Aim To observe the inhibitory effect of Alpha-momorcharin (α-MMC) on the inflammatory cytokine storm of Ml-type inflammatory macrophages induced by LPS and explore its possible targeting mechanism. Methods Western blot was used to detect the expression of WIL2-S B lymphocytes, H9 T lymphocytes, THP-1 monocytes and M0 macrophages LRP1 receptor protein. CCK-8 method was used to detect the survival rate of the four cells. ELISA was used to detect the expression level of inflammatory cytokines in Ml macrophages. Western blot was used to detect the expression of TLR4 signaling pathway-related protein in Ml macrophages. Results Macrophages had a high density of LRP1 receptors consistent with monocytes; the survival rate of α-MMC on the four cells was positively correlated with the density of this receptor; α-MMC inhibited the expression of inflammatory cytokinesTNF-α, IL-lβ, IL-6, IL-8, MlP-lα and MCP-1 in Ml macrophages in a dose-and time-dependent manner; α-MMC showed significant inhibition to TAKl/pTAK1, p-JNK, p-APl and p-p65 signaling proteins of the TLR4 signaling pathway, and this inhibition could be blocked by the LRP1 receptor blocker RAP. Conclusions α-MMC selectively inhibits macrophage inflammatory cytokine synthesis by inhibiting TAK1 of the TLR4 signaling pathway, which in turn inhibits the downstream NF-ΚB and MAPK pathways, mediated by the LRP1 receptor. The selective immunosuppressive effect of α-MMC on macrophages may make it a very promising agent for the treatment of acute infectious macrophage activation syndrome (MAS).

[Abstract] Objective To investigate the branching pattern of the ureteric bud and the number of the nephron induced by each ureteric bud tip, through the three-dimensional tracing of the ureteric tree, combined with the morphological analysis and measurement of the ureteric tree. Methods The kidneys were obtained from three mice at various developing time points and prepared for paraffin and epoxy sections. Then the microscopic images were digitized and aligned from these sections. Based on the computer-assisted tracing and visualization of ureteric tree, the number of branches and the nephron induced by each ureteric bud tip were obtained by counting. In addition, paraffin sections were stained with HE staining for morphological observation of nephrogenic zone and ureteric bud, while in order to reflect the density of the ureteric bud tips at nephrogenic zone, the distance between two neighboring ureteric bud tips was measured aided with the Claudin-7 immunohistochemical staining. Results The ureteric bud branching tree revealed that the initial bifid iterative branching formed the framework of renal medulla, the branching became complicated and dense in cortex and nephrogenic zone, while the distance between ureteric bud tips were also decreasing. The number of the nephron induced by each ureteric bud tip increased from one (E14. 5) to two (E17. 5), and occasionally to three. Conclusion Threedimeasional Visualization of ureteric bud branching tree reveals regional complication, suggesting molecules in different regions drive different branching patterns; While the density of the ureteric bud tips at nephrogenic zone increases corresponding to decreasing of thickness of the nephrogenic zone, and the disappearance of the ureteric bud tips after birth is also consistent with the gradual consumption of nephron progenitor cells.

In the research on cancer theranostics, most environment-sensitive drug delivery systems can only achieve unidirectional and irreversible responsive changes under pathological conditions, thereby improving the targeting effect and drug release performance of the delivery system. However, such irreversible changes pose potential safety hazards when the dynamically distributed delivery system returns to the blood circulation or transports to the normal physiological environment. Intelligent reversible drug delivery systems can respond to normal physiological and pathological microenvironments to achieve bidirectional and reversible structural changes. This feature will help to precisely control the drug release of the delivery system, prolong the blood circulation time, improve the targeting efficiency, and avoid the potential safety hazards of the irreversible drug delivery system. In this review, we describe the research progress of intelligent reversible drug delivery system from two main aspects: controlled drug release and prolonged blood circulation time/enhanced cellular internalization of drug.

OBJECTIVE: To investigate serum thyroid stimulating hormone (TSH) level and its changes with age in apparently healthy Chinese elderly population and analyze the differences between TSH levels detected using Roche and Snibe electrochemiluminescence immunoassay analyzers. METHODS: General clinical data and frozen fasting serum samples were collected from 5451 apparently healthy Chinese elderly individuals (> 60 years) from 10 centers in different geographic regions in China. Thyroid function indexes including TSH level were detected using Roche and Snibe electrochemiluminescence immunoassay analyzer, and the median (2.5% and 97.5% quantiles) TSH level was calculated. The variations of TSH level among the participants with geographic regions, gender, and age (with an interval of 5 years) were analyzed to determine the influence of these factors on TSH level. RESULTS: The reference ranges of serum TSH level established using Roche and Snibe electrochemiluminescence immunoassay analyzers were 0.42-9.47 mU/L and 0.36-7.98 mU/L, respectively, showing significant differences between the two methods (P < 0.001). The TSH levels measured at two centers in Western China were significantly higher than those at the other centers (P < 0.05). In elderly male population, serum TSH level tended to increase with age, which was not observed in elderly female population. At the age of 60-75 years, women generally had higher serum TSH level than men, but this difference was not observed in the population beyond 75 years. CONCLUSION In elderly population, serum TSH level can vary with geographic region, gender, and age, but there was no need for establishing specific reference ranges for these factors. The differences between different detection methods should be evaluated when interpreting the detection results of TSH level.
Aged ; Female ; Humans ; Male ; Middle Aged ; Asian People ; China ; Fasting ; Health Status ; Thyrotropin/blood*

PURPOSE: This study aims to elucidate the electrotaxis response of alveolar epithelial cells (AECs) in direct-current electric fields (EFs), explore the impact of EFs on the cell fate of AECs, and lay the foundation for future exploitation of EFs for the treatment of acute lung injury. METHODS: AECs were extracted from rat lung tissues using magnetic-activated cell sorting. To elucidate the electrotaxis responses of AECs, different voltages of EFs (0, 50, 100, and 200 mV/mm) were applied to two types of AECs, respectively. Cell migrations were recorded and trajectories were pooled to better demonstrate cellular activities through graphs. Cell directionality was calculated as the cosine value of the angle formed by the EF vector and cell migration. To further demonstrate the impact of EFs on the pulmonary tissue, the human bronchial epithelial cells transformed with Ad12-SV40 2B (BEAS-2B cells) were obtained and experimented under the same conditions as AECs. To determine the influence on cell fate, cells underwent electric stimulation were collected to perform Western blot analysis. RESULTS: The successful separation and culturing of AECs were confirmed through immunofluorescence staining. Compared with the control, AECs in EFs demonstrated a significant directionality in a voltage-dependent way. In general, type Ⅰ alveolar epithelial cells migrated faster than type Ⅱ alveolar epithelial cells, and under EFs, these two types of cells exhibited different response threshold. For type Ⅱ alveolar epithelial cells, only EFs at 200 mV/mm resulted a significant difference to the velocity, whereas for, EFs at both 100 mV/mm and 200 mV/mm gave rise to a significant difference. Western blotting suggested that EFs led to an increased expression of a AKT and myeloid leukemia 1 and a decreased expression of Bcl-2-associated X protein and Bcl-2-like protein 11. CONCLUSION EFs could guide and accelerate the directional migration of AECs and exert antiapoptotic effects, which indicated that EFs are important biophysical signals in the re-epithelialization of alveolar epithelium in lung injury.
Humans ; Rats ; Animals ; Alveolar Epithelial Cells ; Lung ; Lung Injury ; Cell Movement/physiology*

Objective: To observe the platinum drugs resistance effect of N-acetyltransferase 10 (NAT10) overexpression in breast cancer cell line and elucidate the underlining mechanisms. Methods: The experiment was divided into wild-type (MCF-7 wild-type cells without any treatment) group, NAT10 overexpression group (H-NAT10 plasmid transfected into MCF-7 cells) and NAT10 knockdown group (SH-NAT10 plasmid transfected into MCF-7 cells). The invasion was detected by Transwell array, the interaction between NAT10 and PARP1 was detected by co-immunoprecipitation. The impact of NAT10 overexpression or knockdown on the acetylation level of PARP1 and its half-life was also determined. Immunostaining and IP array were used to detect the recruitment of DNA damage repair protein by acetylated PARP1. Flow cytometry was used to detect the cell apoptosis. Results: Transwell invasion assay showed that the number of cell invasion was 483.00±46.90 in the NAT10 overexpression group, 469.00±40.50 in the NAT10 knockdown group, and 445.00±35.50 in the MCF-7 wild-type cells, and the differences were not statistically significant (P>0.05). In the presence of 10 μmol/L oxaliplatin, the number of cell invasion was 502.00±45.60 in the NAT10 overexpression group and 105.00±20.50 in the NAT10 knockdown group, both statistically significant (P<0.05) compared with 219.00±31.50 in wild-type cells. In the presence of 10 μmol/L oxaliplatin, NAT10 overexpression enhanced the binding of PARP1 to NAT10 compared with wild-type cells, whereas the use of the NAT10 inhibitor Remodelin inhibited the mutual binding of the two. Overexpression of NAT10 induced PARP1 acetylation followed by increased PARP1 binding to XRCC1, and knockdown of NAT10 expression reduced PARP1 binding to XRCC1. Overexpression of NAT10 enhanced PARP1 binding to LIG3, while knockdown of NAT10 expression decreased PARP1 binding to LIG3. In 10 μmol/L oxaliplatin-treated cells, the γH2AX expression level was 0.38±0.02 in NAT10 overexpressing cells and 1.36±0.15 in NAT10 knockdown cells, both statistically significant (P<0.05) compared with 1.00±0.00 in wild-type cells. In 10 μmol/L oxaliplatin treated cells, the apoptosis rate was (6.54±0.68)% in the NAT10 overexpression group and (12.98±2.54)% in the NAT10 knockdown group, both of which were statistically significant (P<0.05) compared with (9.67±0.37)% in wild-type cells. Conclusion: NAT10 overexpression enhances the binding of NAT10 to PARP1 and promotes the acetylation of PARP1, which in turn prolongs the half-life of PARP1, thus enhancing PARP1 recruitment of DNA damage repair related proteins to the damage sites, promoting DNA damage repair and ultimately the survival of breast cancer cells.
Breast Neoplasms/enzymology* ; Cell Line, Tumor ; Drug Resistance, Neoplasm ; Female ; Humans ; MCF-7 Cells ; N-Terminal Acetyltransferases/metabolism* ; Organoplatinum Compounds/pharmacology* ; Oxaliplatin/pharmacology* ; X-ray Repair Cross Complementing Protein 1

Objective Adult proximal tubule ( PT ) is not only the segment most frequently involved in acute renal tubule injury, but also the easiest to repair. It may be consistent with the rapid growth and differentiation mechanism of this segment during the development of the kidney, while the developing information is insufficient. Therefore, we three- dimensional visualized the developing PT to analysis its spatiotemporal morphogenesis. Methods The kidneys were obtained from mice at various developing time point, embryonic day ( E ), postnatal day ( P ). The volume density of Claudin-2 positive PT in the cortex was measured using a stereological method in paraffin sections. After image recording and alignment of the serial sections, the spatial courses of the developing PT were traced and visualized in three dimensions using computer-assisted program. The length of the developing PT was calculated at the same time. Results The volume density of PT in the cortex of PI mice was significantly higher than that in the embryonic stage. Then it experienced a decline ( P3, P5 ), an increase ( start at P7 ) to a stable adult level ( P28 ). The tubular tracing showed that the lengths of developing PT and the number of convolutions of their convoluted part increased with the maturation, but lower than that of adultin E14. 5, E17. 5 and P5 PT in E14. 5 and E17. 5 mice were similar to that of adult with respect to general spatial courses. They were, however, significantly different from adult in the initial direction of PT and the arrangement of the straight part of PT in the medullary rays. While, it was in P5 that the spatial pattern of some PT was gradually approaching to the adult model. Conclusion This study demonstrated that the development of PT was consistent with the kidney development in terms of its volume density in cortex, length and spatial course. It started at the S-shaped body, kept throughout the embryonic period and continued to postnatal, ended at kidney maturation ( P28 ).

OBJECTIVE: To investigate the correlation of platelet and coagulation function with blood stasis syndrome (BSS) in coronary heart disease (CHD). METHODS: The protocol for this meta-analysis was registered on PROSPERO (CRD42019129452). PubMed, Excerpta Medica Database (Embase), the Cochrane Library, and China National Knowledge Infrastructure (CNKI) were searched from inception to 1st June, 2020. Trials were considered eligible if they enrolled BSS and non-BSS (NBSS) patients with CHD and provided information on platelet and coagulation function. The platelet function, coagulation function, and fibrinolytic activity were compared between the BSS and NBSS groups. Forest plots were generated to show the SMDs or ESs with corresponding 95% CIs for each study. Subgroup analysis and sensitivity analysis were performed to explore potential sources of heterogeneity. RESULTS: The systematic search identified 1,583 articles. Thirty trials involving 10,323 patients were included in the meta-analysis. The results showed that mean platelet volume, platelet distribution width, platelet aggregation rate, platelet P selectin, fibrinogen, plasminogen activator inhibitor-1 (PAI-1), thromboxane B2 (TXB2), 6-keto-prostaglandin F1alpha (6-keto-PGF1 α), and TXB2/6-keto-PGF1 α were higher in the BSS group than in the NBSS group (P<0.05 or P<0.01). Activated partial thromboplastin time was lower in the BSS group than in the NBSS group in the acute phase of CHD (P<0.01). The R and K values in thromboelastography and tissue plasminogen activator (t-PA) and t-PA/PAI-1 were lower in the BSS group than in the NBSS group (all P<0.01). No difference was found in the results of platelet count, plateletcrit, maximum amplitude, von Willebrand factor, prothrombin time, thrombin time, international normalized ratio, etc. between groups. CONCLUSIONS Increased platelet function, hypercoagulability, and decreased fibrinolytic activity were found among CHD patients with BSS.
Blood Coagulation ; Blood Platelets ; Coronary Disease ; Humans ; Platelet Aggregation ; Tissue Plasminogen Activator

OBJECTIVETo investigate the pro-angiogenic effects of paeoniflorin (PF) in a vascular insufficiency model of zebrafish and in human umbilical vein endothelial cells (HUVECs).

METHODSIn vivo, the pro-angiogenic effects of PF were tested in a vascular insufficiency model in the Tg(fli-1:EGFP)y1 transgenic zebrafish. The 24 h post fertilization (hpf) embryos were pretreated with vascular endothelial growth factor (VEGF) receptor tyrosine kinase inhibitor II (VRI) for 3 h to establish the vascular insufficiency model and then post-treated with PF for 24 h. The formation of intersegmental vessels (ISVs) was observed with a fluorescence microscope. The mRNA expression of fms-like tyrosine kinase-1 (flt-1), kinase insert domain receptor (kdr), kinase insert domain receptor like (kdrl) and von Willebrand factor (vWF) were analyzed by real-time polymerase chain reaction (PCR). In vitro, the pro-angiogenic effects of PF were observed in HUVECs in which cell proliferation, migration and tube formation were assessed.

RESULTSPF (6.25-100 μmol/L) could rescue VRI-induced blood vessel loss in zebrafish and PF (25-100 μmol/L), thereby restoring the mRNA expressions of flt-1, kdr, kdrl and vWF, which were down-regulated by VRI treatment. In addition, PF (0.001-0.03 μmol/L) could promote the proliferation of HUVECs while PF stimulated HUVECs migration at 1.0-10 μmol/L and tube formation at 0.3 μmol/L.

CONCLUSIONPF could promote angiogenesis in a vascular insufficiency model of zebrafish in vivo and in HUVECs in vitro.

Angiogenesis Inducing Agents ; pharmacology ; therapeutic use ; Animals ; Animals, Genetically Modified ; Cells, Cultured ; Disease Models, Animal ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Embryo, Nonmammalian ; Glucosides ; pharmacology ; therapeutic use ; Human Umbilical Vein Endothelial Cells ; drug effects ; physiology ; Humans ; Monoterpenes ; pharmacology ; therapeutic use ; Neovascularization, Physiologic ; drug effects ; Phytotherapy ; Vascular Diseases ; drug therapy ; pathology ; Zebrafish

·AIM: To study the mechanism and effect of Qingxuan decoction on evaporative dry eye in rabbit model. ·METHODS: Totally 25 healthy male Japanese white rabbits were randomly divided into 5 groups: control group, model group, western medicine group, high dose of Qingxuan decoction group, low dose of Qingxuan decoction group. The blank control group did not do any treatment. The improved dry eye model of rabbit was prepared by the improved method of glandular burning of the eyelid plate. The high and low dose group were given daily 27. 2mg/kg, 6. 8mg/kg Qingxuan decoction by gavage. The model group was intragastric with the same amount of normal saline every day. The western medicine group with tobramycin and dexamethasone ophthalmic ointment 1 drops, once a day. The treatment were administered continuously for 28d. At 14d before the experiment, 7d before the experiment, 7d after the model, and 14d after the model, all the rabbits were tested by Schirmer Ⅰ test ( SⅠt) and break-up time (BUT). On the 15d after modeling, the animals were sacrificed by excessive anaesthesia. Rabbit ocular surface tissue sections were prepared. Hematoxylin - eosin staining method was used to observe the corneal morphological changes in each group. The concentrations of TNF-α, IL-1 and IL-6 in the ocular surface of rabbits were detected by ELISA. ·RESULTS: (1) BUT, SⅠt: 7d after the model had been prepared, BUT and SⅠt of the model group and the western medicine group, high dose and low dose of Qingxuan decoction group was improved ( P< 0. 05 );Those of western medicine group, high dose and low dose of Qingxuan Decoction group compared with the model group, were significantly different (P<0. 05). (2) TNF-α, IL-1, IL-6: The ELISA assay showed that TNF-α and IL-1, IL-6 concentration in the model group rabbits was significantly higher than those of the control group, TNF-α and IL-1, IL-6 concentration in western medicine group and high dose group of rabbits was significantly lower than those in the model group, the differences were statistically significant (P<0. 05), and in high dose group the effect was better than that of Western medicine group. ( 3 ) Histopathological examination: on the 14d after the model, the corneal epithelium in the blank control group was stratified well. The cells in the base were columnar, near the surface, the cornea epithelium showed a squamous change. Conjunctiva showed complete epithelial layer and subconjunctival tissue layer, and goblet cells arranged closely. The number of corneal epithelial cells in model group was reduced or even stripped, and the matrix layer was disorder; Irregular loss of conjunctival epithelial cell layer and a large decrease in goblet cells. The corneal morphology of the rabbits in the western medicine group and the high dose group was close to the normal group, and the number of conjunctival goblet cells was not significantly different from that in the blank control group. ·CONCLUSION: The expression of Qingxuan decoction can inhibit the inflammatory reaction through down -regulation of TNF-α and IL-1, IL-6 and in evaporative dry eye rabbit cornea and conjunctiva, so as to improve the ocular symptoms, increase tear secretion, prolong the time of BUT.