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Objective:To identify the expression level of known microRNA (miRNA) by high-throughput sequencing and analysis of the miRNA of Schistosoma japonicum schistosomula, and predict the miRNA target genes and their biological functions. Methods:Schistosoma japonicum schistosomula were prepared in vitro, and total RNA of schistosomula were extracted and analyzed to construct a library for performing high-throughput sequencing. The difference of miRNA expression was analyzed by using DEGseq R language package and perl script. Then the target genes and their biological functions of differential miRNA were predicted by miRanda software, Blast software, and KEGG database respectively. Results:There were 38 483 matching sequences in the miRNA expressed of Schistosoma japonicum schistomula in the constructed library compared with the latest miRBase database, and 60 miRNA were identified, sja-miR-125b was the miRNA with the highest expression, followed by sja-miR-61, sja-miR-71a, sja-miR-36-3p and sja-miR-10-5p, which accounted for 91% (3 263/3 585) of the total miRNA expression. MiRanda software predicted a total of 7 176 target genes, gene functions concentrated on nucleotide transferase activity, cellular nitrogen complex metabolism, molecular function, biological processes, biosynthesis, plasma membrane and protein maturation. The functional enrichment analysis showed that the highly expressed miRNA were mainly involved in pathogenic process, biological progress and multiple metabolic regulation pathways. Conclusions:The miRNA expressed significantly of Schistosoma japonicum schistomula mainly involved in the regulation of metabolic pathways during the differentiation, growth and pathogenesis of Schistosoma japonicum. To lay a foundation for the study of the regulatory mechanism of Schistosoma japonicum development and the development of new drugs.
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Objective@#To compare the carcinogenic abilities of CD133+CD44+ laryngeal cancer stem cells and general laryngeal cancer stem cells and to identify the mechanism underlying the action of miRNAs.@*Methods@#Solid tumor-derived laryngeal carcinoma stem cells and Hep-2-derived laryngeal carcinoma stem cells were cultured, and CD133+CD44+ laryngeal cancer stem cells were sorted by flow cytometry. Boden chamber invasion assay, cell migration assay and tumor formation assay were then performed to compare the invasion, migration and tumorigenic abilities of CD133+CD44+ laryngeal cancer stem cells and general laryngeal cancer stem cells. And then, miRNAs isolated from two laryngeal cancer stem cells were detected and analysed with miRNA chip.@*Results@#(1)In Boyden chamber invasion assay, the cell invasion rate of CD133+CD44+ laryngeal cancer stem cells was obviously higher (80.2%±2.3% vs. 63.9%±3.2%, t=5.011, P=0.027); (2)CD133+CD44+ laryngeal cancer stem cells also had higher mobility in cell migration assay (82.9%±1.1% vs. 70.9%±0.6%, t=4.514, P=0.031); (3)In tumor formation assay, the tumor formation rate of CD133+CD44+ laryngeal cancer stem cells was also higher (80% vs. 50%). What′s more, we identified 15 miRNAs that were significantly upregulated in CD133+CD44+ laryngeal cancer stem cells and 3 miRNAs that were significantly downregulated in CD133+CD44+ laryngeal cancer stem cells, compared with normal laryngeal cancer stem cells.@*Conclusions@#CD133+CD44+ laryngeal cancer stem cells have stronger invasion, migration and tumorigenic abilities compared with normal laryngeal cancer stem cells, and the difference of miRNAs′ expression is one of the possible causes.
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Objective To understand the spatial distribution characteristics of wild feces in schistosomiasis endemic areas of Jiangling County,Hubei Province and further explore the source of infection efficiently,so as to provide the evidence for the development of corresponding monitoring and response technology. Methods In 2011,the fresh wild feces were investigated every two months in the selected 15 villages by the severity of historical endemic in Jiangling County. The schistosome miracidi-um hatching method was used to test the schistosome infection of the wild feces. The descriptive analysis and spatial analysis were used for the description of the spatial distribution of the wild feces. Results Totally 701 wild feces samples were collected with the average density of 0.0556/100 m2,and the positive rate of the wild feces was 11.70%(82/701). The results of the re-gression analysis showed a positive spatial correlation between the positive rate of wild feces and the rate of human infection,the area with infected Oncomelania hupensis and the number of fenced cattle,and the corrected R2 of the model was 0.58. Conclu-sion The infection rate of wild feces is positively correlated with the rate of human infection,area with infected O. hupensis and number of fenced cattle in space in Jiangling County,so the prevention and control measures could be conducted according to the spatial distribution of the positive wild feces.
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Objective To detect the value of circulating tumor cells(CTCs)in peripheral venous blood in patients with metastatic breast cancer. Methods 50 female patients with advanced advanced metastatic breast can-cer hospitalized in our hospital from May 2016 to December 2016 were enrolled in the research. Patients were divid-ed into oligometastases group and extensive metastasis group through multi-department comprehensive analysis andimaging diagnosis. 10 cases with early stage breast cancer were selected randomly. 10 healthy female volunteers were recruited as control group. After obtaining written informed consent from research subjects ,7.5 mL peripheral blood was drew from patients and volunteers prior to starting a new line of chemotherapy ,surgery or other treat-ment. CTCs counts from Blood samples were detected density gradient centrifugation associate with flow cytometry. Results The base line was formulated as CTCs≥5/7.5 mL positive and CTCs<5/7.5 mL negative. By comparing the positive expression of CTCs in early and advanced metastatic breast cancer(Pa = 0.01,P < 0.05),positive CTCs was associated with advanced metastatic breast cancer. Comparision of the positive expression of CTCs between oligometastases group and the extensive transfer group showed significant difference in the CTCs count be-tween the two groups(Pb = 0.018,P < 0.05). In the corresponding period,no positive CTCs was detected in all healthy volunteers. Conclusion CTCs count was associated with metastatic breast cancer. There was a correlation between tumor metastasis and CTCs count (the more widely metastasis ,the higher the detection rate of CTCs). CTCs may be of relevant value in the diagnosis and treatment ,and prognosis evaluation of metastatic breast cancer.
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Objective To implement a two?level community?based health education pattern of schistosomiasis in residents of endemic areas in marshland and lake regions,so as to explore the suitable pattern of health education under hypo?endemic situa?tion. Methods Two schistosomiasis endemic villages in Jiangling County,Hubei Province were collected as study areas,and among which,one village was treated as an intervention group,where the two?level community?based health education pattern as well as regular control measures was implemented;the other village was a control group,where only regular control measures were implemented. The awareness rates on schistosomiasis control,the rates of correct behavior and the compliance rates of ex?amination,treatment and chemotherapy of the two groups before and after the intervention were compared. Results According to the results of the baseline survey in 2014,the awareness rates of schistosomiasis control of the intervention and control groups were 84.00%and 77.45%,respectively,the correct rates of behavior of the two groups were 72.00%and 63.73%,respectively, and the compliance rates of the treatment were 80.36%and 82.28%,respectively,there were no statistically significant differ? ences between all the above rates of the two groups(all P>0.05). After the intervention of the two?level community?based health education,the correct rates of behavior,and the compliance rates of examination and chemotherapy of the two groups were 92.31%and 80.37%,95.11%and 82.55%,84.13%and 63.64%,respectively,and the differences between all the rates above of the two groups were statistically significant(all P<0.05). When compared to those before intervention,the growing rates of the compliance rates of examination,treatment and chemotherapy of the intervention group were 20.97%,15.33%and 23.29%, respectively,while those of control group were 14.27%,4.17%,-3.77%,respectively,the growing rates of the intervention groups were higher than those of the control groups. Conclusions Through the two?level community?based pattern of health edu?cation,the compliance rates of examination and treatment of the residents have improved,and therefore,the pattern is suitable for popularization and application in marshland and lake regions.
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Objective To observe the Oncomelania hupensis snail control effects of molluscicides with different formula?tions in the ditch with unstable water level in field of marshland and lake areas,so as to provide the evidence for formulating the prevention strategies. Methods A drainage channel with O. hupensis snails in Jiangling County,Jingzhou City were selected as the study field,then it was divided into 6 segments,except 1 segment was treated as the blank control group for observing the natural mortality rate of the snails,the other 5 segments were treated as observation groups,where 4% niclosamide ethanol?amine salt powder,5%niclosamide ethanolamine salt granule,25%suspension concentrate of niclosamide ethanolamine salt, 26% suspension concentrate of metaldehyde and niclosamide ethanolamine salt,50% niclosamide ethanolamine salt wettable powder were applied respectively. Before and after the mollusciciding,the snail surveys were carried out through the systematic sampling method,and the short?and long?term effects of snail control were observed,and the mortality rates of the snails on the slope above the water level and those below the water level were compared. Results When 7 d and 15 d after mullusciciding, the mortality rates of snails on the slope above the water level in each observation group were 79.52%-97.87% and 90.43%-96.30%,respectively,when compared with those before mollusciciding,all the differences were statistically significant(all P<0.01). When 7 d after mullusciciding,the mortality rates of snails below the water level in each observation group were 17.11%-50.00%,which were all lower than those of the snails on the slope above the water level in the corresponding groups(all P<0.05);when 15 d after mullusciciding,the rates were 9.43%-95.24%,and those in 25%suspension concentrate of niclosamide ethanolamine salt,26%suspension concentrate of metaldehyde and niclosamide ethanolamine salt,50%niclosamide ethanol?amine salt wettable powder groups were all lower than those of the snails on the slope above the water level in the corresponding groups(all P<0.01). Compared with the densities of living snails before mollusciciding,those 6 months after mollusciciding in each observation group decreased by 70.21%-78.98%. Conclusions The molluscicides in the formulation of powders and granule are suitable for both the snail environment with or without water,while those in the formulation of suspension agents and wettable powders are suitable only for environments with water.
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Objective To classify and grade the environments with Oncomelania hupensis snails inside embankment in marsh-land and lake areas,so as to improve the work efficiency and realize the systematic management. Methods The schistosomiasis endemic area Liaodi and Xiongdi villages in Jiangling County,Hubei Province were selected as the experiment villages,and the environments with snails in the two villages were divided into sections with a length of 500m,then the snail situation were sur-veyed by the systematic sampling method with 10m and 50m a frame respectively. The environments were classified according to the discovery of infected snails and wild fecal contamination,and the numbers of sites with snails and their areas in different class-es were recorded. Meanwhile,the data of sites with infected snails in Jingzhou City during the recent 5 years were collected and graded according to the average density of snails,so as to discuss the correlativity between the grades of environments with snails and the numbers of sites with infected snails and the occurrence rate of frames with snails. Results There were 1 967 sites with in-fected snails in Jingzhou City during the recent 5 years,and there was a positive correlation between the grades of environments with snails and the occurrence rate of frames with snails(rs=0.77,P0.05),but the time-consuming of the former was 3 times of the latter. Conclusions The classification and grading of the environments with snails inside embankment in marshland and lake areas can master the key point of the snail survey. The subsection and setting frames at a suitable distance can save time and manpower,improve work efficiency,as well as understand the distribution of snail status of the environment with snails,which can realize the sort management of the snail envi-ronments inside embankment.
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Objective To build a high quality diagnosis system for schistosomiasis surveillance in the situation of low infec-tion in Jianglin County. Methods The network laboratory for schistosomiasis diagnosis was built according to the national crite-ria in Jianglin County in 2012. Results The network laboratory for schistosomiasis diagnosis was established successfully and the operation was quiet well. Conclusion The establishment and operation of the laboratory play an important role in the real-ization of schistosomiasis elimination.
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Objective To evaluate the efficacy of routinely used pattern for schistosomiasis diagnosis in lake and marshland regions. Methods A historically heavy endemic village of schistosomiasis named Jinggan Village from Jiangling County was se?lected for field survey. The residents aged 6?65 years were screened by indirect hemagglutination assay(IHA)and dipstick dye immunoassay(DDIA)in parallel. The serological positives were examined by Kato?Katz technique and miracidium hatching technique to determine the infection of schistosome. The consistency of the two serological methods was evaluated. In addition , the schistosome infection rates were estimated according to the 3 detection patterns namely IHA,DDIA,IHA+DDIA combined with the etiologic examination. Results A total of 530 individuals were examined by the serological tests. The positive rate of DDIA was 46.98%(249/530),significantly higher than that of IHA(28.49%,151/530)(χ2=59.55,P<0.01). Totally 279 in?dividuals were serological positives determined by IHA or DDIA,while 252 of them were detected by stool examination,and 22 cases were determined as parasitological positives,while 7 and 3 cases were diagnosed as antibody negatives by IHA and DDIA,respectively. The estimated infection rates determined by IHA,DDIA,IHA plus DDIA combined with stool examination were 3.14%,3.97%、4.60%,respectively. Conclusions Under the condition of endemic situation becoming more and more waning,the current routinely used pattern for schistosomiasis detection may lead to missed diagnosis. So,more sensitive and ef?fective diagnostic tools or appropriate detection patterns need to be explored.
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Objective To evaluate the effect of the Parasep? feces centrifuge tube method on detecting schistosome eggs. Methods A total of 803 residents aged from 6-65 years were selected in 2 schistosomiasis endemic villages Jiangling Coun-ty Hubei Province and their stool samples were collected and detected parallelly by the Kato-Katz technique nylon silk egg hatching method and Parasep? feces centrifuge tube method at the same time. Results Among the 803 people 15 cases were found of schistosome egg positive and the positive rate was 1.87%. The positive rates of the Kato-Katz technique nylon silk egg hatching method and Parasep? feces centrifuge tube method were 0.75% 1.49% and 1.12% respectively. The schistosome eggs got with the Parasep? feces centrifuge tube method were clear and easy to identify. Conclusion In low endemic areas of schistosomiasis the Parasep? feces centrifuge tube method can be used as schistosomiasis japonica etiology diagnosis method.
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OBJECTIVE@#To investigate the expression and clinical significance of signal transducer and activator of transcription 3(STAT3) and its target gene c-myc in laryngeal squamous cell carcinoma (LSCC), and the relationship between the two genes.@*METHOD@#Formalin-fixed and paraffin-embedded tissues, which came from 56 cases of LSCC and 30 samples of normal mucosas from 30 patients with total or subtotal laryngectomy over 2.0 cm away from tumor margin, were detected for the expression of STATS, c-myc by in situ hybridization and SP immunohistochemistry. Micro-image analysis system was used to determine the optical density, and the result was analyzed statistically.@*RESULT@#There is overexpression of STAT3, c-myc mRNA and protein in LSCC. The expression of STAT3 and c-myc mRNA in LSCC was associated with clinical stage, differentiation grade and lymph nodal metastases (P < 0.05 or 0.01). The expression of STAT3 and c-myc protein in LSCC was associated with clinical stage and lymph nodal metastases (P < 0.05 or 0.01). There was a positive correlation between the expression of STAT3 and c-myc genes. mRNA and protein, The correlation coefficient (r) was 0.6224 (P < 0.01) for the mRNA expression and 0.7012 (P < 0.01 )for the protein expression.@*CONCLUSION@#The expression of STAT3 and c-myc may play an important role in the tumorigenesis, metastases and poor prognosis of LSCC. There was a positive correlation between the overexpression of STAT3 and c-myc genes in LSCC.
Subject(s)
Adult , Aged , Aged, 80 and over , Female , Humans , Male , Middle Aged , Carcinoma, Squamous Cell , Metabolism , Pathology , Laryngeal Neoplasms , Metabolism , Pathology , Lymphatic Metastasis , Neoplasm Staging , Proto-Oncogene Proteins c-myc , Metabolism , STAT3 Transcription Factor , MetabolismABSTRACT
Objective To explore the dynamic changes of diffusion tensor imaging(DTI) in spinal cord of goats with persistent compression injury. Methods Eighteen goats weighted 20--25 kg were divided into three groups with completely random design: A, B and C. A balloon catheter was inserted into the epidural space at C3-4 level via intervertabral foramen for each goat. The balloon was inflated by injection of variable volumes of saline in group A and B 10 days following operation. The volume of saline was 0. 3 ml in group A and 0. 2 ml in group B,respectively. The compression sustained for 40 days. Group C served as uncompressed control without injection of saline. The locomotor rating score was applied to each group. Conventional MRI and DTI were performed. The apparent diffusion coefficient (ADC)and fractional anisotropy (FA) values were measured. Histopathological assessments of the compressed spinal cord were performed 50 days following operation with light microscope and transmission electron microscopy. Results Before operation, the locomotor rating score was 5, the ADC value was ( 1.23 ± 0. 05 ) × 10-3 mm2/s and the FA value was (0. 72 ± 0. 05 ) each group. Of six goats in Group A, the locomotor rating score severely decreased and reached( 1.5±0. 4)on the 40 th day after compression. The ADC value at compression site decreased soon and reached the minimum (0. 75±0. 04) × 10-3mm2/s on the 5 th day after compression. Then the ADC value increased gradually, restored normal on the 10 th day or so, then became markedly higher than normal and reached( 1.61±0. 05) × 10-3mm2/s on the 40 th day. The FA value at compression site decreased soon, reached(0. 54±0. 04)on the 1st day, then decreased gradually and reached(0. 43 ± 0.05) on the 40 th day. It appeared high signal intensity on T2WI on the 10 th day. In Group B, the locomotor rating score was moderately decreased and reached(3.4±0. 5 )on the 40 th day. The ADC value at compression site decreased slightly firstly, reached( 1.08±0. 04) × 10-3mm2/s on the 1st day, restored normal on the 20 th day or so, then increased gradually, became higher than normal and reached ( 1.27 ± 0. 05) × 10-3mm2/s on the 40 th day. The FA value increased slightly firstly, reached (0. 78±0. 05 )on the 1st day, then decreased gradually, restored normal on the 15 th day or so, became lower than normal and reached(0. 67±0. 05) on the 40 th day. There was no abnormality on conventional MRI. In Group C, the locomotor rating score, ADC value and FA value remained unchanged. There was no abnormality on conventional MRI. There were dynamic changes over time of the ADC value and FA value in Group A and B, which was more marked in Group A than that in Group B ( repeated measurements analysis of variance, F=426. 7 for the ADC value and F =7895.2 for the FA value, P < 0. 01 ). Histopathologically, swelling and degeneration of axons and neurons as well as the disarrangement of myelin sheathes could be seen. The pathological changes were more marked in Group A than in Group B. In Group C, no abnormality could be seen. Conclusion There are dynamic changes of DTI in cervical spinal cord with compressive injury that correlated with the degree and duration of compression. The ADC value decreased firstly, restored normal and then increased. The FA value increased firstly, restored normal and then decreased in mild compression while solely decreased in serious compression.