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Objective To explore the effects of diallyl disulfide(DADS)-induced G2/M phase arrest on proliferation and apoptosis of ovarian cancer cells and its possible molecular mechanism.Methods DADS was used to incubate SK-OV-3 and OVCAR-3 cells,respectively,in different concentrations. Cell proliferation was measured by MTT assay and cell apoptosis rate was detected by flow cytometry assay. Xenograft model assay were performed to analyze the antitumor effect in vivo. Cell cycle phase distribution was detected by flow cytometry. Expressions of cell cycle G2/M phase as well as proliferation- and apoptosis-related proteins were measured by Western blotting.Results MTT assay showed that,after treatment of SK-OV-3(F=247.86,P=0.000)and OVCAR-3 cells(F=302.54,P=0.000)with different concentrations of DADS,the cell proliferation inhibition rate was significantly elevated with the increase of DADS concentrations in a concentration-dependent manner. The inhibition rate of SK-OV-3(F=335.12,P=0.000)and OVCAR-3 cells(F=347.43,P=0.000)at 24 h was significantly higher than that at 12 h and 48 h,showing a significant time-dependence manner. Flow cytometry showed that,after SK-OV-3 and OVCAR-3 cells were treated with different concentrations of DADS,the apoptosis rates increased significantly with the increase of DADS concentration in a concentration-dependent manner(P<0.05). The apoptotic rates of SK-OV-3 and OVCAR-3 cells treated with DADS at 24 h was significantly higher than that at 12 h and 48 h in a significant time-dependence manner(P<0.05). Compared with the blank treatment group,intraperitoneal injection of DADS solution significantly inhibited the xenograft volume of ovarian cancer cells in nude mice(F=548.23,P=0.000;F=311.84,P=0.000). After 30 mg/L of DADS was applied to SK-OV-3 and OVCAR-3 cells for 24 h,the percentage of cells in G2 phase of SK-OV-3 and OVCAR-3 cells increased significantly(F=375.11,P=0.000;F=256.48,P=0.000),compared with the blank cells. After 30 mg/L DADS was applied to SK-OV-3 and OVCAR-3 cells for 24 h,the expressions of p-Chk1(ser345)(F=108.89,P=0.013;F=97.58,P=0.018),p-CDC25C(ser216)(F=87.25,P=0.025;F=114.25,P=0.009),p-P53(ser15)(F=112.41,P=0.011;F=255.87,P=0.000),P21WAF1(F=246.38,P=0.001;F=141.36,P=0.005)and p-CDK1(Thr14/Tyr15)protein(F=298.12,P=0.000;F=233.15,P=0.000)were significantly increased,whereas the expressions of CDK1(F=308.24,P=0.000;F=257.55,P=0.000)and CyclinB1 protein(F=223.15,P=0.001;F=241.28,P=0.000)were significantly reduced.The expressions of proliferation and apoptosis-related proteins PCNA(F=77.36,P=0.031;F=157.28,P=0.001),Ki-67(F=205.64,P=0.007;F=315.22,P=0.000)and Survivin(F=122.13,P=0.013;F=188.24,P=0.000)were significantly decreased and Cleaved-caspase3 protein was significantly increased(F=86.46,P=0.023;F=99.11,P=0.009).Conclusion DADS can inhibit the proliferation of ovarian cancer cells and induce their apoptosis,which may be related to the activation of Chk1-CDC25C and P53-P21WAF1 signaling pathways in G2/M checkpoint,decreased kinase activity of CDK1,down-regulated expressions of CDK1 and CyclinB1 proteins,and ultimately cell cycle arrest at G2/M phase.
Subject(s)
Animals , Female , Humans , Mice , Allyl Compounds , Apoptosis , Carcinoma, Ovarian Epithelial , Cell Cycle Checkpoints , Cell Line, Tumor , Cell Proliferation , Disulfides , Mice, NudeABSTRACT
Objective To investigate the effect of vibration on the quality of suspended erythrocytes during transportation, and explore suitable experimental vibration conditions.Methods Three intensities(highway truck vibration, random vibration of fixed goods in a caterpillar, and combined wheeled vehicle vibration)were selected to simulate suspended erythrocyte transportation using electromagnetic vibration test system.Suspended erythrocytes of the same storage were randomly divided into three groups:highway truck vibration(group1),random vibration of fixed goods in a caterpillar(group 2),and combined wheeled vehicle vibration(group 3).The suspended erythrocytes were stored for 11 days and 26 days.The control group was stored with conventional methods.Suspended erythrocytes were vibrated for 1 hour,samples were collected before and after vibration,while free hemoglobin(FHb),K+,and LDH were tested.Results The changes in FHb and LDH after vibration were gradually increased with the magnitude of vibration(P<0.05).There was no significant difference in K +between the three vibration levels.The increase in FHb in suspended erythrocytes stored for 26 days was higher than 11days after random vibration of fixed goods in a caterpillar and combined wheeled vehicle vibration(P<0.05).There was no significant difference in the change in FHb between day(d)26 and d 11 after highway truck vibration.Under the same magnitude of vibration,the change in LDH in the d 26 suspended erythrocytes was more significant than that of the d 11, and no significant difference was found in the change in K +between d 26 and d 11. Conclusion The damage to suspended erythrocytes after combined wheeled vehicle vibration is more obvious than that by the other two vibrations.The vibration environment of combined wheel vehicles is more suitable for simulating the vibration damage to suspended erythrocyte vibration.Suspended erythrocytes with a longer storage time have significant changes in FHb and LDH after transportation vibration, and the length of storage time of suspended red blood cells might affect the vibration injury.
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Objective To study the effect of vibration on free hemoglobin(FHb), lactic dehydrogenase(LDH), and K+concentration of red blood cells(RBCs)stored for different lengths of time.Methods Thirty-five bags of RBCs stored for different lengths of time(3,7,11,26,and 35 d,7 bags per group)were chosen as the target of research.A wheeled vehicle that could vibrate in three directions(horizontal,longitudinal,and vertical)was used to simulate RBC transportation in addition to an electromagnetic vibration test system.The RBC samples were prepared and vibrated for 0 h,or 3 h for detection and analysis of their FHb, LDH, and K+concentration.Results The FHb concentration was significantly increased after vibration(P<0.05), and tended to increase with the extension of storage time.LDH(P<0.05)and K +concentrations(P<0.05)were obviously increased with the length of storage(3,7,11,26,and 35 d), and significantly increased after vibration of 3 h.Conclusion FHb concentration is increased after vibration, and the storage time could change FHb.The vibration and storage time have great effect on LDH and K +concentrations.
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<p><b>OBJECTIVE</b>To investigate the expression of metastasis-associated in colon cancer-1 (MACC1) in different International Federation of Gynecology and Obstetrics(FIGO)stages of epithelial ovarian cancer and its relationship with prognosis.</p><p><b>METHODS</b>Between May 2008 and August 2010, 52 epithelial ovarian cancer patients were selected from the Department of Obstetrics and Gynecology of the First Affiliated Hospital of Zhengzhou University. Reverse transcription polymerase chain reaction (RT-PCR) and Western blotting were used to detect the expression of MACC1 mRNA and protein in the primary lesions of epithelial ovarian cancer patients, the levels of MACC1 in different stage patients were compared, and the relationship between expression of MACC1 and prognosis of ovarian cancer patients was analyzed by Kaplan-Meier analysis.</p><p><b>RESULTS</b>The relative expression levels of MACC1 mRNA in epithelial ovarian cancer from 1 stage to 4 stage were 0.72±0.01, 0.75±0.01, 0.78±0.01, and 0.81±0.02, respectively (F=51.305, P=0.000). The expression levels of MACC1 protein from 1 stage to 4 stage were 0.71±0.04, 0.73±0.02, 0.76±0.01, and 0.84±0.05, respectively (F=65.142, P=0.000). At the end of the follow-up, the expression of MACC1 protein in recurrence and dead patients of 3-4 stages was obviously higher than that in the patients with stable disease (0.85±0.03 vs.0.74±0.05, F=72.324, P=0.000). Compared to 1-2 stage patients with lower MACC1 expression, the survival time of 3-4 stage patients with higher MACCC1 expression was significantly shorter (χ(2)=29.804, P=0.000).</p><p><b>CONCLUSIONS</b>Increased expression of MACC1 may indicate poor prognosis of ovarian cancer patients. Therefore, MACC1 may be a potential biomarker for advanced ovarian cancer.</p>
Subject(s)
Adult , Aged , Female , Humans , Middle Aged , Young Adult , Kaplan-Meier Estimate , Neoplasm Staging , Neoplasms, Glandular and Epithelial , Diagnosis , Metabolism , Ovarian Neoplasms , Diagnosis , Metabolism , Prognosis , RNA, Messenger , Transcription Factors , MetabolismABSTRACT
Resistance to chemotherapy is a major obstacle for the effective treatment of advanced ovarian cancer. The mechanism of chemoresistance is still poorly understood. Recently, more and more evidence showed microRNAs (miRNAs) modulated many key molecules and pathways involved in chemotherapy. microRNA-106a (miR-106a) has been implicated in many cancers, but its role in ovarian cancer and drug resistance still remains unexplored. This study was to investigate whether miR-106a mediated resistance of the ovarian cancer cell line A2780 to the chemotherapeutic agent cisplatin (DDP). The different levels of miR-106a in A2780 cells and their resistant variant A2780/DDP cells were identified by using real-time PCR. MTT assay and flow cytometry were used to analyze the effect of miR-106a on cisplatin resistance of these paired cells. Real-time PCR, Western blotting and luciferase reporter assay were applied to explore whether Mcl-1 was a target of miR-106a. As compared to A2780 cells, the expression of miR-106a was down-regulated in the cisplatin resistant cell line A2780/DDP. Moreover, knockdown of miR-106a dramatically decreased antiproliferative effects and apoptosis induced by cisplatin in A2780 cells, while overexpression of miR-106a significantly increased antiproliferative effects and apoptosis induced by cisplatin in A2780/DDP cells. Furthermore, miR-106a inhibited cell survival and cisplatin resistance through downregulating the expression of Mcl-1. Mcl-1 was a direct target of miR-106a. These results suggest that miR-106a may provide a novel mechanism for understanding cisplatin resistance in ovarian cancer by modulating Mcl-1.
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Ovarian cancer is the fifth lethal gynecologic malignancy. Metastasis-associated gene 1 (MTA1) is overexpressed in many malignant tumors with high metastatic potential. This study investigated whether down-regulation of MTA1 expression by RNAi in A2780 ovarian cancer cells could affect proliferation, anoikis, migration, invasion and adhesion of the cells and to research the potential for MTA1 gene therapy of ovarian cancer. After transfection with effective Mta1 gene siRNA, the effects on proliferation, anoikis, migration, invasion and adhesion of A2780 cells were tested by MTT assay, flow cytometry, wound-healing assay, Transwell assay and adhesion assay. Expression levels of PTEN, beta 1 integrin, MMP-9, phosphor-AKT (Ser473), and total AKT activity were evaluated in control and transfected cells. The results showed that inhibition of MTA1 mediated by Mta1-siRNA transfection decreased the cell invasion, migration and adhesion, and induced the increased cell anoikis, but no significant difference was found in proliferation of A2780 cancer cells. In addition, beta 1 integrin, MMP-9, and phosphor-AKT protein levels were significantly down-regulated, while PTEN was significantly up-regulated. These results demonstrated that MTA1 played an important role in the cell metastasis in ovarian cancer. MTA1 could serve as another novel potential therapeutic target in ovarian cancer.
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Ovarian cancer is the fifth lethal gynecologic malignancy. Metastasis-associated gene 1 (MTA1) is overexpressed in many malignant tumors with high metastatic potential. This study investigated whether down-regulation of MTA1 expression by RNAi in A2780 ovarian cancer cells could affect proliferation, anoikis, migration, invasion and adhesion of the cells and to research the potential for MTA1 gene therapy of ovarian cancer. After transfection with effective Mta1 gene siRNA, the effects on proliferation, anoikis, migration, invasion and adhesion of A2780 cells were tested by MTT assay, flow cytometry, wound-healing assay, Transwell assay and adhesion assay. Expression levels of PTEN, beta 1 integrin, MMP-9, phosphor-AKT (Ser473), and total AKT activity were evaluated in control and transfected cells. The results showed that inhibition of MTA1 mediated by Mta1-siRNA transfection decreased the cell invasion, migration and adhesion, and induced the increased cell anoikis, but no significant difference was found in proliferation of A2780 cancer cells. In addition, beta 1 integrin, MMP-9, and phosphor-AKT protein levels were significantly down-regulated, while PTEN was significantly up-regulated. These results demonstrated that MTA1 played an important role in the cell metastasis in ovarian cancer. MTA1 could serve as another novel potential therapeutic target in ovarian cancer.
Subject(s)
Female , Humans , Apoptosis , Genetics , Carcinoma , Genetics , Pathology , Cell Line, Tumor , Cell Survival , Genetics , Gene Targeting , Methods , Genetic Therapy , Methods , Histone Deacetylases , Genetics , Ovarian Neoplasms , Genetics , Pathology , Therapeutics , RNA, Small Interfering , Genetics , Therapeutic Uses , Repressor Proteins , Genetics , Treatment OutcomeABSTRACT
Resistance to chemotherapy is a major obstacle for the effective treatment of advanced ovarian cancer. The mechanism of chemoresistance is still poorly understood. Recently, more and more evidence showed microRNAs (miRNAs) modulated many key molecules and pathways involved in chemotherapy. microRNA-106a (miR-106a) has been implicated in many cancers, but its role in ovarian cancer and drug resistance still remains unexplored. This study was to investigate whether miR-106a mediated resistance of the ovarian cancer cell line A2780 to the chemotherapeutic agent cisplatin (DDP). The different levels of miR-106a in A2780 cells and their resistant variant A2780/DDP cells were identified by using real-time PCR. MTT assay and flow cytometry were used to analyze the effect of miR-106a on cisplatin resistance of these paired cells. Real-time PCR, Western blotting and luciferase reporter assay were applied to explore whether Mcl-1 was a target of miR-106a. As compared to A2780 cells, the expression of miR-106a was down-regulated in the cisplatin resistant cell line A2780/DDP. Moreover, knockdown of miR-106a dramatically decreased antiproliferative effects and apoptosis induced by cisplatin in A2780 cells, while overexpression of miR-106a significantly increased antiproliferative effects and apoptosis induced by cisplatin in A2780/DDP cells. Furthermore, miR-106a inhibited cell survival and cisplatin resistance through downregulating the expression of Mcl-1. Mcl-1 was a direct target of miR-106a. These results suggest that miR-106a may provide a novel mechanism for understanding cisplatin resistance in ovarian cancer by modulating Mcl-1.
Subject(s)
Female , Humans , Antineoplastic Agents , Pharmacology , Cell Line, Tumor , Cisplatin , Pharmacology , Drug Resistance, Neoplasm , Genetics , MicroRNAs , Genetics , Myeloid Cell Leukemia Sequence 1 Protein , Genetics , Ovarian Neoplasms , Drug Therapy , GeneticsABSTRACT
<p><b>OBJECTIVE</b>To analyze GJB6 gene mutations in a Chinese family with hidrotic ectodermal dysplasia and to provide first-trimester prenatal diagnosis for a fetus.</p><p><b>METHODS</b>Mutation scanning was carried out with PCR and bilateral direct sequencing in 2 affected and 6 unaffected individuals from the family. After the mutation was confirmed, prenatal diagnosis was performed on chorionic villi samples obtained at 11th gestational week.</p><p><b>RESULTS</b>A heterozygous missense mutation c.31G>A of the GJB6 gene was discovered in all of the patients, which has led to substitution of glycine by arginine at codon 11 (p.G11R) at the N-terminal of the GJB6 protein. Prenatal diagnosis indicated that the fetus had also carried the same p.G11R mutation. Following termination of the pregnancy, analysis of the aborted tissues was consistent with prenatal diagnosis.</p><p><b>CONCLUSION</b>The missense mutation c.31G>A(p.G11R) of the GJB6 gene probably underlies the disease in this family. Prenatal diagnosis with DNA sequencing can facilitate genetic counseling of this family.</p>
Subject(s)
Adult , Child , Child, Preschool , Female , Humans , Male , Pregnancy , Asian People , Genetics , Base Sequence , DNA Mutational Analysis , Ectodermal Dysplasia , Diagnosis , Embryology , Genetics , Fetal Diseases , Diagnosis , Genetics , Molecular Sequence Data , Pedigree , Pregnancy Trimester, First , Genetics , Prenatal DiagnosisABSTRACT
<p><b>OBJECTIVE</b>To analyze the mutation of IL2RG gene in a Chinese family with a birth history of a dead child suspected of X-linked severe combined immunodeficiency (X-SCID), and to perform prenatal diagnosis with DNA sequencing.</p><p><b>METHOD</b>Blood samples of the parents of the dead child and chorionic villi at gestational age 11 weeks were collected. Eight exons comprising the open reading frame as well as their exon/intron boundaries of IL2RG gene were analyzed by PCR and bi-directional sequencing.</p><p><b>RESULT</b>A heterozygous nucleotide substitution c.690C > T (R226C) in exon 5 was detected in the mother, but not in the father. In the second pregnancy of the mother, the mutation of R226C was not detected in the male fetus by prenatal diagnosis, and the heterozygous mutation was detected in the female fetus of the third pregnancy. The reliability of the prenatal genetic diagnosis was confirmed by the one-year follow-up after the neonates were born.</p><p><b>CONCLUSION</b>The mutation of c.690C>T in IL2RG gene may be the pathologic cause of the proband with X-SCID. DNA sequencing combining sex determination is a valid strategy for prenatal diagnosis of X-SCID.</p>
Subject(s)
Adult , Female , Humans , Infant , Male , Pregnancy , Asian People , Genetics , Base Sequence , DNA Mutational Analysis , DNA Primers , Exons , Genetics , Heterozygote , Interleukin Receptor Common gamma Subunit , Genetics , Mutation , Pedigree , Polymerase Chain Reaction , Prenatal Diagnosis , Methods , X-Linked Combined Immunodeficiency Diseases , Diagnosis , GeneticsABSTRACT
<p><b>BACKGROUND</b>Invasive cancer of the cervix is considered a preventable disease because it has a long pre-invasive state, cervical cytology screening programs are currently available, and treatment of pre-invasive lesions is effective. We tested the accuracy of frozen section examination (FSE) of cone specimens to identify the endocervical margin and rule out invasion in patients with high-grade cervical intraepithelial neoplasia (CIN).</p><p><b>METHODS</b>For 320 consecutive patients with a preoperative biopsy result of CIN stage 2/3, cold-knife conization (CKC) was performed followed by FSE. The results from analyses of permanent paraffin sections (PS) were compared with the FSE findings.</p><p><b>RESULTS</b>The accuracy of FSE was 87% (278/320). For all of the seven patients with an invasive squamous cell carcinoma of the cervix identified by FSE, the diagnosis was confirmed by PS analysis. For one patient, the FSE result was cervicitis, whereas PS analysis showed microinvasive carcinoma. Appropriate surgery was performed for all patients based on the FSE and biopsy results. The FSE and PS results were not significantly different (P = 0.000). Definitive examination of margin status using PS was concordant with FSE findings in all cases.</p><p><b>CONCLUSIONS</b>FSE is a rapid and reliable method for evaluating CKC specimens. It can identify frank invasion, permit adequate treatment in a one-stage procedure, and reliably detect clear resection margins. Since discrepancies do exist and may result in inappropriate treatment, further research is required to decrease these discrepancies and avoid missing even one case.</p>
Subject(s)
Adult , Aged , Female , Humans , Middle Aged , Young Adult , Uterine Cervical Dysplasia , Diagnosis , Frozen Sections , Methods , In Vitro Techniques , Prospective StudiesABSTRACT
<p><b>OBJECTIVE</b>To evaluate the feasibility of genetic analysis of tyrosinase gene (TYR) in oculocutaneous albinism type I (OCA1). Mutation analysis and prenatal genetic diagnosis of TYR gene for seven pedigrees with OCA1 were performed.</p><p><b>METHODS</b>PCR was used to amplify the exons, exon-intron boundaries and promoter of the TYR gene in the probands and/or their parents. The products were further analyzed by direct sequencing. Prenatal genetic diagnoses were performed by chorionic villus sampling after the genotypes of the probands or their parents were determined.</p><p><b>RESULTS</b>Compound heterozygous mutations were detected in all pedigrees, which included 9 mutations, namely R76Q, c.232insGGG, R116X, R278X, R299H, c.929-930insC, IVS2-11delTT, Q399X and W400L. Among these, R76Q and Q399X were identified for the first time. Seven families have requested prenatal diagnoses. One fetus was detected with double mutations of TYR gene, and the parents have decided to have therapeutic abortion. Two fetuses did not carry the mutations identified in the probands, whilst other four fetuses were carriers of heterozygous mutations. Six families decided to carry on with the pregnancies. And the neonates did not show any symptoms of OCA after birth.</p><p><b>CONCLUSION</b>Direct sequencing of the TYR gene is helpful for genetic counseling, prenatal diagnosis and carriers screening of OCA1.</p>
Subject(s)
Female , Humans , Infant, Newborn , Male , Pregnancy , Albinism, Oculocutaneous , Diagnosis , Genetics , Genetic Predisposition to Disease , Monophenol Monooxygenase , Genetics , Mutation , Pedigree , Prenatal Diagnosis , MethodsABSTRACT
<p><b>OBJECTIVE</b>To identify potential mutations of ED1 gene in six pedigrees with hypohidrotic ectodermal dysplasia (HED), and to provide genetic counseling and prenatal diagnosis.</p><p><b>METHODS</b>Eight coding exons of ED1 gene of patients with clinically diagnosed HED and their relatives were amplified by polymerase chain reaction (PCR). The products were further analyzed by direct sequencing.</p><p><b>RESULTS</b>Various mutations of ED1 gene were detected, which included R153C, A349T, G299S, A349T and X392Q. Heterozygous double peaks at the same position were found in female carriers. Deletion of exon 9 was detected in one pedigree. R153C, X392Q and deletion of exon 9 were first identified in ethnic Han Chinese.</p><p><b>CONCLUSION</b>The identified mutations of ED1 gene may be responsible for the disease. Genetic counseling, prenatal diagnosis and carrier screening are now available for these families.</p>
Subject(s)
Adolescent , Adult , Child , Child, Preschool , Female , Humans , Male , Base Sequence , China , Ectodermal Dysplasia , Genetics , Ectodysplasins , Genetics , Genetic Predisposition to Disease , Heterozygote , Molecular Sequence Data , Mutation , PedigreeABSTRACT
<p><b>OBJECTIVE</b>To investigate the expressions of metastasis-associated in colon cancer-1 (MACC1), hepatocyte growth factor (HGF), and C-met proteins in epithelial ovarian cancer and their significance.</p><p><b>METHODS</b>The expressions of MACC1, HGF and C-met in 20 specimens of normal ovarian tissues, 19 specimens of benign epithelial ovarian tumor and 52 specimens of epithelial ovarian cancer were measured by immunohistochemistry and Western blotting. The correlations of the expressions of MACC1, HGF and C-met protein to the clinicopathologic characteristics of epithelial ovarian cancer were analyzed, and the correlations between the expressions of the 3 proteins were also evaluated.</p><p><b>RESULTS</b>The positivity rates of MACC1, HGF and C-met proteins were 73.1%, 63.5% and 78.8% in epithelial ovarian cancer with relative expressions of 0.72∓0.05, 0.64∓0.04 and 0.79∓0.04, respectively, showing significant differences from those in normal ovarian tissues and benign ovarian tumors (P<0.05). In epithelial ovarian cancer, the up-regulation of MACC1, HGF and C-met expressions were associated with advanced FIGO stage, poor differentiation and lymph node metastasis (P<0.05). MACC1 expression was positively correlated to HGF (r=0.350, P=0.011) and C-met expressions (r=0.429, P=0.002), and the latter two was also positively correlated (r=0.487, P=0.000).</p><p><b>CONCLUSIONS</b>MACC1 may serve as a potential biomarker for advanced ovarian cancer. Deregulation of MACC1, HGF and C-met proteins may synergistically participate in the malignant progression of epithelial ovarian cancer.</p>
Subject(s)
Adult , Aged , Female , Humans , Middle Aged , Biomarkers, Tumor , Metabolism , Hepatocyte Growth Factor , Metabolism , Neoplasms, Glandular and Epithelial , Metabolism , Pathology , Ovarian Neoplasms , Metabolism , Pathology , Ovary , Metabolism , Pathology , Proto-Oncogene Proteins c-met , Metabolism , Transcription Factors , MetabolismABSTRACT
<p><b>OBJECTIVE</b>To study the characteristics of the phenylalanine hydroxylase gene (PAH) mutations in patients with phenylketonuria (PKU) in Henan province, in order to provide basic information for genetic counseling and prenatal diagnosis.</p><p><b>METHODS</b>Mutations of the PAH gene were detected in exons 1-13 with flanking introns of PAH gene by PCR and DNA sequencing in 47 families with PKU.</p><p><b>RESULTS</b>A total of 25 different mutations were detected in 83 out of 94 PAH alleles (88.3%). Among them, E79fX13, H271R and D415Y have not been reported previously. It was the first time that IVS10-14C to G mutation was reported in Chinese PKU population. The mutations p.R243Q, EX6-96A to G, p.Y356X, IVS401G to A, p.R111X, p.V399V and p.R413P, were the prevalent mutations with relative frequencies of 20.5%, 12.0%, 9.6%, 9.6%, 8.4%, 8.4% and 7.2% respectively.</p><p><b>CONCLUSION</b>The mutations of the PAH gene in patients with classical phenylketonuria in Henan province were similar to that in other areas of China. Prenatal gene diagnosis for PKU by PAH gene sequencing is efficient for most PKU families.</p>
Subject(s)
Child, Preschool , Female , Humans , Male , Base Sequence , China , DNA Mutational Analysis , Methods , Genetic Counseling , Methods , Molecular Sequence Data , Phenylalanine Hydroxylase , Genetics , Phenylketonurias , Diagnosis , Genetics , Polymerase Chain Reaction , Methods , Prenatal Diagnosis , Methods , Sequence Analysis, DNA , MethodsABSTRACT
<p><b>OBJECTIVE</b>To detect the expression of human similar expression to FGF gene(hSef) and fibroblast growth factor-2(FGF-2) and their correlation with epithelial ovarian tumor.</p><p><b>METHODS</b>Immunohistochemical SP staining was used to detect the expression of hSef and FGF-2 proteins in 31 cases of epithelial ovarian carcinoma (EOC), 18 cases of benign epithelial tumor (BET), 10 cases of normal ovarian (NO) tissues collected from July 2007 to May 2008. The expression of hSef mRNA in 24 cases of EOC, BET and NO collected from July 2008 to May 2009 were analyzed by RT-PCR.</p><p><b>RESULTS</b>The results of immunohistochemical study showed that the expression of hSef in the EOC tissues were significantly lower than that in the NO and BET (P < 0.001). However, the expression of FGF-2 was higher (P = 0.002). The expression of hSef had a negative correlation with FGF-2 (r(s) = -0.324, P = 0.012). The RT-PCR results showed that there was a gradually declined trend of expression of hSef in NO, BET to EOC (P < 0.001), but the expression of FGF-2 in NO, BET to EOC was gradually increased (P < 0.001), with a significant negative correlation (NO: r(s) = -0.910, P < 0.001; BET: r(s) = -0.859, P < 0.001; EOC: r(s) = -0.888, P < 0.001).</p><p><b>CONCLUSIONS</b>The expression of hSef is decreased in epithelial ovarian carcinoma tissue, but the expression of FGF-2 is increased. It is likely that low hSef expression is related to the the carcinogenesis and development of epithelial ovarian carcinoma by suppressing the promoting effects of FGF-2 to cell proliferation.</p>
Subject(s)
Adult , Aged , Female , Humans , Middle Aged , Cystadenocarcinoma, Mucinous , Genetics , Metabolism , Pathology , General Surgery , Cystadenocarcinoma, Serous , Genetics , Metabolism , Pathology , General Surgery , Cystadenoma, Mucinous , Genetics , Metabolism , Pathology , General Surgery , Cystadenoma, Serous , Genetics , Metabolism , Pathology , General Surgery , Fibroblast Growth Factor 2 , Genetics , Metabolism , Gene Expression Regulation, Neoplastic , Immunohistochemistry , Ovarian Neoplasms , Genetics , Metabolism , Pathology , General Surgery , Ovary , Metabolism , RNA, Messenger , Metabolism , Real-Time Polymerase Chain Reaction , Receptors, Interleukin , Genetics , MetabolismABSTRACT
<p><b>BACKGROUND</b>Bacterial vaginosis (BV) is one of the most common infectious diseases among sexually active women and is associated with the increased acquisition of a variety of sexually transmitted diseases. This study aimed to compare the efficacy of a non-antibiotic sucrose gel against an antibiotic metronidazole gel for the treatment of BV.</p><p><b>METHODS</b>A randomized, double-blinded, multi-center, parallel-group, placebo-controlled phase III clinical trial was conducted at eight hospitals in China. A total of 560 subjects with clinically diagnosed BV were randomly assigned into three groups for vaginally receiving sucrose, metronidazole, and placebo gels, respectively, twice daily for five consecutive days. The efficacy of therapeutic cure, defined as an achievement of both microbiologic cure (a Nugent score of 3 or less) and clinical cure (a resolution of the clinical findings from the baseline visit), was evaluated at the 1st and 2nd test-of-cure (TOC) visits at 7-10 and 21-35 days after the start of treatment, respectively.</p><p><b>RESULTS</b>Therapeutic cure rates for sucrose, metronidazole, and placebo gel groups were 83.13%, 71.30% and 0.92%, at the 1st TOC, and 61.04%, 66.67% and 7.34%, at the 2nd TOC, respectively. While there was no significant difference between the sucrose and metronidazole gel groups at the 2nd TOC (P = 0.305), and sucrose gel was more effective than metronidazole gel at the 1st TOC (P = 0.009).</p><p><b>CONCLUSION</b>These findings suggest that sucrose gel restores normal vaginal flora more rapidly than metronidazole gel and can be used as a novel treatment for BV.</p>
Subject(s)
Adolescent , Adult , Female , Humans , Middle Aged , Young Adult , Administration, Intravaginal , Anti-Bacterial Agents , Therapeutic Uses , Double-Blind Method , Metronidazole , Therapeutic Uses , Sucrose , Therapeutic Uses , Treatment Outcome , Vaginosis, Bacterial , Drug TherapyABSTRACT
<p><b>OBJECTIVE</b>To detect the expression of the protein of TGF-beta1 and E-cadherin in the primary and metastatic lesions of ovarian carcinoma and explore the mechanism of the metastasis of ovarian carcinoma.</p><p><b>METHODS</b>Immunohistochemistry (IHC) was performed to detect the expression of TGF-beta1 and E-cadherin proteins in primary and metastatic ovarian carcinoma, benign epithelial ovarian tumor and normal ovarian tissue.</p><p><b>RESULTS</b>The expression of TGF-beta1 was significantly higher in ovarian carcinoma (67.2%) than in benign tumors (28.6%) and normal ovarian tissue (18.9%) (Chi2=26.94, P<0.001), but E-cadherin expression showed a reverse pattern. TGF-beta1 expression in the primary ovarian carcinoma carcinoma was associated with the FIGO stage, lymph metastasis and ascites of the tumor (P=0.01, P=0.01, and P=0.04, respectively). E-cadherin expression in the tumor was associated with the differentiation (P=0.02) and lymph metastasis of ovarian carcinoma (P=0.04). The expressions of TGF-beta1 and E-cadherin were all significantly lower in the primary tumors than in the metastatic tumor (Chi2=4.70, P=0.03; Chi2=5.91, P=0.015). A significant correlation was found between the expressions of the TGF-beta1 and E-cadherin in the primary carcinoma (Kappa value of -0.32, P=0.01).</p><p><b>CONCLUSION</b>TGF-beta1 and E-cadherin are closely associated with the metastasis of ovarian carcinoma and might be potential targets for controlling the metastasis of ovarian carcinoma.</p>
Subject(s)
Adult , Female , Humans , Middle Aged , Cadherins , Genetics , Metabolism , Lymphatic Metastasis , Neoplasm Metastasis , Ovarian Neoplasms , Metabolism , Pathology , Peritoneal Neoplasms , Metabolism , Transforming Growth Factor beta1 , Genetics , MetabolismABSTRACT
Objective To investigate the characteristics of the preoperative and postoperative urodynamical parameters of women with uterine cervical carcinoma after radical hysterectomies.Methods Forty-six women had uterine cervical carcinoma at stage Ⅰ b or Ⅱ a.Complete pre-and postoperative urodynamie follow-ups were conducted for each patient.Results Twenty-six women(57%)who had preoperatively normal urinary tract function needed to void by abdominal straining after radical surgery.After the radical hysterectomy,the postvoid residual volume[(205?201)vs(5?3)ml,P