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Traditional Chinese medicine dispensing granules（TCMDGs）is the new type of decoction pieces with the development of modernization of TCM， which has received mixed opinions since its practical application. In 2021， the national departments issued Announcement on Ending the Pilot Work of TCMDGs， marking the end of the 28-year pilot work of TCMDGs， and eligible TCM enterprises can produce TCMDGs after filing. However， this does not mean that the preparation process， quality standard and efficacy research of TCMDGs have been developed and matured， on the contrary， there are still some problems that need to be solved and gradually improved. For example， in the production process， there are problems such as unclear， unified and non-standardized preparation parameters. In terms of quality control， there are some problems such as lack of producing area regulation， variety selection and processing specification. In terms of consistency evaluation with traditional decoction， there are problems such as unclear relationship between the chemical constituents and pharmacological effects of the two. Therefore， in view of some prominent problems of TCMDGs at present， this paper takes the published literature as the main data source and combines the specific requirements of the code or technical standards such as the 2020 edition of Chinese Pharmacopoeia， Publicity of the Unified Standard on the Varieties of TCMDGs， Quality Control and Standard Formulation Technical Requirements of TCMDGs. The production process of TCMDGs， the origin and variety of raw materials， the processing of decoction pieces， the quality control standard and the consistency evaluation of formula granules and traditional decoction were sorted out and visualized by literature mining， data analysis and list comparison. Based on the analysis results， the following suggestions were made. In terms of preparation process， the completeness and standardization of process parameters should be strengthened. In terms of quality evaluation， attention should be paid to the relationship between the authenticity， variety， processing and quality of medicinal materials. In the consistency evaluation of formula granules and traditional decoction， the deep difference and mechanism between TCMDGs and traditional decoction were discussed by combining structural Chinese medicine， quality marker（Q-Marker） theory and physicochemical characterization， so as to provide reference for the application and development of TCMDGs.
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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 improvement effects and mechan ism of thalidomide on Alzheimer ’s disease （AD）model of Caenorhabditis elegans . METHODS In this study ，the BR 5270 strain of C. elegans was used as AD model and BR 5271 strain as the control. The effects of thalidomide （0.5，2.0，6.0，15.0 mg/mL）on the motility of BR 5270 strains of C. elegans were studied by the basal slowing response assay ；the effects of thalidomide （0.5，2.0，6.0，15.0 mg/mL）on the survival time of BR 5270 strain of C. elegans were studied by life assay ；the effects of thalidomide （0.5，2.0，6.0 mg/mL）on learning and memory ability of BR 5270 strain of C. elegans were studied by short-term and long-term learning and memory assay. RT-PCR technology was used to study the effects of thalidomide （0.5，2.0，6.0 mg/mL）on mRNA expression of phosphatidylinositol 3-kinase（PI3K）/protein kinase B （Akt）signal pathway related genes （Age-1，Akt-1，Gsk-3）and calpain homologous gene （Clp-1）in BR 5270 strain of C. elegans . RESULTS After the intervention of thalidomide ，oscillation times of BR 5270 strain of C. elegans increased significantly within 30 s （except for 0.5 mg/mL group ），and the 10％ of maximum life span was prolonged significantly （only 0.5 mg/mL group ）；the short-term and long-term learning indexes were improved significantly （only 6.0 mg/mL group ）；mRNA expression of Age-1 and Akt-1（except for 0.5，2.0 mg/mL groups ）were increased significantly ，mRNA expression of Gsk-3（except for 0.5 mg/mL group ） and Clp-1 were decreased （P＜0.05 or P＜0.01）. CONCLUSIONS Thalidomide can ameliorate the dyskinesia of AD model of C. elegans，prolong the lifespan of this strain ，and enhance its learning and memory ability. Its mechanism of action may be related to activation of PI 3K/Akt signaling pathway and inhibition of calpain.
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With the concepts of minimal invasive surgery and precision medicine embedding deeply in modern medicine, traditional clinical treatment strategies are constantly updated due to the rapid progression of medical research. Systematic dissection of the regional lymph nodes is a key component in the surgical therapy of gynecologic malignancies, however it may result in a series of complications, such as lower extremity lymphedema, deep venous thrombosis, immune disorders and so on, which seriously impact the patients' quality of life. The necessity of systematic lymphadenectomy and the extent of lymph nodes resection have always been one of the most controversial issues in gynecologic oncology. In recent years, the extensive application of sentinel lymph node (SLN) technology has been increasingly used in the treatment of gynecologic malignancies, which may provide important guidance on this issue. This article reviews the clinical application status, detection methods, detection efficiency, and current controversies regarding SLN mapping and biopsy in cervical cancer, endometrial cancer, ovarian cancer and vulvar cancer, discussing the application value of SLN technology in gynecologic oncology.
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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 explore the relationship between aryl hydrocar-bon receptor (AhR),aryl hydrocarbon receptor nuclear translocator (ARNT),estradiol (E2),estrogen receptor (ER) and recurrent spontaneous abortion (RSA) through observing the level of serum AhR,ARNT,E2,and AhR,ARNT,ER in decidua and chorionic tissues of the patients with recurrent spontaneous abortion.Methods 64 cases of RSA patients who induced abortion at the Shanxi Dayi hospital from May 2015 to September2017 were chosen as RSA group,and 30 cases of healthy abortion women of the same period who had born full-term normal fetus were choosen as the normal group.The serum,villi and decidua of each case were collected during abortion.The level of AhR,ARNT,ER of both groupswere detected by enzyme-linked immunoscrbent assay (ELISA) method.The Serum E2 lever were detected by the method of chemical luminescence.The data of all the patients were analyzed.Results (1) The level of ARNT in peripheral blood of RSA group was significantly higher than that of normal group (P ＜ 0.05),and the levels of AhR and E2 in peripheral blood were not statistically significant between the two groups (P ＞ 0.05).The levels of AhR and ARNT in villi and decidua were significantly higher in RSA group than in normal group (P ＜ 0.05),while the expression of ER in villi and decidua was significantly lower in RSA group than in normal group (P ＜ 0.05).(2) There was no significant difference in the ratio of AhR/ARNT between the two groups (P ＞ 0.05);the ratio of AhR/ER and ARNT/ER in the villi and decidua of the RSA group was higher than that of the normal group (P ＜ 0.05).Conclusions Overexpression of AhR and ARNT and low expression of ER in villi and decidua tissues may be related to the occurrence of RSA.
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High-risk Human papilloma virus (HR-HPV) has been identified as a necessary factor for the development of precancerous lesions and invasive carcinoma of the genital tract,among which cervical cancer is the most common cancer in the female genital tract.The molecular biological mechanism of HPV leading to malignant transformation of cervix and the epidemiological information of HPV provide us with many strategies for early detection and early intervention of cervical lesions.In recent years,the incidence and mortality of cervical cancer in our country had the trend of increasing.This article reviews the research progress of HPV and cervical cancer and precancerous lesion.
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Objective To observe the effect of drug edible of herbal medicine granule on vascular endothelial function in patients with coronary heart disease(CHD).Methods A total of 200 cases featuring with cardiovascular blood stasis were chosen from patients who were diagnosed with CHD in Zhengzhou hospital of traditional Chinese medicine from January 2015 to January 2016.All cases were divided into observation group(n=98)and control group(n=102).The observation group was given Xin bao drug edible of herbal medicine granule and conventional drugs,and the control group was given conventional drugs.After 8 weeks of observation,the levels of endothelin(ET-1),nitric oxide(NO)and angiotensin Ⅱ(AngⅡ)were measured by enzyme-linked immunosorbent assay(ELISA).The serum levels of total cholesterol(TC),triglyceride(TG),low density lipoprotein(LDL),and TCM clinical symptom score were compared between the two groups.Results The level of TC of the control group was decreased after treatment(P<0.05);the levels of TC,TG and LDL in the observation group were lower than those in the control group(P<0.05).After treatment,the levels of NO,ET-1,Ang-Ⅱ and FMD in the observation group were significantly different from those before treatment(P<0.05).The levels of NO and FMD in the observation group were higher than those in the control group,and the levels of ET-1 and Ang-Ⅱ were lower than those in the control group(P<0.05).After treatment,the scores of TCM clinical symptoms in the observation group were lower than those before treatment(P<0.05).Compared with the control group,the total score of palpitations,the times of shortness of breath and clinical symptoms were lower than those in the observation group(P<0.05).Conclusion Xin bao drug edible of herbal medicine granule could relieve coronary heart disease blood stasis type of patients with vascular tension,reduce its endothelial cell damage and improve its clinical symptoms.
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OBJECTIVE:To study the lifespan effect of ethyl acetate extract from Polygonum multiflora(EPM)on caenorhab-ditis elegans,and to investigate its anti-aging effect. METHODS:EPM low-concentration,medium-concentration and high- concen-tration groups(25,37.5,50 mg/L,calculated by crude drug)and blank group(0 mg/L)were established to investigate the effects of EPM on the lifespan of caenorhabditis elegans. EPM group(37.5 mg/L)and blank group(0 mg/L)were established to perform re-productive test,acute heat stress test and acute oxidative stress test of caenorhabditis elegans. The effects of EPM on reproductive capacity and pressure stress of caenorhabditis elegans were investigated. RESULTS:The lifespan test,average lifespan of cae-norhabditis elegans in EPM low-concentration,medium-concentration and high-concentration groups were prolonged significantly, compared to blank group(P<0.05 or P<0.01),especially in EPM medium-concentration group. In reproductive test,the number of offspring of caenorhabditis elegans in EPM group on the second and third day were increased significantly,compared to blank group (P<0.05). In acute heat stress test and acute oxidative stress test,average survival time of caenorhabditis elegans in EPM group was prolonged significantly(P<0.05). CONCLUSIONS:37.5 mg/L EPM can retard the aging process of caenorhabditis ele-gans and doesn’t damage the reproductive capacity.
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High-risk Human papilloma virus (HR-HPV) has been identified as a necessary factor for the development of precancerous lesions and invasive carcinoma of the genital tract,among which cervical cancer is the most common cancer in the female genital tract.The molecular biological mechanism of HPV leading to malignant transformation of cervix and the epidemiological information of HPV provide us with many strategies for early detection and early intervention of cervical lesions.In recent years,the incidence and mortality of cervical cancer in our country had the trend of increasing.This article reviews the research progress of HPV and cervical cancer and precancerous lesion.
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Objective To investigate the effect of neoadjuvant chemotherapy on the prognosis of patients with cervical cancer.Methods A systematic review of the literature about neoadjuvant chemotherapy and radical surgery was performed with searching Pubmed,Cochrane library,Medline,and Embase database from January 1990 to January 2015.According to the inclusion and exclusion criteria,the data were extracted and analyzed by Revman 5.3 system software Meta.Results Ten controlled clinical studies were concluded,including 1 543 patients (667 in preoperative neoadjuvant chemotherapy group and 876 in radical surgery).The extracted data were comparable.Meta-analysis results displayed that significant difference existed in five-year disease-free survival,node-positive rate,vascular invasion rate,and parametrial infiltration rate.It seemed that the preoperative neoadjuvant chemotherapy was prior to the radical surgery in those items.There was no significant difference between the preoperative neoadjuvant chemotherapy and radical surgery in five year survival,intraoperative complication,the positive surgical margin rate,and five-year survival rates among different chemotherapy peers (P ＞ 0.05).Conclusions For locally advanced cervical cancer patients,neoadjuvant chemotherapy can reduce the incidence of high-risk pathology,but had no significant effect on the long-term survival of patients.
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Objective To investigate the effect of over-expressed macrophage migration inhibitory factor (MIF) on epithelial-mesenchymal transition (EMT) in human cervical carcinoma SiHa cells.Methods Recombinant eukaryotic expression plasmid liposome enhanced transfection of green fluorescent protein gene (pEGFP-N1)-MIF was constructed and then transfected into human cervical cancer SiHa cells.Experimental cells were classified into three groups (SiHa-pEGFP-N1-MIF,SiHa-pEGFP-N1,and SiHa).Western blot was used to detect the expression of MIF protein,and the expressions of EMT-related markers such as E-cadherin and vimentin in SiHa cells were determined before and after transfection.Results The eukaryotic expression vector pEGFP-N1-MIF significantly increased the expression of MIF protein in SiHa cells (P ＜ 0.05),and after overexpression of MIF gene in SiHa cells,the expression of E-cadherin protein in SiHa-pEGFP-N1-MIF group was significantly lower than that in control groups (P ＜0.05),while the expression of vimentin in SiHa-pEGFP-N1-MIF group was significantly higher than that in control groups (P ＜ 0.05).Conclusions Overexpression of MIF in cervical cancer SiHa cells can promote the EMT occurrence.
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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
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Objective To investigate the expression and significance of tumor suppressor gene PTEN and oncogenes Ki- ras in endometrioid endometrial adenocarcinoma(EEC). Methods The expression level of PTEN and p21ras were detected with the S- P immunohistochemistry method in 20 cases of normal endometrium, 20 cases of uterus atypical hyperplasia endometrium and 45 cases of EEC. The relationship between the gene expression and clinicopathological features was analyzed. Results The expression level of PTEN decreased progressively, and was 95.0 %, 40.0 %, 31.1 % respectively in normal, atypical hyperplasia endometrium and EEC. The expression level of PTEN was significantly higher than in normal in atypical hyperplasia and EEC(P
