SMARCA2 deficiency in NSCLC: a clinicopathologic and immunohistochemical analysis of a large series from a single institution

BACKGROUND@#SMARCA2 (SWI/SNF Related, Matrix Associated, Actin Dependent Regulator of Chromatin, Subfamily A, Member 2) is an important ATPase catalytic subunit in the switch-sucrose nonfermenting (SWI/SNF) complex. However, its relationship with the pathological features of NSCLC and its prognosis remain unclear.@*METHODS@#We retrospectively reviewed 2390 patients with surgically resected NSCLC, constructed tissue microarrays (TMAs) and performed immunohistochemical assays. We analyzed the correlation of SAMRCA2 with clinicopathological features and evaluated its prognostic value.@*RESULTS@#Among 2390 NSCLC cases, the negative expression ratios of SAMRCA2, SMARCA4, ARID1A, ARID1B and INI1 were 9.3%, 1.8%, 1.2%, 0.4% and 0%, respectively. In NSCLC, male sex, T3 and T4 stage, moderate and poor differentiation, tumor ≥ 2 cm, Ki67 ≥ 15%, SOX-2 negative expression, middle lobe lesion and adenocarcinoma were relative risk factors affecting SMARCA2-negative expression. In lung adenocarcinomas, high-grade nuclei, histological morphology of acinar and papillary, solid and micropapillary and TTF-1-negative expression were relative risk factors affecting SMARCA2-negative expression. Kaplan-Meier survival analysis showed that the OS was shorter in the SMARCA2-negative group. Multivariate survival analysis revealed that SMARCA2-negative expression was an independent factor correlated with a poor prognosis in NSCLC.@*CONCLUSION@#In conclusion, SMARCA2-negative expression is an independent predictor of a poor outcome of NSCLC and is a potential target for NSCLC treatment.

Current status and perspectives of integrated traditional Chinese and Western medicine therapy for pancreatitis / 临床肝胆病杂志

Integrated traditional Chinese and Western medicine therapy for pancreatitis has been used since the 1960s.According to the clinical manifestations and traditional Chinese medicine (TCM) syndrome differentiation of pancreatitis,pancreatitis was named true heart pain with cold limbs caused by spleen disease.Syndrome differentiation of acute pancreatitis and treatment with modified Dachaihu decoction achieved good clinical effects.After the 1990s,the research focus of pancreatitis research was shifted to severe acute pancreatitis (SAP).The clinical course of SAP was divided into three phases,and different therapeutic regimens were given.Clinical studies achieved good therapeutic effects in terms of cure rate and fatality rate.After 2000,integrated traditional Chinese and Western medicine therapy for chronic pancreatitis has been promoted systemically,and the cooperation between traditional Chinese medicine,endoscopic techniques,and operative treatment helps to significantly improve pain control,nutritional status,and incidence of complications.

A study on alleviation of acute lung injury in rats with bowel repletion pattern by mesenteric lymph drainage and its mechanism / 中国中西医结合急救杂志

Objective To observe the effects of mesenteric lymph drainage on acute lung injury and expression of p38 mitogen activated protein kinase (p38MAPK) signal pathway in rats with bowel repletion pattern. Methods Thirty male Wistar rats were randomly divided into three groups according to random number table method, namely sham operation group (sham group), bowel repletion model group (model group) and mesenteric lymph drainage group (drainage group), 10 rats in each group. The rat model of bowel repletion was established by ischemia/reperfusion (I/R) method, firstly 1 hour occlusion of superior mesenteric artery (SMA) to induce ischemia followed by reperfusion for 2 hours. In the rats of drainage group, the drainage of mesenteric lymph duct began at the end of model establishment and persisted for 3 hours. In the rats of sham group, the SMA and mesenteric lymph ducts were exposed with blunt dissection, and then they were immediately placed back into the abdominal cavity. After 3 hours of mesenteric lymph drainage, the lung and ileum tissues of rats in each group were harvested for evaluation of pathohistological changes and for the determination and comparison of myeloperoxidase (MPO) activity changes; the levels of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) in bronchoalveolar lavage fluid (BALF) were detected by enzyme linked immunosorbent assay (ELISA), and the expressions of Toll-like receptor 4 (TLR4) mRNA and p38MAPK mRNA in the lung tissues were measured by fluorescence quantitative polymerase chain reaction (PCR).Results Under the light microscope, the pulmonary capillaries markedly dilated and congested, the interstitium width of lung increased with a large amount of inflammatory cells infiltration, the intestinal mucosal layer becoming thinner with detachment of intestinal villi and a large amount of inflammatory cells infiltration were detected in rats of model group. Compared with those in sham group, the levels of TNF-α and IL-6 in BALF, the MPO activity of lung and ileum tissues, and the expressions of TLR4 mRNA and p38MAPK mRNA in the lung tissues were significantly increased in model group.Compared with those in model group, the pathohistological damages in lung and ileum tissues were ameliorated, the levels of TNF-α and IL-6 in BALF, the MPO activity of lung and intestinal tissues and the expressions of TLR4 mRNA and p38MAPK mRNA in the lung tissues were lower in the rats of drainage group [TNF-α in BALF (ng/L): 858.55±27.16 vs. 1 680.58±105.62; IL-6 in BALF (ng/L): 0 vs. 484.71±5.43; MPO activity of lung (U/g): 0.95±0.13 vs. 1.36±0.11; MPO activity of ileum tissues (U/g): 0.75±0.13 vs. 1.30±0.16; TLR4 mRNA: 0.21±0.11 vs. 0.69±0.13, p38MAPK mRNA: 0.21±0.13 vs. 0.47±0.09; allP < 0.05].Conclusion Mesenteric lymph drainage can alleviate acute lung injury in rats with bowel repletion, and its mechanism may be related to the reduction of the expressions of TLR4 mRNA and p38MAPK mRNA and the release of TNF-α and IL-6 in lung tissues.

Effects of macrophages and monocyte chemoattractant protein-1 during experimental choroidal neovascularization / 中华实验眼科杂志

Background Choroidal neovascularization (CNV) is one of the primary causes leading to visual damage in many fundus diseases.Many evidences indicate that macrophage activation and monocyte chemoattractant protein-1 (MCP-1) play important roles in CNV.However, the dynamic expression of macrophage and MCP-1 in the initial stage of CNV is not clear.Objective This study was to investigate the dynamic changes of F4/80 and MCP-1 expressions in retina-choroid tissue with experimental CNV.Methods Laser-induced CNV models were monocularly established in 105 SPF 8-week-old male wild type C57BL/6 mice.The mice were sacrificed at 6,12,24, 48 and 72 hours after photocoagulation, respectively, and the retina-choroid tissue sections and choroidal flatmounts were prepared.The histopathological examination was carried out to observe the changes of morphology and structure as well as inflammatory response in CNV.The expression and distribution of F4/80 and MCP-1 protein in retinachoroid were detected by double immunofluorescence technique.The expression and distribution of F4/80 in choroid were examined by immunofluorescence.The relative expression levels of F4/80 mRNA and the content of MCP-1 protein in RPE-choroid complex were assayed using real-time quantitative PCR and ELISA,respectively.The use and care of the mice complied with the Regulation for the Administration of Affair Concerning Experimental Animals by Ethic Committee of Experimental Animals of Nanjing Medical University.Results The rupture of Bruch membrane, RPE, outer nuclear layer and choroid was exhibited under the optical microscope 6 hours after photocoagulation.Infiltration of inflammatory cells and tissue edema were seen as the lapse of photocoagulation time, and proliferation of vascular endothelial cells was found 72 hours after photocoagulation.F4/80 was expressed in photocoagulation area 6 hours later, and MCP-1 was expressed around the area.With the lapse of photocoagulation time,the expression intensity of MCP-1 weakened and that of F4/80 enhanced.The contents of MCP-1 protein in RPE-choroid complex were (31.25±4.73), (276.31 ±4.20), (331.95 ±5.86), (221.24±4.42), (179.89 ± 4.10) and (130.80 ± 5.90) pg/mg in the normal control group, photocoagulation 6-, 12-, 24-, 48-and 72-hour groups, respectively,with a significant difference among the groups (F=1 416.46 ,P＜0.01).The contents of MCP-1 protein peaked at 12 hours after photocoagulation and then gradually declined.The expression levels of MCP-1 protein in different time groups were higher than those in the normal control group (all at P＜0.01).A significant difference in F4/80 mRNA expression in RPE-choroid complex was also found among the groups (F =762.72, P＜0.01, and a gradually raising tendency was seen over time, showing evidently increase in comparison with the normal control group (all at P＜0.01).Conclusions Inflammatory response occurs in the early stage of experimental CNV.MCP-1 responds to the CNV at early stage,and the accumulation and activation of macrophage play an important role in the development of CNV.

Analysis of components of mesenteric lymph in rats with severe intraperitoneal infection / 中华危重病急救医学

Objective To investigate the components of mesenteric lymph of the rats with severe intraperitoneal infection,and inquire into the effect of intestinal lymphatic pathway in severe intraperitoneal infection.Methods Twenty-four male Wistar rats were randomly divided into two groups according to random number table method,namely model group and sham group with 12 rats in each group.The rat model of severe intraperitoneal infection was reproduced by injecting artificial gastric juice and E.coli intraperitoneally.Mesenteric lymph in both groups was collected 4 hours after the reproduction of the model,and white blood cells were counted and classified.The levels of endotoxin,alkaline phosphatase (AKP),lactate dehydrogenase (LDH),creatine kinase (CK),glutamine transferase (GST),protein and cytokines of mesenteric lymph were determined.Results Compared with the sham group,there was an increase in the neutrophil ratio in mesenteric lymph (0.167 ± 0.004 vs.0.610 ± 0.006,t=33.520,P＜0.001),however the percentage of both macrophages (0.009 ± 0.001 vs.0.020 ± 0.004,t=-6.677,P＜0.001) and lymphocytes (0.824 ± 0.005 vs.0.921 ± 0.004,t=-31.471,P＜0.001) was decreased in model group.Compared with sham group,the levels ofendotoxin (kEU/L:0.346 ±0.022 vs.0.186 ±0.001,t=18.103,P＜0.001),AKP [U (king unit):13.97 ± 5.55 vs.3.76 ± 0.18,t=4.503,P=0.006],LDH (U/L:2 827.45 ± 1 940.32 vs.712.68 ± 14.09,t=2.670,P=0.044),CK (kU/L:2.19 ± 1.21 vs.0.70 ± 0.01,t=3.035,P=0.029),GST (kU/L:12.33 ± 6.53 vs.1.36 ± 0.39,t=4.105,P=0.009) were all significantly elevated.The concentration of protein (g/L:4.40 ± 0.48 vs.2.84 ± 0.16,t=6.882,P=0.001),tumor necrosis factor-α [TNF-α (ng/L):499.39 ±76.36 vs.180.90 ± 70.98,t=7.483,P＜0.001],interleukin-6 [IL-6 (μg/L):13.74 ± 0.78 vs.-0.07 ± 0.07,t=52.972,P＜0.001],intercellular adhesion molecule-1 [ICAM-1 (ng/L):2 754.19 ±221.48 vs.1 362.85 ±393.43,t=6.891,P＜0.001] and monocyte chemo-attractant protein-1 [MCP-1 (μg/L):28.23 ± 1.77 vs.24.87 ± 1.15,t=3.561,P=0.007] and high mobility group protein-1 [HMGB-1 (ng/L):1 392.78 ± 572.42 vs.564.17 ± 21.32,t=3.543,P=0.016] in mesenteric lymph in model group were significantly higher than those in sham group.Conclusion Intestinal lymphatic pathway maybe the early pathway for the production of remote organ injury caused by severe intraperitoneal infection.

The effect of Qingfei Chengqi decoction on lung tissue cell apoptosis in rats with severe intraperitoneal infection / 中国中西医结合急救杂志

Objective To observe the effects of Qingfei Chengqi decoction on lung tissue cell apoptosis and its associated protein,tumor necrosis factor-α(TNF-α),interleukin-6(IL-6)of lung tissues in rats with severe intra-peritoneal infection(SII). Methods Thirty male Wistar rats were randomly divided into three groups:sham operation group,model group and traditional Chinese medicine(TCM)group(each n=10). Simulating clinical pathophysiological process of digestive tract perforation,the rat model of SII was reproduced by injecting E. coli intraperitoneally. The TCM group was treated by gavage with Qingfei Chengqi decoction one day in advance of the study,3 times per day,once 2 mL. The same amount of nutrient broth which contained 10%barium sulfate(BaSO4) to replace the bacteria solution was injected into the sham operation group. Six hours after model establishment,all rats were killed and lung tissues were harvested for pathohistological evaluation and for the determination of apoptosis rate with TdT-mediated dUTP nick-end labeling(TUNEL)method,of Bax,Bcl-2 protein expression with Western Blot method,and of the level of TNF-α,IL-6 with enzyme-linked immunosorbent assay(ELISA)method,and the pathological changes of lung tissues were observed under the light and electron microscopes. Results Compared with sham operation group,the apoptosis rate〔(12.7±5.4)%vs. 0〕,the expression of Bax protein〔absorbance(A)value:8 416.89±875.16 vs. 6 654.09±1 130.18〕,the level of TNF-α(ng/L:3 132.56±457.96 vs. 1 948.55±269.32), the level of IL-6(ng/L:75.14±1.63 vs. 31.17±0.81)of lung tissues were significantly increased(all P<0.05), meanwhile Bcl-2 protein expression decreased observably(A value:7 490.59±200.34 vs. 12 289.02±535.93,P<0.05)in model group induced by SII. After treatment with TCM,apoptosis rate〔(7.9±0.3)%〕,the expression of Bax protein(A value:7 619.50±999.30),the level of TNF-α(ng/L:3 114.34±454.32)and IL-6(ng/L:52.46±0.96) of lung tissues were decreased and Bcl-2 protein expression(A value:11 155.07±1 018.87)increased(all P<0.05) compared with model group. General observation:the color of lung tissues was uniform in sham operation group;the lung tissues of model group swelled obviously,and parts of lung tissues had patches of ecchymosis and hemorrhage. The light microscope showed:the pulmonary vessels,the alveolar interstitium,alveolar epithelium and lobular interstitium of sham operation group were all normal,while in the model group,the pulmonary interstitium was edematous and hemorrhagic,and in the alveolar cavities there was infiltration of inflammatory cells. Under the electron microscope, the lung tissue type Ⅱ alveolar epithelial cells of model group were increased,and they had morphological changes in various degrees,such as cell shrinkage and change becoming round,and cell nucleus presenting irregular in shape. After treatment with TCM,the above pathological changes were all alleviated compared with those in the model group. Conclusions Qingfei Chengqi decoction can ameliorate the SII leading to acute lung injury,and reduce the cell apoptosis rate of lung tissues in SII rats. Its mechanism may be related to the intervention of above TCM which can lower the levels of inflammatory media and elevate the protein expression of Bcl-2.

A study on the change of matrix metalloproteinase-2 and peripheral white blood cells of acute cerebral in-farction patients at different altitudes / 中国神经精神疾病杂志

Objective To investigate the changes of the serum levels of matrix metalloproteinase- 2 (MMP-2) and peripheral blood leukocytes content and its relationship with the severity of cerebral infarction in acute cerebral in-farction(ACI)patients at different altitudes (high, middle and low). Methods One hundred thirty-nine cases and 150 healthy controls were included in the present study. Enzyme linked immunosorbent method was used to detect MMP-2 and WBC levels. Results MMP-2 levels increased as the altitude increased in controls. The MMP-2 in a descending or-der was 1.41±0.39 in Haixi (high altitude), 1.37±0.27 in Xining (middle altitude) and 1.28±0.21 in Sichuan (low altitude) (P<0.05). The serum levels of MMP-2 were significantly increased at 7 d at different altitudes (5.75±1.19, 5.23±1.12 and 4.15 ± 0.97 in low, middle and high altitudes, respectively). The WBC were significantly increased at different alti-tudes (12.93±2.11, 12.11±1.74 and 11.15±1.68 in low, middle and high altitudes, respectively) within 48 h in severe ACI group (P<0.05). MMP-2 levels in different altitudes were positively associated with the infarction size and the degree of neurological deficit, while were negatively correlated with the prognosis. The WBC in large infarction group were positive-ly correlated with the infarct size. Conclusions The levels of MMP-2 and WBC in different altitudes may be helpful in determining the ACI lesion size and the severity of the illness as well as estimating the prognosis.

Effect of mesenteric lymph drainage on intestinal barrier function in rats with severe intraperitoneal infection / 中华胃肠外科杂志

<p><b>OBJECTIVE</b>To investigate the effect of mesenteric lymph drainage on intestinal barrier function in severe intraperitoneal infection (SII).</p><p><b>METHODS</b>Thirty healthy male Wistar rats were randomly divided into 3 groups(model group, drainage group and control group). SII model rats were prepared by injecting E.coli intraperitoneally. Rats in drainage group rats underwent mesentery lymphatic duct ligation and drainage 2 hours after model induction, and those in control group received equal amount of 10% BaSO4 nutrient broth injection intraperitonerally. Six hours after model induction, rats were sacrificed. The intestinal samples were collected for pathology analysis and content of DAO and concentration of TNF-α, IL-6. Content of D-lactate in blood plasma was detected.</p><p><b>RESULTS</b>Under light microscopy, ileum mucosa tissue structure of model group was disordered. Under transmission electron microscopy, intestinal mucosal epithelial cells of model group swelled obviously, close connection was destructed, and early apoptosis cells occurred. After mesentery lymph drainage, intestinal mucosa tissue structure was improved obviously, endoplasmic reticulum and mitochondria of epithelium swelled mildly. The contents of intestinal tissue DAO in drainage group, model group and control group were (5.9±0.4) U/L, (3.0±0.1) U/L and (18.3±2.1) U/L respectively. There was significant difference among groups (P<0.05). Compared with control group [(45.4±37.9) μg/L], the plasma content of D-lactate in model group [(256.0±177.2) μg/L] increased significantly (P<0.05). The plasma content of D-lactate in drainage group [(136.9±21.5) μg/L] was not significantly different compared with model group (P>0.05), but was significantly higher compared to control group (P<0.05). Compared with control group, model group had significantly higher levels of TNF-α [(3431.3±23.9) ng/L vs. (2730.0±408.7) ng/L] and IL-6 [(86.3±1.6) ng/L vs. (30.2±0.9) ng/L] (P<0.05), while the TNF-α was (2653.2±324.1) ng/L, and the IL-6 was (50.9±0.7) ng/L in drainage group, which were significantly lower compared with model group (P<0.05).</p><p><b>CONCLUSION</b>Mesenteric lymph drainage can obviously improve intestinal barrier function in severe intraperitoneal infection and may play a protective role in intestinal mucosa.</p>

The mechanism of miR-181c induced neuroprotection by hypoxia preconditioning in rats / 中华神经科杂志

Objective To investigate the neuroprotective effect of miR-181c on hypoxia-preconditioned ischemia in rats and its mechanism.Methods Thirty-nine male SD rats were randomly divided into 5 groups of control group,sham-operated group,middle cerebral artery occlusion (MCAO)group,hypoxia-preconditioned group,hypoxia-preconditioned and MCAO group.Infarct volume and behavioral deficits were quantified.Real-time PCR was applied to detect the expression levels of miR-181c and Western blotting was used to verify the target protein of mt-cox1.Results Under the treatment of hypoxia-preconditioned,the neurological impairment was alleviated and the infarct volume was reduced significantly from 22.50％ ±2.96％ to 16.40％ ±3.13 ％ (t =5.26,P ＜0.01).The expression of miR-181c was decreased significantly in hypoxia-preconditioned and MCAO group than that in MCAO group (1.89 ± 0.14 vs 3.05 ± 0.26,t =6.10,P ＜ 0.01),and the expression of mt-cox1 protein was also significantly decreased (0.54 ± 0.07 vs 0.93 ± 0.04,t =8.01,P ＜ 0.01).Conclusion Hypoxia-preconditioned may attenuate the ischemic injury in SD rats,which may be related to the down-regulation of the expression of miR-181c,therefore increasing the expression of its targeted protein mt-cox1.