As the important source of bone cells, osteoblasts are involved in bone formation and repair, and play a key role in maintaining bone balance. If the osteogenic differentiation process in vivo is disrupted, a variety of bone-related diseases may occur. Natural products, which have a wide range of sources, a wide variety of physiological activities, and few toxic side-effects, have been found in recent years to be able to regulate osteoblast differentiation. Based on the sources of natural products, this paper reviews the intervention of natural products from plant, animal and microbial sources on osteogenic differentiation, aiming to provide a theoretical basis for natural products in the treatment of bone diseases.

Andrographolide sulfonate (AS) is a sulfonated derivative of andrographolide extracted from Andrographis paniculata (Burm.f.) Nees, and has been approved for several decades in China. The present study aimed to investigate the novel therapeutic application and possible mechanisms of AS in the treatment of rheumatoid arthritis. Results indicated that administration of AS by injection or gavage significantly reduced the paw swelling, improved body weights, and attenuated pathological changes in joints of rats with adjuvant-induced arthritis. Additionally, the levels of tumor necrosis factor-alpha (TNF-α), interleukin-6 (IL-6), and IL-1β in the serum and ankle joints were reduced. Bioinformatics analysis, along with the spleen index and measurements of IL-17 and IL-10 levels, suggested a potential relationship between AS and Th17 cells under arthritic conditions. In vitro, AS was shown to block Th17 cell differentiation, as evidenced by the reduced percentages of CD4⁺ IL-17A⁺ T cells and decreased expression levels of RORγt, IL-17A, IL-17F, IL-21, and IL-22, without affecting the cell viability and apoptosis. This effect was attributed to the limited glycolysis, as indicated by metabolomics analysis, reduced glucose uptake, and pH measurements. Further investigation revealed that AS might bind to hexokinase2 (HK2) to down-regulate the protein levels of HK2 but not glyceraldehyde-3-phosphate dehydrogenase (GAPDH) or pyruvate kinase M2 (PKM2), and overexpression of HK2 reversed the inhibition of AS on Th17 cell differentiation. Furthermore, AS impaired the activation of phosphatidylinositol 3-kinase (PI3K)/protein kinase B (AKT) signals in vivo and in vitro, which was abolished by the addition of lactate. In conclusion, AS significantly improved adjuvant-induced arthritis (AIA) in rats by inhibiting glycolysis-mediated activation of PI3K/AKT to restrain Th17 cell differentiation.
Animals ; Th17 Cells/immunology* ; Diterpenes/pharmacology* ; Arthritis, Rheumatoid/metabolism* ; Proto-Oncogene Proteins c-akt/immunology* ; Glycolysis/drug effects* ; Cell Differentiation/drug effects* ; Phosphatidylinositol 3-Kinases/genetics* ; Rats ; Male ; Rats, Sprague-Dawley ; Humans ; Andrographis paniculata/chemistry* ; Arthritis, Experimental/drug therapy* ; Interleukin-17/immunology* ; Signal Transduction/drug effects*

Interactions between brain-resident and peripheral infiltrated immune cells are thought to contribute to neuroplasticity after cerebral ischemia. However, conventional bulk sequencing makes it challenging to depict this complex immune network. Using single-cell RNA sequencing, we mapped compositional and transcriptional features of peri-infarct immune cells. Microglia were the predominant cell type in the peri-infarct region, displaying a more diverse activation pattern than the typical pro- and anti-inflammatory state, with axon tract-associated microglia (ATMs) being associated with neuronal regeneration. Trajectory inference suggested that infiltrated monocyte-derived macrophages (MDMs) exhibited a gradual fate trajectory transition to activated MDMs. Inter-cellular crosstalk between MDMs and microglia orchestrated anti-inflammatory and repair-promoting microglia phenotypes and promoted post-stroke neurogenesis, with SOX2 and related Akt/CREB signaling as the underlying mechanisms. This description of the brain's immune landscape and its relationship with neurogenesis provides new insight into promoting neural repair by regulating neuroinflammatory responses.
Humans ; Ischemic Stroke ; Brain/metabolism* ; Macrophages ; Brain Ischemia/metabolism* ; Microglia/metabolism* ; Gene Expression Profiling ; Anti-Inflammatory Agents ; Neuronal Plasticity/physiology* ; Infarction/metabolism*

Objective:To investigate and distinguish the useful index of recognizing blood culture contamination and bloodstream infection when the blood cultures were positive in clinical practice, and to help clinicians to recognize blood culture contamination.Methods:A retrospective study was conducted to analyze the clinical data and distribution of bacteria of blood culture from 303 patients with positive blood culture from January to December 2021 in Huashan Hospital, Fudan University. Age, distribution of departments, positive time of blood culture, neutrophil ratio, procalcitonin and C-reactive protein (CRP) were compared between bloodstream infection group and blood culture contamination group. Mann-Whitney U test and chi-square test were used for statistical comparisons. The factors for recognizing blood culture contamination were analyzed by multivariate logistic regression. Results:A total of 303 patients with positive blood culture were enrolled and divided into two groups, including 237 cases in bloodstream infection group and 66 cases in blood culture contamination group, with the contamination rate of 21.78%. There was statistically significant difference in the distribution of departments between the two groups ( χ2=11.28, P=0.024). The proportion of age≥60 years old (52.7%(125/237) vs 36.4%(24/66)), the neutrophil ratio(0.86(0.79, 0.91) vs 0.82(0.75, 0.88)), the levels of procalcitonin (1.05(0.23, 6.64) μg/L vs 0.17(0.11, 0.72) μg/L) and CRP (60.55(24.83, 132.83) mg/L vs 40.64(8.98, 95.83) mg/L) in the bloodstream infection group were higher than those in blood culture contamination group, and the differences were statistically significant ( χ2=5.54, U=9 523.00, 10 906.00 and 9 278.50, respectively, all P<0.05). Among the 303 patients with positive blood culture, 253 patients had a single positive blood culture, 41 patients had twice positive blood cultures and nine patients had three times positive blood cultures. The highest contamination rate of detected pathogen was coagulase-negative staphylococcus (54.72%(29/53)) in the patients with single positive blood culture. The positive time of blood culture in the bloodstream infection group was 3(3, 4) d, which was shorter than 4(3, 4) d in the blood culture contamination group, and the difference was statistically significant ( U=6 521.00, P=0.026). Multivariate logistic regression analysis showed that when the time to positivity ≤3 days and procalcitonin level ≥2.00 μg/L, the positive blood culture results were more likely to be bloodstream infection (odds ratio ( OR)=2.16, 1.96, respectively, both P<0.05). Conclusions:Bacteria associated with blood culture, time to positivity and clinical index all play important roles in recognizing blood culture contamination. When the time to positivity ≤3 days and procalcitonin level ≥2.00 μg/L, the positive blood culture results are more likely to be bloodstream infection.

Objective:To explore the heterogeneity among keloids before and after radiotherapy and identify the changes of key cell subpopulations and their interactions utilizing single cell RNA sequencing technology.Methods:Four patients provided a total of 12 samples, each consisting of keloid tissue before and after radiotherapy and the normal skin tissue adjacent to the untreated keloid. The keloid was divided into left and right sides from the midline, and the left-side keloid was fractionally irradiated with 20 Gy electron beam in total in 4 consecutive days. The right-side keloid was irradiated with 10 Gy in 2 fractions before surgery and 10 Gy in 2 fractions after surgery.Results:A total of 25 573 fibroblasts were analyzed and categorized into nine subgroups (fibroblasts 1-9). The proportion of fibroblast-2 increased after radiotherapy ( t=4.70, P<0.05). The number of classical monocytes and macrophages increased after radiotherapy, but there was no significant difference due to the shorter time of sample taking at 2 d after radiotherapy ( P>0.05). Macrophages (4 723 cells) were further divided into four categories. CellPhoneDB analysis showed that type-3 macrophages interacted significantly more closely with fibroblasts than type-1 and type-2 macrophages. The most prominent signaling pathways for the interactions between type-3 macrophages and major fibroblast subtypes were the collagen signaling pathway and the chemerin signaling pathway. These interactions were more pronounced in the keloid samples after radiotherapy. Conclusions:The interactions between type-3 macrophages and fibroblasts (such as fibroblast-2) may serve as an important point for future studies on radio-sensitization of keloids.

Objective To investigate the effect and synaptic plasticity mechanism of dexmedetomi-dine(Dex)on alleviating cognitive impairment induced by myocardial ischemia/reperfusion(I/R)in aged mice.Methods Forty elderly male C57BL/6 mice were randomly divided into sham group,Sham+Dex group,I/R group,and I/R+Dex group,with 10 mice in each group.On the 7th day of reperfusion,cognitive function was evaluated using an eight arm water maze and novel ob-ject recognition.Electrophysiological recordings of hippocampal CA1 area was performed to meas-ure field excitatory postsynaptic potential(fEPSP)in in vivo long-term potentiation(LTP).The serum levels of cardiac troponin T(cTnT),lactate dehydrogenase(LDH),and N-terminal pro-B-type natriuretic peptide(NT-proBNP)were detected by ELISA.Western blotting was used to determine the expression of brain-derived neurotrophic factor(BDNF),tyrosine kinase receptor B(TrkB)-FL,and TrkB-T1 in hippocampal tissues.Golgi-Cox staining was utilized to observe the density of dendritic spines in hippocampal CA1 area.Immunofluorescence staining was performed to detect the expression of synaptic growth factor(SYN)and postsynaptic dense protein 95(PSD95).Results On the 7th day of reperfusion,the working memory error(WME),reference memory errors(RME)and total errors(TE)of the eight arm water maze and the expression lev-els of cTnT,LDH,NT-proBNP and TrkB-T1 were significantly higher,and the recognition index,fEPSP and expression levels of BDNF,TrkB-FL,SYN and PSD95 and density of dendritic spines were remarkably decreased in the I/R group and I/R+Dex group than the Sham group and Sham+Dex group(P＜0.05).The I/R+Dex group had obviously reduced TrkB-T1 expression(0.86±0.07 vs 1.51±0.18),and increased expression levels of BDNF(0.70±0.07 vs 0.39±0.06),TrkB-FL(0.67±0.10 vs 0.45±0.08),SYN(20.53±0.59 vs 16.50±1.05)and PSD95(17.73±0.52 vs 14.71±0.91)when compared with the I/R group(P＜0.05).Conclusion Dex exerts protective effect on cognitive impairment induced by myocardial I/R in elderly mice,and the specific mecha-nism may be related to alleviating cardiac injury,increasing the expression of BDNF,SYN and PSD95,improving the imbalance of TrkB-FL and TrkB-T1 ratio in hippocampal tissue,and enhan-cing synaptic plasticity.

Objective To explore the clinical efficacy of fluoxetine hydrochloride capsules and escitalopram oxalate tablets in the treatment of first-episode depression disorder.Methods Eighty patients with first-episode depression disorder who were hospitalized in the 14th district of Integrated Traditional Chinese and Western Medicine of Xiamen Xianyue Hospital from February to November 2023 were selected,and divided into fluoxetine hydrochloride capsules group(n=40)and escitalopram oxalate tablets group(n=40)according to random number method;Conducted the Hamilton depression scale(HAMD)estimates before treatment and four weeks after treatment,to analyze and compare the clinical efficacy of two groups.Results After four weeks of treatment,the HAMD scores of all patients significantly decreased compared to before treatment in this group(P＜0.01);There was no statistically significant difference between two groups(P＞0.05).There was no statistically significant difference in the total effective rate between two groups(P＞0.05).Conclusion Escitalopram oxalate tablets and fluoxetine hydrochloride capsules were both effective in treatment of the first-episode depression disorder patients,and the total effective rates of them were similar.

Objective:To summarize the morphological characteristics of diffuse large B-cell lymphoma (DLBCL) cells and investigate the prognosis value of the characteristics and the number of DLBCL cells in bone marrow.Methods:Retrospective study. We collected 79 cases newly diagnosed with DLBCL in the First Affiliated Hospital of Zhengzhou University from January 2020 to August 2022. 30 cases newly diagnosed without bone marrow involvement were selected as controls, whose mean age 58 years (30-82 years). The DLBCL cells were evaluated by the bone marrow smear, biopsy and flow cytometry separetely.The detection rate of DLBCL cells in the bone marrow was compared, to analyse the relationship between the morphological characteristics of DLBCL in the smear, clinical characteristics and flow cytometry parameters, and the prognostic value of DLBCL detected in the bone marrow smear and its quantity was analyzed. Logistic regression was used to analyze the correlation between the detection of DLBCL cells in bone marrow smears and the age, clinical stage, and the number of extraderules involved organs. Multivariate Cox regression was used to analyze the influence of DLBCL cells detection and its number on the prognosis of patients.Results:(1) The positive rates of DLBCL cells in bone marrow biopsy, bone marrow smear and flow cytometry were 4.86%, 5.14% and 9.27% respectively. (2) The morphological characteristics of 79 cases in bone marrow smears were described: more than 2 times the volume of the cell body of the lymphocyte, the shape was different, round or quasi-round or irregular shape, can be seen pseudopodia or protrusion; The volume of cytoplasm was moderate, vacuoles were visible, and a few perinuclear areas were visible. The nucleus were different in shape, round or quasi-round or irregularly shaped, with a majority of them having multiple nuclei and a few of them having delicate and loose chromatin. Most nucleoli were medium or large obviously, with a majority of them having 1-2 nucleoli and a few having more than 3.Sergiosomes and hemophagocytosis were observed in some DLBCL cases, tumor cell aggregation phenomenon was observed in a few DLBCL cases, occasionally pathological mitosis.(3) DLBCL cells in bone marrow smear was positively related to the age of patients, clinical stage and the number of extranodal organs involved(regression coefficient were 2.012, 2.754, 2.028, P<0.05);The volume of DLBCL cells in bone marrow smear was positively correlated with the ratio of CD4 and CD8(regression coefficient is 2.545, P<0.05);The vacuoles in cytoplasm and the pseudopod of tumor were both negative relationship with the quantity of CD38 expressed on DLBCL cells(regression coefficient was -2.465, -3.045, P<0.05); (4) DLBCL cells in bone marrow smear was an independent risk factor for PFS and OS( RR=7.059, P<0.05); RR=5.409, P<0.05). Conclusion:The appearace of DLBCL cells in bone marrow smear with prognosis, and could be used for clinical staging.

ObjectiveTo introduce the Hr gene of spontaneously mutated SHJH^hr mice into BALB/cAShjh inbred mice with clear genetic background,and provide a basis for study on the molecular mechanism of Hr gene mutation-induced abnormal phenotype and the application of this model.Methods Using a backcross-intercross breeding method guided by phenotypic monitoring, mutant genes from SHJH^hr mice bred by spontaneous mutation were introduced into inbred BALB/cAShjh mice by homozygous mutation introgression, and the mice were bred into BALB/cA.Cg.SHJH^hr (abbreviated as C.Cg.SHJH^hr) mice after 10 generations. The genotypes of 90 single nucleotide polymorphism (SNP) detection sites were analyzed in C.Cg.SHJH^hr mice by multiplex PCR library construction followed by next generation sequencing. Then 14 biochemical locus marker genes were detected in C.Cg.SHJH^hr mice according to the method of GB/T 14927.1-2008. Finally, whole genome exon sequencing was utilized to detect the mutated genes in this mouse. ResultsFrom May 2018 to March 2022, a total of 10 generations of backcross-intercross were conducted to complete the construction of the C.Cg.SHJH^hr mouse line. Among the 90 SNPs loci detected, except for rs13484115 and rs13484116, all the other loci had the same genotype as the recipient mice BALB/cAShjh. The results of biochemical marker gene detection showed that all the 14 loci of the mouse were the same as those of the recipient mouse. Whole genome exon sequencing found that the mouse had 109 site mutations compared with the recipient mouse strain, including 71 synonymous mutations, 1 stopgain, 37 missense mutations, and 20 genes involved in protein sequence alterations (including the reported Hr gene). ConclusionC.Cg.SHJH^hr mice were created. Through exon sequencing and genetic analysis, three Hr mutated genes and associated mutated genes that mainly cause phenotypic variations were identified, which provides a basis for expanding the application of C.Cg.SHJH^hr mice in biomedical research.

Objective To measure and analyze biological characteristics and aging phenotype of SHJH^hr mice and provide basic data for the application of the mouse model in aging mechanisms research and antiaging drug development. MethodsWith ICR mice of the same age as control group, the body mass growth data of SHJH^hr mice at the age of 3 to 16 weeks, the reproduction ability of 1 to 4 fetuses and the life cycle of SHJH^hr mice were measured. Blood routine (30 items) and serum biochemical indexes (25 items) of 6-week-old SHJH^hr mice were measured. The venous blood of 8-week-old SHJH^hr mice was collected for flow cytometry analysis to determine the content of immune cells. The aging bone structure of the cancellous bone and bone mineral density of SHJH^hr mice aged 4, 8 and 26 weeks were measured by micro-CT. Histopathological changes of bone and joint of 8-week-old mice were observed. ResultsCompared with ICR mice, the female and male body mass of SHJH^hr mice were significantly lower at the age of 16 weeks (P < 0.05), and the reproductive performance of female mice was low (P < 0.01) or did not have normal reproductive capacity. The shortest survival time of SHJH^hr mice was 57 weeks and the longest was 71 weeks, which was shorter than those of normal ICR mice, showing obvious rapid aging phenomenon. At the same time, some physiological and biochemical indexes of blood and pathological changes of bone and cartilage tissues also showed the accelerated aging and abnormality of animal physiological functions. ConclusionSHJH^hr mice have some biological characteristics of rapid aging as well as some physiological and pathological changes caused by aging.