Traditional Chinese medicine （TCM） has long recognized metabolism-associated fatty liver disease （MASLD）. The Classic of Difficulties （Nan Jing） records that "the accumulation of the liver is called obese Qi". ZHANG Jingyue also stated， "People with spleen and stomach deficiency and weakness or imbalance often suffer from diseases of accumulation". According to syndrome differentiation of the Zang-fu organs， modern physicians generally believe that the key pathogenesis of MASLD lies in the deficiency of spleen and stomach functions. However， MASLD is a chronic and complex disease， and its pathological characteristics cannot be fully explained by a single Zang-Fu syndrome differentiation. The concepts of sweat pores and collateral vessels emerged as early as the Huangdi's Internal Classic （Huang Di Nei Jing）， and later TCM scholars， based on the idea that "sweat pore is the gateway of collateral vessels" proposed the concept of Xuanluo （sweat pores-collateral vessels）. Xuanluo refers to fine structures widely distributed throughout the human body， serving as hubs and channels that regulate the movement and distribution of Qi， blood， and body fluids. By linking the Zang-Fu organs with Xuanluo， a theoretical framework of Qi， blood， and body fluid circulation centered on the "spleen and stomach-Xuanluo" as a whole was established， providing a new perspective for analyzing the essential pathogenesis of MASLD. Combined with the mechanisms involved in the formation and progression of MASLD， the intrinsic correlations between TCM pathogenesis and modern microscopic mechanisms are further analyzed. Modern studies have shown that intestinal microbial dysbiosis and mitochondrial dysfunction are pathological mechanisms involved in the occurrence and development of MASLD， but few have discussed the two as an integrated system. Existing research has confirmed that intestinal microorganisms can affect mitochondrial energy metabolism and oxidative stress through their metabolites， leading to hepatic energy metabolism disorders， oxidative stress （OS）， and inflammation， thereby promoting MASLD progression. Focusing on the correspondence between the "spleen and stomach-Xuanluo" theory and the intestinal microorganism-mitochondrion micro-pathological mechanism， it is proposed that the spleen and stomach share similarities with intestinal microorganisms in the generation of Qi， blood， and body fluids as well as in the regulation of Qi movement， while Xuanluo and mitochondria have commonalities in energy regulation. Moreover， harmonizing the spleen and stomach to ensure unobstructed Xuanluo is the key to maintaining the normal interaction mechanism between intestinal microorganisms and mitochondria. Based on the correlation between the "spleen and stomach-Xuanluo" theory and the intestinal microorganism-mitochondrion interaction， this paper reveals that the essence of MASLD pathogenesis lies in spleen and stomach dysfunction， specifically， failure of the spleen to ascend the clear and failure of the stomach to descend the turbid， resulting in insufficient transformation of Qi and blood， impaired nourishment of Xuanluo， stagnation of Qi and blood， and the long-term formation of phlegm and blood stasis in the liver. Furthermore， it explores the preventive and therapeutic effects of tonifying the spleen and stomach， dredging Xuanluo and collaterals， unblocking the bowels， and regulating Qi in the treatment of MASLD， thereby providing new insights for its prevention and therapy.

Objective: To analyze the association between each regional fat mass and total body bone mineral density (BMD) in children and adolescents aged 7-17 years in Beijing, so as to provide theoretical basis and practical guidance for implementing interventions. Methods: From September to December 2020, a stratified cluster random sampling method was used to select 1 423 children and adolescents aged 7-17 years in Tongzhou District, Beijing. Dual energy X-ray absorptiometry (DXA) was employed to measure regional body composition and total body BMD. Multiple linear regression was used to analyze the association between regional fat mass and total body BMD. Results: The median (interquartile range) fat mass values for total body, upper limbs, abdomen, hips, and thighs were 13.51(8.84, 19.21), 1.59(1.08, 2.23), 0.73(0.39, 1.29), 2.32(1.46, 3.26), 5.29(3.59, 7.21)kg, respectively. After adjusting for covariates, the results of multiple linear regression analysis showed that total body fat mass (β=0.010), abdominal fat mass (β=-0.100), and hip fat mass (β=0.104) were significant associations with total body BMD (all P<0.01). Sexstratified analysis revealed that in boys, total body fat mass (β=0.008) and hip fat mass (β=0.058) were positively associated with BMD, while thigh fat mass (β=-0.038) showed a negative association with total body BMD (all P<0.05). In girls, total body fat mass (β=0.013), hip fat mass (β=0.163), and thigh fat mass (β=0.023) were positively associated with total body BMD, whereas abdominal fat mass (β=-0.196) showed a negative association with total body BMD (all P<0.05). Among children and adolescents with body fat percentage below the standard range, within the standard range and above the standard range, total body fat masses were positively associated with total body BMD (β=0.021, 0.016, 0.015); among children and adolescents with body fat percentage within the standard range while upper limb (β=-0.042), abdominal (β=-0.067), and thigh fat mass (β=-0.018) showed negative associations with total body BMD, and hip fat mass demonstrated a positive association with total body BMD (β=0.082) (all P<0.05). Conclusion Regional fat distribution is associated with total body BMD in children and adolescents, with the nature of these associations varying by sex and body fat percentage.

Objective To explore the effective chemical components of Fuzhenghuaxian decoction and its mechanism of action in the treatment of liver fibrosis based on UPLC-Q-TOF-MS combined with network pharmacology and experimental verification.Methods UPLC-Q-TOF-MS/MS technology was used to analyze the active components of Fuzhenghuaxian decoction.The potential targets and mechanisms of FZHXD were analyzed from the aspects of pharmacokinetics,target information,GO enrichment and KEGG Pathway.FZHXD was used to verify the key pathways and targets screened by network pharmacology in mice with liver fibrosis induced by carbon tetrachloride(CCl4),and further improve the accuracy of its prediction results.Results A total of 35 effective chemical components were identified,mainly saponins and organic acids.Through network pharmacology screening,669 drug targets,440 common targets,5 core targets and 11 key active ingredients were obtained.The main signal transduction pathways were predicted to be PI3K-Akt,FOXO,NF-κB signaling pathway,etc.In vivo experiments confirmed that FZHXD improved liver fibrosis in mice through PI3K/AKT,FOXO and NF-κB signaling pathways.Conclusion Fuzhenghuaxian decoction has the characteristics of multi-pathway and multi-target in anti-liver fibrosis by down-regulating PI3K,AKT,NF-κB and up-regulating FOXO.

Objective To optimize a C57BL/6J mouse liver fibrosis model induced by different doses of carbon tetrachloride through imaging,molecular biology,and pathology method.Methods Thirty-six healthy C57BL/6J male mice were randomly divided into a control group,2 weeks,3 weeks,4 weeks,6 weeks,and 8 weeks groups(n=6)after adaptive feeding for 1 week.The control group was intraperitoneally injected with 0.2 mL olive oil three times a week,and the positive-control groups were intraperitoneally injected with 0.2 mL 20%CCl4-olive oil solution three times a week.Changes in the body weights of mice in each group were recorded.Liver stiffness was measured on days 15,22,29,43 and 57,and blood samples were collected,and cereal third alanine aminotransferase(ALT),aspartate aminotransferase(AST),hyaluronic acid(HA),laminin(LN),pro-typeⅢcollagen(PC-Ⅲ),and typeⅣcollagen(Ⅳ-C)content was measured.The liver tissues were stained with hematoxylin and eosin(HE),Masson,and Sirius red.The Metavir scoring system was used to evaluate the degree of liver fibrosis.Results Compared with the control group,mice in the positive-control groups were listless and tended to huddle together.In terms of body weight,the 4 weeks,6 weeks,and 8 weeks groups were significantly lighter than the control group,while the 2 weeks group mice were significantly heavier than the control group mice.Liver elastography showed a progressive increase in stiffness with increased administration time.The biochemical tests showed that,compared with the control group,the other groups'ALT and AST levels were significantly higher.With an increase in drug delivery time,the positive-control group's HA,LN,PC-Ⅲ and Ⅳ-C levels showed increasing trends.Pathological examination revealed that liver fibrosis was progressively aggravated with an increase in administration time.At 4 weeks,the pathological diagnosis was consistent with that of liver fibrosis,and there were signs of pseudolobule formation at 6 weeks.Pseudolobules were formed at 8 weeks,suggesting early cirrhosis.Conclusions A liver fibrosis model can be successfully established in C57BL/6J mice by intraperitoneal injection of 20%CCl4-olive oil solution three times a week for 4 consecutive weeks.The model has good stability,and the modeling method is rapid and can be used as an optimized scheme for the establishment of liver fibrosis models.

Intracranial germ cell tumors (iGCT) is a common cause of delayed puberty (DP).　A retrospective analysis was conducted on the clinical data of two patients with DP caused by iGCT. The disease characteristics of the two patients included DP, primary amenorrhea, growth retardation, short stature, subclinical hypothyroidism, etc. They were diagnosed as DP through imaging and related biochemical tests, and some sex hormone drugs were given on the basis of active treatment of the primary disease. Two patients had varying degrees of development in height, weight and secondary sexual characteristics, and further treatment condition is being tracking by us. Early identification and appropriate clinical intervention are of great significance to the reproductive endocrine function, bone health and long-term health such as terminal height of DP patients.

Intracranial germ cell tumors (iGCT) is a common cause of delayed puberty (DP).　A retrospective analysis was conducted on the clinical data of two patients with DP caused by iGCT. The disease characteristics of the two patients included DP, primary amenorrhea, growth retardation, short stature, subclinical hypothyroidism, etc. They were diagnosed as DP through imaging and related biochemical tests, and some sex hormone drugs were given on the basis of active treatment of the primary disease. Two patients had varying degrees of development in height, weight and secondary sexual characteristics, and further treatment condition is being tracking by us. Early identification and appropriate clinical intervention are of great significance to the reproductive endocrine function, bone health and long-term health such as terminal height of DP patients.

Objective To explore the effective chemical components of Fuzhenghuaxian decoction and its mechanism of action in the treatment of liver fibrosis based on UPLC-Q-TOF-MS combined with network pharmacology and experimental verification.Methods UPLC-Q-TOF-MS/MS technology was used to analyze the active components of Fuzhenghuaxian decoction.The potential targets and mechanisms of FZHXD were analyzed from the aspects of pharmacokinetics,target information,GO enrichment and KEGG Pathway.FZHXD was used to verify the key pathways and targets screened by network pharmacology in mice with liver fibrosis induced by carbon tetrachloride(CCl4),and further improve the accuracy of its prediction results.Results A total of 35 effective chemical components were identified,mainly saponins and organic acids.Through network pharmacology screening,669 drug targets,440 common targets,5 core targets and 11 key active ingredients were obtained.The main signal transduction pathways were predicted to be PI3K-Akt,FOXO,NF-κB signaling pathway,etc.In vivo experiments confirmed that FZHXD improved liver fibrosis in mice through PI3K/AKT,FOXO and NF-κB signaling pathways.Conclusion Fuzhenghuaxian decoction has the characteristics of multi-pathway and multi-target in anti-liver fibrosis by down-regulating PI3K,AKT,NF-κB and up-regulating FOXO.

Understanding the regulatory networks for germ cell fate specification is necessary to developing strategies for improving the efficiency of germ cell production in vitro. In this study, we developed a coupled screening strategy that took advantage of an arrayed bi-molecular fluorescence complementation (BiFC) platform for protein-protein interaction screens and epiblast-like cell (EpiLC)-induction assays using reporter mouse embryonic stem cells (mESCs). Investigation of candidate interaction partners of core human pluripotent factors OCT4, NANOG, KLF4 and SOX2 in EpiLC differentiation assays identified novel primordial germ cell (PGC)-inducing factors including BEN-domain (BEND/Bend) family members. Through RNA-seq, ChIP-seq, and ATAC-seq analyses, we showed that Bend5 worked together with Bend4 and helped mark chromatin boundaries to promote EpiLC induction in vitro. Our findings suggest that BEND/Bend proteins represent a new family of transcriptional modulators and chromatin boundary factors that participate in gene expression regulation during early germline development.
Animals ; Cell Differentiation/genetics* ; Chromatin/metabolism* ; Embryonic Stem Cells ; Germ Cells/metabolism* ; Germ Layers/metabolism* ; Mice

Objective To evaluate the application value of high-risk human papillomavirus (HR-HPV)DNA de-tection and ThinPrep liquid-based cytology testing(TCT)in cervical disease screening.Methods Cervical specimens of women with cervical lesions in a hospital between October 2012 and December 2013 were taken and performed hu-man papillomavirus DNA genotyping (HPV DNA)and ThinPrep liquid-based cytology testing(TCT).Positive pa-tients were performed colposcopy pathological detection.HPV DNA positive rates among different TCT groups, and different cervical lesion groups were compared,the sensitivity and specificity of TCT and HPV DNA detection, as well as differences between separate and joint detection were also compared.Results The positive rate of HPV DNA was 28.07% (1 045/3 723),most were HR-HPV (21 .57%,n=803),the major HR-HPV genotypes were HPV 16,58,52,and 18 type.HR-HPV positive rates were statistically different among different age groups(χ2 =31 .74,P <0.001 ),positive rate was highest in 20 -30 year old age group .Positive rate of TCT was 13.46%(n=501),a total of 971 patients were performed pathological detection,293 were positive.Patients were divided according to different TCT and different lesion type,χ2 testing of HR-HPV DNA positive rate showed that positive rate of HR-HPV had a increasing tendency with the increase in severity of diseases(all P <0.01).Pathological de-
tection was as a gold standard ,the sensitivity of HR-HPV DNA and TCT was 90.44%(265/293)and 85.32%(250/293)respectively,the sensitivity of joint HR-HPV DNA detection and TCT was 95.90%.In positive patho-logical group,the detection rate of TCT and HR-HPV DNA was 85.32% and 90.44%,respectively,joint detec-tion rate was 95.90%,the difference among three groups was significant (χ2 =18.185,P <0.001).Joint detection rate was higher than separate detection rate of TCT or HPV DNA.Conclusion HPV DNA detection is a useful supplement for cervical cancer screening,HPV DNA detection combined with TCT can reduce the misdiagnosis rate.

Objective:To establish cell lines stably expressing Rab5a and its the inactive mutant Rab5aN133I,analyze the effect of Rab5a on the expression of cytokines in LPS-stimulated RAW264.7 cells .Methods:RAW264.7 cells were transfected with Rab5a and its inactive mutant vector Rab5a N133I separately,and then screened by G418.Rab5a stable expressing cell lines were identified by Real time-PCR.The growth of the stable cell lines was analyzed by MTT assay.After the stable cell lines were stimulated by LPS for different time periods,the expression of iNOS,TNF-αand IL-6 was detected.Results:Rab5a and Rab5aN133I transfection resulted in elevated Rab5a mRNA expression compared with the control cells ( P<0.05 ).Rab5a overexpression enhanced the proliferation of RAW264.7 cells.However,the proliferation of Rab5aN133I cells was significantly slower than the control cells ( P<0.05).Overexpression of Rab5a promoted LPS-induced production of iNOS,TNF-αand IL-6 in RAW264.7 cells (P<0.01). Conversely,overexpression of Rab5aN133I abolished the stimulating effects of Rab5a.Conclusion: Rab5a promoted LPS-induced expression of iNOS,TNF-αand IL-6 in RAW264.7 macrophages in a GTP-binding ability-dependent manner.