The myocardial protective effect of continuous infusion with normothermic oxygenated crystalloid solution or blood cardioplegia during normothermic CPB was studied in 15 dogs. Ultrastructure. levels of adenine nucleotides,lipid peroxide (LPO),water content of heart musle,hemodynamics were observed. Dogs were randomly divided into three groups. Group Ⅰ: intermittent infusion with cold crystsalloid cardioplegia during hypothermic CPB (n=5); Group Ⅱ: continuous infusion with normothermic oxygenated blood cardioplegia during normothermic CPB(n=5); Group Ⅲ: continuous infusion with normothermic oxygenatde crystalloid cardioplegia duriug normothermic CPB((n=5). The normal mitochandria contents and glycogen stores in group Ⅱ and Ⅲ were significantly higher than those in group Ⅰ(P

Objective To discuss the spiral CT features of lymph node metastasis in advanced gastric cancer, and to investigate the correlation between spiral CT features and pathologic results. Methods Spiral CT scan and triphasic enhancement spiral CT scan were performed in 59 patients with advanced gastric cancer before operation. Results ①The pathologic metastatic rate of lymph node in gastric cancer was high when the lymph nodes were nubby-mixed, prominently enhanced, unsymmetrical enhanced and ≥9 mm in diameter on spiral CT (P<0.05). ②The detection rate of lymph node metastasis with spiral CT was high when carcinoma tissues were poorly differentiated, Borrmann Ⅲ+Ⅳ, T3-4 staging and TNM Ⅲ+Ⅳ staging. Conclusion The spiral CT features of lymph node (distributing type, size, enhanced degree, enhanced mode and staging) can reflect the pathologic characteristics of lymph node metastasis in gastric carcinoma. Pathologic characteristic of gastric tumor determines the detection rate of lymph node metastasis on spiral CT. The accuracy of CT to diagnose lymph nodes metastasis will be improved by integrating the spiral CT features of both gastric tumor and lymph node.

Objective To investigate the inhibitory effects of docosahexaenoic acid (DHA)and 5-fluorouracil (5-FU)in combination on human gastric cancer cell line AGS in vitro .Methods Human gastric cancer line AGS was treated with different concentrations of DHA and 5-FU alone or in combination.The inhibition of cell proliferation was evaluated by MTT assay.Dose of median (IC50 )of drugs (alone or in combination)and the combination index (CI)were calculated using the median-effect equation and the combination index equation of Chou-Talalay.Flow cytometry was used to detect the cell cycle distribution.The expression of mitochondrial respiratory membrane protein complex in AGS cells was analyzed with Western blot.Results DHA and 5-FU alone or in combination could markedly suppress the proliferation of AGS in significantly time-dependent and dose-dependent manners (P <0.05).IC50 values with DHA or 5-FU administered for 24 h and 48 h were 5 1.60 μg/mL (DHA:24 h),34.82 μg/mL (DHA:48 h),45.90 μg/mL (5-FU:24 h),and 1 6.86 μg/mL (5-FU:48 h), respectively.DHA remarkably strengthened the inhibitory effect of 5-FU and decreased IC50 of 5-FU by 3.56 -2.1 5 folds.The combination of DHA and 5-FU showed synergism.Flow cytometry showed that AGS cells treated with DHA and 5-FU were arrested in G0/G1 phase and the proportion of AGS cells in G0/G1 phase increased compared with that in the control group,DHA group and 5-FU group,while the proportion of the cells in S phase decreased significantly (P < 0.05 ).Western blot showed after treatment with DHA and 5-FU for 48 h,the expression of mitochondrial respiratory membrane protein complex was significantly decreased compared with control group,DHA group and 5-FU group (P <0.05).Conclusion DHA could act synergistically with 5-FU in inhibiting the growth of gastric carcinoma cells,and meanwhile decrease the dose of 5-FU.The mechanism may be associated with cell cycle arrest in G0/G1 phase and interference in the energy metabolism of AGS cells due to inhibition of the expression of mitochondrial oxidative respiratory chain complexes by the two compounds.

Objective To evaluate the clinical effect of a novel computer-assisted navigation technique for intraoperative correction of femoral rotation deformity in diaphyseal fractures.Methods From November 2015 to November 2016,a navigation system (BrainLAB,Germany) was used in antegrade intramedullary nailing for 13 patients with femoral shaft fracture to intraoperatively restore the normal length and rotation of the fractured femur.They were 11 men and 2 women,with an average age of 38.2 years.The iujury affected the left side in 5 cases and the right side in 8.According to the Winquist Classification,there were 6 cases of type Ⅰ,3 ones of type Ⅱ,3 ones of type m,and one of type Ⅳ.This navigation system allowed the surgeons to detect and set the femoral anteversion (FAV) and length of the injured leg at the desired angle and length of the healthy contralateral femur,precisely matching the contralateral limb and restoring the normal length and rotation of the fractured femur.All the patients underwent postoperative CT scan of bilateral femora for measurement of the lengths and rotations which were conpared with the intraoperative values obtained with the navigation system.Results Additional operative time required for computerized navigation averaged 42.8 min (from 35 to 55 min).The mean length difference between the treated and untreated femora was 4.2 nnn (from 2 to 9 mm).The FAVs obtained from intraoperative navigation and postoperative CT scan were 34.0° ± 8.4° and 33.5° ± 8.3° in the healthy side and 31.2° ± 8.5° and 32.8° ± 9.0° in the injured side,showing no significant differences either between the 2 sides or between intraoperation and postoperation (P ＞ 0.05).The mean rotational difference between the 2 extremities were 4.8° ± 1.6° for the navigation and 3.8° ± 1.9° for the CT scan,showing an insignificant difference (P ＞ 0.05).All the incisions healed well with no intraoperative or postoperative complications.Conclusions This novel navigation technique may serve as a reliable tool to accurately correct the rotational malalignment of femoral shaft fractures intraoperatively,but care should be taken in every step of the navigation procedure to reduce complications.

Objective To research the relation of early growth response gene-1(EGR-1) and NF-κB in human T-cell leukemia virus type 1(HTLV-1) Tax protein positive cells. Methods RT-PCR was used to amplify the aimed segments EGR-1 cDNA which was then inserted into an eukaryotic expression plasmid pcDNA3.0 to construct pcDNA3.0-EGR-1. The constructed plasmid was transfected into TaxN and TaxP cells by Tfx-50-mediated transfer method, the expression levels of EGR-1, p65 and Tax mRNA in transfected cells were assay by RT-PCR after 48 h post-transfection, the proteins of EGR-1 and p65 were detected by Western blot after 48 h post-transfection too. The constructed plasmid and pNF-κB-luc reporter gene plasmid was co-transfected into TaxN and TaxP cells by Tfx-50-mediated transfer method, and the activity of luciferase was assay after 48 h post-transfection. Results The results showed that the eukaryotic expression plasmid pcDNA3.0-EGR-1 was successfully constructed. The mRNA and protein expression of EGR-1 could be promoted significantly by Tax. EGR-1 can promote the mRNA and protein expressions of p65 in TaxP cells, the activity of NF-κB was up-regulated by EGR-1 too. Conclusion EGR-1 maybe involve in adult T-cell leukemia(ATL) by increasing the activation of NF-κB.

ABSTRACT:Objective To investigate the effects of ω-3 polyunsaturated fatty acids on the proliferation of human gastric cancer cell line AGS and the possible mechanisms.Methods Human gastric cancer line AGS and human microvascular epithelial cell HMEC-1 were treated with different concentrations of docosdhexaenoic acid (DHA)and eicosapentaenoic acid (EPA).The inhibition of cell proliferation was evaluated by MTT assay and cell morphology.Flow cytometry was used to detect the cell cycle change.The expressions of mitochondrial respiratory membrane protein complex Ⅰ,Ⅱ and V were analyzed with Western blot.Results DHA and EPA could markedly inhibit the proliferation of AGS in significant time-dependent and concentration-dependent manners (P < 0.05 ). The inhibitory effect of DHA was stronger than that of EPA under the same concentration gradient (P <0.05).The morphological changes of cells were characterized by cell shrinkage and weak adhesion.Flow cytometry showed that AGS cells treated with DHA and EPA were arrested in G0/G1 phase and the proportion of AGS cells in G0/G1 phase increased compared with that of the control group while the proportion of the cells in S phase decreased significantly (P <0.05).Western blot showed after treatment with DHA for 24 h and 48 h,compared with control group,the expressions of mitochondrial respiratory membrane protein complex Ⅰ,Ⅱ and Ⅴ were obviously decreased.The longer effect of DHA,the lower expression of membrane protein complex (P <0.05).DHA had little effect on cell proliferation,morphology or mitochondrial membrane protein complex in HMEC-1 (P >0.05).Conclusion ω-3 PUFAs can selectively inhibit the growth and proliferation of human gastric cancer cell line AGS.These effects may be as-sociated with arresting cell cycle in G0/G1 phase and inhibiting the energy metabolism of AGS cells.

In order to construct a eukaryotic co-expression plasmid containing membrane-anchored Sjcl4FABP and Sjc26GST genes and identify their expression in vitro, Sj14 and Sj26 genes were obtained by RT-PCR with total RNA of Schistosoma japonicum adult worms as the template and cloned into eukaryotic expression plasmid pVAC to construct recombinant plasmids pVAC-Sj14 and pVAC-Sj26. Then a 23 amino-acid signal peptide of human interleukin-2 (IL-2) upstream Sj14 or Sj26 gene and a membrane-anchored sequence containing 32 amino-acids of carboxyl-terminal of human placental alkaline phosphatase (PLAP) downstream were amplified by PCR as the template of plasmid pVAC-Sj14 or pVAC-Sj26 only to get two gene fragments including Sj14 gene and Sj26 gene. The two modified genes were altogether cloned into a eukaryotic co-expression plasmid pIRES, resulting in another new recombinant plasmid pIRES-Sj26-Sj14. The expression of Sj14 and Sj26 genes was detected by RT-PCR and indirect immunofluorescent assays (IFA) when the plasmid pIRES-Sj26-Sj14 was transfected into eukaryotic Hela cells. Restriction enzyme analysis, PCR and sequencing results revealed that the recombinant plasmids pVAC-Sj14, pVAC-Sj26 and plRES-Sj26-Sj14 were successfully constructed and the expression of modified Sj14 and Sj26 genes could be detected by RT-PCR and IFA. A bivalent membrane-anchored DNA vaccine encoding Sj14 and Sj26 genes was acquired and expressed proteins were proved to be mostly anchored in cellular membranes.

To construct a recombinant plasmid Pet23a-M, the gene encoding severe acute respiratory syndrome (SARS) coronavirus membrane protein was amplified by RT-PCR and cloned into the expression plasmid Pet23a. Results of restriction endonuclease analysis, PCR detection and DNA sequencing analysis revealed that the cloned DNA sequence was the same as that reported. The recombinants were transformed into Escherichia coli (E. Coli) BL21 (DE3) and induced by Isopropyl-beta-D-thiogalactopyranoside (IPTG). The expression of 27 kD (1 kD=0.992 1 ku) protein was detected by SDS-PAGE and pured by metal chelated chromatography. Results of Western-blot showed that this expressed protein could react with antibodies in sera of SARS patients during convalescence. This provided the basis for the further study on SARS virus vaccine and diagnostic agents.
Cloning, Molecular ; Escherichia coli/genetics ; Escherichia coli/metabolism ; Membrane Proteins/*biosynthesis ; Membrane Proteins/genetics ; Membrane Proteins/isolation & purification ; Plasmids/biosynthesis ; Plasmids/genetics ; Recombinant Proteins/biosynthesis ; Recombinant Proteins/genetics ; Recombinant Proteins/isolation & purification ; Reverse Transcriptase Polymerase Chain Reaction ; SARS Virus/chemistry ; SARS Virus/*genetics ; Viral Vaccines/biosynthesis

The objective of this study was to determine the concentrations of calcium,phosphorus and magnesium in blood and compare the differences in seven areas of Heilongjiang province,and then estimate calcuim-phosphorus metabolism of beef cattle in seven beef cattle farms to provi detheoretical foundation for the prevention of calcium phosphorus metabolism diseases of beef cattle.Seven beef cattle farms of Daqing,Shuangyashan,Jiusan and Mudanjiang in Heilongjiang province were selected as the survey sites,which were recorded as group A (both grazing and stall-feeding in Shuangyashan),group B (stall-feeding mode in Shuangyashan),group C (stall-feeding mode in Daqing),group D (grazing mode in Jiusan),group E (both grazing and stall-feeding in Daqing),group F (both grazing and stall-feeding in Daqing) and group G (stall-feeding mode in Mudanjiang).Then the concentrations of Ca,Mg,P,free fatty acid (NEFA),glucose (Glc) and β-hydroxybutyric acid (BHBA) in the blood were compared to estimate the calcuim-phosphorus metabolic states.Results showed that the concentrations of calcium,magnesium and phosphorus in the plasma of 65 beef cattle in seven survey sites were within the normal range,and there was no significant difference in calcium concentration among seven sites.The P contents in group C and G were significantly higher those that in group A and B(P＜0.01),which in group C was significantly higher than those in group D,E and F (P＜0.01),which in group G was significantly higher than those in group D,E and F(P＜0.01).NEFA content in group B was significantly higher than that in group D (P＜0.05),and there was no difference among other groups.The concentration of Glc in group A was significantly higher than that in group E (P＜0.05),which in group B was significantly higher than those in group A and D,and was very significantly higher than those in group E,F and G (P＜0.01),which in group C was very significantly higher than those in group A,D,E,F and G (P＜0.01),which in group C was significantly higher than those in group A,D,E,F and G (P＜0.01),which in group D was significantly higher than that in group E (P＜0.05),and which in group F was significantly higher than that in group E (P＜0.05).The concentration of BHBA in group C,D and E were significantly higher than that in group A (P＜0.01),which in group C was significantly higher than that in group B (P＜0.05),which in group D and E was significantly higher than that in group B (P＜0.01),which in group C was significantly higher than that in group G (P＜0.01),which in group D was significantly higher than those in group F and G (P＜0.01),and in group E was highly significantly higher than that in group F and G (P＜0.01).Overall,there were not calcuim-phosphorus metabolic disorders within the seven beef cattle farms which were selected,but it is also necessary to strengthen feeding management and health care to prevent the occurrence of nutrition and metabolic diseases.

In order to construct a eukaryotic co-expression plasmid containing membrane-anchored Sjcl4FABP and Sjc26GST genes and identify their expression in vitro, Sj14 and Sj26 genes were obtained by RT-PCR with total RNA of Schistosoma japonicum adult worms as the template and cloned into eukaryotic expression plasmid pVAC to construct recombinant plasmids pVAC-Sj14 and pVAC-Sj26. Then a 23 amino-acid signal peptide of human interleukin-2 (IL-2) upstream Sj14 or Sj26 gene and a membrane-anchored sequence containing 32 amino-acids of carboxyl-terminal of human placental alkaline phosphatase (PLAP) downstream were amplified by PCR as the template of plasmid pVAC-Sjl4 or pVAC-Sj26 only to get two gene fragments including Sjl4 gene and Sj26 gene. The two modified genes were altogether cloned into a eukaryotic co-expression plasmid pIRES,resulting in another new recombinant plasmid pIRES-Sj26-Sj 14. The expression of Sj14 and Sj26genes was detected by RT-PCR and indirect immunofluorescent assays (IFA) when the plasmid pIRES-Sj26-Sj 14 was transfected into eukaryotic Hela cells. Restriction enzyme analysis, PCR and sequencing results revealed that the recombinant plasmids pVAC-Sj14, pVAC-Sj26 and pIRES-Sj26-Sj14 were successfully constructed and the expression of modified Sj 14 and Sj26 genes could be detected by RT-PCR and IFA. A bivalent membrane-anchored DNA vaccine encoding Sj14 and Sj26 genes was acquired and expressed proteins were proved to be mostly anchored in cellular membranes.