Objective:To evaluate the long-term outcomes and prognostic factors of locally advanced gastric cancer with serosa-invasion.Methods:This study is a retrospective cohort study. The clinical and pathological data of 495 patients with locally advanced gastric cancer with serosa-invasion who underwent laparoscopic radical gastrectomy in Department of General Surgery, the First Hospital Affiliated to Army Medical University from October 2012 to October 2018 was analyzed retrospectively. There were 356 males and 139 females with an age ( M(IQR)) of 59 (16) years (range: 18 to 75 years). Observation indicators included postoperative results and long-term prognosis. The survival curve was drawn by the Kaplan-Meier method. Univariate and multivariate prognostic analysis was performed using the Cox proportional hazards model. Results:Among the 495 patients, a total of 57 patients (11.5%) were lost to follow-up, with a follow-up time of 89 (40) months (range: 23 to 134 months). The 5-year disease-free survival rate (DFS) and the 5-year overall survival rate (OS) were 56.0% and 58.2%, respectively. The 5-year DFS for patients with stage ⅡB, ⅢA, ⅢB, ⅢC were 71.2%, 60.5%, 51.6%, 33.3%, respectively. The 5-year OS for patients with stage ⅡB, ⅢA, ⅢB, ⅢC were 71.2%, 62.2%, 54.1%, 39.3%, respectively. Multivariate analysis showed that age >65 years (DFS: HR=1.402, 95% CI: 1.022 to 1.922, P=0.036; OS: HR=1.461, 95% CI: 1.057 to 2.019, P=0.022), lymph node dissection number less than 25 (DFS: HR=1.348, 95% CI: 1.019 to 1.779, P=0.036; OS: HR=1.376, 95% CI: 1.035 to 1.825, P=0.028), pathological stage Ⅲ (DFS: HR=2.131, 95% CI: 1.444 to 3.144, P<0.01; OS: HR=2.079, 95% CI: 1.406 to 3.074, P<0.01), and no postoperative chemotherapy (DFS: HR=3.127, 95% CI: 2.377 to 4.113, P<0.01; OS: HR=3.768, 95% CI: 2.828 to 5.020, P<0.01) were independent prognostic factors for the decrease in DFS and OS rates. Conclusions:Laparoscopic radical gastrectomy for locally advanced gastric cancer with serosa-invasion could achieve satisfactory long-term oncological outcomes. More lymph node dissection and standardized postoperative adjuvant chemotherapy are expected to further improve the prognosis of patients with locally advanced gastric cancer with serous invasion after laparoscopic radical surgery.

Objective:To evaluate the long-term outcomes and prognostic factors of locally advanced gastric cancer with serosa-invasion.Methods:This study is a retrospective cohort study. The clinical and pathological data of 495 patients with locally advanced gastric cancer with serosa-invasion who underwent laparoscopic radical gastrectomy in Department of General Surgery, the First Hospital Affiliated to Army Medical University from October 2012 to October 2018 was analyzed retrospectively. There were 356 males and 139 females with an age ( M(IQR)) of 59 (16) years (range: 18 to 75 years). Observation indicators included postoperative results and long-term prognosis. The survival curve was drawn by the Kaplan-Meier method. Univariate and multivariate prognostic analysis was performed using the Cox proportional hazards model. Results:Among the 495 patients, a total of 57 patients (11.5%) were lost to follow-up, with a follow-up time of 89 (40) months (range: 23 to 134 months). The 5-year disease-free survival rate (DFS) and the 5-year overall survival rate (OS) were 56.0% and 58.2%, respectively. The 5-year DFS for patients with stage ⅡB, ⅢA, ⅢB, ⅢC were 71.2%, 60.5%, 51.6%, 33.3%, respectively. The 5-year OS for patients with stage ⅡB, ⅢA, ⅢB, ⅢC were 71.2%, 62.2%, 54.1%, 39.3%, respectively. Multivariate analysis showed that age >65 years (DFS: HR=1.402, 95% CI: 1.022 to 1.922, P=0.036; OS: HR=1.461, 95% CI: 1.057 to 2.019, P=0.022), lymph node dissection number less than 25 (DFS: HR=1.348, 95% CI: 1.019 to 1.779, P=0.036; OS: HR=1.376, 95% CI: 1.035 to 1.825, P=0.028), pathological stage Ⅲ (DFS: HR=2.131, 95% CI: 1.444 to 3.144, P<0.01; OS: HR=2.079, 95% CI: 1.406 to 3.074, P<0.01), and no postoperative chemotherapy (DFS: HR=3.127, 95% CI: 2.377 to 4.113, P<0.01; OS: HR=3.768, 95% CI: 2.828 to 5.020, P<0.01) were independent prognostic factors for the decrease in DFS and OS rates. Conclusions:Laparoscopic radical gastrectomy for locally advanced gastric cancer with serosa-invasion could achieve satisfactory long-term oncological outcomes. More lymph node dissection and standardized postoperative adjuvant chemotherapy are expected to further improve the prognosis of patients with locally advanced gastric cancer with serous invasion after laparoscopic radical surgery.

Objective:To investigate the effect and of piperine(PIP)on nerve injury in rats with cerebral infarction.Meth-ods:The rat model of cerebral infarction was prepared by modified thread embolism method.The rats were divided into Sham group,Model group,piperine group(PIP group,20 mg/kg piperine),piperine+PKA inhibitor group(PIP+H-89 group,20 mg/kg PIP+5 mg/kg H-89).The rats were scored for neurological impairment,the volume of cerebral infarction,neuronal damage and neuronal apoptosis were observed,and the levels of cAMP,IL-1β and IL-6 in hippocampus and the protein expressions of GFAP,NSE,AC6,PKA,p-CREB,CREB and BDNF were detected.Results:Compared with Sham group,Model group had large cerebral infarction,cerebral in-farction volume,the neurological deficit score,neuronal apoptosis rate,the levels of IL-6 and IL-1β in hippocampus,and number of GFAP positive cells,the positive expression rates of NSE protein increased,the number of Nissl bodies,and the expressions of cAMP,AC6,PKA,p-CREB and BDNF proteins in hippocampus decreased(P<0.05);compared with Model group,cerebral infarc-tion volume,the neurological deficit score,neuronal apoptosis rate,the levels of IL-6 and IL-1β in hippocampus,and number of GFAP positive cells,the positive expression rates of NSE protein in the PIP group decreased,the number of Nissl bodies,and the ex-pressions of cAMP,AC6,PKA,p-CREB and BDNF proteins in hippocampus increased(P<0.05);the effect of piperine in improving nerve injury in rats with cerebral infarction was able to be reversed by H-89.Conclusion:PIP can inhibit the apoptosis of hippocampal neurons,and improve the neural injury induced by cerebral infarction by activating AC/PKA/CREB signal pathway.

Objective:To investigate whether lncRNA LINC00909 affected the radiosensitivity of colorectal cancer cells by targeting miR-548-3p.Methods:The expression levels of LINC00909 and miR-584-3p in colorectal cancer and adjacent tissues were detected by qRT-PCR. The colorectal cancer cells SW480 and SW620 were cultured in vitro, and transfected with si-NC, si-LINC00909, miR-NC, miR-584-3p mimics, si-LINC00909, and anti-miR-NC and si-LINC00909, and anti-miR-584-3p, respectively. The cells were irradiated with a dose of 4 Gy. The cell survival fraction and sensitization enhancement ratio (SER) were detected by clone formation assay. Cell proliferation was detected by MTT assay. Cell migration and invasion were assessed by Trans well chamber assay. The targeting relationship between LINC00909 and miR-584-3p was confirmed by dual luciferase reporter assay. The effect of interfering with the expression of LINC00909 or inhibiting the expression of miR-584-3p on the weight of the xenograft tumor after irradiation was evaluated by subcutaneous xenograft experiment in nude mice. Results:The expression level of LINC00909 in colorectal cancer tissues was significantly up-regulated ( P<0.05), whereas the expression level of miR-584-3p was significantly down-regulated ( P<0.05). After interfering with the expression of LINC00909 or miR-584-3p overexpression, the cell survival fraction score was significantly reduced ( P<0.05), the SERs were 2.017 and 1.762, and cell proliferation, migration and invasion were suppressed (all P<0.05). Dual luciferase reporter assay confirmed that LINC00909 could target and bind to miR-584-3p. After interfering with the expression of LINC00909, the weight of the transplanted tumor was significantly reduced ( P<0.05), whereas the weight of the transplanted tumor was significantly increased after co-transfection with anti-miR-584-3p ( P<0.05). Conclusion:Interfering with the expression of LINC00909 can inhibit the proliferation, migration and invasion ability of colorectal cancer cells by up-regulating the expression of miR-548-3p, thereby enhancing the cell radiosensitivity.

Objective: To investigate the effect and potential mechanism of HOXC8 on the radiosensitivity of non-small cell lung cancer cell line A549, aiming to provide novel ideas for clinical combined treatment. Methods: The A549 cells with stable knockdown of HOXC8 were constructed by using lentivirus and validated by qPCR and Western blot. The radiosensitivity of A549 stable cell line was assessed by plate clone formation assay. The expression levels of TGF-β₁ and the proteins in the downstream signal pathway after knockdown of HOXC8 were detected by Western blot. Results: The A549 cells with stable knockdown of HOXC8 were successfully constructed. The viability and clonogenic capacity of A549 cells were significantly reduced after silencing HOXC8. Silencing HOXC8 also increased the sensitivity of A549 cells to radiotherapy and significantly inhibited the expression of TGF-β₁ and p-Smad2/3 proteins in the downstream signaling pathway. Conclusion Silencing HOXC8 can increase the sensitivity of A549 cells to radiotherapy probably by inhibiting TGF-β₁ signaling transduction. HOXC8 might play an important role in A549 cells.

To establish a method for isolation, culture and identification of adipose-derived mesenchymal stem cells (ASCs) from the inbreed line miniature pig of Wuzhishan (ILMW).
 Methods: A total of 100 g adipose tissues were obtained from subcutaneous tissues of neck in six-month old healthy ILMW (3 samples, male). ASCs from ILMW (ILMW-ASCs) were isolated from adipose tissues through 0.1% collagenase digestion. The cells at the 3rd, 5th, 8th, 13th passages were collected. Cell morphology, size, phenotype, cell cycle, and apoptosis were monitored. Cell differentiation was induced and cell proliferation curve was drawn.
 Results: The ILMW-ASCs, fibroblast-like or whirlpool-like, began the adherence at 36 h and entered a logarithmic phase in the 5th day. Eighty percent of them were fused in the 7th day. The average diameter and volume of ILMW-ASCs were (17.00±0.54) µm and (2.58±0.24)×10-9 L, respectively. The expressions of CD29, CD44 and CD90 were positive, and there was no significant difference between the different passages (all P>0.05). The expressions of CD45, CD8a and HLA-DR were increased with the increase in passages after the 3th passage (all P<0.05). The adipogenic induction of ILMW-ASCs was observed by positive oil red O staining, and the osteogenic induction of ILMW-ASCs was determined by positive alizarin red staining. Apoptosis and senescence occurred in the 13 passage of ILMW-ASCs, and the proportion of S phase of cell cycle was lower than that in lower passages (all P<0.05). 
 Conclusion: ILMW-ASCs are one of the best choice for porcine ASCs, which might provide a source of candidate stem cells for therapy of large animal disease models and tissue or organ repairment.
Adipose Tissue ; Animals ; Cell Differentiation ; Cells, Cultured ; Male ; Mesenchymal Stem Cells ; Swine ; Swine, Miniature

Objective To investigate the effect of lncRNA CCAT1 and miR-130b-3p on the radiosensitivity of human pancreatic cancer cells PANC-1.Methods Real-time PCR was used to detect the relative expression levels of CCAT1 and miR-130b-3p in pancreatic cancer tissues and cell lines including PANC-1 cells irradiated with 2 Gy X-rays.After silencing CCAT1 and/or inhibiting miR-130b-3p expression,cell apoptosis rate,Caspase 3 activity and cell survival were detected by flow cytometry,Caspase 3 activity detection kit and colony formation assay,respectively.Cell survival curve was stimulated by the multi-target single-hit model.Based on the starBase v2.0 online analysis,the luciferase reporter gene assay,RNA-binding protein immunoprecipitation assay (RIP) and Real-time PCR assay were applied to verify the relationship between CCAT1 and miR-130b-3p.Results CCAT1 expression was up-regulated (t=6.322-8.555,P＜0.05),but miR-130b-3p expression was down-regulated (t =3.950-18.795,P＜ 0.05) in the radiation-resistant pancreatic cancer tissues,pancreatic cancer cell lines and 2 Gy-irradiated PANC-1 cells.When the CCAT1 silenced PANC-1 cells were irradiated with 2 Gy,cell survival fraction decreased (t=2.929,5.047,5.234,5.125,P＜0.05),apoptosis rate and Caspase 3 activity increased (t=6.953,6.836,P＜0.05).CCAT1 could selectively regulate miR-130b-3p expression.Inhibition of miR-130b-3p expression could enhance PANC-l cell survival (t =4.564,6.736,8.656,P＜0.05),but reduced apoptosis rate (t=5.234,P＜0.05) and Caspase 3 activity (t=10.440,P＜0.05).Conclusions Silencing CCAT1 promotes the expression of miR-130b-3p and enhances radiosensitivity of PANC-1 cells.

Objective To investigate the change of CD35 expression on neutrophils in the peripheral blood and the relationship between the change and disease activity in patient with myeloperoxidase antineutrophil cytoplasmic antibody-associated vasculitis (MPO-AAV).Methods Forty untreated patients with active MPO-AAV(patient group)and forty healthy volunteers (control group) were enrolled into this study,and Bermingham vasculitis activity score (BVAS) for every patient was recorded.Flow cytometry (FCM) was employed to detect the CD35 and MPO expression on the neurtrophil,and enzyme linked immunosorbent assay (ELISA) was taken to test the levels of autoantibody against MPO-Antineutrophil cytoplasmic antibody (MPO-ANCA),fragment a from the activated complement factor B (Ba) and MPO in peripheral blood from both group.All test results were compared between the 2 groups by t test,Non-parametric test,Spearman correlation analysis.In addition,the relations among the laboratory results and the relationship between BVAS and the laboratory results were analyzed respectively.Results Compared with the control group,the expression level,which was represented as mean flourscence indensity (MFI),of CD35 and neutrophil membrane MPO on peripheral blood neutrophils was significantly increased [(2 014±968) vs (1 454±511),t=3.024,P=0.002 and (709±244) vs (580±158),t=2.806,P＜0.01,respectively],and the MPO expression level in neutrophils was significantly lower [(1 525±1 033) vs (3 196±2 126),t=-4.468,P＜0.01].Ba and MPO levels in serum of the patient group was significantly higher than that in the control group [37.89(26.17,63.14) μg/L vs 27.99(18.64,46.52) μg/L,Z=-2.521,P=0.012 and 546.16(450.55,729.96) U/L vs 327.93(279.02,365.10) U/L,Z=7.121,P＜0.01,respectively].In patient group,the expression level of CD35 had a significant positive relationship with peripheral blood neutrophil count (r=0.573,P＜0.01),serum Ba (r=0.433,P=0.005) and BVAS (r=0.368,P=0.020),respectively,whereas,there was a negative correlation between the MPO expressed on the neutrophils and that in the neutrophils (r=-0.458,P=0.003),and a positive relationship between MPO-ANCA and BVAS (r=0.351,P=0.026).Conclusion There is significant increased expression of CD35 on the neutrophil of patient with MPO-AAV,which might protect the neutrophil from destruction by the activated complement alternative pathway,and more neutrophils consequently contribute to the MPO-AAV pathogenesis.Inhibition of CD35 expression might become one of the potential new pathways for the treatment of MPO-AAV.

Objective To analyze the exposed dose of hippocampus(HC)of T3,T4nasopharyngeal carcinoma patients treated with intensity modulated radiotherapy(IMRT). Methods The bilateral HCs were delineated and were divided into head(HH),body(HB)and tail(HT)for 62 nasopharyngeal carcinoma patients treated with IMRT.The dose parameters of HC were then analyzed. Results The mean dose of left and right HC was(1 127±704)cGy,(1 173±762)cGy. The mean dose of left HH,HB and HT was(1 732±1029)cGy,(820±632)cGy,(423±366)cGy(P=0.000);while the mean dose of right HH, HB and HT was(1 985±1101)cGy,(837±531)cGy,(432±343)cGy(P=0.000).The exposed dose and the volume exposed in different dose of HH were obviously higher than those of HB and HT.The dose parameters of HH,HB and HT decreased in turn. The involvement of sphenoid sinus,ethmoid sinus and cavernous sinus correlated with high exposed dose of HC. Conclusions The exposed dose of HH,HB and HT was different in nasopharyngeal carcinoma patients treated with IMRT.The exposed dose of HH was the highest,which should be emphasized especially. The involvement of sphenoid sinus,ethmoid sinus and cavernous sinus suggest high exposed dose of HC.

Objective To explore preliminarily the role of the E2 subunit of pymvate dehydrogenase (PDC-E2) modified by xenobiotics (e.g.2-octynic acid,2-OA) in the pathogenesis of primary biliary cirrhosis (PBC).Methods Patients of PBC (102 cases),primary sclerosing cholangitis (PSC,34 cases) and healthy controls (HC,50 cases) were selected.The anti-PDC-E2,anti-2-OA and anti-lipoic acid (LA) antibody in the peripheral blood of the 3 groups were tested by enzyme linked immunosorbent assay (ELISA).By inhibitive ELISA (iELISA),30 of the 102 PBC patients with anti-PDC-E2 antibody but without anti-2-OA antibody were selected to detect whether there was any new epitope on the PEC-E2 conjugated with 2-OA.The chi-square test and Fisher exact test were taken to analyze the enumeration data.The two-tailed unpaired t test with Welch's correction was used to compare the measurement data.Spearman rank correlation analysis was also employed for proper test.Results The positive rate of anti-PDC-E2,anti-LA and anti-2-OA antibody in PBC patients was 94.1％(96/102),73.5％(73/102) and 53.9％(55/102) respectively,all of which were statistically significantly higher than those in healthy controls group but were of no significant difference between PSC and healthy controls group.There was no significant relevance between the levels of Anti-LA and anti-2-OA antibody in the PBC group (r=-0.065,P=0.520).The iELISA results showed that the antibody,which only identified the epitopes on 2-OA-PDC-E2 induced by the 2-OA conjugation with PDC-E2,existed in 40％(12/30) of the PBC patients,and more interestingly,this antibody was predominantly appeared in PBC patients at their early clinical stage.Conclusion There are anti-LA antibody and anti-2-OA antibody in PBC patients,which have shown no significant association with each other.It is very likely that new antigenic conformational epitopes on PDC-E2 modified by 2-OA would emerge,which might led to the immune response in the individuals who are susceptible to PBC,and thus contribute for the breaking of PDC-E2 immune tolerance,and PBC occurrence finally.