In order to enhance the specificity of TNF-α monoclonal antibody to inflamed site, a bispecific antibody BsDb that targets TNF-α and the extra-domain B (ED-B) of fibronectin (FN) was constructed by covalently linking the anti-TNF-α single chain Fv antibody (TNF-scFv) and the anti-ED-B scFv L19 via a flexible peptide linker deriving from human serum albumin (HSA). ED-B is an antigen specifically expressed at the inflamed site. BsDb is expressed in E. coli, identified by immunoblot, and purified with affinity chromatography. This was followed by further examination of its bioactivities and pharmacokinetics. We demonstrated that BsDb retained the immunoreactivity of its original antibodies as it could simultaneously bind to TNF-α and ED-B and neutralize the biological action of TNF-α. In the collagen-induced arthritis mice model, BsDb selectively accumulate in the inflamed joint with a maximal uptake of (12.2 ± 1.50)% ID/g in a single inflamed paw and retain in the inflamed paw for at least 72 h. In contrast, BsDb showed a short serum half-life of (0.50 ± 0.05) h and a rapid clearance from normal tissues. The findings reported herein indicate that BsDb has good specificity to the inflamed site and low toxicity to normal tissues. BsDb is therefore likely to have greater clinical applications in the treatment of rheumatoid arthritis and other autoimmune diseases. This laid a stable basis for its preclinical study.
Animals ; Antibodies, Bispecific ; chemistry ; Antibodies, Monoclonal ; chemistry ; Arthritis, Experimental ; Escherichia coli ; Fibronectins ; chemistry ; Half-Life ; Humans ; Mice ; Single-Chain Antibodies ; chemistry ; Tumor Necrosis Factor-alpha ; chemistry

In order to demonstrate the taxonomic position of paramyxovirus Tianjin strain and explore its mechanism of pathogenesis. Bioinformatics methods were used to analyze the deduced amino acid sequences of NP, P, M, and L protein of Tianjin strain. Phylogenetic analysis based on NP, P, M, and L protein sequences demonstrated that Tianjin strain belonged to the genus Respirovirus, in the subfamily Paramyxovirinae and most likely a new genotype of Sendai virus. Sequence similarities comparisons indicated that Tianjin strain P protein was poorly conserved, sharing only 78.7%~91.9% amino acid identity with 6 known Sendai viruses, while L protein was the most conserved, having 96.0%~98.0% amino acid identity with other Sendai viruses. Multiple-sequence alignments of Tianjin strain NP, P, M, and L protein with those of 6 known Sendai viruses showed that Tianjin strain possessed a lot of unique amino acid substitutions in protein sequences, 15 in NP, 29 in P, 6 in M, and 29 in L. The presence of these unique amino acid substitutions suggests that Tianjin strain maybe has a significant difference in host or pathological characteristics from the known Sendai viruses.

OBJECTIVETo study the expressions and the significance of interleukin-8 (IL-8) and cyclooxygenase-2 (COX-2) in the remnant stomach.

METHODSFifty-eight patients with gastrectomy were examined by upper gastrointestinal endoscopy. Two biopsy specimens were obtained from the stoma and the upper corpus gastric mucosa in the remnant stomach. mRNA was extracted from biopsy specimens to measure the IL-8 and COX-2 gene mRNA levels by real-time PCR method.

RESULTSIL-8 and COX-2 levels were higher in stoma than in corpus, IL-8 levels in BI anastomosis were significantly higher in stoma than in corpus (P< 0.05). In Hp-negative patients, IL-8 and COX-2 levels in stoma were significantly higher in BII anastomosis than in BI anastomosis (P < 0.05). In Hp-positive patients, IL-8 and COX-2 levels in stoma showed no significant differences between BII anastomosis and BI anastomosis. In corpus, IL-8 and COX-2 levels in Hp-positive patients were significantly higher than those in Hp-negative patients, (P < 0.05), including in BI anastomosis and in BII anastomosis.

CONCLUSIONSThe risk of the secondary stomach carcinogenesis in stoma after distal gastrectomy is higher than that in corpus; The types of anastomosis may influence the risk for the secondary stomach carcinogenesis in the remnant stomach mucosa.

Adult ; Aged ; Aged, 80 and over ; Female ; Gastric Mucosa ; metabolism ; microbiology ; Gastric Stump ; surgery ; Gastroenterostomy ; adverse effects ; methods ; Helicobacter Infections ; Helicobacter pylori ; Humans ; Interleukin-8 ; biosynthesis ; genetics ; Male ; Middle Aged ; Prostaglandin-Endoperoxide Synthases ; biosynthesis ; genetics ; RNA, Messenger ; biosynthesis ; Stomach Neoplasms ; etiology

OBJECTIVETo study the expression and significance of interleukin-8 (IL-8), cyclooxygenase-2 (COX-2) and trefoil family factor 1 (TFF1) in the remnant stomach mucosa.

METHODSPatients after gastrectomy were examined by upper gastrointestinal endoscopy. Biopsy specimens were obtained from stoma and the greater curvature of the upper corpus to be assessed for Hp (by H.E. and Giemsa staining) and conduct real-time semi-quantitative PCR. mRNA was extracted from the biopsy specimens to determine the IL-8, COX-2 and TFF1 gene mRNA levels by real-time PCR method.

RESULTSIn the stoma, COX-2 level in Hp-positive patients was significantly higher than that in Hp-negative patients, but the difference of IL-8 levels between them was not significant. In the corpus, IL-8 and COX-2 levels in Hp-positive patients were significantly higher than those in Hp-negative patients. In Hp-negative patients, IL-8 and COX-2 levels in the stoma were significantly higher in B II anastomosis than in B I anastomosis cases; COX-2 level in the stoma was significantly higher in B II anastomosis than in B I anastomosis cases, but the difference of IL-8 levels between them was not significant. TFF1 level in the remnant stomach mucosa showed no significant difference between Hp-positive and Hp-negative patients.

CONCLUSIONHp infection and bile reflux are important risk factors for the secondary stomach carcinogenesis. Expression of IL-8 and COX-2 in the remnant stomach mucosa is related to the risk of secondary stomach carcinogenesis. The relationship between the TFF1 expression and secondary stomach carcinogenesis in the remnant stomach mucosa is still unclear and should further be studied.

Adult ; Aged ; Aged, 80 and over ; Cyclooxygenase 2 ; biosynthesis ; genetics ; Female ; Gastrectomy ; Gastric Mucosa ; metabolism ; Gastric Stump ; Helicobacter Infections ; metabolism ; Helicobacter pylori ; Humans ; Interleukin-8 ; biosynthesis ; genetics ; Male ; Middle Aged ; RNA, Messenger ; biosynthesis ; genetics ; Risk Factors ; Stomach Neoplasms ; metabolism ; microbiology ; surgery ; Trefoil Factor-1 ; Tumor Suppressor Proteins ; biosynthesis ; genetics

BACKGROUNDGreen tea is an important source of flavonoids in human diets and epidemiological data correlate green tea consumption with a reduced cancer risk. Given its complicated properties at effective concentrations, we put epigallocatechin-3-gallate (EGCG) that previously reported on its anti-proliferative activities against several cancer cell lines on our research agenda to further examine the mechanism of its chemopreventive potential.

METHODSRNA interference (RNAi) expression vector pSilencer 3.1-H1 was used to construct recombinant nuclear factor erythroid 2 related factor 2 (Nrf2)-targeting RNAi plasmids. EGCG (5 microg/ml) was added into the culture fluid of cells before and after transfection. RT-PCR and Western blotting were used to detect the expression of uridine 5'-diphosphate-glucuronosyltransferase (UGT) 1A in cells. Forty male BALB/c mice were assigned to four groups: a normal unexposed control and three groups treated with varying doses of EGCG. Four weeks later, the mice were sacrificed, and their colon tissues were subjected to mRNA and protein expression of Nrf2 and UGT1A via RT-PCR and Western blotting analysis.

RESULTSEGCG up-regulated the expression of Nrf2 and increased the level of UGT1A in cells. The blockade of Nrf2 activity via RNA intervention largely attenuated the induction of UGT1A expression by EGCG. In mice, the mRNA and protein levels of Nrf2 and UGT1A detected by RT-PCR and Western blotting increased (both P < 0.05 compared with the control). This increase in Nrf2 expression also had a positive correlation with an increased UGT1A expression.

CONCLUSIONSEGCG mediated its effect in part by inducing the NRF2 signaling pathway and increasing UGT1A expression. Both in vitro and in vivo studies demonstrated the role of NRF2 and UGT1A expression in the potential use of EGCG as a possible chemopreventive agent and supported further study of EGCG for cancer treatment.

Animals ; Anticarcinogenic Agents ; pharmacology ; therapeutic use ; Blotting, Western ; Caco-2 Cells ; Catechin ; analogs & derivatives ; pharmacology ; therapeutic use ; Colonic Neoplasms ; drug therapy ; metabolism ; Gene Expression Regulation ; Gene Expression Regulation, Neoplastic ; Glucuronosyltransferase ; genetics ; metabolism ; Humans ; Male ; Mice ; Mice, Inbred BALB C ; NF-E2-Related Factor 2 ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction

Sargentodoxae Caulis was prepared from the stems of Sargentodoxa cuneata. Twenty compounds from the the stems of S. cuneata collected in Huangshan Mountain, Anhui province, were isolated and purified by column chromatography on macroporous resin (HPD100), silica gel, Sephadex LH-20 and semi-preparative HPLC. Their structures were elucidated on the basis of physico-chemical properties and spectral data analyses as (7R,8S)-3,3 '-5-trimethoxy-4,9-dihydroxy-4',7-expoxy-5',8-lignan-7'-en-9'-oic acid 4-O-beta-D-glucopyranoside(1), 1-O-(vanillic acid) -6-O-vanilloyl-beta-D-glucopyranoside(2), 4-hydroxyphenylethyl-6-O-coumaroyl-beta-D-glucopyranoside(3), citrusin B(4), cinnamoside(5), (-) -isolariciresinol 4'-O-beta-D-glucopyranoside (6), (-) -isolariciresinol 4-O-beta-D-glucopyranoside (7), 1-O-(vanillic acid) -6-(3", 5"-dimethoxy-galloyl) -beta-D-glucopyranoside (8), 4-hydroxyphenyl-ethyl-6-O-(E) -caffeoyl-beta-D-glucopyranoside (9), (-)-syringaresinol 4'-O-beta-D-glucopyranoside (10), (-)-syringaresinol di-O-beta-D-glucopyranoside (11), aegineoside (12), calceolarioside B (13), 4-hydroxy-3-methoxy-acetophenone-4-O-alpha-L-rhamnopyranosyl-(1 --> 6)-beta-D-glucopyranoside (14), 4-hydroxy-3-methoxy-acetophenone-4-O-beta-D-apiofuranosyl-(1 --> 6) -beta-D-glucopyranoside (15), (-) -epicatechin (16), salidroside (17), 3,4-dihydroxy-phenyl ethyl-beta-D-glucopyranoside (18), chlorogenic acid (19) and protocatechuic acid (20). Compound 1 was a new compound and compounds 2-7 were isolated from this plant for the first time.
Lignans ; isolation & purification ; Plant Stems ; chemistry ; Ranunculaceae ; chemistry

OBJECTIVETo test the reliability and validity of two mental workload assessment scales, i.e. subjective workload assessment technique (SWAT) and NASA task load index (NASA-TLX).

METHODSOne thousand two hundred and sixty-eight mental workers were sampled from various kinds of occupations, such as scientific research, education, administration and medicine, etc, with randomized cluster sampling. The re-test reliability, split-half reliability, Cronbach's alpha coefficient and correlation coefficients between item score and total score were adopted to test the reliability. The test of validity included structure validity.

RESULTSThe re-test reliability coefficients of these two scales and their items were ranged from 0.516 to 0.753 (P < 0.01), indicating the two scales had good re-test reliability; the split-half reliability of SWAT was 0.645, and its Cronbach's alpha coefficient was more than 0.80, all the correlation coefficients between its items score and total score were more than 0.70; as for NASA-TLX, both the split-half reliability and Cronbach's alpha coefficient were more than 0.80, the correlation coefficients between its items score and total score were all more than 0.60 (P < 0.01) except the item of performance. Both scales had good inner consistency. The Pearson correlation coefficient between the two scales was 0.492 (P < 0.01), implying the results of the two scales had good consistency. Factor analysis showed that the two scales had good structure validity.

CONCLUSIONBoth SWAT and NASA-TLX have good reliability and validity and may be used as a valid tool to assess mental workload in China after being revised properly.

Adolescent ; Adult ; Aged ; Female ; Humans ; Male ; Mental Competency ; Middle Aged ; Occupational Health ; Reproducibility of Results ; Sampling Studies ; Self-Assessment ; Surveys and Questionnaires ; standards ; Task Performance and Analysis ; Workload

The methods to determine the total phenols, total saponins, and marker constituents salidroside, chlorogenic acid and 3, 4-dihydroxy-phenylethyl-β-D-glucopyranoside in the samples of Sargentodoxae Caulis were established to provide the evidence for the improvement and revision of the quality standard of the crude material recorded in the Chinese Pharmacopoeia (2015 Edition). The content of total phenols was determined by ultraviolet spectrophotometry, using gallic acid as a reference substance. The content of total saponins was determined by ultraviolet spectrophotometry, using 3-O-[β-D-xylopyranosyl-(1-2)-O-β-D-glucuronopyranosyl]-28-O-[β-D-glucopyranosyl] asiatic acid as a reference substance. The contents of salidroside, chlorogenic acid and 3,4-dihydroxy-phenylethyl-β-D-glucopyranoside were detected by HPLC. The linear ranges were 1.01-7.04 mg x L(-1) for total phenols, 37.7-201 μg for total saponins, 0.025 8-1.55 μg for salidroside, 0.076 2-5.44 μg for chlorogenic acid, and 0.064 9-3.47 μg for 3,4-dihydroxy-phenylethyl-βP-D-glucopyranoside, respectively. Their average recoveries were 99.12%, 99.11% 105.5%, 99.08%, and 101.6%, respectively. The contents of total phenols and total saponins were 3. 04% -11. 9% and 0. 87% -3. 63%. The contents of salidroside, chlorogenic acid and 3,4-dihydroxy-phenylethyl-β-D-glucopyranoside fluctuated from 0.018% to 0. 572%, from 0.041% to 1.75% and from 0.035% to 1.32%. The established methods were reproducible, and they could be used for the quality control of Sargentodoxae Caulis. The present investigation suggested that total phenols, salidroside, and chlorogenic acid should be recorded in the quality standard of Sargentodoxae Caulis and their contents should not be less than 6.8% for total phenols, 0.040% for salidroside, and 0.21% for chlorogenic acid.
China ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; analysis ; Magnoliopsida ; chemistry ; Phenol ; analysis ; Plant Stems ; chemistry ; Saponins ; analysis ; Triterpenes ; analysis

Paramyxovirus Tianjin strain is the high-pathogenic virus to primate and might also cause human lower respiratory tract infection. To determine the genome structure, variation features and phylogenetic position, the complete nucleotide sequence of paramyxovirus Tianjin strain was analyzed. The homology comparison and phylogenetic analysis of the nucleotide and the deduced amino acid sequences among paramyxovirus Tianjin strain and the 28 strains in seven genera and the 7 unclassified viruses of Paramyxoviridae were performed. The results suggested that Tianjin strain is a member of the Respirovirus genus in the Paramyxovirinae, Paramyxoviridae and has the closest relationship to Sendai virus. Its genome length and composition are similar to the previously published Sendai virus except one extra glutamic acid residue increasing at the C terminus of Large protein due to the genomic RNA mutation at position A15240C. 440 unique nucleotide variations of Tianjin strain lead to 110 amino acid residue changes, making it differed from any other Sendai viruses. The phylogenetic analysis reveals paramyxovirus Tianjin strain doesn't belong to any of the three known evolution lineages of Sendai viruses and locates at a separate evolution branch. The obvious distinctions of genome nucleotide sequence, host tropism and pathogenicity suggest that paramyxovirus Tianjin strain might represent a novel genotype of Sendai virus.
Base Sequence ; Evolution, Molecular ; Genome, Viral ; Paramyxoviridae ; classification ; genetics ; Phylogeny ; Polymerase Chain Reaction ; RNA, Viral ; chemistry ; Sendai virus ; genetics

OBJECTIVETo determine the minimum number of lymph nodes that should be examined for the UICC/AJCC TNM classification of gastric cancer.

METHODSThe clinical and pathological data of four hundred and thirty-six patients underwent curative resection for gastric cancer were analyzed by Chi-square and Student-Newman-Keuls test.

RESULTSThe pN0 patients with 1 to 4, 5 to 9 examined nodes showed significantly lower survival rate than those with 10 to 14, 15 or more examined nodes (P < 0.05), and the patients with 10 to 14 examined nodes had as good a prognosis as those with 15 or more examined nodes. In the pN1, pN2 categories, the patients with 1 to 4, 5 to 9 and 10 to 14 examined nodes tended toward significantly lower survival rates than those with 15 or more examined nodes (P < 0.05). Among the patients who were classified as stage II, the survival rate of those with 10 to 19 examined nodes was significantly lower than that with 20 or more examined nodes. Among the patients classified as stage III, those with 5 to 9, 10 to 19 and 20 to 29 examined nodes had significantly lower survival rates than those with 30 or more examined nodes (P < 0.05).

CONCLUSIONSThe number of lymph nodes examined has significant prognostic impact within each pN category of gastric carcinoma. The minimum number of lymph nodes to examine in order to determine pN0 classification can be reduced from 15 to 10. For tumor with lymph node metastasis, the minimum number should be 15. In stage II, 20 or more nodes should be examined, and in stage III and IV 30 or more.

Adult ; Aged ; Female ; Humans ; Lymph Nodes ; pathology ; Male ; Middle Aged ; Neoplasm Staging ; methods ; Prognosis ; Stomach Neoplasms ; classification ; mortality ; pathology ; Survival Rate