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Objective:To investigate postoperative pathological diagnosis upgrade of high-grade cervical squamous intraepithelial lesions (HSIL) in postmenopausal women and its influencing factors.Methods:Clinicopathologic data of 378 post-menopausal women with HSIL who underwent cervical conization or total hysterectomy in Shanxi Bethune Hospital between January 2017 and December 2021 were retrospectively analyzed. According to whether the pathological diagnosis was upgraded after operation, they were divided into upgraded group and non-upgraded group. The clinicopathological characteristics of both groups were compared. Multivariate logistic regression was used to analyze the influencing factors of postoperative pathological upgrade.Results:Among 387 patients, 28 patients (7.2%) were postoperatively upgraded to cervical cancer. Compared with the non-upgraded group, the proportions of the following indexes in the upgraded group were higher [the proportion of HSIL detected by cervical thinprep cytologic test (TCT): 57.1% (16/28) vs. 44.6% (160/359); the proportion of HSIL detected by colposcopic impression: 89.3% (25/28) vs. 59.3% (213/359); the proportion of glandular involvement: 46.4% (13/28) vs. 24.0% (86/359); the number of lesion involvement ≥ 2: 82.1% (23/28) vs. 59.6% (214/359); the proportion of positive endocervical curettage (ECC): 64.3% (18/28) vs. 46.0% (165/359)]; and the differences were statistically significant (all P < 0.05). There were no statistically significant differences in the proportions of patients stratified by menopausal duration, colporrhagia, gravidity frequency, reproductive frequency, human papillomavirus (HPV) 16/18 infection and multiple HPV infection (all P > 0.05). Multivariate logistic analysis found that colposcopic impression of HSIL ( OR = 6.195, 95% CI 1.432-26.804), glandular involvement ( OR = 2.468, 95% CI 1.050-5.801), and ECC positive ( OR = 3.477,95% CI 1.028-11.764) were independent risk factors for postoperatively upgraded to cancer for postmenopausal HSIL patients in women (all P < 0.05). Conclusion:For post-menopausal women, patients with colposcopic impression of HSIL, glandular involvement and ECC positive should be alert to the risk of postoperatively pathological upgrade.
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Objective To study the distribution of connexin 43 (Cx43) in cervical cancer HeLa cells, and to verify the localization of Cx43 in mitochondria. Methods HeLa cells were segregated into cytoplasm, cell nucleus, mitochondria and supernatant after segregation by using the method of homogenate and centrifuge. Immunoelectron microscope was used to observe the morphology of mitochondria and the localization as well as the distribution of Cx43 in HeLa cells. Voltage-dependent anion channel 1 (VDAC1) was used to confirm the localization of mitochondria. Immunofluorescence was used to costain HeLa cells with Cx43 and mitochondrial marker VDAC1 to verify mitochondria localization of Cx43 in cervical cancer HeLa cells. Then Western blot was used to quantify the expression of Cx43 in fractions (cytoplasmic fraction,nuclear, mitochondria and post mitochondrial supernatant). Mitochondrial markers including VDAC1 and cytochrome c oxidaseⅣ(COXⅣ) were used to confirm mitochondria. Plasma membrane marker (LHR) was used to confirm plasma membrane. Results Immunoelectron microscope confirmed that the normal mitochondria or cystic swollen one could be seen in the complete HeLa cells and the detached HeLa cells mitochondria, with the presence of Cx43 and VDAC1 in detached mitochondria. Immunofluorescence showed Cx43 colocalized with VDAC1. There was a significant difference in the Cx43 expressions of the subcellular structure in the HeLa cells [cytoplasm (1.23±0.11), cell nucleus (0.39±0.09), mitochondria (3.67±0.59), supernatant after segregation (0.16±0.06); F =84.17, P ＜0.05]. It also showed that the relative amount of Cx43 in mitochondria was enriched. Conclusions Cx43 is enriched in mitochondria in cervical cancer HeLa cells. Therefore, Cx43 in mitochondria might be a potential target in diagnosis, therapy and prognosis of cervical cancer.
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Objective To investigate the inhibitory effect of anti interleukin(IL)-8 monoclonal antibodies on the growth and metastasis of cervical cancer. Methods Involved cervical cells included CaSki cells with high expression of IL-8 and SiHa cell lines with IL-8 plasmid transfected (pcDNA3.1-IL-8-SiHa). Cervical cancer animal model was established on nude mice. Boyden method was used in vitro study to observe the effects of anti IL-8 antibodies on the chemotaxis of high-expressed IL-8 cervical cancer cells. The effect of anti IL-8 antibodies on the growth of cervical cancer cells and nude mice transplantation tumor was observed by the experiment in vivo through reverse transcription-polymerase chain reaction (RT-PCR), enzyme linked immunosorbent assay (ELISA), TUNEL method. Cell line (CaSki and pcDNA3.1-IL-8-SiHa) modeled nude mice were divided into 5 groups with 5 animals in each group. The blank control group (group Ⅰ) was given the equal volume of phosphate buffer solution (PBS). Negative control group (group Ⅱ) was injected with IgG at the same volume of IgG. Treatment group (group Ⅲ) was injected with anti IL-8 antibodies at dose of 100 μg for once and intervals for once 2 days. Treatment group (group Ⅳ) was injected with anti IL-8 antibodies at dose of 500 μg for once and intervals for once 3 days. Treatment group (group V) was injected with anti IL-8 antibodies at dose of 1 000 μg for once and intervals for once 1 week.Results Experiments in vitro showed that the cell chemotaxis ability of anti IL-8 antibody in CaSki cells and pcDNA3.1-IL-8-SiHa cells was lower than that in the blank control group(CaSki cells:F=289.6,P =0.000; pcDNA3.1-IL-8-SiHa cells:F=79.0,P=0.005).GroupⅣwas taken as the example for its best anti-tumor effect in experiments in vivo. The tumor weight in groupⅣwas lower than that in groupⅠ[CaSki cells: (0.172±0.031) g vs. (0.735± 0.015) g, P＜ 0.05; pcDNA3.1-IL-8-SiHa cells: (0.400±0.029) g vs. (1.430±0.199) g, P＜ 0.05]. The tumor volume in groupⅣwas less than that in groupⅠ[CaSki cells:(0.049±0.028)cm3vs.(0.214±0.016) cm3,P＜0.05;pcDNA3.1-IL-8-SiHa cells:(0.063±0.022)cm3vs.(0.600±0.072)cm3,P＜0.05].The tumor growth curve also showed that tumor growth was slow, and the time of tumor formation as well as survival time was prolonged in anti IL-8 antibody treated group. The expression of mRNA in IL-8 in group IV was lower than that in group Ⅰ (CaSki cells: 0.58±0.06 vs. 1.15±0.13, P＜ 0.05; pcDNA3.1-IL-8-SiHa cells: 0.69±0.08 vs. 1.16±0.13,P＜0.05).The protein expression of IL-8 in groupⅣwas lower than that in groupⅠ(CaSki cells:126±29 vs.411±112,P＜0.05;pcDNA3.1-IL-8-SiHa cells:134±47 vs.327±69,P＜0.05).Apoptotic index in groupⅣwas higher than that in groupⅠ(CaSki cells:81.8±3.0 vs.26.0±5.6,P＜0.05;pcDNA3.1-IL-8-SiHa cells: 84.4±3.6 vs. 32.0±4.9, P＜0.05). Conclusion Anti IL-8 antibody can inhibit cell migration of human cervical cancer in vitro, inhibit growth and metastasis of transplantation tumor in vivo, and promote apoptosis and necrosis with a dose-dependent way in vivo.
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Objective To investigate the value of the multiple short tandem repeat (STR)amplification by fluorescence labeling polymerase chain reaction (PCR) combined with fusion gene bcr-abl mRNA expression for quantitative determination of chimerism and qualitative detection of bcr-abl transcripts,and to evaluate the status of engraftment and predict the outcome of allogeneic hematopoietic stem cell transplantation (allo-HSCT). Methods 5 relapse patients with CML after alIo-HSCT were dynamically investigated. Quantitative analysis of donor chimerism was performed by multiplex PCR amplification of STR markers and capillary electrophoresis with fluorescence detection, qualitative detection of bcr-abl transcripts was performed by RT-PCR. Results The donors alleles appeared in all of 5 patients on day 28 post transplant, and bcr-abl expression was negative. But 5 patients had unstable mixed ehimerism. (DC: 0～80.4 %) at the different time points after aIIo-HSCT and bcr-abl was positive. One of them kept eontinuely the mixed chimerism in the relapse of disease, and died after one year, and the other changed from MC to CC by intervention of clinical treatment. Reduction of donor chimerism were detected prior to the occurrence of graft rejection and disease relapse, while bcr-abl gene expression was positive. Conclusion The results of STR-PCR in the range of its sensitivity fully correspond with bcr-abl tests in patients with CML. The combination of STR-PCR with RT-PCR provides a highly sensitive and valuable tool for engraftment evaluation, graft rejection, relapse and predicting GVHD. Furthermore it can provide a basis for early intervention of clinical treatment, and can identify these patients at high risk with molecular or cytogenetic relapse after allo-HSCT.
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Objective To observe the neuroprotective effect of Copolymer 1 ( Cop 1) in rats with elevated intraocular pressure (IOP), and explore the clinical application of Cop 1 in the prevention of optic nerve atrophy induced by glaucoma. Methods Thirty adult Wistar rats were rendered for elevation of IOP in both eyes by Shareef-Sharma method. At the third week of IOP elevation, a mixture (1:1) of Cop 1 (100 μg) emulsified in complete Freund's adjuvant (CFA) was injected intracutaneously at two hind footpads in 15 rats (Cop 1 model group), and the other 15 rats were given the same dose of PBS emulsified in CFA (model control group) . Retinal ganglial cell (RGC) retrograde labeling was performed with Fluorogold 6 d later, and retina was taken after 24 h to compared the density of RGC between two groups. Another normal rats were served as blank controls. Results It was found that lymphocytes were accumulated on the histological slices of retina and optic nerve in Cop 1 model group. The rates of RGC loss of Cop 1 model group and model control group were 10. 24% ± 3.75% and 29.00% ±6.92%, respectively. The RGC density in Cop 1 model group was significantly lower than that in model control group (P <0. 05), while there was no significant difference in that between Cop 1 model group and blank control group (P >0.05). Conclusion Intracutaneous injection of Cop 1 can reduce the damage of RGC induced by elevated IOP.
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<p><b>BACKGROUND</b>To profile the expression patterns of 60 lung cancer related genes in human bronchial epithelial cell (BEP2D) and alpha-particle induced malignantly transformed cell (R15Hp35T-2).</p><p><b>METHODS</b>Sixty lung cancer related cDNAs were micro-arrayed onto the microscope slides using Cartesian PixSys5500 cDNA Microarray machine. Total RNA from BEP2D cell and R15Hp35T-2 cell was extracted and labeled by fluorescent dye. The labeled probe was then hybridized with the cDNA.</p><p><b>RESULTS</b>Compared with the BEP2D cell, 27 genes up-regulated and 7 down-regulated in the R15Hp35T-2 cell. The expression abundance of most tumor suppressor genes were similar in the two kinds of cells, however, most oncogenes and growth factor genes were overexpressed in R15Hp35T-2 cell.</p><p><b>CONCLUSIONS</b>In malignantly transformed human bronchial epithelial cell model induced by alpha-particle, some oncogenes and growth factor genes may promote the malignant transformation together.</p>