Gastric cancer-derived mesenchymal stem cells regulate the M2 polarization of macrophages within gastric cancer microenvironment via JAK2/STAT3 signaling pathway / 中华肿瘤杂志

Objective: To investigate the role and mechanism of tumor-derived mesenchymal stem cells in regulating the M2 polarization of macrophages within gastric cancer microenvironment. Methods: Gastric cancer tissues and the adjacent non-cancerous tissues were collected from patients underwent gastric cancer resection in the First People's Hospital of Lianyungang during 2018. In our study, THP-1-differentiated macrophages were co-cultured with gastric cancer-derived mesenchymal stem cells (GC-MSCs). Then, the M2 subtype-related gene, the markers expressed on cell surface and the cytokine profile were analyzed by real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR), flow cytometry and Luminex liquid chip, respectively. The key cytokines mediating the inducing effect of GC-MSCs on macrophage polarization into the M2 subtype were detected and screened by Luminex liquid chip, which were further confirmed by the neutralizing antibody test. The expressions of macrophage proteins involved in M2 polarization-related signaling pathways under the different co-culture conditions of GC-MSCs were detected by western blot. Results: In Mac+ GC-MSC-culture medium (CM) group, the expression levels of Ym-1 and Fizz-1 (1.53±0.32 and 13.22±1.05, respectively), which are markers for M2 subtype, were both significantly higher than those of Mac group (1.00±0.05 and 1.21±0.38, respectively, P<0.05). The level of iNOS in Mac+ GC-MSC-CM group (0.60±0.41) was significantly lower than that of Mac group (1.06±0.38, P=0.023). In Mac+ GC-MSC-Transwell (TW) group, the expression levels of Ym-1 and Fizz-1 (1.47±0.09 and 13.16±2.77, respectively) were both significantly higher than those of Mac group (1.00±0.05 and 1.21±0.38, respectively, P<0.05). The level of iNOS in Mac+ GC-MSC-CM group (0.56±0.03) was significantly lower than that of Mac group (1.06±0.38, P=0.026). The ratios of CD163(+) /CD204(+) cells in Mac+ GC-MSC-CM and Mac+ GC-MSC-TW groups (3.80% and 4.40%, respectively) were both remarkably higher than that of Mac group (0.60%, P<0.05). The expression levels of IL-10, IL-6, MCP-1 and VEGF in Mac+ GC-MSC-CM group were (592.60±87.52), (1 346.80±64.70), (11 256.00±29.03) and (1 463.90±66.67) pg/ml, respectively, which were significantly higher than those of Mac group [(41.03±2.59), (17.35±1.79), (5 213.30±523.71) and (267.12±12.06) pg/ml, respectively, P<0.05]. The levels of TNF-α, IP-10, RANTES and MIP-1α were (95.57±9.34), (410.48±40.68), (6 967.30±1.29) and (1 538.70±283.04) pg/ml, which were significantly lower than those of Mac group [(138.01±24.31, (1 298.60±310.50), (14 631.00±4.21) and (6 633.20±1.47) pg/ml, respectively, P<0.05]. The levels of IL-6 and IL-8 in GC-MSCs [(11 185.02±2.82) and (12 718.03±370.17) pg/ml, respectively] were both strikingly higher than those of MSCs from adjacent non-cancerous gastric cancer tissues [(270.71±59.38) and (106.04±32.84) pg/ml, repectively, P<0.05]. The ratios of CD86(+) cells in Mac+ IL-6-blocked-GC-MSC-CM and Mac+ IL-8-blocked-GC-MSC-CM groups (28.80% and 31.40%, respectively) were both higher than that of Mac+ GC-MSC-CM group (24.70%). Compared to Mac+ GC-MSC-CM group (13.70%), the ratios of CD204(+) cells in Mac+ IL-6-blocked-GC-MSC-CM and Mac+ IL-8-blocked-GC-MSC-CM groups (9.90% and 8.70%, separately) were reduced. The expression levels of p-JAK2 and p-STAT3, which are proteins of macrophage M2 polarization-related signaling pathway, in Mac+ GC-MSC-CM group (0.86±0.01 and 1.08±0.01, respectively) were significantly higher than those of Mac group (0.50±0.01 and 0.82±0.01, respectively, P<0.05). The expression levels of p-JAK2 in Mac+ IL-6-blocked-GC-MSC-CM group (0.47±0.02) were significantly lower those that of Mac+ GC-MSC-CM group (0.86±0.01, P<0.05). The expression levels of p-JAK2 and p-STAT3 in Mac+ IL-8-blocked-GC-MSC-CM group (0.50±0.01 and 0.85±0.01, respectively) were both significantly lower than those of Mac+ GC-MSC-CM group (0.86±0.01 and 1.08±0.01, P<0.05). The expression levels of p-JAK2 and p-STAT3 in Mac+ IL-6/IL-8-blocked-GC-MSC-CM group (0.37±0.01 and 0.65±0.01, respectively) were both significantly lower than those of Mac+ GC-MSC-CM group (0.86±0.01 and 1.08±0.01, P<0.05). Conclusion: GC-MSCs promote the activation of JAK2/STAT3 signaling pathway in macrophages via high secretions of IL-6 and IL-8, which subsequently induce the macrophage polarization into a pro-tumor M2 subtype within gastric cancer microenvironment.

The Liver Injury and Coagulation Dysfunction in the Patients with Severe/Critical COVID-19 / 中国实验血液学杂志

METHODS@#The clinical data of 53 COVID-19 patients were collected from a single center in Wuhan from February 8, 2020 to March 25, 2020. The patients were divided into severe type group (38 patients) and critical type group (15 patients). The clinical characteristics, indexes of liver function, coagulation function and inflammatory markers were analyzed retrospectively. According to the degree of abnormal liver function in the process of diagnosis and treatment, the patients were divided into three groups: combined liver injury, mild abnormal liver function and normal liver function group. Statistical analysis was performed by using Student t test, Mann-Whitney U test, Kruskal-Wallis test and Chi-square test.@*RESULTS@#Among the 53 patients, 29 were male (54.7%) and 24 were female (45.3%), the median age was 57(27－80) years old. The time from onset to admission was (11.5±7.7) days. The levels of AST, TBIL, DBIL, ALP, GGT, LDH, D-dimer, PCT and hsCRP in critical patients were higher than those in severe patients (P<0.05). The levels of Alb in critical patients was lower than those in severe patients (P<0.05). Among the 53 patients, 34 (64%) patients showed abnormal elevation of ALT, AST or TBIL, while 4 (7.5%) patients showed the criteria of COVID-19 with liver injury. After the patients were grouping according to the degree of liver dysfunction, the levels of ALP, GGT and D-dimer of the patients in the liver injury group were significantly higher than those in the normal liver function group, D-dimer levels of the patients in the liver injury group was significantly higher than those in the mild abnormal liver function group, while the levels of ALP and GGT in the mild abnormal liver function group were significantly higher than those in the normal liver function group, and the differences were statistically significant(P<0.05).@*CONCLUSION@#In this group, the patients with COVID-19 severe/critical type have a certain proportion of liver injury accompanied by significantly increased D-dimer levels, critical type patients have more severe liver function and coagulation dysfunction, which may promote the progression of COVID-19.

The influence of M2 macrophages on tumor-promoting effect of gastric cancer-derived mesenchymal stem cells in gastric cancer microenvironment / 重庆医学

Objective To investigate the effect of M2 macrophages resident in gastric cancer microenvironment on the tumor-promoting effect of human gastric cancer-derived mesenchymal stem cells (GC-MSCs).Methods Macrophages in BALB/c mice were depleted by using clodronate liposomes.The tumor volumes and weights in nude mice co-injected with GC-MSCs and BGC-823 with and without macrophage depletion were recorded.Tumor tissues of nude mice and gastric cancer patients were collected,and M2 macrophage-associated genes and proteins were detected by RT-PCR and western blot.Furthermore,the regulating effect of GC-MSCs on macrophage polarization to M2-subtype was validated in the co-culture experiment in vitro.Results Tumor growth in GC-MSCs co-injected mice was significantly inhibited by macrophage depletion (P=0.009).Results of RT-PCR and western blot showed that the transcription and expression of M2 macrophage-associated proteins were significantly higher in tumor tissues from GC-MSCs co-injected mice than those in the control group.Moreover,the transcription and expression levels of M2 macrophage-associated proteins were also high-er in gastric cancer tissues than those in the corresponding adjacent normal tissues.After co-culture with GC-MSCs directly,the expressions of M2 macrophage-associated proteins were significantly up-regulated in THP-1-derived macrophages.Conclusion M2 macrophages in gastric cancer microenvironment might play a critical role in the tumor-promoting effect of GC-MSCs.

Study progress in capsid protein VP1 of enterovirus 71 / 中华实用儿科临床杂志

Human enterovirus 71 (EV71) is one of the major causative agent of hand,foot and mouth disease (HFMD) in infant.Clinical studies find that EV71 infection can cause a variety of clinical manifestations,from mild HFMD to fatal neurogenic pulmonary edema and even death,but the reasons remain unclear.The capsid protein VP1 of EV71 plays a key role in the processes of viral recognizing,binding and entering into the targeted cells and viral particles assembling.VP1 variation is a major determinant to EV71 fitness and immunogenicity.This study reviews the research progress of the structure,functions and associated antiviral vaccines and drugs of VP1,which further provides a theoretical basis for developing new and more effective antiviral vaccine and drugs.

Curcumin induces apoptosis and protective autophagy in human gastric cancer cells with different degree of differentiation / 中华肿瘤杂志

Objective@#To investigate the effect of curcumin on the apoptosis and autophagy of human gastric cancer cells with different degree of differentiation.@*Methods@#Gastric cancer cell lines BGC-823 and MKN-28 were treated with curcumin at different concentrations. The effect of curcumin on cell proliferation was measured by MTT assay. Apoptosis was assessed by flow cytometry. Autophagy status was analyzed by acridine orange staining. The expression levels of apoptotic and autophagy-related proteins were detected by Western blot.@*Results@#The cell viability of BGC-823 and MKN-28 was inhibited by curcumin in a time- and dose-dependent manner. At 48 h after treatment, the IC50 value of BGC-823 (15.18 μmol/L) was close to that of MKN-28 (15.84 μmol/L), and the difference was not statistically significant (P=0.513). Meanwhile, flow cytometry showed that curcumin induced the apoptosis of gastric cancer cells in a dose-dependent manner. Western blot results showed that the expression of pro-apoptotic proteins bax, active-caspase-3 and active-caspase-9 was significantly increased in BGC-823 and MKN-28 cells, whereas that of the anti-apoptotic protein bcl-2 was strikingly reduced. In addition, the formation of acidic vesicular organelles in cytoplasm, conversion of LC3-Ⅰ to LC3-Ⅱ and increased levels of autophagy-related proteins Beclin1, Atg7 and Atg5-Atg12 were observed in curcumin-treated cells. Moreover, activation of PI3K/Akt/mTOR signaling pathway was also significantly suppressed after curcumin treatment. Blocking autophagy by adding the autophagy inhibitor 3-methyladenine (3-MA) significantly promoted the apoptotic cell death induced by curcumin.@*Conclusions@#Curcumin induces apoptosis and protective autophagy in human gastric cancer cells in vitro. Curcumin combined with autophagy inhibitor may provide a more effective strategy for its clinical application.

Detection of membrane neutrophilic alkaline phosphatase by flow cytometry in diagnosis of ;bloodstream infection / 中华临床感染病杂志

Objective To evaluate the detection of membrane neutrophilic alkaline phosphatase ( mNAP) by flow cytometry in diagnosis of bloodstream infection .Methods A total of 298 patients with suspected bloodstream infections admitted in the First People ’ s Hospital of Lianyungang during June 2013 and October 2014 were enrolled;80 healthy subjects in physical examination center were also enrolled as the control group.Bloodstream infection was diagnosed by blood culture and mNAP was detected by flow cytometry.Serum levels of procalcitonin (PCT) and C-reactive protein (CRP) were detected by electro-chemiluminescence (ECL) and immune scatter turbidimetry , respectively.The value of mNAP, PCT and CRP in diagnosing bloodstream infection was determined by receiver operating characteristic ( ROC) curve. Results Among 298 patients, 109 were confirmed with bloodstream infections , including 43 patients with Gram-positive bacterial infections and 66 with Gram-negative bacterial infections .The median levels of CRP , PCT and mNAP in bloodstream infection group were 138.71 mg/L, 7.04 ng/mL and 13 929 AB/c, which were significantly higher than those in healthy control group (1.50 mg/L, 0.12 ng/mL, 1 831 AB/c;U=5.00, 48.50 and 65.01, P<0.01).The expression of mNAP in Gram-positive bacterial infection group was 9 598 ( 6 064-11 643 ) AB/c, which was significantly lower than that in Gram-negative bacterial infection group [16 512 (11 654-22 001) AB/c] (U=250.00, P<0.01).ROC curve analysis showed that, the areas under the curve (AUCs) of mNAP, PCT and CRP in diagnosing bloodstream infection were 0.987, 0.962 and 0.901.When 4 578AB/c, 0.90 ng/mL and 13.50mg/L were taken as optimal cut-off values, the sensitivities of mNAP, PCT and CRP in diagnosis of bloodstream infection were 95.8%, 93.0%and 90.3%; the specificities were 97.8%, 95.6% and 85.5%, respectively.Conclusion Among mNAP, PCT and CRP, mNAP is of the highest value in diagnosing bloodstream infection , and may be used as a biomarker for clinical diagnosis of bloodstream infection .

Differential expression in ACE2, Ang(1-7) and Mas receptor during progression of liver fibrosis in a rat model / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To investigate the changes in expression of the ACE2/Ang(1-7)/Mas receptor axis' components that occur during progression of liver fibrosis using a rat model system.</p><p><b>METHODS</b>Thirty-six adult male Wistar rats, were randomly assigned to groups of normal control (n = 6; no manipulation) and liver fibrosis (n = 30; given a subcutaneous injection of 40% chronic carbon tetrachloride (CCl4)). At post-injection days 15, 30, 45, 60 and 75, 1 control rat and 6 modeled rats were sacrificed for analysis. Histopathological analysis of liver tissue was performed with hematoxylin-eosin and rapid Masson staining. Protein expression level of Ang(1-7) was determined by enzyme-linked immunosorbent assay, and of ACE2 and Mas receptor was evaluated by immunohistochemistry. Real-time PCR was used to measure the mRNA expression levels of ACE2 and Mas receptor.</p><p><b>RESULTS</b>The expression levels of ACE2, Ang(1-7) and Mas receptor showed a statistically significant upward trend that followed the progression of fibrosis up to post-injection day 60 (P less than 0.01), but the significant increase was not seen from day 60 to day 75.</p><p><b>CONCLUSION</b>Each component of the ACE2/Ang(1-7)/Mas receptor axis shows differential expression during the development of liver fibrosis and may contribute to disease progression.</p>

Study on bioactive constituents of the South China Sea soft coral Scleronephthya sp / 第二军医大学学报

Objective: To investigate the bioactive constituents of coral Scleronephthya sp. collected from the South China Sea. Methods: The compounds were isolated and purified using repeated column chromatographies on Sephadex LH-20, normal- and reversed-phase silica gels, and RP-HPLC. The structures of the compounds were elucidated based on the detailed spectroscopic analysis in combination with reported data. The in vitro antimicrobial activity of these compounds were assessed by an agar diffusion test. Results and conclusion: Three 5α, 8α-epidioxysterols, two alkyl glycerol ethers and 1, 2-O-alkyl glycerol were isolated from Scleronephthya sp. ; these compounds displayed different levels of antifungal and antibacterial activities in bioassay in vitro. Compound 5 inhibited the growth of Gram-negative bacterium Escherichia coli and the Gram-positive bacterium Bacillus megaterium. Compound 6 showed marked antifungal activity against Microbotryum violaceum. Compounds 1-4 demonstrated a weak antimicrobial activity.