In order to understand the difference and expression of genes in CEK cells before and af-ter baicalin interferes with IBV,further reveal and analyze the mechanism of baicalin interfering IBV replication in CEK cells in vitro.After IBV infected CEK cells for 2 h,9.75 mg/L baicalin liq-uid was added to interfere with CEK cells,which was recorded as the baicalin(H-IBV)group,and three replicate wells were set in the control group and the IBV(IBV)infection group.After 36 h culture,cell samples were collected and subject to transcriptome for sequencing.The results showed that there were 102 differentially expressed genes in H-IBV group compared with IBV in-fection group,among which 48 genes weresignificantly up-regulated and 54 genes were significantly down-regulated.Through functional annotation in GO and KEGG databases,it was found that dif-ferentially expressed genes were mainly annotated in biological processes such as cellular proces-ses,biological regulation,metabolism,and secondary pathways such as viral infectious diseases,signal transduction and interaction.Retinol metabolism pathway,phospholipid transfer to mem-brane,IL-27 mediated signaling pathway,MDA5/RIG-I and Toll-like receptor signaling pathway were significantly enriched in CEK cells,and the production of type Ⅰ interferon and interferon al-pha and the process of antiviral infection were also positively regulated.There were more differen-tial genes enriched in nucleic acid catalysis,immune system,and reaction,and interbiological reac-tion.Through the STRING network interaction map,it was found that most immune-related genes could form a 36-node interaction network centered on IRF7,TLR3 and STAT1.Therefore,com-pared with IBV group,the differentially expressed genes after baicalin treatment were mainly an-notated and enriched in the biological process,and the immune system and response were en-hanced,mainly through the positive regulation of IRF7 in the MDA5/RIG-I receptor signaling pathway and the inhibition of TLR3 signal transduction in the Toll-like receptor signaling path-way.Positive regulation of IL-27 mediated pathway and regulation of JAK-STAT signaling path-way were supplemented by activation of the expression of IRF7,TLR3,STAT1 and other related genes,and interaction with corresponding downstream proteins to promote the expression of IFN-α and regulatory cytokines,coupled with negative regulation of viral(defense)response and viral processes.Thus,baicalin interferes with IBV replication in CEK cells.

Objective:To evaluate the efficacy of a modified continuous penetrating-suture pancreaticojejunostomy (PPJ), also known as a continuous PPJ with a U-shaped reinforcement of pancreatic section (U-PPJ), in patients with high risk of clinically relevant postoperative pancreatic fistula (CR-POPF).Methods:Clinical data of 33 patients with pancreatic tumors undergoing pancreatic surgery in the Department of Hepatobiliary Surgery, the First Affiliated Hospital of Wannan Medical College from August 2017 to December 2023 were collected, including 22 males and 11 females, aged (64.9±8.6) years old. According to the fashion of pancreaticojejunostomy, patients were divided into two groups: U-PPJ group ( n=11) and PPJ group ( n=22). The general data, incidence of CR-POPF, abdominal bleeding and other clinicopathological data were collected. Results:All patients underwent pancreatic surgery successfully and were discharged from the hospital uneventfully. Intraoperative blood loss in U-PPJ group was 200.00 (100.0, 200.0) ml, postoperative hospitalization was 13.0 (11.0, 18.0) d, and the drain removal time was 17.0 (12.0, 21.0) d, and no CR-POPF occurred. The intraoperative blood loss, postoperative hospitalization days, drain removal time, and incidence of postoperative biochemical leakage were comparable between the groups (all P>0.05). The incidence of CR-POPF in U-PPJ group was lower than that in PPJ group [0 vs. 22.7% (5/22), P<0.05]. Conclusion:U-PPJ is safe and effective in patients with pancreatic tumors and might reduce the incidence of CR-POPF.

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.

Objective:To observe the changes of insulin secretion in the early stage of severe scald in rats, and to explore its signal transduction mechanism.Methods:Twenty-four male Wistar rats aged 7 weeks were divided into sham injury alone (SIA) group, sham injury+ BPV (HOpic) (SIB) group, scald alone (SA) group, and scald+ BPV (HOpic) (SB) group using the random number table, with 6 rats in each group. Full-thickness scald of 50% total body surface area was inflicted in rats of SA and SB groups by a 6-s immersion of the abdomen and a 12-s immersion of the back in 94 ℃ hot water. Rats in SIA and SIB groups received sham injuries through immersion of the back and abdomen in 37 ℃ warm water for 6 and 12 seconds respectively. From 0 (immediately) to 2 day (s) after injury, the rats in groups SB and SIB were intraperitoneally injected with the phosphatidylinositol 3-kinase/protein kinase B (PI3K/Akt) signaling pathway enhancer BPV (HOpic) solution (0.5 mg/mL) at the dosage of 0.6 mg/kg once a day, and the rats in groups SA and SIA were intraperitoneally injected with the same volume of dimethyl sulfoxide once a day. At post injury hour (PIH) 72, the tail blood of rats was sampled for measuring fasting blood glucose (FBG) with a glucometer, and the pancreatic tissue samples of rats was harvested for observing the pathological manifestations of islets by hematoxylin-eosin staining, counting the docked granules per 10 μm membrane of islet beta cells and calculating the proportion of insulin vesicles through the observation of the ultrastructure of islet beta cells by transmission electron microscope, and detecting the phosphorylation level of Akt in the pancreatic PI3K/Akt signaling pathway by Western blotting. Data were statistically analyzed with one-way analysis of variance and least significant difference test.Results:(1) At PIH 72, the rat FBG levels in SIA and SIB groups were normal and similar ( P>0.05). Compared with the levels of those two groups, the rat FBG level in SA group was increased significantly ( P<0.01), while the level in SB group showed no obvious change ( P>0.05). Compared with that in SA group, the rat FBG level in SB group was decreased significantly ( P<0.01). (2) At PIH 72, the morphology of rat islets was complete and the islet cells distributed regularly in SIA and SIB groups. Compared with those in SIA and SIB groups, the morphology of rat islets was incomplete, the insulin vesicles in islets were common, the islet cells distributed irregularly, and the cytoplasm of some islet beta cells was lightly stained or translucent in SA group; the morphology of islets in SB group did not change obviously. Compared with those in SA group, the morphology of islets was comparatively complete, the insulin vesicles in islets were less common, the islet cells distributed comparatively regularly, and the lightly stained or translucent cytoplasm of islet beta cells was less in SB group. (3) At PIH 72, the number of docked granules per 10 μm membrane of rat islet beta cells and the proportion of insulin vesicles in SIA and SIB groups were similar ( P>0.05). Compared with those in SIA and SIB groups, the number of docked granules per 10 μm membrane of rat islet beta cells in SA group was decreased significantly ( P<0.01), while the proportion of insulin vesicles was increased significantly ( P<0.01); the number of docked granules per 10 μm membrane of rat islet beta cells in SB group was obviously decreased ( P<0.05), while the proportion of insulin vesicles did not change obviously ( P>0.05). Compared with those in SA group, the number of docked granules per 10 μm membrane of rat islet beta cells in SB group was significantly increased ( P<0.01), while the proportion of insulin vesicles was significantly decreased ( P<0.01). (4) At PIH 72, the phosphorylation levels of Akt in SIA, SIB, SA, and SB groups were 0.91±0.03, 0.98±0.03, 0.78±0.08, and 0.87±0.08, respectively. Compared with that in SIA group, the phosphorylation level of Akt was increased obviously in SIB group ( P<0.05) but was decreased significantly in SA group ( P<0.01), while the level in SB group did not change obviously ( P>0.05). Compared with the level in SIB group, the phosphorylation levels of Akt in SA and SB groups were decreased significantly ( P<0.01). Compared with that in SA group, the phosphorylation level of Akt in SB group was increased significantly ( P<0.05). Conclusions:At the early stage post severe scald in rats, the activity of the pancreatic PI3K/Akt signaling pathway and the function of insulin secretion are reduced. Improving the activity of the pancreatic PI3K/Akt signaling pathway in rats can ameliorate the function of insulin secretion and recover the physiological level of blood glucose.

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 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.

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

Objective The purpose of this study was to investigate the effect and mechanism of Vav3 gene on the proliferation of human gastric cancer cell line SGC7901.Methods The 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.Results The 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) .Conclusion Vav3 can promote the proliferation of gastric cancer cells by regulating proliferation-related genes.