1.Effects of long non-coding RNA HOTAIR on proliferation and apoptosis of human tongue squamous cell carcinoma in vitro and in vivo
Wenyu GUO ; Lingping KONG ; Shanshan SUN ; Yu WANG ; Minghui ZHAO ; Xuan ZHOU ; Xudong WANG ; Lun ZHANG
Tianjin Medical Journal 2016;44(10):1185-1189
Objective To investigate the influence of long non-coding RNA HOTAIR in proliferation and apoptosis of human tongue squamous cell carcinoma in vitro and in vivo. Methods siHOTAIR was used to inhibit the HOTAIR expression in Tb3.1 human tongue squamous cell carcinoma cell line. The experiments were divided into siHOTAIR group, nonsense sequence group and blank control group. Real-time PCR was used to detect the HOTAIR expression. MTT assay was employed to determine the cell survival. The expression levels of Bcl2, BAX, caspase-3, cleaved caspase-3 were examined by Western blot assay. Tb3.1 xenograft tumor model was established in BALB/c nude mice, and the tumor model was divided into control group, negative group, and siHOTAIR treated group. The tumor tissues were measured by immunohistochemistry stain (IHC) and TUNEL assay. Results The detection of real-time PCR showed that HOTAIR expression was reduced after treated with siHOTAIR. Western blots assay showed that Bcl-2 protein was suppressed while cleaved caspase-3 and BAX protein were up-regulated after treated with siHOTAIR. MTT assay indicated that the cell survival rate was significantly reduced in siHOTAIR treated group. Flow cytometry detected that apoptosis levels were increased in siHOTAIR group. The level of cell senescence was higher in the siHOTAIR group than that of control group. Results of IHC indicated that Ki-67 and Bcl-2 protein of tumor tissue were inhibited, while BAX and cleaved caspase-3protein expressions were elevated simultaneously in the siHOTAIR group. TUNEL assay suggested that more apoptosis was observed in siHOTAIR group. Conclusion HOTAIR can affect proliferation and apoptosis of tongue squamous cancer cells. HOTAIR may be one of the new candidate targets for human tongue cancer therapy.
2.Diagnosis and prognosis of 280 patients with pancreatic carcinoma
Huanjing WANG ; Fachao ZHI ; Xinmei ZHAO ; Chaolan LV ; Weijian LUN ; Sanxi ZHOU ; Bo JIANG
Chinese Journal of Pancreatology 2010;10(1):2-5
Objective To evaluate the factors influencing early diagnosis and prognosis in patients with pancreatic carcinoma.Methods The clinical data of 280 patients who had complete follow-up data with pancreatic carcinoma treated from January 2002 to January 2007 were reviewed retrospectively.The medical history and follow-up data were collected from all patients.Survival rate was calculated by the life table method and the Kaplan-Meier estimation.Log-rank test was used for univariate prognostic analysis and Cox regression was used for multivariate prognostic analysis.Results 91.8%of the patients were more than 40 years old and the peak age was 50~73 years old;the major presentations were abdominal pain and jaundice.Major imaging tests included B-ultrasound and CT,the sensitivity was 70.6%,95.3%,respectively;89.3%of patients had combined B-ultrasound and CT examination.The sensitivity of CA19-9 was 81.1%.The median survival time was(7.0±0.5)months.Overall survival rates at 1~5 year survival rates were 28%,9%,6%,2%,and 1%.Univariate analysis suggested that age>65 years old,CA19-9>mean value,TNM Ⅲ or Ⅳ stage,lymph nodes invasion,vascular invasion,and metastasis of two or more organs,non-surgical treatment,KPS score<60 points,weight loss≥5 kg were poor prognostic factors;Cox multivariate analysis showed that treatment modalities,age,TNM stage,KPS score and ascites were independent risk factors for dismal prognosis.Conclusions The age,ascites,tumor stage and treatment modalities affected the prognosis of patients with pancreatic cancer.Early diagnosis and treatment was important to improve the survival time of patients with pancreatic cancer.
4.Effects of acupuncture at Shenmen (HT7) or Taiyuan (LU9) on P3a and P3b of event-related potentials in healthy young adults.
Ronglin CAI ; Ling HU ; Zijian WU ; Keming WANG ; Lu HE ; Yiping ZHOU ; Lun ZHAO ; Wubin HU
Journal of Integrative Medicine 2012;10(1):48-53
To investigate the effects of manual acupuncture at Shenmen (HT7) or Taiyuan (LU9) on the attention function of the brain, and to lay an experimental foundation for researching brain function and integration mechanisms of the human brain in relation to acupuncture stimulation.
5.Analysis of monitoring results of urinary iodine of children aged 8 - 10 of Zhangjiakou city in Hebei province, 2009
Zhong-xiang, WANG ; Pei-lun, HUA ; Jie, TAN ; Yu-min, ZHANG ; Yan-qing, ZHAO
Chinese Journal of Endemiology 2012;31(1):81-83
Objective To analyze the monitoring results of urinary iodine of students aged 8 - 10 in Zhangjiakou city,problems in monitoring results,and to provide basic information for working out control strategies of iodine deficiency disorders.Methods A township(town,street) in each country of each city(district) in Zhangjiakou was selected according to 5 positions of the east,the west,the south,the north and center,and 1 village elementary school was sampled in each chosen township,twenty students(half male and female) aged 8 - 10 were selected to collect their urine samples in each school.Urinary iodine concentration was determined by arseniccerium method.Results The median of urinary iodine of the 1700 children aged 8 - 10 was 291.5 μg/L,with < 50 μg/L accounted for 0.8%(13/1700),50 ~ 99 μg/L about 4.9%(83/1700),100 - 199 μg/L about 20.5% (349/1700),200 - 299 μg/L about 29.7%(504/1700),and ≥300 μg/L about 44.9%(764/1700).Conclusions Urinary iodine has reached the elimination standard of iodine deficiency disorders in Zhangjiakou city.But the situation of more than adequate amount of urinary iodine and iodine excess is relatively serious and it is necessary to lower iodine concentration.
6.Monitoring results of iodized salt of Zhangjiakou city in Hebei province during 2001-2009
Wen-jun, LI ; Yu-min, ZHANG ; Pei-lun, HUA ; Zong-xiang, WANG ; Yan-qing, ZHAO
Chinese Journal of Endemiology 2011;30(4):424-426
Objective To analyze comprehensively the monitoring data of iodized salt in Zhangjiakou city during 2001 to 2009, and to provide basic information for working out control strategies of the iodine deficiency disorders. Methods According to the iodized salt monitoring requirements in National Iodine Deficiency Disorders Monitoring Program of Ministry of Health, a batch of nine salt samples were taken from each processing (wholesale)company of each county or district of the seventeen counties(districts) of Zhangjiakou once a month. Two townships (towns, street offices) were selected by their location of east, south, west and north in each county(district), and a township in central area each year. Four villages(neighborhoods) were selected in each township(town, street office),and eight household salt samples were collected in each village(neighborhood), and quantitatively determined by direct titration of iodine. Results Iodized salt processing(wholesale) : during 2001 to 2009, a total of 1728 batches was monitored, 1689 batch qualified, batch qualification rate 97.74%;15552 salt samples were tested, 15 357 qualified, iodized salt qualification rate 98.75 %. Household salt levels : 5297 villages (neighborhoods) of 1305 townships(towns, street offices) were monitored, 44 316 salt samples were collected, 43 274 qualified, iodized salt qualification rate 98.04%(43 274/44 141 ), iodized salt coverage rate 99.61%(44 141/44 316), qualified iodized salt consumption rate 97.65%(43 274/44 316). Rate of non-iodized salt was 0.40%(260/44 316), and salt median iodine was 30.02 mg/kg. Conclusions The iodized salt quality indicators are within the state-controlled range in Zhangjiakou city for nine years which remaines at relatively stable levels with a smaller range of annual fluctuations.Detection of non-iodized salt over the years has become the main factors affecting the effectiveness of the prevention and control measures.We should increase monitoring,supervision,and universal health education,and prevent the spread of non-iodized salt.
7.CDK5 and epithelial-mesenchymal transition related proteins are abnormally expressed in head and neck squamous cell carcinoma
Minghui ZHAO ; Yuanyuan HUANG ; Shanshan SUN ; Lingping KONG ; Yu WANG ; Wenyu GUO ; Xuan ZHOU ; Xudong WANG ; Lun ZHANG
Tianjin Medical Journal 2015;(12):1408-1412
Objective To explore the expressions of Cyclin-dependent kinase 5 (CDK5) and Epithelial-Mesenchymal Transition (EMT) related proteins including N-cadherin, Vimentin and E-cadherin in head and neck squamous cell carcino? ma (HNSCC), and to determine the relationship between the expression of CDK5 and prognosis. Methods The expression levels of CDK5 and EMT related proteins were evaluated by immunohistochemistry in 55 patients who were diagnosed as HN?SCC. They were also analyzed in different clinical pathological factors. The correlation of CDK5 and EMT related proteins as well as the relationship between the expression of CDK5 and prognosis were also analyzed. Results The expression level of CDK5 was significantly higher in patients with lymph node metastasis than that in patients with non-lymph node metastasis (91.67%vs 30.23%, P<0.05). It’s also higher in T3-T4 stages than that in T1-T2 stages (85%vs 20%, P<0.05). The ex?pression levels of N-cadherin and Vimentin were significantly higher in patients with lymph node metastasis than those in patients with non-lymph node metastasis (75.00%vs 6.98%;91.67%vs 27.91%, all P<0.05). However, the expression level of E-cadherin was significantly lower in patients with lymph node metastasis (8.33%vs 86.05%, P<0.05) compared to that in patients without. CDK5 was positively correlated with N-cadherin and Vimentin, but negatively correlated with E-cad?herin (rs=0.512, 0.443,-0.363, all P<0.01). The 3-year survival rates were significantly lower in patients with high expres?sion of CDK5 (37.5%) than that in patients with low expression of CDK5 (87%, Log-rankχ2=12.678, P<0.01). Conclusion CDK5 and EMT related proteins were activated abnormally in HNSCC with lymph node metastasis. CDK5 may be a new bio?logical marker for prognosis of HNSCC.
8.A comparison study on the detection of anti-human immunodeficiency virus type 1 (HTV-1) antibodies in different populations with a new rapid test using oral mucosal transudate samples versus enzyme linked immunosorbent assay (ELISA) using serum samples
Yan WU ; Kerong WANG ; Jing HAN ; Hongxin ZHAO ; Hui ZENG ; Keyi XU ; Yanchun LIU ; Huiwen YAN ; Xingwang LI ; Wenhui LUN
Chinese Journal of Dermatology 2011;44(5):302-305
Objective To evaluate the consistence in the detection of antibodies against HIV-1 between a new rapid test using oral mucosal transudate (OMT) samples and ELISA using serum samples. Methods Two-hundred patients who were positive for anti-HIV-1 antibodies by serum ELISA and confirmed by Western blot to be infected with HIV, and 600 healthy human controls negative for anti-HIV-1 antibodies by serum ELISA, were eligible for this study. OMT samples were collected from these subjects and subjected to a rapid test for anti-HIV-1 antibodies. The factors influencing the performance of the rapid test were analyzed. Results Of the 200 OMT specimens from HIV-infected patients, 198 showed positive reaction, 2 showed negative reaction. Among the 198 positive reactions, 192 (96%) were "clear" and easy to make decisions, 4 (2%) were "faint", 2(1%) were "very faint" and required professionals to make decisions. The rapid test was negative in all the 600 OMT specimens from the control group. Conclusions The consistence in the detection of anti-HIV-1 antibodies between the OMT rapid test and serum ELISA was 99% in HIV-positive specimens, 100% in HIV-negative specimens, and 99.75% in all the specimens.
9.Influence of silencing TMPRSS4 expression on growth and invasion of pancreatic cancer SW1990 cell
Huanjing WANG ; Fachao ZHI ; Xinmei ZHAO ; Haitao QING ; Weijian LUN ; Sanxi ZHOU ; Wen GUO ; Tunming CHENG ; Bo JIANG
Chinese Journal of Pancreatology 2011;11(3):187-189
Objective To study the influence of the small interfering RNA (siRNA) interference TMPRSS4 expression on human pancreatic cancer SW1990 cell's proliferation and invasion. Methods The four eukaryotic expression vector of TMPRSS4 gene were synthesized in vitro and were transfected transiently into human pancreatic cancer SW1990 cells. TMPRSS4 mRNA expression of transfected cells was detected by real-time RT-PCR. The most efficient eukaryotic expression vector was used to be transfected into SW1990 cells. By using G418, cell strain that can silence TMPRSS4 gene stably was screened. The TMPRSS4 mRNA expression of the stable cell strain was detected by real time PCR TMPRSS4 protein expression was detected by western blot. The proliferation ability of transfected SW1990 cells was detected by CCK-8 method. By Transwell, the invasion change of SW1990 cell was detected. Results A stable cell strain, SW1990/psi TMPRSS4, was successfully constructed, in which the expression level of TMPRSS4 could be reduced stably by RNA interference. Cell transfection efficiency was 82.9%. Compared with the control group, the TMPRSS4 mRNA and protein levels were reduced by 80.1% and 60% ,and number of penetrating cells was 118.6 ±13.4 in SW1990/psi TMPRSS4 group, which was significantly lower than those in the negative control group (157.4 ± 12.9) and control group (157.0±9.5, P <0.01). Cells invasion inhibitory rate was 24.5% in SW1990/psi TMPRSS4 group. The cell proliferation was not significantly different among all the groups. Conclusions A stable cell strain is screened successfully in which the expression level of TMPRSS4 can be reduced stably. The down-regulation of TMPRSS4 gene expression level can inhibit the invasion of SW1990 cells, but has no effect on cell proliferation.
10.Expression of human gene 5 transactivated by pre-S1 protein of hepatitis B virus in yeast cell
Jian-Kang ZHANG ; Jun CHENG ; Jiang GUO ; Yong-Zhi LUN ; Dan-Qiong WANG ; Long-Feng ZHAO ; Yuan HONG ; Yu MAO ;
Chinese Journal of Infectious Diseases 2007;0(11):-
Objective To study the exact function of human gene 5 transactivated by pre-S1 protein of hepatitis B virus(PS1TP5)by investigating the gene expression of PS1TP5 in yeast cells. Methods Reverse transcription-polymerase chain reaction(RT-PCR)was performed to amplify the gene of PS1TP5 using the mRNA of HepG2 cells as template and the gene was cloned into pGEM-T vector.The gene of PS1TP5 was cut from pGEM-T-PS1TP5 vector and cloned into yeast expressive plasmid pGBKT7,then pGBKT7-PS1TP5 was transformed into yeast cell AH109.The yeast protein was isolated and analyzed with sodium dodecyl sulfate-polyacrylamide gel electrophoresis(SDS-PAGE) and Western hybridization.Results PS1TP5 gene was successfully amplified and identified by DNA sequencing.The digested fragment was cloned into pGBKT7 vector and transformed into yeast cell AH109.The results of SDS-PAGE and Western assay showed that the relative molecular weight of the expressed product was about 36 950,and PS1TP5 protein existed in yeast cells.Conclusion The findings suggest that PS1TP5 can be successfully expressed in yeast cell.