By using a yeast two-hybrid system, a yeast two-hybrid bait vector was constructed and identified for screening of the HPV18 E6-interacting proteins, and its effects on the growth of yeast cells and the activation of reporter genes were investigated. Total mRNA extracted from Hela cells was reversely transcribed into cDNA. Fragment of HPV18 E6 cDNA was amplified using RT-PCR and directly ligated to the pGBKT7 vector. The recombinant plasmid was confirmed by restriction endonuclease analysis and DNA sequencing. The recombinant pGBKT7-HPV18 E6 plasmid and empty pGBKT7 vector were transformed into the yeast cell AH109, respectively. After they were cultured respectively in YPDA liquid medium and nutrition-deficient culture medium, their toxicity and transcriptional activation were tested by both the phenotype assay and the color assay. The bait plasmid HPV18 E6 was successfully obtained. After being cultured in YPDA liquid medium for 16h, the A (600 nm) values of two yeast fluids were 0.98+/-0.03 and 0.99+/-0.02, respectively. The recombinant pGBKT7-HPV18 E6 plasmid and empty pGBKT7 vector could grow to white colonies on SD/-Trp/X-alpha-gal plates, while no colony could survive on SD/-His/-Trp/X-alpha-gal, SD/-Ade/-Trp/X-alpha-gal plates, indicating that the bait plasmid pGBKT7-HPV18 E6 was constructed successfully and expressed correctly, and could not activate the transcription of reporter gene alone. The yeast two-hybrid GAL4 system 3 can be utilized to find HPV18 E6 interacting proteins.

Small cell carcinoma of cervix (SCCC) is a rare disease with highly aggressive behaviour and is pathologically hard to diagnose. In this study, the clinicopathological features, diagnosis, treatment and prognosis of the condition were examined. Clinical records and follow-up data of 7 cases of SCCC were retrospectively studied. Our results showed that five non-recurrent cases initially presented irregular vaginal bleeding or increased apocenosis of varying degrees. Pathological examination revealed that the stroma was diffusely infiltrated with small monomorphous cells ranging from round to oval shape. Three cases were immunohistochemically confirmed. One case was accompanied with squamous cell cancer. Of the 7 cases, one case was classified as stage I b1, two stage I b2, one stage IIa, one stage IIb, and one stage IIIb. On the basis of their stages of condition, one subject with stage III b underwent chemotherapy, and one with stage Ib2 received extensive hysterectomy plus pelvic lymphadenectomy, while the other 5 cases were treated by extensive hysterectomy and pelvic lymphadenectomy in combination with pre- and/or post-operative adjuvant chemotherapy and radiotherapy. Of the 7 patients, 4 had relapse-free survival of 14, 14, 16 and 28 months respectively. It is concluded that SCCC is an aggressive tumor with propensity for early pelvis lymph node metastases. Early-stage patients should be treated by extensive hysterectomy and pelvic lymphadenectomy in combination with pre- and/or post-operative adjuvant chemotherapy and radiotherapy.

OBJECTIVETo analyze the correlation between skeletal muscle mass and age among check-up adults.

METHODSThe study objects were those who aged 18-87 years old and came to a city hospital of Qiqihar for health examination from December, 2013 to September, 2014, excepted those with coronary heart disease, infectious disease, endocrine system disease, hypertension, anemia, cancer, liver disease, kidney disease and those carrying with heart pacemaker. A total of 1 836 respondents were finally enrolled into analysis. Appendicular Skeletal Muscle (ASM) was measured by a Body Composition Analyzer, and relative skeletal muscle index (RSMI) was calculated. The relationship among ASM, RSMI and age was assessed by linear regression analysis. The difference of height, weight, BMI, waist-hip-ratio (WHR), total muscle mass and percentage of body fat between genders were tested by t-test. The difference of ASM and total skeletal muscle mass between genders and among age groups was tested by multi-factor variance analysis. The difference of the muscle decline between genders was compared by Chi-square test.

RESULTSThe total muscle mass in males was (52.22 ± 6.65) kg, which was significantly higher than that in females ((38.05 ± 4.39) kg) (t=28.20, P<0.001). ASM in 18-29, 30-39, 40-49, 50-59, 60-69, 70-87 years was (24.64 ± 3.23), (24.00 ± 3.12), (24.35 ± 3.03), (23.33 ± 2.97), (22.54 ± 2.91) and (21.40 ± 3.36) kg (F=16.12, P<0.001) in males, respectively, and (16.48 ± 3.14), (16.72 ± 1.93), (16.75 ± 1.93), (16.84 ± 2.28), (16.52 ± 2.35) and(14.70 ± 2.37)kg (F=4.38, P=0.001) in females, respectively. ASM in males ((23.72 ± 3.16) kg) was higher than that in females ((16.65 ± 2.25) kg) (t=55.97, P<0.001). There was a negative correlation between age and ASM in males after 50 years old, the regression equation was y=28.31-0.09x (P<0.001). While a negative correlation between age and ASM in females occurred after 60 years old, the regression equation was y=27.69-0.18x (P<0.001). The prevalence of low ASM was 16.85% (124/736) in females, which was significantly higher than that in males (8.73%, 96/1 100) (χ(2)=27.57, P<0.001).

CONCLUSIONA negative correlation was found between age and ASM in males after 50 years old and in females after 60 years old. The prevalence of low RSMI in females was significantly higher than that in males.

Adipose Tissue ; Adolescent ; Adult ; Aged ; Aged, 80 and over ; Body Composition ; Body Mass Index ; Body Weight ; China ; Female ; Humans ; Male ; Middle Aged ; Muscle, Skeletal ; physiology ; Prevalence ; Young Adult

Small cell carcinoma of cervix (SCCC) is a rare disease with highly aggressive behaviour and is pathologically hard to diagnose. In this study, the clinicopathological features, diagnosis,treatment and prognosis of the condition were examined. Clinical records and follow-up data of 7 cases of SCCC were retrospectively studied. Our results showed that five non-recurrent cases initially presented irregular vaginal bleeding or increased apocenosis of varying degrees. Pathological examination revealed that the stroma was diffusely infiltrated with small monomorphous cells ranging from round to oval shape. Three cases were immunohistochemically confirmed. One case was accompanied with squamous cell cancer. Of the 7 cases, one case was classified as stage Ⅰ bl, two stage Ⅰ b2,one stage Ⅱ a, one stage Ⅱ b, and one stage Ⅲb. On the basis of their stages of condition, one subject with stage Ⅲ b underwent chemotherapy, and one with stage Ib2 received extensive hysterectomy plus pelvic lymphadenectomy, while the other 5 cases were treated by extensive hysterectomy and pelvic lymphadenectomy in combination with pre- and/or post-operative adjuvant chemotherapy and radiotherapy. Of the 7 patients, 4 had relapse-free survival of 14, 14, 16 and 28 months respectively.It is concluded that SCCC is an aggressive tumor with propensity for early pelvis lymph node metastases. Early-stage patients should be treated by extensive hysterectomy and pelvic lymphadenectomy in combination with pre- and/or post-operative adjuvant chemotherapy and radiotherapy.

Objective:To investigate the value of the virtual monoenergetic image (VMI) obtained by a new dual-layer detector spectral CT combined with metal artifact reduction algorithms(O-MAR) in reduction of different types of artifacts caused by 125I seeds implantation and in improvement of the post-operative CT image quality. Methods:This was a cross-sectional study. Thirty-five patients who underwent dual-layer detector spectral CT scanning of the chest and abdomen after 125I seeds implantation were retrospectively included at the First Affiliated Hospital of Zhengzhou University from March to September 2022. The spectral data were collected and reconstructed into conventional CT image (CI), VMI image (50-150 keV, 20 keV/level), CI+O-MAR image, and VMI+O-MAR image (50-150 keV, 20 keV/level). The artifacts′ removal effects and image quality improvement in each group were evaluated. Two slices with the strongest artifacts were selected for analysis for each patient, resulting in a total of 70 slices. Objective indicators including artifact index (AI), signal-to-noise ratio (SNR) and contrast-to-noise ratio (CNR) of soft tissue regions affected by artifacts were measured and calculated. Subjective indicators including the overcorrected artifacts and new artifacts, the different forms of artifacts, the diagnosis of artifacts, and the image quality were assessed. One-way analysis of variance was used for comparisons among multiple groups. Paired t test was used to compare the quantitative indicators between the combined O-MAR group and the non-O-MAR group. Kappa statistics was used to evaluate the consistency between observers. Results:In high/low-density artifacts (ROI H/L), the AI values in all groups showed decrease with increasing VMI keV. In artifact-affected tissue (ROI T), SNR of the CI/VMI (70-150 keV)+O-MAR group were significantly higher than those of the CI/VMI group ( P<0.05), CNR of the CI/VMI(50-150 keV)+O-MAR group were significantly higher than those of the CI/VMI group ( P<0.05). Both overcorrection and new artifacts mainly presented in VMI 50 keV and VMI 70 keV groups; Compared with VMI (50-70 keV) group, significantly less numbers of overcorrection and new artifacts were found in VMI (50-70 keV)+O-MAR group ( P<0.05); regarding the comparison of artifact types, with the VMI keV increasing, the number of a-type banded artifacts gradually decreased on images with high-density artifacts, reaching a minimum of 3 in the VMI 150 keV+O-MAR group; while the number of e-type artifacts with little or no artifacts increased, with the highest number of 23 in the VMI 150 keV+O-MAR group. The total number of high-density artifacts in each type decreased with increasing VMI keV. As VMI keV increased, the diagnostic and image quality scores of high-density artifacts in each group were significantly higher than those of low-density artifacts in the VMI+O-MAR group ( P<0.05). Conclusions:VMI combined with O-MAR can significantly improve the objective and subjective image quality of follow-up CT imaging after 125I seed implantation, enhancing lesion visibility and diagnostic confidence. Additionally, VMI+O-MAR showed more pronounced correction effect on high-density artifacts.

By using a yeast two-hybrid system,a yeast two-hybrid bait vector was constructed and identified for screening of the HPV18 E6-interacting proteins,and its effects on the growth of yeast cells and the activation of reporter genes were investigated.Total mRNA extracted from Hela cells was reversely transcribed into cDNA.Fragment of HPV18 E6 cDNA was amplified using RT-PCR and directly ligated to the pGBKT7 vector.The recombinant plasmid was confirmed by restriction endonuclease analysis and DNA sequencing.The recombinant pGBKT7-HPV18 E6 plasmid and empty pGBKT7 vector were transformed into the yeast cell AH109,respectively.After they were cultured respectively in YPDA liquid medium and nutrition-deficient culture medium,their toxicity and transcriptional activation were tested by both the phenotype assay and the color assay.The bait plasmid HPV18 E6 was successfully obtained.After being cultured in YPDA liquid medium for 16h,the A600 nm values of two yeast fluids were 0.98±0.03 and 0.99±0.02,respectively.The recombinant pGBKT7-HPV18 E6 plasmid and empty pGBKT7 vector could grow to white colonies on SD/-Trp/X-α-gal plates,while no colony could survive on SD/-His/-Trp/X-α-gal,SD/-Ade/-Trp/X-α-gal plates,indicating that the bait plasmid pGBKT7-HPV18 E6 was constructed successfully and expressed correctly,and could not activate the transcription of reporter gene alone.The yeast two-hybrid GAL4 system 3 can be utilized to find HPV18 E6 interacting proteins.

The aim of this study was to produce recombinant porcine interferon gamma (rPoIFN-γ) by Chinese hamster ovarian (CHO) cells expression system and to analyze its antiviral activity. Firstly, we constructed the recombinant eukaryotic expression plasmid pcDNA3.1-PoIFN-γ and transfected into suspension cultured CHO cells for secretory expression of rPoIFN-γ. The rPoIFN-γ was purified by affinity chromatography and identified with SDS-PAGE and Western blotting. Subsequently, the cytotoxicity of rPoIFN-γ was analyzed by CCK-8 test, and the antiviral activity of rPoIFN-γ was evaluated using standard procedures in VSV/PK-15 (virus/cell) test system. Finally the anti-Seneca virus A (SVA) of rPoIFN-γ activity and the induction of interferon-stimulated genes (ISGs) and cytokines were also analyzed. The results showed that rPoIFN-γ could successfully expressed in the supernatant of CHO cells. CCK-8 assays indicated that rPoIFN-γ did not show cytotoxicity on IBRS-2 cells. The biological activity of rPoIFN-γ was 5.59×10⁷ U/mg in VSV/PK-15 system. Moreover, rPoIFN-γ could induced the expression of ISGs and cytokines, and significantly inhibited the replication of SVA. In conclusion, the high activity of rPoIFN-γ was successfully prepared by CHO cells expression system, which showed strong antiviral activity on SVA. This study may facilitate the investigation of rPoIFN-γ function and the development of novel genetically engineered antiviral drugs.
Swine ; Animals ; Cricetinae ; Interferon-gamma/pharmacology* ; Cricetulus ; CHO Cells ; Sincalide ; Recombinant Proteins/pharmacology* ; Antiviral Agents/pharmacology*