Objective To explore the impact of using large dose norepinephrine on the anastomotic healing after esophagectomy.Methods Clinical data of data of 148 cases patients with esophageal cancer who were treated in Wuwei Tumor Hospital of Gansu Province with surgery from January 2014 to June 2016 were retrospectively analized.There were 7 patients who were used 25-67 mg ((32.6 ± 2.3) mg) norepinephrine because of the low blood pressure and other factors during the first 4 d postoperative as research group,and the other 141 patients who had not applied or applied the small dose were set as control group.The patients of two groups with postoperative anastomotic healing and fistula incidence were compared.Results There were 3 cases and 7 cases of postoperative patients appeared anastomotic fistula in the research group and the control group,the rate of anastomotic fistula were 42.86％ (3/7) and 4.96％ (7/141) respectively,the difference was significant (x2=9.78,P =0.001),and 2 cases appeared varying degrees residual gastric necrosis in the research group,all of them occurred in the patients with large dosage and long time.Conclusion There are great risk on the anastomotic fistula and residual stomach mecrosis if long time and large dose norepinephrine was used after esophagectomy,it should be caused enough attention for surgeons.

Objective To observe the effects of different concentrations of baicalin on the mouse model of experimental autoimmune encephalomyelitis (EAE) and to explore its mechanisms.Methods A mouse model of EAE was established with MOG33-55 peptide and bacillus Calmette-Guerin (BGG) vaccine with complete Freund adjuvant (CFA).At the third day after immunization, high and low doses of baicalin were administered to the mice intragastrically once a day for 20 days.The neurological function of mice was evaluated.TUNEL staining was used to detect apoptosis in the spinal cord tissue.The level of ATP in spinal cord tissue was detected by an ATP determination kit.Furthermore, the protein expressions of Bax, Bcl-2, cleaved cas-3 and cleaved cas-9 were detected by western blot, respectively.Results Baicalin improved the neurological function and delayed the onset time in EAE mice.

To elucidate the molecular characteristics of the hemagglutinin (HA) genes of H5N1 subtype of avian influenza viruses in the boundary region of Yunnan province. Of 420 samples were collected from the foreign poultry in boundary region of Yunnan province during 2003 to 2008 and these samples were subjected to screening by H5/N1 subtype-specific and multiplex RT-PCR. testing. The HA genes of H5N1 viruses from positive samples were amplified by RT-PCR and cloned into vector pMD18 T for subsequent sequencing. The alignment with sequences of the known reference strains and phylogenetic analysis were then performed. The genes from 21 representative positive samples with 4 different sequences at the cleavage site were obtained and all of them possessed the molecular characteristic of highly pathogenic avian influenza virus. The mutation of key amino acids had been found among receptor-binding sites, potential glycosylation sites and neutralizing epitopes.-Phylogenetic analysis showed those positive samples could be divided into 5 distinct clades, including clade 1, 2.4. 2.3.2, 2.3.4 and 7. It is evident that H5N1 viruses from the foreign boundary region of Yunnan province in 2003 to 2008 show genetic divergence and clade 2,3,4 is the dominant clade in this region.

ed in a Chinese pedigree with HHD.

Objective:To investigate the molecular mechanism of antiglioma effect of curcumin.Methods:Cell experiment: (1) U251MG and SHG-44 cells at logarithmic growth phase were treated with 10 μmol/L curcumin (curcumin group) or same volume of dimethyl sulfoxide solution (control group); cells were transfected with negative control small interfering RNA (siRNA) and long non-coding RNA (lncRNA) H19 siRNA (negative control siRNA group and H19 siRNA group); cells were transfected with negative control siRNA and H19 siRNA, respectively, and then, they were treated with 10 μmol/L curcumin (negative control siRNA+curcumin group and H19 siRNA+curcumin group); the H19 siRNA was co-transfected with negative control miR inhibitor or miR-491-5p inhibitor into these cells (H19 siRNA+negative control inhibitor group and H19 siRNA+miR-4915p inhibitor group); H19 siRNA+negative control miR inhibitor or H19 siRNA+miR-491-5p inhibitor were co-transfected into the cells, and then, they were treated with 10 μmol/L curcumin (H19 siRNA+negative control inhibitor+curcumin group and H19 siRNA+miR-491-5p inhibitor+curcumin group); the cells were co-transfected with miR-491-5p mimic+blank plasmid or miR-491-5p mimic+HOXA9 overexpression plasmid, and then they were treated with 10 μmol/L curcumin (miR-491-5p mimic+blank plasmid+curcumin group and miR-491-5p mimic+HOXA9 overexpression plasmid+curcumin group); real-time fluorescent quantitative PCR (qRT-PCR) was used to detect the mRNA expressions of H19, miR-491-5p, and HOXA9; CCK-8 assay was used to detect the cell proliferation; flow cytometry was used to detect the cell apoptosis; plate cloning method was employed to detect the number of cell clone formation; Transwell assay was used to detect the cell migration; and the HOXA9 protein expression was measured by Western blotting. (2) The 293T cells at the logarithmic growth phase were chosen; the negative control miRNA mimics or miR-491-5p mimics combined with wild-type H19, wild-type HOXA9 3'-UTR plasmid vectors were co-transfected into the cells, respectively (negative control mimic+wild type H19 group and miR-491-5p mimic+wild type H19 group, negative control mimic+wild type HOXA9 3'-UTR group and miR-491-5p mimic+wild type HOXA9 3'-UTR group); the luciferase activity was detected by dual luciferase reporter experiment. (3) Thirty specimens from glioma patients (glioma group) underwent surgical resection and pathologically confirmed in our hospital from May 2017 to May 2019 and 30 normal brain tissue specimens obtained during decompression (normal group) at the same period were chosen; the mRNA expressions of H19, miR-491-5p, and HOXA9 were detected by qRT-PCR, and the HOXA9 protein expression level in these specimens was detected by Western blotting. (4) Twenty-four nude mice were randomly divided into negative control short hairpin RNA (shRNA) group, H19 shRNA group, negative control shRNA+curcumin group, and H19 shRNA+curcumin group ( n=6); U251MG cells stably transfected with negative control shRNA or H19 shRNA were intraperitoneally injected, respectively, into the mice; and 60 mg/kg curcumin was injected on the next d; the tumor volume was measured on the 7 th, 11 th, 15 th, 19 th, 23 rd, and 27 th d of rearing; and the H19, miR-491-5p and HOXA9 mRNA expressions in the tumor tissues were detected by qRT-PCR; the HOXA9 protein expression was detected by Western blotting. Results:(1) When curcumin group comparing with control group, and H19 siRNA group comparing with negative control siRNA group, U251MG and SHG-44 cells had significantly decreased miR-491-5p mRNA and protein expressions, and significantly increased miR-491-5p mRNA expression ( P<0.05); as compared with that in the H19 siRNA+negative control inhibitor group, the HOXA9 mRNA and protein expressions in U251MG and SHG-44 cells of H19 siRNA+miR-491-5p inhibitor group were significantly higher ( P<0.05). When curcumin group comparing with control group, H19 siRNA group comparing with negative control siRNA group, H19 siRNA+curcumin group comparing with negative control siRNA+curcumin group, the U251MG and SHG-44 cells after 72 h of culture had significantly decreased cell proliferation rate, significantly increased apoptosis rate, significantly reduced number of cell clone formation, and significantly reduced cell migration number ( P<0.05). When H19 siRNA+miR-491-5p inhibitor+curcumin group comparing with H19 siRNA+negative control inhibitor+curcumin group, miR-491-5p mimic+HOXA9 overexpression plasmid+curcumin group comparing with miR-491-5p mimic+blank plasmid+curcumin group, the U251MG and SHG-44 cells after 72 h of culture had significantly increased cell proliferation rate, significantly reduced apoptosis rate, significantly increased number of cell clone formation, and significantly increased cell migration number ( P<0.05). (2) When miR-491-5p mimic+wild-type H19 group comparing with negative control mimic+wild-type H19 group, miR-491-5p mimic+wild-type HOXA9 3'-UTR group comparing with negative control mimic+wild-type HOXA9 3'-UTR group, the cell luciferase activity was significantly reduced ( P<0.05). (3) As compared with those in the normal group, the H19 and HOXA9 mRNA expressions and HOXA9 protein expression in the glioma group were significantly increased, and the miR-491-5p mRNA expression was significantly reduced ( P<0.05). (4) On the 27 th d of rearing, when H19 shRNA group comparing with negative control shRNA group, and H19 shRNA+curcumin group comparing with negative control shRNA+curcumin group, the tumor volume was significantly reduced, the miR-491-5p mRNA expression in the tumor tissues was significantly increased, and the H19 mRNA, HOXA9 mRNA and protein expressions were significantly reduced ( P<0.05). Conclusion:Curcumin may inhibit the cell proliferation and migration and promote the apoptosis of glioma cells through lncRNA H19/miR-491-5p/HOXA9 axis.

Objective:To investigate the effects of high-energy shock and vibration on cortex injury and peripheral blood immune cells in goats.Methods:Seventeen Boer goats without gender preference were selected. By using random number tables, the goats were divided into normal control group ( n=5) and shock and vibration injury group ( n=12). The goats in the normal control group were anatomized routinely and their brain was collected after being sacrificed without any other treatment. The goats in the high-energy shock and vibration model group were placed on a loading table (part of the BY10-100 instant shock and vibration simulation platform) in a restrained state, and made into a high-energy shock and vibration injury model induced by a vertical impact waveform generator. The intravenous blood samples were taken from the goats in the shock and vibration injury group before and at 0, 3, 6 and 24 hours after injury.Then, the goats were sacrificed and the following procedures were the same as the normal control group. At 24 hours after injury, the brain injury and the histopathological changes of the cerebral cortex in the normal control group and shock and vibration injury group were observed by gross pathological and anatomical examination and HE staining. The mRNA expression of zonula occludens 1 (ZO-1), tight junction protein 5 (Claudin-5), glial fibrillary acidic protein (GFAP), ionized calcium binding adaptor molecule 1 (IBA-1), interleukin (IL)-1β, IL-6 and cluster of differentiation antigen 177 (CD177) of the cerebral cortex in the normal control group and shock and vibration injury group were measured through fluorogenic quantitative polymerase chain reaction. The expression of ZO-1 and Claudin-5 proteins of the cerebral cortex in the normal control group and shock and vibration injury group were detected by Western blotting. Hematology analyzer and coagulation analyzer were used to detect white blood cell count, neutrocyte, lymphocyte, monocyte, prothrombin time 1 (PT-1), prothrombin time international normalized ratio (PT-INR), activated partial thromboplastin time (APTT), thrombin time (TT), prothrombin activity (PTA) and fibrinogen (FIB) levels in goats of the shock and vibration injury group before and at 0, 3, 6 and 24 hours after injury, respectively. Results:At 24 hours after injury, no visible contusion or necrosis was found in goat brain tissue in the shock and vibration injury group; the cerebral micro-vessels presented with a local dilation, hyperemia, edema, aggregation of inflammatory cells, disruption of vessel walls and leakage of red blood cells. These changes were not observed in the normal control group. In the shock and vibration injury group, ZO-1 and Claudin-5 mRNA expressions in the cerebral cortex were 0.25±0.10 and 0.09(0.04, 0.44) respectively, which were significantly lower than those of the normal control group [1.00±0.15 and 0.99(0.80, 1.20)]; GFAP, IBA-1, IL-1β, IL-6 and CD177 mRNA expression levels were 4.40(3.88, 6.75), 2.60±1.07, 3.04±0.51, 2.71±0.45 and 2.93±0.62 respectively, which were significantly higher than those of the normal control group [1.00(0.78, 1.22), 1.00±0.37, 1.00±0.27, 1.00±0.57 and 1.00±0.35]; ZO-1 and Claudin-5 protein expression levels were 0.41±0.06 and 0.42±0.11 respectively, which were significantly lower than those of the normal control group (1.08±0.12 and 0.91±0.23) (all P<0.01). In the shock and vibration injury group, the levels of white blood count, neutrocyte, and lymphocyte in peripheral blood were (13.7±3.3)×10 9/L, (35.3±14.8)% and (57.2±15.1)% respectively before injury, (19.4±3.1)×10 9/L, (60.5±12.5)% and (33.6±14.2)% respectively at 3 hours after injury, and (20.6±3.6)×10 9/L, (63.6±13.0)% and (30.9±15.0)% respectively at 6 hours after injury. By contrast, the levels of white blood count and neutrocyte were significantly increased but the level of lymphocyte was significantly decreased at 3 and 6 hours after injury ( P<0.05 or 0.01); the levels of the above indicators showed no significant changes at 0 and 24 hours after injury (all P>0.05); the level of monocyte did not change significantly at all time points before and after injury (all P>0.05). The levels of PT-1, PT-INR, APTT, TT, PTA and FIB in the shock and vibration injury group did not change significantly at each time point before and after injury (all P>0.05). Conclusion:Cerebral cortex microvascular injury and disruption of blood-brain barrier can be initiated in the early stage of high-energy shock and vibration injury in goats, accompanied by the presence of central and peripheral inflammatory response.

AIM: To investigate the relationship between vascular smooth muscle cell (VSMC) injury, organelle stress response and autophagic cell death (autophagy) and ferroptosis induced by the chemical hypoxia inducer cobalt chloride (CoCl2) through the bioinformatics analysis and in vitro cell experimentation. METHODS: The dataset GSE119226 of VSMC treated with cobalt chloride was acquired from the gene expression database (GEO). The R language was used to investigate the relationship between CoCl2 treatment and organelle stress response (Golgi stress, endoplasmic reticulum stress) and two forms of cell death (ferroptosis and autophagic cell death). With primary cultured rat VSMC (rVSMC) and CoCl2-induced anoxia model, the changes in cell viability were detected by CCK-8 method, and reactive oxygen species (ROS) levels were measured using DCFH-DA method. The expression levels of HIF-1α (a key molecule in hypoxia), Golgi stress markers GM130 and p115, endoplasmic reticulum stress markers GRP78 and CHOP, autophagy markers LC3-II / LC3-I and Beclin1, and ferroptosis markers GPx4 and xCT were detected by Western blot. The effect of inducing or inhibiting organelle stress and cell death on the CoCl2-induced cell damage was also observed. RESULTS: Differentially expressed genes analysis of GSE119226 dataset showed that CoCl2 treatment of VSMCs had significant effects on organelle function and stress response, autophagy and ferroptosis-related genes, in which endoplasmic reticulum stress, protein processing in endoplasmic reticulum, regulation of Golgi to plasma membrane protein transport, autophagy / autophagic cell death, and ferroptosis pathways were remarkably enriched. The results of in vitro experiment showed that compared with normal rVSMC, cell viability was significantly decreased after CoCl2 treatment, as well as HIF-1α protein expression and ROS levels in rVSMCs were increased. In rVSMC treated with Co-Cl2, the expression levels of Golgi structural proteins GM130 and p115 (reflecting the occurrence of Golgi stress) were decreased, while the markers GRP78 and CHOP (reflecting the occurrence of endoplasmic reticulum stress) were increased. At the same time, CoCl2 treatment also reduced the expression of autophagy markers LC3-II/LC3-I and Beclin1 (indicating the decrease levels of autophagy), while the expression of ferroptosis markers GPx4 and xCT were decreased (indicating the occurrence of ferroptosis). Compared with CoCl2 treatment group, induced Golgi stress, endoplasmic reticulum stress, or ferroptosis could further reduce cell viability, while inhibition of these processes could improve cell viability. On the other hand, increasing the level of autophagy can improve the cell viability. CONCLUSION: Hypoxia induced by cobalt chloride can lead to VSMC injury. Golgi stress, endoplasmic reticulum stress, ferroptosis, and the reduction of autophagy level play an important role in it. Inhibition of organelle stress response and ferroptosis, or increase of autophagy level can improve VSMC injury caused by cobalt chloride.