Analysis of tumor microenvironmental features to refine prognosis by T, N risk group in patients with stage III colon cancer (NCCTG N0147) (Alliance).

BACKGROUND: Tumor-infiltrating lymphocytes (TILs), tumor budding, and micropapillary architecture may influence tumor growth and metastatic potential, thereby enhancing prognostic stratification. We analyzed these features and their relative contribution to overall outcome and in low (T1-3 N1) and high (T4 and/or N2) risk groups that are used to inform the duration of adjuvant chemotherapy in patients with resected stage III colon cancers. PATIENTS AND METHODS: Among 1532 patients treated in a phase III adjuvant trial of FOLFOX-based therapy, intraepithelial TIL densities, tumor budding, and micropapillary features were analyzed and quantified in routine histopathological sections with light microscopy. Optimal cut-points were determined in association with disease-free survival (DFS) in training and validation sets. Associations or relative contributions of individual features or combined variables with DFS were determined using multivariable Cox regression models. RESULTS: TILs, tumor budding, and micropapillary features were shown to differ significantly by T, N risk groups and by mismatch repair (MMR) status. Low TILs, high budding, and their combined variable [hazard ratio = 2.07 (95% CI, 1.50% to 2.88%); Padj < 0.0001], but not micropapillary features, were each significantly associated with poorer DFS in a training data set and confirmed in a validation set. TILs were prognostic in proficient mismatch repair (pMMR) and deficient mismatch repair (dMMR) tumors; budding was prognostic only in pMMR tumors. The percentage relative contribution of budding/TILs to DFS was second only to nodal status overall, was second (24.4%) after KRAS in low-risk patients, and was the most important contributor (45.4%) in high-risk patients. CONCLUSIONS: TIL density and tumor budding were each validated as significant prognostic variables and their combined variable provided robust prognostic stratification by T, N risk groups, being the strongest predictor of DFS among high-risk stage III patients. CLINICALTRIALS. GOV IDENTIFIER: NCT00079274.

Alpha1,2-fucosyl transferase gene, the key enzyme of Lewis y synthesis, promotes Taxol resistance of ovarian carcinoma through apoptosis-related proteins.

We aimed to investigate the role of FUT1 gene in Taxol resistance and to explore its mechanism in epithelial ovarian cancer. Three ovarian cancer cell lines, ES-2, SK-OV-3 and OVCAR-3 were selected from epithelial ovarian cancer in this experiment. Western blot was used to validate the protein expression level of FUT1 and the apoptosis proteins. The expression level of the corresponding carrier was validated by RT-PCR. Transfection and isolation of stable transfectants were carried out to establish the cell line models. The different concentrations of Taxol on the inhibition of cell growth rate was measured by MTT, in which Taxol resistance profiling in ovarian cancer cells was determined by IC50 data. Flow cytometry was conducted to compare cell apoptosis ability. Caspase-3 activity and the apoptosis proteins were measured by colorimetry and western blot, respectively, to further compare the cell apoptosis ability in different groups. To demonstrate the inhibition of miR-FUT1 combined with Taxol therapy against ovarian cancer, xenograft assay was carried out for the in vivo effect. The western blot results indicate that FUT1 is expressed in all of the ovarian cancer cells with different expression level: ES-2 > SK-OV-3 > OVCAR-3. Besides, FUT1 siRNA was used in the maximum expression of FUT1 cell line ES-2, or over-expression plasmid was used in the minimum expression of FUT1 cell line OVCAR-3, to establish stable expression cell lines. After the treatment with Taxol, the inhibition rate of Taxol was obviously decreased with the established cell model above, and the IC50 level was significantly increased in the FUT1 over-expression + Taxol group (p Keywords: FUT1, Lewis y, Taxol resistance, ovarian cancer, apoptosis.

[Genetic characterization of norovirus isolated in an outbreak of gastroenteritis in Jiangsu province].

Objective: To analyze the genetic characterization of norovirus isolated in an outbreak of gastroenteritis in Jiangsu province. Methods: Extracted viral RNA from the swab samples of cases of acute gastroenteritis outbreak in Jiangsu province on December 16-27, 2016 was reversely transcribed to cDNA, and partial RNA-dependent RNA polymerase sequence and complete capsid sequence (VP1) were amplified by RT-PCR. Amplification products were sequenced for the analysis of genetic characteristics. Results: Based on sequence alignment, the variant shared a high level of identity with the strain GⅡ.g isolated in Spain and Finland (98.7%) in the RNA-dependent RNA polymerase region, and with the strain GⅡ.1 isolated in American (99.4%) in the VP1. The recombination was determined by using software Simplot, and the breakpoint of recombination was located in the ORF1/2 overlap region at position 5 106 of VP1. The result of amino acids alignment in capsid region showed that there were no mutations in the amino acids of the predicted epitopes and receptor binding site Ⅰ-Ⅲ, but a unique amino acid change was detected at position 132 (N-S). Conclusion: The norovirus isolated in the outbreak of gastroenteritis in Jiangsu province was a rare recombinant norovirus variant GⅡ.g-GⅡ.1.

3D kinematics of the equine metacarpophalangeal joint at walk and trot.

The metacarpophalangeal (MCP) joint and its supporting soft tissues are common sites of injury in athletic horses. Equine gait analysis has focused on 2D analysis in the sagittal plane and little information is available which describes 3D motions of the MCP joint and their possible role in the development of injuries. The aim was to characterize the 3D rotations of the equine MCP joint during walking and trotting. Three-dimensional trajectories of marker triads fixed rigidly to the third metacarpus and proximal phalanx of the right forelimb of healthy horses were recorded at walk (n = 4) and trot (n = 6) at 120 Hz using eight infra-red cameras. Kinematics of the MCP joint were calculated in terms of helical angles between the two segments using singular-value decomposition and spatial attitude methods. The ranges of motion were: flexion/extension: 62 +/- 7 degrees at walk, 77 +/- 5 degrees at trot; adduction/abduction: 13 +/- 7 degrees at walk, 18 +/- 7 degrees at trot; and axial rotation: 6 +/- 3 degrees at walk, 9 +/- 5 degrees at trot. Flexion/extension had a consistent pattern and amplitude in all horses and appeared to be coupled with adduction/abduction, such that stance phase extension was accompanied by abduction and swing phase flexion was accompanied by adduction. Axial rotation was small in amount and the direction varied between horses but was consistent within an individual for the two gaits.

3D kinematics of the interphalangeal joints in the forelimb of walking and trotting horses.

The objective was to measure 3D rotations of the distal (DIP) and proximal (PIP) interphalangeal joints at walk and trot. 3D trajectories of markers fixed to the proximal phalanx, middle phalanx and the hoof wall of the right forelimb of four sound horses were recorded at 120 Hz. Joint kinematics were calculated in terms of anatomically-based joint coordinate systems between the bone segments. Ranges of motion were similar at walk and trot. Values for the DIP joint were: flexion/extension: 46 +/- 3 degrees at walk, 47 +/- 4 degrees at trot; internal/external rotation: 5 +/- 1 degrees at walk, 6 +/- 3 degrees at trot; and adduction/abduction: 5 +/- 2 degrees at walk, 5 +/- 3 degrees at trot. Within each gait, kinematic profiles at the DIP joint were similar between horses with the exception of adduction/abduction during breakover, which may vary depending on the direction of hoof rotation over the toe. Knowledge of the types and amounts of motion at the DIP joint will be useful in understanding the aetiology and treatment of injuries to the soft tissues, which are being recognized more frequently through the use of sensitive imaging techniques. Ranges of motion for the PIP joint were: flexion/extension: 13 +/- 4 degrees at walk, 14 +/- 4 degrees at trot; adduction/abduction: 3 +/- 1 degrees at walk, 3 +/- 1 degrees at trot; and internal/external rotation: 3 +/- 1 degrees at walk, 4 +/- 1 degrees at trot. The PIP joint made a significant contribution to flexion/extension of the digit. During surgical arthrodesis, the angle of fusion may be important since loss of PIP joint extension in late stance is likely to be accommodated by increased extension of the DIP joint.

Head and body centre of mass movement in horses trotting on a circular path.

REASONS FOR PERFORMING STUDY: Horses are often worked along a circular trajectory but the mechanics of turning have not been reported. OBJECTIVE: To develop a methodology for kinematic analysis of horses during circling and to report preliminary findings describing movements of the total body centre of mass (COMbody). METHODS: Seventy-five skin markers, distributed over the head, neck, trunk and limbs of 5 sound horses, were tracked in a global coordinate system at 120 Hz as the horses trotted around a small circle. Based on segmental inertial data, 3D positions of the segmental centres of mass and COMbody were calculated. Movements of COMbody were measured relative to the cylindrical vertical plane, the radius of which was defined by a marker overlying the sixteenth thoracic vertebra. RESULTS: During trotting, COMbody was high at the start and end of the diagonal stance phases and low in midstance. All horses leaned to the inside of the circle, with a tilt angle of mean +/- s.d. 14.8 +/- 2.8 degrees during the entire stride. In the transverse direction, COMbody swung from side to side reaching its most inward position around the time of contact of the inside forelimb and its most outward position around the time of contact of the outside forelimb. The centre of mass of the head and neck segments (COMhead/neck) had a symmetrical, double sinusoidal pattern in the vertical direction. All horses had the same motion pattern for COMhead/neck in the transverse direction but the mean position relative to the circle line varied between horses. CONCLUSIONS: Horses lean to the inside when turning around a small circle. COMhead/neck and COMbody showed 2 vertical oscillations in each stride, whereas there was a single mediolateral excursion that was not symmetrical to the inside and outside of the circle line. POTENTIAL RELEVANCE: This study provides a first step toward understanding the mechanics of locomotion during turning, which has implications in the aetiology of injuries, the response to lungeing as a diagnostic technique and the incorporation of circles into rehabilitation exercises.

Three-dimensional analysis of patterns of skin displacement over the equine radius.

REASONS FOR PERFORMING STUDY: Surface markers are usually used to track bone movement. However, skin movement related to the bone has a large effect on the analysis of skeletal kinematics. A 2-dimensional (2D) skin displacement correction model has been successfully developed, but no 3D skin displacement model exists. OBJECTIVES: To develop a 3-dimensional (3D) skin displacement model for the equine radial segment during trot. METHODS: The 3D trajectories of 6 skin-based markers and a bone-fixed triad were captured at trot in 4 horses. Skin displacements in the bone-based coordinate system were calculated using a singular-value decomposition method. The truncated Fourier series models were developed for the skin displacements using a generalised cross-validatory spline. RESULTS: Mean + /- s.d. of peak skin displacement of the 3 markers on the proximal radius as percentage of radial length was 10.7 +/- 0.5, 4.6 +/- 1.5 and 14.5 +/- 2.9% in x, y and z direction, respectively. For the 3 markers on the distal radius, the equivalent displacements were 4.7 +/- 0.6, 1.7 +/- 0.8 and 7.3 +/- 18% in x, y and z direction, respectively. CONCLUSIONS: The 3D skin displacement model for correction of skin marker motion over the equine radius relative to the bone can be established using a truncated Fourier series, which has previously been used successfully to develop 2D models. POTENTIAL RELEVANCE: This method of determining 3D skin displacement correction needs to be extended to the entire fore- and hindlimbs to provide a more sensitive measure of kinematic analysis. Accurate descriptions of the 3D motions of the limb segments and interactions between adjacent segments at the joints are necessary for understanding of the mechanics of different gaits and the gait aberrations that manifest as lameness.

Three-dimensional carpal kinematics of trotting horses.

REASONS FOR PERFORMING STUDY: Descriptions of 3D kinematics assist in understanding joint function and dysfunction, and are an essential step toward 3D inverse dynamic analysis. OBJECTIVES: To measure 3D carpal joint motion during trotting. METHODS: Three-dimensional trajectories of bone-fixed markers on the radius and third metacarpus of the right forelimb of 3 healthy horses were recorded at 120 Hz using a 6-camera analysis system. Joint kinematics were calculated in terms of helical angles between the 2 segments using a spatial attitude method. RESULTS: All horses showed carpal extension and internal rotation of the metacarpus relative to the radius as the carpus assumed the close-packed position. In late stance, the carpus began a cycle of flexion that continued through midswing, accompanied by a small cycle of internal rotation. The direction of abduction/adduction varied between horses. The predominant rotational movement was flexion/extension, which showed a range of motion of 15 +/- 6 degrees in stance and 76 +/- 13 degrees in swing. CONCLUSIONS: Carpal motions were generally similar between horses with the exception of abduction/adduction. POTENTIAL RELEVANCE: Knowledge of carpal joint motion should assist in understanding the pathogenesis of carpal injuries. However, it seems probable that real differences exist between individuals; therefore, further investigations of the effect of conformation on carpal motion should be performed in a much larger population of horses.

Allium chemistry: identification of organosulfur compounds in ramp (Allium tricoccum) homogenates.

Supercritical fluid (SF) extracts of homogenized ramp (Allium tricoccum Ait.) were separated and characterized with liquid chromatography coupled with atmospheric pressure chemical ionization mass spectrometric identification. The profiles of SF extracts of aqueous homogenates of ramp bulbs from three different seasons and growing regions revealed that the thiosulfinates were major components. In addition, some of the cepaenes (alpha-sulfinyldisulfides) found in extracts of onion juice, as well as allyl containing cepaenes (2-propenyl l-(2-propenylsulfinyl)propyl disulfide), are present in the ramp extracts. The amount of allicin in ramp bulb homogenates ranged from approximately 10% to 50% of that found in extracts of aqueous garlic homogenates. The greater amount of the methyl 1-propenyl thiosulfinates in the ramp extracts relative to that found in the garlic extracts correlates with the flavor characteristics of ramp bulbs.

Identification and characterization of a factor which is essential for assembly of transcarboxylase.

Transcarboxylase (TC) from Propionibacterium shermanii is a biotin-containing enzyme which catalyzes the reversible transfer of a carboxyl group from methylmalonyl-CoA to pyruvate. It is composed of a central, hexameric 12S subunit with six outer, dimeric 5S subunits held in a stable 26S complex by twelve 1.3S biotinyl subunits. Each of these subunits has been cloned from the P. shermanii genome and expressed in Escherichia coli. The purified, expressed recombinant proteins are all indistinguishable from their authentic counterparts except for the recombinant 5S subunit (termed 5S WT), which does not form TC complexes or catalyze the overall transcarboxylase reaction. Circular dichroism and isoelectric focusing suggested differences existed between the authentic and E. coli-expressed 5S proteins. HPLC gel filtration was used to separate the authentic 5S dimer from additional components in the preparation. 5S dimer thus purified was unable to form TC complexes or catalyze the overall reaction, behaving identically to the recombinant 5S WT subunit. Fractions from the HPLC gel-filtration purification of authentic 5S were then added to 5S WT or 5S dimer, and one fraction was identified which catalyzed the assembly of TC complexes with either 5S preparation. This assembly activity was shown to be dependent on the concentration of this HPLC fraction. Assembly-promoting factor (APF) is heat-stable and probably a protein, on the basis of its protease susceptibility. Studies with APF and the other TC subunits demonstrate its ability to promote complex formation with 12S and 1.3S subunits. Since the APF was purified from crystals of 26S TC, we believe it to be a novel, previously unidentified subunit of transcarboxylase.

[Thin layer chromatographic studies on radix Ginseng, fructus Schisandrae and radix Salviae miltiorrhizae in yixinfumai granules].

The main constituents of Radix Ginseng, Fructus Schisandrae and Radix Salviae Miltiorrhizae in Yixinfumai Granules were studied by TLC. It was found that ginsenoside got hydrolyzed while the drugs were being boiled together. The main constituents of Fructus Schisandrae were schizandrol A and B; and those of Radix Salviae Miltiorrhizae were tanshinone and protocatechuic aldehyde.

[Determination of glaucocalyxin A in the leaves of Plectranthus japonicus (Burm.) Koidz. var. glaucocalyx (Maxim.) Koidz. by HPLC].

This paper reports the quantitative analysis of glaucocalyxin A in the leaves of Plectranthus japonicus var. glaucocalyx by HPLC. The chromatographic conditions were as follows: column: Shim-pack CLC-ODS; mobile phase: methanol-water (70:30). The average content of glaucocalyxin A in the leaves is 1.30%.

[Studies on the relation of alkaloids in semen Strychni and its processing].

This paper reports the determination of strychnine and brucine in different parts of Semen Strychni (the seeds of Strychnos nux-vomica and S. pierriana), and makes a comparison of the contents between the crude forms and processed products of the two seeds. Based on the results, the authors have made some discussion about the relationship between the alkaloid distribution in Semen Strychni and its processing.