Objective To explore the mechanism of the intervention effect of Qizhu Xiaozheng Fang(QZXZF),a representative prescription of Yiqi Jianpi Jiedu(replenishing qi,strengthening spleen,and removing toxicity),in treating HBx-mediated liver cancer stem cells(LCSCs).Methods The Kaplan-Meier survival analysis was used to investigate the effects of Yiqi Jianpi Jiedu on the prognosis of patients with HBV-HCC.The network pharmacology method was utilized to predict the targets and pathways of QZXZF in treating HBx-related LCSCs(HBx-LCSCs).HBx-LCSCs cells were screened by stably transfecting HBx and serum-free culture.The therapeutic effect of QZXZF on HBx-LCSCs was tested in vitro,and its effect on stemness markers and PI3K/Akt pathway was verified by qRT-PCR and Western blot.Results Yiqi Jianpi Jiedu could improve the overall survival time of HBV-HCC patients.Combined with the results of network pharmacology,the mechanism of action of QZXZF against HBx-LCSCs was explored from the PI3K/Akt pathway.Compared with the blank vector group,HBx can promote the expression of stemness markers in HBx-LCSCs cells;compared with the HBx-LCSCs control group,QZXZF could significantly inhibit the proliferation and colony formation of HBx-LCSCs cells(in a concentration-dependent manner),and could reduce the expression of stemness markers(EpCAM,NANOG,SOX2,and OCT4),phosphorylated PI3K(p-PI3K)and phosphorylated AKT(p-AKT)proteins.Conclusion QZXZF may regulate HBx-mediated LCSCs through PI3K/AKT pathways,providing a reference for the mechanism of TCM intervention in LCSCs.

Nonunion of osteoporotic vertebral fractures (OVF), predominantly affecting the elderly, can lead to intractable pain, vertebral collapse, progressive kyphotic deformity, and neurological impairment, significantly compromising patients′ quality of life. There exists considerable debate on diagnosis and management of OVF, encompassing key issues such as clinical diagnosis and staging criteria for nonunion, surgical indications and procedure selection, and postoperative rehabilitation planning. Currently, there lacks standardized clinical guideline and expert consensus on the diagnosis and management of OVF nonunion in China. To address this gap, Minimally Invasive Surgery Group of Chinese Orthopedic Association, Osteoporosis Committee of Chinese Association of Orthopedic Surgeons, Prevention and Rehabilitation Committee for Osteoporosis of Chinese Association of Rehabilitation Medicine and Minimally Invasive Orthopedic Surgery Branch of China Association for Geriatric Care jointly organized domestic experts in spinal surgery, endocrinology, and rehabilitation to formulate the Clinical guideline for the diagnosis and treatment for nonunion of osteoporotic vertebral fractures ( version 2025), based on existing literature and clinical experience and adhering to principles of scientific rigor and practicality. The guideline provided 13 evidence-based recommendations encompassing diagnosis and treatment of OVF nonunion, aiming to standardize its clinical management.

Objective To explore the mechanism of the intervention effect of Qizhu Xiaozheng Fang(QZXZF),a representative prescription of Yiqi Jianpi Jiedu(replenishing qi,strengthening spleen,and removing toxicity),in treating HBx-mediated liver cancer stem cells(LCSCs).Methods The Kaplan-Meier survival analysis was used to investigate the effects of Yiqi Jianpi Jiedu on the prognosis of patients with HBV-HCC.The network pharmacology method was utilized to predict the targets and pathways of QZXZF in treating HBx-related LCSCs(HBx-LCSCs).HBx-LCSCs cells were screened by stably transfecting HBx and serum-free culture.The therapeutic effect of QZXZF on HBx-LCSCs was tested in vitro,and its effect on stemness markers and PI3K/Akt pathway was verified by qRT-PCR and Western blot.Results Yiqi Jianpi Jiedu could improve the overall survival time of HBV-HCC patients.Combined with the results of network pharmacology,the mechanism of action of QZXZF against HBx-LCSCs was explored from the PI3K/Akt pathway.Compared with the blank vector group,HBx can promote the expression of stemness markers in HBx-LCSCs cells;compared with the HBx-LCSCs control group,QZXZF could significantly inhibit the proliferation and colony formation of HBx-LCSCs cells(in a concentration-dependent manner),and could reduce the expression of stemness markers(EpCAM,NANOG,SOX2,and OCT4),phosphorylated PI3K(p-PI3K)and phosphorylated AKT(p-AKT)proteins.Conclusion QZXZF may regulate HBx-mediated LCSCs through PI3K/AKT pathways,providing a reference for the mechanism of TCM intervention in LCSCs.

Nonunion of osteoporotic vertebral fractures (OVF), predominantly affecting the elderly, can lead to intractable pain, vertebral collapse, progressive kyphotic deformity, and neurological impairment, significantly compromising patients′ quality of life. There exists considerable debate on diagnosis and management of OVF, encompassing key issues such as clinical diagnosis and staging criteria for nonunion, surgical indications and procedure selection, and postoperative rehabilitation planning. Currently, there lacks standardized clinical guideline and expert consensus on the diagnosis and management of OVF nonunion in China. To address this gap, Minimally Invasive Surgery Group of Chinese Orthopedic Association, Osteoporosis Committee of Chinese Association of Orthopedic Surgeons, Prevention and Rehabilitation Committee for Osteoporosis of Chinese Association of Rehabilitation Medicine and Minimally Invasive Orthopedic Surgery Branch of China Association for Geriatric Care jointly organized domestic experts in spinal surgery, endocrinology, and rehabilitation to formulate the Clinical guideline for the diagnosis and treatment for nonunion of osteoporotic vertebral fractures ( version 2025), based on existing literature and clinical experience and adhering to principles of scientific rigor and practicality. The guideline provided 13 evidence-based recommendations encompassing diagnosis and treatment of OVF nonunion, aiming to standardize its clinical management.

Objective To investigate the effect of age on the incidence of cirrhosis and liver cancer in patients with chronic hepatitis B.Methods 279 patients with chronic hepatitis B were divided into the senior group and the younger group according to the age of the patients.The cumulative incidence of cirrhosis and liver cancer during 25 years of follow-up was calculated by using SPSS and R language through the long-term follow-up of HIS system,and the risk factors were analyzed by multivariate logistic regression.Results During follow-up,24 cases developed cirrhosis and 12 cases developed liver cancer.The cumulative incidence of liver cirrhosis was 1.5%,2.1%,5.4%,11.6%and 15.5%in the 5-year,10-year,15-year,20-year and 25-year group,and 5.5%,9.8%,22.9%,29.0%and 52.1%in the elderly,respectively.The difference between the younger age group and senior age group was statistically significant(P<0.001).A total of 2 risk factors(age and follow-up time)were included in the regression model.Two cases in the younger group developed into liver cancer after 17 and 21 years of follow-up,respectively.The cumulative incidence rates at 5,10,15,20 and 25 years were 1.8%,3.8%,18.5%,21.8%and 26.7%.A total of five factors(initial age,HBV-DNA load,HBV-DNA turned negative before the end-point,follow-up time,and sex)were included in the regression model.Conclusions The incidence of cirrhosis and liver cancer in CHB patients aged≥40 years,especially in male patients,is significantly higher than younger CHB patients.Timely initiation of antiviral therapy can delay disease progression and reduce the incidence of termi-nal liver disease.Whether antiviral therapy should be initiated for people aged 30 to 40 years remains to be studied.

Objective:To explore the effect and underlying mechanism of casein kinase 2 interacting protein-1 (CKIP-1) on hepatocyte apoptosis in nonalcoholic fatty liver disease (NAFLD).Methods:Experimental study. An NAFLD cell model was established by inducing human hepatoma cell line, HepG 2 cells, with oleic acid (OA). Flag-CKIP-1 expression vector and shRNA-CKIP-1 were transfected into HepG 2 cells. Flow cytometry was used to detect the effect of CKIP-1 on the activity and apoptosis of NAFLD hepatocytes. The levels of apoptosis-related proteins were detected by Western blot. CKIP-1 knockout mice in C57BL/6 back-ground were fed with either standard or high-fat diet for 8 weeks. Apoptosis-related signal proteins in NAFLD hepatocytes were detected by immunohistochemistry. Results:After CKIP-1 was transfected into HepG 2 cells, the degree of OA induced cell liposis was significantly reduced ( P<0.05). Annexin V-FITC/PI flow cytometry showed that CKIP-1 reduced the apoptosis of steatotic hepatocytes. Overexpression of CKIP-1 could significantly inhibit the expression of caspase-3 and caspase-9 and increase the expression of Bcl-2/Bax ( P<0.05). Knockdown of CKIP-1 could increase the expression of caspase-3 and caspase-9 ( P<0.05). CKIP-1 knockout could further increase the expression of caspase-3 and caspase-9 in NAFLD mice ( P<0.01, P<0.05), and further decrease the expression of Bcl-2/Bax ( P<0.05). Conclusion:CKIP-1 inhibited the apoptosis of steatotic hepatocytes by up-regulating the expression of apoptosis inhibitor gene, Bcl-2/Bax, and affecting the proteases, caspase-3 and caspase-9.

Fusobacterium nucleatum is an opportunistic pathogenic bacterium that can be enriched in colorectal cancer tissues, affecting multiple stages of colorectal cancer development. The two-component system plays an important role in the regulation and expression of genes related to pathogenic resistance and pathogenicity. In this paper, we focused on the CarRS two-component system of F. nucleatum, and the histidine kinase protein CarS was recombinantly expressed and characterized. Several online software such as SMART, CCTOP and AlphaFold2 were used to predict the secondary and tertiary structure of the CarS protein. The results showed that CarS is a membrane protein with two transmembrane helices and contains 9 α-helices and 12 β-folds. CarS protein is composed of two domains, one is the N-terminal transmembrane domain (amino acids 1-170), the other is the C-terminal intracellular domain. The latter is composed of a signal receiving domain (histidine kinases, adenylyl cyclases, methyl-accepting proteins, prokaryotic signaling proteins, HAMP), a phosphate receptor domain (histidine kinase domain, HisKA), and a histidine kinase catalytic domain (histidine kinase-like ATPase catalytic domain, HATPase_c). Since the full-length CarS protein could not be expressed in host cells, a fusion expression vector pET-28a(+)-MBP-TEV-CarS_cyto was constructed based on the characteristics of secondary and tertiary structures, and overexpressed in Escherichia coli BL21-Codonplus(DE3)RIL. CarS_cyto-MBP protein was purified by affinity chromatography, ion-exchange chromatography, and gel filtration chromatography with a final concentration of 20 mg/ml. CarS_cyto-MBP protein showed both protein kinase and phosphotransferase activities, and the MBP tag had no effect on the function of CarS_cyto protein. The above results provide a basis for in-depth analysis of the biological function of the CarRS two-component system in F. nucleatum.
Humans ; Histidine Kinase/metabolism* ; Fusobacterium nucleatum/metabolism* ; Automobiles ; Protein Kinases/genetics* ; Escherichia coli/metabolism* ; Colorectal Neoplasms

Based on polyketide syntheses gene (PKS) and non-ribosomal peptide synthetases gene (NRPS), one strain with high anti-pathogenic activity was screened from 77 strains isolated from Arctic marine sediments and identified. By optimizing the composition of culture medium and fermentation conditions, the production of this strain's active metabolites was improved and the main metabolites were identified by HRMS, ¹H NMR and ¹³C NMR. The antibacterial spectrum of the main metabolites and the effect of the metabolites on cucumber Fusarium wilt were also determined. The results showed that the strain was Bacillus velezensis and it showed growth promoting effect on plants. When the strain was cultured in 5 g/L maltose, 10 g/L tryptone, 10 g/L sodium chloride, at 30 ℃, 150 r/min for 60 h, the diameter of the inhibition zone increased from (16.23±0.42) to (24.42±0.57) mm. The metabolites of this strain mainly contain macrolide compound macrolactin A, which has antagonistic effect on a variety of pathogenic bacteria and fungi. Cucumber seedling experiments showed that the metabolites of this strain had a protective effect on cucumber Fusarium wilt, and showed a good potential for development and application as a biocontrol agent.
Polyketides/pharmacology* ; Fungi ; Bacteria ; Fusarium/genetics* ; Anti-Bacterial Agents/pharmacology* ; Peptide Synthases/genetics*

Cyclic lipopeptide has extensive application prospect in the field of medicine due to its unique chemical structure and biological activity. This study aims to obtain high purity of cyclic lipopeptide monomer from Bacillus amyloliquefaciems strain Q-426, and illuminate preliminary antitumor mechanism of C-15 Bacillomycin D and C-16 Bacillomycin D. Firstly, crude cyclic lipopeptide solution was prepared by two-steps purification of acid precipitation and double-resins chromatography. In order to obtain purer product preparative HPLC was utilized to separate and purify cyclic lipopeptide. Component 1 and component 2 were detected as C-15 Bacillomycin D and C-16 Bacillomycin D by HPLC-MS and ESI-MS/MS. Secondly, the effect of C-15 Bacillomycin D, C-16 Bacillomycin D and their mixture (1:1, mol:mol) on cell proliferation was measured using human cancer cells (Hela, MG, Hep-G2 and HT-29). The cyclic peptide showed a dose dependent manner on the cell proliferation inhibition of Hela and MG cells. Finally, the results of the scratch wound healing assay and FACS analysis revealed that C-16 Bacillomycin D can effectively influence the cells migration and the cells treated with C-16 Bacillomycin D showed typical apoptotic morphology with the increase of drug concentration in the early apoptosis, late apoptosis percentage increased, and G₀G₁ arrest was induced significantly.

Objective To analyze the status of sleep disorders in patients with functional gastrointestinal disease (FGID) and its relation with symptom characteristics .Methods From January to December 2014 ,questionnaire was carried out in FGID patients who met the Rome Ⅲ criteria and visited the outpatient department of gastroenterology at six third-level general hospitals in Tianjin City to assess the severity of symptoms ,sleep quality (Pittsburgh sleep quality index ,PSQI) ,and psychological state (anxiety and depression) .Chi-square test and Mann-Whitney rank sum test were performed for statistical analysis .Results Among 931 patients with FGID ,651 (69 .92％ ) patients had sleep disorders and 280 (30 .08％ ) patients had no sleep disorders .Among 828 patients with functional dyspepsia (FD) ,360 (43 .48％ ) patients had sleep disorders complicated with and depression .Among 292 patients with irritable bowel syndrome (IBS ) , 138 (47 .26％ ) had sleep disorders complicated with anxiety and depression .Among 618 patients with FD complicated with sleep disorders , 70 (11 .33％ ) patients overlapped with IBS ;among 210 patients with FD ,but without sleep disorder ,11 (5 .24％ ) patients overlapped with IBS and the percentage of the former was higher than the latter ,and the difference was statistically significant (χ2 =6 .580 , P=0 .01) .The proportion of lower abdominal pain ,sheep fecal or hard stool ,laborious defecation or incomplete defecation in FGID patients without sleep disorder were 22 .14％ (62/280) ,11 .79％ (33/280) ,19 .29％ (54/280) and 27 .86％ (78/280) ,respectively ;which were lower than those of FGID patients with sleep disorders (36 .10％ (235/651) ,21 .20％ (138/651) ,32 .41％(211/651) and 44 .39％ (289/651));and the differences were statistically significant (χ2 =17 .552 ,11 .569 , 16 .566 and 22 .419;all P<0 .01) .FGID patients with sleep disorders have more severe symptoms such as lower abdominal pain , lower abdominal discomfort (non-pain ) , sheep fecal or hard stool , laborious defecation incomplete defecation , and urgency than FGID patients without sleep disorders ;and the differences were statistically significant (Z= -4 .423 ,-1 .973 ,-3 .360 ,-4 .467 ,-4 .550 and -2 .420 ;all P<0 .05) . Conclusions Sleep disorders ,anxiety and depression often coexist in patients with FGID .Sleep disorders are closely related with lower gastrointestinal symptoms in patients with FGID .