Background and Objectives: Osteoblasts are derived from bone marrow mesenchymal stem cells (BMMSCs) and playimportant role in bone remodeling. While our previous studies have investigated the cell subtypes and heterogeneity in osteoblasts and BMMSCs separately, cell-to-cell communications between osteoblasts and BMMSCs in vivo in humans have not been characterized. The aim of this study was to investigate the cellular communication between human primary osteoblasts and bone marrow mesenchymal stem cells. Methods: and Results: To investigate the cell-to-cell communications between osteoblasts and BMMSCs and identifynew cell subtypes, we performed a systematic integration analysis with our single-cell RNA sequencing (scRNA-seq) transcriptomes data from BMMSCs and osteoblasts. We successfully identified a novel preosteoblasts subtype which highly expressed ATF3, CCL2, CXCL2 and IRF1. Biological functional annotations of the transcriptomes suggested that the novel preosteoblasts subtype may inhibit osteoblasts differentiation, maintain cells to a less differentiated status and recruit osteoclasts. Ligand-receptor interaction analysis showed strong interaction between mature osteoblasts and BMMSCs. Meanwhile, we found FZD1 was highly expressed in BMMSCs of osteogenic differentiation direction. WIF1 and SFRP4, which were highly expressed in mature osteoblasts were reported to inhibit osteogenic differentiation. We speculated that WIF1 and sFRP4 expressed in mature osteoblasts inhibited the binding of FZD1 to Wnt ligand in BMMSCs, thereby further inhibiting osteogenic differentiation of BMMSCs. Conclusions Our study provided a more systematic and comprehensive understanding of the heterogeneity of osteogenic cells. At the single cell level, this study provided insights into the cell-to-cell communications between BMMSCs and osteoblasts and mature osteoblasts may mediate negative feedback regulation of osteogenesis process.

Objective:The study aimed to investigate the effects of exercise on the expression of ferroptosis-related proteins and the associated mechanisms in a post-traumatic stress disorder(PTSD)model induced by transient foot shock.Methods:Thirty-five male C57BL/6J mice(8 weeks old)were randomly divided into three groups:Control group,PTSD group,and Exe+PTSD group.PTSD model mice were established by foot-shock and PTSD behaviors were assessed using a fear conditioning box.Iron distribution areas were detected by Fe3+staining.Western Blot analysis were performed to determine the expression level of ferroptosis-related proteins in the mice.Results:Compared to the control group,the freezing time and Fe3+distribution areas in the cortex and hippocampus were significantly increased in the mice with PTSD(P＜0.05).The levels of ferroportin1(FPN1),glutathione peroxidase 4(GPX4),and brain-derived neurotrophic factor(BDNF)proteins were significantly decreased(P＜0.05),while the expression of high mobility group box 1(HMGB1)protein was significantly elevated(P＜0.05).Exercise significantly reduced freezing time and Fe3+accumulation in the cortex and hippocampus(P＜0.05),and it also up-regulated the expression of FPN1,GPX4,and BDNF proteins(P＜0.05),while down-regulating HMGB1 expression compared to the PTSD group(P＜0.05).Conclusion:This study demonstrates that PTSD induces the ferroptosis-related pathway in mice,and exercise can inhibit iron accumulation and reverse the expression of ferroptosis-related proteins induced by PTSD,there-by alleviating PTSD-like behavior in mice.

OBJECTIVE: To investigate the role of tumor necrosis factor receptor-associated factor 6 (TRAF6) in regulating Bacillus Calmette-Guérin (BCG)-induced macrophage apoptosis. METHODS: The expression of TRAF6 in peripheral blood samples of 50 patients with active tuberculosis (TB) and 50 healthy individuals were detected using quantitative real-time PCR (qPCR). RAW264.7 macrophages were infected with BCG at different MOI and for different lengths of time, and the changes in expressions of Caspase 3 and TRAF6 were detected with Western blotting and qPCR. In a RAW264.7 cell model of BCG infection with TRAF6 knockdown established using RNA interference technique, the bacterial load was measured and cell apoptotic rate and mitochondrial membrane potential (MMP) were determined with flow cytometry. The expression levels of TRAF6, Caspase 3, PARP, BAX and Bcl-2 in the cells were detected using Western blotting, and the expressions of TRAF6 and Caspase 3 were also examined with immunofluorescence assay. RESULTS: The expression of TRAF6 was significantly upregulated in the peripheral blood of patients with active TB as compared with healthy subjects (P < 0.001). In RAW264.7 cells, BCG infection significantly increased the expressions of Caspase 3 and TRAF6, which were the highest in cells infected for 18 h and at the MOI of 15. TRAF6 knockdown caused a significant increase of bacterial load in BCG-infected macrophages (P=0.05), lowered the cell apoptotic rate (P < 0.001) and reduced the expressions of Caspase 3 (P=0.002) and PARP (P < 0.001). BCG-infected RAW264.7 cells showed a significantly increased MMP (P < 0.001), which was lowered by TRAF6 knockdown (P < 0.001); the cells with both TRAF6 knockdown and BCG infection showed a lowered BAX expression (P=0.005) and an increased expression of Bcl-2 (P=0.04). CONCLUSION TRAF6 promotes BCG-induced macrophage apoptosis by regulating the intrinsic apoptosis pathway.
Apoptosis ; BCG Vaccine ; Caspase 3/metabolism* ; Humans ; Intracellular Signaling Peptides and Proteins ; Macrophages ; Mycobacterium bovis/metabolism* ; Poly(ADP-ribose) Polymerase Inhibitors ; TNF Receptor-Associated Factor 6/metabolism* ; bcl-2-Associated X Protein/metabolism*

Objective:To evaluate the application effect of the nurse-led catheter extraction assessment model for children in PICU.Methods:From January to May 2020, 100 children with short-term catheter in PICU of Guangzhou Women and Children Medical Center were selected by convenient sampling method as the experimental group, the need for urethral catheter indwelling was assessed daily using an evidence-based assessment scale in PICU children, and the unnecessary indwelling catheters were removed timely, and 109 children with indwelling urethral catheters from August to December 2019 were collected as the control group, the catheter was removed by the nurse on medical advice, recorded and compared days of indwelling of catheters, the incidence of patients with catheter-associated urinary tract infection , resetting of catheters and the length of stay in ICU between the two groups.Results:The median and interquartile spacing of the days with indwelling catheter were 5.0 (6.0) days in the experimental group and 6.0 (6.0) days in the control group ( Z=-2.01, P<0.05) . In the experimental group, the incidence of catheter-associated urinary tract infection was 1.000 percent (1/100), and in the control group, the incidence of catheter-associated urinary tract infection was 1.835 percent (2/109); in the experimental group, 2 cases of urethral catheter were reset, and in the control group, 2 cases of urethral catheter were reset; the median and interquartile spacing of the length of stay in ICU was 6.5 (7.0) days in the experimental group and 7.0 (8.0) days in the control group. The differences of the above three indexes between the two groups were statistically significant ( χ2=0.26, 0.01, Z=-0.96, all P>0.05). Conclusions:The nurse-led catheter extraction assessment model for children can effectively shorten the catheter indwelling days for children in PICU, which has certain clinical practice significance for reducing the incidence of catheter-associated urinary tract infection.

OBJECTIVE: To examine whether the combination of Naoxintong Capsule with standard care could further reduce the recurrence of ischemic stroke without increasing the risk of severe bleeding. METHODS: A total of 23 Chinese medical centers participated in this trial. Adult patients with a history of ischemic stroke were randomly assigned in a 1:1 ratio using a block design to receive either Naoxintong Capsule (1.2 g orally, twice a day) or placebo in addition to standard care. The primary endpoint was recurrence of ischemic stroke within 2 years. Secondary outcomes included myocardial infarction, death due to recurrent ischemic stroke, and all-cause mortality. The safety of drugs was monitored. Results were analyzed using the intention-to-treat principle. RESULTS: A total of 2,200 patients were enrolled from March 2015 to March 2016, of whom 143 and 158 in the Naoxintong and placebo groups were lost to follow-up, respectively. Compared with the placebo group, the recurrence rate of ischemic stroke within 2 years was significantly lower in the Naoxintong group [6.5% vs. 9.5%, hazard ratio (HR): 0.665, 95% confidence interval (CI): 0.492-0.899, P=0.008]. The two groups showed no significant differences in the secondary outcomes and safety, including rates of severe hemorrhage, cerebral hemorrhage and subarachnoid hemorrhage (P>0.05). CONCLUSION The combination of Naoxintong Capsule with standard care reduced the 2-year stroke recurrence rate in patients with ischemic stroke without increasing the risk of severe hemorrhage in high-risk patients. (Trial registration No. NCT02334969).
Adult ; Humans ; Secondary Prevention/methods* ; Ischemic Stroke ; Stroke/prevention & control* ; Cerebral Hemorrhage/complications* ; Double-Blind Method ; Platelet Aggregation Inhibitors

Objective To investigate the relationship between body composition and serum lipids and uric acid among adults in Maonan, and to analyze the effect of body composition changes on blood lipid and uric acid. Methods Totally 584 Maonan adult volunteers in Maonan village of Maonan Autonomous County in Guangxi, the age from 20 to 80 were recruited. The height was measured by the personal height tester; the body composition was measured by the ANITAMC-180 instrument; and the blood lipids and blood uric acid were measured by the Hitachi 7600 instrument. The obtained data were statistically analyzed by SPSS 20. 0. Results The age,height, weight, free fat mass, muscle mass, presumptive bone mass, body water, proptein,extracellular water, intracellular water, and waist-to-hip ratio were greater in Maonan men than in women (P<0. 05). However, whereas male fat content, body fat rate, and subcutaneous fat content were smaller than those of female (P < 0. 01). The total prevalence of hyperuricemia and hyperlipidemia in Maonan nationality was 13. 9% and 28. 4%, respectively. The prevalence of hyperuricemia and hyperlipidemia in males was higher than in females. In males, the body mass, body mass index, free fat mass, fat mass, muscle mass, presumptive bone mass, protein, extracellular water, body fat rate, visceral fat content, subcutaneous fat content and waist-hip ratio of the hyperlipidemia group were higher than the normal group (P<0. 05); and in females, the age, body mass index, fat mass, body fat rate, visceral fat content and waist-hip ratio of the hyperlipidemia group were higher than the normal group. In male, The body mass, free fat mass, presumptive bone mass, body water, extracellular water of the hyperuricemia group were higher than the normal group (P<0. 05); In females, the age, body mass, body mass index, fat mass, extracellular water, body fat ratio, muscle mass, visceral fat content, subcutaneous fat content, and waist-hip ratio of the hyperuricemia group were higher than the normal group. Conclusion The detection rate of hyperlipidemia and hyperuricemia in males of Guangxi Maonan nationality is all higher than that in females. The body composition is significant differences between the normal adults and the patients with hyperlipidemia and hyperuricemia of Maonan nationality in Guangxi.

Based on the data mining technology, the rules of acupoint selection and prescription were analyzed for impotence treated with acupuncture and moxibustion in ancient recorded in
Acupuncture Points ; Acupuncture Therapy ; Data Mining ; Erectile Dysfunction ; Humans ; Male ; Meridians ; Moxibustion ; Technology

Intestinal microecology is an important defense system in the human body. The intestinal flora is the core micro-ecosystem in the human intestine. It has a symbiotic relationship with the overall functions of the body. It has strong metabolic activity to maintain the normal functioning of the body and resist the invasion of various viral antigens in the body. Playing a protective function,the imbalanced intestinal microecology can cause various diseases. Polysaccharides can be extracted from a wide range of sources and have low toxicity and side effects. They have attracted wide attention because of their anti-tumor, anti-oxidant, anti-inflammatory and other biological activities. Studies have demonstrated that polysaccharides can regulate intestinal microecological disorders. According to the studies in recent years, this review summarizes that polysaccharides mainly modulate intestinal microecological disorders through regulating the composition of intestinal flora, improving the metabolism of the flora, and repairing the intestinal tract barrier. On the basis of these mechanisms of action, this paper elaborates the anti-tumor, immunomodulatory, and anti-inflammatory activities of polysaccharides. This paper can provide reference for the future research on the intestinal microecology-regulating mechanism and biological activities of polysaccharides.
Anti-Inflammatory Agents ; Ecosystem ; Gastrointestinal Microbiome ; Humans ; Intestines ; Polysaccharides/pharmacology*

Objective: Coronavirus Disease 2019 (COVID-19) has been diagnosed as an epidemic disease characterized by cold and dampness pathogens in TCM clinic. Due to many Chinese medicines with different functions were used in the treatment of COVID-19, it is very important to find the law of application of damp-removing traditional Chinese medicine with high frequency application, with view to providing a reference for the use and research of Chinese medicine to further control the pandemic. Methods: The publicly released diagnosis and treatment programs issued by the National Health Commission and Health Commission of provinces, autonomous regions and municipalities, and Chinese herbs prescription information in these were collected, a database was established, and Excel and Graphpad 8.0 software were used to analyze the frequency of use of various Chinese medicines, the frequency and property characters including five flavors (bitter, pungent, sweet, sour, and salty) and four natures (warm, hot, cool, and cold) and channel tropisms of Chinese medicine for removing dampness. Results: A total of 137 prescriptions of Chinese medicine for treating COVID-19 were collected, including 178 TCMs showing functions of resolving phlegm, relieving cough and asthma, resolving dampness, clearing damp and inducing dieresis, clearing heat, tonifying deficiency, and relieving exterior syndrome, in which the TCMs with the first four functions that we called the dampness-removing TCMs, accounted for 35.78%. Also a number of TCMs in the rest functions showed removing-dampness. The first four functions were divided into subfunctions including aromatic resolving dampness, clearing heat and drying dampness, drying dampness and tonifying spleen qi, drying dampness and removing phlegm, inducing diuresis and relieving swelling, inducing diuresis and relieving exterior syndrome. Among them, the most frequently used TCMs was Ephedrae Herba, followed by Citri Reticulatae Pericarpium, Pogostemonis Herba, Pinelliae Rhizoma, Poria, Scutellariae Radix, and Atractylodis Rhizoma. The property character analysis in the dampness-removing TCMs showed that bitter and pungent were largely present and sour and astringent were absent, warm and hot were dominant; And the lung, spleen, stomach, large intestine, bladder were main channel tropisms. Conclusion: Dampness-removing TCMs are the first important type of traditional Chinese medicine to be considered in the treatment of COVID-19 in Chinese medicine. The application of dampness-removing TCMs in the treatment of COVID-19 needs to be combined with its application law. This study may provide meaningful and useful information on further research to investigate the effective compounds from the dampness-removing Chinese medicine with high frequency application, and also provide a reference for the clinical treatment of COVID-19 accurately against dampness evil with dampness-removing traditional Chinese medicines.

Objective:To observe the effects of Analgecine (AGC) on middle cerebral artery ischemia-reperfusion injury in rats and its mechanism. Methods:A total of 61 Sprague-Dawley rats were divided into sham group (n = 11), sham-AGC group (n = 11), model group (n = 20) and model-AGC group (n = 19). The model group and the model-AGC group were occluded the middle cerebral arteries for 1.5 hours and reperfused (2 rats in each group unsuccessful). The sham-AGC group and the model-AGC group were injected AGC 20 U/kg through tail-vein, while the sham group and the model group were injected saline of same volume. Four rats in each group were tested heat shock proteins 70 (HSP70), Bcl-2 and Bax in brain with Western blotting 48 hours after injection. The other rats were assessed with Prehensile Traction Test seven days after injection, and then, four of each group were detected ionized calcium binding adapter molecule 1 (Iba1) and glial fibrillary acidic protein (GFAP) expression with immunohistochemistry. Results:The prehensile time increased in the model-AGC group compared with that of the model group (P < 0.01), with the increase of HSP70 and Bcl-2 (P < 0.01) and decrease of Iba1 and GFAP expression (P < 0.05). Conclusion:AGC may promote the recovery of motor function in rats with cerebral ischemia-reperfusion injury, which may associate with inhibiting cell apoptosis and neruoinflammatory response.