Objective To investigate the change in blood lymphocyte levels and lymphocyte percentage in critical patients,including those with non-infectious systemic inflammatory response syndrome (SIRS),sepsis and severe sepsis.Methods Clinical data of 423 patients admitted to intensive care unit (ICU) of Department of Emergency of Peking University Third Hospital from January 2011 to September 2013 were retrospectively analyzed.Among these patients,there were 54 with non-infectious SIRS,177 with sepsis and 192 with severe sepsis.150 of them died,and 273 survived.The white blood count (WBC),neutrophil (N),lymphocyte count,lymphocyte percentage,lactic acid,serum high-sensitivity C-reactive protein (hs-CRP) and procalcitonin (PCT) were determined at admission,and acute physiology and chronic health evaluation Ⅱ (APACHE Ⅱ) was estimated.The patients were divided into different groups according to diagnosis and prognosis to compare the value of the indexes,and the correlation between lymphocyte count and other markers were analyzed by Spearman relative analysis.Results The older the patients,the more severe the disease.In the non-infective SIRS,sepsis and severe sepsis groups,APACHE Ⅱ scores (7.78 ± 3.72,13.85 ± 7.22,24.00 ± 9.79),time of stay in hospital [days:6.0 (1.0,9.0),12.0 (8.0,22.0),19.5 (7.0,29.0)],the mortality rate (0,10.2％,52.6％),WBC (× 109/L:7.59 ± 3.27,8.94 ± 3.95,10.32 ± 5.50),N (0.685 ± 0.132,0.778 ± 0.135,0.831 ±0.086),hs-CRP [mg/L:4.60 (2.80,7.52),23.58 (13.49,49.22),59.77 (19.36,110.62)] andPCT [μg/L:0.05 (0.05,0.05),0.09 (0.05,0.61),0.63 (0.10,5.25)] showed gradually increasing tendency (all P=0.000).Serum lymphocyte count levels [× 109/L:1.53 (0.89,1.88),0.90 (0.65,1.42) and 0.80 (0.50,1.12)],lymphocyte percentage (0.225 ± 0.122,0.138 ± 0.097,0.106 ± 0.070) showed gradually decreasing tendency (P＜0.05 or P＜0.01).There was significant difference in blood lactic acid among three groups [blood lactic acid were 2.40 (1.30,5.10),1.10 (0.80,2.00) and 1.40 (1.00,2.50) mmol/L in the non-infective SIRS,sepsis and severe sepsis groups respectively,P=0.000].Age (years old:76.71 ± 12.21 vs.73.21 ± 14.49),APACHE Ⅱ score (24.69 ± 9.58 vs.13.91 ± 8.41),time of stay in hospital [days:12.0 (4.0,28.0) vs.11.0 (8.0,22.0)],WBC (× 109/L:10.29 ± 5.82 vs.8.89 ± 3.98),N (0.809 ± 0.130 vs.0.776 ± 0.120),lactic acid [mmol/L:1.80 (1.10,2.90) vs.1.30 (0.90,2.49)],hs-CRP [mg/L:50.94 (19.21,97.13) vs.21.71 (6.39,54.40)] and PCT [μg/L:0.74 (0.13,5.83) vs.0.08 (0.05,0.59)] levels in the death group were higher than those in the survival group (P＜0.05 or P＜0.01),serum lymphocyte count levels [× 109/L:0.90 (0.50,1.29) vs.1.05 (0.70,1.54)],lymphocyte percentage (0.123 ± 0.098 vs.0.143 ± 0.097) level in the death group were obviously lower than those in the survival group (P＜0.01 and P＜0.05).Serum lymphocyte count levels were negative correlated with N (r=-0.597,P=0.000),hs-CRP (r=-0.298,P=0.000),PCT (r=-0.304,P=0.000),APACHE Ⅱ (r=-0.124,P=0.000),and positively correlated with lymphocyte percentage (r=0.691,P=0.000),and non correlations was found with WBC (r=0.082,P=0.091) and lactic acid (r=0.073,P=0.132).Conclusion The serum lymphocyte levels in the critically ill patients are related with the severity of sepsis,and monitoring the change in lymphocyte may be an indicator for evaluating the illness and effect of treatment.

Objective To observe the expressions of RANTES , FKN and IP-10 at mRNA and pro-tein levels in human lung epithelial cells (A549) infected by respiratory syncytial virus (RSV), and to eval-uate the changes of them interfered with 15-deoxy-delta12,14prostaglandin J2(15d-PGJ2), rosiglitazone or 2-chloro-5-nitrobenzanilide ( GW9662 ) .Methods A549 cells were seeded in 6-well culture plates and cul-tured overnight in F12K culture solution.Then they were randomly divided into five groups , including group A (15d-PGJ2+RSV group), group B (rosiglitazone+RSV group), group C (DMSO+RSV group), group D (GW9662+rosiglitazone+RSV group) and group E (cell control group).Cells and supernatants were harves-ted from each group at different time points (12 h, 24 h and 48 h) of culture.The expressions of RANTES , FKN and IP-10 at mRNA and protein levels were measured by ELISA and real-time quantitative RT-PCR analysis.Results The expressions of RANTES , FKN and IP-10 at mRNA and protein levels in group C were significantly higher than those in group E at time points of 12 h, 24 h and 48 h (all P<0.05).In group C, the expressions of the three chemokines at mRNA level reached a peak at 24 h, but began to de-crease at 48 h, which showed no statistical significance compared with those at 12 h (all P>0.05).Moreo-ver, the expressions at protein level were peaked at 48h, and had significant difference with those expressed at 12 h and 24 h (all P<0.05).Compared with group C, the expressions of the three chemokines both at mRNA level and protein level were decreased in group A and B as the dose was increased (all P<0.05), and the lowest levels were observed with the intervention of 20 μmol/L of 15d-PGJ2 in group A and 30μmol/L of rosiglitazone in group B .Conclusion The expressions of RANTES , FKN and IP-10 at mRNA and protein levels were increased with RSV infection , and the peaks of mRNA level and protein level were respectively achieved at 24 h and 48 h after infection.PPARγagonists played an anti-inflammatory role through inhibiting the expressions of the three chemokines both at mRNA level and protein level in a dose -de-pendent manner .

Objective To investigate the expression of TIP 30 protein in gastric cancer tissues,and effect of TIP30 over-expression on migration and invasion of gastric cancer cell line SGC7901.Methods Immunohistochemistry streptavid-in-peroxidase (SP) methods were used to detect the expression levels of TIP30 in 93 cases of gastric cancer tissues.Previously constructed pcDNA3.1-TIP30 plasmid were transiently transfected into SGC7901 cells.The proliferation of cells were detected by using MTT assay when TIP30 was overexpressed.Transwell assay to determine migration and invasion ability of SGC7901.Western blot was used to examine the changes of concentration of E-cadherin,N-cadherin and MMP9.Results The positive expression rate of TIP30 was 39％ significantly lower in gastric cancer tissues than 92％ in normal gastric mucosa tissues (x2 =32.68,P ＜ 0.05),there was a significant correlation between reduced expression of TIP30 and depth of infiltration,including nodal metastasis,TNM stage (x2 =3.535,7.421,6.754,all P ＜ 0.05);MTT showed that the proliferation of SGC7901 cells in the pcDNA3.1-TIP30 transfected group significantly decreased when TIP30 was overexpressed at respective time of 72,96 hours (t =6.528,7.249,both P ＜ 0.05),Transwell assay showed that overexpression of TIP30 significantly decreased migratory and invasive numbers of SGC7901 cells (t =5.769,P ＜ 0.05;t =7.886,P ＜ 0.05);the expression level of MMP-9 and N-cadherin in TIP30 overexpressing cells group significantly decreased (t =9.811,10.362,both P ＜ 0.05),mean while E-cadherin expression was significantly higher than before (t =6.137,P ＜ 0.05).Conclusion TIP30 protein is low expressed in gastric cancer and the overexpression of TIP30 inhibits the proliferation,migration and invasion of gastric cell line SGC7901.

Objective: To establish a method for repairing extremities with extensively deep burn using large piece of fresh allogeneic scalp spliced by Meek glue combined with autologous microskin and observe its effect. Methods: Medical records of two male patients with extremely extensive deep burn admitted to our hospital from May to November in 2018 were retrospectively analyzed. Two patients aged 44 and 25 years respectively, with total burn area of 90% and 97% total body surface area (TBSA) and full-thickness burn area of 85% and 70% TBSA, respectively. Preoperatively, the surgical area on the extremities was calculated to estimate the necessary amount of allogeneic scalp and Meek miniature skin. The large piece of fresh allogeneic scalp spliced by Meek glue combined with autologous microskin was prepared according to the methods described as follows. Thin medium-thickness fresh scalps with 3% TBSA and 0.30-0.35 mm in depth were harvested from each donor and spliced into a large piece with epidermis upward by spraying Meek glue. Then the spliced scalp was punched after covered with a single-layer gauze. Autologous microskin was transported onto the dermis of fresh large piece of allogeneic scalp by traditional floating method. Bilateral extremities with full-thickness burn of two patients were selected for self-control. The left upper extremity was denoted as treatment group while the right upper extremity was denoted as control group in Patient 1. The right lower extremity was denoted as treatment group while the left lower extremity was denoted as control group in Patient 2. Wounds in the treatment group were treated with fresh large piece of allogeneic scalp spliced by Meek glue and autologous microskin with expansion ratio of 1∶15 after escharectomy, while wounds in control group received grafting of Meek miniature skin with expansion ratio of 1∶6 and or 1∶9 after escharectomy. The donors of allogeneic scalp were 32 males who were the relatives or friends of the patients, aged 21-50 years, with scalp area of (548±48) cm². The healing conditions of donor sites of scalp were observed on post operation day 10, and were followed up within 3 months after operation to observe whether forming alopecia and hypertrophic scar or not. Wound healing condition was evaluated during follow-up in post operation week (POW) 2-5 and 4 months after operation. Wound coverage rates were calculated in both treatment and control groups in POW 2, 3, 4, and 5. Results: The donor sites of all allogeneic scalp of donors healed completely on post operation day 10. There was no alopecia or hypertrophic scar within 3 months after operation for follow-up. In POW 2, allogeneic scalp grafts basically survived in treatment group without obvious exudation, and most of the Meek miniature skin survived in control group with obvious exudation. Part of allogeneic scalp grafts dissolved and detached in treatment group in POW 3, and the surviving grafts scabbed. The eschar detached and new epithelium was observed in treatment group in POW 4 and 5. In POW 3-5, surviving Meek miniature skin in control group creeped and was incorporated, and the wounds shrank. Hypertrophic scar was observed in both treatment and control groups 4 months after operation, without obvious difference in scar as a whole. The wound coverage rates were respectively 84%-98% and 76%-92% in treatment group of two patients in POW 2-5, close to or higher than those of control group (35%-97% and 28%-81%, respectively). Conclusions The study establishes a novel method for splicing fresh allogeneic scalps into a large piece as the covering of microskin, which has good effect for repairing extensively deep burn wounds. Considering that allogeneic skin is scarce, this method may be a new option in clinical treatment for extensively deep burn patients.

Objective:To explore the risk factors of lymph node metastasis (LNM) in early gastric cancer (ECG), and establish a risk-prediction model based on LNM.Method:Four hundred and twenty-seven EGC patients undergoing curative radical gastrectomy were enrolled in this study. The risk factors for LNM of ECG were analyzed with Logistic regression. LNM risk was stratified and risk-predicting model was established. The risk-predicting model was measured by area under ROC curve. According to the same standard, clinical data of 133 patients with EGC who underwent radical surgery were selected for external verification of the model.Results:The frequency of LNM was 13.3% (32/427) in EGC patients. The LNM ratio of intramucosal carcinoma and submucosal carcinoma was 1.3% (3/237), 15.3% (29/190) respectively. Ulcer presence, tumor size >2 cm, undifferentiated tumor, submucosal invasion, neural invasion, and vascular tumor thrombus were significantly associated with LNM in EGC patients ( χ2=3.408, 16.379, 4.808, 29.804, 25.305, 47.120, respectively P<0.05). Multivariate analysis suggested that ulcer presence, tumor size >2 cm, depth of invasion, neural invasion, and vascular tumor thrombus were independent predictors of LNM in EGC patients, ( OR=0.326, 2.924, 11.824, 13.047, 7.756, respectively P<0.05). LNM predicting model is established, P=e^x/(1+e^x),x=-4.792-1.122 ulcer presence+1.073 tumor size+2.470 depth of invasion+2.569 neural invasion+2.048 vascular tumor thrombus,ROC-AUC of risk-predicting model was 0.845, the best cut-off was 0.094, the sensitivity was 72.70%, the specificity was 77.20%. The external verification result revealed the AUC of ROC was 0.840. The four-grid table is constructed by predicting model results and the postoperative pathological examination. The sensitivity and specificity of the model are calculated to be 82.35% and 68.96%, respectively. Conclusions:EGC patients with ulcer presence, tumor size >2 cm, depth of invasion, vascular tumor thrombus, and neural invasion have higher risk of LNM, the risk-predicting model can identify the high probability of LNM .

Objective:To investigate the expression of KLF11 in gastric cancer tissues and cell lines as well as its effect on proliferation and apoptosis of human gastric cancer cells BGC823.Methods:Sixty pairs gastric cancer tissues and corresponding adjacent tissues were collected. The expression of KLF11 mRNA in gastric cancer tissues and their adjacent tissues was detected by RT-PCR. The expression of KLF11 was detected in gastric cancer cells. KLF11 expression was silenced. The proliferation of cells were detected by using MTT assay. Flow cytometry was used to detect the cell cycle and cell apoptosis rate. Western bloting was used to examine the changes of concentration of proteins associated with cell cycle,cell apoptosis and Wnt/β-catenin signaling pathway related proteins. The activity of Caspase3 enzyme was detected by spectrophotometry.Results:The relative expression of KLF11 mRNA in gastric carcinoma tissues was significantly higher than that of the adjacent tissues ( t=11.38, P<0.05). Its expression was related to tumor size, depth of invasion, lymph node metastasis and TNM clinical stage (all P<0.05). The proliferation of BGC823 cells was significantly suppressed after KLF11 silencing ( F=19.56, P<0.05), and the cell cycle was arrested in G 0/G 1 phase [(41.40%±0.98%) vs. (66.53%±1.01%), F=32.69, P<0.05]. Meanwhile, the apoptosis rate was significantly increased by KFL11 silencing [(41.44%±1.59%) vs. (15.42%±0.86%), F=35.35, P<0.05]. The results of Western blotting revealed that the expression of Bax and Cleaved Caspase3 was significantly increased ( F=23.33, 33.63; both P<0.05), wheras that of β-catenin, Bcl-2, CyclinD1and CyclinE was significantly reduced ( F=22.21, 16.24, 26.75, 33.42; all P<0.05). The activity of Caspase3 enzyme was enhanced ( F=16.56, P<0.05). Conclusion:KLF11 was highly-expressed in gastric cancer tissues and cells, KFL11 silencing could inhibit gastric cancer cells proliferation and induce cell apoptosis via Wnt/β-catenin signaling pathway.

As national strategic resources,traditional Chinese medicine (TCM) resource is the material basis for the pharmaceutical industry and health services.The high-efficiency utilization of TCM resource is a major strategic issue that realize the resource conservation and environment friendly recycling economy,guarantee the sustainable development of medicine.But currently,on one hand,the wild TCM resource was seriously damaged and in a serious shortage of stock,the cultivation variety was degenerated,the tending of endangered medicinal materials and the development of alternative varieties were faced many problems.On the other hand,the limited resources cannot be effectively utilized,which results in the waste of resources.Therefore,two ecological restricted resources,poria cocos and Chinese gall,were studied for comprehensive utilization of resources to explore the comprehensive exploitation and utilization of TCM resource as well as the pattern of sustainable development.We suggested that TCM resource should be taken into account as a whole for comprehensive utilization.We should especially pay attention to basic research for the sustainable development of TCM resource,discovery of bioactive substance,excavation and transformation for functional components,the use of biotechnology,the product development,the formation and extension of industry chain.So as the comprehensive exploitation and utilization of TCM resources and sustainable development would be realized.

OBJECTIVEThe purpose of this study was to investigate the effect and mechanism of Vav3 gene on the proliferation of human gastric cancer cell line SGC7901.

METHODSThe expressions of Vav3 proten in gastric cancer tissue, tumor-adjacent tissue, human gastric cancer cell line SGC7901 and gastric epithelial cell line GES-1 cells were tested by Western blot. Vav3-siRNA was transfected into the SGC7901 cells. The proliferation of SGC7901 cells in vitro was measured by MTT assay. Cell cycle of SGC7901 cells was determined by flow cytometry.The expressions of proliferation-related genes PCNA, p16, cyclin D1, Rb were determined by qPCR and Western blot assay. Orthotopic transplantation nude mouse models of gastric cancer were prepared, and the tumor growth and expressions of PCNA, P16, cyclin D1, and Rb proteins were examined.

RESULTSThe relative expressions of Vav3 in the gastric cancer and peritumoral tissue were 0.910±0.242 and 0.243±0.045, respectively; the relative expressions of Vav3 in SGC7901 and GSE-1 cells were 0.925±0.127 and 0.277±0.038, respevtively (both P<0.05). The expression of Vav3 protein in SGC7901 cells was effectively inhibited by Vav3-siRNA. Proliferation of SGC7901 cells was inhibited by (83.43±10.17)% after 80 nmol/L Vav3-siRNA transfection (P<0.05). The ratio of SGC7901 cells in G0/G1 phase was increased, and in S phase decreased after Vav3-siRNA transfection (both P<0.05). The expressions of PCNA and cyclin D1 were decreased in cells after Vav3-siRNA transfection, and expressions of p16 and Rb were increased after Vav3-siRNA transfection (P<0.05 for all). The tumor growth in the Vav3-siRNA group was much slower than that in the other 2 control groups of nude mouse models. Compared with the two control groups, expressions of PCNA and cyclin D1 were significantly lower in the Vav3-siRNA group, while expressions of p16 and Rb were increased (P<0.05 for all).

CONCLUSIONVav3 can promote the proliferation of gastric cancer cells by regulating proliferation-related genes.

Animals ; Cell Cycle ; Cell Line, Tumor ; Cell Proliferation ; Cyclin D1 ; metabolism ; Humans ; Mice ; Mice, Nude ; Proto-Oncogene Proteins c-vav ; metabolism ; RNA, Small Interfering ; Stomach Neoplasms ; metabolism ; Transfection