml)were lower than those in spring(41.96±17.76 ng/ml)and autumn(42.71±18.15 ng/ml),with statistical significant differences(Z=101.57,-134.06,all P<0.01).However,the deficiency and inadequate rate of serum 25(OH)D in winter(18.95%)was higher than those in spring,summer and autumn(13.52%,12.75%,12.36%),with statistical significant differences(χ2=14.32,P=0.026).④There was no statistically significant difference in serum 25(OH)D levels between genders(H=0.933,P=0.351).However,the deficiency and inadequate rate of serum 25(OH)D in boys(12.51%)was lower than that in girls(15.46%),and the difference was statistically significant(χ2=9.257,P=0.010).Conclusion The nutritional status of serum 25(OH)D in children aged 0～12 years in Xi'an area is comparatively fine,and it is necessary to strengthen the intake and supplementation of vitamin D in over 3 years old children.

Objective:To explore the value of applying multiple genetic testing techniques for the prenatal diagnosis of Turner syndrome fetuses with complex mosaic small supernumerary marker chromosomes (sSMC).Methods:Chromosomal karyotypes of amniotic fluid samples from 5 030 pregnant women who had undergone amniocentesis at the Prenatal Diagnosis Center of the Third Affiliated Hospital of Zhengzhou University from January to December 2022 were retrospectively reviewed. Three fetuses with complex mosaicism fetuses (carrying 2 types of sSMC) were selected as the study subjects. Genetic tests including G-banded chromosomal karyotyping analysis, fluorescence in situ hybridization (FISH), chromosomal microarray analysis (CMA), and copy number variation sequencing (CNV-seq) were used to clarify the origin and mosaic status of the sSMC. This study has been approved by the Medical Ethics Committee of the Third Affiliated Hospital of Zhengzhou University (No. 2023-159-01).Results:G-banded chromosomal analysis of fetus 1 showed a karyotype of 45, X[64]/46, X, + mar1[13]/46, X, + mar2[3]. FISH results showed that 52% of of its cells had contained one X chromosome signal, whilst 48% contained two X chromosome signals. CMA results revealed the fetus had harbored a 32.32 Mb and a 50.93 Mb deletion in Xp22.33p21.1 and Xq22.2q28 regions, respectively, in addition with mosaic deletions of approximately 1.43 copies, 1.78 copies and 1.43 copies in the Xp21.1p11.1, Xq11.1q21.1 and Xq21.2q22.2 regions, respectively. The fetus 2 had a karyotype of 45, X[27]/46, X, + mar1[14]/46, X, + mar2[12]. FISH results indicated that 88% of its cells contained one X chromosomes signal and two Y chromosome signals, and 12% contained signals for one X chromosomes signal and one Y chromosome signal. CNV-seq results revealed a deletion of 7.74 Mb in the Yq11.222q11.23 region and a mosaic duplication of approximately 1.738 copies in the Yp11.31q11.221 region. The fetus 3 had a karyotype of 45, X[60]/46, X, + mar1[11]/46, X, + mar2[6]. FISH results showed that 28% of its cells contained one X chromosome signal, and 72% contained tow X chromosome signals. CNV-seq results revealed deletions of 55.60 Mb and 53.50 Mb in the Xp22.33p11.1 and Xq22.1q28 regions, respectively, along with a mosaic deletion of approximately 1.85 copies in the Xp11.1q13.2 region and a mosaic repeats of approximately 2.66 copies in the Xq13.2q22.1 region. The sSMCs in the 3 fetuses had all originated from sex chromosomes and were of complex mosaic type. After genetic counseling, the three couples had all opted to terminate the pregnancy.Conclusion:The combined use of multiple genetic testing techniques has determined the origin and structure of complex mosaic sSMCs and provided a basis for prenantal diagnosis and genetic counseling.

Tetrandrine(TET),a natural bisbenzyl isoquinoline alkaloid extracted from Stephania tetrandra S.Moore,has diverse pharmacological effects.However,its effects on melanoma remain unclear.Cellular prolif-eration assays,multi-omics analyses,and xenograft models were used to determine the effect of TET on melanoma.The direct target of TET was identified using biotin-TET pull-down liquid chromatograph-mass spectrometry(LC-MS),cellular thermal shift assays,and isothermal titration calorimetry(ITC)analysis.Our findings revealed that TET treatment induced robust cellular autophagy depending on activating transcription factor 6(ATF6)-mediated endoplasmic reticulum(ER)stress.Simultaneously,it hindered autophagic flux by inducing cytoskeletal protein depolymerization in melanoma cells.TET treatment resulted in excessive accumulation of reactive oxygen species(ROS)and simultaneously triggered mitophagy.Sirtuin 5(SIRT5)was ultimately found to be a direct target of TET.Mechanistically,TET led to the degradation of SIRT5 via the ubiquitin(Ub)-26S proteasome system.SIRT5 knockdown induced ROS accumulation,whereas SIRT5 overexpression attenuated the TET-induced ROS accumula-tion and autophagy.Importantly,TET exhibited anti-cancer effects in xenograft models depending on SIRT5 expression.This study highlights the potential of TET as an antimelanoma agent that targets SIRT5.These findings provide a promising avenue for the use of TET in melanoma treatment and underscore its potential as a therapeutic candidate.

AIM:To investigate the effects of Shaoyao-Gancao decoction(SGD)on inflammation and mucosal barrier damage in rats with 2,4,6-trinitrobenzenesulfonic acid(TNBS)-induced inflammatory bowel disease(IBD).METHODS:Forty-eight male SD rats were randomly divided into normal group,model group,high-dose(500 mg/kg),medium-dose(250 mg/kg)and low-dose(125 mg/kg)SGD groups,and balsalazide sodium(1 g/kg)group.All rats were pre-administered for 3 d,and on the 4th day of the experiment,they were fasted for 24 h.Except for the normal group,the rats in the other 5 groups were given enema mixed with TNBS(100 mg/kg)and 50％ethanol,and continued to be adminis-tered for 5 d after modeling.After modeling,the disease activity index(DAI)was evaluated.After the experiment,the levels of nitric oxide(NO)and myeloperoxidase(MPO)in serum and colonic tissues of rats were determined.RT-qPCR and Western blot were used to determine tumor necrosis factor-α(TNF-α),cyclooxygenase-2(COX-2),inducible nitric oxide synthase(iNOS)and nuclear factor-κB(NF-κB)in the colon of rats.The expression of tight junction proteins zonu-la occludens-1(ZO-1)and claudin 2 in rat colon tissues was determined by immunofluorescence staining.RESULTS:Compared with normal group,the weight of rats in model group was decreased,the colon was shortened,DAI and colon tissue macroscopic scores were significantly increased(P<0.05),colon pathological injury was serious,and NO and MPO levels in serum and colon tissues of the rats in model group were significantly increased(P<0.05).The mRNA and pro-tein expression levels of TNF-α,COX-2,iNOS and NF-κB in colon tissues were significantly increased(P<0.01),while the expression levels of ZO-1 and claudin 2 were significantly decreased(P<0.01).Compared with model group,the body weight and colon shortening of rats in SGD groups were alleviated,DAI and macroscopic scores of colon tissues were significantly decreased(P<0.05),the pathological damage of colon was improved,and the levels of NO and MPO in se-rum and colon tissues of rats were significantly decreased(P<0.05).The mRNA and protein expression levels of TNF-α,COX-2,iNOS and NF-κB in colon tissues were significantly decreased(P<0.05),while the expression levels of ZO-1 and claudin 2 were significantly increased(P<0.05).CONCLUSION:Treatment with SGD effectively attenuates the inflam-matory response and intestinal mucosal barrier damage caused by TNBS-induced IBD in rats.

UNC-51-like kinase 1(ULK1)is an important factor involved in regulating the initiation of autophagy.ULK1 regu-lates inflammatory cytokines through autophagy and mitochondrial oxidative stress,and is involved in the pathological processes of var-ious diseases.ULK1 and its complexes are regulated by rapamycin(mTOR)and AMP-activated protein kinase(AMPK)to initiate au-tophagy,thereby exerting differential effects on a variety of inflammatory diseases.In inflammatory diseases,mitochondrial oxidative stress can induce ULK1 into the nucleus to accelerate apoptosis.Therefore,ULK1 plays different important roles in inflammatory dis-eases.For example,ULK1 initiates airway epithelial mitochondrial autophagy in asthma,participates in mitochondrial oxidative stress in acute liver failure,affects related inflammatory factors in atherosclerosis,and modulates beneficial effects of autophagy in diabetes.This article reviews the biological function of ULK1,its impact on inflammatory diseases and the research progress of targeted drugs.

Rabies is a zoonotic disease that poses a global public health threat.Rabies virus(RABV)is neurotropic and can cause severe neurological disorders and behavioral abnormalities in host,with a fatality rate nearly 100％.In order to identify the key genes for RABV affecting neuronal cell function,we established and analyzed the mRNA expression profile of Neuro-2a(N2a)cell in-fected with challenge virus standard(CVS)-11 by RNA sequencing(RNA-seq).Biological func-tions of differentially expressed genes(DEGs)were determined by GO and KEGG enrichment a-nalysis.The results showed that there were 415 differentially expressed genes in N2a infected with CVS-11 strain,of which 89 were up-regulated and 326 were down-regulated.These genes were re-lated to a variety of biological processes,such as axon guidance pathway,cholesterol metabolism pathway,nitrogen metabolism pathway,and glycosphingolipid biosynthesis pathway,many of them have been shown to be closely associated with RABV infection.A total of 12 DEGs related to axon conduction,antigen processing and presentation pathways were selected and detected by real-time PCR,and their expression trends were consistent with the RNA-seq results.The genomic tran-scriptomic data on N2a cell under RABV infection will provide new clues for probing the mecha-nisms of RABV infection and transmission in the nervous system in the future.

Objective:To investigate the relationship between non-alcoholic fatty liver disease(NAFLD) and hepatic fibrosis and skeletal muscle mass in patients with type 2 diabetes mellitus(T2DM).Methods:A total of 685 T2DM patients diagnosed at the Endocrinology department of Lanzhou University First Hospital, from April 2022 to May 2023, were divided into NAFLD and Non-NAFLD groups, and the NAFLD group was further categorized into fibrosis and non-fibrosis based on aspartate aminotransferase(AST) /alanine aminotransferase(ALT) level. The differences in appendicular skeletal muscle mass(ASM), appendicular skeletal muscle mass index(ASMI), and the prevalence of muscle mass loss were compared across groups. The correlations between ASMI and NAFLD, as well as liver fibrosis were analyzed by binary logistic regression.Results:Among male T2DM patients, those with NAFLD had lower ASMI levels and a higher prevalence of muscle mass reduction compared to non-NAFLD group. Among female T2DM patients, those with NAFLD had lower levels of ASM and ASMI, and a higher prevalence of muscle mass reduction compared to non-NAFLD group. ASMI levels in both male and female T2DM patients were independently negatively correlated with NAFLD risk( OR=-0.696, 95% CI 0.579-0.837; OR=-0.757, 95% CI 0.629-0.911). In NAFLD patients, ASM and ASMI levels were lower in those with liver fibrosis compared to those without fibrosis; however, the prevalence of muscle mass reduction did not differ significantly. Among male NAFLD patients, ASMI levels were independently negatively correlated with the risk of liver fibrosis( OR=-0.726, 95% CI 0.537-0.983), while no correlation was found in female patients. Conclusion:Reduced muscle mass is independently associated with the risk of NAFLD in both male and female T2DM patients. In males, reduced muscle mass is also independently related to the risk of liver fibrosis.

Objective To investigate the neuroprotective effect and mechanism of salvia polyphenolic acid for injection (SAFI) on cerebral ischemia-reperfusion injury in rats. Methods A total of 100 SD rats were randomly divided into sham surgery group,model group,low-,medium-and high-dose (5,10,20 mg·kg-1) salvia polyphenolic acid groups,with 20 rats in each group. After being continuously administrated by intraperitoneal injection of SAFI once daily for three days,the rat model of middle cerebral artery occlusion/reperfusion (MCAO/R) was established using the thread embolization method at 1 hour after the last administration. The neurological deficit of rats was evaluated by Zea Longa score. The cerebral infarction volume was detected by 2,3,5-triphenyltetrazolium chloride(TTC) staining. The levels of serum NADPH oxidase(NOX),4-hydroxynonanal(4-HNE),8-hydroxydeoxyguanosine (8-OHdG),monocyte chemoattractant protein-1 (MCP-1),tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β),interleukin-18(IL-18),interleukin-6(IL-6),and intercellular adhesion molecule-1 (ICAM-1) were detected by ELISA kits. Hematoxylin-eosin (HE) staining and Nissl staining were used to observe the pathological changes of brain tissue and the morphology of neurons. The apoptosis of neuronal cells in brain tissue was detected by TUNEL. Immunofluorescence staining was used to detect the expression level of glial fibrillary acidic protein (GFAP) in brain tissue. Western Blot was used to detect the protein expression of NLRP3 and Caspase1 in brain tissue. Results Compared with the sham surgery group,neurological deficit scores in model group increased remarkably (P<0.01). The cerebral infarction volume increased significantly (P<0.01). Serious pathological damage of brain was observed,and neuronal density decreased significantly(P<0.01). The apoptosis rate of cortical cells increased obviously (P<0.01). The levels of serum NOX,4-HNE,8-OHdG,MCP-1,TNF-α,IL-1β,IL-18,IL-6 and ICAM-1 increased significantly (P<0.05,P<0.01). The protein expression of GFAP,NLRP3 and Caspase1 in brain significantly upregulated (P<0.01). Compared with the model group,neurological deficit scores in medium-and high-dose SAFI groups decreased remarkably (P<0.01). The cerebral infarction volume decreased significantly (P<0.01). Neuronal damage was ameliorated to varying degrees,and neuronal density increased significantly(P<0.05,P<0.01). The apoptosis rate of cortical cells decreased obviously (P<0.01). The levels of serum NOX,4-HNE,8-OHdG,MCP-1,TNF-α,IL-1β,IL-18,IL-6 and ICAM-1 decreased significantly (P<0.05,P<0.01). The protein expression of GFAP,NLRP3 and Caspase1 in brain significantly downregulated(P<0.01). Conclusion SAFI has a protective effect on MCAO/R rats,which can significantly reduce oxidative stress and inflammatory responses,thereby reducing pathological damage and apoptosis of brain tissue. Its mechanism may be related to the inhibition of NLRP3/Caspase-1 signaling pathway and astrocyte activation.

Hemophagocytic syndrome (HPS), which is currently named as hemophagocytic lymphohistiocytosis (HLH), is a hyperinflammatory syndrome characterized by persistent fever, hepatosplenomegaly, pancytopenia and hemophagocytosis found in bone marrow, liver, spleen and lymph nodes due to excessive activation of macrophages and cytotoxic T cells. Macrophage activation syndrome (MAS) is a specific form of HLH induced by autoinflammatory/autoimmune disorders which can be life-threatening and requires multiple disciplines. In order to improve clinicians′ understanding of MAS and standardize the clinical diagnosis and treatment practice of MAS, the rheumatology branch of Chinese Rheumatology Association organized domestic experts to formulate the diagnosis and treatment standard, in order to improve the diagnosis and treatment level of MAS and improve the prognosis of patients.