Objective To study the expression of inducible nitric oxide synthase (iNOS),endothelial nitric oxide synthase (eNOS) and vascular endothelial growth factor (VEGF) in human gastric cancer and their relationship with tumor angiogenesis and to investigate the interaction of NOS and VEGF in gastric cancer.Methods The expression and distribution of VEGF, iNOS and eNOS in 34 gastric cancer specimens were detected with immunohistochemistry. Microvessel density (MVD) was counted with FⅧRAg immune specific staining.Results The expression rates of iNOS, eNOS and VEGF in 34 gastric cancers were 73.5%, 82.4%and 91.2%respectively. The expression of VEGF had a significant positive relation with iNOS, but not with eNOS. The MVDs of VEGF or iNOS positive gastric cancers were obviously higher than those of VEGF or iNOS negative gastric cancers. There was no significant difference between the MVDs of eNOS positive gastric cancers and eNOS negative ones. Conclusion MVD increases with increase of expression of VEGF and iNOS in gastric cancer. It is indicated that VEGF and iNOS can promote gastric cancer angiogenesis. VEGF and iNOS have a significant positive correlation, which suggests that in human gastric cancer, iNOS plays an important role in the production and action of VEGF.

BACKGROUND: The implantation technique of bone marrow mesenchymal stem cells (MSCs) is of important significance for repair of brain injury. However, its action pathway still needs to be investigated.OBJECTIVE: To observe the changes of Bcl-2 and Bax expressions in the injured regions of cerebral cortex and hippocampus of rats with cerebral ischemia in which MSCs were implanted, and to analyze the action mechanism of intracranial implantation of MSCs inhibiting neuronal apoptosis.DESIGN: A randomized controlled experiment.SETTING: Institute for Cerebrovascular Disease, the Affiliated Hospital of Qingdao University Medical College.MATERIALS: Twenty-four male adult Wistar rats, weighing 180 to 240 g, were provided by the Experimental Animal Center of Shandong University. The involved rats were randomized into 3 groups with 8 in each: control group, injury group and implantation group. 5-bromodeoxyuridine (BrdU, Sigma Company), TUNEL kit, Bcl-2, Bax antibody kit were purchased from Beijing Zhongshan Bioengineering Company.METHODS: This study was carried out in the Institute for Cerebrovascular Disease, the Affiliated Hospital of Qingdao University Medical College between April 2005 and September 2006. Rats in the injury group and implantation group were developed into rat models of cerebral ischemia by suture of external carotid artery. Seven days later, the successful rat models in the implantation group were injected in the cerebral cortex and striatum with 2×1012 L-1 MSCs suspension primarily cultured in vitro. The processing of the experimental animals corresponded to the requests of Animal Ethics.MAIN OUTCOME MEASURES: Neuronal apoptosis and Bcl-2 and Bax expressions in the injured regions of cerebral cortex and hippocampus were detected by TUNEL staining and immunohistochemical method on the 7 and 14 days after successful modeling, separately.RESULTS: ①Neuronal apoptosis: On the 7th day after successful modeling, apoptotic cells were not found in the control group, and apoptotic cells in the implantation group were significantly fewer than those in the injury group (P ＜0.01). ② Bcl-2 and Bax expressions: On the 14th day after successful modeling, Bcl-2-positive neuronal expression in the cerebral cortex and hippocampus of rats in the injury group was significantly weaker than that in the control group and implantation group (P ＜ 0.01). Bax-positive expression in the cerebral cortex and hippocampus of rats in the injury group was significantly stronger than that in the control group and implantation group (P ＜ 0.01).CONCLUSION: MSCs can promote Bcl-2 expression and inhibit Bax expression of rats with cerebral ischemia injury,and accordingly neuronal apoptosis will be reduced.

Objective To study the apoptosis of endothelial cells and the relation between it and the expression of P53 protein after focal cerebral ischemia reperfusion in rats.Methods The apoptosis of endothelial cell 2,6,12 h and 1,2,3,7,14,21 d after cerebral ischemic reperfusion were observed using an in situ end labeling method,and the expression of P53 protein was detected by immunohistochemical staining methods.Results Apoptotic endothelial cells in ischemic penumbra were observed 2h after cerebral ischemic reperfusion, they peaked at 12~24 h, and decreased gradually.There was no remarkable difference between it and the sham operative group at 21 d.The P53 protein began to express 6h after cerebral ischemic reperfusion, it peaked at 1~2 d, and then declined gradually to controlled level at 7d. The expression peak time of P53 was 24 h later than that of cell apoptosis.Conclusion Apoptosis was a pattern of endothelial cell death after reperfusion of MCAO. P53 protein played an important role in the process of apoptosis of endothelial cells.

ObjectiveThis study was to evaluate the effects of different pneumoperitoneum used in laparoscopy on in vitro tumor cells growth. Methods 3H-TdR incorporation was used to compare the DNA synthesis (CPM value) of tumor cells after exposure to simulated laparoscopic environments, composed of carbon dioxide, helium and room air, respectively. Results At 24 hours after pneumoperitoneum, compared with group of helium, air and control, CPM increased in group of carbon dioxide (F=47.576 5,P0.05). At 48,72 and 96 hours, pneumoperitoneum, compared with helium group, air group and control group, CPM increased significantly in carbon dioxide group (F=116.183,1082.92 and 5116.75, P0.05). Conclusion Carbon dioxide promotes tumor cells growth, while helium restrains the growth of tumor cells. [

Objective:To analyze the clinical characteristics of primary acute myeloid leukemia (AML) (non-M3 type) in children suffering from different levels of platelet count(PLT).Methods:In the Tumor Hospital of Zhengzhou University from January 2014 to December 2018, laboratory and clinical data of 247 de novo primary AML pediatric patients were retrospectively reviewed.According to the PLT before treatment, patients were divided into very low platelet group (VLG), low platelet group (LG) and non-lowing platelet group (NLG), with<50×10 9/L, ≥50×10 9/L but <125×10 9/L and ≥125×10 9/L as the boundaries.All patients were followed up until June 30, 2019.Meanwhile, the follow-up data was obtained by consulting medical records or by telephone.SPSS 17.0 software was applied for data analysis. Results:In general clinical features, a different group of hemoglobin (Hb) content, fusion gene AML- ETO and clinical risk stratification were statistically significant in different PLT groups ( χ2=11.270, 12.115 and 12.848, respectively, all P<0.05). However, the differences of other indicators in different groups of PLT were not statistically significant (all P>0.05). There were no statistically significant differences in terms of 3-year disease-free survival(DFS) rate (59.3%, 36.3%, 50.4%) among the 3 groups (all P>0.05). The median total survival(OS)time(40.5 months)and 3-year OS rate(41.0%) of NLG patients were significantly higher than those of VLG(23.1 months, 30.1%)and LG(14.1 months, 18.2%)patients, with statistically significant differences( χ2=7.798 and 6.553, respectively, all P<0.05). The univariate analysis of gender, white blood cell(WBC), Hb, PLT, lactic dehydrogenase(LDH), FLT3-ITD, NPM1, DNMT3A, CEPBA, C-KIT, AML-ETO, molecular genetic prognosis, complete remission(CR), and hemopoietic stem cell transplantation(HSCT) displayed that DNMT3A mutation was an adverse factor that affects patients′ OS ( χ2 =5.834, P<0.05), and the positive factors that influences OS were non-reducing PLT before treatment, and obtaining CR and subsequent HSCT ( χ2=7.798, 79.168, and 31.337, respectively, all P<0.05). Multi-factor analysis revealed that the independent protective factors that affect patients′ OS were the non-reducing PLT before treatment, and obtaining CR and subsequent HSCT( Wald=42.760, 15.918, and 10.183, respectively, all P<0.05). Conclusions:Before treatment, non-reducing PLT is a protective factor for primary childhood AML patients, and the prognosis is satisfying.

Object To determine the content of glycyrrihizic acid obtained when each individual ingredient in MAHUANG DECOCTION was decocted separately and then mixing the extracts with boiling water in comparison with that obtained by decocting the total composition together as a whole in the traditional way. Methods The contents of glycyrrhizic acid was determined by HPLC. Hypersil C 18 column was used, with acetonitrile∶0.1% acetic acid (33∶67) as the mobile phase and detected at the wavelength of 254 nm. Results The average recovery of glycyrrhizic acid when separately decocted was 102.43%, RSD=2.65%, while that of decoction in whole was 99.41%, RSD=3.11%. Conclusion The method was simple and accurate and was not interfered by other constituents in the prescription.

Objective: To compare the contents of ferulic acid in single medicine decoction (preparing the refined granules of single traditional Chinese herb from the different kinds of medicinal materials separately then mixing the granules with boiling water) and mixed medicine decoction (decocting the all kinds of the medicinal materials together as the traditional method) of Tangkuei Blood Supplementing Decoction (composed of Radix Angelicae Sinensis and Radix Astragali). Methods: HPLC conditions: Hypersil C 18 column methanol -0.05% acetic acid solution (45∶55) as mobile phases, detection wavelength at 324 nm. Results: The average recovery of single medicine decoction was 100.86% with RSD =2.33%( n =6), and that of mixed medicine decoction was 101.37% with RSD =1.52%( n =6).Conclusion: The content of ferulic acid mixed medicine decoction was higher than that in single medicine decoction.

Objective To investigate the feasibility of a rat carotid atherosclerosis model induced by artery clamping with high-cholesterol diet and vitamin D3.Methods Twenty Wista rats were randomly divided into either a general diet group (n =5) or a high cholesterol diet group (n =15).After clamping the right common carotid arteries of the rats in the high cholesterol diet group,they were fed with high fat diet,and vitamin D3 (600 000 IU/kg) was injected intraperitoneally.The lipid levels of the general diet group and the high cholesterol diet group were detected at 6 and 12 weeks respectively.The bilateral cormmon carotid arteries were selected for preparing paraffin sections and were stained with HE staining.The pathological changes in blood vessels were observed.Results The levels of serum total cholesterol at 12 weeks (2.803 ± 1.307 mmol/L vs.1.513 ±0.281 mmol/L; P=0.017) and low-density lipoprotein cholesterol (0.660± 0.260 mmol/L vs.0.311 ±0.078 mmol/L; P =0.003) in the high-cholesterol diet group were significantly higher than those at 6 weeks.The histopathological examination showed that the common carotid artery intimas on the clamping sides were incomplete,the foam cell deposition was observed under intima,the atherosclerotic plaques or fibrous plaques were observed on the surface of cavity,inside the plaques contained necrotic tissue,and thrombosis was observed in the cavity.The common carotid artery intima-media thickness in the general diet group (n =5) at 12 weeks was 8.3 ± 1.1 μm.The sham-operated sides (n =20) and clamping sides (n =20) were 8.8 ± 0.7 μm and 97.4 ±25.7 μm,respectively.There were significant differences among the three groups (F =116.313,P=0.000).The clamping sides in the high-cholesterol diet group were significantly higher than the shamoperated sides in the high-cholesterol diet group (P=0.000) and the general diet group (P =0.000).Conclusions Common carotid artery clamping with high-cholesterol diet and vitamin D3 is a simple and feasible method for inducing a rat carotid atherosclerosis model.

Objective To measure the organ weights , blood physiological and biochemical parameters of KK /Upj-Ay/J.Methods KK/Upj-Ay/J mice at five and ten weeks of age were selected , and the organ weights , blood physiological and biochemical parameters were observed .Results Parts of the organ weights , blood physiological and biochemical parameters of different ages and sexes were significant differences .The fasted blood glucose of KK/Upj-Ay/J mice reached 7.0mmol/L at 10 weeks of age .Conclusion The results show that the organ weights , blood physiological and biochemical parameters are affected by age and gender of KK /Upj-Ay/J mice.The fasted blood glucose reached the diabetes level at 10 weeks of age .

AIM To establish an ultra-performance liquid-chromatography tandem mass spectrometry (UPLC-MS/MS) method for the simultaneous content determination of five constituents in Jiangzhi Huoxue Tablets (Polygoni multiflori Radix,Astragali Radix,Chuanxiong Rhizoma,etc.).METHODS The 50％ methanol extract of this drug was performed on a 40 ℃ thermostatic Restek UItra BiPh column (100 mm × 2.1 mm,5 μm),with the mobile phase comprising of acetonitrile (containing 0.1％ formic acid)-0.1％ formic acid flowing at 0.4 mL/min in a gradient elution manner.RESULTS Stilbene glycoside,tanshinone Ⅱ A,emodin,ferulic acid and puerarin showed good linear relationships within the ranges of 4.01-1 027,0.7-187,1.48-380,3.98-1 020 and4.285-1 097 ng/mL (r ＞0.994 0),whose average recoveries were 98.57％-101.0％ with the RSDs (n =6) of less than 4.79％.CONCLUSION This specific and sensitive method can be used for the quality control of Jiangzhi Huoxue Tablets.