BACKGROUND: The addition of growth factiors is commonly applied to improve the osteogenic differentiation of stem cells. However, for human pluripotent stem cells (hPSCs), their complex differentiation processes result in the unknown effect at different stages. In this study, we focused on the widely used bone forming peptide-1 (BFP-1) and investigated the effect and mechanisms of its addition on the osteogenic induction of hPSCs as a function of the supplementation period. METHODS: Monolayer-cultured hPSCs were cultured in osteogenic induction medium for 28 days, and the effect of BFP-1 peptide addition at varying weeks was examined. After differentiation for varying days (0, 7, 14, 21 and 28), the differentiation efficiency was determined by RT–PCR, flow cytometry, immunofluorescence, and alizarin red staining assays. Moreover, the expression of marker genes related to germ layers and epithelial-mesenchymal transition (EMT) was investigated at day 7. RESULTS: Peptide treatment during the first week promoted the generation of mesoderm cells and mesenchymal-like cells from hiPSCs. Then, the upregulated expression of osteogenesis marker genes/proteins was detected in both hESCs and hiPSCs during subsequent inductions with BFP-1 peptide treatment. Fortunately, further experimental design confirmed that treating the BFP-1 peptide during 7–21 days showed even better performance for hESCs but was ineffective for hiPSCs. CONCLUSION The differentiation efficiency of cells could be improved by determining the optimal treatment period.Our study has great value in maximizing the differentiation of hPSCs by adding osteogenesis peptides based on the revealed mechanisms and promoting the application of hPSCs in bone tissue regeneration.

OBJECTIVE: This study aimed to explore the association between humidity exposure and the risk of cardiovascular disease (CVD), utilizing follow-up data and relative humidity (RH) metric assessments. METHODS: We extracted the baseline data from the China Hypertension Survey (CHS) of 24,510 enrolled participants aged ≥ 35 years without a history of CVD between 2012 and 2015 and followed them up from 2018 to 2019. The National Meteorological Information Center (NMIC) of the China Meteorological Administration (CMA) provided the quality-controlled relative humidity (RH) datasets. Cox proportional hazards models were used to estimate hazard ratios ( HRs) for CVD in relation to RH. RESULTS: During the follow-up period (2018-2019), 973 patients with CVD were identified. The HR of CVD risk was 1.17 (95% CI: 1.04-1.31) per 10% increase in summer mean RH. Compared with participants in the 3 ^rd quintile group, those in the 1 ^st and 5 ^th quintiles of RH had a higher risk of CVD. For summer mean RH, the HRs (95% CIs) for the 1 ^st and 5 ^th quintiles were 1.34 (1.04-1.71) and 1.44 (1.14-1.83), respectively. The relationship ("U" shape) between summer mean RH and the risk of CVD was nonlinear. Stratified analyses indicated that the risk of CVD was substantially influenced by the summer mean RH in female, older individuals, and those in southern China. CONCLUSION Unsuitable (too high or low) humidity environments affect the risk of CVD. Our study highlights those future policies for adapting to climate change should consider the humidity-CVD relationship.
Humans ; Humidity/adverse effects* ; Cardiovascular Diseases/etiology* ; Female ; Male ; Middle Aged ; Prospective Studies ; China/epidemiology* ; Adult ; Aged ; Risk Factors ; Proportional Hazards Models ; Seasons

In recent years ，biomimetic nanodelivery system based on cell membrane coating has developed rapidly and shows better biocompatibility and efficacy than traditional nanodelivery systems in a variety of diseases . Macrophages，as members of the immune system ，are closely related to the occurrence and development of a variety of diseases . Macrophages are derived from monocytes and can be polarized into M 1 and M 2 types after corresponding stimulation ： M1 macrophages involved in the proinflammatory reaction and M 2 macrophages involved in the inflammatory reaction . This paper reviews the application status of biomimetic nanoparticles coated with macrophage membrane in disease targeted therapy in recent years . Biomimetic nanoparticles coated with macrophage membrane has shown its high targeting and low immunogenicity in the treatment of malignant tumors （breast cancer ，colorectal cancer ，melanoma，glioma），Alzheimer’s disease ，liver ischemia -reperfusion injury ，atherosclerosis and so on . However，the research of Biomimetic nanoparticles coated with macrophage membrane currently focuses on anti -tumor research and is still in the laboratory research stage .

【Objective】 To assess the prevalence and treatment of high cardiovascular disease （CVD） risk， and identify individual characteristics related to high CVD risk. 【Methods】 Based on the data of the China Patient-Centered Evaluative Assessment of Cardiac Events （PEACE） Million Persons Project （MPP） from 2016 to 2019， this study enrolled local residents aged 35 to 75 years from 39 counties or districts in Northwest China. Rates of high CVD risk and individual characteristics were assessed in the overall study population. Statin and aspirin use was also evaluated among those at high risk for CVD. Multivariable mixed models were fitted to evaluate the relationship between individual characteristics and high CVD risk. 【Results】 Among 364 537 participants， the average age was （54.6±9.7）years， and 5.8% was at a high risk for CVD. Multivariate mixed models showed that individuals who were currently using alcohol， overweight or obese tended to have a high risk for CVD， while married persons， those with a higher education level or a higher household income were correlated with a lower risk for CVD （all P<0.05）. Among high-risk persons， hypertension was the most prevalent risk factor （98.1%）， and only 1.3% and 3.5% reported their use of statins and aspirin， respectively. 【Conclusion】 Of the 364 537 participants， about 1 in 17 had a high risk for CVD. Among those at a high CVD risk， only less than 4% reported taking statins or aspirin. These findings indicate that there is still much room for risk mitigation in this population in China.

Objective:To investigate the effect of neutrophils on T lymphocyte function in septic mice and the role of CD80/cytotoxic T lymphocyte antigen-4 (CTLA-4) signaling pathway in this modulated effects.Methods:① In vivo experiment: 6-8 weeks old male C57BL/6 mice were divided into sham operation group (Sham group, n = 20), Sham+CTLA-4 antibody treatment group (Sham+aCTLA-4 group, n = 20), cecal ligation and perforation (CLP) induced sepsis model group (CLP group, n = 30) and CLP+CTLA-4 antibody treatment group (CLP+aCTLA-4 group, n = 30) according to the random number table. CLP was used to reproduce mouse sepsis model. The mice in the Sham group were treated identically but their cecums were neither punctured nor ligated. In CTLA-4 antibody treatment groups, 50 μg CTLA-4 antibody was injected intraperitoneally 6 hours and 24 hours after the operation. Forty-eight hours after operation, 6 mice in Sham group and Sham+aCTLA-4 group, 14 mice in CLP group and CLP+aCTLA-4 group were randomly selected to detect the expression of CD69 in spleen. At the same time, spleen, bone marrow and peripheral blood were collected, and the expression of CD80 on neutrophils was detected by flow cytometry. The expression of CTLA-4 on the surface of T lymphocytes in spleen was detected by immunofluorescence and flow cytometry. The remaining mice in each group were used to observe the 96-hour survival after operation.② In vitro experiment 1: neutrophils were extracted from bone marrow of healthy mice and stimulated with LPS (1 mg/L) for 4, 8 and 12 hours respectively. The control group was added with the same amount of phosphate buffered saline (PBS) at each time point, and the expression of CD80 was detected at each time point.③ In vitro experiment 2: splenic T lymphocytes of healthy mice were extracted and divided into PBS control group, LPS group (final concentration of LPS 1 mg/L), neutrophil group and neutrophil+LPS group. In the latter two groups, the co-culture model of neutrophils and T lymphocytes was established, and then the corresponding treatment was given to detect the expression of CTLA-4 on the surface of T lymphocytes. With the above four groups as controls, CTLA-4 antibody treatment groups (final concentration of CTLA-4 antibody 50 mg/L) were set up respectively. After 48 hours, the level of interleukin-2 (IL-2) in the cell supernatant was detected by enzyme linked immunosorbent assay (ELISA). Results:① Results of in vivo experiment: compared with Sham group, the expression of CD80 on neutrophils in spleen, bone marrow and peripheral blood was significantly up-regulated, while the expression of CTLA-4 on the surface of T lymphocytes was significantly increased [(9.98±0.84)% vs. (3.48±0.64)%, P < 0.05]. It suggested that neutrophils may affect T lymphocytes function through CD80/CTLA-4 pathway in sepsis. Compared with CLP group, CTLA-4 antibody could significantly improve the 96-hour cumulative survival rate of CLP mice (56.25% vs. 18.75%, P < 0.05), and increase the expression of CD69 on the surface of T lymphocytes. It suggested that CTLA-4 antibodies might increase T lymphocytes activation in sepsis and improve survival. ② Results of in vitro experiment: with the prolongation of LPS stimulation, the expression of CD80 on neutrophils gradually increased in time-dependent manner as compared with PBS control group [4 hours: (6.35±0.40)% vs. (3.41±0.40)%, 8 hours: (8.57±0.64)% vs. (3.09±0.27)%, 12 hours: (19.83±1.06)% vs. (5.16±0.36)%, all P < 0.05]. Compared with PBS control group, the expression of CTLA-4 on CD4 +/CD8 + T lymphocytes was not significantly affected by LPS stimulation alone, but CTLA-4 was increased after co-culture with neutrophils [CD4 +: (4.92±0.30)% vs. (3.33±0.25)%, CD8 +: (4.26±0.21)% vs. (2.53±0.66)%, both P < 0.05], and the increased trend of CTLA-4 was more obvious after co-culture with LPS-stimulated neutrophils [CD4 +: (6.34±0.50)% vs. (3.33±0.25)%, CD8 +: (6.21±0.41)% vs. (2.53±0.66)%, both P < 0.05]. In the PBS control group and LPS group, CTLA-4 antibody had no significant effect on IL-2 secretion of T lymphocytes. Compared with PBS control group, co-culture with neutrophils could inhibit the secretion of IL-2 by T lymphocytes (ng/L: 1 938.00±68.45 vs. 2 547.00±218.00, P < 0.05), and the inhibitory effect of neutrophils stimulated by LPS was more obvious (ng/L: 1 073.00±34.39 vs. 2 547.00±218.00, P < 0.05). CTLA-4 antibodies could partially restore IL-2 secretion. In conclusion, after promoting the expression of CTLA-4 on the surface of T lymphocytes, neutrophils might mediate the inhibition of T lymphocytes function by reducing the production of IL-2. Conclusions:Neutrophils mediate T lymphocytes dysfunction in sepsis, and the CD80/CTLA-4 pathway plays an important role. The CTLA-4 antibody improves survival and T lymphocytes function in sepsis mice, which may be a new method of immunotherapy for sepsis.

Objective:To investigate the efficacy and safety of internet-based cognitive behavioral therapy for insomnia(iCBT-I) combined with estazolam for patients with chronic insomnia.Methods:Patients with chronic insomnia were randomly assigned to treatment group which were intervened with iCBFI combined with estazolam( n=46) and control group which were intervened with estazolam ( n=43) for 8 weeks according to random number table.Pittsburgh sleep quality index (PSQI) and state-trait anxiety inventory (STAI) were used to measure the anxious state, anxious trait and sleep quality at three time points: before intervention(T1), after one month(T2) and two months(T3). Treatment emergent symptom scale (TESS), blood routine, urine routine, liver and renal function and electrocardiogram were used to measure the safety.The dosage of estazolam was compared between the two groups after two months.χ 2 test and repeated measurement analysis of variance were performed by SPSS 19.0. Results:The PSQI scores of control group and treatment group were (10.41±2.48) vs (9.98±2.96) at T2 and (9.97±2.13) vs (7.82±1.57) at T3.The state anxiety scores of control group and treatment group were (57.27±2.74) vs (56.27±2.89) at T2 and (45.67±2.62) vs (42.67±2.97) at T3.The data of T2 and T3 were statistically significant compared with those before intervention(all P<0.05). Compared with the control group, the treatment group was better on treatment efficiency(86.96% vs 69.77%) at T3( P<0.05). PSQI score, subjective sleep quality, sleep efficiency, sleep disorders, sleep drugs, daytime dysfunction, drug maintenance and adverse reaction were significantly different between the two groups at T3 ( P<0.05). Conclusions:Internet-based cognitive behavioral therapy combined estazolam for insomnia can improve sleep quality, anxious state and trait for chronic insomnia patients.Good safety was improved, as well as reducing the need of drug.So it's worthy of clinic application.

Objective:To investigate the effect of heparin-binding protein (HBP) on the damage of vascular permeability in early burn.Methods:① Clinical research: 12 patients with severe burns admitted to Suzhou Hospital of Nanjing Medical University from January 1st to August 30th in 2019 were enrolled. Eight patients with severe trauma admitted to the same hospital during the same period were also enrolled as controls to explain the specificity of burn injury. Whole blood samples were obtained within 0.5 hour after admission. The white blood cell count (WBC), absolute value and ratio of neutrophils, and serum HBP levels were measured. Serum samples of 12 patients with severe burn were collected within 9 days after admission, and enzyme-linked immunosorbent assay (ELISA) was used to detect the levels of metabolism products of glycocalyx including syndecan-1 and hyaluronic acid (HA). The correlation between HBP and neutrophils ratio, syndecan-1 and HA were analyzed by linear correlation. ② Basic research: a 30% total body surface area (TBSA) Ⅲ° burn model of Sprague-Dawley (SD) rat aged 6-8 weeks was prepared. In low molecular weight heparin (LMWH) intervention group ( n = 5), 200 U/kg LMWH was injected subcutaneously immediately and every 2 hours after injury for 4 times in total; the burn group ( n = 5) was given the same amount of normal saline. No intervention was given to the normal control group ( n = 5). The peripheral venous blood was collected at 0, 2, 4, and 8 hours after injury, and the serum levels of HBP, syndecan-1 and HA were measured; the injury of glycocalyx on pulmonary vascular endothelial cells was observed under transmission electron microscope. Results:① Clinical research results: the WBC, neutrophils absolute value and ratio, and HBP levels were increased in 12 patients with severe burn and 8 patients with severe trauma. There was no significant difference in the WBC, absolute value and ratio of neutrophils between severe burn and severe trauma patients [WBC (×10 9/L): 14.5±6.1 vs. 10.8±3.6, the absolute value of neutrophils (×10 9/L): 12.0±5.9 vs. 9.0±4.0, the ratio of neutrophils: 0.81±0.10 vs. 0.79±0.14, all P > 0.05], but the HBP levels in the burn patients were significantly higher than those in the trauma patients (μg/L: 192.92±61.73 vs. 51.17±23.05, P < 0.01). Twelve patients with severe burns had a sharp increase in serum syndecan-1 and HA levels after burns, which continued to maintain high levels and peaked at the 9th day [syndecan-1 (μg/L): 16.02±0.39, HA (μg/L): 106.83±4.90]. The analysis showed that HBP was positively correlated with neutrophils ratio, syndecan-1 and HA in severe burn patients at the 1st day after admission ( r values were 0.805, 0.732 and 0.900, respectively, all P < 0.01). It indicated that the sharp increase of neutrophils after the burn released a lot of HBP, and the glycocalyx of the vascular endothelium was severely damaged. ② Basic research results: the levels of serum HBP, syndecan-1 and HA in the burn group were increased sharply as compared with the normal control group, and continued to increase with time, reaching a peak at 8 hours after burn. In the LMWH intervention group, the serum levels of HBP, syndecan-1 and HA were significantly lower than those in the burn group, and the difference was still statistically significant after 8 hours [HBP (μg/L): 6.47±0.25 vs. 12.48±0.08, syndecan-1 (μg/L): 19.06±1.48 vs. 25.92±3.34, HA (μg/L): 35.76±2.10 vs. 54.91±2.64, all P < 0.01]. The results of transmission electron microscopy showed that in the normal control group, the glycocalyx pulmonary vascular endothelial cells was continuous, evenly distributed and dense. The glycocalyx on pulmonary vascular endothelial cells of rats were significantly damaged and shed 2 hours after burn in the burn group, and no glycocalyx was observed at 8 hours. In the LMWH intervention group, the glycocalyx on pulmonary vascular endothelial cells was damaged and the phenomenon of shedding was significantly relieved, and the glycocalyx could be observed 8 hours after the injury. Conclusion:The massive exudation of body fluids and the significant increase of vascular permeability in patients in early burns may be related to the destruction of the glycocalyx on endothelial cells by HBP released from increased neutrophils.

Objective To compare the efficacy and complications of the extend endoscopic endonasal approach (EEA) and open transcranial approach for resection of craniopharyngiomas. Methods The clinical data from 46 patients with craniopharyngiomas with extend EEA and 54 patients with transcranial route in our department was analyzed retrospectively. The gross total resection (GTR) rate,length of hospital stays and complications of the two groups were compared. Results The tumor diameters of were larger in the endoscopic group than in the transcranial group (3.5 ± 1.3cm vs. 3.0±0.8 cm, P<0.05). The endoscopic group had a greater GTR rate (67.4％vs. 46.3％, P<0.05)and improved visual outcome(84.2％ vs. 59.5％,P<0.05),but lower rate of hypopituitarism (56.5％ vs. 75.9％,P<0.05)and permanent diabetes insipidus (51.4％ vs.72.7％,P<0.05). On the contrast, the endoscopic group had a greater rate of cerebrospinal fluid leak(4.3％ vs. 0.0％,P>0.05)and longer hospital stays(17.0±3.6 d vs. 13.1±2.3 d,P<0.01). Hyposmia(34.8％)and hemorrhinia (2.2％)only happened in the endoscopic group. Conclusion Compared with transcranial route, the extend EEA for craniopharyngiomas is minimal invasion and effective, which can effectively improve the GTR rate and reduce the clinical symptoms.

Objective To investigate the value of MSCT in evaluating pulmonary functional changes in silicosis.Methods 56 cases of silicosis and 10 healthy people(as control group) underwent inspiratory and expiratory MSCT scans and pulmonary functional test one week later.The CT findings including silicotic nodules,large opacity,emphysema,reticular opacities,bronchiectasis and air trapping were recorded and graded subjectively on CT images.Air retention and emphysema were quantified using the software of Pulmo.CT scores were correlated with spirometric findings by using spearman rank correlative analysis in comparison with pulmonary functional index.Results The scores of CT features except for reticular shadows were of significant difference between silicotic group and control group,but not between simple silicotic group and complex silicotic group.The scores of air retention and emphysema on CT were significantly negative correlation.The reticular shadows were of positive correlation with FEF 75%,FEF 50% and FEV1,but on correlation with FEV1/FVC.There were no correlation between the scores of silicotic nodules,bronchiectasis,large shadows and pulmonary functional test.There was obvious correlation between air retention,the scores of emphysema on CT and the index of pulmonary function in obstruction of small airway.Conclusion MSCT is of important value in evaluating the damage of pulmonary function in silicosis.