AG490 regulates JAK2/STAT3 pathway in M2-like macrophages via promoting the proliferation of gastric cancer cells and inhibiting apoptosis / 西安交通大学学报(医学版)

【Objective】 To investigate the regulatory effect of JAK2/STAT3 signaling pathway inhibitor AG490 on the functional phenotype of M2-like macrophages and its effects on proliferation, apoptosis, migration, and invasion of gastric cancer cells. 【Methods】 Human mononuclear cell line THP-1 was induced to differentiate into M0 macrophages by PMA in vitro. The M1-like phenotype was induced by LPS and IFN-γ, and M2-like phenotype was induced by IL-4 and IL-13, respectively, and identified by immunofluorescence labeling CD68, CD86 and CD206. The mRNA expressions of CD163, Arg1, CCL22, PPARγ, IL-10, IL-20 and TNF-α were determined by RT-qPCR. The expressions of key proteins in JAK2/STAT3 signaling pathway were detected by Western blotting. M2-like macrophages were treated with JAK2/STAT3 inhibitor (AG490) to observe the expression level of marker genes for M2 like phenotype. Macrophages were co-cultured with gastric cancer cells, and the effects of the macrophages on proliferation, migration, invasion, and apoptosis of gastric cancer cells were detected by CCK-8 method, healing assay, transwell intracellular Matrigel invasion assay, and flow cytometry. The xenograft tumor model of MKN45 gastric cancer in nude mice was prepared, and the tumor size and quality were observed for 20 days after the model was established. 【Results】 THP-1 cells were induced into M1-like macrophages and M2-like macrophages. M1-like marker (CD86) and M2-like marker (CD206) were identified by flow cytometry. The P38MAPK, JAK2, p-STAT3/STAT3 protein levels of M2-like macrophages treated with AG490 were significantly reduced. The mRNA expression levels of Arg1, CCL22, PPARγ and IL-10 were significantly reduced in the group of M2-like macrophages treated with AG490. Co-culture of M2-like macrophages with gastric cancer cells could promote gastric cancer cell viability, increase migration and invasion ability, and inhibit apoptosis. When the group of M2-like macrophages treated with AG490 was co-cultured with gastric cancer cells, the proliferation activity of MKN-45 cells and MGC823 was significantly lower than that in M2 group (1.047±0.062 vs. 1.426±0.076, 1.149±0.006 vs. 1.301±0.015). Compared to M2 group, the migration (100.0%±5.73% vs. 72%±3.85%) and invasion ability (100.0%±7.40% vs. 60%±6.54%) of MGC823 gastric cancer cells in AG490 treatment group were significantly reduced. The apoptosis rate of MGC823 cells in the AG490 treated group was significantly higher than that in M2 group (27.51%±0.70% vs. 20.82%±0.92%). In the nude mouse xenograft tumor model, the volume and weight of the transplanted tumor collected at day 20 were significantly lower in AG490 treated group than in M2 group (736.04±182.34 vs. 1 080.5±250.57)mm3, (0.64±0.11 vs. 0.87±0.17)g. 【Conclusion】 AG490 downregulates the activation level of JAK2/STAT3 signaling pathway in M2-like macrophages, inhibits M2-type polarization, partially reverses the cancer-promoting function of M2-like macrophages, inhibits proliferation, migration and invasion of gastric cancer cells, and induces apoptosis. The JAK2/STAT3 signaling pathway can be further studied as a potential therapeutic target for gastric cancer.

Phenotypic changes of spleen macrophages in advanced gastric cancer and their effects on proliferation, invasion and migration of gastric cancer cells / 西安交通大学学报(医学版)

Objective: To investigate the phenotypic changes of splenic macrophages in advanced gastric cancer and the effects of co-culture with M1 and M2 macrophages on gastric cancer cells in order to explore the role of the spleen in the development of gastric cancer. Methods: We collected the fresh surgically removed spleens from 15 patients who had undergone advanced proximal gastrectomy combined with splenectomy in our hospital from March 2015 to May 2017. Spleen macrophages were isolated from these spleen tissues and the macrophage phenotypes were detected by flow cytometry. Monocytes were isolated from fresh blood of healthy volunteers using a fully automated bead extraction system; the identified monocytes were given 10 ng/mL of GM-CSF and M-CSF to be induced into M1-type macrophages and M2-type macrophages, respectively. The phenotypes of macrophages were identified by flow cytometry using CD16 and CD163; the gastric cancer cells SGC-7901 and AGS were co-cultured with M1, M2 macrophages and monocytes using Transwell co-culture method; trypan blue staining was used to detect the proliferative, invasive and migratory abilities of the cells. Results: The proportion of type M2 macrophages in the spleen of patients with advanced gastric cancer was increased while the proportion of type M1 macrophages decreased. The mononuclear cells were induced into M1 and M2 macrophages by culture with GM-CSFand M-CSF, and the positive rate of monocytes was (95.46±4.21)% and (94.67±4.97)%, respectively; M2 macrophages and monocytes could promote the invasion and migration of gastric cancer cells, while M1 macrophages inhibited the invasion and migration of gastric cancer cells. Conclusion: This study identified the proportions of different macrophages in spleen tissues of patients with advanced gastric cancer, and confirmed that the different types of macrophages on gastric cancer cells have different effects. M2 macrophages promoted gastric cancer cells, while M1 macrophages showed the effect of inhibiting gastric cancer cells. Our research further revealed the role of the spleen in the progression of gastric cancer and provided experimental research basis for formulating a treatment plan for advanced gastric cancer.

Effect of Compound Duzhenggan Jiegu Plaster for External Use on the Healing of Closed Distal Radius Frac-ture / 中国药房

OBJECTIVE:To investigate the effect of Compound duzhenggan jiegu plaster for external use on the healing of closed distal radius fracture. METHODS:68 cases of closed distal radial fracture were randomly assigned into treatment group(34 cases) and control group (34 cases). Treatment group was treated with Compound duzhenggan jiegu plaster for external use,and control group was treated with Dieda pills for external use. The changes of X-ray score,BMD and serum ALP content were com-pared between 2 groups after 30 d and 60 d treatment. RESULTS:In 2 groups,there were statistically significant difference in X-ray score,BMD and serum ALP content between 30 d and 60 d treatment(P<0.01). After 60 d treatment,there were statistical-ly significant difference in X-ray score,BMD and serum ALP content between 2 groups(P<0.01 or P<0.05). CONCLUSIONS:Compound duzhenggan jiegu plaster for external use in the treatment of closed radius distal fracture can promote the increase of BMD and serum ALP content,improve fracture repair and healing,shorten the healing time. The mechanism should be further re-searched.

Application of a lipid emulsion for parenteral nutrition support in intensive care patients following gastrointestinal surgeries / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the effect of parenteral nutrition support with a lipid emulsion formulation (containing soybean oil, medium chain triglycerides, olive oil, and fish oil [SMOF]) in intensive care patients following major gastrointestinal surgeries.</p><p><b>METHODS</b>According to a randomized, prospective and case-controlled design, 72 intensive care patients following major gastrointestinal surgeries between January and December, 2014 were randomized equally into SMOF group and control group to receive parenteral nutrition support with SMOF and medium or long chain lipid emulsion, respectively. Before and at 4 and 9 days after commencement of parenteral nutrition support, the patients were examined for alanine aminotransferase (ALT), total bilirubin (TBIL), albumin (propagated), C-reactive protein (CRP), interleukin 6 (IL-6), and endotoxin levels. The patients' average length of stay in intensive care unit (ICU), the days of using antibiotics, and the incidence rate of postoperative complication were recorded.</p><p><b>RESULTS</b>On day 4 postoperatively, the levels of CRP and IL-6 were significantly lower in SMOF group than in the control group (t=2.669 and 2.676, respectively; P<0.05), and on day 9, the patients in SMOF group showed significantly lower levels of ALT, TBIL, CRP and IL-6 (t=2.487, 3.497, 3.762, 2.180, respectively; P<0.05) than the control group, but ALB and endotoxin levels remained comparable between the two groups. The average length of stay in ICU and the days of using antibiotics were significantly shorter in SMOF group than in the control group (t=2.94 and 2.17, respectively; P<0.05); SMOF group showed a lower incidence of postoperative infections than the control group, but the difference was not statistically significant (χ² =1.047, P>0.05).</p><p><b>CONCLUSION</b>For intensive care patients following major gastrointestinal surgeries, postoperative parenteral nutrition support with SMOF can effectively reduce the release of inflammatory mediators, protect important visceral functions, reduce postoperative complications, shorten the length of ICU stay, and improve the prognosis of the patients.</p>

Effect of bafilomycin A1 on proliferation and oxaliplatin sensitivity in gastric cancer MGC-803 cells / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the effect of bafilomycin A1 (BAF) on the cell proliferation, invasiveness, apoptosis, and oxaliplatin sensitivity in gastric cancer MGC-803 cells.</p><p><b>METHODS</b>MGC-803 cells were divided into control group, BAF group, oxaliplatin group, and BAFµ oxaliplatin group. MTT assay and plate clone formation assay were used to assess the viability and colony forming ability of the cells after the treatments. The expression of nucleosomes in the cells was examined with ELISA. The cell migration and invasion after the treatments were evaluated. Western blotting was performed to detect the expression of Bcl-2 and Bax in the treated cells, and scanning electron microscopy, immunohistochemistry and Western blotting were employed to to observe the cell autophagy.</p><p><b>RESULTS</b>Compared with the control cells, the cells treated with BAF showed a substantial decrease in autophagosome accumulation with attenuated cell proliferation, migration and invasion. Compared with cells treated with oxaliplatin alone, the cells treated with both BAF and oxaliplatin showed significantly lowered autophagosome accumulation, suppressed cell proliferation, migration and invasion, increased cell apoptosis, increased Bax expression and lowered Bcl-2 expression.</p><p><b>CONCLUSION</b>BAF can inhibit the proliferation and invasiveness of MGC-803 cells, promote cell apoptosis by inhibiting autophagy, and enhances the sensitivity of the cells to oxaliplatin.</p>

Clinical significance of UbcH10 expression in gastric cancer / 中华普通外科杂志

Objective To investigate the expression and significance of UbcH10 in gastric cancer.Methods The expression of UbcH10 in 117 cases of gastric carcinoma tissues,who were followed up postoperatively,were detected by using immunohistochemistry; The expression among different histological grades and clinical stages are analyzed by statistics; Real-time polymerase chain reaction (Rt PCR) and Western blot method were used to detect the expressions of UbcH10 mRNA and protein in 43 cases of gastric carcinoma tissues and the cancer adjacent normal tissues.Results In paraffin fixed blocks of gastric cancer,positive rate of UbcH10 protein expression was higher than normal gastric tissues (83％ vs 55％,Rs =-0.397,P＜0.01).The difference of UbcH10 expression was not significant in gender,age,tumor site and size of the tumor(Rs =0.369,-0.378,-0.276,0.421,P ＜ 0.05),while it was significantly correlated to tumor differentiation,invasive depth,lymphatic metastasis and clinical stage (Rs =0.369,-0.378,-0.276,0.421,P ＜0.05).UbcH10 and Ki-67 protein and HER-2 expression were in significantly positive correlation (Rs =0.834,Rs =0.281,P ＜ 0.01).UbcH10 mRNA and protein expression in fresh gastric carcinoma tissues was higher than that in their corresponding adjacent normal tissues(ACt =32.2±5.19,4.69 ±1.26,t =26.8,P＜0.01),(0.74±0.45,0.11 ±0.04,t =8.09,P＜0.01).The -5 year patient's survival rate in UbcH10 highly-expressed gastric cancer was higher compared to those with UbcH10 low-expression(x2 =13.116,P ＜0.01).Conclusions UbcH10 is upregulated in gastric cancer,and expression of UbcH10 is closely related to the prognosis of patients with gastric cancer.

Application of nickel titanium compression anastomosis clip in gastrointestinal anastomosis / 中华消化外科杂志

Objective To investigate the efficacy and safety of nickel titanium compression anastomosis clip (CAC) in gastrointestinal anastomosis.Methods The clinical data of 413 patients who received gastrointestinal anastomosis with nickel titanium CAC or curved stapler at the First Affiliated Hospital of Fujian Medical University from January 2007 to December 2011 were retrospectively analyzed.All the patients were divided into the CAC group (227 patients) and the curved stapler group (186 patients).The time for gastrointestinal anastomosis,anastomotic bleeding,complications,condition of the incision sites,the first postoperative flatus and time for the excretion of the CAC between the 2 groups were compared.All data were analyzed using the t test or chisquare test.Results The time for gastrointestinal anastomosis of the CAC group was (8.3 ± 2.5) minutes,which was significantly shorter than (18.7 ± 3.6) minutes of the curved stapler group (t =5.74,P ＜ 0.05).There were no significant difference in the time of postoperative flatus,duration of hospital stay and costs between the 2 groups (t =0.27,1.32,3.11,P ＞ 0.05).No patients in the CAC group had anastomotic bleeding,while the incidence of anastomotic bleeding of the curved stapler group was 3.23 ％ (6/186),with a significant difference between the 2 groups (x2=7.43,P ＜ 0.05).The incidences of gastric emptying dysfunction of the CAC group and the curved stapler group were 0.88％ (2/227) and 1.08％ (2/186),with no significant difference between the 2 groups (x2=0.04,P ＞ 0.05).The incidences of postoperative anastomotic stricture and anastomositis were 0 (0/227) and 11.01％ (25/227) in the CAC group,and 2.15％ (4/186) and 21.50％ (40/186) in the curved stapler group,with significant differences between the 2 groups (x2 =4.93,8.49,P ＜ 0.05).No anastomotic fistula was detected in the 2 groups.The CAC was excreted at postoperative day 12-26.Conclusion CAC is safe and effective for gastrointestinal anastomosis.

Impact of splenectomy on the prognosis and immune function of patients with end stage carcinoma of the esophagogastric junction / 中华消化外科杂志

Objective To investigate the impact of splenectomy on the prognosis and immune function of patients with end stage carcinoma of the esophagogastric.Methods The clinical data of 61 patients with end stage carcinoma of the esophagogastric junction who were admitted to the First Affiliated Hospital of Fujian Medical University from January 2007 to December 2010 were retrospectively analyzed.All the patients were divided into splenectomy group (22 patients) and spleen preservation group (39 patients).The immune function before operation,on the 10th day and the 6th month after operation was detected.The intra-and postoperative conditions of the patients of the 2 groups were compared.All data were analyzed using the independent sample t test or chisquare test.Results The levels of IgA,IgG,IgM,CD3 and CD4 in the splenectomy group at the 10th day after operation were significantly higher than those before operation (t =2.55,3.33,3.40,2.92,2.10,P ＜ 0.05).The levels of IgA,IgG,IgM,CD3,CD4,the ratio of CD4/CD8 of the spleen preservation group at the 10th day after operation were significantly higher than those before operation (t =3.35,5.29,3.33,2.60,3.53,3.12,P ＜0.05).The levels of IgA,IgG,IgM,CD3,CD4 and the ratio of CD4/CD8 in the splenectomy group at postoperative month 6 were significantly lower than those at postoperative day 10 (t ＝2.75,4.40,3.06,2.51,2.24,2.29,P ＜ 0.05).The levels of IgA,IgG,IgM,CD4,CD8 and the ratio of CD4/CD8 of the spleen preservation group were significantly higher than those of the splenectomy group,while the level of CD8 in the spleen preservation group was significantly lower than that in the splenectomy group (t =1.70,2.10,2.70,2.16,2.13,2.83,P ＜ 0.05).The operation time of the splenectomy group was (152 ± 26) minutes,which was significantly longer than (130 ± 24) minutes of the spleen preservation group (t ＝ 3.42,P ＜ 0.05).There were no significant differences in the operative blood loss,incidence of postoperative infection,median survival time,1-year survival rate between the 2 groups (t =1.38,x2 =0.78,1.22,2.51,P ＞ 0.05).Conclusion Palliative gastrectomy could reverse the immune function of spleen by decreasing tumor burden for patients with end stage carcinoma of the esophagogastric junction who can not be treated by radical resection.Spleen preservation may have positive significance for the immune function and prognosis of patients with end stage carcinoma of the esophagogastric junction.

Clinical significance of Sipa1 expression in gastric cancer / 中国肿瘤临床

Objective:To measure the expression of signal-induced proliferation-associated gene 1 (Sipa1) in gastric cancer and to determine its association with the clinicopathological characteristics and prognosis of patients with gastric carcinoma. Methods:The Sipa1 mRNA and Sipa1 protein expression levels in 43 fresh gastric carcinoma tissues and adjacent normal tissues were measured by real-time PCR and Western blot analysis. The Sipa1 protein expression levels in 122 paraffin blocks of gastric cancer and 64 normal gastric tissues were determined by substance P immunohistochemical technique. Results:The Sipa1 mRNA and Sipa1 protein levels in fresh gastric carcinoma tissues were significantly lower than those in adjacent normal tissues. Sipa1 protein was positive in 36.1%of the paraffin blocks of gastric cancer and in 73.4%of normal gastric tissues. The difference was statistically significant (P0.05). The five-year survival rates of Sipa1-negative patients were significantly lower than those of Si pa1-positive patients (P<0.01). Conclusion:Sipa1 expression is highly correlated with the biological behavior of gastric carcinoma and thus facilitates the evaluation of gastric carcinoma prognosis.

Clinical value of endoscopic metal stent placement for malignant colonic obstruction in aged people / 中华老年医学杂志

Objective To evaluate the clinica1 value of endoscopic placement of colorectal metal stents for malignant colonic obstruction in aged patients. Methods The 82 cases who were older than 65 years with malignant colonic obstruction were randomly divided into colorectal metal stenting group who were treated with endoscopic Niti-S colorectal metal stenting and emergency surgery group who were treated with tumor resection or colostomy. The average length of stay, average hospitalization charge, median survival time, one-year survival rate, perioperative mortality, and the complications such as anastomotic leakage, incision infection were analyzed. Results In colorectal metal stenting group, in 39 of 41 patients, metal stents were successfully inserted, without complications of hemorrhage, perforation and so on. And the symptoms of obstruction were effectively relieved within 2 days. The 17 cases underwent subsequent elective radical resection of colorectal carcinoma after 1 week, without complications of anastomotic leakage. The 22 cases were treated with colorectal metal stenting for palliative treatment. There were 4 cases of stent migration and 5 cases of stent obstruction again. The average length of inhospital stay was (17.9±6.7) days, the average hospitalization charge was 40 047 yuan, the median survival time was (9.4±4.4) months, and one-year survival rate was 43.9%. In emergency surgery group, 13 cases underwent tumor resection and 28 cases received colostomy. The 14 cases presented with multiple organ dysfunction, 8 cases died preoperatively, 4 cases presented with anastigmatic leakage. The average length of stay was (24.4±4.6) day, the average hospitalization charge was 61 867 yuan, the median survival time was (8.8±4.8) months, and one-year survival rate was 36.6%. Conclusions Endoscopic colorectal metal stenting is a safe, economic and effective treatment for malignant colonic obstruction in aged patients, especially reduces the risk of surgical complications, improves the aged patient's quality of life.

Changes of splenic macrophage during the process of liver cancer induced by diethylnitrosamine in rats / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>It is generally accepted that spleen plays a complex role in the tumor immunity, which would change in the different periods of cancer. In this study, we investigated the changes in the function of splenic macrophage (Mphi) in different stages of liver cancer induced by diethylnitrosamine (DEN) in rats. The aim was to support the characteristics of "two-way" and "phase" of spleen in tumor immunity.</p><p><b>METHODS</b>The model of pulmonary metastasis of liver cancer was established in forty male SD rats by DEN. In the 8th, 13th and 16th week, 10 rats were randomly chosen and sacrificed, and divided into cirrhosis, liver cancer and pulmonary metastasis groups depending on the pathological result, respectively. The other 10 rats were taken as control group. The Mphi was isolated by anchoring cultivation. The changes in ultrastructure, phagocytosis, cytokine secretion, antigen processing and presenting, and viability of splenic Mphi were detected by transmission electron microscopy, Vybrant(TM) Phagocytosis Assay, DQ(TM) Ovalbumin, and rat TNF-alpha ELISpot kits.</p><p><b>RESULTS</b>Under the electron microscope, the Mphi in the control group had some pseudopodium-like prominences, and mitochondria, ribosome, rough endoplasmic reticulum, lysosome can be found in the cytoplasm, and phagocytized RBC. In the liver cirrhosis and liver cancer group, Mphi had more prominences, meanwhile much more mitochondria, ribosome, rough endoplasmic reticulum, lysosome can be found in the cytoplasm, especially in the liver cancer group. In the pulmonary metastasis group, the Mphi was swelling, with few organelle. As compared to the control group, the function of splenic Mphi increased in cirrhosis and cancer groups, but decreased in metastasis group (phagocytosis rate: (84.7 +/- 1.9)%, (89.5 +/- 3.1)%, and (36.0 +/- 2.6)% vs (75.6 +/- 1.7)%, P < 0.05, P < 0.01; viability: (1.53 +/- 0.15)%, (1. +/- 0.14)%, and (1.12 +/- 0.29)% vs (1.48 +/- 0.17)%, P < 0.05, P < 0.01; TNF-alpha secretion: (741.0 +/- 52.9)%, (1126.2 +/- 174.5)%, and (313.8 +/- 50.8)% vs (626.6 +/- 24.6)%, P < 0.05, P < 0.01; positive cell rate of antigen processing and presenting: (24.03 +/- 1.87)%, (27.95 +/- 2.63)%, and (10.46 +/- 2.16)% vs (16.45 +/- 1.86)%, P < 0.01).</p><p><b>CONCLUSIONS</b>In the stage of cirrhosis and early cancer, the immune functions of splenic Mphi were reinforced. It may promote the non-specificity tumor immunity. On opposite, in the stage of pulmonary metastasis, the immune functions of splenic Mphi were impaired. It may lead to the decrease of tumor immunity.</p>

Morphological changes of blood spleen barrier in portal hypertensive spleen / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>The pathogenesis of hypersplenism and the immune function of the spleen in patients with portal hypertension (PH) remain obscure. This study aimed to evaluate the morphological changes of blood spleen barrier in spleen with hypersplenism due to PH and provide evidence for an in-depth investigation of the immune function of the spleen with hypersplenism and the mechanism of hypersplenism.</p><p><b>METHODS</b>Spleen samples from 12 portal hypertensive patients and 4 patients with traumatic ruptures of spleen were examined. The samples of spleen were made into pathological sections, stained with Masson trichrome stain, Gomori stain, and CD68, CD34 immunohistochemistry, and were examined microscopically for the changes in the distribution of collagen fibers, reticular fibers, macrophages, and vascular endothelial cells. The changes in ultrastructure of macrophages and endothelial cells in marginal zone were also evaluated by transmission electron microscopy.</p><p><b>RESULTS</b>As compared to the normal spleen, the density of macrophage in the PH spleen was decreased, but the macrophages were mainly located in the marginal zone and distributed around the splenic corpuscle, with many villi and pseudopodium-like protrusion on the cell surface. The accrementition of collagen fibers was obvious around the splenic corpuscle and central artery. The increased reticulate fibers encircled the splenic corpuscle with more connection between the fibers. The vascular endothelial cells were in diffused distribution, without any regionality in PH spleen, but the vessel with enlarged lumina increased in red pulp.</p><p><b>CONCLUSIONS</b>The morphological changes of the blood spleen barrier can be one of the pathological fundaments for the abnormality of the immune function and the increased destruction of blood cells located in the spleens of patients with PH. However, this still entails clarification.</p>