Objective To observe the expressions of microRNA-126 (miR-126) and vascular endothelial growth factor (VEGF) in gastric carcinoma tissues,and their associations with clinicopathologic features in gastric carcinoma.Methods Fifty cases of gastric carcinoma and matched tumor adjacent tissue specimens were collected from January 2012 to March 2014 in Shangqiu First People's Hospital of He' nan Province.Expression levels of miR-126 and VEGF were examined using reverse transcriptase-polymerase chain reaction.Their correlations and the relationships between them and the clinicopathologic features in gastric carcinoma were analyzed.Results The expression levels of miR-126 in the tumor tissues and in the adjacent tissues were 0.652 ± 0.102 and 0.379 ± 0.069,and the expression levels of VEGF in the two kinds of tissues were 1.523 ± 0.142 and 0.604 ± 0.152,with statistical differences (t =8.374,P =0.001;t =29.508,P =0.000).The expressions of miR-126 and VEGF were significantly positively correlated (r =0.735,P =0.001).The expressions of miR-126 and VEGF were associated with TNM stage (t =22.981,P =0.001;t =18.053,P =0.001),depth of tumor invasion (t =26.792,P =0.001;t =20.234,P =0.001) and tumor node metastasis (t=26.668,P=0.001;t=14.857,P=0.001).Conclusion miR-126 is high expressed in the gastric carcinoma tissue,which may up-regulate the expression of VEGF,and lead to the formation of new blood vessels,then to promote gastric carcinogenesis and the development process.

Objective To investigate the protective effect of mild hypothermia on cerebral ischemia-reperfusion injury in rats and the effect of mild hypothermia on the expression of inhibitor of differentiation 2 (Id2) protein.Methods A total of 72 adult male rats were randomly divided into a sham operation group,a normothermia group,and a mild hypothermia group.A model of middle cerebral artery occlusion was induced by a suture method.The mild hypothermia group was treated with low temperature (anal temperature 33±1 ℃,tympanic membrane temperature 31±1 ℃).Modified Neurological Severity Score (mNSS) was used to evaluate neurological deficits,triphenyltetrazolium chloride staining was used to detect infarct volume,and Western blot was used to detect the Id2 expression in the ischemic cortex at ischemia-reperfusion 6,12,24,and 72 h,respectively.ResultsThe mNSS scores in the mild hypothermia group were significantly lower than those in the normothermia group,the infarct volumes were significantly smaller than those in the normothermia group at ischemia-reperfusion 6,12,24,and 72 h (all P<0.001).Western blot analysis showed that the Id2 expressions in the ischemic cortex in the mild hypothermia group were significantly lower than those in the normothermia group at ischemia-reperfusion 6,12,24,and 72 h (all P<0.05).Conclusion s Mild hypothermia can decrease neurological deficits and reduce infarct volume after cerebral ischemia-reperfusion,its mechanism may be associated with the down-regulation of the Id2 expression.

Objective To investigate the expression of hypoxia inducible factor 1?(HIF-1?),vascular endothelial growth factor(VEGF) and micro vessel density(MVD) in gastric carcinoma and to explore their correlation with clinical pathological features such as cancer invasion and metastasis.Methods Forty-eight samples of gastric carcinoma tissues were examined for the expression of HIF-1?,VEGF and CD34 by immunohistochemical method.Results The positive expression rates of HIF-1? and VEGF were 66.7% and 60.4% in gastric carcinoma respectively.The mean value of MVD was 42.5?14.7 in gastric carcinoma.The expressions of HIF-1?,VEGF and the value of MVD were significantly correlated with the depth of invasion,lymph node metastasis and TNM stage.The HIF-1? expression was positively correlated with VEGF expression and MVD value.Conclusion The overexpression of HIF-1?,VEGF and MVD consist in gastric carcinoma tissue.The HIF-1? expression is positively correlated with VEGF expression and MVD value.The overexpression of HIF-1?,VEGF and MVD value are closely related with invasion,metastasis and poor biological behavior of gastric carcinoma.

Aim To investigate the mechanism for the inhibitory effect of hydrogen sulfide on the expression of tissue factor(TF)induced by oxidative low-density lipoprotein(ox-LDL)in endothelial cells.Methods Human umbilical vein endothelial cells (HUVECs ) were treated with 50 mg·L-1 ox-LDL in the absence or presence of different concentrations of NaHS (25 , 50,100 and 200 μmol·L-1 )for 24 h.The mRNA expression and protein content of TF in HUVECs were determined by reverse transcription PCR and ELISA, respectively.The content of intracellular reactive oxy-gen species (ROS)was determined by DCFH,an oxi-dative sensitive fluorescent indicator.The activation of nuclear factor-kappaB (NF-κB)was estimated by its expression in nuclear extracts analyzed by Western blot.Results Ox-LDL induced TF mRNA expression and increased TF protein content in HUVECs.The in-crease in intracellular ROS production and the activa-tion of NF-κB were observed in HUVECs treated with ox-LDL.However,NaHS could markedly inhibit the increases in TF mRNA and protein levels induced by ox-LDL.Also the elevation of intracellular ROS pro-duction and the activation of NF-κB elicited by ox-LDL were significantly suppressed by pretreatment with NaHS.In addition,pretreatment with BAY 1 1-7082 (10 μmol·L-1 ),the inhibitor of NF-κB or N-acetyl-L-cysteine(1 mmol·L-1 ),an antioxidant,could also decrease the TF mRNA and protein level as well as ROS production and NF-κB activation induced by ox-LDL in HUVECs,similar to the effects of 200 μmol· L-1 NaHS.Conclusion The mechanism for the in-hibitory effect of H2 S on the ox-LDL- induced TF ex-pression in endothelial cells may be related to inhibi-ting intracellular ROS production and subsequently NF-κB activation.

OBJECTIVETo obtain the peptide that specifically binds to human osteosarcoma MG-63 cells from Ph. D. 7TM phage display peptide library.

METHODSHuman osteosarcoma MG-63 cells were used as the target cells with human embryonic kidney 293T cells as the control for screening the peptide from Ph. D. 7TM phage display peptide library. The enriched specially binding peptides were verified by cell enzyme-linked immunosorbent assay (ELISA). The location of the peptide in MG-63 cells was investigated using cell fluorescence staining, and targeting of the peptide was tested by organ immunohistochemistry with Osteosarcoma model.

RESULTSThe specifically binding peptides were enriched after 4 rounds of screening. The sequence SLTNLSK was confirmed as the most frequent peptide by DNA sequencing and showed strong specificity verified by cell ELISA, fluorescent staining and organ immunohistochemistry.

CONCLUSIONA peptide that specifically binds to MG-63 cells has been screened from Ph. D. 7TM phage display peptide library to serve as a potential candidate for osteosarcoma-targeting therapy.

Amino Acid Sequence ; Animals ; Bone Neoplasms ; drug therapy ; Cell Line, Tumor ; Drug Screening Assays, Antitumor ; Humans ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Osteosarcoma ; drug therapy ; Peptide Library ; Peptides ; analysis ; Protein Binding

Objective To measure the ratio of the length between upper edge of eyebrow and edge of lower eyelid(L1)/the length between edge of lower eyelid and nasal base(L2)in the fordless-eyelid and upper lid ptosis patients who received levator aponeurosis folding surgery. To analyze the change in the proportion of midfa-cial aesthetics. Methods From December 2015 to October 2016,a total of 21 patients were recruited as study group. Those patientswere diagnosed with foldless eyelid and upper lid ptosis. They received levator aponeurosis folding procedures were carried out in our departmentand the pre- and post-operative photographs were collected. Full-face pictures of 20 Chinese female movie stars were prepared as control group. The ratiosof L1/L2(J)were measured via Adobe Photoshop CS6. The statistical significance was analyzed and the change of midfacial propor-tion was evaluated. Results In study group,the mean value of pre- and post-operative L1/L2(J1)is 0.746 and (J2)0.657 postoperatively. In control group,the mean value of L1/L2(J3)is 0.667. Statistical differences showed between the pre- and post-operative samples and between pre-operative samples and control group. There was no statistical difference between post-operative and control group samples. The ratios between L1 and L2 in post-opera-tive samples are close to those in control group ,which are closer to the golden radio. Conclusions Levator aponeurosis folding can effectively lower eyebrow in patients with upper lid ptosis and shorten the length between eyebrow and lower eyelid. Therefore harvesting a more appropriate and harmonious ratio of L1/L2 according the golden section theory. The changes in the proportion of midfacial aesthetics after operation could be used as a key point in the pre-operative communication and post-operative effect evaluation ,besides the size of palpebral fissure and corneal exposure rate.

Objective: To compare the treatment effect of surgical clipping and endovascular coil embolization for aneurysmal subarachnoid hemorrhage (aSAH) in the elderly. Methods: Elderly patients with aSAH (aged >65 years) treated in Shanghai Pudong Hospital from January 2009 to December 2017 were enrolled retrospectively. They were divided into craniotomy clipping group and endovascular intervention group according to the treatment strategy. The Glasgow Outcome Scale was used for short-term outcome assessment at discharge, 4-5 were defined as good outcome, and 1-3 were defined as poor outcome. Long-term follow-up was performed to assess clinical outcomes using the modified Rankin Scale, 0-2 was defined as good outcome and 3-6 were defined as poor outcome. The clinical and imaging information, perioperative complications, short-term and long-term clinical outcomes, and long-term imaging outcomes were compared between the two groups. Multivariate logistic regression analysis was used to assess the independent influencing factors of clinical outcomes. Results: A total of 68 elderly patients with aSAH were enrolled. Of these, 47 (69.1%) received endovascular coil embolization and 21 (30.9%) underwent microsurgical clipping. There were no significant differences between the two groups in the incidence of perioperative complications, short-term and long-term adverse outcomes, and imaging recurrence rate. Multivariate logistic analysis showed that smoking (odds ratio [OR] 36.319, 95% confidence interval [CI] 3.530-373.640; P=0.003), modified Fisher grade (OR 20.406, 95% CI 2.022-205.964; P=0.011) and World Federation of Neurological Societies (WFNS) grade (OR 4.686, 95% CI 1.012-21.692; P=0.048) were the independent risk factors for short-term poor outcomes in elderly patients with aSAH. Conclusion Both endovascular intervention and microsurgical clipping are safe and effective treatments for elderly patients with aSAH.

Objective:To explore the application of laparoscopic training in the standardized residency training of pediatric general surgery.Methods:A total of 36 rotating residents who received the standardized residency training in the department of pediatric general surgery of Shenzhen Children's Hospital from January 2017 to December 2019 were selected for laparoscopic training. The training content includes the study and training of laparoscopic theory knowledge, laparoscopic surgery video, skills operation of simulated operating system. Assessment was conducted before and after the training, and statistical analysis was performed to compare the difference of scores before and after the training. The self-evaluation of resident's learning efficiency and the satisfaction with teachers were investigated by questionnaires. SPSS 20.0 was used for paired t-test. Results:After receiving the standardized residency training of pediatric general surgery, the theoretical knowledge and cognition of the application on pediatric general laparoscopic surgery were significantly improved among the 36 residents. The time that the skills spent in vitro simulation box during the simulated operation training was significantly reduced after training, with a statistically significant difference ( P < 0.05). According to the questionnaire survey, resident's satisfaction with self-evaluation of learning efficiency was 97.22% (35/36), their satisfaction with teachers was 94.44% (34/36), and their satisfaction with teaching curriculum was 100.00% (36/36). Conclusion:Laparoscopic training can effectively improve the clinical practice ability of pediatric general surgery residents, which is worthy of promotion.

Objective To investigate the mechanism of iron death induced by TRPC6/NF-κB in glomerular podiocytes mediated by high homocysteine(Hcy).Methods Mouse glomerulopocytes were cultured in vitro and divided into Control group(0 μmol/L Hcy)and Hcy group(80 μmol/L Hcy).After 48h of intervention,Western blot was used to detect the expression levels of iron death related proteins GPX4 and SLC7A11 and TRPC6 and NF-κ B.Real-time quantitative fluorescence PCR(qRT-PCR)and immunofluorescence were used to detect the expression of TRPC6.The level of podocyte apoptosis was detected by flow cytometry.Malondialdehyde(MDA)assay kit was used to determine intracellular MDA levels.After transfection of TRPC6 interference fragment and TRPC6 negative control(NC),qRT-PCR was divided into Control,si-NC and si-TRPC6(Si-TRPC6-1,Si-TRPC6-2,Si-TRPC6-3).Western Blot was divided into Control,Hcy,si-NC+Hcy,si-TRPC6+Hcy.The expression of TRPC6 mRNA was detected by qRT-PCR.The expression levels of GPX4,SLC7A11,NF-κB and TRPC6 were detected by Western Blot.The level of podocyte apoptosis after interference was detected by flow cytometry.Results(1)Compared with Control group,the expression levels of iron death related proteins GPX4 and SLC7A11 in Hcy group were decreased,and the apoptosis rate was increased(P<0.05).(2)Compared with Control group,TRPC6 protein,mRNA levels and immunofluorescence expression were increased in Hcy group.The level of MDA and the expression of NF-κB signaling pathway protein increased in Hcy group,and the comparison between the two groups had statistical significance(P<0.05).(3)Compared with the si-NC group,the mRNA expression level of TRPC6 in si-TRPC6(Si-TRPC6-1,Si-TRPC6-2,Si-TRPC6-3)group was decreased,and the interference effect of Si-TRPC6-3 was the best(P<0.05).After transfecting TRPC6 NC and TRPC6 interference fragment and administering Hcy,there was no difference in GPX4,SLC7A11,NF-κB and TRPC6 expression in si-NC+Hcy group compared with Hcy group.Compared with the si-NC+Hcy group,the si-TRPC6+Hcy group had higher expression of iron death related proteins,GPX4 and SLC7A11,lower expression of NF-κB and TRPC6,and decreased apoptosis rate(P<0.05).Conclusion This study confirmed that TRPC6/NF-κB can regulate iron death of renal podocytes under the induc-tion of Hcy,which is one of the mechanisms leading to kidney injury.

Objective:To explore the in vitro anticancer effect of bleomycin (Ble) combined with cisplatin (DDP) on human gastric cancer cells.Methods:CCK-8 experiment was conducted to detect the effect of Ble and DDP alone or in combination on the proliferation of gastric cancer cells; Flow cytometry was used to detect changes in cell apoptosis and changes in reactive oxygen species. Cell scratch test was employed to detect cell migration; Western blot experiment was conducted to detect protein expression.Results:The results of CCK-8 showed that 20 μM DDP could inhibit the proliferation of six types of gastric cancer cells by 40%, while the effect of 20 μM Ble was only about 20%. When used in combination, the inhibitory rate on six types of gastric cancer could reach over 60%, with the strongest inhibitory effect on AGS cells. When 10 μM Ble and 10 μM DDP were combined, the proportion of AGS apoptotic cells reached 75.68%, higher than that of the two drugs treated with 20%, respectively 20 μM processing effect. When 10 μM Ble and 10 μM DDP were used in combination, the number of migrating and invading cells was significantly reduced, and the inhibition rate reached 50%. Compared with single use, the difference was statistically significant ( P<0.05). Flow cytometry results showed that the combination of Ble and DDP promoted cancer cell apoptosis by up-regulating the level of reactive oxygen species in cells; Western blot results showed that the expression of p-JAK2 and p-STAT3 decreased after Ble and DDP were combined, indicating that the JAK2/STAT3 signal pathway was inhibited. Conclusions:The combination of Ble and DDP can inhibit the proliferation and migration of gastric cancer cells, promote the apoptosis of gastric cancer cells, and play a synergistic anti-cancer effect. Its mechanism may be related to the up-regulation of ROS levels in cells, thereby inhibiting the activation of the JAK2/STAT3 pathway.