Objective To analyze the clinical application value of endometrial collector combined with liquid based cytology and histological examination for screening of endometrial cancer.Methods One hundred and eighty-two cases high-risk people with endometrial cancer in Songgang People's Hospital of Bao'an district of Shenzhen from June 2014 to December 2015 were collected,among them 71 patients with endometrial cancer were found in the patients who were diagnosed by pathological diagnosis,and the remaining 111 were not diagnosed with endometrial cancer.The material satisfaction,sample collection time of sample collection and endometrial curettage device were comparatively analyzed and also the cervical dilatation production rate,the amount of bleeding,pain score(VAS) were analyzed.The application value of endometrial collector combined with liquid based cytology and histological examination for screening of endometrial cancer was judged according to the results obtained.Results There was no significant difference between the two methods based on satisfaction(χ2=1.953,P=0.089).A methods of cervical endometrial expansion rate(3.85% vs.93.96%) and acquisition time((16.35±2.19) min vs.(11.23±4.26) min),bleeding volume((1.64±0.63) ml vs.(15.73±5.41) ml),VAS score((0.82±0.24) points vs.(4.34±2.43) points) were significantly lower than that of curettage sample collection method(χ2=16.473,P=0.000;t=5.733,P=0.034;t=11.183,P=0.000;t=3.618,P=0.043).With curettage pathological diagnosis as the gold standard,the specificity,sensitivity and diagnostic coincidence rate of endometrial biopsy combined with liquid based cytology and histological examination for screening endometrial carcinoma were respectively 94.51%,95.05% and 94.96%.Conclusion Compared with curettage sampling method,endometrial curettage collector has features of high success rate,short time,low pain and high security and the cervical dilation rate is low.Endometrial curettage collector can be combined with liquid based cytology and histology to examine and screen endometrial cancer.This methods is worth promoting.

BACKGROUND: The central position of dendritic cells (DCs) has aroused increasing attention due to strong antigen presentation capability in anti-tumor. However, how to obtain enough functional DCs and reports regarding immunomodulator with low toxicity are few. OBJECTIVE: To investigate the effect of mycobacterium phlei F.U.36 (Utilins) on the proliferation and maturity of DCs derived from human umbilical cord blood in vitro.METHODS: The mononuclear cells were isolated from human umbilical cord blood using Ficoll-Hypaque method and cultured with Utilins, cytokine (human recombinant granulocyte/macrophage colonystimulating factor + recombinant human tumor necrosisfactor-α + recombinant human interleukin-4), or combination cytokine + Utilins, respectively. In addition, cells cultured with RPMI-1640 served as controls. Morphological features and growth of DCs were observed by an inverted microscope. Thephenotype changes of DCs, such CD1a and HLA-DR, were detected by flow cytometry at 9 days after culture. Moreover, DCs were stained by Wright-Giemsa and observed under oil lens. RESULTS AND CONCLUSION: Typical DCs with high expressions of CD1a and HLA-DR were obviously detected in all experimental groups except the control group. The positive rates of CD1a and HLA-DR in the Utilins group were higher than those in the control group, but lower than the cytokine group (P < 0.05). The HLA-DR positive rate of the combination group was higher than that of the cytokine group (P < 0.05). The results revealed that, Utilins can not only promote the proliferation of DCs derived from human umbilical cord blood in vitro but also cooperate with cytokines to induce the maturity of DCs.

BACKGROUND: The existence of minimal residual leukemia cells is the main cause for the recurrence of acute leukemia in children, and immunological biological therapy has attracted more and more attentions in the various methods from eliminating minimal residual disease. Previous studies have found that interferon α-2b can effectively inhibit the increase of tumor cells in vivo in children with neuroblastoma and malignant lymphoma, whether it can inhibit the increase of leukemia cells?OBJECTIVE: To investigate the effects of interferon α-2b in vitro on leukemia cells.DESIGN: A comparative observation taking human promyelocytic leukemia HL-60 cell line as the material.SETTING: Cell Culture Room; Immunological Laboratory; Cell Room, Institute of Pediatrics, Affiliated Hospital,Medical College of Qingdao University.MATERIALS: HL-60 cell line was provided by Shandong Institute of Basic Medical Sciences. Interferon α-2b was purchased from Megagene Company Fluorescein isothiocyanate (FTTC) rabbit-anti-rat Ig solution (CatEK001) and CD13 anti-human monoclonal antibody solution (Cat. DK013Y) were purchased from Union Stem Cell & Gene Engineering Co.,Ltd.METHODS: The experiments were carried out in the Institute of Pediatrics, Affiliated Hospital, Medical College of Qingdao University from March to September 2005. HL-60 cells culture system was established in vitro, and the oncentration was adjusted to 1×109 L-1. The cells were divided into control group and experimental group. In the experimental group, each well was added by interferon-α-2b with the terminal concentration of 5×105, 1×106, 2×106,5×106 and 1×107 U/L, respectively. In the control group, each well was added by saline of the same volume. The cells were cultured continuously for 48 hours. The morphological changes of HL-60 cells were observed using Wright's staining under light microscope; Cell apoptosis was observed using acridine orange/ethidium bromide double staining; Antigen expression and maturation and differentiation on cell membrane were observed by determining CD13 protein expression; Proliferation and activity of HL-60 were detected with methyl-thiazol-tetrazolium (MTT) assay.MAIN OUTCOME MEASURES: The occurrence of apoptosis was judged according to the uniformity and staining of HL-60 nuclear chromatin; HL-60 cell proliferation was judged according to the absorbance (A) value; The maturation of HL-60 cells was judged according to the number of positive CD13 cells.RESULTS: ① HL-60 cell apoptosis: The cells were cultured for 48 hours. When the concentration of interferon α-2b was 5×105 U/L, there were mainly early apoptotic HL-60 cells; When the concentration was 1×107 U/L, there were mainly late apoptotic cells, and the apoptotic rate was significantly higher than those in the control group (P ＜ 0.01 ).② HL-60 cell proliferation: The A values in the experimental groups treated with interferon α-2b of 2×106 U/L and 1 ×107 U/L were significantly lower than that in the control group (P ＜ 0.01). ③ Maturation of HL-60 cells: The percentages of positive CD13 cells in the experimental groups treated with interferon α-2b of 1 ×106 and 1 ×107 U/L were significantly lower than that in the control group (P ＜ 0.01).CONCLUSION: It is concluded that interferon α-2b can enhance the apoptosis, inhibit the proliferation and promote maturation and differentiation of HL-60 cells.

AIM:To explore the effect of recombinamt rat CC16 protein (rCC16) on LPS-induced expression of tumor necrosis factor-alpha (TNF-α), interleukin-6 (IL-6) and IL-8 in the rat tracheal epithelial (RTE) cells. METHODS:The RTE cells were incubated with rCC16 at concentrations of 0.5, 1.0 and 2.0 mg/L in serum-free media for 2 h prior to LPS (0.1 mg/L) treatment for further 24 h.The cells were harvested for assessing the mRNA levels of TNF-α, IL-6 and IL-8 by RT-qPCR.The cell culture supernatants were collected for analyzing the protein levels of TNF -α, IL-6 and IL-8 by ELISA.In addition, the nuclear translocation of nuclear factor-κB (NF-κB) p65 was tested by Western blot.RESULTS:rCC16 inhibited LPS-induced IL-6 and IL-8 expression at both mRNA and protein levels in the RTE cells in a concentration-dependent (0~2 mg/L) manner, as demonstrated by RT-qPCR and ELISA.However, no concentra-tion-dependent manner between the dose of rCC 16 and TNF-αexpression was observed , and rCC16 inhibited LPS-induced TNF-αexpression at lower concentration (0.5 mg/L).rCC16 concentration-dependently inhibited the effects of LPS on the level of nuclear translocation of NF-κB p65.CONCLUSION:rCC16 suppresses LPS-mediated TNF-α, IL-6 and IL-8 pro-duction through inactivation of NF-κB activity in RTE cells .

OBJECTIVE: To establish a rapid detection and genotyping method for SARS-CoV-2 Omicron BA.4/5 variants using CRISPPR-Cas12a gene editing technology. METHODS: We combined reverse transcription-polymerase chain reaction (RT-PCR) and CRISPR gene editing technology and designed a specific CRISPPR RNA (crRNA) with suboptimal protospacer adjacent motifs (PAM) for rapid detection and genotyping of SARS- CoV-2 Omicron BA.4/5 variants. The performance of this RT- PCR/ CRISPPR-Cas12a assay was evaluated using 43 clinical samples of patients infected by wild-type SARS-CoV-2 and the Alpha, Beta, Delta, Omicron BA. 1 and BA. 4/5 variants and 20 SARS- CoV- 2-negative clinical samples infected with 11 respiratory pathogens. With Sanger sequencing method as the gold standard, the specificity, sensitivity, concordance (Kappa) and area under the ROC curve (AUC) of RT-PCR/CRISPPR-Cas12a assay were calculated. RESULTS: This assay was capable of rapid and specific detection of SARS- CoV-2 Omicron BA.4/5 variant within 30 min with the lowest detection limit of 10 copies/μL, and no cross-reaction was observed in SARS-CoV-2-negative clinical samples infected with 11 common respiratory pathogens. The two Omicron BA.4/5 specific crRNAs (crRNA-1 and crRNA-2) allowed the assay to accurately distinguish Omicron BA.4/5 from BA.1 sublineage and other major SARS-CoV-2 variants of concern. For detection of SARS-CoV-2 Omicron BA.4/5 variants, the sensitivity of the established assay using crRNA-1 and crRNA-2 was 97.83% and 100% with specificity of 100% and AUC of 0.998 and 1.000, respectively, and their concordance rate with Sanger sequencing method was 92.83% and 96.41%, respectively. CONCLUSION By combining RT-PCR and CRISPPR-Cas12a gene editing technology, we successfully developed a new method for rapid detection and identification of SARS-CoV-2 Omicron BA.4/5 variants with a high sensitivity, specificity and reproducibility, which allows rapid detection and genotyping of SARS- CoV-2 variants and monitoring of the emerging variants and their dissemination.
Humans ; COVID-19 ; CRISPR-Cas Systems ; Genotype ; Reproducibility of Results ; Reverse Transcriptase Polymerase Chain Reaction ; SARS-CoV-2/genetics* ; RNA ; COVID-19 Testing