ObjectiveStudies have shown that nuciferine has anti-cerebral ischemia effect， but the specific mechanism of action has not been elaborated. Based on the transcriptome results， the pharmacological mechanism of nuciferine against cerebral ischemia was analyzed from multiple dimensions including tissue， cell， pathological process， biological process and signaling pathway. MethodsThirty SD rats were randomly divided into the sham group， model group and nuciferine group（40 mg·kg^-1） according to weight. Except for the sham group， the model of middle cerebral artery occlusion（MCAO） was established by thread embolization method after 30 min of administration in the other two groups. Twenty-four hours after surgery， transcriptome sequencing was used to detect the gene expression profiles in the cortex penumbra of rat cerebral tissue， and gene ontology（GO） and kyoto encyclopedia of genes and genomes（KEGG） pathway enrichment analysis were performed for differentially expressed genes. The mechanismof nuciferine against cerebral ischemia was analyzed from 5 dimensions of tissue， cell， pathological process， biological process and signaling pathway by the transcriptome-based multi-scale network pharmacology platform（TMNP）. ResultsTranscriptome sequencing and gene quantitative analysis showed that 667 genes were significantly reversed by nuciferine. Further enrichment analysis of KEGG and GO suggested that the pathways of nuciferine involved regulating stress response， ion transport， cell proliferation and differentiation， and synaptic function. TMNP research found that at the tissue level， nuciferine could significantly improve the cerebral tissue injury caused by ischemia. At the cellular and pathological levels， nuciferine could play an anti-cerebral ischemia role by improving the state of various nerve cells， mobilizing immune cells， regulating inflammation. And at the level of biological processes and signaling pathways， nuciferine mainly acted on the processes such as vascular remodeling， inflammation-related signaling pathways， and synaptic signaling. ConclusionCombined with the results of transcriptome sequencing， gene quantitative analysis and TMNP， the mechanism of nuciferine against cerebral ischemia may be related to processes such as intervening in stress response and inflammation， affecting vascular remodeling and regulating synaptic function. These results can provide a basis and reference for further study of the pharmacological mechanism of nuciferine against cerebral ischemia.

Objective:To study the relation ship between the branch patterns of inferior mesenteric artery (IMA) and imaging pelvic measurement parameters for anastomotic leakage (AL) after anterior resection (AR) of rectal cancer.Methods:Five hundred thirty-four patient were enrolled from Jan 2008 to Dec 2018 at the General Surgery Department of Guizhou Provincial People's Hospital. The AL related imaging risk factors were analyzed by chi-square test or Fisher's exact test.Results:AL was found in 36 (6.7%) patients. AL related mortality rate was 11.1% (4/36) compared to 0.4% (2/498) in those without the complications of no AL cases ( P<0.001). Seven pelvic imaging measurement results were attained in 412 patients including anteroposterior diameter of the inlet of the pelvis, anteroposterior diameter of the outlet of the pelvis, upper edge of the symphysis pubis to the tip of the coccyx, sacrococcygeal distance angle from the lower edge of the pubis to the upper edge of the pubis to the sacral promontory, distance between the ischial spines and that of ischial tuberosity. Univariate analysis showed that there was no significant relationship between the above 7 pelvic measurement parameters and the occurrence of AL (all P>0.05). There was no significant relationship between branch patterns of IMA and AL after rectal cancer surgery ( P=0.712). Conclusion:AL as a severe postoperative complication in rectal cancer patients undergoing AR procedure were caused by multiple factors. Neither IMA branch patters nor pelvic imaging measurement seem to be related to the occurrence of AL after AR for rectal cancer.

Objective:To analyze the clinicopathological features and prognostic factors of alpha‐fetoprotein‐producing gastric carcinoma (AFPGC).Methods:A retrospective analysis was made on 2 671 GC patients admitted from Jan 1998 to Dec 2018 , AFPGC patients and matching AFP negative GC cases were enrolled and their clinicopathological features and prognostic factors were analyzed. The survival curve was drawn by Kaplan-Meier method. Log-rank test was used to test the significance, Univariate analysis was performed by using COX proportional hazard model.Results:There were 98 AFPGC in this study accounting for 4.5% of all GC of the corresponding time period. The proportion of male to female was 2.16∶1, the average age was (65±12) years. The serum AFP levels significantly decreased after operation in most patients (median: 52 ng/ml vs. 5 ng/ml, Ｚ=-2.736, P=0.001). Serum AFP and CEA levels in patients with AFPGC before treatment were significantly higher than that in patients with AFP negative GC (both P<0.05) . Vascular invasion(62.71% vs. 40.68%) and liver metastasis (31.63% vs .6.12%) were more likely to occur in AFPGC groups (both P<0.05). However, there was no significant difference between the two groups in tumor size, location, differentiation and lymph node metastasis (all P>0.05). The prognosis of AFPGC was significant pooer than that in AFP negative GC ( P<0.05). Prognosis of AFPGC patients was significantly correlated with preoperative serum AFP level, TNM stage, lymph node metastasis, simultaneous liver metastasis and vascular invasion (all P<0.05) . COX multivariate survival analysis found that preoperative serum AFP level was independent risk factors of patients with AFPGC ( P<0.05). Conclusion:AFPGC is a special GC charactering poor prognosis .

Objective To investigate the expression of macrophage migration inhibition factor (MIF) and cell cycle regulating factor Cyclin D1 in hepatocellular carcinoma tissue and the interaction between MIF and Cyclin D1 in hepatocellular carcinoma cell cycle controlling. Methods Using quantitative real-time PCR and Western blotting to detect mRNA and protein expression of MIF and Cyelin DI in HCC tissues and tumor adjacent tissues. Specific small interfering RNA(siRNA) targeting MIF gene was transfccted at doses of 50 nmol/L and 100 nmoL/L into HCC cell lines of PLC and HepG2 with lipofeetamine 2000 methods to knockdown the expression of M1F gene and to investigare the the interaction between M1F and Cyclin D1. Results MIF and Cyclin D1 protein and mRNA were overexpressed in HCC tumor tissues. The relative expression of MIF,Cyclin D1 protein and mRNA were 0.825±0.13,0.843± 0.104 and 7.31±1.85 folds、4.27±1.05 folds, compared with the tumor adjacent tissues (FMIF= 15.5, P<0.01;FCyclin D1=87.5,P <0.01). In MIF siRNA treated PLC and HepG2 cells, MIF mRNA down regulation 71.2%±7.2%, 87.4%±2.9% ,74.3%±8.9% and 88.4%±4.6% respectively (FPLC = 315.5 ,P < 0.01 ; FHepG2= 201.2 P < 0.01). While MIF protein expression were significandy reduced to 0.33±0.03,0.11±0.02, 0.81±0.08 and 0.36±0.02 in a dose-dependent manner (FPLC= 43.9, P <0.01 ;FHepG2 = 133.4 P <0.01). Cyclin D1 mRNA was significantly down-regnlated in MIF siRNA treated PLC and HepG2 cell lines when compared with control group(P <0.01). In 50 nmol/L and 100 nmol/L groups, Cyclin DI mRNA levels were respectively decreased by 68.2%±3% and 78.1%±1.4% in PLC cell, 65.8%±4.7% and 77.3%±2.6% in HepG2 cell (FPLC= 1569, P < 0.01 ; FHepG2= 480.4, P <0.01). Compared with control groups, Cyclin D1 protein levels significantly reduced to 0.28±0.06、0.15±0.03 and 0.44 ±0.04、0.13±0.02 in the PLC and HepG2 after M IF siRNA treatment(FPLC= 35.5, P < 0.01 ; FHepG2 = 114.7, P < 0.01). Conclusions MIF and Cyclin D1 mRNA and protein were overexpressed in HCC tumor tissues and participated in tumor cell cycle regulation. MIF may up-regnlate the expression of Cyclin DI via ERK signalling and precipitate in carcinogenesis of hepatocellular carcinoma.

Objective To observe inhibiting effect of endostatin on subcutaneous transplant tumor of breast carcinoma, and to illuminate the therapeutic effect of endostatin in the cancer. Methods The effect of en-dostatin on MCF-7 cell proliferation was studied by MTr. The model of MCF-7 cell transplant tumor on nude mice was constructed. Endostatin was injected intradermally around the transplant tumor. Inhibition effect on the tumor was observed. Results Endostatin with the concentration of 10 μ/mL and 15 μg/mL can inhibitMCF-7 cell proliferation effectively (P < 0. 05 ). After endostatin injection, tumor weight, volume and mi-crovessel density decreased significantly(P < 0.05 ). Conclusion Endostatin can inhibit breast carcinoma proliferation through inhibiting angiogenesis and the tumor cell itself.

Objective To investigate vascular endothelial growth factor-c (VEGF-C) and vascular endothelial growth factor receptor-3(VEGFR-3) mRNA expression, microvessels density (MVD) and lymphatic microvessels density (LVD) in human hepatocellular carcinoma and normal liver tissue. Try to illuminate the relationship among VEGF-C,VEGFR-3,MVD,LVD and the clinical pathological features of hepatocellular carcinoma. Methods Liver tissue of 60 cases definitely diagnosed as hepatocellular carcinoma and 20 normal cases were collected. VEGF-C and VEGFR-3 mRNA expression were examined by RT-PCR, MVD and LVD were examined by immunohistochemistry staining. Relationship between these indexes and clinical pathological features of hepatocellular carcinoma was also analysed. Results VEGF-C and VEGFR-3 mRNA expression, MVD and LVD in hepatocellular carcinoma were higher than those in normal liver tissue (P<0.01); In hepatocellular carcinoma tissue, expression of VEGF-C mRNA positively related with VEGFR-3 mRNA, MVD and LVD(P<0.01). VEGF-C and VEGFR-3 expression positively related with portal vein tumor thrombus, intrahepatal metastasis and lymph node metastasis (P<0.01). MVD positively related with portal vein tumor thrombus and intrahepatal metastasis (P<0.01). LVD positively related with lymph node metastasis (P<0.01). Conclusion VEGF-C and VEGFR-3 expression increase in hepatocellular carcinoma tissue. They might play roles in tumor invasion and metastasis by inducing angiogenesis and lymphangiogenesis.

Objective To investigate the effects of short hairpin RNA (shRNA) expression plasmid targeting vascular endothelial growth factor-C (VEGF-C) on the proliferation and invasion of HepG2 cells. Methods The VEGF-C shRNA plasmid vector labeled with green fluorescent protein was constructed and stably transfected into HepG2 cells. The transfected cells were sorted by G418 and visualized by fluorescent microscope and assayed by flow cytometry. Expression of VEGF-C in transfected cells was determined by RT-PCR and Western blot. The inhibition rates of the cell proliferation and invasion were determined by MTT assay and reconstituted basement membrane invasion assay, respectively. Results VEGF-C shRNA effectively downregulated VEGF-C mRNA and protein expression in HepG2 cells, and it also effectively inhibited the proliferation of HepG2 cells in a time-dependent manner. The invasion capacity of HepG2 cells was inhibited by VEGF-C shRNA, and the inhibition rate was 51.54%. Conclusions VEGF-C plays an important role in tumor proliferation, invasion and metastasis. RNA interfering technology that targets VEGF-C may serve as a potential therapeutic intervention in the treatment of human hepatic cancer.

Objective To investigate the efficacy of bend-assisted laparoseopie fight bemicolectomy for right colon carcinoma in elderly obesity patients. Methods 20 obesity eases undergoing hand-assisted laparoscopic right bemicolectomy were retrospectively reviewed, and compared with 25 obesity cases with transabdominal right hemicolectomy in the same period. The safety, recovery, eradication and stress reaction were compared. Results In hand-assisted laparoscopic group, there were less bleeding loss, rapid recovery to normal temperature and gastrointestinal function,ont of bed activity, short-time hospitalization compared with transabdominal group(P <0.01). There were no differences in operative time, numbers of lymph nodes removed, postoperative complications and length of specimen between two gronps(P > 0.05). CRP, IL-6, adrenocorticotrophic hormone (ACTH) and cortisol were more significantly increased in two groups after operation than before operation (P < 0. 05) , and this increase was moreprominent in transabdominal group than in hand-assisted laparoacopic group (P < 0.05). Conclusion Hand-assis-ted laparoscopic right hemicolectomy is a safe and effective way for elderly obesity patients,especially in postoperative recovery and physical stress reaction.

Objective To evaluate the impact of the recombined plasmid vector with enhanced green fluorescent protein (EGFP) encoding soluble tumor necrosis factor related apoptesis inducing ligand (pIRES-EGFP-sTRAIL) on proliferation and apoptosis of human hepatocellular carcinoma cell line HepG2, and investi-gate the feasibility and efficiency of the transfection of pIRES- EGFP- sTRAIL into HepG2 by ultrasound micro-bubble contrast agent. Methods pIRES-EGFP-sTRAIL was constructed and transfected into HepG2 cells by using different types of mediated methods: microbubble echocontrast agent combining appropriate dose of ultra-sound irradiation, liposome method, microbubble echocontrast agent only or blank medium treatment. Transfec-tion efficiency was evaluated by EGFP-expressed cell count; proliferation-lnhibiting rate and the apoptosis rate of HepG2 cells were determined by MTT method and flow cytometry analysis; changes of cell morphology were examined by microscopy with Hoechst33258 dyeing; expression of caspase-8 and caspase-3 was detected by Western blot. Results Ultrasound microbubbh enhanced pIKES-EGFP-sTRAIL uptake by HepG2 cells, and the transfection efficiency was significantly higher in ultrasound microlmbble group than that in other groups( P<0.05 ) ; pIRES- EGFP- sTBAIL effectively inhibited HepG2 cell proliferation and induced cell apoptosis by triggering caspase cascade. Both the inhibiting rate and apoptosis rate were significantly higher in ultrasound microbubble group than those in other groups(P<0.05). Conclusion pIRES-EGFP-sTRAIL expresses ef-fectively in HepG2 cells, sTRAIL has a potential role on the inhibiting proliferation and inducing apoptosis of HepG2 cells by triggering caspase cascade, and this role can be enhanced by the administration of low-intensity ultrasound and microbubble echecontrast agent.

Objective To investigate the anti-tumor effect of the recombined adeno-associated virus encoding melanoma differentiation -associated gene-7 (MDA-7) regulated by progression-elevated gene (PEG) promotor on human hepatocellular carcinoma (HCC) in nude mice. Methods A nude mouse model of subcutaneously implanted HCC cell line HepG2 tumor was established. AAV-PEG-MDA-7 was injected from the tail vain after tumor cell innoculation. RT-PCR, Western blot and immunohistochemical analysis were employed to detect MDA-7 expression in mice; MDA-7 plasma concentration was detected by ELISA assay. Tumor growth was observed, tumor cell apoptosis and angiogenesis in tumor tissues were measured by TUNEL and immunohistochemical analysis. Results Seven days after tumor cell innoculation RT-PCR, Western blot, and immunohistochemistry showed that MDA-7 was only expressed in the liver. ELISA assay showed that the concentration of MDA-7 in plasma was gradually increased to reach the plateau (200 ng/ml). Tumor growth was significantly inhibited in mice injected with rAAV-PEG-MDA-7, and the tumor growth-inhibiting rate was 62%. TUNEL and immunohistochemical analysis demonstrated significant induction of tumor cell specific apoptosis and reduction of vascular formation in tumor tissues. Conclusions rAAV-PEG-MDA-7 exhibits tumor-specific cytotoxicity and liver tendency, inhibiting tumor growth possibly by tumor cell apoptosis-induciug effect and antiangiogenesis.