Objective To investigate the relationship between the expression of β-catenin and drug-resistance mechanism of choriocarcinoma according to the expression of β-catenin in JEG-3 cells (human choriocarcinoma cell line) and drug resistant JEG-3/VP16 cells.Methods The mRNA and protein expressions of β-catenin were analyzed with reverse transcription-polymerase chain reaction(RT-PCR) and Western blotting.Flow cytometry was used to determine the percentages of β-catenin-positive cells in the two choriocarcinoma cell lines.Results Both drug resistant choriocarcinoma cells and drag sensitive cells were found to express β-catenin; but the expression of β-catenin mRNA (1.43 ±0.24) and protein(1.49 ±0.17)in drug resistant choriocarcinoma cells was found much higher than that in drug sensitive cells(0.65 ±0.14,0.66 ±0.16,P ＜0.01).And according to detect by flow cytometry,we found the number of β-catenin-positive cells in JEG-3/VP16 cells [(40.13 ±5.17) ％] was much more than that in JEG-3 cells [(13.15 ± 1.48) ％,P ＜ 0.01].Conclusions β-catenin was highly expressed in the drug resistant choriocarcinoma cell line (JEG-3/VP16).It indicates β-catenin might be involved in the drug resistance mechanism of choriocareinoma.

Objective: To investigate the relationship between hemoglobin levels and prognosis of cerebral infarction in elderly patients aged 75 years and over. Methods: A retrospective analysis of 238 elderly patients (≥75 years old) with cerebral infarction admitted into our hospital from January 2016 to June 2018 was performed. The age, gender, serum creatinine and risk factors for stroke (hypertension, diabetes, dyslipidemia, homocysteine, atrial fibrillation, smoking, drinking), coronary heart disease, previous stroke history, tumor history, National Institutes of Health Stroke Scale (NIHSS) score, hemoglobin, hematocrit and other basic data were recorded. The patients were divided into the group with good prognosis (mRS score ≤ 2) and the group with poor prognosis (mRS score > 2). The relationship between hemoglobin levels and the prognosis of cerebral infarction in elderly patients were analyzed. Results: After 6 months of follow-up, 124 (52.1%, 124/238) stroke patients had a good prognosis, and 114 (47.9%, 114/238) patients had a poor prognosis, including 21 deaths. Anemia at admission (OR=2.433, 95%CI: 1.213-4.591, P=0.011), new-onset anemia after hospitalization (OR=2.615, 95%CI: 1.333-6.521, P=0.043) and the low level of minimum hemoglobin during hospitalization (OR=0.847, 95%CI: 0.671-0.971, P=0.038) were independent risk factors for poor prognosis in elderly stroke patients aged ≥75 years. New-onset anemia after hospitalization (OR=1.015, 95%CI: 1.002-1.027, P=0.022), the low level of minimum hemoglobin during hospitalization (OR=0.801, 95%CI: 0.654-0.981, P=0.027), the decrement of hemoglobin ≥20 g/L (OR=1.342, 95%CI: 1.011-1.763, P=0.000) at reexamination were independent risk factors for the mortality in elderly patients with cerebral infarction. Conclusions Anemia at admission, new-onset anemia after hospitalization, and the low level of minimum hemoglobin during hospitalization are independent predictors for poor prognosis in cerebral infarction patients aged ≥75 years. The decrement of hemoglobin ≥20 g/L after admission is an independent predictor for high mortality in elderly patients with cerebral infarction.

This study was purposed to construct and prepare the recombinant adenovirus vector carrying human thioredoxin (hTRX) and enhanced green fluorescence protein (EGFP), and transfect it into HEK293 cells, so as to lay a foundation for further gene therapy. The PCR-amplified products of hTRX with a pair of primers containing Not I and EcoR V restriction sites were subcloned into shuttle plasmid pDC316-mCMV. HEK293 cells were co-transfected with the constructed recombinant shuttle plasmid pDC316-hTRX-EGFP and large adenovirus-helper plasmid pBHGlox (delta) E1, 3Cre in mediation of liposome. The obtained replication-defective recombinant adenovirus pAd-hTRX-EGFP was co-transfected in HEK293 cells, purified by CsCl gradient centrifugation, counted for virus particles and determined for titer. The recombinant adenovirus was identified by PCR. The HEK293 cells were then transfected with adenoviruses and assayed by flow cytometry. The expression of hTRX was confirmed by Western blot. The results showed that according to PCR and restriction endonuclease assay, the target gene was inserted into recombinant adenovirus vector successfully. The sequence of fusion gene was the same as that of designed fragments. The titer of the purified recombinant adenovirus pAd-hTRX-EGFP was 5.558×10(10) pfu/ml. A transfection efficiency of 92.25% could be achieved at MOI = 100. Western blot further confirmed that hTRX was efficiently expressed in HEK293 cells. It is concluded that recombinant adenovirus vector containing hTRX has been constructed successfully and obtained highly efficient virus that can express efficiently in HEK293 cells, which laid a foundation for further investigation.
Adenoviridae ; genetics ; Genetic Vectors ; Green Fluorescent Proteins ; genetics ; HEK293 Cells ; Humans ; Plasmids ; Recombinant Proteins ; genetics ; Thioredoxins ; genetics ; Transfection

Objective To evaluate the physiological and anatomic basis,indications,surgical skills, prevention of ureter injury and clinic outcomes of using high uterosacral ligament suspension(HUS)for correction of advanced uterine prolapse by the vaginal route.Methods Fifty women with advanced uterine prolapse underwent transvaginal HUS after vaginal hysterectomy with reconstruction of pubocervical and rectovaginal fascia to correct their uterine prolapse between June 2003 and September 2007.The average age of the women was 60.1 years.The mean follow-up period was 24 months(range 4-51 months).The degree of pelvic organ prolapse preoperatively and anatomic outcomes postoperatively were assessed with pelvic organ prolapse quantification system(POP-Q).Results The remnants of the uterosacral ligaments were clearly identified and palpated posterior and medial to the ischial spines by traction with a 24 cm long Allis clamp and used for successful vaginal vault suspension and reconstruction in all 50 consecutive advanced uterine prolapse patients.The ureter injury was avoided by complete knowledge of the ureter's course from the cervix/apex toward its insertion in the sacral region and how far outside of the uterosacral ligament,by uteri palpation and by suturing purposefully placed"deep"dorsally and posteriorly toward the sacrum,as well as by cystoscopy examination of the spillage of urine from both ureters.Mean POP-Q point C improved from 1.5 to-7.5 cm with a median follow-up of 24 months.If the successful HUS was defined as point C≤stage I prolapse,both the objective and subjective cure rates were as high as 100% with a maximum follow-up of 51 months.None of the 50 patients had repeat operation for recurrence of prolapse.There was no major intra-or postoperative complications,such as ureter and other pelvic organ injury.Conclusion HUS with fascial reconstruction seems to be a safe,minimal traumatic,tolerable and highly successful procedure for vaginal repair of advanced uterine prolapse.Because of the use of native tissue as suspension site HUS is more physiologic and cost effective.

Patent foramen ovale (PFO) is a common congenital heart disease that can cause cryptogenic stroke through paradoxical embolization.For patients with PFO combined with cryptogenic stroke,whether anticoagulant therapy is superior to antiplatelet therapy in the prevention of recurrent stroke? And whether PFO closure can significantly reduce the risk of stroke recurrence compared with medical therapy alone? All those raised the clinical problems to be solved urgently.The advances in treatment of cryptogenic stroke associated with PFO are herewith summarized in present paper by reviewing randomized trials,meta-analyses and the guidelines or expert consensus about PFO and cryptogenic stroke.

Objective:To investigate the influence of metastasis suppressor gene KAI1 on the proliferation,invasion and metastasis of endometrial carcinoma cell line AN3CA and HEC-1-B.Methods:The KAI1 cDNA was transfected into human endometrial carcinoma cells AN3CA and HEC-1-B via Lipofectamine 2000.The expression of KAI1 protein was ex- amined by Western blotting and flow cytometry before and after transfection.The proliferation ability of AN3CA and HEC- 1-B cells was observed by MTT assay and anchorage-independent growth assay.The changes of cell invasive ability were studied by transwell assays.Results:Stable expression of KAI1 protein was observed in AN3CA and HEC-1-B cells and on their surface after transfection with pcDNA3-KAI1 plasmid.Cells transfected with blank plasmid formed more colonies and had a larger size,with the colony forming rates being(54.2?3.1)% for AN3CA cells and(52.7?4.3)% for HEC- 1- B cells;the doubling time of AN3CA and HEC-1-B cells were 21.3 h and 20.1 h,respectively.Cells transfected with pcDNA3-KAI1 formed less colonies and had a smaller size,with the colony forming rates being(37.4?5.1)% for AN3CA cells and(32.1?3.7)% for HEC-1-B cells;the doubling time of AN3CA and HEC-1-B cells were 43.7h and 45.2 h,respectively.The cell proliferation abilities and colony-forming ability were significantly different between the two groups(P

Objective　To evaluate significance of squamous cell carcinoma associated antigen (SCC Ag) and carcinoembrynoic antigen (CEA) and carbohydrate antigen 15-3 (CA15-3) in early diagnosis of lung squamous cell carcinoma. Methods　Lung squamous cell carcinoma was induced with methylcholanthrene (MCA) in iodized oil in Wistar rats. Ninty-one rats were used for this experiment, and the rats were sacrificed on the 20th, 40th, 50th, 60th, 70th and 80th days respectively. The stage of the cancer development including atypical hyperplasia, carcinoma in situ, early invasive carcinoma was diagnosed histopathologically. SCC Ag, CEA and CA15-3 in the serum of different conceration stage rats were measured by MEIA technique. Results　Serum levels of SCC Ag, CEA and CA15-3 of the model rats in the atypical hyperplasia and carcinoma in situ had no significant difference. The level of SCC Ag increased predominantly at the early invasive carcinoma(P＜0.01), but the other two markers had no change. Conclusion　SCC Ag level in serum may be of considerable importance in the early diagnosis of lung squamous cell carcinoma.

Background:Leukocyte telomere has been shown to be related to insulin resistance-related diseases,such as type 2 diabetes mellitus (T2DM) and non-alcoholic fatty liver disease (NAFLD).This cross-sectional study investigated the association of leukocyte telomere length (LTL) with NAFLD in T2DM patients.Methods:Clinical features were collected and LTL was measured by Southern blot-based terminal restriction fragment length analysis in 120 T2DM patients without NAFLD and 120 age-matched T2DM patients with NAFLD.NAFLD was clinically defined by manifestations of ultrasonography.The correlation between LTL and clinical and biochemical parameters were analyzed by Pearson correlation or Spearman correlation analysis.Factors for NAFLD in T2DM patients were identified using multiple logistic regressions.Results:LTL in T2DM patients with NAFLD were significantly longer than those without NAFLD (6400.2 ± 71.8 base pairs [bp] vs.6023.7 ± 49.5 bp,P ＜ 0.001),especially when diabetes duration was less than 2 years.Meanwhile,the trend of shorter LTL was associated with the increased diabetes duration in T2DM patient with NAFLD,but not in T2DM patients without NAFLD.Finally,LTL (odds ratio [OR]:1.001,95％ confidence interval [CI]:1.000-1.002,P =0.001),as well as body mass index (OR:1.314,95％ CI:1.169-1.477,P ＜ 0.001) and triglycerides (OR:1.984,95％ CI:1.432-2.747,P ＜ 0.001),had a significant association with NAFLD status in T2DM patients.Conclusions:T2DM patients with NAFLD had a significantly longer LTL than those without NAFLD.The longer LTL was especially evident in the early stage of T2DM,indicating that longer LTL may be used as a biomarker for NAFLD in T2DM patients.

OBJECTIVETo investigate the mechanism of clinical haemorrhage in an inherited coagulation factor VII (FVII) deficiency and tissue factor abnormality pedigree.

METHODSAll exons, exon-intron boundaries and the 3', 5' untranslated sequences of FVII and tissue factor (TF) genes were amplified by PCR and sequenced directly. Any mutation identified by direct sequencing was confirmed by reverse sequencing. FVII cDNA of the proband was synthesized with random primers and amplified by nest PCR.

RESULTS55C-->T heterozygous mutation located in promoter of FVII gene was identified in the proband. The heterozygous mutation was derived from his mother. Tracing the other pedigree members found that his sister had the same heterozygous mutation and the others had wild-type FVII genes. A 9363 C-->T (Arg131Trp) heterozygous polymorphism in TF gene, which was 2.63% frequency of T allele polymorphism, was found in all of the pedigree members.

CONCLUSIONIt was the first report that the -55C-->T heterozygous mutation in FVII gene and the Arg131Trp heterozygous polymorphism in TF gene explained the clinical symptom of the proband.

Adult ; DNA Mutational Analysis ; Factor VII ; genetics ; Factor VII Deficiency ; genetics ; Heterozygote ; Humans ; Male ; Pedigree ; Polymorphism, Genetic ; Thromboplastin ; genetics

Adult ; Amyloidosis ; complications ; Factor X Deficiency ; complications ; Humans ; Liver Diseases ; complications ; Male