1.Research progress on HOTAIR in tumor biological behaviors
Xuan ZHOU ; Lingping KONG ; Lun ZHANG
Chinese Journal of Clinical Oncology 2016;43(16):731-734
Long non-coding RNA (lncRNA) is a cluster with over 200 bp in length and has no protein-encoding product RNA, which is in-volved in cellular physiological or pathological process, especially in human oncogenesis. HOX transcript antisense RNA (HOTAIR), which is 2158 bp in length, is one of the most well-studied lncRNAs. Overexpression of HOTAIR is correlated with oncogenesis or me-tastasis in numerous epithelium original human cancers, including breast and colorectal cancers. Inhibiting HOTAIR expression could suppress cell growth and invasive ability of tumors. This review provides a brief summary of the latest progress in lncRNA-based can-cer research.
2.Clinical Study on the Combination of Western Medicine with Acupuncture for Treatment of Polycystic Ovary Syndrome of Infertility
Xuan SUN ; Qihua QI ; Shoufang KONG ; Jinyan ZHONG
Chinese Journal of Information on Traditional Chinese Medicine 2016;23(11):25-28
Objective To observe the clinical efficacy of the combination of western medicine with acupuncture for treatment of polycystic ovary syndrome (PCOS) of infertility.Methods Totally 120 PCOS patients were randomly divided into western medicine group and acupuncture group, 60 cases in each group. All patients took Diane-35, and with whom resistant to insulin were added with Metformin. At the meantime patients increased the amount of exercise with adjustment and control of everyday diet. After three periods, the western medicine group used the method of Clomifene Citrate with Chorionic Gonadotropin to help ovulation induction. The acupuncture group used the method of acupoint selection including invigorating spleen and kidney with adjusting qi and blood to promote ovulation. The level of FSH, LH, FINS, E2 and T of the two groups were observed before and after treatment.Results After treatment, the levels of T, FINS, LH and BMI in the two groups decreased, with statistical significance (P<0.05). The levels of T and BMI in the acupuncture group were lower than the western medicine group after the treatment, with statistical significance (P<0.05). The total efficiency, gestation number, number of high-quality follicle, and basal body temperature of acupuncture group were all higher than the western medicine group, with statistical significance (P<0.05).Conclusion The combination of western medicine with acupuncture for treatment of PCOS of infertility has confirmed efficacy.
4.Expression of Fos positive neurons in vomiting reflex regions of brain in non-vomiting rats after intraperitoneal injection of emetic
Zhe KONG ; Shumei JI ; Xuan GAO ; Yingcheng CHEN ; Zhenlong GUAN
Chinese Journal of Tissue Engineering Research 2005;9(21):222-224
BACKGROUND: It is discovered by administrating different emetics to vomiting animals, like cats, that there are a large amount of Fos positive neuronal expressions in the arc region from nucleus of solitary tract, lateral tegmentum to ventrolateral area. And it has been viewed that the arc region from area postrema, nucleus of solitary tract to ventrolateral reticular structure is the main emetic region. Whether do the non-vomiting animals reflect in response or not after emetic injection?OBJECTIVE: To observe the distribution of Fos positive neurons in relevant emetic regions of brain and spinal cord in rats after abdominal injection of emetic, cisplatin.DESIGN: Randomized controlled experiment based on animals.SETTING: Neural Physiological Research Room of Life Science College in Hebei Normal University and Physiological Room of Basic Medicine Institute in Hebei Medical University.MATERIALS: The experiment was performed in Neural Physiological Research Room of Life Science College in Hebei Normal University and Physiological Room of Basic Medicine Institute in Hebei Medical University from March to August 2003. Twelve SD male rats were employed, body weighted varied from 220 to 250 g, of clean-grade. They were randomized into experimental group of 6 rats and the control of 6 rats.INTERVENTIONS: In experimental group, the emetic, cisplatin, was injected abdominally 10 mg/kg. In the control, the physiological saline of same dose was injected. Afterwards, the activity changes in rats were observed at room temperature, quiet and light-avoided environment. Six hours later, the brain tissue was collected for frozen continuous sectioning. Immunohistochemical staining method was used to observe the distribution of Fos positive neurons in brainstem and forebrain nuclei and to count positive cell.MAIN OUTCOME MEASURES: ① Behavior observation in rats after emetic injection. ② Counts of Fos positive cell in relevant regions of brain in rats.RESULTS: Twelve rats all entered result analysis. ① In 20 minutes after injection, the rats in both groups were in tranquilizing state, lying prone with body curled, almost without any movement. In 60 minutes after injection, the rats in the control were recovered to normal, free of eating or drinking. The rats in experimental group were in prone-lying state with body curled. They rose up or shook the heads occasionally, and they breathed fast and uneven.In 2 hours after injection, in experimental group, the rats were still in abdominal prone tightly in the cage, with heads lowed and irregular shaking of noses. In 5 hours, the rats in experimental group began standing up and moving, with normal breathing, but they still did not eat or drink. ② Fos positive neurons in solitary tract, area postrema and lateral parabrachial nucleus and paraventricular nucleus, supraoptic nucleus and arc nucleus in hypothalamus (64.3 ±9.6, 83.4 ±15.0, 148.8 ±19.9, 80. 2 ± 11.8, 20.7 ±3.8, 86. 6 ± 10.8) were remarkably higher than those in the control(56. 2 ±6.3,73.5±9.9,136.9±17.8,66. 1±10.3,17.3±3.4,78.8±10.5).CONCLUSION: Emetics induce discomforts in internal organs of rats, due to which, there probably exist emetic regions similar to vomiting animals in central neural system. But it is probably lack of vomiting-related adjusting mechanism. Emetics irritate the increase of Fos positive neurons in relevant regions in the brain of rat, which suggests that there exist relevant neural chemical pathways similar to nausea in the brain of non-vomiting rats.
5.Clinical value of thinprep cytology test combined with colposcopy in diagnosis of cervical lesion
Lili KONG ; Lijing XUAN ; Yanying LI ; Limin ZHAO
Clinical Medicine of China 2010;26(6):637-639
Objective To investigate the clinical value of thinprep cytology test(TCT) combined with colposcopy in diagnosis of cervical lesion. Methods Retrospective analysis was conducted in 174 females with abnormal cervical smear. All the patients underwent colposcopy to screen the cervical lesions. According to the pathological results,the clinical value of TCT and colposcopy was analyzed and compared. Results In the 174 cases with abnormal cervical smear,the positive rate of atypical squamous cells(ASC) was 27. 0% (47/174). There were 92 low grade squamous intraepithelial lesion (LSIL) (52. 9%) ,33 high grade squamous intraepithelial lesion (HSIL) (19. 0%) and 2 squamous cell cancinoma (SCC) (1. 1%) . According to the diagnosis by biopsy and pathology,the positive rate of benign cervical cell(BCC) was 16. 1% (28/174) .cervical intraepithelial neoplnsia (CIN) I 42. 0% (73/174), CIN II 18.4% (32/174), CIN ffl 21. 8% (38/174) and SCC 1. 7% (3/174) . The coincidence rate of TCT and pathology was 83. 9% (146/174). Conclusions TCT and colposcopy are a practical approach for detecting cervical lesion. TCT combined with colposcopy is the optimal screening approach for cervical lesion, which can enhance the sensitivity and specificity of the diagnosis of cervical lesions.
6.Expression and significance of miRNA-34a in cervical intraepithelial neoplasia
Hui YUAN ; Shoufang KONG ; Jinyan ZHONG ; Xuan SUN
Chinese Journal of Primary Medicine and Pharmacy 2016;23(5):768-770
Objective To detect the expression of miRNA -34a in normal cervical,low grade cervical intra-epithelial neoplasia and high grade cervical intraepithelial neoplasia,to explore its clinical significance.Methods The expression levels of miRNA -34a in 82 cases of low grade cervical intraepithelial neoplasia (CINⅠ,CIN L group),87 cases of high grade cervical intraepithelial neoplasia (CINⅡ -Ⅲ,CIN H group) and 60 cases of normal cervical tissues (control group) in tissue specimens and serum were examined by real -time PCR(RT -PCR). Results The relative expression levels of miRNA -34a in tissues and serum of CIN H group were (0.584 ± 0.288),(0.528 ±0.299),which of CIN L group were (0.903 ±0.252),(0.847 ±0.279),which of the control group were (0.960 ±0.212),(0.908 ±0.223).The miRNA -34a level of CIN H group in tissues and serum were lower than CIN L group and control group,there was significant difference( Ptissues =0.001,Pserum =0.003).The miRNA -34a level in tissue and serum had no significant difference between the CINL group and control group(P >0.05).The expression level of serum miRNA -34a was positively related with the level of the tissues(r =0.907,P <0.01).Conclusion The expression level of miRNA -34a in high grade CIN was significantly decreased,which indi-cated that miRNA -34a play an important role in the prognosis of CIN and the transformation of cervical cancer,and it has certain clinical value for the diagnosis and treatment of CIN.
8.Construction and expression of procaryotic expression vector of a chimeric GPI-B7-1 cDNA
Jianwei WANG ; Yingbin LIU ; Yanjing CAO ; Xuan WANG ; Jiangtao LI ; Ying KONG ; Xiaoming MA ; Shuyou PENG
Chinese Journal of General Surgery 2009;24(1):38-41
Objective Through constructing prokaryotic expression vector pET-30a-GPI-B7-1, to gain purified GPI-B7-1 fusion protein so as to confirm the tumor immune effect. Methods The DNA fragment encoding the signal for GPI-anchor attachment of hPLAP-1 and the cDNA encoding the human costimulatory molecule CD80 ( BT-1 ) were cloned from fresh placenta and human peripheral blood monocytes (PBMC) respectively. The two fragment were annealed to form a fusion gene (GPI-BT-1) by PCR. Then the fusion gene was inserted into the prokaryotic expression vector pET-30a, resulting in pET-30a-GPI-BT-1. Transfer to E. coli BL21, purified fusion protein were analysed by SDS-PAGE and Western blot. Results Agarose gel electrophoresis map of GPI and BT-1 PCR products show that GPI goal gene strap was seen at 133bp region and BT-1 goal gene strap at 792 region. Identification of recombinant pET-30a-GPI-B7-1 by restriction enzyme and PCR illustrate two goal fragment for 5000 bp and 900 bp, to realize the expression of fusion gene ( GPI-B7-1 ) at the E. coli BL21. The fusion protein was successfully produced in the pET expression system induced by IPTG and purified by Ni2 + -NTA agarose column. By SDS-PAGE and Western blot analysis, the observed molecular weight of the fusion protein was 38 kDa. Conclusion The purified GPI-B7-1 fusion protein can be obtained from E. coli BL21 transfered by prokaryotic expression vector pET-30a-GPI-B7-1, which will prove useful tool for the study of tumor immune therapy.
9.The association between plasma Vitamin D3 and B cell subgroups in patients with primary Sj(o)gren's syndrome
Feifei KONG ; Li JIN ; Xiaomei LI ; Xuan FANG ; Xiangpei LI ; Guosheng WANG ; Long QIAN
Chinese Journal of Rheumatology 2014;18(3):151-154
Objective To investigate the plasma vitamin D3 level and its association with B cell subsets of patients with primary Sj(o)gren's syndrome (pSS).The role of vitamin D3 levels in the pathogenesis of pSS was explored.Methods The expression of plasma vitamin D3 levels of 55 patients with pSS and 32 controls were analyzed by ELISA.Frequencies of peripheral blood CD19+CD27-na(i)ve B cells,CD19+CD27+ memory B cells and CD19+CD27high plasma cells were analyzed by flow cytometry in 34 pSS patients without therapy and 22 controls.The relationship between the vitamin D3 levels and B cell subsets,SSDAI,tear flow rate,saliva flow rate,rheumatoid factor,immunoglobulin was analyzed in pSS patients.Non-parametric test,t test,one-way ANOVA,x2test,Pearman's and Spearman's correlation analysis were used for statistical analysis.Results ① There was significant difference in the levels of plasma 1,25 (OH)2D3 between the pSS patients group and normal control group,1,25 (OH)2D3 was significantly lower in pSS patients than that in the normal control group [24.17(22.20,28.41) pg/ml and 41.25(23.38,62.18) pg/ml,P<0.05],and that was also obviously lower in the active group [22.64(20.74,24.90) pg/ml] than that in the normal control group (P<0.05),and that was also obviously lower in the active disease group than that in the inactive disease group [25.39 (23.16,33.09) pg/ml,P<0.05],but there was no difference between the inactive group and the normal control group (P>0.05).② The percentage of peripheral blood of CD19+CD27high plasma cells and CD19+CD27+ memory B cells in CD19+ cells was reduced in patients in the pSS group compared with the control group [(0.89±0.30)% and (1.72±0.43)%,(24±8)% and (34±5)%; P<0.05],and that was also significantly lower in the active group [(1.03±0.59) % ; (26± 10)%] and inactive group [(1.00±0.16)%,(26± 3)%] than that in the normal controls (P<0.05).However,there was no difference between the active group and the inactive group (P>0.05),but the frequency of peripheral blood of CD19+ CD27-naive B cells in CD19+ B cells was increased in patients with pSS compared with normal control group [(75.4±7.5)% and (63.9±5.2)%,P<0.05],and that was also significantly higher in the active group [(73.4±9.7)%] and inactive group [(73.3±2.9)%] than that in the normal control (P<0.05),there was no difference between the active group and the inactive group.③ Significant negative correlation was observed between 1,25 (OH)2D3 and the percentage of peripheral blood CD19+CD27+ memory B cells in CD19+ cells as well as immunoglobulin G(r=-0.627,P=0.039; r=-0.657,P<0.01) level.Conclusions These results demonstrate that abnormality of vitamin D levels may play an important role in the pathogenesis of pSS.
10.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.