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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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Jiujing Tu (Illustration of Moxibustion), excavated from Mo Kao Grotto at Dunhuang, is one of the earliest existing monographs on moxibustion. The medical masters from different schools have focused on this book because it is different from the existing ancient medical works and have not been collected in the medical works of different dynasties. In this study, the literature of Jiujing Tu on five acupoints (Dachangshu, Pangguangshu, Daxiaochangshu, Nieshu and Cigong) relevant with intestinal disorders is collected. It is intended to discuss and analyze the acupoint location, main intestinal disorder, moxibustion characters, recognition on the literature of different dynasties and modern clinical applications. It is believed that the thought of strong moxibustion in the treatment of intestinal disorders advocated in Jiujing Tu has profound impact on the medical development in later generations. It deserves us to have a further digging, collection and promotion of this thought in the modern time.
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Humanos , Pontos de Acupuntura , Livros Ilustrados , História , China , História Antiga , Medicina na Literatura , Meridianos , Moxibustão , HistóriaRESUMO
Objective:To evaluate and compare electroacupunctures (EA) with different parameters and moxibustion at different temperatures influencing the activation of mast cells (MC) in Tianshu (ST 25) regions of visceral hyperalgesia model rats. Methods:Rats (except for model group) respectively accepted 1 mA or 3 mA EA or moxibustion at 43 or 4 to℃℃stimulate Tianshu (ST 25) points after randomization of the fifty visceral hyperalgesia model rats, and then were compared with that in model and normal groups. Number, degranulation numbers, degranulation rates in Tianshu (ST 25) regions MC of rats in each group were observed using toluidine blue staining. Abdominal withdrawl reflex (AWR) score was used to evaluate the rat visceral hyperalgesia reactions. Results:Compared with the normal group and the model group, MC numbers (P0.05); AWR scores in 1 mA EA group under the stimulation of 0 mmHg or 80 mmHg were significantly higher than that in the normal group (P<0.01); AWR score in 3 mA EA group under the stimulation of 0 mmHg was significantly higher than that in the normal group (P<0.01), and AWR scores in 3 mA EA group under the stimulation of 20 mmHg or 80 mmHg were also higher than that in the normal group (P<0.05). AWR scores were higher in 1 mA EA group under the stimulation of 40 mmHg or 80 mmHg than that in 4℃ moxibustion group (P<0.05); AWR score was higher in 3 mA EA group under the stimulation of 40 mmHg than that in 4℃ moxibustion group (P<0.05). Conclusion:There are differences among EA of different parameters and moxibustion of different temperatures in activating on Tianshu (ST 25) regions MC of visceral hyperalgesia model rats, as well as in improving the visceral hyperalgesia reaction. The effect of 4℃ moxibustion is the most significant.
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The clinical experience of professorfor polycystic ovary syndrome (PCOS) was summarized. According to the main pathogenesis of PCOS, the tonifying kidney should be taken as essence with synchronous treatment on liver, spleen and heart, presenting staging, classification and sorting method for PCOS. In the staging method, the regulation on follicle development should be taken as treatment core to comply with the rules ofand. A four-stage method was proposed, where "regulating method" was suitable in menstrual period, "tonifying method" in follicular phase;"dredging method" in ovulatory period and "adjustment and tonifying " in luteal phase. In the classification and sorting method, attention was paid on individualized treatment, and treatment was based on fat type, thin type and non-fat type as well as childbearing. Besides, psychological counseling and life adjustment for patient was essential, and the unity of body and mind could enhance curative effect.
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Objective To investigate the impact of migration inhibitory factor (MIF) over-expression on the epithelial to mesenchymal transition in human cervical cancer SiHa cells.Methods Recombinant plasmid pEGFP-N1-MIF was transfected into SiHa cells,and then of MIF mRNA relative quantitative expression was tested by RT-PCR.The mRNA and protein expression level of E-cadherin and vimentin were detected by RT-PCR and immunocytochemistry,respectively.Results RT-PCR results showed that MIF mRNA expression quantity in experimental group was higher than that in control group cells (F =2 950.278,P ＜ 0.01).In MIF-overexpressing SiHa cells,vimentin mRNA was increased and E-cadherin mRNA was decreased determined by RT-PCR (Fvalues were 2 135.048,1 893.563,P＜ 0.01).Immunocytochemistry results showed that vimentin expression quantity in experimental group cells were higher than that in control group cells,however,E-cadherin expression quantity was lower than that in control group cells (F values were 2 348.021,1 789.421,P ＜ 0.01).Conclusions The over-expression of MIF gene can significantly up-regulate the expression of vimentin,and down-regulate the expression of E-cadherin.Consequently,MIF over-expression induces epithelial to mesenchymal transition in human cervical cancer SiHa cells.
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Objective To investigate the effects of macrophage migration inhibitory factor (MIF) overexpression on the expression of interleukin-8 (IL-8),martix metalloproteinase-9 (MMP-9) and invasion of human cervical cancer SiHa cells.Methods Chemical synthesis MIF eDNA gene,designed primer sequence including XhoI and BamHI enzyme sites,MIF gene was amplified by polymerase chain reaction (PCR),constructed eukaryotic expression vector pEGFP-N1/MIF and transfected into SiHa cells using Lipofectamine and won over-expression of MIF.The expression of MIF in supernatant fluid was detected by ELISA,the expression of MIF,IL-8,MMP-9 in both mRNA and protein levels were detected by real-time fluorescence quantitative-PCR and immunocytochemistry respectively.The effect of over-expressed MIF on migration was detected by Boyden small chamber.Results The expression of protein in supernatant fluid transfected with pEGFP-N1/MIF was significantly increased (Fgroup =8267.564,P ＜ 0.01),the expression of MIF,IL-8,MMP-9 in both mRNA and protein in SiHa cells transfected with pEGFP-N1/MIF were significantly increased (F values were 7019.619,2148.094,3303.540,1565.114,2807.300,523.466,P ＜ 0.01),and there was a positive correlation among MIF,IL-8,MMP-9 expression in both mRNA and protein (r values were 0.865,0.895,0.934,0.908,P ＜ 0.01).Invasion ability in SiHa cells transfected with pEGFP-N1/MIF was obviously increased (F=3430.898,P＜ 0.01).Conclusion The over-expression MIF gene in SiHa cells can promote cervical cancer cell invasion and metastasis of ability,which could be associated with the upregulation of IL-8 and MMP-9 expression.
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The IL-8 and MMP-7 genes participate in the carcinogenesis of many malignancies, but the role of both genes in cervical cancer is not fully elucidated. The aim of this study was to determine the frequency of IL-8 and MMP-7 gene mutations and to assess their effects on the risk of early stage cervical cancer and lymph node metastasis. The clinical stage and histological grade of cervical cancer were also studied. The peripheral blood from the patients with early stage cervical cancers and normal controls was collected and the DNA was extracted. The incidence of IL-8 and MMP-7 gene mutations was assessed by using tetra-primer amplification refractory mutation system polymerase chain reaction (ARMS PCR) and restriction fragment length polymorphism (RFLP). The data were statistically analyzed by x2 test. The results showed that: (1) The genotype frequency of IL-8 -251AT and TT was significantly higher in the cervical cancer group than in the normal control group (OR=2.290 and 2.619 respectively, P=0.001), and it was also higher in the lymphatic metastasis group than that without metastasis (OR=2.917, P=0.035); (2) The frequency of MMP-7 -181G/G genotype was significantly higher in the cervical cancer group and in the lymphatic metastasis group (P<0.05); (3) The incidence of IL-8 mutation was two times higher in IIa cervical cancer group than in Ib1 and Ib2 cervical cancer group (P=0.006). For the MMP-7 gene, there was statistically significant difference in the incidence of mutation between the Ib1, Ib2 and the IIa (P=0.000); (4) Different histological types and different grades of cervical cancer had different incidence of mutations, statistically. It was suggested that there was significant difference in the genotype of IL-8 -251TT and MMP-7 -181GG polymorphism between the cervical cancer group and the lymph node metastasis group. Moreover, individuals with IL-8 T allele or MMP-7 G allele carriers were at significantly higher risk of cervical cancer, particularly the early (IIa) and medium, poorly differentiated cervical cancer (G2+G3).
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Objective To compare the advantages and disadvantages of the amplification refractory mutation system -polymerase chain reaction (ARMS-PCR) with restriction fragment length polymorphism polymerase chain reaction(RFLP-PCR) technology, then explore its application in detection and analysis of single nucleotide polymorphism (SNP). Methods By using ARMS-PCR and RFLP-PCR the polymorphisms of the MMP-2 in cervical cancer and the normal-control group were analyzed. The research sites were-1306 C/T.The final result was verified by DNA sequencing. Results The results of amplified used ARMS-PCR showed that the genotypes of the SNP site in MMP-2 gene promoter were CC, CT, TT, and the DNA fragments were corresponding with the results of DNA sequencing. Conclusion ARMS-PCR technology has the advantages of lower cost time-saving simple operation and reliable results, while has a high value for research of SNP.
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Objective To determine the correlation between the expression of interleukin 8(IL-8) and lymph node metastases in earlier uterine squamous cervical cancer and to study the mechanism of metastases of earlier squamous cervical cancer. Methods IL-8 mRNA expression was examined with reverse transcription polymerase chain reaction(RT-PCR) and tissue microarray to determine the expression of IL-8 protein. The matrix metalloproteinase 9( MMP-9) protein and microvessel density (MVD) in earlier squamous cervical cancer were decided respectively by antibody against IL-8, MMP-9 and CD-34 associated antigen and Immunohistochemical analysis. Results The cervical cancer with lymph node metastases express IL-8, MMP-9 and MVD at significantly higher level than negtive metastatic lymph node(P =0.00,0.00,0.02). Otherwise, the express of IL-8 correlated strongly and positively with MVD of cervical cancer(r =0.251,P
